Chikungunya virus infection can cause chikungunya hemorrhagic fever, which is prevalent in a wide range of tropical and temperate regions and has become a global public health problem. Chikungunya hemorrhagic fever is mainly treated by symptomatic treatment, and there is no specific antiviral drug. Vaccination is an economical and effective means to prevent chikungunya virus infection. Although two vaccines have been approved for use in local areas, they are far from meeting the demand for vaccines in high-risk areas. This article reviews the research and development status of chikungunya virus vaccine, focusing on vaccines on the market, candidate vaccines entering clinical trials and vaccines that have potential to enter the clinical trial.

Objective To establish and validate the detection method of coxsackievirus A6 (CV-A6) neutralizing antibody by micro-cytopathic effect (CPE) method.Methods A neutralizing antibody detection method based on CPE was established using human rhabdomyosarcoma (RD) cells and CV-A6 standard detection strains. It was verified for relative accuracy, precision, linearity, specificity, and durability.Results A determination method of neutralizing antibody titer for CV-A6 immune serum was established. Three serum reference samples of different dilutions (1×, 16×, and 256×) were measured 3 times with relative biases of ﹣2%, ﹣4%, and ﹣16%, respectively. The slope of the fitted linear regression was 1.065, indicating good relative accuracy. The geometric coefficients of variation (GCVs) were 19%-50%, and the repeatability was good. The GCVs of 10 mouse immune sera neutralizing antibody titers measured for 3 times on different days were 0%-75%, indicating good intermediate precision. The regression coefficient of best fit line for the neutralizing antibody titers of CV-A6 neutralizing antibody serum reference at 9 dilution levels measured 3 times was 0.93, and the linear regression was statistically significant (F=340.99, P<0.000 1). The neutralizing titers of negative sera were all less than 8, showing good specificity. The GCVs of different RD cell passages, different RD cell densities, different viral loads, different culture days, different number of repeated freeze-thaw times and short-term storage time range of serum to be tested were 23%-101%, indicating good durability.Conclusions The CV-A6 neutralizing antibody detection method has been successfully established with good relative accuracy, precision, linearity, specificity, and durability. It can be used to evaluate the neutralizing antibody levels of immune sera against CV-A6.

Chikungunya virus (CHIKV) is a type of alphavirus transmitted by Aedes mosquitoes. CHIKV infection can result in chikungunya fever, which presents with symptoms such as high fever, joint pain, and chronic arthritis, among other clinical manifestations.In recent years, it has caused multiple outbreaks globally, leading to serious public health issues.However, there are currently no specific antiviral drugs approved for marketing, and clinical treatments mainly focus on symptomatic and supportive treatments. With the expanding geographic range of CHIKV transmission and increasing disease burden, there is an urgent need to develop effective antiviral drugs. This article reviews the epidemiology, genomic structure, and function of CHIKV, as well as recent research progress on antiviral drugs against CHIKV, including direct-acting antiviral agents and host-targeted drugs. Additionally, this article discusses the challenges faced during the drug development process, such as the occurrence of drug-resistant mutations, drug safety, and clinical translation issues. It also looks forward to future research directions, including multi-target drug design, combination therapy strategies, and computer-aided drug design with artificial intelligence. By systematically summarizing existing research findings, this article aims to provide guidance and insights for the further development of antiviral drugs against CHIKV.

Objective To establish and validate a method for the simultaneous determination of residual Triton X-100 and polysorbate 80 (PS80) in influenza virus split vaccine (MDCK cell).Methods Triton X-100 and PS80 residues in influenza virus split vaccine were detected by high performance liquid chromatography (HPLC)-evaporative light-scattering detector (ELSD) method. The specificity, accuracy, precision, linearity, limit of quantitation and robustness of this method were verified.Results In the HPLC-ELSD analysis, no chromatographic peaks were observed in blank solvent. Two target chromatographic peaks were visible in both mixed standard solution and test sample, with the resolution from adjacent peaks greater than 1.5. The spiked recovery rates of Triton X-100 and PS80 at various concentrations were 90.0%-105.0% and 80.0%-105.0%, respectively. When the same experimenter performed repeated injections six times, the relative standard deviations (RSDs) of Triton X-100 and PS80 contents were 0.7% and 1.4%, respectively. When two inspectors determined the residual amounts of Triton X-100 and PS80 on different working days, the RSDs were both ≤ 5.0%. When Triton X-100 mass fraction was in the range of 0.010%-0.100%, a good linear relationship was observed between the concentration and peak area, with limit of quantitation at 0.010%. When PS80 mass fraction was in the range of 0.006%-0.060%, a linear relationship was found between the concentration and peak area, and the limit of quantitation was 0.003%.Conclusions HPLC-ELSD detection method for Triton X-100 and PS80 residues in influenza virus split vaccine is successfully established. The specificity, accuracy, precision, linearity and robustness are good.

Objective To develop a purification process for anti-receptor activator of NF-κB ligand (RANKL) monoclonal antibodies.Methods The upstream cell culture supernatant was clarified by depth filtration to remove cells. The clarified harvest was then subjected to affinity chromatography (AC) capture, low-pH viral inactivation, anion-exchange chromatography (AEX), cation-exchange chromatography (CEX), viral filtration (VF), ultrafiltration/diafiltration (UF/DF), and sterile filtration to obtain the antibody bulk. AC purification efficiency was evaluated based on the yield and removal of product-related impurities. The dynamic binding capacity (DBC) of resin was calculated by monitoring the breakthrough point. Low-pH viral inactivation conditions were determined by evaluating the impact of pH and temperature conditions on sample purity and activity.The optimal AEX buffer system was determined by assessing the stability of target protein in different buffer conditions and the removal of process-related impurities. Purification efficiency and DBC were also evaluated. CEX process parameters were optimized using linear gradient elution and fraction collection to analyze impurity clearance. The VF filter type was selected based on throughput, flux, and flow decay.Membrane pore size, flow rate, and transmembrane pressure were determined based on UF/DF and sterile filtration manufacturer specifications and platform technology. Concentration and dialysis buffer exchange were performed accordingly. Finally, the bulk was obtained through a 0.2 μm membrane sterile filtration, and the purity and impurity were accessed.Results AC used MabSelect SuRe resin (DBC: 45.3 mg/mL resin) with 25 mmol/L Tris-HCl (pH7.5) for equilibration and 150 mmol/L HAc (pH2.8) for elution. Viral inactivation was accomplished with pH3.5 for 1 h. AEX used Sepharose Q FF resin (max DBC: 60 mg/mL resin) with 50 mmol/L Tris-HAc (pH7.5) for equilibration. CEX used SP HP resin of 21.9 mg/mL loading capacity with linear elution from 100% buffer A (10 mmol/L citrate， pH5.5) to 100% buffer B (10 mmol/L citrate + 1 mol/L NaCl， pH5.5) over 20 column volumes. Viresolve Pro was used for VF and PLCTK 30 kDa ultrafiltration cassette (C-channel) was used in UF/DF to concentrate to (40.00 ± 5.00) mg/mL, followed by 10× diafiltration and sterile filtration to yield bulk. Both the purity and impurity levels met predefined standards.Conclusion A robust process for anti-RANKL monoclonal antibody purification is successfully developed, yielding drug substance with acceptable qualities.

Objective To evaluate the optimal mouse immunogenicity model of 13-valent pneumococcal polysaccharide conjugate vaccine and the optimal mouse gender and immune dose by comparing the immune response in BALB/c, KM, and CD1 mice.Methods The 13-valent pneumococcal polysaccharide conjugate vaccine was used to immunize 3 strains of mice with doses of 0.05, 0.10, 0.20 and 0.40 μg, respectively. ELISA was used to analyze the immune responses to evaluate the optimal mouse model. Response of male and female mice of the optimal model immunized with 4 different doses was compared to evaluate the optimal mouse gender. Response of optimal mouse model of optimal gender immunized with 4 different doses was compared to evaluate the optimal immune dose.Results The immune response values of most serotypes were the highest in CD1 mice, and the weakest in KM mice. There was a significant dose-response relationship in CD1 mice. Immune responses of both male and female mice were low at 0.05 μg dose. At doses of 0.10, 0.20, and 0.40 μg, almost all serotypes showed higher immune responses in female mice than in male mice. The immune response of CD1 female mice immunized with 0.20 μg was the highset in most serotypes except 3, 9V, 19F, and 23F.Conclusion CD1 female mouse is the optimal immunogenicity evaluation model for 13-valent pneumococcal polysaccharide conjugate vaccine, and 0.20 μg is the optimal immunization dose.

Objective To provide basis for the quality control of impurities in oral hexavalent reassortant live attenuated rotavirus vaccine (Vero cell) by analyzing the residual levels of porcine trypsin, bovine serum albumin（BSA）, Vero cell DNA，and the DNA fragment size in monovalent virus bulk.Methods The residual levels of porcine trypsin and BSA in 30 batches of monovalent virus bulk were detected by ELISA. The residual Vero cell DNA in 30 batches of monovalent virus bulk was purified by magnetic bead extraction method and then quantified by fluorescent staining method, and the size and distribution of Vero cell DNA fragment were detected by capillary gel electrophoresis.Results In 30 batches of monovalent virus bulk tested, the residual levels of porcine trypsin was 3.8-7.2 μg/mL, BSA was 85-268 ng/mL, and Vero cell DNA was 48-115 μg/mL. The DNA fragments of Vero cells in the range of 201- 2 000 bp counted for 73.4%-91.9% of fragments.Conclusion The residual impurities in the monovalent virus bulk of oral hexavalent reassortant live attenuated rotavirus vaccine (Vero cell) are controllable.

Objective To analyze pathogens, severe cases and population distribution characteristics of hand foot mouth disease (HFMD) after the launch of inactivated enterovirus A71 (EV-A71) vaccine in China, and to provide references for the prevention and control of HFMD in China.Methods The literature published openly from 2016 to 2025, explicitly involving the application effect evaluation of EV-A71 vaccine and containing basic data such as the changes of pathogens before and after the application of vaccine, severe cases or incidence rates, and population distribution characteristics, was collected from China National Knowledge Infrastructure and comprehensively analyzed by chi-square test.Results The population distribution of HFMD cases before and after the use of EV-A71 inactivated vaccine showed that scattered children accounted for the highest proportion, followed by children in kindergartens and nurseries, and students and other groups accounted for the lowest proportion. However, after the application of EV-A71 vaccine, the composition of scattered children in HFMD cases decreased by 11.35% compared with that before the vaccine application, the proportion of children in kindergartens and nurseries increased by 25.62%, and the proportion of students and others increased by 61.38%. The differences in population distribution before and after the vaccine application were all statistically significant (χ²=15 659.45, 11 028.25, 3 587.36; P<0.001). After the application of EV-A71 vaccine, the average number of HFMD cases caused by EV-A71 decreased by 56.17%, while the average number of cases caused by coxsackie virus A16 (CVA16) and other enteroviruses increased by 8.08% and 62.37%, respectively, and the differences were all statistically significant (χ²=41.94, 310.80, 39.37; P<0.001). Non-EV-A71 enteroviruses (such as CVA16, CVA6 and CVA10, etc.) became dominant pathogens. After the application of vaccine, the average number of severe cases decreased by 42.70%, and the proportion of those caused by EV-A71 in severe cases dropped by 17.89%. The differences were statistically significant (χ²=4 087.32, 4 215.92；P< 0.001). However, approximately 59% of severe cases were still caused by EV-A71 infection after the use of the vaccine, suggesting that EV-A71 remained the main pathogen leading to critical HFMD.Conclusion EV-A71 vaccine has significantly reduced the incidence of severe HFMD cases, but EV-A71 remains the primary pathogen causing severe cases, while non-EV-A71 enteroviruses have become dominant pathogens.

Objective To establish a capillary zone electrophoresis (CZE) method to detect the polysaccharide content and molecular size distribution of group A, C, Y, and W135 meningococcal polysaccharide vaccine.Methods CZE method was developed to detect all 4 polysaccharides, and the optimal separation voltage and temperature were investigated. Four polysaccharides were characterized in a single experiment, and their contents and molecular size distributions were determined. The method’s linearity, repeatability, accuracy, and specificity were validated.Results When contents of all 4 polysaccharides ranged from 0.081 3 to 0.487 5 μg/μL, the CZE method demonstrated good linearity with coefficient of determination > 0.98. For low, medium, and high concentration samples, the recovery rates were between 95% and 110%, and the relative standard deviation for 6 experiments was < 2.0%. No impurity peaks were observed at the target peak positions.Conclusion The CZE method shows good linearity, accuracy, repeatability, and specificity for detecting the four meningococcal polysaccharide, and is suitable for determining the polysaccharide content and molecular size distribution of group A, C, Y, and W135 meningococcal polysaccharide vaccine.

Objective To analyze the blocking effect of measures for preventing maternal-infant transmission of hepatitis B virus (HBV) in Qingdao and to explore related influencing factors.Methods A total of 869 HBV infected parturients giving birth in Qingdao in 2021 and their children who completed hepatitis B serological marker detection (HBV-exposed children) were selected as research objects. Through follow-up results and questionnaire surveys, the maternal-infant transmission rate of HBV and parents’ awareness of knowledge related to HBV maternal-infant transmission were obtained. Univariate and multivariate logistic regression methods were used to analyze the influencing factors of blocking failure.Results Of 869 HBV-exposed children surveyed, the maternal-infant transmission rate of HBV was 0.69%. Uncompleted full-course hepatitis B vaccination in HBV-exposed children [odds ratio (OR) = 0.030, 95% confidence interval (CI) = 0.004-0.232] and maternal premature rupture of fetal membrane(OR = 9.570, 95% CI: 1.343-68.201) might be independent risk factors for maternal-infant transmission of HBV. There were differences between parents of children in HBV surface antigen study group and control group in the awareness regarding necessary medication during pregnancy reducing the risk of maternal-infant transmission, the need for testing after HBV-exposed children receiving full-course hepatitis B vaccination, hepatitis B vaccination procedure, the number of vaccinations, and the intervention and blocking measures for HBV-exposed children after birth.Conclusions Enhancing prenatal care, reducing the occurrence of premature rupture of fetal membrane, and timely implementation of combined immunization strategies and full-course hepatitis B vaccination procedures for HBV-exposed children after birth can significantly improve the efficacy of maternal-infant blocking. Strengthening health education for key populations and improving the awareness rate of knowledge related to hepatitis B prevention are effective ways to reduce maternal-infant transmission of HBV.

Objective To evaluate the pharmacokinetic parameters of human coagulation factor Ⅸ (FⅨ) after single intravenous injection in patients with hemophilia B, and to preliminary analyze the clinical implications. Methods In a single-arm, open-label, multicenter clinical trial， 13 male patients aged 25-64 years with hemophilia B received a single intravenous injection of FⅨ at 50 international unit (IU)/kg under non hemorrhagic conditions. Blood samples were collected at 2 h before injection, 15 min, 30 min, and 1, 3, 9, 24, 48, 72, 96 h post-injection to measure FⅨ activity. The FⅨ pharmacokinetic parameters were calculated by non-compartment model. In addition, FⅨ inhibitors in patients were measured 30,90 d after injection.Results The plasma FⅨ concentrations peaked at 51.5-69.3 IU/dL in all patients after single intravenous injection of FⅨ for 15 min-1 h. The baseline-corrected pharmacokinetic parameters （±s） of FⅨ were below： peak time (0.37±0.22) h, elimination half-life (29.732±4.576) h, peak concentration (58.3±5.3) IU/dL, clearance (0.028±0.006) dL/kgh, area under the plasma drug concentration time curve (zero time to last time point) (1 626.189±285.365) %h, and incremental recovery rate (1.166±0.106) IUdL^-1/IUkg^-1. At 1 h post-injection, FⅨ activities were 40%-60% in all 13 patients, which maintained 40%-60% in 12 patients 3 h post-injection, and were still 40%-60% in 8 patients 9 h post-injection. No FⅨ inhibitors were detected in all 13 patients. Conclusion After single intravenous injection of FⅨ at 50 IU/kg in patients with hemophilia B, the major pharmacokinetic parameters, such as elimination half-life and clearance, indicate effective maintainence of blood concentrations in vivo, which can be used for further clinical studies on the efficacy of replacement therapy.

Objective To evaluate the non-clinical safety of live attenuated Japanese encephalitis vaccine with changed stabilizer by long term toxicity test in SD rats.Methods A total of 160 SD rats were subcutaneously injected with Japanese encephalitis vaccine with changed or unchanged stabilizer or NaCl(negative control) for 3 times. Animals were clinically observed, monitored for body weight, food intake, blood cell count, blood biochemistry, urinalysis, body temperature, clinical pathology, T lymphocyte subsets, and specific IgG antibody as well as performed urinalysis and ophthalmic examination.Results During the test, no significant abnormal reactions related to administration was observed. The body weight gain, food intake, blood cell count, and blood biochemistry showed statistically significant differences between certain vaccine groups and negative control （t=2.03-4.26，P=0.011-0.042），but no abnormal changes related to the stabilizer change were seen. There were no statistically significant changes in body temperature, coagulation function,and T lymphocyte subsets (t=1.35-1.98，P =0.052-0.186). By the end of recovery period, antibodies were detected in all animals in all vaccination groups, and there was no significant decrease in antibody titers. At 3 days after the last dose of administration and at the end of recovery period, no significant abnormal change was observed in the gross anatomy and histopathological examination of euthanized animals in all vaccination groups and negative control groups. There was no significant difference in toxicity, local irritation, and immunogenicity between vaccine groups with changed or unchanged stabilizer.Conclusion The long-term toxicity test results of live attenuated Japanese encephalitis vaccine with changed stabilizer in rats meet the requirements of animal safety evaluation.

Diabetes mellitus type 2 is a systemic endocrine metabolic disease characterized by insulin resistance and impaired function of pancreatic β cells. At present, most patients with diabetes mellitus type 2 are obese patients with metabolic disorders. Semaglutide is a glucagon-like peptide-1 receptor agonist， which can achieve blood glucose control and weight loss through multiple mechanisms, such as suppressing appetite, delaying gastric emptying, and improving pancreatic function. Semaglutide is widely used in the treatment of diabetes mellitus type 2. This article reviews the mechanism of action, research progress, clinical application issues and related suggestions of semaglutide in the treatment of diabetes mellitus type 2, aiming to provide a reference for the clinical application of semaglutide in the treatment of diabetes mellitus type 2 patients.

Bone injury is a common clinical disease, and existing clinical treatments are ineffective in treating some difficult conditions. The research results of bone tissue engineering provide a new solution to this problem. One mature bone tissue engineering injury repair method currently in use is bone marrow mesenchymal stem cell transplantation. However, there are several problems in its clinical application, such as the difficulty of seed cell sources, the lack of allogeneic stem cell seed cells or universal seed cells, etc. Recently, peripheral blood monocytes are found to have not only the potential for multidirectional differentiation, but also a strong proliferative capacity and availablility, thus can be used as an alternative material for bone injury repair. This paper reviews monocytes and their roles in bone injury repair and their mechanism, aiming at improving the life quality of patients with bone injuries and providing more effective treatment without side effects.

Objective To establish and validate a method for determination of total protein content after desorption in 13-valent pneumococcal conjugate vaccine.Methods The total protein content of the 13-valent pneumococcal conjugate vaccine was determined by the Lowry method after desorption. The linearity, accuracy, repeatability and durability of the established method were verified.Results The optimum desorption condition was to add 10% volume of 0.3 mol/L NaOH. Within the range of 40-200 μg/mL of protein standard, the linearity of the standard curve was good, coefficient of determination＞0.995. In the accuracy test, the recovery rates of protein standards at concentrations of 20,40,60,80 and 120 μg/mL ranged from 95.25% to 104.77%,demonstrating good accuracy. In the repeatability test,the relative standard deviation (RSD) of 6 test results of the sample at different time points was 3.82%, indicating good repeatability.In the durability test, no statistically significant difference (t = 0.059 and 0.238, P > 0.05) was observed in test results after samples were stored at room temperature for 15 or 30 min post-desorption, with RSD values of 4.82% and 5.14%，respectively, indicating good durability.Conclusion The established method can effectively, accurately and stably detect the total protein content of 13-valent pneumococcal conjugate vaccine.

Ankylosing spondylitis (AS) is a chronic autoimmune disease that primarily affects the sacroiliac joints, spine and adjacent tendons, soft tissues. The typical pathological progression of AS involves processes such as immune inflammation-driven responses, bone destruction, and new bone formation. These processes recur at same sites, ultimately leading to gradual joint stiffness and deformation. This article discusses the pathogenic mechanism， regulatory role, and current research status of IL-17 in AS, aiming to clarify the advantages of IL-17 as target in AS treatment and lay foundation for the development of IL-17-related drugs and the clinical management of AS.

In recent years， bispecific antibodies （BsAbs） have presented unique therapeutic potential in tumor immunotherapy by improving the targeted killing capacity of immune cells and blocking multiple signaling pathways to produce synergistic effects. Over 200 BsAbs for antitumor therapy are currently in clinical development worldwide， with 15 approved for use in hematologic malignancies or solid tumors. BsAbs are effective in treating malignant hematologic tumors. However， many challenges remain in solid tumor therapy， including target selection， drug delivery， safety and drug resistance. This article summarizes the clinical applications and research progress of licensed BsAbs in cancer treatment， providing a reference for BsAbs development and clinical utilization.

Objective To establish, verify and preliminarily apply a real time fluorescent quantitative reverse transcription PCR (RT-qPCR) detection method for xenotropic murine leukemia virus (X-MuLV). Methods Specific primers were designed according to gene sequence of X-MuLV provided by American Type Culture Collection. Recombinant plasmid and RNA standard containing target sequence were constructed, quantified through digital PCR and serially diluted to establish RT-qPCR method. Sensitivity, specificity, accuracy, precision, and robustness of RT-qPCR were investigated. The method was applied to validate virus clearance procedure by nanofiltration and chromatography, and compared with cytopathic effect assay.Results The established RT-qPCR detection method for X-MuLV showed a linear range of 2.5×10¹-2.5×10⁸ copies/μL，and the amplification efficiency was 98%. The sensitivity of detection was 2.5×10¹ copies /μL. No cross-reaction with other commonly used viruses in the lab was detected. The coefficients of variation of repeatability and intermediate precision validation were all less than 1%. Protein and buffer had no matrix effect on virus detection of samples. When the method was employed to access the X-MuLV clearance of nanofiltration process, >4 lg copies reduction was shown in virus load between pre- and post-treatment, consistent with the result of cytopathic effect assay. When applied to evaluate composite cation-exchange chromatography, the RT-qPCR method detection results reflected the distribution of X-MuLV particles throughout the process.Conclusion The RT-qPCR method established for X-MuLV has good sensitivity, specificity, accuracy, precision and robustness, applicable for virus clearance processes validation of biological products.

As an important nucleic acid quantification technology in molecular biology, digital PCR has been widely used in the fields of pathogen detection, gene editing detection, and disease diagnosis in recent years due to its advantages of high sensitivity, accuracy, and specificity. This article reviews the basic concepts and principles of digital PCR, as well as the application progress of digital PCR in the field of biological detection, in order to provide a reference for researchers to choose appropriate detection methods.

Objective To investigate the hemolysis and vascular irritation of human coagulation factor Ⅸ (FⅨ) products.Methods According to the provisions of the Technical Guidelines for Studies on Irritation, Sensitization, and Hemolysis of Drugs issued by the Center for Drug Evaluation of the National Medical Products Administration, for FⅨ products, the in vitro test tube method was used to evaluate the hemolytic potential of the drug, and the self-control method with left-right side of the same animal was adopted to conduct the rabbit irritation test by auricular vein injection of 1.6 mL/kg FⅨ or saline for 7 d.Results At a concentration of 50 international unit（IU）/mL, FⅨ did not induce hemolysis or erythrocyte agglutination. During the drug administration period and the recovery period after drug withdrawal in rabbits, no significant abnormalities were observed in general condition, and the vascular irritation scores were all 0. Fourteen days after drug withdrawal, the vascular irritation scores of rabbits remained 0. At 96 h after the last administration and at the end of the 14 d recovery period, the mean pathological scores of rabbits were both 0.Conclusion FⅨ at a concentration of 50 IU/mL does not exhibit hemolytic activity or vascular irritation in rabbits.

Objective To compare the digestive effects of trypsin and trypsin-like enzyme TrypLE on Vero cells, 2BS cells, and MDCK cells, in order to determine their suitability for cell culture and digestion processes.Methods After trypsin solution and TrypLE were used to digest Vero cells, 2BS cells, and MDCK cells, key parameters such as digestion time, dispersion effect, residue after digestion, cell count, and cell viability were analyzed to compare the effects of different digestive enzymes. Digested cells were then inoculated into cell factories and basket bioreactors for cultivation. During the cultivation period, cell morphology and confluence in cell factory were observed daily, and the adhesion effect of cells and glucose consumption in basket bioreactor were measured.Results There were no statistically significant differences in digestion time, cell count, or cell viability of Vero cells and 2BS cells between trypsin and TrypLE digestions (t=﹣0.46-3.46, P=0.074-0.885), and dispersion effect and residue after digestion were similar. However, the digestion time for MDCK cells using TrypLE was statistically significantly longer than using trypsin (t=13.28, P＜0.01), and the digestion effect of TrypLE was inferior to that of trypsin. When these 3 types of cells were re-cultured in the cell factory after digestion, they all had regular shapes and good confluence. When re-cultured in the basket bioreactor, the adhesion rates of 3 types of cells within 3 h were all above 80%, and there was no statistically significant difference in the total glucose metabolism (t=﹣1.05-1.01，P=0.405-0.783).Conclusion TrypLE can replace trypsin for the digestion process of Vero cells and 2BS cells, while trypsin is more effective for digesting MDCK cells.

Objective To evaluate the non-clinical safety comparability of Reassortant Rotavirus Vaccine, Live, Oral, Hexavalent (Vero Cell) produced in pilot and commercial facilities by assessments of single- and repeated-dose toxicity.Methods SD rats were administered by gavage with vaccines produced in pilot and commercial facilities, respectively, at 6 mL dosage, while the proposed clinical dosage was 2.0 mL. The long-term toxicity of the vaccine was assessed through 4 repeated intragastric administrations at doses of 2 or 6 mL per rat on day 1, 15, 29, and 43, with a 4-week recovery period. Results In the single-dose toxicity test, maximum tolerated dose was 6 mL per rat for vaccines produced in both facilities, which was 50-200 times the intended human dose converted by body weight. The body weight gain from day 0—14 and mean daily food intake of rats that received vaccines produced in pilot and commercial facilities were simliar to control groups， which were 33-44 g and 18-20 g, respectively, for female rats and 109-123 g and 25-29 g, respectively, for male rats. In the repeated-dose toxicity test, the no-observed-adverse-effect level was 6 mL per rat. The trends of body weight, mean daily food intake and body temperature in low- and high-dose groups with vaccines produced in pilot and commercial facilities were all simliar to control groups. The blood and biochemical indicators of animals did not show abnormal changes related to different facilities.In equivalent dose vaccine satellite groups, there were no statistically significant differences in seroconversion rate (P=0.474-1.000) and geometric mean titer (GMT) (t=0.05-0.85,P=0.442-0.965) of serum IgG antibodies against rotavirus, and the change trend was consistent. The peak of IgG antibody seroconversion rates and GMTs (1∶lgx) in low- and high-dose vaccine (produced in pilots facilities) satellite groups were 80.0%, 80.0% and 1.61,1.81, while the corresponding vaccine （produced in commercial facilities） satellite groups were 90.0%,100.0% and 1.79,1.91,respectively. The organs and histopathological sections of animals did not show abnormal changes related to different facilities.Conclusion Reassortant Rotavirus Vaccines, Live, Oral, Hexavalent (Vero Cell) produced in pilot and commercial facilities both show good safety in SD rats and are comparable in single-and repeated-dose toxicity studies.

Objective To investigate the physicochemical properties of aluminum phosphate adjuvants subjected to different numbers of autoclaving cycles and to evaluate the effect of aluminum phosphate adjuvant with varying autoclaving cycles on immunogenicity of type 6A pneumococcal polysaccharide-tetanus toxoid (TT) conjugate (6A-TT).Methods Self-prepared aluminum phosphate adjuvants were autoclaved 1, 2, and 3 times, respectively. The particle size and distribution, pH, isoelectric point, free phosphate content, phosphorus-to-aluminum molar ratio, and protein adsorption rate were examined. Additionally, the effect of aluminum phosphate adjuvants treated with different autoclaving cycles, when combined with 6A-TT, on the specific antibody titers in mice was assessed.Results After autoclaving 1, 2, and 3 times, the 50% particle distribution diameters (Dv50s) of aluminum phosphate adjuvant were 5.04, 5.22, and 5.12 μm, while Dv90s were 9.20, 9.87, and 10.60 μm, respectively. The pH values were 4.6, 4.3, and 4.3, while the isoelectric points were 5.48, 5.36, and 5.29, respectively. The free phosphate content were 4.12%, 4.79%, and 4.86%, and the phosphorus-to-aluminum molar ratios were 1.012, 0.972, and 0.944, respectively. The adsorption rates of aluminum adjuvant to TT were 92.7%, 87.8%, and 85.7%, and to 6A-TT were 93.8%, 91.6%, and 90.1%, respectively. The aluminum phosphate adjuvant, after autoclaving 1, 2, and 3 times, was able to adsorb 6A-TT and induce antibody production in mice, with geometric mean titers of 1 213, 985, and 1 056, respectively.Conclusions Increasing the number of autoclaving cycles leads to decrease in the isoelectric point and pH of the aluminum phosphate adjuvant, increase in free phosphate content, and reduction in the phosphorus-to-aluminum molar ratios and protein adsorption rate. It may also have an effect on the immunogenicity of 6A-TT pneumococcal polysaccharide antigen in mice.

Objective To investigate the epidemiological characteristics and breakthrough cases of varicella in Xuancheng City, and provide a basis for epidemic prevention and control.Methods The information of varicella cases in Xuancheng City from 2021 to 2023 was collected through the varicella epidemic report card of China Information System for Disease Control and Prevention, and the epidemiological characteristics were analyzed by descriptive epidemiological method.Results A total of 6 979 varicella cases were reported in Xuancheng City from 2021 to 2023, with an average annual reported incidence rate of 93.44 per 100 000 (χ²=122.80, P＜0.001). The incidence of varicella had significant seasonality, with a small peak from May to July and the highest peak from October to January of the following year. The three counties (cities) with highest average annual incidence rates were Jixi County, Ningguo City, and Jingxian County.The number of male cases was slightly higher than that of female cases, the age of onset was mainly under 14, and occupations were mainly students and kindergarteners.There were 2 652 breakthrough cases, accounting for 38.00% of the total cases. There were more breakthrough cases in males than in females, and the proportion of breakthrough cases in autumn was the largest. In 11 years after vaccination, longer time since vaccination was associated with more breakthrough cases.Breakthrough cases in clustered epidemic accounted for 68.45% of related cases, significantly higher than that in sporadic cases. Breakthrough cases were mainly students aged 7-13.Conclusions The incidence of varicella in Xuancheng City is on the rise, and schools and kindergartens are the key places for the prevention and control of varicella. It is nessesary to prevent the occurrence of cluster epidemic, increase 2-dose varicella vaccination, and reduce the occurrence of breakthrough cases. CDC should strengthen epidemic monitoring, review epidemic report, carry out targeted training and health education, and establish effective immune barriers to reduce the incidence of varicella.

Objective To screen differences in abnormal toxicity test (ATT) for same biological product using animals from different suppliers and to explore the scientific and rational nature of ATT by additional blood routine analysis.Methods With reference to Chinese pharmacopoeia 2025 edition (volume Ⅲ) general rule 1141 “Abnormal toxicity test method”, the differences between animals from different suppliers were compared by analyzing the weights and blood routine results of animals after immunization with different vaccines.Results The body weight growth trend of NIH mice from supplier B was statistically significantly lower than that of the current supplier (supplier A) in both control and vaccine groups (t=2.64-7.07, all P<0.05), and the difference between suppliers C and A was not significant. Using the blood routine results of NIH mice from supplier A as a reference, blood routine results of mice from supplier B had significantly more red blood cells in both vaccine groups (t=2.44-4.07, all P<0.05) and showed significant differences in many white blood cell indicators (t=2.31-5.96, all P<0.05), while that of supplier C only showed significant differences in a few indicators. The body weight growth trends of Hartley guinea pigs from 3 suppliers (B, D, E) remained the same in both control and vaccine groups, and there was no significant difference in blood routine results.Conclusions NIH mice from supplier B have significantly different quality from those from suppliers A and C, while the quality of Hartley guinea pigs from suppliers B, D, and E is similar. The difference in animal quality directly affects the determination of ATT results, and the rationality of using ATT as a testing method for vaccine batch release is worth exploring.

Objective To evaluate the application effects of 3 types of sheet carriers for the production of Sabin strain inactivated poliovirus vaccine (sIPV) in order to provide basis for carrier selection in sIPV production process.Methods A 15 L basket bioreactor system was adopted to carry out Vero cell culture and prepare Sabin strain poliovirus liquid with 3 sheet carriers A, B, and C, respectively. The cell attachment rate of Vero cells inoculated on the sheet carriers within 0-3 h, the glucose metabolism characteristics during the 8-day culture cycle, the virus titer and D-antigen content of the virus harvest were investigated and compared.Results Cell attachment rates of carriers A, B, and C were all >90% at 3 h after inoculation. Cells in all 3 groups presented a typical S-shaped glucose consumption curve. The cumulative glucose consumptions of A and B were (191.1±7.6) g and (192.9±5.3) g, with no statistically significant difference (F=43.23, P=0.060), while both were statistically significantly higher than that of C at (153.1±4.4) g (F=43.23, P<0.001). D-antigen contents of A and B were (773.3±10.8) and (777.0±12.5) D-antigen unit (DU)/mL, and virus titers were (8.1±0.2), (8.2±0.1) lgCCID₅₀/mL,respectively, with no statistically significant difference (F=209.00, 27.30; P=0.946, 0.708). In contrast, the two indicators of group C at (598.3±14.2) DU/mL and (7.2±0.2) lgCCID₅₀/mL,were lower than those of group B with statistically significant differences (F=209.00,27.30; P<0.001).Conclusion Carriers A and B both exhibit clear advantages in maintaining the metabolic activity of Vero cells and promoting poliovirus replication.

Immunometabolism is an emerging field that explores the interactions between immune cells and metabolism. In recent years, the role of immunometabolism in autoimmunity and autoimmune diseases has become a growing area of research, focusing on how cellular metabolism influences immune cell function and responses. It is currently understood that immune cell signaling and differentiation processes can determine immune cell behavior, control numerous metabolic pathways and thereby affect immune cell function and responses. This article primarily discusses the metabolic alterations in immune cells in systemic lupus erythematosus, type 1 diabetes and rheumatoid arthritis, elucidating how immunometabolism influences disease onset, exacerbation, or remission, and summarizes drugs that modulate immunometabolism to potentially control immune responses in autoimmune diseases and their research progress.

Seasonal influenza poses a persistent threat to global public health, imposing a significant disease burden particularly on high-risk population such as the elderly, children, pregnant women, chronic diseases patients, and immunocompromised people. Although the effectiveness of existing influenza vaccines is contingent on their match with circulating strains, their protective benefits for high-risk population are well-supported by epidemiological evidence. Currently, influenza vaccination coverage in China remains considerably lower than developed countries. There is an urgent need to enhance vaccination rates among high-risk population through policy support and technological innovation to reduce the disease burden of influenza. This article systematically reviews the protective evidence of influenza vaccines in high-risk population, focusing on the effectiveness and applicability of different vaccine types (inactivated, live attenuated, recombinant protein, and adjuvanted vaccines) in the high-risk population. It also examines the effects of repeated vaccination, the role of herd immunity, and the optimization of vaccination strategies, providing references to improve protection in high-risk population.

Objective To analyze the incidence and distribution characteristics of adverse events following immunization (AEFIs) associated with diphtheria-tetanus-acellular pertussis vaccine (DTaP) before and after the implementation of new immunization strategy under the National Immunization Program in Anhui Province, and to provide evidence for evaluating the safety of the new strategy.Methods AEFI surveillance data in Anhui Province during 2024（before strategy adjustment） to 2025（after the new strategy implemention on January 1, 2025） were collected through the Immunization Program Information Management System of the Chinese Center for Disease Control and Prevention, and concurrent vaccination data were obtained from Anhui Immunization Program Information Management System. Descriptive epidemiological methods were used to analyze the distribution and incidence of AEFI. The Chi-square test or Fisher’s precision probability test were used to compare the data before and after the policy adjustment.Results From 2024 to 2025, a total of 2 375 028 doses of DTaP were administered in Anhui Province, with 4 958 AEFI cases reported. The overall incidence was 208.76/100 000 doses. Following the strategy adjustment, the incidence of general reactions increased from 186.69/100 000 doses to 221.37/100 000 doses (χ²=32.50, P<0.001)，while the incidence of adverse reactions (rare/serious) decreased from 0.66/100 000 doses to 0.20/100 000 doses, representing a reduction of approximately 69.70%. After advancing the first dose to 2 months of age, the AEFI incidence decreased from 170.57/100 000 doses to 119.65/100 000 doses (χ²=24.76, P<0.001). For the 6-year-old dose switched from the diphtheria and tetanus vaccine to DTaP, no adverse reactions were observed, with incidence of 0.00/100 000 doses (95% confidence interval: 0.00-0.83/100 000 doses ).Conclusions DTaP AEFIs in Anhui Province are predominantly general reactions, with extremely low adverse reaction rates. Advancing the first dose to 2 months of age significantly reduces the AEFI incidence, facilitating the early establishment of immune barrier. With no new safety risks monitored during this surveillance period after switching to DTaP for the 6-year-old booster, core safety indicators perform well during the study period. The new 5-dose DTaP strategy (at 2, 4, 6, 18 months and 6 years of age) demonstrates a good safety profile within the observation period.

Objective To analyze the characteristics of adverse events following immunization (AEFIs) with varicella live attenuated vaccine (VarV) in Anhui Province from 2016 to 2025.Methods Data on VarV AEFI cases in Anhui Province from 2016 to 2025 were collected through the China Disease Control and Prevention Information System, and the number of VarV vaccination doses were collected through the Anhui Immunization Program Information System. Descriptive epidemiological methods were used to analyze the incidence and distribution characteristics of AEFIs.Results A total of 2 408 VarV AEFIs were reported in Anhui Province from 2016 to 2025, with an average annual incidence of 329.46 per million doses. The incidence of common reactions and rare adverse reactions were 308.46 per million doses and 17.80 per million doses, respectively. Among all cases, 53.74% were male, 64.70% were ≤1 year old, 61.17% were from northern Anhui Province, 78.15% occurred within 24 h, and 77.24% were from the first dose. Among rare adverse reactions, anaphylactic rash accounted for 90.77%, with an average annual reported incidence of 16.16 per million doses.Conclusion VarV AEFIs are mainly common reactions in Anhui Province, with extremely rare adverse reactions, which shows an overall good safety profile of VarV.

Objective To analyze adverse events associated with rabies vaccine for human use (RVFHU) based on data from the US Vaccine Adverse Event Reporting System (VAERS) and to evaluate the safety profile of RVFHU.Methods RVFHU-related reports were retrieved from the VAERS database for the period from January 1, 2019 to September 15, 2023. Descriptive statistical analysis was employed to characterize the distribution of adverse events, and disproportionality analysis was applied to detect vaccine safety signals.Results A total of 794 adverse event reports (comprising 2 606 clinical symptoms) were included. Product use issues were the most frequently reported clinical symptoms. In terms of System Organ Class, symptoms related to general disorders and administration site conditions accounted for the highest proportion. Regarding the composition ratios of general adverse events and serious adverse events by vaccination dose, the difference was statistically significant (χ²=10.25, P<0.05), whereas no statistically significant differences were observed for gender, age, or time of occurrence (χ²=0.32, 2.58, 2.02, respectively; P>0.05). Disproportionality analysis identified 59 positive safety signals, involving product use issues, immune system disorders, psychiatric disorders, and other aspects.Conclusions Adverse events associated with RVFHU are predominantly product use issues or transient and mild reactions. The positive safety signals detected are limited in number and require further investigation and validation.

Objective To establish and validate a cation exchange chromatography detection method for adalimumab charge variants.Methods An exchange column (4 mm×250 mm) with weak cations of carboxylic functional groups was used as the stationary phase. 0.01 mol/L disodium hydrogen phosphate solution, pH 7.5 was used as mobile phase A, and 0.01 mol/L disodium hydrogen phosphate + 0.5 mol/L sodium chloride solution, pH 5.5 was used as mobile phase B for gradient elution. The content of lysine variants and acidic components were calculated by area normalization method. Specificity, repeatability, intermediate precision, linearity, accuracy, and robustness of the method were validated.Results Adalimumab target peak and solvent peak were observed. For 6 repeated tests, relative standard deviations (RSDs) of all components were all ＜2%. With different experimenters and test equipment, RSDs of each component were all ＜ 2%. The quantification limit (signal-to-noise ratio＞10) and detection limit (signal-to-noise ratio＞3) of peaks between lysine variants were 100 μg/mL and 30 μg/mL, respectively. Within the quality standard limits of Wuhan Institute of Biological Products Co., Ltd., the linearities of all components at different concentrations were good, with coefficients of determination all＞0.99. Recovery rates were within 80%-115% for the first acidic region peak, the intermediate peak between lysine variants, and lysine variant 2 peak, and 85%-110% for the second acidic region peak, main peak, and lysine variant 1 peak. Analytical results met the specified requirements and exhibited acceptable consistency under different mobile phase pH and NaCl concentration, flow rate, column temperature, and chromatographic column lot numbers.Conclusion The method established has good specificity, repeatability, intermediate precision, linearity, accuracy, and robustness, and is suitable for the determination of the purity of adalimumab charge variants.

Objective To identify the key binding sites and preliminarily evaluate the biological functions of fully humanized monoclonal antibody (mAb) 23F4 targeting sialic acid-binding immunoglobulin-like lectin 15 (Siglec-15).Methods Alanine scanning mutagenesis was performed on Siglec-15 to identify key binding sites of 23F4. In vitro biological activity was assessed via ELISA and flow cytometry. In vivo antitumor efficacy was evaluated using 2 murine tumor models.Results The critical binding sites of 23F4 were identified as residue R71/H72. In vitro, 23F4 exhibited superior binding activity with median effective concentration of 68.42 ng/mL, and outperformed control antibodies in binding Siglec-15-positive cells particularly at medium-to-low concentrations. 23F4 blocked Siglec-15-receptor interaction, reversed T-cell proliferation suppression, and suppressed tumor growth in both murine tumor models.Conclusion As a Siglec-15-targeting mAb with novel binding sites, 23F4 effectively alleviates Siglec-15-mediated immunosuppression.

Objective To evaluate the transportation stability of adalimumab injection.Methods The worst-case transport conditions were simulated by conducting a round-trip road transport from Wuhan to Harbin, to assess drug stability under non-temperature factors. Destructive tests, including extreme temperature exposure and temperature cycling, were employed to simulate post-cold-chain-break scenarios. Stability under temperature excursions was evaluated by testing routine quality attributes of adalimumab injection.Results The primary packaging of all 3 batches remained intact post-transportation, with 0% breakage rate. All tested attributes complied with the company’s registered specifications, showing no statistically significant differences compared to pre-transport samples (t=﹣3.23-4.00,P=0.057-0.840). After storage at (40±2) °C or (﹣20±5) °C for 7 d, all attributes for 3 batches met the company’s registered specifications. However, monomer content (determined by size exclusion chromatography) and purity (determined by ion chromatography) exhibited statistically significant differences (t=﹣142.00-97.00，P=0.000-0.020) between samples stored at (40 ± 2) °C and those stored at 2-8 °C or (﹣20±5) °C. No statistically significant differences (t=0.70-3.46，P=0.074-0.556) were observed in any attributes between samples stored at (﹣20±5) °C for 7 days and those stored at 2-8 °C. All attributes met the company’s registered specifications after 3 freeze-thaw cycles, with no statistically significant differences compared to samples not subjected to freeze-thaw testing (t=﹣4.06-1.73，P=0.056-0.478).Conclusion Under the experimental conditions, non-temperature factors during long-distance transportation have no impact on drug quality, and short-term temperature excursions have minimal effect on the quality of adalimumab injection.

Objective To validate the method of cell species identification (multiplex PCR) and apply it to the identification of cells from various species.Methods Cell genomic DNA was extracted by cell species identification detection kit (multiplex PCR), and the target gene was amplified by mixed primers. Multiplex PCR combined with agarose gel electrophoresis was used to identify species and detect cross-contamination according to the band size and number of amplified products, and the detection limit, specificity and durability to 5 different sources of cells were investigated.Results The detection limits of multiplex PCR method were 500 cells for Hep-2 cells (human), canine kidney cells, Vero cells (African green monkey) and L929 cells (murine), and 5 000 cells for Chinese hamster ovary cells. When each cell type was designed with cells from different sources for cross-contamination, both main cells and contaminant cells as low as 1‰ level were detected. When the genomic DNA extracts of 5 kinds of cells were stored at ﹣18 ℃ for 1, 3 and 7 d, the amplification results were consistent.Conclusion Multiplex PCR method has high sensitivity, strong specificity, rapidity and convenience, which is helpful to improve the quality control level of cell substrates for the production of biological products.

With the rapid development of microneedle technology, microneedle has gradually become a new vaccine delivery method, which has advantages such as minimally invasive, painless, self-administration, dose-saving, and enhanced immune response. The evaluation of immunogenicity is crucial for vaccine research and development. However, there is still no systematic summary of immunogenicity evaluation method of microneedle vaccines. This article summarizes the characteristics of immune responses to microneedle vaccines, systematically reviews and discusses various methods and key indicators for evaluating the immunogenicity of microneedle vaccines, as well as explores and prospects emerging immunogenicity evaluation methods in microneedle vaccines.

Objective To develop and validate the isoelectric point (pI) detection method for recombinant human interferon α1b (rhIFNα1b) by imaging capillary isoelectric focusing electrophoresis (iCIEF).Methods Based on The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use Q2 (R2) requirement, iCIEF was used to detect the pI of rhIFNα1b, and the concentration of urea added to sample and isoelectric focus time were determined.The developed method was verified for specificity, accuracy, repeatability, intermediate precision and durability.Results In iCIEF, the focusing was performed at 3 000 V for 3 min, and 1 mol/L urea was added to treat samples. Blank control (acetate buffer) showed no interference to the determination of pI of rhIFNα1b. Isoelectric focusing of rhIFNα1b generated 2 peaks. The pIs of peak 1 and 2 were approximately 5.4 and 5.5, respectively. The method had good specificity, accuracy, repeatability, intermediate precision, and durability.Conclusion This established iCIEF method can be used for the detection of the pI of rhIFNα1b, providing a reference for the quality control of this product.

With the increasing frequency of emerging infectious diseases globally in recent years, the response practices to previous public health crises indicate that a rapid vaccine response is crucial to effective threats containing from pathogens with large-scale epidemic potential. In 2021, the Coalition for Epidemic Preparedness Innovations (CEPI) proposed the strategic plan CEPI 2.0, also known as the 100 Days Mission. The six pillars of the plan, namely establishing a prototype vaccine library, building a global clinical trial network, identifying early immune biomarkers, developing rapid manufacturing and validation capabilities, deploying early outbreak surveillance systems, and strengthening biosafety and biosecurity capacity, aim to achieve preliminary authorization and mass production readiness for target vaccines within 100 days after confirmation of a pandemic pathogen.This paper provides a systematic review and interpretation of the scientific framework of the plan, seeking to offer reference and insights for China to better respond to potential epidemic risks and pandemic occurrences.