Hantavirus (HV) infection can cause two acute infectious diseases, hemorrhagic fever with renal syndrome and hantavirus cardiopulmonary syndrome, which have spread and led to high mortality rates around the world, becoming a global public health problem. Currently, there are no specific drugs or methods for treatment of HV. Vaccines have become a effective and economical means to prevent potential pandemics. At present, HV vaccines mainly include inactivated vaccines, subunit vaccines, virus vector vaccines, virus-like particle vaccines, DNA vaccines, etc. They mainly use viral structural protein glycoprotein and nucleocapsid protein as immunogens to induce humoral and cellular immune responses to eliminate HV infected cells. This article briefly reviews the types, advantages and disadvantages, mechanisms of action, as well as research progress of HV vaccines at home and abroad.

According to difference of principle, virus detection methods can be divided into electron microscope method, cell culture method, immunoassay, nucleic acid testing, etc. Electron microscope method with a high resolution can show the structure of the viruses directly, which is suitable for the study of unknown virus. Cell culture methods can obtain the virus activity and detect virus titers. Immunoassay has strong specificity, low cost, and high speed, which is suitable for large-scale population screening. Nucleic acid detection methods allow simultaneous detection of multiple viruses with high sensitivity and strong specificity. Researchers should make a trade-off in the choice of method. This article discusses advantages and disadvantages of virus detection methods and applications, to provide reference for development of more accurate and efficient virus detection, prevention and control strategies.

Protein nanoparticles, as antigen display carriers, can effectively help to improve the stability and immunogenicity of antigens due to their similar size to natural pathogens. Clinical trials have shown that protein nanoparticles can be used in the development of novel vaccines. This paper reviews the immunological properties of protein nanoparticle vaccine and the application in vaccine research and development, aiming to provide reference for further research of protein nanoparticle.

Recombinant virus vector vaccine is a kind of vaccine which uses virus as vector to deliver target antigen. The technical route of virus vector was mostly used for animal vaccines and the pathogen vaccines which were difficult to develop with traditional technical routes in the early stage. Among the five technical routes for the research and development of vaccines against COVID-19 in China, the recombinant virus vector vaccine was approved for marketing and played an important role in the prevention and control of the epidemic because of its advantages of simultaneously producing strong humoral and cellular immunity, being vaccinated through mucosal pathway, and completing the immunization program with one dose. However, due to the pre-existing immune problems of this vaccine, such as factors that may reduce the protective efficacy of the vaccine, and the increase of safety risks in order to enhance immunogenicity, there are few recombinant virus vector vaccine products on the market. With the development of genetic engineering and molecular biology technology, the variety of virus vectors is increasing day by day, and it has been applied in many vaccines. In this paper, the research and development status of recombinant virus vector vaccine is reviewed.

IL-32 is a cytokine with 9 subtypes widely expressed in human body. IL-32 can regulate the function of immune cells and other cells through various pathways, cooperate with multiple cytokines to participate in the progress of inflammation, and directly or indirectly play a pro-inflammatory or anti-inflammatory role. IL-32 also has important impact on tumor initiation, invasion and migration, and its functional differences may be related to different subtypes and complex tumor microenvironment. This article aims to elucidate the role and related mechanisms of IL-32 in inflammation and tumors.

Rabies caused by rabies virus is a lethal infectious zoonosis that infects the nervous system. The clinical conditions mainly include hydrophobia, salivation, mania or excitement. At present, there is no specific therapy for rabies and the disease is 100% lethal. The rabies vaccine remains the most effective strategy for rabies prevention and control. This article reviews the epidemiology of rabies, rabies vaccine strains and the research progress of human rabies vaccine in China, in order to provide a theoretical basis for optimizing rabies prevention and control strategies.

Lipid nanoparticles (LNPs), with spherical vesicle structure composed of one or multiple phospholipid bilayers, are currently the most effective non-viral delivery vehicle in clinical research and application, mainly for the delivery of chemical drugs and nucleic acid molecules. Since the outbreak of COVID-19, mRNA vaccines standed out due to their simplicity of design and manufacture, low immunogenicity, and rapid large-scale production. As an important component, LNPs play an important role in the effectiveness and stability of mRNA vaccines against COVID-19. In this review, the structure, characteristics, application and quality control of the widely used LNPs are summarized, in order to provide a deeper and broader understanding to mRNA vaccine delivery systems and further promote the rapid development of mRNA vaccines and drugs.

Objective　To evaluate the passage stability of pneumococcal subcultures of all serotypes for pneumococcal polysaccharide conjugate vaccine in a liquid medium.Methods　The culture duration of each generation of pneumococcus in a liquid environment was compared. The characterization of bacteria and capsular polysaccharide antigen gene sequences of the 15th generation (P15) and P4 (control group) strain were analyzed. Then, P15 was fermented and purified according to the production process, and the yield and quality of capsular polysaccharide antigen were analyzed by determination methods and compared with P4.Results　In P7-P15, the culture duration of each generation was similar, and there was no difference in strain identification and antigen gene sequence between P15 and P4, with the sequence similarity of 100%. After the fermentation of P15 strains, there was no apparent difference in the growth of cells, the yield of capsular polysaccharides and the related indexes of capsular polysaccharides compared with P4, and the verification indexes were in line with Cinese pharmacopoeia standards.Conclusion　Pneumococcal subcultures of all serotypes for pneumococcal polysaccharide conjugate vaccine have good passage stability in a liquid environment.

Cancer is one of the leading causes of death worldwide, and its etiology and pathogenesis are still not well understood. Human cytomegalovirus (HCMV) is a type of herpesvirus that encodes multiple oncogenes, activates various important signaling pathways related to cancer, and promotes cancer initiation and development after infection. Studies on breast cancer, glioblastoma, and muscle sarcoma etc have provided important evidence in this regard. This article reviews the molecular mechanisms and clinical researches of HCMV-induced carcinogenesis, aiming to provide new insights and targets for clinical prevention and treatment of HCMV infection.

Objective　To investigate the IgG antibody levels against Neisseria meningitidis serogroups among children after group A and C meningococcal polysaccharide conjugate vaccine (MPCV-AC) was included in the immunization program in Anhui province for 11 years, to provide reference for further optimizing of the immunization strategy against epidemic meningococcal meningitis.Methods　A multi-stage random sampling method was employed to select healthy children under 11 years old from six cities in Anhui province. Questionnaires were conducted and blood specimens were collected.ELISA was used to detect the levels of antibody against groups A, C, Y and W135, and the data were statistically analyzed.Results　A total of 1 002 children were investigated. The proportions of children with protective antibody concentrations against serogroups A, C, W135 and Y were 89.82%, 92.12%, 31.04% and 66.87%, respectively. The median antibody concentrations were 9.93, 10.55, 4.63 and 7.75 μg/ml, respectively. The positive rates of serum anti-Nm IgG antibodies in four different age groups were statistically significantly different, with χ² values of 27.75, 17.70, 132.00 and 122.55, respectively, and P＜0.05. Children who received booster immunization with group A, C, Y, W135 meningococcal polysaccharide vaccine had significantly higher positive rates of anti-Nm IgG antibodies in W135 and Y groups compared to those who received MPCV-AC（χ² values were 10.30 and 38.25, P＜0.05）.Conclusions　The IgG antibody levels against serogroups A and C of Nm among children aged 0 to 11 years in Anhui province are relatively high after the inclusion of MPCV-AC in the immunization schedule, showing significant effectiveness in meningococcal prevention and control. However, the IgG antibody levels against serogroups W135 and Y remain relatively low in this population, indicating a need for strengthened educational efforts to improve the vaccination rate of meningococcal polyvalent vaccine.

Objective　To compare the effects of different signal peptide sequences on secretory expression of influenza virus hemagglutinin (HA) in Sf9 insect cells.Methods　The HA of H1N1 subtype influenza virus A/Victoria/2570/2019 (IVR-215) was used as a target.The full-length HA gene sequence was selected and naïve signal peptide, honeybee melittin signal peptide, glycoprotein 67 signal peptide（Gp67sp）, chitinase signal peptide, and human azurocidin gene signal peptide were connected to the 5´terminal of the gene， respectively. The genes were cloned into the genome of the pFastBac1 transfer plasmid， and five recombinant baculoviruses were constructed by the Bac-to-Bac system. The viral titers were detected by BacPAK^TM qPCR titration kit. The recombinant baculoviruses were used to infect Sf9 cells at different multiplicities of infection, and the cell supernatants were harvested at different time points after infection, and the protein expression levels were detected by immunoblotting to optimize the conditions for HA secretory expression.Results　The titers of the five recombinant baculoviruses harvested were all ＞10⁶ PFU/ml, and the Sf9 cells were able to correctly express and secrete HA, with a relative molecular mass of about 80 000. The amount of HA expressed was the highest when mediated by Gp67sp.The highest expression of HA was achieved when the recombinant baculovirus AcMNPV-Gp67sp-HA infected Sf9 cells for 96 h with multiplicity of infection 5.0 or 10.0, and the hemagglutination efficiency was 256 hemaqqlutinetion units/50 μl.Conclusion　All signal peptides tested effectively guide the secretory expression of influenza virus HA protein in Sf9 cells, among which Gp67sp is particularly advantageous for the efficient secretory expression of HA.

Objective　To structurally optimize and preliminarily identify the biological activity of anti-tumor necrosis factor receptor superfamily member 4 (OX40) monoclonal antibody Ab18.Methods　The species cross-binding activity of Ab18 to OX40 of different species was measured by ELISA. The binding activity of Ab18 to HEK293-hOX40 cells was measured by flow cytometry (FCM). The blocking activity and agonistic activity of Ab18 were measured by reporter gene methods. The Fc fragment of Ab18 was engineered by point mutagenesis technology to change the affinity of the Fc fragment with Fc receptors. ELISA and FCM were used to evaluate the binding activity to human Fcγ receptor ⅡB and human neonatal Fc receptor, and the reporter gene method was used to measure the agonistic activity and antibody-dependent cell-mediated cytotoxicity (ADCC) activity of Ab18 and its mutants.Results　The Ab18 antibody had species cross-binding activity with mouse, rat and rhesus monkey OX40s, and stronger binding activity and stronger blocking activity with HEK293-hOX40 than the control antibodies GBR830 and KHK4083. The median effect concentration (EC₅₀) value of binding activity of Ab18 with HEK293-hOX40 was 366.9 ng/ml, and the half maximal inhibitory concentration value of blocking activity was 657.3 ng/ml. In addition, Ab18, GBR830 and KHK4083 showed agonistic activity. The ADCC activity EC₅₀ value of Ab18 was 25.7 ng/ml, and the agonistic activity EC₅₀ value was 14.9 ng/ml. The mutated Ab18 had lower agonistic activity, better binding activity with human neonatal Fc receptor, consistent ADCC activity, and good biological activity.Conclusion　The Ab18 antibody is successfully optimized, showing better biological activity.

Hypoalbuminemia is an independent predictor of poor prognosis in major abdominal gastrointestinal surgery. To avoid the occurrence of hypoalbuminemia after abdominal surgery, the application of human serum albumin is relatively common in patients with hypoalbuminemia after abdominal surgery due to insufficient level of albumin. However, there is still controversy regarding the clinical benefits of using human serum albumin after abdominal surgery.This article reviews the current role, pathological and physiological changes, clinical treatment benefits, and nutritional value of albumin in the perioperative period of abdominal surgery, in order to strengthen clinical physicians' correct understanding and rational application of human albumin, in turn to facilitate postoperative management of patients and reduce medication costs.

Objective　To establish and validate an imaging capillary isoelectric focusing electrophoresis(icIEF) method for the detection of charge variants in the therapeutic monoclonal antibody SIBP-A. Methods　The icIEF conditions were first optimized by adjusting urea concentration, ampholyte range, ampholyte concentration and focusing time. The optimized method was then applied to analyze the charge variants of SIBP-A, and its specificity, stability indicating property,repeatability, intermediate precision, linearity and range, accuracy, limit of quantification and stability of processed samples were verified. Results　The optimized icIEF conditions included 3 mol/L urea, 1% MC gel, and a 1:1 mixture of two ampholytes (Pharmalyte 3-10 and Pharmalyte 8-10.5) as carriers, 1 kV prefocusing voltage with 1 min prefocusing time, and 3 kV focusing voltage with 6 min focusing time. In the specificity experiment, the blank group registered no significant interference peak at the sample peak while the control group exhibited normal peaks and charge variants. In the stability indicating experiment at 40 ℃ for 7 d, the sample showed normal peaks but with significant changes in charge isoforms, which were distinct from the control sample. The repeatability experiment demonstrated relative standard deviation (RSD) values of 0.48%, 0.00%, and 4.03% for the acidic, main, and basic peak relative percentage contents, respectively, and an RSD of 0.012% for the main peak's isoelectric point. Precision experiments showed RSD values of 0.89%, 0.45%, and 6.14% for the acidic, main, and basic peak relative percentage contents, respectively, with an RSD of 0.072% for the main peak's isoelectric point. In the protein range of 0.50-1.50 mg/ml, the coefficient of determination of the linear regression cure of the total peak area was 0.996 8 with good linearity, and the recovery rate of each protein concentration were 95.03%-104.35%. The limit of quantitation was 0.003 0 mg/ml.The sample of stability experiments showed RSD values of 0.56%, 0.00%, and 8.61% for the acidic, main, and basic peak relative percentage contents, respectively, with an RSD of 0.056% for the main peak's isoelectric point at 10 ℃. Under room temperature condition, the charge variants changed from 3 h. Conclusion　The developed icIEF method effectively detects and characterizes charge variants and isoelectric points in the therapeutic monoclonal antibody SIBP-A. The specificity, stability indicating property, repeatability, intermediate precision, linearity and range, accuracy, limit of quantitation and stability at 10 ℃ are all satisfactory, making it suitable for release testing and stability testing of the antibody.

Quantitative PCR (qPCR) is widely used in various fields such as mRNA expression, pathogen detection, and viral titration, and is characterized by rapidity, high sensitivity, low contamination rate and reproducibility. This paper mainly reviews the research progress of qPCR in titer detection of various viruses, introduces the appplication of qPCR in detection of some RNA viruses and DNA virus, and compares it with other detection methods, to show the application prospect of qPCR for virus titer detection.

Objective　To study the variation of measles virus titer with temperature and time under specific storage conditions of measles virus bulk and measles-containing vaccines made by cell factory process.Methods　Three batches each of measles virus, mumps virus and rubella virus bulks were prepared according to cell factory process. The measles virus bulks were stored at 2-8 ℃ for 21 d and below -60 ℃ for 15 months for stability analysis. The same batchs of measles virus bulk were used to prepare six batches each of measles attenuated live vaccine (MV), measles and mumps combined attenuated live vaccine (MM) and measles, mumps and rubella combined attenuated live vaccine (MMR). Accelerated stability test at （25±2） ℃ for 6 months and long-term stability test at 2-8 ℃ for 30 months were carried out on 18 batches of vaccines.Measles virus titers of bulk and final products were detected, and the stability of measles virus in different samples at different storage temperatures was analyzed. Minitab 17 analysis software was used to predict the stability period of bulk and final products.Results　In 3 batches of measles virus bulks stored at 2-8 ℃ for 21 d and below -60 ℃ for 15 months, the lowest titers of measles virus were 5.5 and 5.8 lgCCID₅₀/ml, respectively, and met the quality standard of ≥ 5.3 lgCCID₅₀/ml. The lowest measles virus titers of MV, MM and MMR were 4.0, 4.7 and 3.4 lgCCID₅₀/ml, respectively, when 18 batches of vaccines were stored at （25 ± 2） ℃ for 6 months, and were 4.4, 4.5 and 3.8 lgCCID₅₀/ml, respectively, when stored at 2-8 ℃ for 30 months. The measles virus titer in the stability test of final products all met the product quality standards. The stability period of bulk and final products predicted by analysis software was far beyond the corresponding approved effective period.Conclusion　In measles virus bulk and three measles-containing vaccines made by cell factory process, measles virus has good stability.

Objective　To establish a method for the detection of biological activity of human growth hormone (hGH) injection based on hGH promoting proliferation of the rat lymphoma cell line (Nb2-11 cell) and to validate the method.Methods　The proliferation of hGH stimulated cells was measured by luminescent cell viability assay, and the relative potency of hGH was calculated by the 4-parameter logistic regression model. The detection method was established by screening the analysis medium formulae (1%, 5% and 10% horse serum), cell treatment methods (cells starved for 24 h or without starvation treatment) and cell densities (5 000 or 8 000 cells/well). The Nb2-11 cell proliferation method adopted was verified for specificity, accuracy, precision, linearity, durability and range according to the general requirements 9401 of Chinese pharmacopoeia 2020 edition volume Ⅳ.Results　The optimal curve fit was obtained using assay medium containing 5% horse serum, cells starved for 24 h, and a density of 8 000 cells/well. The hGH preparation buffer had no effect on the detection results. The linear regression equation was: y=0.942 8x + 0.072 6, with a coefficient of determination of 0.980 9, and the linear range was 60%-140%. The single experiment recovery rates of different active samples were 70%-130%. The coefficient of variation of the validation experiment was less than 20%.Conclusion　A method for determination of biological activity of hGH injection is successfully developed, which shows good specificity, accuracy, precision, linearity and durability, and can be used for the biological activity detection of hGH injection.

HPV is a type of DNA virus that exists widely in nature, and the persistent infection of HPV can lead to the occurrence of a variety of cancers, especially cervical cancer. The development and application of HPV vaccine is of great significance for the prevention of cervical cancer and other related cancers. In recent years, with the advancement of science and technology, the research of HPV vaccine has made remarkable progress. There are currently three HPV vaccines on the global market including bivalent, quadrivalent and nine-valent vaccines. The bivalent and quadrivalent vaccines can prevent nearly 70% of HPV-related cervical cancers and precancerous lesions, in contrast, the nine-valent vaccine can provide nearly 90% protection. This article reviews the research progress of HPV vaccines.

Objective　To construct a single-chain fragment variable (ScFv) immune library targeting carcinoembryonic antigen-related cell adhesion molecule 5 (CEA), and to obtain antibodies targeting CEA by screening..Methods　The recombinant phage plasmids were constructed by cloning the antibody sequences from mouse spleen into phage plasmids. The plasmids were then electrically transformed into E. coli strain TG1, and rescued by M13KO7 helper phage to obtain a variety of phage antibody libraries from TG1 strain. Using in vitro phage library solid-phase affinity screening method and CEA protein as target, high-affinity antibodies were screened.Results　A murine-derived anti-CEA phage ScFv immune library was successfully constructed, and the library capacity was calculated to be 1.02×10⁸ CFU.ScFv targeting CEA was obtained by screening.Conclusion　The anti-CEA antibody is screened by the phage display technique, laying foundation for the screening of anti-CEA antibody.

Objective　To establish and verify a pharmacokinetic detection method for rabbit anti-human thymus/lymphocyte immunoglobulin (r-ATG) , for detection of the total concentration of r-ATG in the sera of patients after clinical medication.Methods　Mouse anti-rabbit IgG was used to coat the ELISA plate, and horseradish peroxidase-labeled goat anti-rabbit IgG was used as the secondary antibody to establish a double-antibody sandwich ELISA method for the detection of r-ATG content in human serum. The linear range, parallelism, repeatability, intermediate precision, accuracy, specificity, and stability of recovery rates for standard curve concentrations of reference materials at different time points of the method were verified. Clinical blood samples of 5 patients were detected and pharmacokinetic analysis was carried out by the established method.Results　When the initial concentration was 0.25 μg/ml and the dilution times were between 2¹-2⁹, the reference concentration and the absorbance value showed an S curve, the coefficient of determination (R²) was > 0.99, and the recovery rate was 90%-110%. The coefficient of variation (CV) of slope of linear equation between reference sample and three blood samples was < 10%, with R² > 0.99, CV of R² < 5%, showing good parallelism . The repeatability CV and intermediate precision CV were <15%, and the recovery rates were all 90%-110%. The average recovery rates of accuracy verification results were 80%-120%. There was statistically significant difference between the sera of the injected patients, healthy people, and non-injected patients, the t-values were 32.607 and 21.949, respectively, with P <0.01. The average recovery of each concentration in the standard curve was 90%-110% when the reference was stored at 2-8 ℃ for 16 months, and the relative standard deviation of the recovery at different time points of each concentration was less than 15%. The serum concentration of r-ATG reached the highest value on the 5th day after administration, with an average half-life time of (6.5±5.3) d.Conclusion　The established method has good dilution linearity and parallelism, repeatability, intermediate precision, accuracy, specificity between samples, and the recovery rate of each concentration in the reference standard curve at different time points is stable, thus can be used for the detection of serum concentration after clinical administration of r-ATG.

Objective　To elucidate the correlation between three determination methods for lymphocytotoxicity potency of anti-human T lymphocyte immunoglobulin by comparing the titers of anti-human T lymphocyte immunoglobulin immune serum and bulk measured by 3 methods. Methods　The titers of three batches of immune serum and bulk were determined by lymphocytotoxicity test in Chinese pharmacopoeia 2020 edition, peripheral blood mononuclear cell （PBMC） based and human leukemia T cell line （Jurkat cell） based lymphocyte cytotoxicity flow cytometry methods. The 128-, 256-, and 512-fold diluted immune serum and the 8- to 2 048- fold diluted bulk were tested for titer and median effective concentration (EC₅₀). Results　The titer results of immune serum determined by the method in Chinese pharmacopoeia and the PBMC based flow cytometry method were all 1:512. The titer of batch 001 immune serum determined by the Jurkat cell based flow cytometry method was 1:512, while the titers of batch 002 and 003 were 1∶256. The titers of three batches of bulk determined by the three methods were comparable to the positive control Grafalon, and the titer trends detected by different methods were basically consistent.Conclusion　The trend of the three methods for determining the titers of immune serum and bulk is basically consistent, but the flow cytometry (PBMC and Jurkat) method is more convenient to operate and can quantitatively measure the EC₅₀ value.

Objective　To evaluate the stability of the self-made aluminum phosphate adjuvant under different storage conditions so as to provide supportive data for the storage and expiration date of the adjuvant.Methods　Three consecutive batches of industrial-scale self-made aluminum phosphate adjuvant were selected and stored at (5±3) ℃ and (25±2) ℃, respectively. The main evaluation indices of aluminum phosphate adjuvant (appearance, pH value, aluminum content, adsorption rate, sodium chloride content, analysis of sedimentation supernatant, sterility, bacterial endotoxin, arsenic salt, iron salt, sulfate salt, ammonium salt, heavy metals, phosphorus to aluminum molar ratio, zero charge point, particle size and distribution) were investigated by sampling at regular intervals.Results　The appearance and sterility of 3 batches of aluminum phosphate adjuvant were qualified after stored at two temperatures for 15 months. The pH value was 5.4-6.2, the adsorption rate was 93%-100%, the aluminum content was 2.77-3.36 mg/mL, the sodium chloride content was 7.90-8.60 g/L, and the sedimentation supernatant analysis ranged from 12 to 15 mL .The bacterial endotoxin ranged from 0.5 to 3.0 endotoxin unit/mg aluminum, and the molar ratio of phosphorus to aluminum ranged from 0.82 to 1.13. The particle size corresponding to the cumulative distribution reaching 10%, 50% and 90% were 1.75-2.53, 2.87-3.98 and 4.94-6.90 μm, respectively. The zero charge point was 5.11-5.47, with the residual impurity of arsenic salt ≤0.001‰, iron salt ≤0.015‰, sulfate salt ≤0.5%, ammonium salt ≤0.005% and heavy metal ≤0.002%. All main evaluation indices of the adjuvant were stable and in line with the quality standard.Conclusion　Three batches of self-made aluminum phosphate adjuvant show good stability after being stored at (5±3) ℃ and (25±2) ℃ for 15 months, which provides a basis for the formulation of storage conditions and efficacy of aluminum phosphate adjuvant.

Objective　To evaluate the immunogenicity of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza virus combined vaccine formulated with 4 adjuvants Al(OH)₃, MF59, AS03 and QS21 in mice.Methods　Adjuvants Al(OH)₃, MF59, AS03 and QS21 were used respectively to prepare SARS-CoV-2 (inactivated) and tetravalent influenza virus (split) combined vaccine. BALB/c mice were immunized by intraperitoneal injection on day 0 (D0) and D14, respectively. Blood samples were collected on D14 and D28 to detect antibody titers against SARS-CoV-2 and hemagglutination inhibition titers of influenza. Spleen lymphocytes were isolated on D28 to detect cellular immune responses to both SARS-CoV-2 and influenza virus.Results　SARS-CoV-2 and influenza virus combined vaccine with different adjuvants induced both antigen-specific antibody responses and cellular immune responses in mice. At D28 post-initial immunization, MF59 adjuvant group induced high levels of SARS-CoV-2 binding antibodies and neutralizing antibodies, with geometric mean titers (GMTs) of 89 144 and 5 418, respectively, and MF59 group also induced high levels of hemagglutination-inhibiting antibodies against the quadrivalent influenza virus strains (H1N1, H3N2, BV, BY), with GMTs of 4 457, 5 120, 1 470 and 5 881, respectively. Both the MF59 and AS03 groups induced robust Th1-type (IFN-γ, IL-2) cellular immune responses against SARS-CoV-2, with spot forming units (SFUs) statistically significantly higher than those of Mock group （IFN-γ： H=16.69，P＜0.01； IL-2： H=15.21， P＜0.05）. The AS03 group induced a strong Th1-type (IFN-γ, IL-2) cellular immune response against the quadrivalent influenza virus strains (H1N1, H3N2, BV, BY), with SFUs statistically significantly higher than those of Mock group （IFN-γ： H=12.93, 12.17, 11.82, 13.61, P＜0.05； IL-2： H=12.24, 12.42, 11.72, 12.43， P＜0.05）.Conclusion　The immunogenicity as well as specific antibody and cellular immune responses of SARS-CoV-2 and influenza virus combined vaccine with different adjuvant formulations are different, indicating the importance of adjuvants in the development of combined vaccine formulations.

Objective　To establish and validate a colloidal gold immunochromatographic assay for the rapid and correct detection of monkeypox virus (MPXV).Methods　The labelling conditions of 7 anti-MPXV A35R monoclonal antibodies were optimized by NaCl precipitation assay.The antibody pairing assay was used to determine the gold standard antibody and the capture antibody, and to optimize the spotting concentration of the antibody detection line (T line) and the quality control line (C line), so as to establish the colloidal gold immunochromatographic detection method. The specificity, limit of detection, stability, repeatability, the hook effect, anti-interference performance of the method were verified.Results　The optimal antibody labelling amounts for 7 anti-MPXV A35R monoclonal antibodies 8D7, 10D2, 2F5, 10G4, 7G5, 10E4 and 2D7 were 0.96, 0.48, 0.96, 0.96, 0.96, 0.96, 0.48 μl, respectively, and the combination of the gold-labelled antibody 2D7 and the test antibody 8D7 was the best pair of antibodies. The optimal concentrations for the T and C line spot samples were 1.00 and 0.50 mg/ml, respectively. Colloidal gold test strips could detect MPXV but not other viruses; the minimum detection limits of MPXV A35R protein and inactivated MPXV solution were 2.34×10^-5 mg/ml and 1.00×10⁵ TCID₅₀/ml, respectively; the colour development of the negative samples was consistent with that of the positive samples; the colour development of 20 test strips was consistent; the concentrations of MPXV A35R protein and inactivated MPXV solution were proportional to the degree of colour development; the test strips were not interfered by interfering substances.Conclusion　A colloidal gold immunochromatographic assay is successfully developed for the rapid detection of MPXV, which has good specificity, detection range, stability, reproducibility and anti-interference performance without hook effect.

Objective　To establish and validate a method for the detection of dimethyl sulfoxide (DMSO) in pneumococcal polysaccharide conjugate bulk.Methods　The mobile phase, column temperature and flow rate in the reversed phase high performance liquid chromatography(RP-HPLC) were optimized to establish the optimal detection method. The method was validated in terms of specificity, linearity, accuracy, precision and stability.Results　The RP-HPLC method was optimized with 10% acetonitrile as the mobile phase at a flow rate of 0.6 ml/min and a column temperature of 35 ℃. The relative errors of this method for the detection of DMSO content in the test and control groups of the interfering components ranged from -2.6% to 2.1%, and the resolutions were all ≥1.5; the concentrations showed good linearity with the peak areas, and the regression coefficient of determination was ≥0.990 0; the biases of different concentration levels ranged from -5.31% to -0.89%, and the upper limit of bias in 95% was≤8.0%; the precision of different concentrations ranged from 0.70% to 4.45%, and the upper limit of confidence of the precision in 95% was ＜8.0%. The evaluation of the comprehensive ability of the method showed that the total method variation was ≤7.0% in the range of 4-50 μg/ml. Both the 95% prediction interval and the 95%/90% tolerance interval were in the range of 80%-120%. The relative standard deviation of the mean peak area of the test solution within 20 h were ≤2.0%.Conclusion　The method for the determination of DMSO in pneumococcal polysaccharide conjugate bulk is successfully established, which shows good specificity, linearity, accuracy, precision, stability and high comprehensive evaluation ability.

Objective　To evaluate the feasibility of recombinant trypsin as a substitute for animal-derived trypsin in production of live attenuated mumps and live attenuated measles vaccines.Methods　The required concentration of recombinant trypsin was calculated based on the enzymatic activity of animal-derived trypsin and 0.10-0.25 mg/ml was selected to treat chicken embryo tissues and produce two vaccines. Animal-derived trypsin was used as control. The total number of viable cells, cell adherence rate, and virus titers of the virus harvests were compared and statistically processed.Results　Compared with control group, the tissues of chicken embryos digested with recombinant trypsin appeared more delicate and the cells were easily dispersed. The total numbers of viable cells from each batch of chicken embryos after dispersion were 1.19×10¹⁰ and 1.63×10⁹ for 0.20 mg/ml recombinant trypsin groups in production of mumps and measles vaccines, 9.31×10⁹ and 1.27×10⁹ for control group, respectively, while the cell adherence rates were 63.01% and 82.64% for 0.2 mg/ml groups, 53.90% and 75.99% for control groups. Virus titers of the virus harvests were all within the normal range.Conclusion　Recombinant trypsin at 0.20 mg/ml can be used as a substitute for animal-derived trypsin in the production of live attenuated mumps and measles vaccines.

Objective　To validate the Lowry method 2 for the determination of pertussis toxin (PT) , filamentous hamagglutinin (FHA)，and pertactin (Prn) contents in pertussis antigen. Methods　PT, FHA, and Prn national standards were diluted and the standard curve was established in the concentration range of 0-50 μg/ml. PT, FHA, and Prn refined solutions were added with national standards to prepare standard samples, and were tested for protein contents by Lowry method 2. The specificity was tested for 400 mmol/L PBS used in bulk production. Robustness was validated by testing samples 10, 20, and 30 min after primary test. Results　The recoveries ranged from 91.1% to 110.7%, the accuracy relative standard deviations (RSDs) ranged from 1.8% to 9.2%, and the intermediate precision RSDs ranged from 2.6% to 6.2%. The 400 mmol/L PBS had no interference on the determination of samples. Test results were reliable within 30 min. Conclusion　The Lowry method 2 has good linearity, accuracy, precision, specificity and robustness, and can be used for the determination of PT, FHA, and Prn contents in pertussis antigen.

Objective　To investigate the applicability of cytopathic inhibition assay for the detection of human cytomegalovirus neutralizing antibody (CMV-NA) potency in plasma and human cytomegalovirus immunoglobulin for intravenous injection (CMV-IVIG).Methods　Cytopathic inhibition assay for detection of CMV-NA potency was established with cytomegalovirus immunoglobulin national standard substance. The repeatability, intermediate precision, specificity, accuracy, durability and reproducibility of cytopathic inhibition assay were investigated with CMV-NA plasma and CMV-IVIG.Results　The relative standard deviations of the repeatability validation of CMV-NA potency for plasma and CMV-IVIG were 12% and 19%, respectively. The CMV-NA potency of plasma and CMV-IVIG were detected 6 times by 2 experimentalists, with F values of 0.84 and 0.03, respectively, and P > 0.05. With 6 repetitions of plasma and CMV-IVIG for CMV-NA potency tested on different dates, the F values were 0.89 and 0.55, respectively, with P > 0.05. Plasma, CMV-IVIG and national standard substance were free of cytopathy. The recovery rates of both plasma and CMV-IVIG labeled samples were within 100% ± 40%. The CMV-NA potencies of plasma and CMV-IVIG were tested at various neutralization time, and the F values were 0.48 and 0.04, respectively, with P > 0.05. The CMV-NA potencies of plasma and CMV-IVIG were detected in different viral titer ranges with F values of 0.49 and 0.11, respectively, and P >0.05. The CMV-NA potency of CMV-IVIG was detected by 2 laboratories with a t value of 0.69 and P >0.05.Conclusion　The cytopathic inhibition assay for CMV-NA potency has good repeatability, intermediate precision, specificity, accuracy, durability and reproducibility.

Objective　To establish and verify a fluorescence staining method for the residual DNA content in recombinant hepatitis E vaccine (HepEV).Methods　λDNA was used as the standard to establish the standard curve, and purified recombinant HepEV bulk was used as the sample to verify the linearity, accuracy and precision.Results　The method had good linearity in the range of 1.250-80.000 ng/ml, and the coefficient of determination was greater than 0.99. In the accuracy verification, the recovery rate of DNA residue was between 90% and 110%, and the relative standard deviation (RSD) of the recovery rate was less than 15%. At the same time, the RSDs of repeatability verification and intermediate precision verification were 6.62% and 10.30%, respectively, both less than 15%.Conclusion　This method is rapid, sensitive and accurate, and can detect the residual host DNA content in recombinant HepEV.

Objective　To develop and apply a potentiometric titration method for determination of sodium chloride content in biological products. Methods　With sodium chloride primary standard as control, the content of sodium chloride in sample was determined by potentiometric titration with silver nitrate titrant.The same samples were determined and compared by different methods. Results　The concentration of sodium chloride in the range of 6-15 mg/ml showed a good linear relationship with the volume of silver nitrate titrant consumed. The coefficient of determination for the regression equation was 0.999 9. The average recovery rates of sodium chloride in meningococcal polysaccharide vaccine culture medium, tetanus antitoxin stock solution, Haemophilus influenzae type b conjugate ,aluminum hydroxide adjuvant for vaccine were 100.0%, 99.8%, 100.2% and 100.3%,respectively, which showed that the potentiometric titration method had good accuracy. The maximum relative standard deviation of the six determinations of the same batch of samples was 0.7%, indicating that the method had good repeatability. The test samples were placed for 5 and 30 min, respectively, after dilution and then tested, which had no obvious effect on the results, indicating that the method had good durability. Statistical analysis of the results obtained by potentiometric titration method and the Volhard method adopted in Chinese pharmacopoeia 2020 edition showed that there was no statistically significant difference between the results of the two methods (t values were -2.12—2.50，P>0.05). Conclusions　The method of potentiometric titration for the determination of sodium chloride content in biological products is established. The method has good linear range, good repeatability and accurate quantification, and is suitable for the determination of sodium chloride content in biological products.

Vaccine is a special drug containing bioactive substances, and the storage and transportation of vaccine have certain cold chain requirements on temperature. After the Shandong vaccine case, a series of laws and regulations have been issued to further strengthen the management of vaccine production, circulation and vaccination. In this paper, the laws and regulations related to vaccine circulation management are systematically combed and analyzed in chronological order, aiming at clarifying the context of laws and regulations for vaccine production, distribution, storage enterprises and vaccination agencies, and providing legal and regulatory basis for them to better carry out vaccine circulation management.

Objective　To develop a chemiluminescence microparticle immunoassay(CMIA) detection reagent for hepatitis C virus core antigen (HCV-cAg) and preliminarily evaluate its performance. Methods　Two anti-HCV-cAg monoclonal antibodies (Mabs) were pre-coated with magnetic particles, and the other two anti-HCV-cAg Mabs were labeled with acridine ester to prepare a CMIA detection reagent for double-antibody sandwich magnetic particles. Using CMIA and ELISA kit,210 interference samples, 1 158 normal human samples, 140 HCV-RNA positive samples, and 2 sets of serum conversion disk samples were detected simultaneously to identify the specificity and sensitivity of the developed reagent. Results　The specificity of CMIA detection reagent was 99.74%, and the specificity of ELISA kit was 99.57%. The negative rates of both methods for detecting interference samples were 100%. The sensitivity of CMIA detection reagent was 77.86%, the sensitivity of ELISA kit was 62.14%, and the CMIA detection reagent detected positive serum conversion disk samples earlier than ELISA kit. Conclusion　HCV-cAg CMIA detection reagent is successfully prepared with good specificity and high sensitivity, and is suitable for early screening and clinical auxiliary diagnosis of HCV.

Drug marketing authorization holder takes the main responsibility for drug quality and safety. This paper reviews the implementation of the drug marketing authorization holder system at home and abroad, studies the regulations on the supervision and administration of drug marketing authorization holder implementing the main responsibility of drug quality and safety, which was implemented on March 1, 2023, and further comments on the responsibilities and requirements of drug marketing authorization holder in carrying out drug quality management.

Objective　To benchmark the quality control indicators of the homemade rabbit derived type 5 pneumococcal specific serum by comparing with the commercial reference products, in order to detect the process samples in the production of 23-valent pneumococcus polysaccharide vaccine.Methods　The non-competitive turbidity mode of the dual path rate turbidity immunoassay system was used for detection. The working concentration, linearity, crossover, comparability with commercially available serum, detection difference both between and within laboratories， stability under working concentration of type 5 sera were investigated in turn.Results　The maximum dilution multiples of the working concentrations for the 4 batches of home made type 5 sera were 75, 20, 30 and 8 times, respectively. Under the corresponding working concentrations, the polysaccharide concentration showed a good linear relationship with the response value in the range of 0.625-4.000 μg/ml, and the linear correlation coefficients were all > 0.985. There was no crossover between 4 batches of type 5 sera reference samples and the other 22 types, and the specificity was good. In the comparability test with commercially available serum, the coefficient of variation (CV) of type 5 polysaccharide content in 3 different manufacturers' marketed vaccines was less than 5% for all 5 batches of sera including commercially available serum. There was no apparent difference in the detection results between the 4 batches of type 5 sera and commercially available serum as the weight-average molecular weight decreased. The CV values of between- and within-department outdoor testing results were less than 10%. Four batches of type 5 sera with working concentration maintained good stability for at least one month under 2—8 ℃.Conclusion　The homemade rabbit derived type 5 pneumococcal specific serum has high potency, good linearity,no crossover, comparability to commercially available serum， and good stability at working concentrations， and can be used for polysaccharide content detection of type 5 pneumococcus polysaccharide vaccine.

Genital tract infections with Chlamydia trachomatis (CT) can lead to serious complications including pelvic inflammation, tubal inflammation, ectopic pregnancy, and infertility in females. However, the pathogenesis of CT remains unclear. The deficiency of CT plasmid gene protein 3 (Pgp3) reduces the pathogenicity of CT infections in the upper genital tract and prevents the induction of lesions in the upper genital tubes. These suggest that Pgp3 play a significant role in CT pathogenesis. Pgp3 maintains CT survival and promotes its infection uptake through a variety of processes including inhibition of host cell apoptosis, improvement of host cell survival, and inhibition of human antimicrobial peptides. Pgp3 also establishes persistent infection of CT by promoting the transfer and colonization of CT into the gastrointestinal tract. Furthermore, Pgp3 promotes the development of inflammation by inducing the generation of inflammatory factors. Therefore, Pgp3 is an important virulence factor responsible in CT infection. This article summarizes the research progress on the pathogenic mechanism of CT by Pgp3.

Objective　To establish an environmental bacterial bank in clean area by monitoring, collection and identification of environmental bacteria in pilot clean area of research and development.Methods　Static and dynamic environmental monitoring was carried out in the pilot clean area of research and development, and the collected environmental bacteria (settling microbe, airborne microbe and surface microbe) were identified by morphological observation, staining microscopy and genotypic microbial identification (bacterial 16S ribosomal RNA sequencing analysis).Results　A total of 89 strains of environmental bacteria were isolated, belonging to 16 genera, among which Bacillus sp. accounted for 29%, Staphylococcus sp. accounted for 27% and Micrococcus sp. accounted for 12%.Conclusion　The environmental bacterial bank in pilot clean area of research and development is initially established, which provides technical support for microbial contamination traceability.

Infants and young children are at high risk of influenza, and vaccination is the most cost-effective way to prevent influenza. Currently, trivalent and quadrivalent influenza vaccines are available for children in China. The dosage forms include half dose (7.5 μg hemagglutinin/0.25 ml) and full dose (15 μg hemagglutinin/0.5 ml). Studies have found that the age of infants and young children is positively correlated with the level of immune response, and the immune effect of influenza vaccine for infants and young children can be improved by increasing the antigen content. Full-dose quadrivalent influenza vaccine for infants and young children has lately come to market in the Chinese mainland. This article reviews the epidemic status of influenza virus and the application progress of full-dose quadrivalent influenza vaccine for Chinese infants and young children from the aspects of disease burden of influenza, vaccination status, protective effect, immunogenicity and safety of influenza vaccine, and vaccination intention.

Objective　To establish a rapid method for the detection of mycoplasma using real-time quantitative PCR (qPCR) technology.Methods　Eight mycoplasma species available for validation under European Pharmacopoeia（10.0 edition） 2.6.7 were selected. Specific primers were designed based on mycoplasma 16S RNA sequence. A qPCR method for the detection of mycoplasma was established and validated for specificity, sensitivity and reproducibility.Results　Suitable primers for the detection of mycoplasma by qPCR were successfully screened. Eight mycoplasma species showed a cycle threshold (CT) between 20-30 at a concentration of 10⁵-10⁶ CFU/mL, and there was no CT for the bacterial control at same concentration. Except for Mycoplasma sperminum, which could be detected at a concentration of 10-100 CFU/mL, the other 7 mycoplasma species could be detected at a concentration of 10 CFU/mL. CTs were between 35-40 at 10 CFU/mL. In specificity experiment that repeated 3 times, the control bacteria were all undetected, CTs of 8 species within 10⁵-10⁶ CFU/mL were between 20-30, and for each species, the coefficient of variation (CV) for 3 times was less than 10%. In sensitivity experiment that repeated 3 times, the CTs of each species with different dilution concentrations were between 20-40 in each experiment, and the CVs were less than 10%.Conclusion　The established method has good specificity, sensitivity and reproducibility, and can support timely control of possible mycoplasma contamination risks in biologics.

Objective　To investigate the trends in HBV infection and serological dynamics in low birth weight infants born to HBV surface antigen (HBsAg)-positive mothers, and to analyze HBV infection and the effectiveness of immunization in this very special group of children.Methods　Low birth weight infants born to HBsAg-positive mothers in 24 hospitals or obstetrics departments in Lu'an City from 2016 to 2021 were selected, categorized and counted by name, gender, weight, preterm delivery, hepatitis B vaccine（HepB） timely vaccination, vaccination dose, etc. The concentrations of HBsAg and hepatitis B surface antibody (HBsAb) were quantified by magnetic particulate chemiluminescence, and the antibody positivity rate and geometric mean concentration (GMC) of antibodies were analyzed and compared.Results　A total of 172 low birth weight infants were included in this study, of which 121 completed blood sample collection, with a mean age of (3.7±1.7) years, a median weight of 2.30 kg, a proportion of 58.68% of preterm infants, a proportion of 75.21% of infants who received the first dose of HepB timely The number of subjects who completed HepB primary and booster immunizations were 117 and 4, respectively, and the prevalence of HBV infection was 5.79%. Among the uninfected subjects, there was a statistically significant difference between HBsAb GMC of the booster immunization group and the primary immunization group, with Z=-2.17, P=0.030. Among the uninfected and primarily vaccinated subjects,timely vaccination preterm delivery, and birth weight diden´t effect GMC. Among the HBsAb positive subjects, no statistically significant difference in the positivity rate was seen in the primary immunization group compared to the booster immunization group (χ²=2.45, P=0.118); but there was a statistically significant difference between the two groups in GMC (Z=-2.25, P=0.024). In primarily vaccinated and uninfected subjects, there was an overall trend of decreasing HBsAb positivity and GMC after vaccination.Conclusion　In Lu'an, the HBV infection rate of low birth weight infants born to HBsAg-positive mothers is high, the HepB vaccination rate is low, and the total levels of HBsAg and HBsAb are low, so it is necessary to strengthen the implementation of 0-1-2-7 months HepB immunization program for low birth weight infants.