ObjectiveThis study aims to investigate the dynamic changes in general body parameters, blood glucose, and blood lipid profiles in obese cynomolgus monkeys, exploring the correlations among these parameters and providing a reference for research on the obese cynomolgus monkey model. Methods30 normal male cynomolgus monkeys aged 5 - 17 years old (with body mass index < 35 kg/m² and glycated hemoglobin content < 4.50%) and 99 spontaneously obese male cynomolgus monkeys (with body mass index ≥35 kg/m² and glycated hemoglobin content < 4.50%) were selected. Over a period of three years, their abdominal circumference, skinfold thickness, body weight, body mass index, fasting blood glucose, glycated hemoglobin, and four blood lipid indicators were monitored. The correlations between each indicator were analyzed using repeated measurement ANOVA, simple linear regression, and multiple linear regression correlation analysis method. Results Compared to the control group, the obese group exhibited significantly higher levels of abdominal circumference, skinfold thickness, body weight, body mass index, and triglyceride (P＜0.05). In the control group, skinfold thickness increased annually, while other indicators remained stable. Compared with the first year, the obese group showed significantly increased abdominal circumference, skinfold thickness, body weight, body mass index, triglyceride, and fasting blood glucose in the second year(P＜0.05), with this increasing trend persisting in the third year (P＜0.05). In the control group, the obesity incidence rates in the second and third years were 16.67% and 23.33%, respectively, while the prevalence of diabetes remained at 16.67%. In the obese group, the diabetes incidence rates were 29.29% and 44.44% in years 2 and 3, respectively. Among the 11-13 year age group, the incidence rates were 36.36% and 44.68%, while for the group older than 13 years, the rates were 28.13% and 51.35%. Correlation analysis revealed significant associations (P＜0.05) between fasting blood glucose and age, abdominal circumference, skinfold thickness, body weight, and triglyceride in the diabetic monkeys. Conclusion Long-term obesity can lead to the increases in general physical indicators and fasting blood glucose levels in cynomolgus monkeys, and an increase in the incidence of diabetes. In diabetic cynomolgus monkeys caused by obesity, there is a high correlation between their fasting blood glucose and age, weight, abdominal circumference, skinfold thickness, and triglyceride levels, which is of some significance for predicting the occurrence of spontaneous diabetes.

As the need for antibody production rises, there is an urgent need to lower the costs and enhance the efficiency of the separation process. Currently, the chromatographic media used for antibody separation and purification often focus on individual properties of antibodies, such as affinity, hydrophobicity, and charge, leading to issues like low purification efficiency or inadequate adsorption capacity. To address this, an electrostatically coupled polypeptide affinity medium (FD7-3, 5-diaminobenzoic acid n-sepharose, FD7-DA-Sepharose) was developed for rapid purification of antibodies from cell culture supernatant. This medium utilized 3, 5-diaminobenzoic acid as a spacer to attach the heptapeptide-affinity ligand (FYEILHD, FD7) to agarose microspheres. Antibodies could be adsorbed through charge interactions with the carboxyl functional group of the FD7-DA-Sepharose spacer, while FD7 enhanced electrostatic coupling and affinity adsorption through synergistic effects, significantly increasing the adsorption capacity while maintaining the affinity and specificity. The influences of pH and ionic strength on adsorption capacity were investigated with human immunoglobulin as a model protein. The static adsorption capacity (Q_m) of FD7-DA-Sepharose in the solution of pH 6.0 reached 67.73 mg/mL, representing a 52.68% increase compared with that (44.36 mg/mL) of the commercial Protein A affinity medium. Furthermore, the elution conditions for FD7-DA- Sepharose were mild (20 mmol/L PB, 0.5 mol/L NaCl, pH 6.0), in contrast to the harsh acidic elution (pH 2.7-3.6) typically associated with Protein A, which can damage antibody integrity. The FD7-DA-Sepharose medium was then employed to purify antibodies from cell culture supernatant, achieving the yield of 94.8% and the purity of 98.4%. The secondary structure of the purified antibody was determined by circular dichroism spectroscopy. The results demonstrated that FD7-DA-Sepharose enabled efficient purification of antibodies from cell culture supernatant, which provided a cost-effective solution (approximately one-third the price of commercial Protein A affinity medium) with gentle elution conditions that preserve the natural conformation of antibodies. This approach paves a novel, economical, and efficient way for the separation and purification of antibodies from cell culture supernatant.
Chromatography, Affinity/methods* ; Static Electricity ; Humans ; Sepharose/analogs & derivatives* ; Peptides/chemistry* ; Adsorption ; Antibodies/isolation & purification*

Objective To investigate the application value of procalcitonin(PCT),neutrophil-to-lym-phocyte ratio(NLR)and C-reactive protein to albumin ratio(CAR)in the short-term prognosis of the pa-tients with severe community acquired pneumonia(SCAP).Methods A total of 225 patients with communi-ty-acquired pneumonia(CAP)treated in this hospital from July 2021 to December 2022 were selected as the study subjects and divided into the SCAP group and non-SCAP group.Then the patients with SCAP were di-vided into the survival group and death group according to their survival state in 28 d.The general data and re-lated laboratory indexes were collected.NLR and CAR were calculated.The binary logistic regression was a-dopted to analyze the risk factors of death on 28 d for the SCAP patients.The receiver operating characteristic(ROC)curve was drawn and the area under the curve(AUC)was calculated to evaluate the predictive value of relevant indicators.Results The age,levels of PCT,NLR,CAR,WBC and NEU,and the proportion of cor-onary heart disease in the SCAP group were higher than those in the non-SCAP group,while the levels of LYM and ALB were lower than those in the non-SCAP group,and the differences were statistically significant(P＜0.05).The age,levels of PCT,NLR,CAR,WBC and NEU,and the proportion of COPD in the death group were higher than those in the survival group,while the levels of LYM and ALB were lower than those in the survival group,and the differences were statistically significant(P＜0.05).The binary logistic regres-sion analysis showed that the age(OR=1.069,95％CI:1.020-1.120),chronic obstructive pulmonary dis-ease(OR=5.633,95％CI:2.019-15.712),PCT(OR=1.045,95％CI:1.002-1.090)and CAR(OR=2.170,95％CI:1.616-2.915)were the independent influencing factor for the death on 28 d in the patients with SCAP.The ROC curve analysis indicated that AUC,sensitivity and specificity of PCT,NLR and CAR combined detection were superior to those of detection alone,having good predictive value.Conclusion PCT,NLR and CAR have a certain predictive value for death within 28 d in the patients with SCAP.The combined detection has better predictive efficiency.

Objective: To investigate the value of metagene next⁃generation sequencing ( mNGS) in the detection of pathogens in patients with pulmonary infection. Methods: A retrospective analysis was performed on clinical data from 434 patients with pulmonary infections admitted over the past four years. Based on the presence of underlying comorbidities , patients were divided into underlying disease group (n = 262) and non⁃underlying disease group (n = 172) . Pathogen detection was conducted using both mNGS and conventional tests. Clinical and laboratory parameters , radiographic findings , and pathogen detection results were systematically analyzed. The diagnostic performance of the two methods in identifying causative pathogens of pulmonary infections was compared. Results: The positive rate of mNGS in 434 patients was higher than that of conventional tests , and the difference was statisti⁃cally significant (P < 0. 05) . The efficacy of mNGS in detecting bacteria and viruses was significantly higher than that of conventional tests , and the difference was statistically significant (P < 0. 05) . Although the fungal detection rate of mNGS was higher than that of conventional tests , the difference was not statistically significant. Among them , the detection rates of Mycobacterium tuberculosis , Mycoplasma pneumoniae , Haemophilus influenzae , Strepto⁃ coccus pneumoniae , Streptococcus constellation , Staphylococcus aureus and Aspergillus fumigatus were significantly higher than those of conventional tests , and the difference was statistically significant (P < 0. 05) . Subgroup analy- sis showed that the proportion of males , hospital stay , smoking prevalence and average age in the underlying dis- ease group were higher than those in the non-underlying disease group , and the difference was statistically signifi- cant (P < 0. 05) , while there were no significant differences in antibiotic use and endotracheal intubation rate be- tween the two groups. The most common pathogens detected by mNGS in the underlying disease group were Myco⁃ bacterium tuberculosis , Haemophilus influenzae , Streptococcus pneumoniae , Pseudomonas aeruginosa , human herpes⁃ virus type 4 and Aspergillus fumigatus , while the most common pathogens in the non-underlying disease group were Mycobacterium tuberculosis , Haemophilus influenzae , Streptococcus pneumoniae , Mycoplasma pneumoniae and Kleb⁃ siella pneumoniae. The positive rate of mNGS in the two groups was significantly higher than that of conventional tests , and the difference was statistically significant (P < 0. 05) , while the difference in the positive rate of mNGS between the two groups was not statistically significant. Conclusion mNGS has significant advantages over con- ventional tests of pathogen in lung infection , and is less affected by underlying diseases , which can provide an etio- logical basis for lung infection.

Threatened abortion is a common disease of obstetrics and gynecology and one of the diseases responding specifically to traditional Chinese medicine （TCM）. The China Association of Chinese Medicine organized experts in TCM obstetrics and gynecology， Western medicine obstetrics and gynecology， and pharmacology to deeply discuss the advantages of TCM and integrated Chinese and Western medicine treatment as well as the medication plans for threatened abortion. After discussion， the experts concluded that chromosome， endocrine， and immune abnormalities were the key factors for the occurrence of threatened abortion， and the Qi and blood disorders in thoroughfare and conception vessels were the core pathogenesis. In the treatment of threatened abortion， TCM has advantages in preventing miscarriages， alleviating clinical symptoms and TCM syndromes， relieving anxiety， regulating reproductive endocrine and immune abnormalities， personalized and diversified treatment， enhancing efficiency and reducing toxicity， and preventing the disease before occurrence. The difficulty in diagnosis and treatment of threatened abortion with traditional Chinese and Western medicine lies in identifying the predictors of abortion caused by maternal factors and the treatment of thrombophilia. Recurrent abortion is the breakthrough point of treatment with integrated traditional Chinese and Western medicine. It is urgent to carry out high-quality evidence-based medicine research in the future to improve the modern diagnosis and treatment of threatened abortion with TCM.

We developed a method for accurate quantification of the intact virus particles in inactivated avian influenza virus feedstocks. To address the problem of impurities interference in the detection of inactivated avian influenza virus feedstocks by direct high performance size exclusion chromatography (HPSEC), we firstly investigated polyethylene glycol (PEG) precipitation and ion exchange chromatography (IEC) for H5N8 antigen purification. Under the optimized conditions, the removal rate of impurity was 86.87% in IEC using DEAE FF, and the viral hemagglutination recovery was 100%. HPSEC was used to analyze the pretreated samples. The peak of 8.5-10.0 min, which was the characteristic adsorption of intact virus, was analyzed by SDS-PAGE and dynamic light scattering. It was almost free of impurities and the particle size was uniform with an average particle size of 127.7 nm. After adding antibody to the IEC pretreated samples for HPSEC detection, the characteristic peak disappeared, indicating that IEC pretreatment effectively removed the impurities. By coupling HPSEC with multi-angle laser scattering technique (MALLS), the amount of intact virus particles in the sample could be accurately quantified with a good linear relationship between the number of virus particles and the chromatographic peak area (R²=0.997). The established IEC pretreatment-HPSEC-MALLS assay was applied to accurate detection of the number of intact virus particles in viral feedstocks of different subtypes (H7N9), different batches and different concentrations, all with good applicability and reproducibility, Relative standard deviation < 5%, n=3.
Animals ; Reproducibility of Results ; Influenza A Virus, H7N9 Subtype ; Influenza in Birds ; Chromatography, Gel ; Virion ; Lasers

Objective To explore the safety and efficiency of sequential operation of Tirelizumab combined with chemotherapy in neoadjuvant therapy for locally advanced esophageal cancer.Methods A retrospective analysis was conducted on 60 patients with locally advanced esophageal cancer admitted to Renmin Hospital of Wuhan University from August 2018 to June 2022.The immunotherapy combined with chemotherapy sequential surgery patients were selected as the observation group(29 cases)and the chemotherapy sequential surgery patients were selected as the control group(31 cases)according to the treatment method.The study aimed to analyze whether there were differences in efficacy and safety between the two groups.Results There are 28 patients with R0 resection in the observation group,and 14 case reached ORR.In the control group,29 cases were resected with R0,and 7 cases reached ORR.The proportion of ORR patients in the observation group was significantly higher than that in the control group,which was statistically significant(P＜0.05).There were 9 patients with pCR in the observation group and 2 patients with pCR in the control group,and the proportion of pCR patients in the two groups was significantly different(P＜0.05).There were no significant differences between the two groups for preoperative and postoperative adverse events.Conclusion Immune checkpoint inhibitors combined with chemotherapy sequential surgery are safe and reliable in patients with locally advanced esophageal cancer,with significant short-term efficacy,and long-term efficacy remains to be observed.

Objective To summarize the clinical experience in the treatment of postoperative upper gastrointestinal cancer of esophageal cancer.Methods The clinical data of 16 patients with postoperative upper gastrointestinal malignancies treated in our hospital from January 2018 to June 2022 were retrospectively analyzed.Results All the 16 patients successfully completed the operation,and no perioperative death occurred.The cumulative length of hospitalization was 18-38 days.After operation,2 cases of pulmonary infection,1 case of respiratory failure,and 1 case of cervical anastomotic fistula were cured after conservative treatment.All patients could eat normally during postoperative follow-up,and no tumor recurrence and metastasis was found.Conclusion For patients with recurrent upper gastrointestinal cancer after esophageal cancer surgery,if the lesion is relatively limited,surgical treatment is reliable and an optional treatment plan.

This paper aims to detect the antigens in porcine circovirus type 2 (PCV2) vaccines by high-performance size-exclusion chromatography (HPSEC) coupled with multi-angle laser light scattering (MALLS). With purified inactivated PCV2 and PCV2 virus-like particles (VLP) as references, two inactivated vaccines (a and b) and two VLP vaccines (c and d) for PCV2 from four manufacturers were analyzed by HPSEC-MALLS after demulsification. The antigen peaks in HPSEC-MALLS were identified by PCV2 antigen test strips, Western blotting and transmission electron microscope (TEM). The repeatability and linearity of the method were investigated. The results showed the virus antigens in the two inactivated vaccines were eluted at about 13.3 min in HPSEC. The molecular weight of these antigens was 2.61×10⁶ (±4.34%) Da and 2.40×10⁶ (±2.51%) Da, respectively, as calculated by MALLS. The antigen peaks of the two VLP vaccines also appeared at 13.3 min and the molecular weight was 2.09×10⁶ (±2.94%) Da and 2.88×10⁶ (±11.85%) Da, respectively, which was close to the theoretical molecular weight of PCV2. Moreover, an antigen peak of VLP vaccine c was observed at 11.4 min and the molecular weight was 4.37×10⁶ (±0.42%) Da. The antigen was verified to be the dimer of VLP by TEM. Vaccine d and purified Cap VLP antigens were tested repeatedly, and the RSD of the peak area (n=3) was all < 1.5%, indicating that the method was repeatable. The purified VLP were diluted in serial and tested for linearity. The result suggested good linear relationship between the peak area of VLP or VLP aggregates and the protein concentration of the sample with R² of 0.999 and 0.997, respectively. Thus, the method met the requirement for quantification and aggregate analysis. This method is accurate and efficient in in vitro quality evaluation and improvement of PCV2 vaccine.
Animals ; Antibodies, Viral ; Capsid Proteins ; Chromatography, Gel ; Circoviridae Infections/prevention & control* ; Circovirus ; Lasers ; Swine ; Vaccines, Inactivated ; Vaccines, Virus-Like Particle ; Viral Vaccines

Immunotherapy is becoming an effective and less invasive strategy that can be applied to the treatment of various malignancies. Lentiviral vectors (LVs) have shown great potential in immunotherapy as they can stably integrate relatively large foreign DNA, and effectively transduce dividing and non-dividing cells. Clinical application needs high quality LVs, and therefore strict quality control of the final products is necessary to ensure their purity, efficacy and safety. The quantitative detection of LVs is among the key parts of product development and quality control. In this paper, the existing methods for quantitative detection of LVs are summarized, including fluorescence activated cell sorter (FACS), P24 enzyme-linked immuno sorbent assay (P24 ELISA), real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), nanoparticle tracking analysis (NTA), tunable resistive pulse sensing(TRPS) and virus counter(VC).Their advantages and disadvantages are listed, and future development and challenges are discussed.
Genetic Vectors/genetics* ; Humans ; Immunotherapy ; Lentivirus/genetics* ; Neoplasms ; Transduction, Genetic