1.Development and verification of an indirect ELISA method for determination of specific IgG antibody against SARS-CoV-2 variant strain in rhesus monkey serum
Chinese Journal of Biologicals 2023;36(11):1341-1346+1352
Objective To develop and verify an indirect ELISA method for determination of specific IgG antibody of rhesus monkey serum against SARS-CoV-2 variant strain. Methods An indirect ELISA method for the determination of specific IgG antibody was developed using inactivated SARS-CoV-2 Beta variant strain inactivated vaccine as coating antigen,and optimized for the coating antigen concentration(1,2 and 4 μg/mL),dilution of serum(1∶800~1∶12 800),blocking solution(PBST containing 1% BSA,2% BSA,1% skim milk,2% skim milk and 1% BSA + 1% skim milk),blocking time(30,60 and 90 min),dilution of secondary antibody(1∶5 000,1∶10 000,1∶15 000 and 1∶20 000),incubation time of serum and secondary antibody(30,60 and 90 min),and TMB reaction time(5,10,15,20,25 and 30 min). 60 negative serum samples of rhesus monkeys were detected by the developed method,and the negative and positive critical values were determined. The sensitivity and precision of the methodology were verified. In addition,the specific IgG antibody and neutralizing antibody against SARS-CoV-2 Beta variant strain in 44 serum samples of rhesus monkey were detected by the developed method and microneutralization method,and the correlation and consistency between the two methods were compared. Results The optimum detection conditions were determined:the coating antigen concentration was 1 μg/mL;the blocking solution was PBST containing 1% skim milk,and the blocking time was 30 min;the serum samples to be tested were diluted to 1∶1 600 and incubated for 90 min,and the secondary antibody was diluted to 1∶10 000 and incubated for 30 min;the color development time of substrate was 25 min. The positive critical value and negative critical value of the method was 0. 093 and 0. 084 respectively,and the detection values between them were judged as suspicious. The dilution of5 positive serum samples that showed positive results was 1∶51 200;the coefficients of variation(CVs)of precision were all less than 15%. There was a strong correlation between IgG antibody titer and neutralizing antibody level in the 44 rhesus monkey serum samples(r = 0. 858 0,P < 0. 000 1);the total coincidence rate of the two methods was 90. 9%,the positive coincidence rate was 93. 6%,and the negative coincidence rate was 84. 6%;the consistency test Kappa value was 0. 783 8(95% CI:0. 586 5~0. 981 0). Conclusion The developed indirect ELISA method for eletermination of specific IgG antibody against SARS-CoV-2 Beta variant strain in rhesus monkey serum has good sensitivity and precision,and has strong consistency with microneutralization method,which can be used for the determination of IgG antibody in rhesus monkey serum.
2.Clinical effect of uterine arterial embolization combined with hysteroscopes in the treatment of cesarean scar pregnancy
Chinese Journal of Postgraduates of Medicine 2012;35(18):10-12
Objective To investigate the clinical effect of uterine arterial embolization combined with hysteroscopes in the treatment of cesarean scar pregnancy(CSP).Methods Seventy CSP patients were divided by willingness of patients and their familiality into treatment group and control group with 35 cases each.The treatment group was treated with uterine arterial embolization combined with hysteroscopes,while the control group was treated with uterine curettage after crug therapy.The intraoperative and postoperative situations of two groups were observed.Results The intraoperative blood loss,erythrocyte suspension transfusion,plasma transfusion,preparation time before curettage,hospital stay and pestoperative serum β- human chorionic gonadotropin in treatment group were all obviously lower than those in control group [(420.7 ± 18.9) ml vs.(1893.9 +39.8) ml,(2.1 + 1.3) U vs.(11.1 ±1.4) U,(400.2 ±23.6) ml vs.(1700.3 ± 31.4) ml,(3.9 ± 2.3) d vs.( 18.8 ± 6.7) d,( 14.32 ± 3.89) d vs.(37.83 ± 8.98) d,(312 ± 54) U/L vs.(532 ± 85) U/L,P< 0.01].There was 1 case with massive hemorhage in treatment group,while 3 cases in control group.Conclusions Uterine artery embolization combined with hysteroscopes is a safe and effective treatment with less trauma for CSP.It is worthy of clinical promotion and application.
3.Short-term effect of open and laparoscopic cholecystectomy surgery on body
Chinese Journal of Postgraduates of Medicine 2014;37(8):49-51
Objective To evaluate the short-term effect of open and laparoscopic cholecystectomy surgery on body.Methods Forty patients with cholecystectomy were divided into open surgery group and laparoscopic surgery group by random digits table method with 20 cases each.The clinical index was compared between two groups.The immunoglobulin G(IgG) and procalcitonin (PCT) before surgery and 4 d after surgery were determined.Results The operative blood loss,anal exhaust time,length of stay was (135.80 ±26.74) ml,(1.6 ±0.9) h,(6.7 ±3.2) d in open surgery group,which was better than that in laparoscopic surgery group [(63.41 ± 15.06) ml,(0.4 ± 0.3) h,(2.8 ± 1.9) d],and there was significant difference (P < 0.05).The incidence of heat was 15%(3/20) in open surgery group,which was higher than that in laparoscopic surgery group (0),and there was significant difference (P <0.05).There was no significant difference in the IgG and PCT between two groups before surgery (P > 0.05).The IgG and PCT at 4 d after surgery was (13.93 ± 1.98) g/L,(2 153.2 ± 1 452.1) mg/L in open surgery group,which was higher than that in laparoscopic surgery group[(9.23 ± 2.87) g/L,(768.0 ± 237.0) mg/L],and there was significant difference (P < 0.05).Condmion Laparoscopic cholecystectomy has smaller short-term effects on body,while the body of the humoral and cellular immune function in mild,conducive to the body's recovery.
4.Effect of Pidotimod on Expressions of IL-2 and IL-10 of Patients with Genital Herpes and Curative Effect Observation
Chenhong ZHANG ; Wenjin ZHANG ; Huiying LIN
Chinese Journal of Pharmacoepidemiology 2007;0(03):-
Objective:To investigate the effect of pidotimod on the expressions of IL-2 and IL-10 of patients with genital herpes(GH) and obverse its curative effect and quality of life of the patients.Mehtods:45 cases of outpatients with GH were selected as experimental group wich were given pidotimod orally 0.8g per day for 3 months.Observe the curative effect and value the quality of life before and 3 months after and 6 months after the treatment respectively.Detect the change of the expressions of IL-2 and IL-10 of the blood plasma of the GH patients.Select 30 healthy people as control group.Results:The recurrence rate of the experimental group during 1-3 month,4-6 month,7-9 month after treatment were 60.0%、42.2%and 22.2%respectively.The scores of quality of life and the total evaluation of quality of life itself (G1) and health condition(G4) at 3 and 6 month after treatment rising significantly than before treatment,and it is more obviously at 6 months after treatment than 3 months after treatment.Before treatment,the level of IL-2 of experimental group is significantly lower than that of control group,while the level of IL-10 of the experimental group is significantly higher than that of control group.3 and 6 months after treatment,the level of IL-2 rising significantly,while the level of IL- 10 reducing significantly.The change is more obviously at 6 months after treatment than 3 month after treatment either in the IL-2 rising or the IL-10 reducing.Conclusion:There is cellular immune function disorder in the GH patients.Pidotimod has a certain curative effect on the GH patients.It can reduce the recurrence rate of GH and improve the quality of life of the patients.Maybe it can enhance the cellular immune function of the body through rising the level of IL-2 and reducing the level of IL-10.
5.Investigation in lower extremiy injuries of new recruits in a Chinese special military force
Wenjin LI ; Lei SUN ; Jun ZHANG
Orthopedic Journal of China 2006;0(12):-
[Objective] To study the features in lower extremiy injuries of new recruits in the Chinese special military force during Basic Training Program,and supply scientific precautionary measures to reduce the military training injury.[Method]A special military force group had been chosen and the circumstance of military injuries in lower extremiy had been observed from Jan.to March in 2006.The population selected for study included all 281 new recruits of the unit.A questionnaire was designed to document all of the correlated information,and all injuried soldiers were examined and inquired about the case history.All data had been classified for analysis.[Result]In all 281 samples,75 cases occurred lower extremiy injuries(up to 26.69%).In all lower extremiy injuries,35 cases were leg injuries(up to 46.67%),17 cases were ankle injuries(up to 22.67%),11 cases were foot injuries(up to 14.67%).In all injuries,20 cases were acute injuries(up to 26.67%),55 cases were excess fatigue injuries(up to 73.33 %).In all injuries,38 cases were bone and joint injuries(up to 50.67%),37 cases were soft tissue injuries(up to 49.33%).The reasons for injuries were training field(l 8 cases),exercise overload(61 cases).In all training programs,64 cases were occured in 3 kilo run.[Conclusion]Lower extremiy injuries are common and frequently occurring illness.The features and methods of injuries are different and complicated.In this study,excess fatigue injuries caused by exercise overload were the most important.After comprehened the occurring rule,it should be emphasized on health propaganda.During basic military training,we should pay attention to soldiers' health condition.Training should be carried out step by step,to reduce the occurrence lower extremiy training injuries.
6.Research of Field Seedling Technique on Seed Germination and Growth of Desmodium styracifolium(Osb.) Merr.
Fenglian CHEN ; Wenjin ZHANG ; Honghua XU
Journal of Guangzhou University of Traditional Chinese Medicine 2001;0(03):-
Objective To observe the influence of sowing time and planting density on seed germination percentage and growth of Desmodium styracifolium(Osb.) Merr..Methods After quality inspection of the seed,the seeds planting test for Desmodium styracifolium(Osb.) Merr.was carried out from March to October,and the influences of sowing time and planting density on the seed germination percentage and growth were observed.Results On the planting base of Pingyuan county in Guangdong province,the seeds of Desmodium styracifolium(Osb.) Merr.planted from March to October had a high germination percentage and the seedling grew well.With the increase of planting density,the seed germination percentage decreased,but was up to 80% when the planting density was 0.5~1.0 g /m2.Conclusion The optimum planting time for Desmodium styracifolium(Osb.) Merr.is in autumn and the optimum planting density is 0.5~1.0 g /m2.
7.Effect of ginseng stem-leaf saponin on mice's brain aging
Quhua ZHANG ; Wenjin SUN ; Baozhao JU ;
Chinese Traditional Patent Medicine 1992;0(03):-
Objective: To study ginseng stem leaf saponin's effect on brain aging. Methods: The natural aging mice were adopted, divided randomly groups by weight and fed with ginseng stem leaf saponiu for three months, learning and memory ability of mice, the content of MDA and the action of MAO B of brain tissue were determined. The mice was fed with drugs for 40 days, the action of SOD and GSH Px of brain tissues were determined. Bissecting and fecthing the brain of the adult and aging rats and preparing brain chondriosomes and microsomes, and MDA content of brain chondriosomes and microsomes by the method of Fe 2+ cysteine inducing oxidation reduction reaction were determined. Results: Ginseng stem leaf saponin can increase learning and memory ability of aging mice, decrease the content of lipid peroxide and inhibit the action of MAO B of brain tissues; increase the action of SOD and GSH Px of brain tissues; and also inhibit the content of LPO induced oxidation reduction reaction by Fe 2+ cysteine in brain chondriosomes and microsomes of the adult and aging rats. Conclusion:Ginseng stem leaf saponin can protect brain function and postpone brain aging by decreasing free radical's damage and increasing action of GSH Px and SOD of brain tissue.
8.An analysis of the clinical features of 424 cases of hepatitis E
Zhenping FAN ; Wenjin ZHANG ; Feng GAO
Medical Journal of Chinese People's Liberation Army 2001;0(08):-
171.1?mol/L) was 25.0%, 37.7% and 46.3%, the percentage of the fulminating hepatitis was 3.0%, 5.9% and 14.0%, and the mean hospital stay days for the convalescent patients were 31.59?18.97, 31.13?13.70 and 37.51?18.33 days, respectively. All these variables were significantly higher in aged group compared with youth and middle-aged groups. The levels of alanine transaminase (ALT) lowered significantly with the advance of age, and they were 1 711.7?1 063.4, 1 423.0?913.2 and 1 162.7?792.5U/L, respectively. The same was true in serum albumin and choline esterase (ChE). On the other hand, the percentage of acute hepatitis without jaundice decreased to 17.9%, 6.4%and 2.5% from the youth to the aged. However, TBIL on admission (131.41?106.97 vs 169.60?136.11 vs 164.36?106.22?mol/L) and the elevation of the peak of TBIL (135.01?109.15 vs 186.08?150.64 vs 209.63?143.74?mol/L) of the middle-aged and the aged were significantly higher than those of the youth. There was no significant difference in the symptoms, the ratio of direct to total bilirubin, mortality to complications among them. Furthermore, the serum IgM and IgG antibodies against HEV were negative in about 8.1% of the patients with HEV. Conclusion With a higher incidence and more severe hepatic damage, the middle-aged and the aged patients need a longer duration for recovery than the young patients.
9.Drug-resistance of bacteria and application of antibacterial agents in spontaneous bacterial peritonitis
Wenjin ZHANG ; Zhenman WEI ; Xiaofeng WANG
Medical Journal of Chinese People's Liberation Army 2001;0(09):-
Objective To investigate the distribution of the bacteria in ascites of spontaneous bacterial peritonitis (SBP), and to find out the resistance of the bacteria to antibacterial agents. Methods 44 inpatients from 1999 to 2003 with SBP and positive bacterial culture in ascites were selected, and the bacterial distribution, resistance to drugs and the application of antibacterial agents were analyzed. Results 88.6% (39/44) of the 44 isolations were gram-negative bacillus, among them 28 isolations were E. coli. The results of drug sensitive test revealed that 90% of E. coli, Aeromonas sobria, Klebsiella pneumoniae were sensitive to the third generation of cephalosporin. The resistant rate of E. coli to cefotriaxone and cefotaxime were 11.54% and 12.50%, respectively. Conclusion The third generation of cephalosporin was the experiential selection in treating SBP before getting the report of drug sensitive test. Anti-bacterial agents combined with ?-lactamase inhibitors may be used in treating some severe cases.
10.Effects of C3aR agonists on the phenotype of mouse primary renal tubular epithelial cell
Zhicheng ZHANG ; Wenjin ZHAO ; Jingmin ZHENG
Journal of Medical Postgraduates 2017;30(1):14-20
Objective The key points in the epithelial-mesenchymal transition ( EMT) procedure include the downregula-tion of epithelial protein (E cadherin) and the upregulation of cell activity and cell matrix generation .The aim of this study was to es-tablish a method for primary culture and identification of mouse renal tubular epithelial cells and to explore whether the activation of C3aR can induce epithelial-to-mesenchymal transition in mouse primary renal epithelial cells . Methods Murine renal tubular seg-ments were used for primary cell culture .Immunocytochemistry and immunofluorescence staining were used to identify the renal tubular epithelial cells.The experiment groups included control group , five different concentrations of C3aR agonist groups (0.1, 1, 100, 500, and 2000 ng/mL), and three different time-point groups.The mRNA levels of E-cadherin,α-smooth muscle actin (SMA) and colla-gen I in renal tubular epithelial cells were detected by Real-time PCR; the protein of E-cadherin, α-SMA were detected by Western blot.The cytoskeleton of epithelial cells was observed by phalloidin staining . Results Compared with the control group , the protein expression of E-cadherin deceased (0.950±0.901 vs 0.650±0.221) and the expression of α-SMA (1.380±0.062 vs 1.600±0.103) and collagen I increased in C3aR agonist group (500 ng/mL, after 48 hours) (P<0.05).In addition, the association between these changes and C3aR agonists was presented in a dose-and time-dependent man-ner, respectively.The cytoskeleton staining showed that treatment of renal tubular epithelial cells with C 3aR agonists induced the formation of actin stress fibers in a time-dependent manner . Conclusion The method for primary culture and identification of mouse renal tubular epithelial cells were successfully established .The activation of C3aR could induce epithelial-to-mesenchymal transition in mouse primary renal epithelial cells , which plays an essential role in the de-velopment of renal fibrosis .Moreover , this study indicated that C 3aR may become a new therapeutic target in kidney diseases .