1.Hb-Enzyme Release Assay for Macrophage-Mediated Aptibody-Dependent Cellular Cytotoxicity
Chinese Journal of Immunology 1986;0(04):-
An improved assay for quantitation of murine macrophage-mediated antibody-dependent cellular cytotoxicity (MMADCC) by hemoglobin-enzyme release assay (Hb-ERA) has been developed The method is based on the coloration measurement by spectrophotometer,because hemoglobin has peroxidase activity capable of catalyzing the oxidation of o-phenylenediamine and producing color reaction. This method was applied to demonstrate variation of MMADCC activity with varying effector:target ratio,incubation time,antibody concentration and macrophage in different stage of activation. The method provides advantages of(l)elimination of the need for expensive and hazardous radioactive materials,(2)relative ease and rapidity, (3)sensitiyity and reproducibility.
2.Interventional treatment of the complications occurring after renal transplantation: a clinical study
Chen WANG ; Sheng LIU ; Qingquan ZU ; Chungao ZHOU ; Xinlong LIU ; Bing WANG ; Chun ZHOU ; Haibin SHI
Journal of Interventional Radiology 2017;26(7):597-600
Objective To evaluate the safety and effectiveness of multi-mode interventional therapy for complications occurring after renal transplantation.Methods The clinical data of 26 patients with complications occurring after renal transplantation were retrospectively analyzed.Vascular and non-vascular complications were treated with different interventional techniques,and the curative effects were analyzed.Results Vascular complications were observed in 19 patients and non-vascular complications were seen in 7patients.The technical success rate of interventional therapy was 100%.One week after the treatment,the total creatinine level (SCr) was significantly improved,which decreased from preoperative (372.7±295.5)μmol/L to postoperative (184.3±138.4) μmol/L (P<0.001).No severe complications occurred.Further analysis indicated that no statistically significant differences in patient's general condition,kidney donor source,anastomosis method existed between vascular intervention group and non-vascular intervention (P>0.05).However,the onset time of vascular complications was markedly earlier than that of non-vascular complications (1.8± 1.4 months vs.118.3 ±54.4 months),the difference was statistically significant (P<0.001).During the follow-up period lasting for 4-55 months (mean of 18.6 months),3 patients developed recurrence of complications;interventional therapy had to be carried out in 2 patients and their creatinine level returned to normal after treatment,and transplanted renal artery embolization had to be performed in the other patient as whose pseudoaneurysm became enlarged.Conclusion For the treatment of complications occurring after renal transplantation,interventional therapy is less-invasive,rapidly-effective and safe,this technique can timely and effectively improve the renal function and save the transplanted kidney.
3.Study of mitochondrial DNA A1555G mutation among nonsyndromic hearing impairment in Chinese population
Qi-Shui OU ; Zu-Jian CHENG ; Jing CHEN ; Bin YANG ; Ling JIANG ; Sheng-Nan YE ;
Chinese Journal of Laboratory Medicine 2001;0(03):-
Objective To study the prevalence of the mtDNA A1555G gene mutation in Chinese population with nonsyndromic hearing impairment.Methods PCR-RFLP,directional sequencing of PCR products were applied in 325 patients with nonsyndromic hearing impairment and 50 normal controls.Results The mutation rate of the mtDNA A1555G was 14.5% (47/325),28 of 47 cases were homozygosis,19 of 47 cases were heterozygosis.The same mutation was not detected in the control subjects.Conclusion The mutation rate of the mtDNA A1555G is relatively high in the Chinese NSHI patients,the mutation type includes both heterozygosis and homozygosis.
4.Application and evaluation of droplet digital polymerase chain reaction in detection of severe acute respiratory syndrome corona virus-2
CHEN Shuang ; WANG Ming-yue ; ZU Zhen ; TANG Yun ; YE Sheng ; LING Hua ; TAN Zhang-ping
China Tropical Medicine 2023;23(5):495-
Abstract: Objective To analyze the nucleic acid detection results of severe acute respiratory syndrome corona virus-2 (SARS-CoV-2) by droplet digital PCR (ddPCR) and compare with the detection results of real-time fluorescence quantitative RT-PCR (qRT-PCR), so as to evaluate the advantages and disadvantages of detection, and to provide data support for optimizing the nucleic acid detection scheme of SARS-CoV-2. Methods According to the SARS-CoV-2 specific primer probe published by the China Center for Disease Control and Prevention, a ddPCR detection method for SARS-CoV-2 was designed. One sample was selected for sensitivity test after gradient dilution; six respiratory virus nucleic acid positive samples including seasonal H3N2 influenza virus and SARS-CoV-2 positive samples were selected for specificity test; five SARS-CoV-2 positive samples were selected for repeatability test; in addition, 30 positive and 20 negative SARS-CoV-2 samples were selected for multiple clinical samples testing, and the results were analyzed and compared with those of qRT-PCR. Results The ddPCR method can specifically detect SARS-CoV-2, and directly obtain the original copy number of the sample target gene to achieve accurate quantification; the sensitivity test of gradient dilution positive samples showed that qRT-PCR detected target genes in part of the 10-5 dilution of samples, and no target genes were detected in 10-6 dilution, while ddPCR detected all target genes in both 10-5 and 10-6 dilution of samples. The detection limit of ddPCR was two orders of magnitude higher than that of qRT-PCR, and the sensitivity was higher than that of qRT-PCR; in the comparison of the repeatability test results of the two methods, the coefficient of variation of ddPCR was 1.266%-11.814%, lower than 1.729%-26.174% of qRT PCR, and the repeatability was higher than qRT-PCR; among 50 clinical samples, 30 positive samples of confirmed cases of Coronavirus Disease 2019 (COVID-19) were detected by both methods, SARS-CoV-2 was successfully detected by both methods, and 20 negative samples of COVID-19 were detected by both methods, and the results were negative, with a coincidence rate of 100.00% (50/50). Conclusion The ddPCR method can accurately quantify SARS-CoV-2 with strong specificity, and its sensitivity and repeatability are higher than those of qRT-PCR, but it also has certain detection limitations and is more suitable for the detection of low load samples. In the actual detection, the two methods can be reasonably combined to improve the detection accuracy.
5.Construction of a novel Schistosoma japonicum DNA vaccine pBK-Sj14-3-3 and studies on its immunoprotection in mice.
De-fa LI ; Yue-sheng CHEN ; Ying ZU ; Ji-long SHEN
Chinese Journal of Preventive Medicine 2004;38(3):193-195
OBJECTIVETo prepare Sj14-3-3 DNA vaccine and observe its immunoprotection against Schistosoma japonicum in mice.
METHODSThe Sj14-3-3 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and subcloned into eukaryotic expression vector pBK. The recombinant plasmid pBK-Sj14-3-3 was extracted, purified and inoculated into BALB/c mice by intramuscular injection. Mice were attacked by Schistosoma japonicum cercariae and then killed. Adult worm and egg were counted, respectively. Diameter of the egg granulomas in the liver of infected mice was measured.
RESULTSElectrophoresis on 1% agarose gel showed that the product of RT-PCR and the inserted fragment of recombinant plasmid digested with EcoR I and Xho I had the same size, about 765 bp, confirming the latter was the 14-3-3 encoding gene by nucleotide sequencing. Adult worm load declined by 27%, average egg load of per gram (EPG) of the liver tissues by 79%, average egg production per couple of adult worm (EPWP) by 51%, and mean diameter of egg granulomas by 29% in vaccinated mice.
CONCLUSIONThe recombinant plasmid pBK-Sj14-3-3 was successfully constructed, which had some immunoprotection against Schistosoma japonicum in infected mice, indicating its potential to be vaccine candidate molecule of Schistosoma japonicum.
14-3-3 Proteins ; genetics ; immunology ; Animals ; Antibodies, Helminth ; blood ; Antigens, Helminth ; genetics ; immunology ; Cloning, Molecular ; DNA, Helminth ; genetics ; Female ; Helminth Proteins ; genetics ; immunology ; Membrane Proteins ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Parasite Egg Count ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Schistosoma japonicum ; genetics ; immunology ; Schistosomiasis japonica ; immunology ; prevention & control ; Vaccines, DNA ; immunology
6.Prevalence of workplace violence in staff of two hospitals in Guangzhou.
Zu-Hui CHEN ; Sheng-Yong WANG ; Chun-Xia JING
Chinese Journal of Preventive Medicine 2003;37(5):358-360
OBJECTIVETo understand prevalence of workplace violence in hospital and to analyse its relevant causes to lay a basis for maintaining normal working order in hospital.
METHODSA study was conducted to look into workplace violence situation in health care workers in two large hospitals of Guangzhou, Guangdong Province during October 2001 to October 2002. Workplace violence was defined as any events occurred in hospital staff, who suffered psychological or/and physical violence during the past 12 months.
RESULTSTotally, 678 of 1 043 hospital staff (65%) investigated had such experience during the past year, mainly psychological violence. Medical doctors were more vulnerable than nurses, with prevalence of 70.3% and 67.7% for medical doctors and nurses, respectively. Prevalence was the highest in those aged 30 - 39 years with 11 - 20 years of employment. Man staff were more vulnerable to physical violence than women, with prevalence of 11.7% and 5.3%, respectively. No significant difference in psychological or sexual violence between man and woman staff was found. Frequently, nurses and nurse aides were victims of sexual violence. Usually, troublemakers were patients relatives or patients themselves, accounting for 64.2% and 50.0% of the total events, respectively. Main causes for workplace violence in hospital included unreasonable requirement from patients or their relatives which was not met, or not-so-quick recovery as they desired.
CONCLUSIONSWorkplace violence occurred in hospital staff was prevalent in Guangzhou, which should be attached more importance. Comprehensive intervention measures should be adopted focusing on law reinforcement and education, to maintain normal working order in hospital.
Female ; Humans ; Male ; Personnel, Hospital ; Prevalence ; Violence ; prevention & control ; statistics & numerical data ; Workplace
8.Effects of polysorbates on stability of monoclonal antibody drugs
ZHU Sheng-ying ; CAO Jia-wei ; XU Jin ; CHEN Chen-hui ; GUO Qing-cheng ; LI Jun ; ZHANG Da-peng ; QIAN Wei-zhu ; HOU Sheng ; GUO Huai-zu
Chinese Journal of Biologicals 2023;36(2):172-177
Objective To evaluate the effects of various polysorbates(PS)on the stability of different types of monoclonal antibody(mAb)drugs.Methods Three types of monoclonal antibodies mAbA(IgG1 proantibody drug),mAbB(IgG1 mAb)and mAbC(IgG1 mAb with Fc N297A mutation)were used as model proteins,and different kinds or contents of PS were added into the mAb formulations respectively to investigate the influencing factors.The effects of PS on the stability of mAb drugs were evaluated comprehensively by detecting the changes of quality attributes,such as protein aggregates and insoluble particles.Results PS20 and PS80 showed no significant difference in inhibiting the formation of aggregates and charge variants in the three mAbs(P>0.05),while the addition of PS80 in mAbB and PS20 in mAbC significantly inhibited the increase of insoluble particles respectively(P<0.05);The content of PS20 showed a significant effect on the detection indexes of charge variants and insoluble particles in mAbC(P<0.05).Conclusion Different types of mAbs have different sensitivities to various kinds and contents of PS.Therefore,when designing the formulation of mAbs,it is necessary to select appropriate kinds and contents of PS to further improve the stability of mAb drugs.
9.Effect of serum of patients with obstructive jaundice on myogenic differentiation of human pulmona-ry microvascular endothelial cells
Qiwei CHEN ; Yong YANG ; Bing CHEN ; Yang CHEN ; Sheng LI ; Guilan WANG ; Yulong WU ; Chonghui CHEN ; Baoli ZU ; Bin YI ; Kaizhi LU ; Lin LIAO
Chinese Journal of Anesthesiology 2016;36(9):1146-1149
Objective To evaluate the effects of the serum of patients with obstructive jaundice on myogenic differentiation of human pulmonary microvascular endothelial cells (PMVECs). Methods Hu?man PMVECs were isolated and then subcultured. The cultured PMVECs were incubated with the serum of patients with obstructive jaundice or with the serum of healthy volunteers. At 24, 48 and 72 h of incubation (T1?3), the inverted microscope was used to observe the morphology of primary PMVECs. The expression of muscular proteins ( alpha?smooth muscle actin [α?SMA ] , smooth muscle?mysion heavy chain [ SM?MHC] , capolnin) in PMVECs was detected using Western blot analysis. Results The expression of cal?ponin andα?SMA was negative, and a few SM?MHC proteins were expressed when PMVECs were incubated with the serum of healthy volunteers; the expression of calponin, α?SMA and SM?MHC was positive when PMVECs were incubated with the serum of patients with obstructive jaundice. Compared with the serum of healthy volunteers, the expression of SM?MHC was significantly up?regulated when PMVECs were incubated with the serum of patients with obstructive jaundice (P<0.05). The expression of calponin, α?SMA and SM?MHC was significantly up?regulated at T2,3 compared with that at T1 , and at T3 compared with that at T2 when PMVECs were incubated with the serum of patients with obstructive jaundice ( P<0.05) . Conclusion The serum of patients with obstructive jaundice promotes myogenic differentiation of human PMVECs, which is probably one of the mechanisms underlying intrapulmonary microvascular dilatation.
10.Analysis on the epidemiological features and risk factors of hospital workplace violence in Guangzhou.
Zu-hui CHEN ; Sheng-yong WANG ; Ye-cheng LU ; Chun-xia JING
Chinese Journal of Epidemiology 2004;25(1):3-5
OBJECTIVETo study the epidemiological features and relevant risk factors of hospital workplace violence.
METHODSA retrospective study using logistic regression on hospital staff was carried out. Based on WHO's last definition for workplace violence from "new research showing workplace violence threatening health services" issued on May 10th, 2002.
RESULTSAmong 4062 staff members investigated, 2619 had been subjected to workplace violence in the last 12 months preceding the survey, with a rate of 64.48% which was accounted for 49.12% of psychological and 15.36% of physical violence respectively. Male staff were more vulnerable to physical violence against female. Those victims aged 30 - 39 years were at greatest risk. Doctors and nurses were leading victims of workplace violence. Most of workplace violence happened during day shift, in the ward. The main risk factors contributing to workplace violence would include: unreasonable demand from patients or their relatives, late recovery as expected, and expensive cost. Patients' relatives often took the leading role as perpetrators and followed by patients themselves.
CONCLUSIONMagnitude of hospital workplace violence in hospitals in Guangzhou asked for comprehensive measures to improve staff-patient relationship for the prevention and control of violence.
Age Factors ; China ; Female ; Humans ; Logistic Models ; Male ; Personnel, Hospital ; psychology ; statistics & numerical data ; Risk Factors ; Sex Factors ; Surveys and Questionnaires ; Time Factors ; Violence ; statistics & numerical data ; Workplace ; statistics & numerical data