1.Application of ELISA in Detecting Heparin-Induced Thrombocytopenia Antibodies
Hongmei WANG ; Jingjing TIAN ; Shengbao DUAN ; Shaohua DING ; Qinglin MENG ; Yezhou CHEN ; Qiang CHEN ; Yong LI
Journal of Modern Laboratory Medicine 2014;(6):65-68
Objective To develop an ELISA method for determination of heparin-induced thrombocytopenia (HIT)antibody. Methods The compound formed between human platelet factor 4 (PF4)and heparin was used as the coating antigen,incu-bating the patients plasma with the coating antigen in the well,after washing,the second antibody labeled HRP was added in the well to incubate and washing again,the chromogenic substrates was added in the well to incubate,when the stop reaction was finished,the absorbance A450/A630 was detected,and the test results were judged according to standard,this method was compared with IBL method and was optimized and evaluated the performance.Results An indirect ELISA method was de-velop with the purified human PF4,the optimal dilution of sample and second antibody were 1∶100 and 1∶1 500 which de-tected by the orthogonal test,the intra-and inter-assay average coefficients of variation were 7.66% and 7.76%(<10%) respectively that detected by repeated measurement the three positive standard plasma.Through measureing the 100 healthy human plasm with no history of using heparin,the positive and negative predictive reference values were 0.304 and 0.456. IBL and this method detected 100 hemodialysis patients samples at the same time,and the result of statistical analysis was that,the sensitivity,speciality and accuracy of this method were 90%,97.78% and 90%,respectively.The negative and posi-tive predictive value were 81.8% and 98.88% respectively,and the difference was statistically significant [K=0.84(0.81~1)and Pexac=0.012<0.05].The difference was statistically significant,consistency was optimal,95% confidence interval was 92.59%~92.59%.Conclusion Comparing with the IBL,the method reported by this article had the similar perform-ance and good consistency,and it could satisfy the clinical detection and diagnosis of HIT patients.
2.Study of the etoricoxib combined with Bitong decoction formula on the clinical effect and serum level in patients with knee osteoarthritis
Yezhou LI ; Chengjian TANG ; Pei CHEN ; Qing CHEN
Chinese Journal of Pharmacoepidemiology 2024;33(11):1201-1208
Objective To explore the effect of etoricoxib combined with Bitong decoction formula on the clinical efficacy and serum level of patients with knee osteoarthritis.Methods Retrospective analysis was used to collect patients with knee osteoarthritis admitted to our hospital from August 2022 to August 2023.According to the medication method,they were divided into the combined group and the control group.The control group was given etoricoxib tablets,and the combination group was given a combination of Bitong decoction formula on the basis of the control group.Both groups were treated for one month.The efficacy after treatment,Traditional Chinese Medicine(TCM)syndrome score,the knee joint function[osteoarthritis index(WOMAC),Lequesne score],inflammatory indicators[interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),matrix metalloproteinase-3(MMP-3),tissue inhibitor of metalloproteinase-1(TIMP-1)],oxidative stress indicators[total antioxidant capacity(TAOC),lipid peroxidation(LPO),nitric oxide(NO)]before and after treatment and safety of the two groups were compared.Results A total of 162 patients were included,with 81 in each group.The overall efficacy and total effective rate of the combined group were higher than those of the control group(P<0.05).After treatment,TCM syndrome score,WOMAC,Lequesne score,IL-1 β,TNF-α,MMP-3,TIMP-1,LPO and NO levels in two groups were decreased compared with before treatment,while TAOC increased compared with before treatment(P<0.05),and all the above indexes in the combination group were better than those in the control group(P<0.05).The safety of the two groups was comparable(P>0.05).Conclusion The efficacy of etoricoxib combined with Bitong decoction formula in knee osteoarthritis is better than that of single western medicine,which can alleviate symptoms and improve knee joint function,as well as reduce inflammation and oxidative stress,and has high safety.
3.Typical case analysis of COVID-19 cluster epidemic in Shaanxi, 2020
Sa CHEN ; Yi ZHANG ; Chao LI ; Shaoqi NING ; Xinxin LI ; Ni ZHU ; Yunpeng NIAN ; Lei CAO ; Guojing YANG ; Weihua WANG ; Yezhou LIU ; Liang WANG ; Fangliang LEI ; Feng LIU ; Mingwang SHEN
Chinese Journal of Epidemiology 2020;41(8):1204-1209
Objective:By analyzed the transmission patterns of 4 out of the 51 COVID-19 cluster cases in Shaanxi province to provide evidences for the COVID-19 control and prevention.Methods:The epidemiological data of RT-PCR test-confirmed COVID-19 cases were collected. Transmission chain was drawn and the transmission process was analyzed.Results:Cluster case 1 contained 13 cases and was caused by a family of 5 who traveled by car to Wuhan and returned to Shaanxi. Cluster case 2 had 5cases and caused by initial patient who participated family get-together right after back from Wuhan while under incubation period. Cluster case 3 contained 10 cases and could be defined as nosocomial infection. Cluster case 4 contained 4 cases and occurred in work place.Conclusion:Higher contact frequency and smaller places were more likely to cause a small-scale COVID-19 cluster outbreak, with potential longer incubation period. COVID-19 control strategies should turn the attention to infection prevention and control in crowded places, management of enterprise resumption and prevention of nosocomial infection.
4.Epidemiological characteristics of COVID-19 in Shaanxi province
Ni ZHU ; Chao LI ; Shaoqi NING ; Sa CHEN ; Lei CAO ; Guojing YANG ; Xinxin LI ; Yunpeng NIAN ; Weihua WANG ; Yezhou LIU ; Liang WANG ; Fangliang LEI ; Yi ZHANG ; Guihua ZHUANG
Chinese Journal of Epidemiology 2020;41(9):1411-1414
Objective:To understand the incidence trend and epidemiological characteristics of COVID-19 in Shaanxi province.Methods:The incidence data of COVID-19 reported in Shaanxi as of 22 February, 2020 were collected for an epidemiological descriptive analysis.Results:A total of 245 confirmed cases of COVID-19 were reported in Shaanxi. Most cases were mild (87.76%). As time passed, the areas where confirmed cases were reported continued to increase. The case number in Xi’an was highest, accounting for nearly half of the total reported cases in the province. The epidemic pattern in Shaanxi had gradually shifted from imported case pattern to local case pattern, and the transmission of local cases was mainly based on family cluster transmission. The confirmed cases from different sources had caused the secondary transmission in Shaanxi. After February 7, the number of reported cases began to fluctuate and decrease stably, indicating a decrease-to-zero period.Conclusions:At present, the overall epidemic of COVID-19 in Shaanxi has gradually been mitigated. However, considering the approaching of return to work and study and the increasing of imported cases from other countries, the prevention and control of COVIS-19 in Shaanxi will face new challenges.
5.Establishment of CD36 negative platelet donor bank in Zhongshan area
Yonglun WU ; Ainong SUN ; Fei PU ; Qiao LI ; Yuru FANG ; Qianying CHEN ; Yanting LIAO ; Hongmei WANG ; Yezhou CHEN ; Shengbao DUAN
Chinese Journal of Blood Transfusion 2022;35(5):558-561
【Objective】 To investigate the frequency of CD36 deletion and gene mutation in voluntary blood donors of Zhongshan city, and to explore the possibility of establishing local CD36 negative platelet donor bank. 【Methods】 Platelet CD36 antigen was detected by ELISA in 1 654 voluntary blood donors.Some of the negative samples were confirmed by flow cytometry, and genotyping was also performed. 【Results】 Platelet CD36 antigen was negative in 27 cases, accounting for 1.6% (27/1654), among which 1.6% (18/1149) were males and 1.8% (9/505) were females.No significant difference was noticed between males and females in CD36 antigen deletion cases (P>0.05). Fifteen CD36 negative samples were randomly selected, genotyped and sequenced, with type I deletion in 1 case[ 6.7% (1/15)], type Ⅱ deletion in 14 cases[ 93.3% (14/15)], and gene mutation in exon 3-14 detected in 8 cases. 【Conclusion】 The frequency of platelet CD36 antigen deletion in Zhongshan is comparable to that in other southern regions of China.The establishment of CD36 negative platelet donor bank is conductive to improve the effectiveness of platelet transfusion.
6.Platelet antibody of blood donors in Suzhou
Li DONG ; Ming FANG ; Yujue WANG ; Honghong HE ; Hongmei WANG ; Yezhou CHEN ; Feiran WU ; Shengbao DUAN ; Longhai TANG
Chinese Journal of Blood Transfusion 2022;35(8):795-799
【Objective】 To study the incidence and specificity of platelet antibody in blood donors in Suzhou, analyze the distribution characteristics of platelet antibody in blood donors in this area, and explore the significance of platelet antibody detection in blood donors to reduce the adverse reactions toplatelet transfusion in clinical. 【Methods】 Platelet antibody detection was performed in 2178 blood donors in this area by solid-phase immunosorbent assay. The antibody specificity of the positive samples was analyzed by commercial kit, and the anti-CD36 antibody positive samples were further identified by flow cytometry and gene sequencing. 【Results】 Twelve positive samples were detected by platelet antibody screening, with a positive rate of 0.55%(12/2 178), including 5 males (0.33%, 5/2 178)and 7 females(1.06%, 7/2 178). Among the positive samples, anti-HLA-Ⅰ antibody was identified in 2 cases, anti-CD36 antibody in 1 case, and the antibody specificity was not identified in the other 9 cases. In one case, the positive rate of anti-HLA-Ⅰ antibody PRA was 31.31%(31/ 99), which was mainly specific to anti-B15, anti-B35 and anti-B40. The positive rate of anti-HLA-Ⅰ antibody PRA in the other case was 45.45%(45/ 99), which was mainly specific to anti-A2, anti-A11, anti-A24, anti-A29, anti-A33, anti-A66, anti-B15 and anti-B35. The blood donor with anti-CD36 antibody was type I CD36 deficiency, and 329_330delAC mutation occurred in exon 5. 【Conclusion】 Through antibody screening and specificity identification, the positive rate of platelet antibody in females was significantly higher than that in males(P<0.05). In addition to the common anti-HLA-I antibodies, anti-CD36 antibody was also detected in type I CD36 deficient blood donor. Therefore, the detection of platelet antibodies in blood donors is of certain clinical significance to reduce the adverse reactions to blood transfusion caused by antibodies in platelet products.
7.Phenotype screening and gene mutation analysis of platelet CD36 antigen: blood donors in Suzhou area
Honghong HE ; Jing CHEN ; Yezhou CHEN ; Yujue WANG ; Longhai TANG ; Hongmei WANG ; Shaohua DING ; Shuangshi WEI ; Jingjing TIAN ; Shengbao DUAN
Chinese Journal of Blood Transfusion 2021;34(6):585-589
【Objective】 To investigate the expression of CD36 antigen in Suzhou area, analyze the type of antigen deficiency and gene mutation, so as to provide references for the establishment of CD36 negative donor registry in Suzhou. 【Methods】 Anticoagulant whole blood samples (805 cases) were randomly collected from healthy blood donors in Suzhou Blood Center. The expression of CD36 antigen on platelet and monocyte was analyzed by flow cytometry to determine the type of CD36 deficiency. The gene mutation type of platelet CD36 antigen-deficient was performed by genomic DNA sequencing. 【Results】 The CD36 deficiency frequency on platelet was 2.48% (20/805), among which TypeⅠ(lacking CD36 expression both on platelet and monocyte) and TypeⅡ(lacking CD36 expression on platelet only) CD36 deficiency accounted for 10% (2/20) and 90% (18/20), respectively. CD36 gene mutations were found in 10 samples, including 3 cases of 329_330 delAC, 1 case of 1228_1239 delATTGTGCCTATT and 2 cases of 1163 A>T; 1 case of 329_330 delAC+ 1172_1183 delTATTGGTCAAGC and 287 G>C+ 329_330 delAC heterozygous mutation. In addition, 1 case of 745 A>G and 1 case of 806 C>T mutations were novel, and not yet reported. 【Conclusion】 Results showed that the frequency of CD36 antigen deficiency in Suzhou were similar to that reported in southern China, but the mutation sites were slightly different. The establishment of CD36 negative platelet registry could provide negative platelets for patients with transfusion reactions caused by anti-CD36 antibody and improve the effect of clinical platelet transfusion.
8.Effect of hydrogen sulfide on TGF⁃ β1mediated epithelial⁃mesench ymal transformation in bleomycin⁃induced pulmonary fibrosis rats
Rongfang Tu ; Saili Zeng ; Zhenhua He ; Xiaowu Tan ; Zhe Chen ; Yezhou Xia ; Xuehua Li
Acta Universitatis Medicinalis Anhui 2022;57(1):77-83
Objective :
To observe the effect of hydrogen sulfide( H2 S) on the expression of transforming growth factor⁃β1(TGF⁃ β1) , E ⁃cadherin (E⁃CAD) , Vimentin (VIM) , alpha⁃smooth muscle actin ( α ⁃SMA) during epithelial⁃mesenchymal transformation(EMT) , and to explore the anti⁃fibrosis mechanism of it.
Methods :
Sixty rats ( male SD) were randomly divided into control group, bleomycin group, NaHS + bleomycin group and prednisolone + bleomycin group , 15 rats per group. 5 rats of each group were sacrificed at random in 7th , 14th and 28th day. The degree of alveolitis and pulmonary fibrosis was observed . The expression of protein and mRNA of TGF⁃ β1 , E ⁃cad , VIM , α ⁃SMA were determined by Immunohistochemi stry and RT⁃PCR.
Results :
① HE and Masson staining showed that the lung tissue of fibrosis had the lowest degree in control group. and the most severe in bleomycin group. The lung tissue of NaHS + bleomycin group and prednisolone + bleomycin group also had alveolitis and fibrosis changes , but the degree Were significantly less than bleomycin group. ② The mRNA and protein expression levels of TGF⁃ β1 , VIM and α ⁃SMA in bleomycin group, NaHS + bleomycin group and prednisolone + bleomycin group were all higher than that in control group at 7th , 14th and 28th day ( P < 0. 05 ) , while the expression levels of them in NaHS + bleomycin group and prednisolone + bleomycin group were all lower than that in bleomycin groupat each time (P < 0. 05) , which was significant at 28th day in NaHS + bleomycin. ③ The mRNA and protein expression levels of E ⁃Cad in bleomycin group, NaHS + bleomycin group and prednisolone + bleomycin group were all lower than that in control group at 7th , 14th and 28th day(P < 0. 05) , but the expression levels of E ⁃Cad in⁃NaHS + bleomycin group and prednisolone + bleomycin group were higher than that in bleomycin group at each time(P < 0. 05) , which was significant at 28th day in NaHS + bleomycin.
Conclusion
H2 S can reduce the degree of pulmonary fibrosis in rats , its mechanism may be related to the down⁃regulation of TGF⁃ β1and the inhibition of the EMT , which can enhance the expression of E ⁃cad and reduce the expression of TGF⁃ β1 , VIM and α ⁃SMA.