1.Gene Cloning, Construction and Expression of Single-Chain Fv (scFv) Against the Membrane Protein of Schisotosoma japonicum
Xiaocong YU ; Xin JIANG ; Haomin HUANG ; Zhong ZHANG ; Qing LIN ; Xiaohong GUAN ; Hualiang HUANG
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(03):-
Objective To construct single chain antibody specific to membrane protein of Schistosoma japonicum by gonetic engineering technique. Methods The V\-H (heavy-chain variable region) and V\-L (light-chain variable region) genes were amplified by PCR from the genomic DNA of NP11-4 cell line, and sequenced by Sanger's method. The ScFv was constructed in pTHA90 vector using V\-H and V\-L genes, then expressed by IPTG. Results The V\-H and V\-L genes were obtained through PCR. The DNA sequences showed that V\-H and V\-L were new variable region genes of antibody. They were registered by GenBank. A ScFv gene with (Gly4Ser) 3 intralinker in the pTHA90 vector was successfully constructed. The ScFv was expressed as thioredoxin-fused proteins about 36\^2 kDa. Conclusion A specific ScFv against the membrane protein of Schistosoma japonicum was constructed and expressed.
2.Comparison of efficacy of posterolateral and anterolateral locking plate fixation in treatment of extra-articular distal humeral shaft fractures
Wanfu WEI ; Tao YANG ; Fengsong LIN ; Xin LIU ; Zhiming GUO ; Haomin LI ; Desheng ZHAO ; Mingxin LI
Chinese Journal of Trauma 2020;36(2):178-182
Objective:To compare the effect of posterolateral plate and anterolateral locking plate in surgical treatment for extra-articular distal humeral shaft fractures.Methods:A retrospective case-control study was made on 52 patients with extra-articular distal humeral shaft fractures admitted in Tianjin Hospital from January 2014 to October 2016. There were 32 males and 20 females, with the age from 18 to 56 years [(36.9±10.9)years]. According to the AO/OTA classification, there were 15 patients with type A, 32 type B, and 5 type C. Twenty-three patients were treated with osterolateral locking plate (Group A) and 29 with anterolateral locking plate (Group B). Operation time, bone union time, range of motion of the elbow, Mayo elbow performance score and complication rate were evaluated.Results:All patients were followed up for 12-20 months [(13.7±2.2)months]. Operation time was (79.8±9.6)minutes in Group A and (85.0±11.6)minutes in Group B ( P>0.05). Bone union time was (4.1±1.0)months in Group A and (4.1±1.0)months in Group B ( P>0.05). Degrees of elbow extension was 3.9°(0.0°, 5.0°) in Group A, and 4.4°(0.0°, 5.0°) in Group B ( P>0.05). Degrees of elbow flexion was 127.4°(125.0°, 132.50°) in Group A and 128.5°(122.5°, 132.5°) in Group B ( P>0.05). Mayo elbow performance score was 91.0 (90.0, 93.5) points in Group A and 90.2 (90.0, 92.5)points in Group B ( P>0.05). Radial nerve damage was noted after operation, showing no significant difference between two groups [2 patients (9%) in Group A and 3 patients (10%) in Group B] ( P>0.05), and the symptoms were recovered in all patients within 3 months. Conclusions:Both the posterolateral and anterolateral locking plate are effective in surgical treatment for extra-articular distal humeral shaft fractures. However, the posterolateral plate can be placed closer to the distal end of the humerus and the multi-directional locking design is more stable, which has advantages for the patient with fracture line close to the elbow joint.
3.Bioinformatics analysis of the mechanism of lung injury and intervention drugs associated with SARS coronavirus infection
Jundong ZHANG ; Bo YANG ; Lin LIU ; Haoran CHEN ; HaoMin ZHANG ; Bin GUO ; Ximeng CHEN ; Peng ZHI ; Zhuoyang LI ; Geliang LIU ; Xiaohua CHI ; Xuechun LU
Chinese Journal of Experimental and Clinical Virology 2020;34(4):367-373
Objective:To analyze the mechanism and potential intervention drugs of acute lung injury caused by severe acute respiratory syndrome coronavirus (SARS-CoV) infection in order to provide reference for the treatment of COVID-19.Methods:Gene Expression Omnibus (GEO) announcement database was used to screen coronavirus transcriptome data, and R language package was used for differential expression analysis and Kyoto Gene and Genome Encyclopedia (KEGG) and Gene Ontology (GO) enrichment analysis. A protein-protein interaction (PPI) network analysis was carried out by using STRING online analysis website, and the key genes were screened. Then the Epigenomic Precision Medicine Prediction Platform (EpiMed) was used to analyze the association of key genes and predict potential therapeutic drugs.Results:Based on the whole genome expression profile data of SARS-CoV, a total of 3 606 differential genes were screened, including 2 148 up-regulated and 1 458 down-regulated. GO enrichment is mainly related to viral infection, leukocyte migration and adhesion, acute inflammation and collagen secretion. KEGG enrichment is mainly related to signal transduction, acute inflammation, immune response and so on. Ten key genes related to lung injury, such as PTPRC, TIMP1, ICAM1 and IL1B, were screened by protein interaction network analysis. EpiMed platform predicted that pulsatilla chinensis, polygonum cuspidatum, tumor necrosis factor-α inhibitors, famciclovir, fluvastatin and other drugs have potential therapeutic effects.Conclusions:SARS-CoV infection can cause lung injury by activating a series of inflammation-related molecules. Drugs that may be effective in the treatment of coronavirus infections, including pulsatilla chinensis, polygonum cuspidatum, tumor necrosis factor-α inhibitors, famciclovir and fluvastatin.
4.Effect of hypothermic machine perfusion on expression levels of inflammatory cytokines in rat kidney
De LUO ; Jiang LIU ; Pengcheng ZHOU ; Piao WANG ; Xujia LI ; Haomin LIN ; Song SU
Organ Transplantation 2023;14(6):824-830
Objective To evaluate the effect of hypothermic machine perfusion (HMP) on the expression levels of inflammatory cytokines in rat kidney. Methods Thirty male rats were randomly divided into the control (Control group), static cold storage group (SCS group) and HMP group, with 10 rats in each group. The velocity, intrarenal resistance and pH value of perfusion effluent were recorded during HMP. The expression levels of CXC chemokine ligand (CXCL)1, CXCL2, interferon (IFN)-β1, IFN-α4, CC chemokine ligand (CCL)2, CCL20, interleukin (IL)-17α, IL-17C and tumor necrosis factor (TNF)-α messenger RNA (mRNA) in renal tissues were evaluated by reverse transcription polymerase chain reaction (RT-PCR). Pathological changes of the kidney were observed by hematoxylin-eosin (HE) staining. Results During HMP, the velocity and intrarenal resistance remained stable, and the pH value of perfusion effluent was decreased slowly. RT-PCR showed that the relative expression levels of CXCL1, CXCL2, CCL2, CCL20, IL-17α, IL-17C and TNF-α mRNA in the SCS and HMP groups were higher compared with those in the Control group. Compared with the SCS group, the relative expression levels of CXCL1, CXCL2, CCL2, CCL20, IL-17α and TNF-α mRNA were up-regulated in the HMP group (all P<0.05). HE staining revealed that the morphology of renal cells was normal in the Control group, whereas evident epithelial necrosis, cytoplasmic vacuolation, brush border loss and epithelial shedding were observed in the SCS group. Compared with the SCS group, pathological changes in the HMP group were alleviated. Conclusions HMP may activate renal inflammation, and inhibiting the activation of inflammation during HMP is expected to further improve the effect of allograft preservation.
5.Surveillance of antifungal resistance in clinical isolates of Candida spp.in East China Invasive Fungal Infection Group from 2018 to 2022
Dongjiang WANG ; Wenjuan WU ; Jian GUO ; Min ZHANG ; Huiping LIN ; Feifei WAN ; Xiaobo MA ; Yueting LI ; Jia LI ; Huiqiong JIA ; Lingbing ZENG ; Xiuhai LU ; Yan JIN ; Jinfeng CAI ; Wei LI ; Zhimin BAI ; Yongqin WU ; Hui DING ; Zhongxian LIAO ; Gen LI ; Hui ZHANG ; Hongwei MENG ; Changzi DENG ; Feng CHEN ; Na JIANG ; Jie QIN ; Guoping DONG ; Jinghua ZHANG ; Wei XI ; Haomin ZHANG ; Rong TANG ; Li LI ; Suzhen WANG ; Fen PAN ; Jing GAO ; Lu JIANG ; Hua FANG ; Zhilan LI ; Yiqun YUAN ; Guoqing WANG ; Yuanxia WANG ; Liping WANG
Chinese Journal of Infection and Chemotherapy 2024;24(4):402-409
Objective To monitor the antifungal resistance of clinical isolates of Candida spp.in the East China region.Methods MALDI-TOF MS or molecular methods were used to re-identify the strains collected from January 2018 to December 2022.Antifungal susceptibility testing was performed using the broth microdilution method.The susceptibility test results were interpreted according to the breakpoints of 2022 Clinical and Laboratory Standards Institute(CLSI)documents M27 M44s-Ed3 and M57s-Ed4.Results A total of 3 026 strains of Candida were collected,65.33%of which were isolated from sterile body sites,mainly from blood(38.86%)and pleural effusion/ascites(10.21%).The predominant species of Candida were Candida albicans(44.51%),followed by Candida parapsilosis complex(19.46%),Candida tropicalis(13.98%),Candida glabrata(10.34%),and other Candida species(0.79%).Candida albicans showed overall high susceptibility rates to the 10 antifungal drugs tested(the lowest rate being 93.62%).Only 2.97%of the strains showed dose-dependent susceptibility(SDD)to fluconazole.Candida parapsilosis complex had a SDD rate of 2.61%and a resistance rate of 9.42%to fluconazole,and susceptibility rates above 90%to other drugs.Candida glabrata had a SDD rate of 92.01%and a resistance rate of 7.99%to fluconazole,resistance rates of 32.27%and 48.24%to posaconazole and voriconazole non-wild-type strains(NWT),respectively,and susceptibility rates above 90%to other drugs.Candida tropicalis had resistance rates of 29.55%and 26.24%to fluconazole and voriconazole,respectively,resistance rates of 76.60%and 21.99%to posaconazole and echinocandins non-wild-type strains(NWT),and a resistance rate of 2.36%to echinocandins.Conclusions The prevalence and species distribution of Candida spp.in the East China region are consistent with previous domestic and international reports.Candida glabrata exhibits certain degree of resistance to fluconazole,while Candida tropicalis demonstrates higher resistance to triazole drugs.Additionally,echinocandins resistance has emerged in Candida albicans,Candida glabrata,Candida tropicalis,and Candida parapsilosis.