1.Establishment and functional analysis of CHO cells with stable expression of hGHR gene and its mutants
Lihua WU ; Xiaoli CHEN ; Wenqin TANG ; Ting ZHANG ; Zhiyong LIAO
Basic & Clinical Medicine 2010;30(1):13-18
Objective To eonstruct three eukaryotic expression vectors containing wilde-type hGHR gene and its mutants(hGHR-E42K and hGHR-H56R) related to congenital growth hormone insensitivity, then check their expression in CHO cells. Methods With the available PUC-hGHR vector, two mutate hGHR genes (hGHR-E42K and hGHR-H56R) were obtained through mutagenesis. Then three recombinants were cloned into eukaryotic expression vectors pcDNA3.1/zeo(+) with restriction enzymatic reactions.Then with Lipofectamine2000, we trans-fected expression vectors to CHO cells and screened the stably expressed CHO cells by Zeocin. RT-PCR and/or Western blotting were used to examine hGHR and STAT5-P. Results After sequencing, two mutations were introduced to hGHR, three eukaryotic expression vectors were identified. The transfected CHO cells expressed vd-hGHR or its mutants. Compared with hGH-wt, two mutate cells (E42K and H56R) had decreased phosphorylated STAT5 levels. Conclusion Three CHO cells which stably expresses wide type hGHR and its mutants were successfully established, E42K and H56R partly interfered the phosphorylation of STAT5.
2.Establishment and functional analysis of CHO cells with stable expression of hGHR gene and its mutants
Lihua WU ; Xiaoli CHEN ; Wenqin TANG ; Ting ZHANG ; Zhiyong LIAO
Basic & Clinical Medicine 2006;0(01):-
Objective To construct three eukaryotic expression vectors containing wilde-type hGHR gene and its mutants(hGHR-E42K and hGHR-H56R) related to congenital growth hormone insensitivity,then check their expression in CHO cells. Methods With the available PUC-hGHR vector,two mutate hGHR genes (hGHR-E42K and hGHR-H56R) were obtained through mutagenesis. Then three recombinants were cloned into eukaryotic expression vectors pcDNA3.1/zeo(+) with restriction enzymatic reactions. Then with Lipofectamine2000,we transfected expression vectors to CHO cells and screened the stably expressed CHO cells by Zeocin. RT-PCR and/or Western blotting were used to examine hGHR and STAT5-P.Results After sequencing,two mutations were introduced to hGHR,three eukaryotic expression vectors were identified. The transfected CHO cells expressed wt-hGHR or its mutants. Compared with hGH-wt,two mutate cells (E42K and H56R) had decreased phosphorylated STAT5levels. Conclusion Three CHO cells which stably expresses wide type hGHR and its mutants were successfully established,E42K and H56R partly interfered the phosphorylation of STAT5.
3.Risk perception, emergency preparedness and emergency literacy among medical students under COVID-19 epidemic
YU Xingli, LI Ailing, TANG Jian, XIE Hong, ZHU Jing, XIONG Wenqin, CHEN Yanhua
Chinese Journal of School Health 2020;41(11):1646-1649
Objective:
To investigate risk perception, emergency preparedness and emergency literacy among medical students under COVID-19 epidemic.
Methods:
An online survey was conducted by using the self designed COVID-19 questionnaire on risk perception, emergency preparedness and emergency literacy from March 27 to March 30, 2020. The convenience sampling method was used to investigate 4 864 medical students from 2 colleges and technical secondary schools in Sichuan Province.
Results:
The scores of risk perception, emergency preparedness and emergency literacy associated with COVID-19 were(35.18±4.33)(28.30±4.16) and(25.23±2.97) respectively. Risk perception of medical students was positively correlated with emergency preparedness and emergency literacy(r=0.35, 0.40), emergency preparedness was positively correlated with emergency literacy(r=0.51)(P<0.05).
Conclusion
Considering interrelationships among risk perception, emergency preparedness and emergency literacy associated with COVID-19, medical students should be given targeted prevention and control training.
4.Specialty construction and accomplishment of rehabilitation medicine in Fenglin Community Health Service Center of Shanghai Xuhui District
Haijiao LIU ; Yangyang WEI ; Peng ZHOU ; Bin XUE ; Lan YANG ; Yinghua WU ; Jie QIN ; Jie DU ; Zhengyong TANG ; Dan QIN ; Wenqin GU
Chinese Journal of General Practitioners 2021;20(3):362-365
Rehabilitation medicine is one of the most important specialties in community health institutions. This article introduces the 12 year′s development of rehabilitation medicine in Fenglin Community Health Service Center, focusing on the talent allocation, service capabilities, resource expansion, basic facilities, personnel recruiting, department operating, service scope, and its achievements and influence, to provide reference for planning and construction of featured specialty in community health service centers.
5. The study of expression and prognostic value of CD123 in acute myeloid leukemia bone marrow blasts
Wenqin YUE ; Gusheng TANG ; Min LIU ; Hui CHENG ; Jing DING ; Tao WANG ; Jianmin WANG ; Xiaoxia HU ; Weiping ZHANG ; Li CHEN ; Jianmin YANG
Chinese Journal of Hematology 2017;38(10):876-882
Objective:
To study the expression of CD123 in bone marrow (BM) blasts of acute myeloid leukemia (AML) patients to explore the relationship between CD123 expression and therapeutic response and prognosis.
Methods:
This study retrospectively analyzed expression and distribution of CD123 in BM blasts in 137 cases of newly diagnosed AML (excluded M3) , CD123 detected by flow cytometry≥20% was defined as positive, including 84 CD123+ AML and 53 CD123- AML, efficacy and prognosis were compared between the two groups.
Results:
① Among 137 patients, 84 were in group CD123+ (61.3%) , and 53 in group CD123- (38.7%) . All 137 patients were classified into risk groups based on cytogenetic and molecular biology abnormalities. No significant differences were seen between the three risk groups with regard to their CD123 levels (
6.The nucleus origin of acoustically evoked short latency negative response in guinea pigs.
Shaohua ZHANG ; Wenqin HUANG ; Chenxi LIN ; Jin CHEN ; Dongxiao NONG ; Anzhou TANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2014;49(10):848-853
OBJECTIVEThis study established a model of acoustically evoked short latency negative response (ASNR) in guinea pigs. Stereotaxic coordinate guided electrolytic lesion was applied to animal brainstem nuclei, the vestibular nucleus and the cochlear nucleus, to define the neural origin of ASNR.
METHODSTwenty four guinea pigs with normal hearing were randomly divided into the control group (8 subjects, 16 ears) and the deafened group (16 subjects, 32 ears). Each animal experienced the auditory brainstem response (ABR) test. According to the presence of ASNR, the deafened group was further divided into ASNR group and non-ASNR group. Electrolytic lesion was conducted to the vestibular nucleus and cochlear nucleus respectively, followed by ABR test. The lesion structures were verified by brainstem slice and microscope.
RESULTSIn deafened group, the ASNR was elicited in 10 ears (31.3%). The ASNR was eliminated due to the electrolytic destruction to the vestibular nucleus, but it remained unchanged after the same procedure to the cochlear nucleus.
CONCLUSIONIt is clear that the ASNR is originated from the vestibular nucleus, but not the cochlear nucleus.
Acoustics ; Animals ; Cochlear Nucleus ; Evoked Potentials, Auditory, Brain Stem ; Guinea Pigs ; Reaction Time ; Saccule and Utricle ; Vestibular Nuclei
7.Investigation on the Mechanism of Salvia miltiorrhiza in the Treatment of Postoperative Abdominal Adhesion Based on Network Pharmacology and Molecular Docking
Wenqin LIU ; Fuling WU ; Long WANG ; Qin YANG ; Jiangjie WU ; Lianbing HOU ; Lan TANG ; Chuqi HOU
China Pharmacy 2021;32(24):2987-2993
OBJECTIVE:To investigate the potential mechanism of Salvia miltiorrhiza in the treatment of postoperative abdominal adhesion (PAA). METHODS :Active components and target genes of S. miltiorrhiza were retrieved from TCMSP database,SwissADME database ,Perl database ,UniProt database and other databases. GeneCards ,OMIM and PubMed database were used to retrieve target genes related to PAA. Venn diagram was drawn by using mapping tool of bioinformatic online database so as to screen the intersecting targets of active component-PAA. STRING platform was adopted to establish target network related to active component-PAA and protein-protein interaction (PPI)network of intersecting targets ,etc.,and to screen hub genes. Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genom es(KEGG)pathway enrichment were carried out by using R 3.6.1 software. Using the protein encoded by hub gene as receptor and tanshinone Ⅱ A as ligand ,the molecular docking was carried out with AutoDock 1.5.6 tool. RESULTS :A total of 38 active components of S. miltiorrhiza with high gastrointestinal absorption and their corresponding 72 targets,755 PAA-related target genes were identified. Results of Venn diagram showed that there were 33 intersecting targets of active components of chuqi90@163.com S. miltiorrhiza with PAA. Tanshinone ⅡA,dihydrotanshinolac- tone and other components may be important nodes of the target network related to active component-PAA. FOS,APP,ACHE, CASP3 and PTGS2 may be the hub genes in PPI network of intersecting targets. Results of GO enrichment showed that the intersecting targets were mainly concentrated in adrenergic receptor activity ,catecholamine binding ,G protein-coupled amine receptor activity and so on ;KEGG pathway enrichment analysis showed that the intersecting targets were mainly enriched in neuroactive ligand-receptor interaction ,cGMP-PKG signaling pathway ,endocrine resistance ,EGFR-tyrosine kinase inhibitor resistance and calcium signaling pathway.Molecular docking analysis showed that tanshinone ⅡA could form hydrogen bonds with many amino acid residues such as VAL- 580 of proto oncogenes c-Fos ,amyloid precursor protein ,acetylcholinesterase,caspase 3 and prostaglandin G/H synthase 2. CONCLUSIONS :The active components of S. miltiorrhiza play a role in the treatment of PAA by directly or indirectly acting on neuroactive ligand-receptor interaction ,cGMP-PKG signaling pathway ,endocrine resistance , EGFR-tyrosine kinase inhibitor resistance resistance and calcium signaling pathway.