1.Immortalization and screening of cardiac progenitor cells in mice
Gaohui ZHU ; Tongchuan HE ; Feng XIONG ; Min ZHU ; Yuan CHEN
Chinese Journal of Tissue Engineering Research 2014;(23):3715-3722
BACKGROUND:Transplantation of exogenous stem cells for functional cardiac celldeath or apoptosis easily induced complications after transplantation. Therefore, cardiac progenitor cells of heart itself have been ideal seed cells. OBJECTIVE:To establish stable celllines of cardiac progenitor cells from mouse heart, and to provide ideal cellmodels for studying proliferation and differentiation factors affecting cardiac progenitor cells during adult myocardial cells were damaged. METHODS:(1) Myocardiocytes were isolated from embryonic 15.5 days mice. (2) The cultured myocardiocytes were immortalized using retrovirus SSR69. Immortalized monoclonal myocardial cells were obtained using antibiotic selection and infinite dilution. (3) Monoclonal cellline with the property of cardiac progenitor cells was screened out. RESULTS AND CONCLUSION:(1) Myocardiocytes were successful y isolated and cultured. Partial cells showed obvious beating. (2) Myocardiocytes infected with retrovirus SSR69 were cloned and got 76 clones, then were named as CP15-#, (3) Screening the first 20 clones, the reasonable clones with the characteristics of cardiac progenitor cells were obtained according to myocardial cellmarker genes. The results suggested that immortalized cardiac progenitor cells were established mediated by reversible SV40 T antigen.
2.Efficacy on dysphagia induced by bulbar palsy treated with acupuncture at Tiantu (CV 22) with different needling techniques.
Yuan ZHU ; Li-xin FU ; Xue-min SHI
Chinese Acupuncture & Moxibustion 2014;34(11):1089-1092
OBJECTIVETo compare the efficacy between elongated needle therapy and regular needle therapy at Tiantu (CV 22) on the basis of xingnao kaiqiao (activiting brain and regaining consciousness) acupuncture therapy so as to explore the effective therapeutic method in treatment of dysphagia induced by bulbar palsy.
METHODSSeventy one cases of dysphagia induced by bulbar palsy were randomized into two groups. The xingnao kaiqiao acupuncture therapy was applied at Shuigou (GV 26), Neiguan (PC 6), Sanyinjiao (SP 6) and the others in the two groups. In the elongated needle therapy group, on the basis of xingnao kaiqiao acupuncture therapy, the elongated needle was used to puncture Tiantu (CV 22). In the regular needle therapy, the regular acupuncture technique was used at Tiantu (CV 22). In both groups, the treatment was given once a day in a week except Sunday and lasted for 4 weeks totally. Before and after treatment, the swallowing condition and the standardized swallowing assessment (SSA) were observed in the patients and the efficacy was compared between the two groups.
RESULTSThe total effective rate was 97.2% (35/36) in the elongated needle therapy group, which was better than 77.1% (27/35) in the regular needle therapy group (P<0.05). After treatment, SSA score was reduced significantly as compared with that before treatment in the two groups (both P<0.05). SSA score in the elongated needle therapy group was reduced much more apparently as compared with that in the regular needle therapy group after treatment (P<0.05).
CONCLUSIONOn the basis of the xingnao kaiqiao acupuncture therapy, the elongated needle therapy at Tiantu (CV 22) achieves the superior effect on bulbar palsy-induced dysphagia as compared with the regular acu- puncture at Tiantu (CV 22).
Acupuncture Points ; Acupuncture Therapy ; Aged ; Bulbar Palsy, Progressive ; complications ; Deglutition Disorders ; etiology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome
3.Combination of Pdx1 and NeuroD1 results in Nkx6.1 and GLUT2 expressions in L02 cells
Xiaolong TANG ; Min GUO ; Yuan ZHANG ; Kanger ZHU
Chinese Journal of Pathophysiology 1986;0(01):-
AIM: To construct a recombinant plasmid vector containing human pancreatic duodenal homeobox1(Pdx1) and neurogenic differentiation 1(NeuroD1) genes,and to detect its effective expression in eukaryocytes and the ability to induce differentiation of hepatocytes.METHODS: Using human embryo pancreas mRNA as template,Pdx1 and NeuroD1 genes were amplified by RT-PCR and cloned into the two different multiple cloning sites(MCSA and MCSB) of plasmid pIRES.The recombinant plasmid pI/Pdx1/NeuroD1 was transfected into L02 cells.The expression of Pdx1 and NeuroD1 in transfected cells was detected by immunocytochemistry,IFA,RT-PCR and Western blotting,respectively.RESULTS: The length and sequence of cloned segments were correct.Pdx1 and NeuroD1 were expressed in eukaryocytes.Furthermore,the hepatic cells were induced to express glucose transporter 2(GLUT2) and eukaryocyte transcription factor Nkx6.1,which were functionally correlated to ? cells.CONCLUSION: pI/Pdx1/NeuroD1 plasmid is successfully constructed and expressed in human eukaryocytes,with which the cells express the eukaryocyte transcription factor and GLUT2,indicating the transfected cells functionally correlates to ? cells.The results suggest that Pdx1 and NeuroD1 genes can induce the differentiation the cells from hepatic cells to pancreatic endocrine secretion cells.
4.Generation of insulin-producing cells from PDX1 and NKX6.1 gene-modified bone marrow mesenchymal stem cells
Xiaolong TANG ; Yuan ZHANG ; Kanger ZHU ; Min GUO
Chinese Journal of Pathophysiology 2000;0(08):-
AIM:To differentiate bone marrow mesenchymal stem cells(BMSCs) into functional insulin-producing cells to produce sufficient pancreatic islet cells for transplantation.METHODS:Recombinant adenovirus vectors carrying PDX1 and NKX6.1 genes were constructed and the bone marrow mesenchymal stem cells were infected by the recombinant adenovirus combined with several cytokines for differentiation.The expressions of PDX1,NKX6.1 and insulin and C-peptide in the differentiated bone marrow mesenchymal stem cells were detected by RT-PCR and Western blotting.After the differentiated bone marrow mesenchymal stem cells were transplanted into subrenal capsule of diabetic mice,cell morphology of the grafts as well as their secretion of insulin and C-peptide were detected.Besides,regulating capacities of grafts on the blood glucose level of the diabetic mice were also detected.RESULTS:BMSCs induced by recombinant adenovirus(pAdxsi-CMV-PDX1/CMV-NKX6.1) and several cytokines showed positive dithizone staining and the expressions of ?-cell related molecule such as insulin and glucose transporter 2 were detected by RT-PCR,which showed a sustaining and stable expression.The similar results were showed by Western blotting,immunohistochemical staining and indirect immunofluorescence.The insulin secretions in the cells stimulated with glucose at concentrations of 5.5 and 25 mmol/L in the experimental group were(1 240.4?109.3) mU/L and(3 539.8?245.1) mU/L,respectively,and were significantly higher than those in control group.Moreover,transplantation of the cells to STZ mice in treatment group made serum glucose recover to normal level.CONCLUSION:PDX1 and NKX6.1 gene-modified bone marrow mesenchymal stem cells differentiate into insulin-producing cells in vitro.When these cells transplanted into STZ induced diabetic mice,their serum glucose could return to the normal level and they could live well.Thus this is a promising method for diabetes treatment.
5.Epidemiological investigation of diabetic retinopathy in type 2 diabetes patients in community of Jiading district of Shanghai
Min-Qian SHEN ; Hua YANG ; Yuan-Zhi YUAN ; Shan-Zhu ZHU ; Jia-Fen WEI ; Fei YUAN
Chinese Journal of Clinical Medicine 2017;24(5):723-727
Objective:To investigate the prevalence and related risk factors of diabetic retinopathy (DR) among registered residents of type 2 diabetes mellitus in Jiading Industrial Zone Community Health Service Center,Shanghai.Methods:In this cross-sectional study,diabetic retinopathy was screened among registered residents with type 2 diabetes in Jiading Industrial Zone Community Health Service Center,Shanghai.The prevalence and risk factors of DR were evaluated by combining questionnaire and clinical examination.Results:Among 776 subjects who were chosen,695 people completed the study,and the overall response rate was 89.56%.26 cases were excluded because the refractive medium was opaque and the retina could not be seen clearly.DR was detected in 91 patients (13.60%).The mean duration of diabetes in patients with DR and without DR was (9.63±6.44) years and (7.02±5.32) years (OR=1.073,95% CI 1.036-1.111,P<0.001),the fasting blood glucose was (8.66±2.66) mmol/L and (7.52±2.15) mmol/L (OR=1.219,95% CI 1.119-1.328,P<0.001),glycated hemoglobin was (7.68±1.55)% and (6.99±1.21)% (OR=1.495,95% CI 1.275-1.754,P<0.001).There was also statistically significant difference in the use of insulin between DR patients and non-DR patients (OR=4.202,95 % CI 2.504-7.051,P<0.001).Conclusions:The prevalence of DR among type 2 diabetes patients is 13.6% in Jiading Industrial Zone,Shanghai.The longer the duration of diabetes is,the higher the fasting blood glucose is,the higher the glycosylated hemoglobin is,the greater the risk of DR will be.
6.Procollagen type 1 N-terminal propeptide and beta-collagen special sequence for the diagnosis and treatment of osteoporosis
Jun ZOU ; Chenxi YUAN ; Hongjun ZHU ; Ying LU ; Min LIN ; Yijia CHEN ; Junhua WANG ; Jiaxuan XU
Chinese Journal of Tissue Engineering Research 2015;(29):4731-4735
BACKGROUND:Procolagen type 1 N-terminal propeptide (P1NP) and β-colagen special sequence(β-CrossLaps) are two bone metabolic markers that are closely related to osteoporosis. Combined detection of bone metabolic markers and bone mineral density is of clinical significance for the diagnosis of osteoporosis. Bone metabolic markers are ideal indicators to predict fractures, which can compensate for the lack of bone density test. OBJECTIVE:To introduce the application of bone metabolic markers in the monitoring of drug efficacy on the treatment of osteoporosis as wel as in the prediction of fracture risks in recent 20 years and to explore the clinical values of P1NP and β-CrossLaps to assess the therapeutic efficacy on osteoporosis and risks for osteoporotic fractures. METHODS:A computer-based search of CNKI and SCI databases were performed for relevant articles published from 2000 to 2014 using the keywords of “serum bone metabolic markers; osteoporosis; bone mineral density” in Chinese and English, respectively. Finaly, 44 articles meeting the inclusive criteria were reviewed. RESULTS AND CONCLUSION:This paper analyzes the source and detection mechanisms of P1NPand β-CrossLaps and then compares their advantages in the therapeutic effect assessment of osteoporosis. Serum bone metabolic markers cannot only reflect the dynamic changes of bone metabolism, but also have earlier changes than the bone mineral density. Both P1NPand β-CrossLaps are very important for assessing the early diagnosis of osteoporosis as wel as anti-osteoporosis drug efficacy.
7.The role of diffusion-weighted imaging for breast MRI
Qinghua MIN ; Jun YANG ; Kangwei SHAO ; Caisong ZHU ; Lixin YUAN ; Huiling XU ; Peng LI ; Wenjie LU
China Oncology 2015;(8):602-607
Background and purpose:Diffusion-weighted imaging (DWI) is a non-invasive technique of breast magnetic resonance imaging (MRI). DWI is an alternative to dynamic contrast-enhanced (DCE) MRI for differentiating malignant from benign lesions in breast screening or not. This study aimed to evaluate the potential role of DWI in differentiating malignant breast lesions from benign lesions.Methods:Seventy-four patients underwent digital mammography, DCE and DWI (49 patients’b-value of 0, 400, 600 and 800 s/mm2). The detectability, sensitivity and speciifcity of DWI and DCE were compared. Absolute apparent diffusion coefifcient (ADC) was compared with standardized ADC for quantitative analysis.Results:Sixty-four of 74 patients had positive pathologic findings (38 malignant, 26 benign). All of the malignant lesions were detected on DWI and DCE. The sensitivity of DWI was 83.33%, 90.00% and 93.33%, and the specificity was 85.91%, 76.19% and 72.72%, forb-value of 400, 600 and 800 s/mm2, respectively. The sensitivity and speciifcity of DCE were 86.61% and 90.48%. There was no signiifcant difference between absolute and standardized ADC in detecting breast cancer (P>0.05).Conclusion:DWI is an important complemented technique to DCE-MRI for differentiating malignant from benign lesions in breast MRI.
8.Prevalence and clinical features of human metapneumovirus infection in hospitalized children with lower respiratory tract infection
Mei ZENG ; Quan LU ; Yuan QIAN ; Ru-Nan ZHU ; Min LU ; Hui-Yan ZHANG ;
Chinese Journal of Infection and Chemotherapy 2006;0(04):-
Objective To elucidate the prevalence and clinical characteristics of human metapneumovirus(hMPV)in hospital- ized children with respiratory infection.Methods A total of 452 hospitalized children with lower respiratory tract infection were observed from Aug 2004 to Jan 2005.Respiratory tract aspirates were collected from all patients within 48 hours after admis sion.The specimens were routinely tested for respiratory syncytial virus,influenza virus A and B,parainfluenza virus 1 to 3 and adenovirus by direct fluorescent assay(DFA).The 245 specimens negative by DFA were tested for hMPV by RT-PCR. PCR products of hMPV M gene from some patients were randomly selected for sequencing analysis.Results hMPV was identi- fied in 59(24.1%)of the 245 specimens tested,hMPV infection alone accounted for 13.1% of the infections in the 452 chil- dren under study,The prevalence of hMPV was higher than other respiratory viruses in winter.The mean age of hMPV-infec- ted children(n=59)was 27.7 months.There was no significant difference between age groups in terms of the prevalence of hMPV(P>0.05).There were no statistically significant difference in demographics and clinical symptoms between hMPV in- fection and other common respiratory virus infection.Genotyping for the hMPV M gene from 23 Shanghai patients showed two distinct hMPV genotypes.Sequence analysis of these hMPV M genes showed 82.8%-100% homology to the registered se- quence in GenBank.There was no significant difference in clinical characteristics between the 2 genotypes.Conclusions hMPV plays an important pathogenic role in lower respiratory tract infection of children,hMPV prevailed in the winter of 2004.Clini- cally,hMPV infection can not be discriminated from the infection of other respiratory viruses.Clinical manifestation is similar between the two hMPV genotypes.
9.Investigation of PML-RAR? fusion gene in acute promyelocytic leukemia by real-time quantitative reverse transcription PCR
Hong YUAN ; Nan WANG ; Shi-Jun LI ; Jing WANG ; Hong ZHU ; Xiao-Guang XIAO ; Min HUANG ;
Chinese Journal of Laboratory Medicine 2001;0(04):-
Objective To investigate the relapse risk assessment for patients with acute promyelocytic leukemia(APL)through testing PML-RAR? fusion gene by real-time quantitative polymerase chain reaction(RQ-PCR).Methods Relative copies of PML-RAR? fusion gene were measured in 25 patients with APL in phases of first diagnosis,complete remission(CR)and relapse.The minimal residual disease(MRD)situations were also monitored in 6 of the 25 patients.Results Different PML-RAR? fusion gene expression levels were observed in patient groups of different phases of the disease.(P
10.The Research and Practice of Running School Pattern in Local Medical Colleges and Universities
Youai DU ; Linbo YUAN ; Shunqin MIN ; Xuebo ZHU ; Dongwu XU ; Pengpeng JIN
Chinese Journal of Medical Education Research 2003;0(04):-
Local medical college is the mainstream of altitude medical college education in our country.Facing furious competition,we must make a reasonable orientation,construct the characteristics of running school,establish scientific concept,construct specific subject and specialty and make an all-around development in order to walk up to national stage by depending on powerful unity,serve the regional economy and social development,and form a new situation of two patterns by converting single medical colleges to all-around college and remaining the existing single medical college.