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.

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

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.

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.

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.

Objective To track the effects of hyperbaric oxygen (HBO) therapy on neonatal rats with hypoxic-ischernic brain damage (HIBD).MethodsA total of 126 healthy,seven-day-old SD rats were used to model the HIBD and then randomly divided into seven groups of 18:a sham group,a hypoxic-ischemic (HI) group,a 6 h HBO group,a 24 h HBO group,a 48 h HBO group,a 72 h HBO group and a 1 week HBO group.One hour of HBO treatment at 2 atmospheres absolute pressure was administered once daily for a week to the rats in the latter 5groups,starting at 6,24,48,72 hours and 1 week post the HIBD modeling,while no HBO was administered to the sham and HI groups.The effects were assessed in terms of histological changes ( the neuron density and the percentage of neuron apoptosis in the CA1 region of the hippocampus) and nitric oxide (NO),malondialdehyde (MDA) and superoxygen dismustase (SOD) concentrations after 15 days.ResultsIn the 6 h HBO,24 h HBO,48 h HBO and 72 h HBO groups,average neuron density in the CA1 region was significantly higher than in the HI group.But in the 1 week HBO group the average density was not significantly different than in the HI group.In the 6 h HBO,24 h HBO,48 h HBO and 72 h HBO groups the average percentage of neuron cell apoptosis in the CA1 area of the hippocampus was significantly lower than in the HI group,but the 1 week HBO group again showed no significant difference.There were significant differences in average NO,MDA or SOD levels among the 6 h HBO,24 h HBO,48 h HBO,72 h HBO and HI groups,but the 1 week HBO group showed no significantly higher average levels than the HI group.ConclusionsThe optimal therapeutic window for using HBO to protect against HIBD is within the first week.The best effect can be obtained in the first 6 hours,but after 1 week HBO no longer has a significant effect.The earlier the HBO is administered,the better the effect obtained.

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.

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

AIM: To explore a new compute tomography (CT) for the localization of intraocular foreign bodies (IOFBs).METHODS: After CT ocular horizontal and ocular axial scan,the foreign bodies on the eyeball wall or retinal surface in 26eyes were localized by means of the combination of CT ocular axial scan and meridian plane reconstruction (new method),ocular horizontal scan and ocular axial scan (conventional method Ⅰ) as well as ocular horizontal scan right angle coordination (conventional method Ⅱ) separately. According to the criteria of indirect ophthalmoscope localization and direct observation during operation, the relative accuracies of corresponding points of the foreign bodies on sclera surface along meridian and latitude were measured.RESULTS: In the 26 cases, the mean relative accuracies of corresponding points of foreign bodies on sclera surface along meridian and latitude were 1.53mm、1.64mm (new method), 1.37mm、1.64mm (conventional method Ⅰ) and 2.02mm、2.55mm (conventional method Ⅱ) respectively.There was no statistical difference between the new method and the conventional method Ⅰ, whereas there was statistical difference between the new methods and the conventional method Ⅱ (along meridian: P＜0.05; along latitude:P＜0.01).CONCLUSION: Compared with the conventional methods,the new method is simpler, more visible and more potential in the clinic.

ObjectiveTo analyze the prevalence and epidemic trend of human brucellosis between 2007 -2010 in Qian'an county,and to provide a basis for future prevention and control measures. Methods Epidemiological data of brucellosis between 2007 - 2010 were obtained from the national disease surveillance report on management information system, population data were from the national disease surveillance information management system for basic information reporting system, and descriptive epidemiological methods was used for statistical analysis. Results A total of 86 cases were infected with brucellosis in Qian'an county from 2007 to 2010, with 68 cases of male and 18 cases of female, and male to female ratio was 3.78 : 1. Patients were mainly in the age of 20 - 55, with 40 - ＜ 45 year-old group the highest. All patients were local residents. Most professional were livestock acquisition, processing and aquaculture personnel. The majority of patients lived in Jianchang town,with 51 cases, accounting for 59.30％. Thirteen patients were reported in 2007, incidence was 0.0201‰(13/ 647 983). Thirty-nine patients were reported in 2010, and incidence was 0.0563‰(39/657 380). There was a fluctuations increase from 2007 to 2010. In 2007 brucellosis occurred in only two townships(towns), which spread to eight townships (towns) and urban areas in 2010. ConclusionsThe epidemic of human brucellosis in Qian'an is in a spreading trend. We recommend the government to strengthen the quarantine of livestock, and the infected livestock should be timely treated. Strengthen the prevention and control in Jianchangying. At the same time increase the brucellosis propaganda, and enhance self-protection awareness of the occupational groups.