Objective To establish a method of isolating and cultivating rat bone marrow mesenchymal stem cells (MSCs) in vitro and the biological characteristics thereof.Methods MSCs were isolated and purified from the bone marrow of rat by density gradient centrifugation and by adhering to the culture plastic.After successive subculture and amplification,the growth curve was drawn.The morphology of MSCs was observed in primary and passage culture under inverted microscope.The cells surface antigens of CD29,CD38,CD44 and CD45 were detected through flow cytometer.Results Cultured primary MSCs were spin dle-shaped and had a typical fibroblast-like morphology,proliferated in colonies.After a series of passage,the attached cells became morphological homogeneous and proliferated quickly.The growth curves of passage 1,3 and 5 were quite similar.The surface marker identification showed that CD44 and CD29 were positive,but CD45 andCD38 were negative.Conclusion The MSCs can be successfully isolated,cultured and purified from rat bone marrow through density gradient centrifugation,adherent property and subculture.MSCs can greatly proliferate after a short period of culture in vitro.The MSCs can be used as a potential cell source for tissue engineering.

AIM: To investigate the effect of antisense oligodeoxynucleotides against platelet-derived growth factor B-chain(PDGF-B) in the proliferation of cultured rat hepatic stellate cells and collagen-Ⅰ production.METHODS: The phosphorothioate antisense oligodeoxynucleotides and missense oligodeoxynucleotides were synthesized.The rat hepatic stellate cells(HSCs) were treated with these oligodeoxynucleotides at different concentrations,respectively.The inhibition of the proliferating of cultured rat hepatic stellate cells was assayed by MTT method.The expressions of PDGF-B and collagenⅠ mRNA were detected by means of reverse transcription-polymerase chain reaction(RT-PCR).The expression of endogenous PDGF-B was determined by flow cytometry(FCM).Collagen-Ⅰin the supernatant was determined by ELISA.RESULTS: The results showed that the ASODN against the PDGF-B at a final concentration of 10 ?mol/L inhibited the HSCs proliferation,the expressions of PDGF-B and collagen-ⅠmRNA.FCM and ELISA demonstrated that the expression of endogenous PDGF-B and the production of collagen-Ⅰin HSCs were significantly lower than those in controls,whereas the controls(missense oligodeoxynucleotides) had no effect at the same concentration.CONCLUSION:PDGF-B ASODN inhibits the proliferation of cultured rat hepatic stellate cells,the endogenous expression of PDGF-B and the production of collagen.PDGF-B ASON might be useful for gene therapy in liver fibrosis.

Renal Tubular Cell Apoptosis refers to activation,proliferation,distension,cast formation and cellular apoptosis of the tubular epithelium when the renal tubule undergoes hypoxia,poisoning,inflammatory reaction,plasma protein or amylaceum.This text summarized the researches on the prevention and control of lenal tubular cell apoptosts in renal interstitial fibrosis in the recent years.

Aim To demonstrate the effects and mechanism of adenosine A1 receptor agonist R(-)-N6-(2-phenylisopropyl) adenosine(R-PIA) on high glucose(HG)-induced myocardial hypertrophy by in vitro cultured myocardial cells from neonatal rats.Methods The protein content was assayed by the method of Lowry. The expression of p-ERK1/2 and ERK1/2 was determined by Western blot.The [Ca~(2+)]I transient changes of cell loaded Fura-2/AM were measured by Till image system.Results 1 μmol·L~(-1) R-PIA and U0126 inhibited similarly HG-induced increase of the protein content and [Ca~(2+)]I transient along with the relative expression of p-ERK1/2.These responses were completely abolished by adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine(CPDPX).Conclusion Adenosine A1 receptor stimulation significantly inhibits HG-induced myocardial hypertrophy by mediating ERK1/2 pathway and Ca~(2+).

Cytokines is a kind of important factor on Pathogenic of hysteromyoma.Some study provides evidence that sex homone and its product of metabolism react up regulation/down regulation for hysteromyoma by growth factor. Apoptosis and oncogene and retinoid X have influence to the growth of hysteromyoma.

Objective To evaluate the labor duration of healthy primiparas in the past 20 years compared with Friedman labor curve.Methods Published observational studies about labor duration in primiparas with singleton vertex presentation were searched in PubMed,EMBase,Cochrane library,CNKI,VIP and Wanfang database.The Cochrane Collaborafion' s RevMan 5.1 software was used for reta-analysis.Results Eleven literatures involving 4534 primiparas were included,which were eligible for the criteria to investigate the labor duration.Meta-analysis showed that the length of active phase was significantly different between what was showed in primiparas in the past 20 years and Friedman labor curve[ P =0.01,weighted mean difference ( WMD ) =1.61,95％ CI:0.38 to 2.83 ],and significant differences were also found in the second stage duration (P =0.0006,WMD =-0.20,95％ CI:-0.31 to -0.09 ).Conclusions Compared with Friedman labor curve,active phase length of healthy primiparas in the past 20 years was significantly longer and second stage length was shorter.Reassessing labor curve of healthy primiparas is required for more scientific guidance and less unnecessary interventions.

Objective To construct a highly expresses catalase of Helicobacter pylori (H.pylori) and to test it's activity. Methods The catalase DNA was amplified from H.pylori chromosomal DNA by PCR techniques, inserted into the prokaryotie expression vector pET 22b(+) and then transformed into the BL21(DE3) E.coli strain which expressed catalase recombinant protein. Then the activity of H.pylori catalase was assayed by the Beers and Sizers. Results DNA sequence analysis showed that the sequence of catalase DNA was the same as that published by GenBank. The catalase recombinant protein amounted to 24.4% of the total bacterial protein after inducing with IPTG for 3 hours at 37?C and the activity of H.pylori catalase was high in the BL21 (DE3) E.coli strain. Conclusions A clone of expressing high activity H.pylori catalase has been obtained, and thus a good foundation for further study has been laid.

Objective To evaluate the high-risk human papillomavirus(HR-HPV)detection and thinprep cell test(TCT)in the diagnosis of cervical lesion by comparison with pathological findings.Methods The data collected from Dec.2004 to Dec.2006 from 690 patients who had undergone HR-HPV detection,TCT and pathology examination with electronic colposcopy in the Department of Obstetrics and Gynecology,General Hospital of PLA were analyzed retraspactively.Results Among the 690 patients,the coincidence between the findings of TCT and pathology was 22.34%(42/188)for low-grade squamous intraepithelial lesion(LSIL),58.33%(56/96)for high-grade squamous intraepithelial lesion(HSIL)and 100%(16/16)for cervical cancer.The number of patients with the cytological diagnosis of abnormal squamous cells(ASC),LSIL,HSIL and cervical cancer was 315,188,96 and 16,respectively.Among them,the positive rate of HR-HPV of ASC,LSIL,HSIL and cervical cancer was 53.96%(170/315),77.12%(134/188),80.21%(77/96)and 100%(16/16),respectively.Pathologic examination through electronic colposcopy revealed the number of patients with the diagnosis of inflammation,CINⅠ/HPV,CINⅡ/CINⅢ and invasive cancer was 425,81,157 and 27,respectively.Among them,the positive rate of HR-HPV in inflammation was 43.29%(184/425),CINⅠ/HPV 74.93%(60/81),CINⅡ/CINⅢ 91.72%(144/157)and invasive cancer 92.5%(25/27).Among the 690 cases,413 were HR-HPV positive,55.45%(229/413)of which showed CINⅠ/HPV or higher.277 cases were HR-HPV negative,and 87.01%(241/277)of which showed inflammation,12.99%(36/277)of which showed CINⅠ/HPV or higher,and 5.42%(15/277)of which showed CINⅡ/CINⅢ and invasive cervical cancer.Among 315 cases with ASC,170 cases were HR-HPV positive,41.76%(71/170)of which showed CINⅠ/HPV or higher,including 69.01%(49/71)CINⅡ/CINⅢ and invasive cervical cancer.145 cases were HR-HPV negative,only 4.83%(7/145)of which was CINⅡ/CINⅢ and invasive cervical cancer,and 90.34%(131/145)of which was inflammation.Conclusion HR-HPV detection and TCT are feasible for cervical lesion screening.Cytology combined HR-HPV test is favorable for shunting management in cervical lesions.

Objective To discuss the clinical significance of human papilloma virus (HPV) infection in cervical cancer and to explore the feasibility of detecting the infection with microarray from paraffin-embedded specimens of cervical cancer. Methods From May 2005 to February 2007, 48 patients with cervical carcinoma, including 37 cases with cervical squamous cell carcinoma and 11 cases with cervical adenocarcinoma, were analyzed retrospectively. After DNA extraction from the paraffin-embedded tissues, 23 HPV subtypes were detected by the use of microarray after amplification by polymerase chain reaction (PCR) and hybridization. Results 44 cases of cervical cancer were found to be high-risk HPV positive genotypes. The HPV infection rate was 91.7%. The HPV infection rate in cervical squamous cell carcinoma was 94.6% (35/37), and in cervical adenocarcinoma was 81.8% (9/11). Among them, 33 cases were found to have single infection, accounting for 75.0% of the infection rate. There were 11 cases of mixed infection, among them 9 cases were found to have double infection and 2 cases with multiple infection, accounting for 20.5% and 4.6%, respectively, of the infection rate. The infection rate of HPV16, the main genotype, was 90.9% (40/44). The infection rate of HPV18, the second ranking subtype, was 27.3% (12/44). Infection with HPV52, 33, 59, and 68 genotypes was lower in incidence. Among 35 cases of cervical squamous cell carcinoma, the infection rates of HPV16 and HPV18 were 91.4% (32/35) and 22.9% (8/35), respectively. Among 9 cases of cervical adenocarcinoma, the infection rate of HPV16 and HPV18 were 88.9% (8/9) and 44.4% (4/9), respectively. Conclusion Multiple HPV genotypes can be detected from paraffin-embedded tissues with microarray technique in high sensitivity and specificity, and it is useful to study the pathogenesis and prevention of cervical cancer.

ObjectiveTo investigate the practical value of non-invasive positive pressure ventilation (NIPPV) in the treatment of acute left heart failure(ALHF).MethodsThe clinical effects and complications of NIPPV performed on 36 cases with severe ALHF on the basis of conventional treatment were evaluated and compared with previous treatment without NIPPV.ResultsOf 36 cases with ALHF treated by NIPPV,30 cases improved,6 cases were altered to receive invasive ventilation or dead.Total effective rate was 83.3%.Compared with 42 cases with severe ALHF treated only by conventional treatment in the past 5 years,with only 26 cases being successfully salvaged and effective rate being 61.9%,significant difference (P