Objective To evaluate the role of hippocampal histone acetylation in isoflurane-induced amnestic effect in mice.Methods Fifty-four male C57BL/6J mice,aged 8 weeks,weighing 18-22 g,were randomly divided into 3 groups (n =18 each) using a random number table:control group (group C),isoflurane group (group ISO) and histone deacetylase inhibitor sodium butyrate group (group SB).Group C inhaled 35％ oxygen for 30 ain,and ISO and SB groups inhaled the mixture of 35 ％ oxygen and 0.4％ isoflurane for 30 min,and then the animals underwent contextual fear conditioning training.After the end of training,normal saline 6 ml/kg was intraperitoneally injected in C and ISO groups,while in group SB,sodium butyrate 1.2 g/kg was intraperitoneally injected.One hour after the end of training,3 mice were sacrificed randomly in each group and their hippocampi were immediately removed for determination of the expression of acetylated histone-H3 (Ac-H3) and Ac-H4 by Western blot.Twenty-four hours after the end of training,contextual fear conditioning test and open field test were conducted.The freezing time,total distance and time of staying at the central zone were recorded.Results Compared with group C,Ac-H3 and Ac-H4 expression was significantly down-regulated,and the percentage of freezing time during testing was decreased in group ISO (P ＜ 0.05).Compared with group ISO,Ac-H3 and Ac-H4 expression was significantly up-regulated,and the percentage of freezing time during testing was increased in group SB (P ＜ 0.05).There was no significant difference in the percentage of freezing time during training,total distance and time of staying in the central zone among the 3 groups (P ＞ 0.05).Conclusion Hippocampal histone acetylation is involved in the regulation of isoflurane-induced amnestic effect in mice.

BACKGROUND:Because of convenient source, multi-lineage differentiation and low immunogenicity, bone marrow mesenchymal stem cels are the ideal cel type to serve as vectors of transgenic cels in pain management. However, the replicative senescence and smal amount of cels obtained from the bone marrow restrict the application of bone marrow mesenchymal stem cels in pain research. OBJECTIVE:To construct human telomerase reverse transcriptase (hTERT)-immortalized rat bone marrow mesenchymal stem cels as transgenic celular vectors for pain therapy. METHODS:Bone marrow mesenchymal stem cels were obtained from whole rat bone marrow, and then transfected with a lentivirus containing the hTERT (pLV-Puro-EF1α-hTERT) folowed by puromycin selection. hTERT expression and telomerase activity in these transfected cels were determined by RT-PCR and TRAP. Morphological changes, capacity of cel growth and multi-lineage differentiation, chromosome karyotype and tumorigenicity were observed in vitro. Moreover, the expression of cel surface molecule, Nestin, MHC-I and MHC-II in transfected cels were also detected by flow cytometry and immunocytochemistry. RESULTS AND CONCLUSION: The bone marrow mesenchymal stem cels geneticaly modified by hTERT could be cultured and passaged through 30 generations in vitro. Compared to the primary and negative transfected cels, the hTERT-modified bone marrow mesenchymal stem cels showed higher expression of hTERT mRNA, telomerase activity and cel proliferation. Most of transfected cels stayed at G2/M and S stages. The proliferation index of the transfected cels were increased dramaticaly. The positive rates of CD29, CD44 and CD90 were over 70%, but the positive rates of CD34 and CD45 were less than 5%. Transfected cels were positive for Nestin in the cytoplasm, but negative for MHC-1 and MHC-11. In addition, this cel line continued to exhibit the characteristics of fibroblastic bone marrow mesenchymal stem cels, including phenotype, differentiation into osteoblasts, adipocytes and neuron-like cels. No chromosome abnormality and tumor formation were observed in this experiment. Taken together, these data suggests that the rat bone marrow mesenchymal stem cels immortalized by hTERT gene are constructed successfuly and stil maintain major stem cels characteristics, which provide safe and stable cel vectors as research base for pain therapy.

Objective To investigate the in vitro effects of acitretin on the apoptosis and expressions of insulin-like growth factor binding protein 7 (IGFBP7) and vascular endothelial growth factor (VEGF) in HaCaT cells.Methods Cultured HaCaT cells were treated with various concentrations (10-5,10-64,10-7,10-8 mol/L) of acitretin for various durations,with those cultured in acitretin-free medium serving as the control group.Then,CCK-8 assay was performed to evaluate the proliferation of cells after 24-,48-and 72-hour treatment,flow cytometry to detect the apoptosis of HaCaT cells,and Western blot and reverse transcription-PCR to quantify the protein and mRNA expressions of IGFBP7 and VEGF in HaCaT cells,respectively,after 48-hour treatment.Statistical analysis was carried out by one-way analysis of variance and Pearson correlation analysis.Results The proliferation of HaCaT cells was inhibited by the treatment with acitretin,and the inhibitory effect increased with the elevation of concentration and prolongation of treatment duration of acitretin.A significant decrease was observed in the proliferative activity of HaCaT cells treated with acitretin of 10-8 mol/L for 48 hours,and when the concentration of acitretin was 10-5 mol/L,the proliferation of HaCaT cells was inhibited by 39.94％ ± 2.27％ and 49.77％ ± 1.87％ at 48 and 72 hours respectively,compared with the control cells.The HaCaT cells treated with acitretin of 10-5 mol/L for 48 hours showed a significant elevation in apoptosis rate (7.617％ ± 0.767％ vs.1.803％ ± 0.313％,P ＜ 0.05),IGFBP7 protein and mRNA expressions (0.939 ± 0.040 vs.0.436 ± 0.013,0.872 ± 0.079 vs.0.190 ± 0.056,both P ＜ 0.05),but a significant reduction in VEGF protein and mRNA expressions (0.213 ± 0.032 vs.0.798 ± 0.036,0.274 ± 0.041 vs.0.933 ± 0.054,both P ＜ 0.05) in comparison to the control cells.Conclusions Acitretin can induce the apoptosis of HaCaT cells,and up-regulate IGFBP7 but down-regulate VEGF expressions in HaCaT cells at protein and mRNA levels.

OBJECTIVE: To investigate the clinical features of chronic tracheoesophageal fistula (TEF), provided disease-related treatment experience and lessons for clinicians. METHOD: To successfully repair one case of chronic tracheoesophageal fistula with surgery, and to analyze the clinical treatment process, combined with relevant literature, the author reported the experiene of diagnosis and treatment in TEF. RESULT: After the gastrointestinal ostomy and Stent implantation, the fistula persisted, nine months later ,we took the surgery to repair the fistule, ten days postoperation, the fistule healed and esophageal iodine water examination didn't prompt obvious abnormalities, the patient was discharged without any postoperative complications 12 days postoperation. CONCLUSION If conservative treatment failed with TEF, the surgical repair should be carried timely, By double sutured with fistula in surgery, and reinforced with the approaching muscle tissue, It can achieve good results.
Humans ; Postoperative Complications ; Postoperative Period ; Tracheoesophageal Fistula ; pathology ; surgery ; Wound Healing

Objective To investigate the prevalence of adult osteofluomsis in the endemic fluomsis areas in Tianjin and to provide scientific foundation for endemic fluorosis.Methods Stratified sampling in 55 villages were selected in 3 areas with slight,moderate and severe fluorosis regions in Tianjin from April to June in 2008.Water fluorine were tested and clinical osteofluorosis examinations were conducted to the population aging 16 and above in the villages.Tweenty villages were selected randomly in the slight,moderate and severe fluorosis regions.X-ray osteofluorosis examination were conducted to patients and suspected patients in these 20 villages.Results The geometric mean fluoride content in the water for the 3 areas were 1.35 mg/L,3.44 mg/L,5.49 mg/L,respectively.The prevalence of osteofluorosis were 36.7%(44/120),20.6%(33/160),39.4%(43/109),respectively.The prevalence of osteofluorosis Was increased gradually(r=0.534,P<0.01)and the symptoms and signs of the disease were more serious(H=84.813,P<0.01).The prevalence of X-ray diagnosis Was increased gradually(r=0.990,P<0.01)and signs of the disease were more severe(H=25.169,P<0.01)with an increase in age.There was no statistical significance of prevalence rate of osteofluorosis between males and females,regardless if it Was a clinical diagnosis(X2=0.343,P>0.05)or an X-ray diagnosis(X2=3.532,P>0.05).Conclusions Adult osteofluorosis to a certain extent is still prevalent in the fluorosis areas in Tianjin.Endemic fluorosis is still rampant.Improving water in fulorosis areas should be mandatory.

Objective To explore the feasibility of extending liver blood perfusion cessation by ultrasound combining microbubbles.Methods The livers of 9 healthy rabbits were treated with a pulsed therapeutic ultrasound device,in presence of microbubbles.For quantification of liver perfusion,contrastenhanced ultrasonography was performed on 6 rabbits before treatment and at different time points of 0 min,30 min,60 min and 48 hours after treatment.Pathological examination of the treated livers was performed immediately after treatment on the other 3 rabbits.Results The liver blood perfusion almost vanished immediately after treatment,remained at a low perfusion level from 30 to 60 min,and completely recovered 48 hours later.The peak intensity dropped from ( - 51.88 ± 4.26)dB to ( - 62.53 ± 4.83)dB after treatment and rose up to ( - 52.00 ± 4.60) dB 48 hours later.The peak intensities before treatment and 48 hours after treatment were significantly higher than those of 0 min,30 min and 60 min time points after treatment( P ＜0.05).Pathological examination showed significant swelling of hepatocytes and hemorrhage around portal veins.Conclusions Microbubbles enhanced ultrasound can induce liver blood perfusion cessation for up to 1 hours.The mechanism could be swelling of hepatocytes and hemorrhage of portal track.

Objective To compare the therapeutic effects of bone marrow stem treatment by different bone marrow mobilization,distinct separation methods or cell suspension density, and to explore the optimal treatment plan of autologous bone marrow stem cell transplantation for decompensated cirrhosis.Methods Twenty three patients with decompensated cirrhosis were studied.100 ~200 mL bone marrow from each patient was harvested in aseptic condition, after isolation and purification by density gradient centrifugation,the stem cells were obtained and transplanted into the liver via hepatic artery.The serum glutamic acid alanine aminotransferase (ALT), albumin (ALB), prothrombin time (PT) and total bilirubin (TBIL) were checked before and l, 2, 3 months after therapy respectively,changes in these indicators of each group were compared by different bone marrow mobilization, distinct separation methods, or cell suspension density.Results After transplantation, levels of albumin increased significantly 1, 2, 3 months after treatment compared with baseline(P<0.05).Patients in bone marrow mobilization group obtained higher stem cell density (P<0.05), which had no significant difference in improving liver function ( ALT, TBIL, ALB, PT) compared with non-bone marrow mobilization group.Patients using the kit significantly improved their albumin (3 months after treatment, P<0.05), which had no significant difference in improving liver function (ALT, TBIL, ALB, PT) compared with using ordinary lymphocyte isolation method group.There was no significant difference in improving liver function ( ALT, TBIL, ALB, PT) between lower magnitude stem cell density group(≤1 ×1010/L) and higher magnitude stem cell density group(1 ×1010/L).Conclusion The treatment for decompensated cirrhosis by transplantation of autologous bone nlarrow stem cells is safe and effective, which can significantly increase the level of albumin in patients with decompensated cirrhosis.Preoperative bone marrow mobilization can increase the rate of stem cells obtained, and the method using the kit improves the leves of protein respectively.They are helpful to improve the efficacy.

Objective To examine the occupational stability,training needs and capacity building requirements of the general practitioners purposely trained to work in rural communities,for improvement of policies regarding their training and issues related. Methods There counties were selected by stratified random sampling, where general practitioners graduated from such training completed an anonymous questionnaire. Focus group interviews were made to officials of the local Health and Family Planning Commission,those of primary healthcare institutions,and general practitioners themselves. Results The of the 190 valid questionnaires recovered, male accounted for 34. 2% and female 65. 8%. These general practitioners were mostly satisfied with their working environment, social status, work stress and job achievement,yet with lower satisfaction with their salary and availability of medical devices. Consideration of resignation was reported by only 5. 3% of them,with more males than females. As high as 63. 2% of them prefer to work in hospitals of county level. 91. 1% of them held it as necessary to participate in standard resident training,and 98. 4% held it as necessary to have on-job training. 94. 7% of them have participated in further education to gain a bachelor degree in medicine. Conclusions These doctors were found with high stability to work in rural communities and low turnover intention. Further studies are required to encourage these general practitioners to serve rural communities in a long term, and to improve the standard resident training mechanism,and their career development.

Objective To optimize and simplify the survey method of Oncomelania hupensis snail in marshland endemic re?gion of schistosomiasis and increase the precision,efficiency and economy of the snail survey. Methods A quadrate experimen?tal field was selected as the subject of 50 m×50 m size in Chayegang marshland near Henghu farm in the Poyang Lake region and a whole?covered method was adopted to survey the snails. The simple random sampling,systematic sampling and stratified ran?dom sampling methods were applied to calculate the minimum sample size,relative sampling error and absolute sampling error. Results The minimum sample sizes of the simple random sampling,systematic sampling and stratified random sampling meth?ods were 300,300 and 225,respectively. The relative sampling errors of three methods were all less than 15%. The absolute sampling errors were 0.221 7,0.302 4 and 0.047 8,respectively. Conclusion The spatial stratified sampling with altitude as the stratum variable is an efficient approach of lower cost and higher precision for the snail survey.

AIM To study the effects of coadministration of berberine chloride(Ber) with cyclosporin (CsA) on liver microsomal cytochrome P450 isoenzyme and multi drug resistance gene in rats. METHODS The activities of liver microsomal erythromycin demethylase(ERD) and aminopyrence N demethylase(ADM) were determined. The levels of mRNA expression of CYP3A1, CYP1A1, CYP2E1, mdr1a and mdr1b were assayed with RT PCR. RESULTS After administration for 6 days, all treatment groups except Ber at 100 mg?kg -1 exhibited inhibitory action on ERD activity in rats. ERD activity markedly decreased in all drug treatment groups after taking drug for 12 days. After administration for 6 days, 45 mg?kg -1 CsA, 100 mg?kg -1 Ber coadministrated with 45 mg?kg -1 CsA and 200 mg?kg -1 Ber plus 45 mg?kg -1 CsA had significant inhibitory effects on ADM activity in rats. All groups except 100 mg?kg -1 Ber and 150 mg?kg -1 ketoconazole had the same effects on ADM activity after treatment for 12 days. Again, after 12 days, all drug treatment groups except 100 mg?kg -1 Ber group was found of remarkable inhibition of the mRNA expression of CYP3A1, CYP2E1, mdr1a and mdr1b. The CYP1A1 gene was not detected in all groups. CONCLUSION The mechanism of Ber to increase CsA concentration is that Ber decreases the expression of CYP3A, mdr1a and mdr1b thereby reduces the metabolism and elimination of CsA by liver.