Objective To investigate the effect of tetramethylpyrazine pretreatment on the expression of cfos and heat shock protein (HSP70) during hypoxia-reoxygenation (H/R) in cultured fetal rat hippocampal neurons. Methods After the neurons were cultured and identified, they were randomly divided into 5 groups ( n = 24each): control group (group C), H/R group, and low, median and high concentration of tetramethylpyrazine pretreatment groups (group L, M and H). The neurons were exposed to 2 h of hypoxia followed by 24 h of reoxygenation. Tetramethylpyrazine was added with the final concentrations of 60, 200 and 800 μg/ml in group L, M and H respectively, and then the neurons were incubated for 1 h before H/R. The apoptosis rate, cell viability and expression of c-fos and HSP70 were detected. Results The cell viability was significantly lower, while the apoptosis rate was significantly higher in group A/R, L and H than in group C (P ＜0.01). The cell viability and HSP70expression were significantly higher, while the apoptosis rate and c-fos expression were significantly lower in group L, M and H than in group A/R, and in group M and H than in group L (P＜ 0.05). The cell viability and HSP70expression were significantly lower, while the apoptosis rate and c-fos expression were significantly higher in group H than in group M ( P ＜ 0.01 ).Conclusion The mechanism by which tetramethylpyrazine pretreatment inhibits the apoptosis in cultured fetal rat hippocampal neurons during H/R may be related to the dowm-regulation of c-fos expression and up-regulation of HSP70 expression.

Objective To investigate the apoptotic effect of microRNA-1 (miR-1) on hypoxemic cardiomyocytes. Methods The cultured H9C2 cells were divided into 5 groups:normal control group, negative control group, H2O2 group, miR-1 group and H2O2+miR-1 group. After verified the success of transfection by real time PCR, MTT and flow cytometry methods were used to test the cell vitality and apoptotic rate, while the mRNA and protein expression level of Bcl-2 were de-tected by real time PCR and Western blot methods. Results Compared with normal control group, there were no significant differences in all indexes in negative control group. The application of H2O2 and miR-1 respectively or together significantly increased the miR-1 level and apoptotic rate, and reduced the cell vitality and Bcl-2 expression level. Conclusion mi-croRNA-1 can induce cardiomyocyte apoptosis by downregulating anti-apoptosis factor Bcl-2.

Objective To investigate the protective function and mechanism of microRNA-1 (miR-1) downregula-tion in H2O2 injured cardiomyocytes. Methods The experiment was divided into 4 groups:Blank group, Negative Control group (NC), H2O2 group and H2O2+AS-miR-1 group. Cells in blank group were not underwent any treatment. Cells in NC group were transfected with random miRNA fragment. H2O2 and H2O2+AS-miR-1 groups were defined as H2O2 injured car-diomyocytes without or with transfected antisense miR-1 oligonucleotide(AS-miR-1). Real time PCR was used to test miR-1 transcription level, and cell vitality and apoptosis were analyzed by MTT and flow cytometry. The target gene of miR-1 was predicted by bioinformatics, then its mRNA transcription and protein expression level of Bcl-2 were detected by real time PCR and western blot respectively. Results There is no significant difference of all index between Blank group and NC group. H2O2 can induce cardiomyocyte injury, increase miR-1 level and rise apoptosis rate, reduce cell vitality and de-crease Bcl-2 expression level. Transfection of AS-miR-1 can decrease cell apoptosis, increase cell vitality and enhance Bcl-2 expression level. Conclusion Downregulation of microRNA-1 can protect cardiomyocytes that was injured by H2O2 through increasing anti-apoptosis factor Bcl-2 expression.

Objective To preliminarily analyze the credibility and validity of Chinese Medical Constitution Questionnaire(CMCQ).Methods CMCQ was used to investigate the distribution of Chinese medical constitution types of 450 women inpatients with postoperative nausea and vomiting (PONV) after gynecologic laparoscopic surgery from March to October of 2013.Credibility and validity were analyzed by Cronbach's α coefficient and factor analysis.Results Mixed constitution was identified in 184 (40.9％) patients.Cronbach's α coefficient of two dimensions (yang deficiency and qi stagnation) of CMCQ was proper,being 0.815 and 0.803 respectirely,while that of the dimension of harmony type was the lowest (0.514).The total variance explained by 9 common factors extracted from the results of factor analysis was only 48.5％.Conclusion CMCQ may need further revision based on the results of clinical application.

Objective To correct the high supratarsal fold after cosmetic upper blepharoplasty is a challenge because of the shortage of both upper eyelid skin and fat.Levator shortening has been a procedure of choice selected by the surgeons to correct this condition.Methods Under local anesthesia, the desired crease was marked according to the characteristics of eyelid anatomy and aesthetic consideration for Asian women.The incision was made through the marked new skin crease and the old incision with surrounding scar tissue was incised.The adhesion between the skin and the levator aponeurosis was completely released.The levator aponeurosis was then divided above the tarsal plate.It could be shortened and then reattached to the tarsal plate depending on the amount of the upper eyelid skin and the degree of the ptosis.The skin incision was then closed to form new but lowered fold.Results Between 2003 to 2015, a total of 34 Chinese women underwent bilateral levator shortening for correction of high supratarsal fold after previous unsatisfactory upper blepharoplasty by other surgeons.There were no surgical complications postoperatively in this series and 32 patients (94.1%) were satisfactory for the outcome of the proper height of the supratarsal fold with at least 1 month follow-up.Only one patient (2.9%) required surgical revision for asymmetry of supratarsal folds and one patient (2.9%) for correction of deepened supratarsal fold.Conclusions The levator shortening can be an effective procedure to correct high supratarsal fold after unsatisfactory upper blepharoplasty in Asian women.It is especially useful to correct such a condition where there is shortage of the eyelid skin combined with the eyelid ptosis.

Objective To develop the PCR-denaturing high performance liquid chromatography (DHPLC) for detection of E. Coli O157: H7. Methods The virulence genes of Shiga-like toxin(SLT) and rfbE were specifically amplified by 2 sets of primers. The target gene fragments of the PCR assay were 224 bp and 499 bp, respectively. Results Analysis of 37 strains demonstrated that this PCR system was specific. The detection limit of the PCR was 4 CFU/ml. Conclusion These results indicated that the multiplex PCR-DHPLC assay can be used for specific and sensitive detection of E. Coli O157:H7.

Objective To explore the effect of puncture the left or right branch of portal vein on the incidence of postoperative hepatic encephalopathy in transjugular intrahepatic portosystemic shunt (TIPS). Methods A literature search was conducted through investigating PubMed, Cochrane Library, CNKI digital library and Wanfang databases. Randomized controlled trials and cohort studies related to the effect of puncture the left or right branch of portal vein on the incidence of postoperative hepatic encephalopathy in TIPS were included in this review. According to the methods for Cochrane systematic review, the data were extracted, the quality of the literatures was evaluated, statistical analysis was conducted and the data were analyzed with RevMan 5.0. Results One randomized controlled trial and four cohort studies, including 552 patients with liver cirrhosis received TIPS, were finally included in this review. Random-effects model meta-analysis showed that puncture of the left branch of portal vein in TIPS caused lower incidence of postoperative hepatic encephalopathy than puncture of the right branch of portal vein (RR=0.51, 95%CI was 0.39 to 0.66, P<0.01). Conclusion Puncture of the left branch of portal vein in TIPS can significantly reduce the incidence of postoperative hepatic encephalopathy in comparison with puncture of the right branch of portal vein.

Objective: To observe the regulatory effects of miRNA-31 (miR-31) on LATS2 and cardiomyocyte hypertrophy via down-regulating miR-31 expression in rat's cardiomyocytesin vitro. Methods: Rat's cardiomyocytes were isolated and cultured for 10 daysin vitro, according to different intervention methods, the cells were divided into 4 groups:①Blank control group,②AngII intervention group,③Lentivirus with miR-31 inhibitor infection group,④Negative lentivirus infection group. On day-8, gene expressions of MiR-31, LATS2, cardiac hypertrophy ANP and β-MHC were examined by qRT-PCR; on day-10, cell morphology was observed by fluorescence staining. LATS2 protein expression was examined by Western blot analysis. Dual luciferase reporter plasmids were transfected into 293T cells, then luciferase activity was detected to identify the targeting effect of miR-31 on LATS2. Results: Compared with Blank control group, AngII intervention group showed increased gene expressions of miR31, cardiac hypertrophy ANP and β-MHC,P<0.05, enlarged cardiomyocyte surface,P<0.05; while decreased gene and proteinexpressions of LATS2,P<0.05. Compared with AngII intervention group, Lentivirus with miR-31 inhibitor infection group had down-regulated expressions of miR31, cardiac hypertrophy ANP and β-MHC,P<0.05, reduced cardiomyocyte surface, P<0.05; while slightly increased LATS2 gene expression and obviously increased protein expression,P<0.05. Dual luciferase reporter assay presented that relative luciferase activity of TRAF6-3' UTR+miR-146b was significantly decreased than TRAF6-3' UTR+miR-NC,P<0.01 and relative luciferase activity of LATS2-3' UTR+ miR-31 was signiifcantly reduced than LATS2-3' UTR-NC+miR-31,P<0.01. Conclusion: Cardiomyocytes hypertrophy could be reversed at certain degree by down-regulating miR-31; the targeting effect of miR-31 on LATS2 was involved in cardiomyocyte hypertrophyregulation.

Objective:To provide microsurgical anatomy data in the course, branch, distribution, arterial network profile of the submental artery and the range of the flap excision in submental flap transplantation.Methods:From March, 2015 to March, 2020, a total of 36 head and neck cast specimens were studied. Acrylic-butadience-styrene plastic (ABS) filler were perfused into the external carotid artery to make cast specimens. The course, branching, distribution and the arterial framework of the submental artery under a surgical microscope were investigated.Results:The submental artery originated from the facial artery before reaching the lower edge of the mandible (1.50±0.50) cm, with a diameter of (1.50±0.85) (0.6-2.3) mm. The main trunk of submental artery was (5.5±0.5) cm in length, which ran forward along the lower edge of the mandible and branched out (9.0±3.0) (7-13) branches with diameters between 0.1-0.5 mm, and mainly distributed to skin and superficial fascia of the submental area. The main trunk of submental artery divided into ascending, horizontal and descending branches about 3.0 cm of the midline of the mandible. The ascending branch went upwards over the lower edge of the mandible and joined up with the lower labial arch or participated in the formation of the lower labial arch; the horizontal branch divided into several branches and joined up with the branches from the opposite side; the descending branch branched posteriorly and inferiorly, joined up with branches of lingual artery and superior thyroid artery. The branches of the submental artery and the branches of the peripheral arteries were joined up in the submental area to form the submental artery network. The diameter of the vessels in the network ranged 0.1-0.2 mm. The arterial network was built in the form of 1 to 3 layers, and the area of main network was about 7.0 cm×5.0 cm.Conclusion:The submental artery has a long trunk, many branches and abundant anastomoses between the branches, forming a dense submental artery network, which provides sufficient pedicle length, rich blood supply and cutting area for submental flap. The flap can be transplanted free or transposed. The best location of submental flap is near the midline of arterial network, and the appropriate area is 7.0 cm×5.0 cm.

Based on literature review and on-site investigation, this paper analyzed the current policies, regulations, supply and demand dilemma and practical mode of such a combination, as well as the feasibility from the views of current policies, service offering and the practice. It is recommended to build a service system centering on " public health services + basic medical care + community elderly/medical support" , with services enriched and supportive mechanisms supplemented, thus transforming community health centers.