Objective To investigate the influence of hypoxia-reoxygenation on the myocardin gene expression of cultured rats' vascular smooth muscle cells(SMC).Methods The SMC were isolated from the media of the thoracic aorta vessel of Sprague-Dawley rats,and cultured with attachment-block manner.The morphology and cell counting of the cultured cells under hypoxia conditions were observed and comparedto that under normal culture condition.Total RNA extracted from the cultured cells,the expression of myocardin mRNA in SMC were measured at hypoxia status and at various time after reoxygenation through RT-PCR.Results The rats' vessel SMC was successfully cultured and showed a "peak-valley" shape.Self-made hypoxia equipment can produce a lower oxygen partial pressure without significant variation of pH value which met the experiment requirements in 2 4 hours .However,in the hypoxia conditions,the expression level of myocardin was lowest at the 12th hours,then increased.After 24 hours of hypoxia,the expression levels of myocardin began to increase at the 6th hour of reoxygenation and reached a normal level at the 12th hour of reoxygenation.Conclusions Hypoxia-reoxygenation has an effect on the expression of myodardin gene.

Objective To explore treatment of bile duct variation in Laparoscopic Cholecystectom.Methods The author retrospectively analyzed the clinical data of 7 cases with bile duct variation in 2 000 patients performing Laparoscopic Cholecystectomy.Among the 7 cases,2 cases had small hepatic duct openings in the gallbladder bed;2 cases had cystic duct openings in the right hepatic duct;2 cases had accessory right hepatic duct;and one case had rare variation whose right hepatic bile duct and the jejunum connect together.2 cases of the first variation had no bile leakage,adopting the suture method in LC.Among 2 cases of the second variation(all found in LC),one case had bile spillage in the junction of the cystic duct and the right hepatic duct,so the operator converses to laparotomy,cuts the gallbladder,sutures the break,and the patient had no bile leakage at last;The other one case was anatomized clearly under the cavity mirror.Among 2 cases of the third variation,one had no bile leakage,whose accessory hepatic duct was ligated in LC.The other one case had bile leakage after LC,so the operator converses to laparotomy,clips the accessory hepatic duct,and extract the drainage tube until there was no bile drainage.The last case was mistaken and cut it,the next day biliary peritonitis appeared,so the bile leakage was sewed up under the laparoscope.Results The seven cases were followed 1 ~3 years,they had no jaundice and their liver function was normal.Conclusion Careful-ly dissect Calot's triangle in LC,observe bile leakage after LC;improve the level of understanding and dealing bile duct variation in LC,don't cut the duct which is known to us.We should treat differently according to particular case.

Objective To explore the effects of wogonin on hyperlipidemia in mice and clarify the molecule mechanism. Methods Thirty mice were evenly divided into three group normal control group,model control group and treatment group. The normal control group was given normal diet,the model control group received high-fat diet,the treatment group received high-fat diet with wogonin (500 mg·kg-1 ). Results The mice developed hyperlipidemia 12 weeks after starting the high-fat diet. The body weight,visceral fat and fat index were increased (P<0. 05). After treatment,these indices were reduced ( P < 0. 01). Wogonin significantly reduced the total cholesterol ( TC),low density lipoprotein ( LDL),high density lipoprotein (HDL),except the triglyceride (TG). Compared to the model control group,the hepatic lipase(HL) and lipoprotein lipase(LPL) activity in the treatment group were recovered (P<0. 05),but HMG-CoA reductase activity was inhibited ( P<0. 01). Mechanistic study suggested that the lipid-lowering effect might be related to the lipid synthesis genes (SREBP-1c,FAS, PPARγ) and the lipid metabolism genes (PPARα,CPT-1). Conclusion Wogonin can prevent hyperlipidemia,which might be related to the regulation of enzyme activity and the changes of lipid synthesis and oxidative metabolism.

Objective: To study the effect of ghrelin on L-type calcium channel current (ICa-L) of ventricular myocytes in experimental rats. Methods: The single ventricular myocyte in experimental rats were obtained by enzymolysis method, and the whole-cell patch-clamp technique was used to investigate the effect of ghrelin on ICa-L of ventricular myocytes at different doses of 10 nmol/L, 100 nmol/L and 1μmol/L respectively. Results: Ghrelin at 10 nmol/L, 100 nmol/L and 1μmol/L may inhibit ICa-L at (8.95 ± 2.13) %, (31.18 ± 4.78) % and (64.63 ± 8.57)% respectively,P<0.05, and the current-voltage curve was shifted upwards. The channel half inactivation curve decreased from (-1.34 ± 1.9) mV to (-8.04 ± 1.32 ) mV, (9.76 ± 1.17) mV and (-11.81 ± 0.73) mV respectively,P<0.05, and the recovery time after inactivation was prolonged as τ value from (63.23 ± 9.32) to (98.95 ± 10.74), (109.56 ± 13.42) and (127.39 ± 16.13) respectively,P<0.05. Conclusion: Ghrelin may accelerate ICa-L inactivation and prolong the recovery time after inactivation. Ghrelin inhibits ICa-L in a dose-dependent manner.

BACKGROUND:The discovery and concept of pulp tissue-derived stem cells is beneficial to the understanding of tooth development and regeneration and repair mechanisms from the cellular level. OBJECTIVE:To understand the induced differentiation capacity and induced conditions in vitro of human dental pulp stem cells into neuron-like cells. METHODS:Pulp tissue was separated from human healthy third molars. Single cellsuspensions were prepared and seeded into 6-wel plates containing alpha-modified minimum essential medium supplemented with 15%fetal bovine serum. Subconfluent cultures (first passage) of colony forming cells were induced with butylhydroxy anisole, forskolin,β-mercaptoethanol, basic fibroblast growth factor. RESULTS AND CONCLUSION:Immunofluorescence and reverse transcription-PCR assay showed that human dental pulp stem cells positively expressed stro-1, Col-I, dentin sialoprotein after 2 weeks of induction. Nestin and neuron-specific enolase were strongly expressed, but the gingival fibroblasts were negatively expressed. It indicates that adult stem cells in human dental pulp have a high neuron-like celldifferentiation potential under a certain inductive condition.

Objective To study the risk factors of severe hand-foot-mouth disease (HFMD) among children.Methods The clinical data of 1 570 children with HFMD at Linyi People's Hospital in Shandong Province in 2011 were collected,retrospectively.The data were analyzed using univariate and multivariate Logistic regression.Results The mean age of severe HFMD (including severe and critical HFMD) was (25.0± 14.0) months old,predominantely aged between 1 and 5 years old,while mild HFMD was (27.1±15.8) months (t'=-2.717,P=0.007).There were 61.0％ and 65.9％ boys in two groups,respectively (x2 =3.894,P=0.048).Fever,convulsion,tremor,nausea and vomiting were more frequently seen in severe HFMD.The neutrophil count and the level of creatine kinase in severe HFMD were both significantly higher than that in mild HFMD.Univariate analysis revealed that age (odds ratio [OR]=1.799,95％CI:0.984-1.997),girl sex (OR=1.234,95％CI:1.001-1.522),high fever (OR=2.110,95％CI:1.816-2.452),convulsion (OR=1.878,95％CI:1.578-2.236),nausea and vomiting (OR=1.760,95％CI:1.456-2.128),neutrophil count (OR=1.031,95％CI:1.025-1.037) and creatine kinase (OR=1.002,95％CI:1.001-1.003) were risk factors for severe HFMD.Multivariate Logistic regression showed that high fever (OR =1.751,95％ CI:1.487-2.062),convulsion (OR=1.451,95％CI:1.204-1.749),nausea and vomiting (OR=1.269,95％CI:1.027-1.568),neutrophil count (OR=1.028,95％CI:1.021-1.035) were independent risk factors.Conclusions Body temperature,neurological manifestations and trend of neutrophil counts should be carefully monitored in children with HFMD.Prevention of the development of severe HFMD mainly relies on the identification of risk factors and adoption of precautions in time.

Objective To study the effects of Aralia chinesis on the proliferation and viability of fibroblasts and on the production of the hyaluronic acid(HA)in the cultured supernatant,and to explore its anti-hepatofibrosis mechanism.Methods NIH3T3 fibroblasts,which were cultured in vitro by routine method and were used as the substitutive model of hepatic stellate cells(HSC),were cultured with the rats serum containing Aralia chinese.The effects of Aralia chinesis serum on the cell proliferation was measured by methabenzthiazuron(MTT)assay and the HA content in cultured supernatant was detected by radioimmunoassay.Results Aralia chinesis serum showed no significant toxicity on NIH3T3 fibroblasts.5 %concentration serum of Aralia chinesis inhibited the cell proliferation and the synthesis of HA significantly(P

The inhibitory effects of diallyl sulfide (DAS) derived from allicin on in vitro and in vivo proliferation of human osteosarcoma MG-63 cells and the action mechanism, and the influence of DAS on invasive capability of MG-63 cells were investigated in order to search for the novel medicines for osteosarcoma. In the in vitro experiment, MG-63 cells were treated with different concentrations of DSA, and the morphological changes of MG-63 cells were observed under an inverted phase microscope. MTT method was used to assay the proliferation of MG-63 cells. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to detect the VEGF mRNA expression level in MG-63 cells. By using Transwell invasion assay, the influence of DAS on invasive ability of MG-63 cells was tested. In the in vivo experiment, the nude mice MG-63 cells tumor-bearing model was established, and different concentrations of DAS were injected beside the tumor. Twenty-one days after treatment, the mice were killed, the tumor size and tumor inhibition rate were calculated. The microvessel density (MVD) was determined by using immunohistochemistry. In the in vitro experiment, different concentrations of DAS could obviously inhibit proliferation of MG-63 cells in a time- and concentration-dependent manner. RT-PCR revealed that the expression levels of VEGF mRNA in DSA groups (different concentrations) were significant reduced as compared with those in control group (all P<0.05). Transwell invasion assay indicated that in 20 and 40 μg/mL DAS groups, the number of migratory cells was 91.4±8.3 and 81.8±7.4 respectively, which was significantly declined as compared with that in control group (150.4±14.7, both P<0.05). In the in vivo experiment, DAS could significantly suppress the growth of MG-63 tumor-bearing tissue. Immunohistochemistry demonstrated that different concentrations (20 and 40 μg/mL) of DAS could significantly decrease MVD of MG-63 tumor-bearing tissue (all P<0.05). It was suggested that DAS could inhibit the growth of MG-63 cells probably by suppressing the expression of VEGF mRNA.

Objective To study the morphological variations of maxillary sinus and to provide anatomical basis for maxillary posterior teeth implantation and extraction usingcone-beam computed tomography (CBCT) scan-ning. Methods CBCT images from 403 patients were reviewed. The minimal distance from sinus floor to alveolar ridge wasmeasured and the number of patients with clinical features such as having sinus septum , mucosal thicken-ing and posterior teeth roots extend into maxillary sinus were counted. Results The mean value of the minimal dis-tance from maxillary sinus floor to alveolar ridge was about 8.3 mm, there was no significant statistical difference between sex, but there was a significant statisticalsignificant difference between the left sides and the right sides. Occurrences of sinus septum, mucosal thickening and roots into sinus were 98.6%, 31% (in which 56.3% of mu-cosal thickening were related to apical periodontitis) and 47%. there was no significantstatisticaldifference between the two sides in males females. Conclusions The morphological variations of maxillary sinus are significant. Chron-ic inflammation in maxillary sinuis is closely related to the position of maxillary posterior teeth roots and periapical lesions.

Objective To investigate MVD, TSP-1, TGF-?1 expression in GH-Secreting Pituitary Adenomas by immunohistochemistry, and to correlate data with clinical characteristics.Methods The protein expression of TSP-1, TGF-?1 in 48 surgical specimens (21 invasive cases; 27 non-invasive cases) of pituitary adenomas was measured using immunohistochemical method. The relationship between the expression and clinical properties was examined. MVD was measured by detecting CD34.Results Compared with the noninvasive group, no difference of expression of CD34(t=2.257; P=0.083) was observed. The expression of TSP-1 in invasive group was low. The expression of TGF-?1 was higher in invasive cases than that in noninvasive ones. The expression of TGF-?1 had positive correlations with MVD; but there was no correlation between the expression of CD34 and the invasion of pituitary adenomas. In addition, MVD count was not associated with the expression of TSP-1. Size, sex or rate of recurrence did not influence MVD and TSP-1 expression. Conclusion MVD values do not necessarily represent angiogenesis in pituitary adenomas. TGF-?1 may increase MVD, and TSP-1 does not affect MVD in pituitary adenomas and angiogenesis may be regulated by other pathway.