Objective To observe the effects of electroacupuncture at Zusanli (ST36) on lower limbs motor function in patients with stroke. Methods 63 patients were evenly randomized into electroacupuncture group and control group. All the patients received routine rehabilitation training and at the same time,patients in electroacupuncture group, received electroacupuncture at zusanli (ST36). They were assessed with Fugl-Meyer Assessment (lower limb, hip, knee and ankle), gait analysis (gait frequency, gait speed and step length of both sides) and lower limb Composite Spasm Scale (CSS) before and after treatment. Results The scores of Fugl-Meyer Measure, gait analysis, and CSS improved in both groups, and more significantly in electroacupuncture group. Conclusion Electroacupuncture at zusanli (ST36) can significantly improved lower limb motor function without worsening the spasm.

Objective To investigate the effects of trichostatin A (TSA) on cell proliferation and cell cycle in human gastric cancer cell line SGC-7901 in vitro and its mechanism. Methods SGC-7901 cells were treated with 0.1, 0.5 and 2.0 μmol/L of TSA for 24 hrs. Growth inhibition rates of cells were measured by MTT assay and cell cycles were detected by flow cytometery (FCM). Expressions of cyclin D1 and p21 mRNA were measured by real-time PCR. Results The proliferations of SGC-7901 cells were inhibited when treated with TSA for 24 hrs. The inhibition rates in groups treat with 0.1, 0.5 or 2.0 μmol/L of TSA were 3.52%±6.11%, 13.29%±4.13% or 14.24%±2.80% ,respectively. The cell percentage of G0/G1 phase were higher in 0. 5 pznol/L group (71.26%±0.51%) and 2.0 μmol/L group (71.03%±0.12%) compared with control group (51.12%±1.17%). The cell percentage of S phase were lower in 0.5 μmol/L group (13.55%±0.44%) and 2.0 μmol/L group (10.63%±0.63%) compared with control group (34.60%±0.60%). The expression of cyclin D1 mRNA was down-regulated, whereas p21 mRNA expression was up-regulated. Conclusions TSA inhibits SCG-7901 gastric cancer cell proliferation by affecting the cell cycle control gene eyclin D1 and p21 mRNA expressions, which induce G0/G1 cell phase cycle arrest and ultimately impact on the growth of tumor cells.

The purpose of education is to cultivate talents who can master the ability of self-learning lifelong. With the rapid development of multimedia technology, the knowledge carrier represented by micro curriculum plays a very important role in improving students' self-learning ability. In traditional Chinese medicine college, due to the short of time, weak learning foundation, the ability of self-learning is hard to improve in the modern medical courses such as biochemistry. This is not conducive to the cultivation of modern talents of Chinese medicine. In this paper, we chose the biochemistry teaching in TCM college as an example, and discuss how we can make the application of micro courses reasonably in the teaching process. This study was regarded as a starting to improve the students' self-learning ability effectively.

Objective To explore the effect of acupuncture on stroke patients in hemiplagic spasm period.Methods63 stroke patients in hemiplagic spasm period were randomly divided into the electro-acupuncture group(n=31) and control group(n=32).All patients of two groups received routine rehabilitation training,but those of the electro-acupuncture group were added with electro-acupuncture at "Zusanli"(ST36).The composite spasticity scale(CSS) score,H/Mmax and muscular compliance of two groups before and after treatment were observed and compared.ResultsThe CSS scores and H/Mmax and grading gastrocnemius muscular compliance of the electro-acupuncture group were superior to that of the control group(P<0.05～0.01).ConclusionAcupuncture can decrease muscular tension and increase motor function of stroke patients in hemiplagic spasm period.

ObjectiveTo assess the effects of DNA methyhransferase 1 ( DNMT1 ) gene silencing on DNMTs activity and methylated CpG sites of hMLH-1 in pancreatic cancer cell line PaTu8988.Methods DNMT1 siRNA and negative control siRNA was constructed by Ambion Company of United States.Then they were transfected into pancreatic cancer cell line PaTu8988 at the concentrations of 15,30 nmol/L,and the cells without transfection was used as the control group.Real-time PCR and Western blotting were applied to detect the DNMT1 mRNA and protein expression,and DNMTs activity was detected by using DNMTs activity assay kit.Change of methylation of CpG island of hMLH-1 was detected by bisulfite sequencing PCR (BSP).The expression of hMLH-1 mRNA was detected by Real-time PCR.ResultsAt 48 h after transfection,Realtime RT-PCR analysis showed that the levels of DNMT1 mRNA in DNMT1 siRNA group ( 15 nmol/L) and DNMT 1 siRNA group (30 nmol/L) were 0.573 ± 0.026 and 0.143 ± 0.044,which were significantly lower than those in control group 1.020 ±0.217 and negative siRNA 15 nmol/L group 0.900 ±0.475,and negative siRNA 30 nmol/L group 0.938 ± 0.327 (P ＜0.05 ).Western blotting analysis showed that the level of DNMT1 protein of DNMT1 siRNA group was also lower than those of negative siRNA and control groups.DNMT activity in DNMT1 siRNA15,30 nmol/L groups was 0.364 ± 0.124and 0.250 ± 0.072,which were significantly lower than those in control group 0.931 ± 0.065and negative siRNA group 0.665 ± 0.055 and 0.472 ± 0.040.DNMT activity was positively correlated with DNMT1 mRNA expression ( r =0.69,P ＜ 0.01 ).DNMT1 RNA interference decreased 8 methylated CpG sites of hMLH-1 to 1 site.Concluslons DNMT1siRNA can specifically inhibit the expression of DNMT1 gene of PaTu8988 and DNMT activity,and can decrease methylated CpG sites of hMLH-1 gene.

Objective To investigate the effects of histone deacetylase 1 (HDAC1) gene silencing on cell cycle of pancreatic cancer cell line PaTu8988 and possible mechanism.Methods The PaTu8988 cells were routinely cultured and divided into control group,negative control siRNA group (c-siRNA),HDAC1 siRNA 15 nmol/L group and HDAC1 siRNA 30nmol/L group.After Liposomes 2000 transfection for 48 h,the Western blotting was used to detect the efficiency of HDAC1 gene silencing on protein levels and the expression of p21 protein.Cell cycle was evaluated by using flow cytometry.Results Compared with that of control group,the expression of HDAC1 protein in PaTu8988 cell of HDAC1 siRNA 30nmol/L group was significantly decreased,and the expression of p21 protein was significantly increased; the percentage of G2/M phase cells were significantly decreased[(21.48 ±3.67)％ vs.(28.28 ±2.94) ％,P＜0.05]; while the percentage of S phase cells were significantly increased[( 50.20 ± 6.85 ) ％ vs.( 32.49 ± 2.78 ) ％,P ＜ 0.05].Conclusions HDAC1 siRNA can efficiently and specifically inhibit the expression of HDAC1 in PaTu8988 cells,and can induce S phase cells arrest,which may be related with up-regulation of p21 protein expression.

The mechanism of injury on the human glomerular endothelial cells (ciGENC) induced by preeclampsia serum was investigated. Concentration of maternal serum sFlt-1 protein was detected by ELISA. Fluorescently-labeled bovine serum albumin infiltrating through lower chamber of Transwell was measured by multifunction microplate reader. Morphologic change of ciGENC was observed under inverted phase contrast microscope. The concentration of sflt-1 in preeclampsia groups was significantly increased as compared with control group (P<0.01). Permeability in preeclampsia groups was significantly increased as compared with control group (P<0.01). By contrast with severe preeclampsia group, the permeability of ciGENC monolayer in mild preeclampsia group was decreased significantly (P<0.05). Intervention of exogenous VEGF significantly decreased permeability of ciGENC in preeclampsia groups. It was concluded that sFlt-1 increased ciGENC permeability by damaging integrity of endothelial barrier function.

This study investigated the effect of epigenetic modification of maspin on extravillous trophoblastic function. The mRNA expression of maspin in placentae from normotensive and preeclamptic pregnant women was detected by RT-PCR. TEV-1 cells, a human first-trimester extravillous trophoblast cell line, were cultured and treated with CoCl(2) (300 μmol/L) to induce chemical hypoxia and with 5-aza (500 nmol/L) to induce demethylation. The mRNA expression of maspin in TEV-1 cells subjected to different treatments was determined by RT-PCR, and the proliferative and migratory abilities of TEV-1 cells were assessed by cell counting kit-8 (CCK-8) and Transwell assays. Our results showed that the maspin mRNA expression level in placentae from preeclamptic women was much higher than that from normotensive women. CoCl(2) or 5-aza could up-regulate the mRNA expression of maspin and significantly suppress the proliferation and migration of TEV-1 cells. It was concluded that the epigenetic modification in promoter region of maspin contributes to incomplete trophoblast invasion, which offers a novel approach for predicting and treating placental dysfunction.

Objective To investigate the expressions of DNA methyltransferase 1 (DNMT1) in pancreatic carcinoma and its clinical significance.Methods 30 samples of pancreatic cancer tissues and paired para-cancerous tissues were collected from patients who underwent curative pancreatectomy.The levels of DNMT1 mRNA were detected by real-time RT-PCR.Expressions of DNMT1 protein were detected by streptavidin peroxidase immunohistochemistry.The relationships between expression of DNMT1 and clinicopathological findings were analyzed.Results The value of relative quantification (RQ) of DNMT1 mRNA in human pancreatic cancer tissues was 2.32 (1.17 ～ 5.17 ), which was significant higher than 0.78 (0.07 ～3.14) in para-cancerous tissues(P ＜0.05).The index of expression of DNMT1 protein in human pancreatic cancer tissues was (54.5 ±21.2)% ,which was significant higher than( 10.9 ± 15.0)% in paracancerous tissues (P ＜ 0.01 ).Patients were divided into the high DNMT1 group (n = 19) with above 54.5% of the DNMT1 positively cancer cells and the low DNMT1 group ( n = 11 ) with less than 54.5% of the DNMT1 positively cancer cells.The high expression of DNMT1 was correlated significantly with clinical staging ( x2 =6.897, P = 0.029), lymph node metastasis ( x2 = 4.739, P = 0.029) and neural invasion ( x2 = 5.44, P =0.020).On the other hand, no association between DNMT1 expression and age, gender, tumor location,tumor size, tumor differentiation, the serum CEA and CA19-9 levels could be found.Conclusions DNMT1 mRNA and protein was highly expressed in pancreatic cancer tissues.High expression of DNMT1 might be related to the aggressiveness of pancreatic cancer, lymph node metastasis and neural invasion.

Objective To investigate the effect of trichostatin A (TSA) on inhibition of cell proliferation and cell cycle of human pancreatic cancer cell line PaTu-8988 in vitro. Methods PaTu-8988 cells were treated with different concentration TSA (0.1,0.5,2.0μmol/L), and blank control group was used. The growth inhibition rates of cells were measured by WST-8 method. Cell cycle and apoptosis were detected by flow cytometry; the expressions of p21 mRNA and HDAC1 mRNA were measured by Real-Time PCR. Results Survival rates of TSA 0.1μmol/L,0.5μmol/Land2.0μmol/L group was (88.5±4.2)%, (79.7±5.O)% and (64.3±7.2)%, respectively, which were all significantly lower than (100.0±4.2)% of the blank control group (P<0.01). Cells in the group of TSA O.1μmol/L and 0.5μmol/L were hindered at G1 phase, while (50.29±7.53)% of the cells in the group of TSA 2.0μmol/L were hindered at G2/M phase. The expression rates of p21 mRNA in different TSA group were 5.29±1.16, 7.79±0.41, 8.61± 0.73, respectively, which were higher than that in the control group (1.00±0.08, P<0.01); the expression rates of Cyclin DI mRNA were 1.13±0.12, 0.42±0.06, 0.19±0.06, respectively, which were higher than that in the control group (1.00±0.07, P<0.05). Conclusions TSA may up-regulate the expression of p21 and down-regulate the expression of cyclin D1, and induce cell cycle blockade.