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 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.

Objective To investigate effects of histone deacetylase 1 (HDAC1) gene silencing on proliferation and apoptosis of pancreatic cancer cell line PaTu8988. Methods The PaTu8988 cells were cultured and divided into control group (untreated), negative control group (treated with 30 nmol/L negative siRNA), low HDAC1 group (treated with 15 nmol/L HDAC1 siRNA) and high HDAC1 group (treated with 30 nmol/L HDAC1 siRNA). The real-time PCR and Western blot were used to detect the efficiency of HDAC1 gene silencing on mRNA and protein levels ,respectively, at 48 hours after transfection of HDAC1 siRNA. Cell proliferation and apoptosis were evaluated using cell counting kit and flow cytometry, respectively. Results Forty-eight hours after transfection of HDAC1 siRNA, the expression of HDAC1 mRNA level in PaTu8988 cells was 46.1%±6.1% in low HDAC1 group and 32.3%±1.4% in high HDACI group, which were lower than that in control group (100.0%±3.4%) and negative control group (87.4%±28.3%,P<0.05). The expression of HDAC1 protein was higher in control and negative control groups than in low and high HDAC1 groups. Cell survival rate was 100.0%±17.1% in control group, 87.1%±5.0% in negative control group, 68.7%±4.7% in low HDAC1 group and 61.6%±2.0% in high HDAC1 group with significant difference (P<0.05). Meanwhile, the cell apoptic rate in control (4.20%±0.95%) and negative control (4.59%±1.26%) groups was lower than that in low (10.09%±1.36%) and high (11.19%±6.07%) HDACI groups (P<0.05). Conclusions HDACI siRNA can effectively and specifically inhibit the expression of HDAC1 and proliferation of PaTu8988 cells and induce cell apoptosis.

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 clinic significance of HDAC1 expression in human pancreatic carcinoma.Methods 30 samples of pancreatic carcer tissues and paracancerous tissues were collected.HDAC1 expression was detected by real-time PCR and immunohistochemistry techniques.The relationships between clinicopathological findings and the expression of HDAC1 were analyzed. Results The RQ level of HDAC1 mRNA expression in human pancreatic carcinoma tissues and paracancerous tissues was 2.60(0.42～12.81)and 1.02(0.19～3.58),respectively(P=0.001).The percentage of positive cells expressed HDAC1 protein in human pancreatic carcinoma tissues and paracancerous tissues were(56 ±26)%and(6±6)%,respectively(P=0.000).The highly expressed HDAC1 correlated significantly with an advanced TNM stage and lymph node metastasis.Conclusions In pancreatic carcinoma tissues,highly expressed HDAC1may indicate the status of advanced TNM stage and poor prognosis.

Objective DNA methytransferase 1(DNMT1)is highly expressed in many cancers and lowly expressed in normal adult cells.This study was to assess effects of DNMT1 gene silencing on proliferation and apoptosis of pancreatic cancer cell line PaTu8988.Methods It is divided into four groups:Control group,Lipofectamine 2000 group,Negative control siRNA(N-siRNA)group(30 nM)and siRNA group(30nM).The expression levels of DNMT1 mRNA were detected by real-time PCR to assess the efficiency of DNMT1 gene silencing.Cell proliferation was analyzed by WST-8 assay;Cell apoptosis was evaluated by flow cytometry.Results Relative to Control group,48h after transfection of DNMT1 siRNA,The inhibitory rates of DNMT1 mRNA levels in PaTu8988 cells was(86.0±4.3)%.Cell survival rate was declined to 47.6±5.6%from Control group(100.7±3.0%),Lipofectamine 2000 group(64.5±6.8%),N-siRNA group(68.1±4.1%)(P＜0.05);Cell apoptosis rate was increased from Control group(7.51±1.12)%to siRNA group (14.94±2.89)%(P＜0.05),respectively,48h after transfection of DNMT1 siRNA.Conclusions DNMT1siRNA can efficiently and specifically knockdown the expression of DNMT1 mRNA and inhibit the proliferation of PaTu8988 cells,and induce cell apoptosis.It provides evidence for gene therapy of pancreatic cancer.