Objective:To establish the fingerprint of the intermediate of Dalitong Granules; To control its quality with QAMS method.Methods:Gradient elution was performed by HPLC on Wondasil C18 superb column (4.6 mm × 250 mm, 5 μm). The mobile phase was 0.02% acetonitrile - acetic acid aqueous solution, gradient elution. The detection wavelength was 325 nm and the injection volume was 10 μl. The column temperature was 30 ℃, and the flow rate was 1.0 ml/min. Chemical recognition patterns were combined for analysis, and QAMS method was used for content determination.Results:Through methodological investigation, the fingerprint of Dalitong Granule Extract had good repeatability. The 10 batches of Dalitong Granule Extract could be divided into 3 categories through clustering analysis and principal component analysis. The relative correction factors of four components in Dalitong Granule Extract were established. Compared with external standard method, the established QAMS method has higher reliability.Conclusion:The established fingerprint and QAMS method can be used for the quality control of Dalitong Granule Extract.

Objective:To detect and verifica the gene profile difference of microRNA-146a (miR-146a) and its role in the pro-liferation of vascular smooth muscle cells (VSMCs) by gene chip technology. Methods: Artificially synthesized miR-146a mimics(50 nmol/L) ,miR-146 inhibitor ( 50 nmol/L ) , scramble ( 50 nmol/L ) and PBS were transfected into cultured primary rat VSMCs in vitro. After transfection,Real time PCR was used to measure the levels of miR-146a and the cell counting kit 8(CCK8) was employed to investigate the proliferation of VSMCs. The VSMCs interfered by miR-146a inhibitor or miR-146a control were examined by gene chips and the profile of gene were analyzed by bioinformatics technology to detect the different genes and signal transduction pathway. The changes in mRNAs and proteins were accessed separately by Real time PCR and Western blot. Results: Compared with sham and control VSMCs,miR-146a expression level was significantly decreased in treatment with miR-146a inhibitor(P<0. 01),as well as optical density(OD) was also shown remarkably down regulated simultaneously(P<0. 05). The investigation of gene profile revealed that the p53 signal pathway was up-regulated in VSMCs interfered by miR-146a. The mRNA and protein expression levels of p53, caspase3 and PTEN in p53 signal transduction pathway didn′t show significant differences(P>0. 05),however,the mRNA and protein expression levels of cyclin D1 significantly increased in treatment with miR-146a mimics VSMCs group and decreased in miR-146a inhibitor VSMCs group ( compared with sham VSMCs group, both P<0. 05 ) . Conclusion: Our data indicated that miR-146a may promote the proliferation of rat VSMCs by up-regulating cyclin D1 expression.

BACKGROUND:Previous studies have shown that a certain dose of acidic fibroblast growth factor can promote skeletal muscle satelite cel proliferationin vitro. OBJECTIVE:To investigate the effects of transfection with acidic fibroblast growth factor by electroporation on growth, proliferation and differentiation of skeletal muscle satelite cels. METHODS: Skeletal muscle satelite cels were cultured and purified, and then transfected with plasmid pSectag-GFP-aFGF by electroporation. The expression of green fluorescent protein was observed under fluorescence microscope, and the transfection efficiency was calculated. After transfection, cel cycle was analyzed by flow cytometry to draw the growth curve of skeletal muscle satelite cels. Western blot assay was employed to measure protein level of acidic fibroblast growth factor. RESULTS AND CONCLUSION: (1) Immunocytochemistry detection: The skeletal muscle satelite cels were positive for a-sarcomeric actin. (2) Transfection efficiency: At 12 hours after transfection with pSectag-aFGF, several cels showed green fluorescence, and the green fluorescent expression reached the peak at 72-96 hours after transfection with a positive rate of about 90%. (3) Cel cycle: After electrotransfection, the proportion of cels at S phase in the electroporation group was higher than that in the control group (P < 0.05). (4) Cel growth curve: At 3 days after electrotransfection, the cels entered logarithmic growth phase but the proliferation slowed down at 5 days. (5) Differentiation capacity: There were fewer myotubes and aging cels in the electroporation group than the control group. (6) Western blot assay: Acidic fibroblast growth factor protein was highly expressed in the cels transfected with target gene detected by western blot assay. These findings indicate that by using electroporation method, acidic fibroblast growth factor can be transferred into skeletal muscle satelite cels and have a high-efficiency and long-term expression, which can promote the proliferation of skeletal muscle satelite cels and inhibit formation of myotubes.

BACKGROUND:Skeletal muscle satelite cels are muscle-derived stem cels with proliferation and differentiation potential distributing between the muscle cel membrane and the base film. Studies have shown that skeletal muscle satelite cels are of efficacy and safety, but the survival rate of the transplanted stem cels is very low, which greatly limits the application of skeletal muscle satelite cels. OBJECTIVE: To observe the effects of Fasudil on apoptosis of skeletal muscle satelite cels induced by H2O2. METHODS: Skeletal muscle satelite cels cultured in vitro were randomly divided into three groups including H2O2group, H2O2+Fasudil group (Fasudil group) and control group. Apoptosis rates were observed by flow cytometry. The concentrations of interleukin-4 and tumor necrosis factor-a in each group were detected by ELISA. Western blot was employed to measure the protein level of Bax in each group. RESULTS AND CONCLUSION: Compared with the H2O2group, a significant decrease was found in the apoptosis rate of cels, protein level of Bax, and concentrations of interleukin-4 and tumor necrosis factor-a in the Fasudil group (alP < 0.05). These findings indicate that Fasudil can play anti-apoptosis protection by inhibiting Rho-kinase signaling pathway, which may be related to the reduced expression of Bax.

AIM:To investigate the proliferation property of stable chemerin gene knockdown vascular smooth muscle cells ( VSMCs) and to explore its mechanism .METHODS:The normal VSMCs , chemerin gene interfering control VSMCs and stable chemerin gene knockdown VSMCs were divided into normal group , PDGF group, control group and knockdown group .The VSMCs in PDGF group were given platelet-derived growth factor-BB ( PDGF-BB) to initiate proli-feration.The cell counting and BrdU assay were employed to investigate the proliferation property of VSMCs .The mitogen-activated protein kinase ( MAPK) signal pathway was determined by Western blot .RESULTS:The cell number and BrdU A value in PDGF-BB-treated VSMCs significantly increased as compared with the normal VSMCs ( P<0.05 ) .Compared with the normal VSMCs , the chemerin knockdown VSMCs showed obviously decreased cell number and BrdU A value ( P<0.05).Simultaneously, no significant difference in the proliferation of VSMCs between the normal VSMCs and the control VSMCs was observed.No significant difference of the protein levels of p-ERK1/2, ERK1/2, p-p38 MAPK and p38 MAPK among 4 kinds of VSMCs was found .The protein level of p-JNK in PDGF-BB-treated VSMCs was up-regulated, while it was down-regulated in chemerin knockdown VSMCs compared with the normal VSMCs .CONCLUSION: Chemerin pro-motes the proliferation of mouse vascular smooth muscle cells by up-regulating p-JNK production .

Objective To evaluate the relationship between emergence agitation (EA) during recovery from general anesthesia and postoperative cognitive dysfunction (POCD).Methods Two hundred and eighty ASA Ⅰ or Ⅱ patients,aged 18-70 yr,weighing 52-80 kg,undergoing elective surgery,were included.Anesthesia was induced with midazolam,fentanyl,propofol and cisatracurium.The patients were tracheal intubated and mechanically ventilated.Anesthesia was maintained with remifentanil,propofol and cisatracurium.EA was assessed at 15-40 min after extubation by using Post-operative Quality Recovery Scale and the cognitive function was assessed at day 1 before operation and days 1-7 after operation.Patients were divided into POCD or nonPOCD group according to the occurrence of POCD.The general data of patients,preoperative complications and types of surgery were recorded.If there was significant difference between the 2 groups,the factor was analyzed using multi-factor logistic regression to select the risk factor for incidence of POCD.Results The incidence of POCD was 40.7 ％.The results of logistic regression analysis showed that the dangerous degree of the risk factors for POCD in order from high to low were emergence agitation,duration of anesthesia and age.Conclusion EA during recovery from general anesthesia is an independent risk factor for POCD.

objective To describe the clinical features of patients with posterior circulation ischemic stroke.Methods 216 patients with posterior circulation ischemic stroke admitted in our department during 2004-2006 were analyzed retrospectively.All patients were undertaken MRI on admission and responsble lesions were identified at the posterior circulation territories.The patients'clinical symptoms and signs were evaluated and the relationships between lesion locations and clinical characteristics were analyzed.Results The common symptoms of posterior circulation ischemic stroke were unilateral limb Weakness(81.9%),speech difficulty(46.3%),dizziness(33.8%),and unilateral limb numbness (31.O%).The common signs of posterior circulation ischemic stroke were unilateral limb weakness (81.9%),central facial or lingual palsy(61.1%),dysarthria(46.3%),unilateral limb sensory loss (31.0%),and ataxia(30.1%).The incidence of crossed paralysis was low(2.8%).Isolated vertigo was rare (1.4%).Predominant clinical features such as bulbar paralysis,unconsciousness,visual disorder and amnesia can help to localize the lesions.Typical brainstem syndromes had topographic meanings.Conclusions The clinical features of patients with posterior circulation ischemic stroke were complex.Predominant symptoms can help to diagnose the posterior circulation ischemic stroke.

Objective To investigate the major risk factors of posterior circulation infarction.Methods Clinical data from 216 patients with posterior circulation infarction were analyzed retrospectively. The patients were divided into follow groups (proxima1,middle, dista1 and combination group,or single , multiple, or unilateral, bilateral, or lacune infarct, non lacune infarct ) according to the infarcts locations on MRI.The risk factors in each group were analysed. Also,the major risk factors were compared with that from patients with anterior circulation infarction. Results In 216 patients with posterior circulation infarction, hypertension was the most common risk factor (76.9%), followed by diabetes mellitus (36.6% ),hyperlipedemia (30.1%), previous stroke history(26%), and heart disease(22.2%). The most common location of infarcts was distal territory (49%),followed by middle(24.5%) ,proxima1(6%). The average age of proximal group [(57.92?12.81) years] was significant lower than that of other groups(P