Objective To observe the effects of intensive training at different intensities on the expressions of semaphorin 3A ( Sema 3A) and its receptor neuropilin ( NP-1 ) and the cell apoptosis in cerebrum after cerebral ischemia-reperfusion in rats,and to investigate the possible mechanism of intensive training in recovery of motor function after cerebral ischemia-reperfusion in rats.Methods To establish animal model of cerebral ischemia-reperfusion in rats,the intraluminal thread method was applied to cause left middle cerebral artery occlusion (MCAO) for 2 h and before reperfusion.After cerebral ischemia-reperfusion model were established for 24 h,60 male model Wistar rats were randomly divided into training group 1 ( swimming for 5 min once a day),training group 2 ( swimming for 10 min once a day),training group 3 (swimming for 10 min twice a day) and control group (no training) ; another 15 rats assigned to the sham-operation group were subject to no MCAO and no training.Neurological function was evaluated by Garcia scores,and terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) was performed to dectect the cortical cell apoptosis.Expressions of neural growth inhibition factor Sema 3A and its receptor NP-1 were detected by immunohistochemistry.Results The neurological function in sham-operation group was normal.The differences of Garcia scores at different time points beween sham-operation group and control group were significant (P ＜ 0.01 ).Garcia scores in all training groups,were significantly higher than those in controls at the 7th and 14th d after swimming training ( P ＜ 0.01 ),especially in training group 3 the Garcia scores were ( 12.80 ± 0.45 ),( 15.20 ± 0.45 ),( 16.80 ± 0.45 ),respectively,at the 3rd,7th and 14th d after swimming training.The rates of positive cell of Sema 3A,NP-1 and TUNEL indexes in all training groups were lower than those in controls at the 3rd,7th and 14th d after swimming training (P ＜ 0.01 ),especially in the training group 3.At the 3rd,7th and 14th d after swimming training in training group 3,the rates of TUNEL indexes positive apoptosis cells were ( 29.43 ± 1.38 ) ％,( 22.30 ± 1.21 ) ％,( 17.58 ± 1.70) ％,respectively,the positive cell rates of Sema 3A were ( 19.64 ± 1.17) ％,(9.73 ± 3.83)％,(8.24 ± 0.87)％,respectively,the positive cell rates of NP-1 were ( 33.95 ± 6.86) ％,( 27.95 ± 1.29 ) ％,( 18.90 ± 1.44 ) ％,respectively,the reduction of positive cells expressions in training group 3 was significantly more obvious compared with other training groups (P ＜ 0.01 or 0.05).Conclusions Rehabilitation training can reduce the expression of positive cell of Sema 3A,NP-1 and TUNEL indexes in rats after cerebral ischemia-reperfusion and can improve motor function recovery and facilitate neural plasticity.The more intensive the training,the better the effects.

OBJECTIVETo establish the quality standard of Entadae Semen, and provide scientific basis for its quality control.

METHODTLC and HPLC were used for qualitative identification and quantitative analysis of phaseoloidin and entadamide A-O-β-D-glucopyranoside in Entadae Semen. The test of water content, ash and ethanol-soluble extractives of Entadae Semen was carried out according to the methods recorded in appendix of Chinese Pharmacopeia (2010 edition).

RESULTThe TLC was well separated with clear spots. The linear range of phaseoloidin was between 0.014 to 2.747 g x L(-1) (r = 0.999 6, n = 9) with an average recovery rate of 101.06% (RSD 0.90%, n = 6); the linear range of entadamide A-O-β-D-glucopyranoside was between 0.002 to 0.452 g x L(-1) (r = 0.999 7, n = 9) withan average recovery rate of 101.52% (RSD 1.09%, n = 6). The content of phaseoloidin in sample is between 5.12% to 9.24%, entadamide A-O-β-D-glucopyranoside is between 0.55% to 2.17%, alcohol-soluble extracts is between 30.9% to 45.2%, water is between 6.6% to 8.6%, and total ash is between 2.4% to 2.9%.

CONCLUSIONThe established standard is acceptable for quality control of Entadae Semen.

China ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Fabaceae ; chemistry ; Quality Control

BACKGROUND:Hyperbaric oxygen(HBO)treatment promotes the proliferation and differentiation of endogenous neural stem cells in neonatal rats following hypoxic/ischemic brain damage(HIBD).The Wnt signaling pathway is associated with neurogenesis.However,there are few data recording the role of HBO in the differentiation of neural stem cells in vitro.OBJECTIVE:To observe the effect of HBO on differentiation and Wnt3 expression of bone marrow mesenchymal stem cells(BMSCs).METHODS:BMSCs were isoiated and cultured.The rat BMSCs of passages 3-5 were cultured in DMEM/F12(1:1)medium with basic fibroblast growth factor,epidermal growth factor and B27 for 24 hours.The induced BMSCs were randomly divided into two groups:control group(no treatment)and HBO group(HBO,0.10 MPa,60 minutes stabilizing pressure with at least 90% oxygen).The neuron specific encloase(NSE),glial fibrillary acidic protein(GFAP)and 04 marked oligodendrocyte immunocytochemistry were detected by immunofluorescent staining,and Wnt3 protein expression was detected by Western-blot.RESULTS AND CONCLUSION:BMSCs cultured in classic medium of neural stem cells could significantly induce the expression of nestin.The expression of NSE and 04 of HBO group was greater than control group(P＜0.01),but GFAP expression displayed no significant difference between the groups(P＞0.05).Western blot showed HBO could enhance the Writ3 expression (P＜0.05).Results show that HBO can induce BMSCs to differentiate into neural cells and oligodendrocyte,which is correlated with the activation of the Wnt3 protein.

Objective To investigate the current status about nursing undergraduate students' knowledge,attitude and practice (KAP)regarding nosocomial infection.Methods The self-administered questionnaires were employed to survey 108 undergraduate nursing students on the basis of a simple random sampling method.Results In the knowledge dimension,the nursing students earned 78.3％ of accuracy rate when responding to the questionnaires.The students also demonstrated positive attitudes towards nosocomial infection and occupational safety,particularly the female students.In terms of practice,the students performed relatively poor as 36.1％ of the students were unclear about the classification of medical garbage and 22.2％ of the students used their non-clean hands to touch their eye-glasses.Conclusions The undergraduate nursing students have demonstrated adequate knowledge and proper attitude towards nosocomial infection and occupational safety,however,some behaviors need to be changed.Nursing schools and hospitals should be aware of these findings and provide more training programs regarding nosocomial infection and occupational safety so that they can help students formulate good habits to prevent and control nosocomial infection.

Objective:To prepare poly-L-lactic acid(PLLA)electrospun nanofibers carrying icariin(ICA)(ICA /PLLA)and to evaluate the effects of the ICA /PLLA on MC3T3-E1 cells.Methods:ICA solution was dispersed into PLLA solution,and electrospun fibers were fabricated by W/O emulsion method.The morphology of ICA /PLLA was observed by SEM.The in vitro release kinetics of ICA /PLLA was examined.The attachment of MC3T3-E1 cells on ICA /PLLA was examined by propidiumiodide(PI)labling and ob-served under fluorescent microscope.The proliferation of the cells was measured by MTT assay.The differentiation of the cells was ob-served by alkaline phosphatase (ALP)assay.Results:In vitro,ICA was effectively released from ICA /PLLA for 22 days,cells were attached well on the surface in all groups,ICA did not affect the proliferation of MC3T3-E1 cells(P >0.05),but increased the ALP activity(P <0.05)of the cells.Conclusion:ICA /PLLA can effectively control the release of ICA and promote the differentiation of MC3T3-E1 cells.

Objective To investigate the expression of Fascin in early-stage NSCLC, evaluate the relevance between Fascin expression level and prognosis.Methods The immunohistochemistry method was used to assess the expression of Fas-cin in 111 lung cancer FFPE tissues with stage Ⅰ and Ⅱ NSCLC.The relationship between Fascin expression and the clinico-pathological characteristics was analyzed.The prognostic significance of Fascin expression was evaluated with Kaplan-Meier sur-vival analysis.Results In the early-stage of NSCLC, the positive rate of Fascin expression was 64.8%, no expression in the paracarcinoma tissue.The positive rate of squamous cell carcinoma was 78.7% and was significantly higher than that in adeno-carcinoma 48.0%(P<0.01).Whether in squamous cell carcinoma or adenocarcinoma group, the expression of Fascin was correlated significantly with lymph node metastasis tumor stages and DFS(P<0.05).And the positive expression of Fascin was an independent risk factor of poor prognosis for patient with NSCLC .Conclusion Fascin is expected to be a biomarker for the prognosis of patients with early-stage NSCLC.

Objective To investigate the expression of corticotropin-releasing factor (CRF) and its receptors including CRFR1 and CRFR2 on mouse mesenteric lymph nodes dendritic cells (MLNDC), and to analyze their effects on the biological phenotypes of intestinal dendritic cells .Methods The MLNDCs were isolated from C57BL/6 mice by using magnetic bead sorting .The purity of CD11c+DCs was identified by flow cytometry .The double-labeling immunofluorescence and the reverse transcriptase-polymerase chain reaction (RT-PCR) were performed to detect the expression of CRF , CRFR1 and CRFR2 on MLNDCs.The MLNDCs were exposed to CRF with or without the interference of CRFR 1 and CRFR2 antagonists .Flow cy-tometry was used to measure the changes of surface molecules ( MHCⅠ and MHCⅡ) and co-stimulatory molecules (CD80 and CD86).Results The CD11c+DCs accounted for (80.12±6.34)% of the isolate cells with a high cell viability of more than 90%.The expression of CRF , CRFR1 and CRFR2 at mRNA lev-el were detected in MLNDCs by RT-PCR.Results of the immunofluorescent staining assay indicated that both CRFR1 and CRFR2 were expressed on the surface of MLNDCs .The expression of CD86 on MLNDCs was inhibited by the treatment of MLNDCs with CRFR 1 antagonist , but enhanced by the treatment with CRFR2 antagonist .Conclusion Both CRF and CRFRs were detected in the MLNDCs isolated from the C57BL/6 mice.The CRF could alter the biological phenotypes of MLNDCs through binding to different CRFRs (CRFR1 and CRFR2), which affected the phenotypes of MLNDCs in opposite ways .

Objective: To investigate the expressions of programmed death ligand 1（PD-L1）in triple-negative breast cancer (TNBC) and its correlation with angiogenesis. Methods: 120 cases of TNBC patients who underwent surgery in the Fourth Hospital of Hebei Medical University from March 1, 2011 to June 1, 2012 were collected. The tumor tissues of patients were surgically resected and confirmed by pathology. PD-L1 protein expression in TNBC tissues of 120 patients was detected by tissue microarray combined with immunohistochemistry, and its relationship with various clinical indicators was analyzed. Blood vessels and lymphatic vessels were labeled withCD34andD2-40todetectmicrovesseldensity(MVD)andlymphaticvesseldensity(LVD)inTNBC.Results:Thepositiveexpression rate of PD-L1 in the tumor cells and interstitial infiltrating lymphocytes fromTNBC was 56.7% (68/120); No correlation was found between PD-L1 protein expression and the gender, age, histological grade, clinical stage, or tumor size of patients with TNBC (P>0.05), but related to the lymph node metastasis (P<0.05) and vascular thrombus (P<0.05). TNBC with high PD-L1 expression exhibited high incidence of lymph node metastasis and formation of vascular thrombus, and the expression of PD-L1 was positively correlated with MVD (r=0.500, P=0.02) as well as LVD (r=0.662, P=0.01). Log-Rank test showed that the survival time of TNBC patients with positive PD-L1 protein expression was significantly shorter than that of patients with negative expression (P<0.05). Cox multivariate analysis suggested that PD-L1 protein expression could be an independent prognostic factor for TNBC overall survival. Conclusion: PD-L1 plays an important role in TNBC angiogenesis and lymphangiogenesis, and is closely related to TNBC invasion and metastasis; blocking PD1/PD-L1 signal pathway is expected to be an effective new strategy for TNBC treatment.

Aim To explore new ways for developing anticancer drugs by the separation of pigment from Fu-sarium species JN158 ( Fusarium sp JN158 ) , the iden-tification of its structure, the screening of anticancer components and the study of its partial mechanism. Methods Pigment separation was done by HPLC, structural analysis by UV, IR, NMR, the screening of anticancer activity by MTT. Western blot was used to analyze the protein expression of CyclinD1, NF-κB, VEGF in tumor cells. Results The results showed that the pigment from Fusarium produced a total of six different peaks, of which peak Ⅵ was the anthocya-nins. Its molecular weight is about 382, molecular for-mula is C17 H18 O10 . According to investigation, this pig-ment was probably a new compound, which could in-hibit the proliferation of MCF-7 cells markedly ( IC50:0.011mmol·L-1 ,P<0.05;the control medicine ube-nimex IC50:10 mmol · L-1 ) in a concentration-de-pendent manner, and had no effect on human umbilical cord intravenous endotheliocyte ( HUVEC ) . The influ-ence on the gene expression of CyclinD1, NF-κB, VEGF in MCF-7 cells varied with the concentration of this compound. The Western blot results showed that VI pigment compound inhibited CyclinD1, NF-κB, VEGF gene expression (P<0.05 or 0. 01),compared with the control group. Conclusion The Ⅵ pigment compound from Fusarium sp JN158 could inhibit MCF-7 proliferation by inhibiting CyclinD1, NF-κB, VEGF gene expression. The compound may be a promising compound against breast cancer.

Objective To investigate the biomechanical and pathological changes in vivo in the taut bands (TB) of biceps femoris in rats after repeated low-frequency electrical stimulation.Methods Twenty-eight Wistar rats were randomly divided into a control group,an electrical intensity-dependent fatigue group,which were subject to electric intensity-dependent fatigue test,and an electrical frequency-dependent fatigue group,which were subject to electrical frequency-dependent fatigue test.After fatigue tests,the taut band of the biceps femoris and the non-taut band of the contralateral biceps femoris were harvested for pathological observation.The maximum contraction force (MCF),electrical intensity-and frequency-dependent fatigue characteristics and any pathological changes in the TBs were assessed and compared to the non-taut band region of the other biceps femoris.Results The MCF at the 15th and 20th stimulation (1.42 ± 0.28 g and 0.93 ± 0.54 g respectively) were significantly lower than that at the 1 st and 5th stimulation of the TBs.High stimulation intensity (HSI) at the 15th and 20th stimulation (3.76 ± 0.71 V and 3.44 ± 0.97 V) were also significantly lower than at the 1st TB stimulation.At the 10th,15th and 20th stimulation of the TBs,MCF and HSI were both significantly lower than in the bands which were not tight.In the frequency-dependent fatigue stimulation tests,the frequency which generated the MCF of the TBs was significantly lower than in the bands which were not tight,while the MCF of the TBs was significantly higher than that of non-TBs.After either intensity or frequency fatigue testing,more severe edema,uneven cytoplasmic death and degeneration of muscle fibers were observed in sections from TBs than from the bands which were not tight.Conclusions Taut muscle bands are significantly less fatigue-resistant than normal muscle fibers.Taut bands may contribute to the fatigue of myofascial pain syndromes.