Aim To investigate the inhibition mechanisms of baclofen, a specific GABA B receptor agonist, on quantal glutamate release in the rat spinal dorsal horn neurons.Methods Whole-cell voltage-clamp technique was performed on dorsal horn neurons in rat spinal cord slice to record glutamatergic spontaneous miniature excitatory postsynaptic currents (mEPSCs). Baclofen action on quantal glutamate release was assessed by analyzing the change of mEPESC to baclofen perfusion.Results Baclofen(10 ?mol?L -1,50 s) depressed the frequency, but not amplitude distribution of glutamatergic mEPSCs, indicating baclofen presynaptic depression on glutamate release. The depression on frequency of mEPSCs persisted in Ca 2+-free solution, or in the presence of K + conductance blocker, 4-AP. On the other hand, the depression was occluded by forskolin, an activator of adenylate cyclase, but not protein kinase C (PKC) activator phorbol 12,13-dibutyrate (PDBu). N-ethylmaleimide (NEM), a sulphydryl alkylating agent, which destroys G protein, abolished baclofen depression.Conclusion Not presynaptic K +, Ca 2+ conductance or PKC, but G protein and/or cAMP pathway are involved in the baclofen depression on glutamate release in rat spinal dorsal horn;this depression might contribute to the analgesic action of baclofen at spinal level.

Objective The liver is the main target of colon cancer metastasis, and liver metastasis is important and difficult in colon cancer therapy.This study was to investigate the synergistic effects of Honghua injection combined with oxaliplatin in treatment of liver metastasis from colon cancer. Methods Spleen-preserving method was used to establish the model of liver metastasis from colon cancer.The model mice were randomly divided into 5 groups:control group, oxaliplatin group, low dose ( low dose of Honghua injection combined with oxaliplatin) group, medium dose ( medium dose of Honghua injection combined with oxaliplatin) group, high dose (high dose of Honghua injection combined with oxaliplatin) group (n=10).Oxaliplatin were given twice a week at a dose of 5 mg/kg.Honghua injection was given at doses of 2.5, 1.25 and 0.625 g/kg for consecutive 14 days, and the control group received sa-line.After 14 d, all the mice were killed, and the tumor inhibiting rates and liver metastasis rates were calculated.The levels of serum vascular endothelial growth factor ( VEGF) and hepatocyte growth factor ( HGF) were measured by ELISA, and the mRNA expressions of VEGF and HGF in liver were determined by PCR. Results The inhibition rate of low, middle and high dose group achieved 63.2%, 69.2%and 71.4%respectively, which was significantly higher than that of control group (P<0.01).Compared with oxaliplatin group,middle and high dose groups showed a significant higher inhibition rate (P<0.05).Compared with control group, the liver metastasis rates in middle and high dose groups significantly decreased from 90%to 60%and 50%respectively (P<0.05).The serum VEGF and HGF levels showed significant decrease in all treatment groups([83.4 ±25.6],[81.5 ±23.7],[75.8 ±19.4],[72.7 ±20.5] pg/mL),([56.2 ±13.8],[55.4 ±15.7],[48.9 ±13.6],[42.3 ±11.8]pg/mL) compared with control group ([125.3 ±36.2], [98.4 ±26.1]pg/mL) (P<0.01).The mRNA expressions of VEGF and HGF in liver were significantly decreased in all the Hong-hua injection combined with oxaliplatin groups compared with control group ( P<0.05) .Compared with oxaliplatin group, the serum VEGF level in high dose group showed a significant reduced mRNA expression of VEGF (P<0.05). Conclusion Honghua injec-tion can significantly enhance the inhibitory effects of Oxaliplatin on liver metastasis from colon cancer, and the mechanism may con-tribute to the inhibition of VEGF expression and the prevention of angiogenesis in liver.

Objective To construct the prokaryontic expression vector of the gene fragment which encodes the transpeptidase domain of penicillin binding protein 2a(PBP2a) of methicillin‐resistant Staphylococcus aureus(MRSA) ,and to express ,purify and i‐dentify the objective protein .Methods Strains of MRSA were isolated and identified from clinical samples ,according to the se‐quence of mecA gene recorded in GenBank ,the primers of mecA fragment which encoded the transpeptidase domain of PBP2a was designed .The gene fragment from MRSA was amplified by using polymerase chain reaction(PCR) and cloned into pET28a(+ ) plasmid .After being identified by enzyme digestion and sequencing ,the recombinant plasmid was transformed into the strain of Escherichia coli BL21(DE3)plysS .The expression of transpeptidase domain of PBP2a was induced by 0 .7 mmol/L IPTG ,the ex‐pressed products were purified by using Ni afinity chromatography ,then were analyzed by using Western blot .Results The recom‐binant expression vector was digested by BamHⅠ and EcoRⅠ ,and the products were at the expected size .The result of sequencing showed two bases undergoing mutation ,while there were no frameshift mutations .The expressed protein was identified by using SDS‐PAGE and Western blot ,a new protein band was visible at the relative molecular mass of 38 × 103 .Conclusion The corre‐sponding prokaryotic expression vector is successfully constructed ,and the transpeptidase domain of PBP2a is successfully ex‐pressed and purified .

Objective To screen the microsatellites with low occurrence rate of stutter band and establish the effective bovine STR typing system.Methods The tetranucleotide STR loci in bovine genome were searched with Tandem Repeat Finder software.Primers were designed and used to amplify these candidate loci and the PCR products were separated with electrophoresis.DNA samples from 100 head of unrelated cattle were typed.Results Among these candidate loci,6 bovine tetranucleotide STR loci showed high polymorphism,and their CDP and CPE value were 0.99995 and 0.859591 respectively.Conclusion The 6 bovine tetranucleotide STR loci can be used for bovine identification and parentage testing.

Objective: To develop a questionnaire that can well measure the concept of work-family enrichment. Methods: 225 valid data from 3 enterprises were collected in order to develop work-family questionnaire by exploratory factor analysis. 268 samples from other 10 enterprises were collected to test the validity and reliability of the questionnaire. Results: ① Work -family enrichment questionnaire was composed of two second -order factors: work to family enrichment and family to work enrichment. Each second -order factor was composed of two first -order factors: instrumental enrichment and psychological enrichment. ②The structure validity and criteria-related validity of this work- family enrichment questionnaire were good. ③ The Cronbach ? of work to family enrichment and family to work enrichment were 0.86、0.84. The Cronbach ? of the whole questionnaire was 0.89. Conclusion: Work-family enrichment questionnaire has good construct validity、criteria-related validity and internal consistency reliability.

Objective To investigate the effect of propofol on the high-voltage-activated calcium currents [ICa(HVA)] in rat hippocampal neurons. Methods Hippocampal neurons were prepared from Wistar rats and cultured. ICa(HVA) was recorded using whole-cell patch clamp technique. Different concentrations of propofol were added to the culture. The effect of propofol on ICa(HVA) Was evaluated. Results ICa(HVA) was inhibited by propofol in 300 μmol/L reduced peak ICa(HVA) by (24±6)%, (33 ±5) %, (36±7)% and(38±3)% respectively with a mean IC50 of 3.8 μmol/L and Hill coefficient of 0.35. Vmax was shifted from (4.0± 2.0) mV to (3.8 ± 1.6) mV. The V1/2 of inactivation curve was shifted from (- 32 ± 5) mV to (- 35 ± 4) mV and the slope factor was 31 ± 5 and 35 ± 6 before and after administration respectively. Conclusion Propofol produces significant inhibition of calcium currents in the central neurons which may partly explain the action of propofol on central nervous system.

Objective To evaluate the significance and role of ultrasound parameters in the female stress urinary incontinence (SUI). Methods The changes of the distance of bladder neck mobility,posterior urethra-vesical angle,urethral angle and residual urine volume before and after the operation of transobturator tension-free vaginal tape surgery (TVT-O) for 46 cases of female SUI (experimental group ) by ultrasound were studied,and compared with 43 normal women (control group). Results The experimental group before the operation had the greater distance of bladder neck mobility and posterior urethra-vesical angle [(14.46 ± 1.28) mm, (124.87 ±2.95)°] than the control group [(7.47 ±0.55) mm, (107.83 ±3.24)°] (P ＜ 0.01 ), but the urethral angle [( 23.61 ± 2.28 )°] was smaller than the control group [(36.24 ±2.23 )°] (P ＜ 0.01 ). There was no significant difference in the distance of bladder neck mobility, posterior urethra-vesical angle, urethral angle and residual urine volume between the experimental group after the operation and the control group (P ＞ 0.05 ). The experimental group before the operation had the greater distance of bladder neck mobility and posterior urethra-vesieal angle than those after the operation [(7.84±0.76) mm, (108.74±3.63)°] (P ＜0.01), the urethral angle was smaller than that after the operation [(34.39 ± 3.46)°] (P ＜ 0.01 ), but the residual urine volume had no significant difference (P ＞ 0.05 ).Conclusions After the operation, there are some changes in the distance of bladder neck mobility, posterior urethra-vesical angle and urethral angle for the patients with female SUI,and these parameters can be restored the normal state,and have the relatively stable effect. The residual urine volume dosen't increase after the operation. Ultrasound has the advantage in objectively assessing the severity and recovery status after the operation exclusion of mental and psychological factors.

Objective To observe the relationship between Toll-like receptor 4 (TLR4) and the sensitivity of PANC1 cells to gemcitabine (GEM),and to analyze the potential mechanism.Methods PANC1 cells were divided into GEM group,lipopolysaccharide (LPS) + GEM group and TLR4-siRNA + GEM group.GEM group was treated by GEM alone.LPS + GEM group was pretreated with 1 mg/L LPS for 4 h and then treated by GEM.TLR4-siRNA + GEM group was transfected with 100 pmol/mL TLR4-siRNA for 4 h and then treated by GEM.The untreated cells were used as the control group.MTT method was used to detect the cell proliferation.Morphological changes and apoptosis rate of the cells were examined by Hoechst33258 staining and flow cytometry,respectively.The protein expression of TLR4,phosphorylated AKT (p-AKT) and activated Caspase-3 were detected by Western blot.Results The median inhibition concentration (ICs0) of GEM in the GEM group,LPS + GEM group and TLR4-siRNA + GEM group was (8.9 ± 0.32),(14.21 ±0.95),(3.96 ± 0.27) mg/L,respectively.The IC50 in LPS + GEM group was significantly higher than that in GEM group (P ＜ 0.01),and the IC50 of GEM in TLR4-siRNA + GEM group was significantly lower than that in GEM group (P ＜0.01).Compared with that in GEM group,the cells with typical apoptotic morphological changes were decreased in LPS + GEM group,which was increased in TLR4-siRNA + GEM group.The apoptotic rate in control group,GEM group,LPS + GEM group,TLR4-siRNA + GEM group was (2.1 ± 0.3) ％,(15.1 ± 2.3) ％,(9.8 ± 1.5) ％,(22.9 ± 3.1) ％,respectively.Compared with that in GEM group,the cells apoptotic rate was significantly reduced in LPS + GEM group (P ＜0.01),which was significantly increased in TLR4-siRNA + GEM group (P ＜0.01).TLR4 protein level in the 4 groups was 0.83 ±0.08,0.81 ±0.07,0.85 ±0.07 and 0.16 ±0.03;p-AKT protein level 0.61 ±0.05,0.36 ±0.03,0.73 ± 0.07 and 0.21 ± 0.02;activated Caspase-3 protein level was 0.66 ± 0.05,0.73 ± 0.07,0.45 ± 0.04 and 0.91 ± 0.07,respectively.The expression of TLR4 and p-AKT in TLR4-siRNA + GEM group was significantly lower than that in GEM group (P ＜0.01),while the expression of activated Caspase-3 protein was increased significantly (P ＜ 0.05).Compared with the GEM group,the expression of p-AKT protein in LPS + GEM group was significantly increased (P＜0.01),and the expression of activated Caspase-3 protein was significantly decreased (P＜0.01).Conclusions TLR4 can inhibit the sensitivity of pancreatic cancer PNAC1 cells to GEM,and the mechanism is related to the activation of PI3K/AKT pathway and downregulation of activated Caspase-3.

Objective To observe the hypoglycemic mechanism of total saponins of Momordica charantia (MC) in type 2 diabetes mellitus (DM) rats. Methods Among the selected 60 male Specific Pathogen Free (SPF)rats, 8 were random- ly chosen as control group, while others were fed with high fat and high glucose diet following streptozotocin injection from caudal vein 8 weeks after to construct type 2 DM models. After the DM models were successfully built, rats were then ran- domized into five groups: DM control group (n=8), the metformin group (n=8) and three groups of total saponins of MC with different dosage (n=8 in each group). The total saponins of MC groups include DM rats administrated with total saponins of MC 100, 200, 400 mg/(kg·d) for 8 weeks, the metformin group include DM rats administrated with metformin 50 mg/(kg·d) for 8 weeks. At the end of the experiment, the fasting blood glucose and insulin were examed. At the same time, a part of pan- creas islet, liver and skeletal muscle were preserved. The pancreas islet structure, the hepatic glycogen and Glucose trans- porter 4 (GLUT4) expression were observed by electron microscope, glycogen dyeing and immunoblot respectively. Results After 8 weeks treatment, compared to type 2 DM control group, fasting blood glucose and insulin values in MC groups were reduced more obviously. However, skeletal muscle GLUT4 expression level, insulin granules and hepatic glycogen increased obviously in MC groups. Conclusion Total saponins of MC has hypoglycemic effect. It’s mechanisms maybe include pro- moting the hepatic glycogen synthesis, inhibiting the hepatic glycogen decomposition and promoting insulin sensitivity by in- creasing peripheral tissue’s GLUT4 expression.

Objective To investigate the therapeutic effect of autologous stem cell transplantation research for patients with cerebral hemorrhage sequelae under the stereotactic.Methods One hundred patients with cerebral hemorrhage from Jan.2011 to Sep.2013 in our hospital were selected and randomly divided into the experimental group (n =50) and the control group (n =50).The patients of experimental group were given autologous stem cell transplantation under the stereotactic in 6 months after cerebral hemorrhage,while the patients in control group were just given traditional treatment.At 6,7 and 12 months after cerebral hemorrhage,rate with neural function defect scale and functional independence measure(FIM) scores of the two groups were compared.Results FIM scores in the experimental group was 102.08 ± 8.28,significant higher than that in control group(95.28±8.75,P＜0.05).Functional independence measure scores in the experimental group at 7 months after cerebral hemorrhage was 13.12±4.00,significant lower than that in control group(20.40±4.33,P ＜0.05).While,there was no statistical difference at 6 months and 12 months after cerebral hemorrhage between the two groups(P＞0.05).Conclusion The therapeutic of autologous stem cell transplantation on patients with cerebral hemorrhage sequelae under the stereotactic is benefit at short term,but the long term therapy effective still needs further study.