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 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 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 study the effects of total flavonoids of Astragalus on apoptosis of bovine retinal capillary pericytes(BRPs) under high glucose.Methods: The third generation of nearly symphysic bovine retinal vessel pericytes cultivated in vitro were divided into normal control group,high glucose group,and Astragalus total flavonoids groups(0.25,0.5,1.0,2.0 mg/ml) at random.After being incubated for 6 days,apoptosis of BRPs were detected by TUNEL method.TBA method was used to detect the contents of MDA.Xanthine oxidase method was used to detect the SOD activities.Results: Compared with high glucose group,the apoptosis,MDA contents,SOD activity,MDA content/SOD activity of BRPs reduced markedly in total flavonoids of Astragalus groups(0.5,1.0,2.0mg/ml)(P

70% for 9 patients;the positive rate was between 30-70% for 4 patients;the positive rate 0.05). Conclusion Tamm-Horsfall detection of RBC in urine could be used as one of the Identification indicators for DN hematuria, and it can provide a basis for diagnosis and treatment of DN.

Objective To study MR imaging manifestations of the mesorectum and mesorectal fascia. Methods 100 cases were divided into five groups according to the age. All MR images were retrospectively observed and analyzed by two radiologists.The observing contents included: ①exhibition of the mesorectal fascia, ②the signal intensity of the mesorectum and the pelvic fat and ③the thickness ratio of the mesorectum and the subcutaneous fat in different ages. All data were dealed with SPSS 10.0 software. Results The showing rate of the anterior, posterior, left and right mesorectal fascia respectively were 77, 100, 91 and 93 by MR imaging. The signal intensity of the mesorectum(550.8843) was significantly higher than that(469.8693) of pelvic fat in all five groups(P﹤0.05) and the signal difference between the mesorectum and the pelvic fat could be detected by naked eye. The thickness of the mesorectum and the subcutaneous fat had no association with ages.Conclusion MRI can display the mesorectum and mesorectal fascia clearly. The signal intensity of the mesorectum is significantly higher than that of pelvic fat in five groups. The thickness of the mesorectum and the subcutaneous fat have no association with ages.

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.

The primary function of optical in vivo imaging technique is tracing and detecting the action and expression of maker cells,maker microorganisms and maker molecules.This technique makes possible the noinvasive study of biological events continuously with high sensibility,simplicity and high speed.It has been applied in numerous research fields.This paper presents a review of principle and application of this technique.