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.

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 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 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 evaluate the effect of silver needle injection therapy on rat with sports muscle injury. Methods Twenty-one healthy male Wistar rats were randomly divided into the injury group (n = 3),the silver needle group (n=12) and the control group (n=3). The expressions of bFGF and GDNF in gastrocnemius muscle tendon junction were detected on 7 d ,14 d and 28 d post-injury. Results No significant difference in the appearance of the injured tissue was found in both two groups on 7 d post-injury. The appearance of the injured tissue was better in the silver needle group than that in the control group on 14 d and 28 d post-injury. The tissue was almost normal in the therapy group on 28 d post-injury; The expression of bFGF in the therapy group was higher than that in the injury control group on 7 d and 14 d post-injury (P < 0.01). The expression of bFGF markedly decreased in the therapy group compared with the control group (P < 0.01) on 28 d post-injury. The expression of GDNF in the therapy group was higher than that in the injury control group on 7 d ,14 d and 28 d post-injury (P<0.01). Conclusion The silver needle injection therapy has the therapeutic effect on sports muscle injury reparation, which can increase the expression of bFGF and GDNF efficiently.

Objective To explore the change of serum thyroid hormone related indicators and the probability of occurrence of thyroid dysfunction abnormality in the patient with type 2 diabetes mellitus(T2DM).Methods 86 patients with T2DM and 61 age-matched and gender-matched individuals with healthy physical examination as controls were selected and detected serum FT3,FT4 and TSH by the electrochemiluminescence method Results The serum FT3,FT4 and TSH in the T2DM group were 5.09 pmol/L, 17.32 pmol/L and 2.81 mIU/L respectively;which in the normal control group were 4.99 pmol/L,17.24 pmol/L and 2.71 mIU/L respectively,the differences between the two groups had no statistical significance(P >0.05).Among 86 cases of T2DM,29 cases had the serum abnormal TSH with the abnormal rate of 33.7%,which in the control group was 14.8% with statistical difference between the two groups(P <0.05).Among T2DM patients,the TSH abnormal rate of in females was 42.1%,which was higher than 17.2% in males.Conclusion The serum thyroid hormone detection is necessary for the T2DM patients,especially female pa-tients,which is conducive to early screening,prevention and treatment.

Objective To investigate the dyslipidemia in patients with psoriasis,and the characteristics of the serum lipids in pa-tients with different types of psoriasis.Methods 332 psoriatic patients were devided into pustular psoriatic,psoriasis vulgaris and erythrodermic psoriasis based on clinical manifestations.Serum lipids of the three groups were analyzed including TC,TG,HDL-C, LDL-C,ApoA1,ApoB and Lp(a).Results The serum concentrations of TC,HDL-C,ApoA1 in patients with psoriasis were signifi-cantly lower than those in control group(P <0.05),while the concentration of Lp(a)was significantly higher in psoriasis patients compared with those in control group(P <0.05).The serum level of TC,HDL-C,LDL-C and ApoA1 in pustular psoriasis group were significantly lower than those in the other two groups(P <0.05).Conclusion The patients with psoriasis have dyslipidemia, and the serious disorder situation could be associated with psoriasis types.

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.