Objective To investigate the effects and possible mechanism of action of inhibiting hepatitis B virus X protein (HBx) expression on liver cancer metastasis. Methods The suppression of HBx expression in MHCC97H cells was performed by siRNA interference technique, and the effects of HBx suppression on the metastasis of MHCC97H cells were detected by Matrigel invasion assays and in a lung-metastasis mouse model. The expression levels of related epithelial-mesenchymal transition (EMT) and apoptosis proteins were examined by Western blotting. Results Introduction of HBx-siRNA into MHCC97H cells inhibited the expression of HBx and the ability to metastasize,downregulated the expression of Twist and N-cadherin, and upregulated E-cadherin expression. These changes resulted in inhibiting EMT of MHCC97H cells. Meanwhile, apoptosis involved in the Twist-P53 pathway was also found. Conclusions Inhibiting expression of HBx can decrease the metastatic a-bility of MHCC97H cells by changing EMT and inducing apoptosis.

Objective To apply the hydrocolloid dressings and hydrocolloid dressings combined GreenCream Dressing for central venous catheterization fixing, and to explore the effect of hydrocolloid dressings combined GreenCream Dressing in the prevention of venous catheter bacterial colonization and bacterial infection. Methods 470 patients who underwent the Inferior vena cava catheter were divided into 230 patients in the control group and 240 patients in the experimental group. The control group was fixed with hydrocolloid dressings after central venous catheter, and the experimental group was fixed with hydrocolloid dressings combined GreenCream Dressing after central vein catheter. The measurements included catheter bacterial colonization, catheter-related infections (CRIs) and catheter related blood stream infections (CR-BSIs), pathogenic bacteria colonization of the skin. At the same time, the skin safety was also confirmed. Results In the control group, 230 cases were retained for 1 419 catheter-days, and 240 cases in the experimental group were retained for 1 675 catheter-days. Compared with hydrocolloid dressings, hydrocolloid dressing combined GreenCream Dressing could reduce the incidence of CRIs from 1.8‰(3/1 675) to 0.7‰(1/1 675), and CR-BSIs from 2.4‰(4/1 675) to 0.7‰(1/1 675) respectively, with the statistically significant (χ2=6.39, 95%CI 1.30-31.41, andχ2=6.21, 95%CI 1.56-40.82;P<0.05). The results of bacterial colonization, CRIs and CR-BSIs showed that the most common bacteria were Staphylococcus and fungi. At the same time, compared with the hydrocolloid dressing, hydrocolloid dressing combined GreenCream dressing could reduce the incidence of skin pathogenic bacteria colonization, from 41.74%(96/230) to 28.33%(68/230),with the statistically significant (χ2=9.29,P=0.00);There was no difference between the two groups in the field of the incidence of abnormal skin manifestation (χ2=1.23, P=0.30), showing a good safety. Conclusions Hydrocolloid dressing combined GreenCream Dressing would be more effective to prevent bacterial colonization and bacterial infection of central venous catheter in department of neurosurgery.

Objective To investigate the effect of hydrogen peroxide (H2O2) pretreatment on free mitochondrial cyto-chrome c (Cyt-c) release in mitochondrial levels, and reveal the mechanism of the ischemic preconditioning (IPC) on the ischemia reperfusion (IR) injury thereof. Methods The rat liver mitochondria was isolated and made free mitochondria. Free mitochondria were divided into 5 groups:control group (C) and different concentrations of Ca2+groups (12.5, 25, 50 and 100 μmol/L). The levels of Cyt-c and second mitochondria-defived activator of caspase (Smac) were detected after 10 min stimulation of free mitochondria. The free mitochondrial IPC reperfusion model was made and divided into seven groups:C group, IR group and different concentrations of H2O2 groups (2 μL H2O2 in 200 μL system respectively, final concentration of 1, 2, 5, 20 and 100 μmol/L respectively). 100 μmol/L Ca2+was used again on the simulation of IR group. The level of Cyt-c release was detected. The changes in the activity of cardiolipin were detected in IR group and H2O2 (1 and 2 μmol/L of final concentration) groups.Results Compared with C group, there were significantly higher levels of Cyt-c and Smac emission in 25, 50, and 100 μmol/L Ca2+groups (P<0.05). Compared with IR group, there was significantly decreased level of Cyt-c emission in H2O2 (1 and 2 μmol/L) groups (P < 0.05). The activity of cardiolipin was changed when reducing release of Cyt-c. Conclusion Cyt-c was bonded with cardiolipin more closely when the low concentration of H2O2 pretreatment in mitochondria. There was a lower level of Cyt-c emission in mitochondria after stimulation with high concentration of Ca 2+(100 μmol/L Ca2+). The blocking apoptotic pathway plays a key fact in the effect of IPC on IR injury.

Objective To investigate whether human breast milk may bind to hepatitis B surface antigen (HBsAg) and its characteristics.Methods Breast milk samples from five women with negative HBsAg and hepatitis B surface antibody (anti-HBs) at one to two months post delivery were fractioned into cream and skimmed milk by centrifugation.The human breast milk and each fraction as well as cow and goat milk samples,served as controls,were separately incubated with highly purified yeast recombinant HBsAg,followed by determination of their binding capability to HBsAg by enzyme linked immunosorbent assay (ELISA) and the inhibition rate for binding of HBsAg to anti-HBs by quantitative chemiluminescence microparticle immunoassay.After boiled for 1 min or pasteurized in 65 ℃ for 30 min,the thermal stability of the active components of milk was detected.One-way ANOVA and SNK tests were performed for statistical analysis.Results The operative concentration of HBsAg was 0.1 μg/ml.Breast milk from all five women showed significantly better binding capability to HBsAg than cow or goat milk (1.306±0.300 vs 2.157±0.150 and 2.232±0.093,F=34.303,P＜0.01).The quantitative experiments showed that the inhibition rate of human breast milk was higher than that of the control group [(74.26± 17.26)％ vs (0.00±5.50)％,F=57.806,P＜0.01].The binding ability to HBsAg of skimmed milk was comparable with that of whole milk,indicating milk protein(s) played critical roles in binding to HBsAg (0.877 ± 0.486 vs 0.513 ± 0.069 and 0.376 ± 0.146,F=44.475,P＜0.01).After boiled for 1 min or Pasteurization,the binding ability to HBsAg of whole breast milk remained,but that of skimmed milk went down (F=16.598,P＜0.01).Both whole breast milk and skimmed milk could inhibit the binding of HBsAg to anti-HBs (F=278.341 and 269.408,both P＜0.01).Conclusions The inhibition of binding to HBsAg by human breast milk indicates that human milk may interact with HBsAg.The active components mainly exist in milk proteins and are thermal stable.

Objective:To investigate the effect of DNA vaccine expressed Helicobacter pylori(Hp) oipA on protecting against Hp infection.Methods:The ORFs of Hp oipA had been inserted into the eukaryotic expressing vector pVAX1 and SGC-7901 cells had been transfected with recombinant plasmid pVAX1-oipA. The expression of oipA had been detected by RT-PCR and ELISA. After extracted and purified, pVAX1-oipA had been injected into BALB/c mice through muscles of right leg one time each week for three weeks(100 ?g each mouse). pVAX1 blank and normal saline had been used as the control groups. Titer of antibodies had been detected by ELISA two months after the last immunization. Based on the confirmation of immunological response in the pVAX1 groups, mice had been given orogastric challenged with live Hp Sydney strain three times(0.5?108/0.5 ml each mouse). Four weeks after challenge, mice had been sacrificed. Histological change and the colonization of Hp in the gastric mucosa had been detected by urease test, the culture of Hp, and electronic microscopy.Results:SGC-7901 cells transfected with pVAX1-oipA had expressed corresponding production at the level of transcription and translation. Immunized mice had been induced anti-oipA antibodies. After challenged with bacterium, as contrast to immunized mice groups injected with pVAX1-oipA, pVAX1 blank, normal saline, the positive rate of urease test of gastric mucosa was 0(0/10), 90%(9/10), 100%(5/5)respectively,the positive rate of cultures of Hp was 20%(2/10), 90%(9/10), 100%(5/5)respectively. Histological findings: the different degree of erosion had been observed in control group, but 80%(8/10)of gastric mucosa were normal in immunized mice.Conclusion:oipA DNA could induce effective immune response in protection against Hp infection.

AIM: To investigate the influence of laminin (LN) on the expression of bcl-2 in rabbit cornea endothelial cells. METHODS: Cultured rabbit corneal endothelial cells were divided into three groups: normal control group, LN group and negative control group. Immunohistochemical technique and ELISA were used to measure the staining and A value to assess the levels of bcl-2 expression. RESULTS: Bcl-2 expression score showed that LN group had a strong positive expression. Control group only showed a weak expression. Negative control group showed a negative expression by immunohistochemical staining. The mean A value of each group were 1.21?0.18 (LN group), 1.05?0.14 (normal control group) and 0.04?0.01 (negative control group). CONCLUSION: LN promotes bcl-2 gene expression in rabbit cornea endothelial cells, and protects the cells from apoptosis.

Objective:To investigate the effect on the expression of Slug for the trasfection of miR-200c combined with the research on the ability of migration of breast cancer cell BT549.Methods:Chemically synthesized miR-200c mimic was trasfected into BT549 cells,which have high metastatic potential.The effect on the ability of migration of breast cancer cell BT549 for the transfection of miR-200c was analyzed by Transwell migration assay and Wound healing assay.The expression of Slug and E-cadherin mRNA was detected by Real-time PCR.The expression of Slug protein was detected by Western blot.Results:Transfection with miR-200c mimic significantly down-regulated the expression of Slug as compared with the control group (P<0.05).BT549 cell tranfected with miR-200c mimic had lower levels of migration capacity than cells in the control group (P<0.05).Conclusion:miR-200c inhibits Epithelial-mes-enchymal transition by suppressing Slug leading to down-regulation of migration capacity of breast cancer cell BT549.

Objective:To explore the effect of ionizing radiation on apoptosis of lung cancer H460 cells after ATRX was silenced by RNAi and its mechanism.Methods:The lentivirus expression vectors targeting ATRX were transfected into the 293T cells,and the lung cancer H460 cells were infected with lentivirus twice,and the ATRX silenced cell model was obtained after puromycin positive screening,then they were named as sh-ATRX1-H460,sh-ATRX2-H460,and sh-ATRX3-H460 cells;the sh-control-H460 cells were regarded as control cells.The cells were divided into sh-control-H460 group and sh-ATRX3-H460 group,accroding to the silencing results and were irradiated by 0,2 and 8 Gy X-rays.The expression levels of ATRX,poly(ADP-ribose) polymerase 1(PARP1),and caspase-3 proteins were measured by Western blotting method;the apoptotic rate was measured by flow cytometry and AnnexinⅤ-FITC/PI kits.Results:The lung cancer cell model of sh-ATRX3-H460 silenced by ATRX was obtained successfully.After 2 and 8 Gy X-ray irradiation,compared with before irradiation,the expression level of ATRX protein in sh-control-H460 group was increased,while there was no expression of ATRX protein in sh-control-H460 group;compared with before irradiation,the apoptotic rates of cells in two groups were increased(P<0.05 or P<0.01);the apoptotic rate in sh-ATRX3-H460 group was significantly higher than that in sh-control-H460 group after 8 Gy X-ray irradiation (P<0.01).The expressions of cleaved PARP1 in the cells in both two groups after 2 Gy and 8 Gy X-ray irradiation were increased and showed similar rule.The expression level of procaspase-3 protein in sh-control-H460 group had little change,and it was increased significantly in sh-ATRX3-H460 group after 8 Gy X-ray irradiation.Conclusion:ATRX silencing can be achived by RNAi,then the silencing could increase the apoptosis induced by irradiation and its mechanism may be related to the PARP1-caspase-3 pathway.

Objective To discuss the diagnostic value of the examination of serum NAG, AFU, PAB, LAP, ASTm, GLDH,ADA and AFP in patients suffering from liver diseases.Methods Serum of 274 hepatitis cases and 30 healthy cases are examined with auto biochemical analyzer and analyzed statistically.Results The mean values of LAP, ASTm, GLDH, ADA and AFU in acute hepatitis patients are higher than health′s significantly, AUC of AFU,LAP and ASTm are 0.842,0.816 and 0.782 separately, positive rate of AFU,LAP and ASTm are 84.2%,95% and 80% separately; The mean values of ADA、AFU and NAG in liver cirrhosis patients are higher than health′s significantly while the mean value of PAB is lower significantly, AUC of ADA is 0.689, positive rate of ADA is 89.5%; The mean values of ADA and NAG in severe hepatitis patients are higher than health′s significantly while the mean values of PAB and AFU are lower significantly, AUC of PAB and AFU all is 0.861, positive rate of PAB and AFU is 100% and 52.1%; The mean values of LAP,AFP,NAG,ADA and AFU in liver cancer patients are higher than health′s significantly while the mean value of PAB is lower significantly, AUC of LAP and AFU is 0.697 and 0.653 separately, positive rate of LAP and AFU are 74% and 79.5% separately.Conclusions AFU、LAP and ASTm are valuable markers for diagnosing of acute hepatitis, ADA is a valuable marker for diagnosing of liver cirrhosis, PAB and AFU are valuable markers for diagnosing of severe hepatitis, LAP and AFP are valuable markers for diagnosing of liver cancer.

Objective To explore the effect of electroacupuncture at Quchi (LI11), Zusanli (ST36) on differentiation of neural stem cells after cerebral ischemia-reperfusion in rats. Methods Thirty-six male Sprague-Dawley rats were randomly divided into sham group (n=12), model group (n=12) and electroacupuncture group (n=12). The latter two groups were occluded the left middle cerebral arteries for 90 min-utes and reperfused. The electroacupuncture group received electroacupuncture at Quchi and Zusanli acupoints for 21 days. They were evalu-ated with modified Neurological Severity Scores 7, 14 and 21 days after electroacupuncture. Their infarct volumes were tested with MRI T2WI 21 days after electroacupuncture, while the differentiation of neural stem cells was observed with double-immunopositive BrdU/Dcx and BrdU/NeuN. Results Compared with the model group, the neurological deficits score improved in the electroacupuncture group in all the time points (P<0.05). The infarct volumes decreased in the electroacupuncture group (P<0.05), with less number of BrdU+/Dcx+cells in subventricular zone (P<0.001) and more number of BrdU+/NeuN+ cells in peri-infarct cortex (P<0.001) 21 days after electroacupuncture. Conclusion Electroacupuncture at Quchi and Zusanli acupoints can improve neurological function and decrease the infarct volumes in rats after cerebral ischemia-reperfusion, which may be associated with promoting differentiation of neural stem cells to neurons.