Objective To investigate the aberrant methylation and expression of growth arrest and DNA-damage-in-ducible 45 gamma (GADD45G) gene in gastric cardia adenocarcinoma (GCA). Methods Bisulfite conversion-methylation specific polymerase chain reaction method (BS-MSP) and immunohistochemistry method were used respectively to detect the methylation status and protein expression of GADD45G in 138 GCA tumor tissues and corresponding normal tissues. Re-sults The methylation status of GADD45G distal promoter (region 1) was not detected in GCA tumor tissues and corre-sponding normal tissues. For GADD45G region 2 and region 3, the BS-MSP results of region 3 were identical to that of re-gion 2. The methylation frequency of proximal promoter and exon 1 in GADD45G island 2 (region 2 and region 3) in GCA tu-mor tissues (49.3%, 68/138) was significantly increased compared to that in corresponding normal tissues (0, P＜0.01). The methylation status of this two sites in tumor tissues was associated with TNM stage of tumors (P＜0.05). The protein expres-sion of GADD45G in tumor tissues was significantly decreased than that in corresponding normal tissues (P ＜ 0.05),and threre was a significant negative correlation with methylation status of GADD45G proximal promoter and exon 1 (rs=-0.398). Conclusion The decreased expression of GADD45G by hypermethylation of proximal promoter exon 1 of the gene may play an important role in gastric cardia adenocarcinoma.

Objective To study the role of src-suppressed C kinase substrate (SSeCKS) in the secretion of tumor necrosis factor (TNF-α) in rat pulmonary micro-vascular endothelial cells (PMVEC) induced by lipopolysaccharide (LPS).Methods Wistar rat PMVEC cultured in vitro were randomly (random number) divided into several groups (n =4) as per exposure to given dosage of LPS for different lengths of time and to different dosages of LPS for given length of time.After PMVEC exposed to 10 mg/L LPS for 1 hour (h),3 h,6 h,12 h and 24 h or 0.1 mg/L,1 mg/L and 10 mg/L LPS for 24 h,the levels of TNF-αin the supernatant of culture medium were examined by the method of enzyme linked immunosorbent assay (ELISA).Another PMVEC was pre-treated by protein kinase C (PKC) inhibitor bis-indolylmaleimide (BIM) for 0.5 h or had the transfection of SSeCKS-specific small interfering RNA (siRNA) for 48 h before 10 mg/L LPS challenge for 24 h,and subsequently the supernatant was also examined by ELISA.One-way analysis of variance (ANOVA) was employed for statistical analysis by SPSS version 10.0 to compare values among all groups.A significant difference was presumed as a probability value ＜ 0.05.Results After PMVEC incubated with 0.1 mg/L,1 mg/L and 10 mg/L LPS for 24 hours,the levels of TNF-αsecreted were (253.70 ± 23.55),(327.88 ± 37.25),(403.20 ± 36.22),respectively,which were higher than that in un-stimulated PMVEC (82.28 ± 22.56,all P =0.000).After 10 mg/L LPS challenge for one hour,the level of TNF-αin the supernatant of PMVEC raised substantially (170.11 ±49.22),peaked at the time of 6 h (404.82 ± 13.78),then persisted at a higher level until 24 h (395.67 ± 36.23) than that in un-stimulated PMVEC (84.60 ± 23.61,P =0.001,0.000,0.000,respectively).After PMVEC pre-incubated with BIM,the level of LPS-induced TNF-αdecreased obviously (200.44 ± 27.39 vs.402.28 ± 31.07,P =0.000).Compared with LPS challenged PMVEC (407.28 ± 32.64),depletion of endogenous SSeCKS in PMVEC after inhibited by SSeCKS-siRNA significantly attenuated increase in the level of LPS-induced TNF-α (195.20 ± 13.28,P =0.000).Conclusions Down-activation of SSeCKS and PKC can inhibit the secretion of TNF-αin PMVEC induced by LPS,relieving the inflammatory response of PMVEC.

Objective To investigate that p53 expression in fibroblast-like synoviocytes (FLS) and its effects on CD4+ T lymphocytes from active rheumatoid arthritis (RA) patients. Methods Human FLS was transfected with p53siRNA and cocultured with CD4+ T lymphocytes from active RA patients. The expression of osteoprotegerin and IL-6 secretion was detected in the transfected FLS. In addition, the expressions of IFN-γ, IL-17, IL-4 and CD25 as well as mRNA levels of IFN-γ RORγt, IL-17 and Foxp3 in cocuhured CD4+ T lymphocytes were also measured. Results IL-6 secretion decreased in p53-inhibited FLS while osteopro-tegerin expression was not altered, p53-inhibited FLS could significantly increase IL-17 and IFN-γ expres-sions. It also upregulated Foxp3 expression though had no effect on CD4+CD25high T lymphocytes. Conclusion p53 expression in FLS regulates Th1 and Th17 cells of RA patients, and therefore participate in the pathogenesis of RA.

Objective To observe the clinical effect of Sangpa Zhike decoction in the treatment of acute exacerbations of chronic bronchitis. Methods 180 patients with acute exacerbation of chronic bronchitis admitted to Autonomous Prefecture Hospital of traditional Chinese medicine(TCM)in Changji,Xinjiang,from August 2012 to August 2013 were enrolled. They were divided into Sangpa Zhike decoction treatment group and western medicine treatment control group by random number table,90 cases in each group. General treatments were given to the two groups. In the Sangpa Zhike decoction treatment group,additionally only oral Sangpa Zhike decoction was given(composition:Morus alba 10 g,loquat leaf 10 g,Houttuynia 10 g,honey aster 15 g,honey coltsfoot flower 6 g,Zhejiang Fritillaria 12 g,perilla 10 g,white mustard seed 6 g,Platycodon grandiflorum 10 g,Rhizoma Cynanchi Stauntonii 12 g,tangerine peel 6 g,Stemona 6 g). The ingredients were mixed in water and boiled to form a decoction,one dose daily,divided into two parts to be taken twice a day. In the western medicine treatment group,the infection was controlled by western medicine and the drugs to eliminate cough and phlegm,etc were used. The therapeutic course in both groups was 7 days. The therapeutic effect for treatment of TCM syndromes and the cough quantized integration score in the two groups were observed after treatment. Results The cure rate of Sangpa Zhike decoction treatment group was significantly higher than that of western medicine treatment control group〔70.00%(63/90)vs. 33.33%(30/90),P<0.01〕. Before treatment,the cough symptom quantization integral scores in the two groups had no statistically significance, but after treatment,the scores in the two groups were significantly lower than those before treatment. And the score in Sangpa Zhike decoction treatment group was decreased more significantly (1.66±1.12 vs. 4.36±2.32, P<0.01). Conclusion Sangpa Zhike Decoction has obvious curative effect in the treatment of acute attack of chronic bronchitis.

Objective To investigate the clinical characteristics,minimal invasive operation technique and perioperative management of lateral ventrieular tumors.Methods The clinical characteristics,image diagnosis,surgical approaches and postoperative management and surgical outcomes of 65 consecutive cases of lateral ventricular tumors were retrospectively analyzed.Results In our series,total resection was achieved in 54 cases,and subtotal resection was achieved in 11 cases.Lateral ventricular tumors were mostly found in male and in the left side.Headache caused by increased intracranial pressure was the most common clinical symptom.Ependymocytoma and astrocytic glioma are the most common pathologic diagnosis.Postoperative complications included fever (26 cases),hydrocephalus (9 cases),intraventricular hematoma (7 cases) of which 2 cases were evacuated by craniotomy,epilepsy (7 cases),wound infection (3 cases).Postoperative death was happened in 3 cases.Two of them died of respiratory failure due to postoperative epilepsy.Conclusion Early discovery of lateral ventricle tumors,meticulous operation,subtle micromanipulation and right postoperative treatment are the criticality to improve the rate of total resection of lateral ventricle tumors through microsurgical treatment and reduce postoperative complications and mortality.

Objective To study the conditions and methods of hydrodynamics-based transgene into rat regenerating liver in vivo. Methods The solution with concentration 30mg/L gene-containing plasmid was injected into rat tail veins at a speed of 2ml/s, then partial hepatectomy (PH) was performed at different times before/after injection, finally the rat (g) and regenerating liver (g) were weighed, and the liver coefficient (Lc) was calculated. Out of 15 groups which are Lc±Lc*0%, *5%, *10%, *15%, *20%, *25%, *30%, *35%, the most suitable group was chosen as correction coefficient to calculate the most appropriate volume of plasmid solution which was injected into the regenerating liver at different recovery times, and at the same time, right lobe of liver was gathered to make frozen section, then observe and quantify the positive green fluorescent protein (GFP) rate at 488 nm excitation wavelength. Results Injection of either physiological saline or empty plasmid has no significant difference compared with control (only PH performance). The appropriate time of hydrodynamics-based transgene is more than 12 hours before PH or anytime after PH. The solution volume of hydrodynamics-based transgene into liver regenerating rat after PH is rat weight (g) ×9%×1/3×corresponding correction coefficient (Trc). Both vector and target gene have effect on the time and abundance of gene expression. Conclusion Hydrodynamics-based transgene can effectively be applied to gene transfection in rat regenerating liver.

Objective To investigate the inhibitory effect of 5-fluorouracil (5-FU) on hepatocellular carcinoma (HCC) cell growth and to elucidate its potential molecular mechanism.Methods Real-time PCR analysis was conducted to determine the expression of miR-373 in HCC tissue specimens and HCC cell lines.The expression of miR-373 was also evaluated in HepG2 cells after 5-FU treatment.Western blot analysis was performed to detect the protein levels of PPP6C,a verified target of miR-373,with transfection of miR-373 mimics or 5-FU treatment.A rescue assay was conducted to investigate the cell growth in HepG2 cells by using CCK-8.Results miR-373 expression was up-regulated in both HCC tissues and cell lines.miR-373 expression depicted about 2.94-fold augment in HepG2 cells as compared to normal liver cells control (P ＜0.01).5-FU treatment led to a significant decrease of miR-373 levels (approximately 50％,P ＜0.01,48 h) and resulted in a marked increase of PPP6C protein (approximately 2.1-fold,48 h) in HepG2 cells.The overexpression of miR-373 could prevent the impact of 5-FU treatment on cell growth in HepG2 cells and CCK-8 assay showed that HepG2 cell growth was rescued approximately 81％ and 84％ at 24 h (P ＜ 0.05) and 48 h (P ＜ 0.01),respectively.Conclusion 5-FU can repress endogenous miR-373 level,which activates the expression of downstream targeted gene PPP6C,thereby exerting an inhibitory effect on HepG2 cells.

BACKGROUND: Both hydroxyapatite (HA) and large diameter TiO2 nanotubes have excellent biocompatibility, but bone-forming ability of nano-HA (nHA) deposited large diameter TiO2 nanotubes is rarely reported.OBJECTIVE: To evaluate the bone-forming ability of nHA/large-diameter TiO2 nanotube composite coating.METHODS: Large-diameter TiO2 nanotubes were prepared by anodic oxidation method, and then nHA was electrochemically deposited on the surface of TiO2 nanotubes. Preosteoblasts MC3T3-E1 were co-cultured with the nHA/large diameter TiO2 nanotube composite, pure titanium and TiO2 nanotube coatings, respectively. At 0.5, 1, 2 hours after culture, the initial cell adhesion was observed. At 1, 3, 5 day after culture, cell proliferation was assessed. At 2 days after culture, cell morphology was observed. At 3 and 7 days after osteogenic induction, intracellular alkaline phosphatase activity was detected. At 14 days after osteogenic induction, mineralization of extracellular matrix was detected.RESULTS AND CONCLUSION: (1) After 2 hours of culture, the number of adherent cells on the composite coating was significantly lower than that on the TiO2 nanotube coating (P < 0.05), but slightly higher than that on the pure titanium coating with no statistical difference. (2) After 1, 3, 5 days of culture, the cell proliferation on the composite coating was significantly lower than that on the TiO2 nanotube coating (P < 0.05), but slightly higher than that on the pure titanium with no statistical difference. (3) The cells on the pure titanium showed a spindle-shape, while those on the TiO2 nanotube coating processed filopodia. The cells on the composite coating showed polygonal shape with a larger number of filopodia. (4) The intracellular alkaline phosphatase activity of the composite coating group was significantly higher than that of the pure titanium group and TiO2 nanotube group. The trend of mineralization of extracellular matrix was ranked from high to low: the composite coating group > TiO2 nanotube group > pure titanium group. To conclude, the nHA/large diameter TiO2 nanotube composite coating not only has good biocompatibility, but also has the ideal ability to promote bone formation.

Objective To investigate the role of heparanase (HPSE) in regulating the liver regeneration after major hepatectomy in rats. Methods The HPSE small interfering RNA (siRNA) was designed and synthesized, and was transfected into the rats after 70% hepatectomy by in wwo-jetPE--Gal vector. Then immunohistochemistry was performed to determine the expression of hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF). Western blotting analysis was performed to detect the expression of matrix metalloprotein (MMP)-2 and MMP-9 protein expression. Immunostaining for VIII factor was done to observe the microvessels, and the microvessel density (MVD) was calculated. Results Compared with control group, the expressions of HGF, VEGF, MMP-2 and MMP-9 were significantly inhibited in HPSE siRNA group, and theMVD was significantly decreased (P<0. 01). Conclusion HPSE may regulate liver regeneration via promoting HGF release in ECM, inducing hepatocyte proliferation, increasing VEGF expression to accelerate angiogenesis and upregulating the expression of MMP-2 and MMP-9.