Objective To study the expression and its significance of Platelet-derived growth factor(PDGF) and Collagen type Ⅰ (Col- Ⅰ) mRNA in hepatic tissue in mice exposed to arsenic in drinking water. Methods Fifty mice were divided into control group, sodium arsenite group(iAs3+ group, 300 mg/L) and sodium arsenate group (iAs5+ group, 300mg/L) at random. The mice were sacrificed after 10 months for liver function test (ALT, AST and GLB examined by automatic biochemical analyzer) and pathologic examinations. Masson staining and immunohistochemical SABC methods were used. Through the result of Masson staining fibrosis semi-quantitative analysis of the fibrosis number was carried out in the per unit area. Using immuonhistochemical SABC methods, the expression of PDGF and Col- Ⅰ was detected. Results ①Serum ALT was statistically significant among control group [(36.67±3.51) U/L], iAs3+ group[(61.46±13.85) U/L] and iAs5+ group[(43.31±4.21) U/L, F= 6.56, P < 0.05], especially between the control and iAs3+ group (P < 0.05) ;Serum AST had a statistical significance among control group [(135.00 ± 20.42)U/L], iA3+ group [(510.86 ± 59.01)U/L] and iAs5+ group[(258.93 ± 22.40) U/L, F= 83.36, P < 0.05] ;Serum GLB had no statistical significance among control group [(20.86 ± 0.61)g/L], iAs3+ group [(26.94± 3.73)g/L] and iAs5+ group[(24.59 ± 5.27)g/L, F = 2.80, P < 0.05]. ②Masson staining revealed notable liver cell necrosis, regeneration, infiltration of inflammatory cells in portal area. Masson staining Semi-quantitative results showed significantly increasing size of fibrosis in model group. Compared with control group(0.069 ± 0.013) and iAs5+ group(0.143 ± 0.122), fiber hyperplasia of iAs3+ group(0.192 ± 0.108) had statistical significance (F = 1.91, P < 0.05). ③Immunohistochemistry results showed higher expression of PDGF in the portal area, sepetal and preisinus cells in model group. The heavy expression of Col- Ⅰ distributed in the surrounding of blood vessels and bile duet and along portal wall extensting to the liver in model group. Semi-quantitative results showed that PDGF among control group(202.79 ± 7.46), iAs3+ group(174.38 ± 7.71) and iAs5+ group(177.64 ± 7.81) had a statistical significance(F= 102.91, P < 0.05);Col-Ⅰ had a statistical significance among control group(200.11 ± 7.46), iAs3+ group (176.47±10.20) and iAs5+ group (177.378 ±7.948, F = 78.51, P < 0.05). Conclusions Exposed in oral drinking arsenic, mice can develop hepatic injury and hepatic fibrosis. The PDGF and Col-Ⅰ play significant effects on the process of hepatic fibrosis caused by oral drinking arsenic exposed mice.

Objective In order to understand the differentially expressed genes and explore the effects on mechanism of gene expression induced by arsenic trioxide. Methods The mRNA was isolated from human HepG2 cells treated with arsenic trioxide( 5?mol/L ) and DMSO, respectively, then cDNA was synthesized. After restriction enzyme Rsa Ⅰ digestion, small sizes cDNA were obtained. Then tester cDNA was subdivided into two portions and each was ligated with different cDNA adaptor. After tester cDNA was hybridized with driver cDNA twice and underwent nested polymerase chain reaction (PCR) twice, the DNA fragment was subcloned into T/A plasmid vectors to set up the subtractive cDNA library. Amplification of the library was carried out with E. coli strain JM109. The cDNA was sequenced and analyzed in GenBank with Blast search after colony PCR. Results The forward subtracted cDNA library from HepG2 cell line induced by arsenic trioxide was successfully constructed. The sequencing analysis showed that there were eight clones contained ferritin H(L) chain in the library. Conclusion Arsenic trioxide can induce the up expression of ferritin H(L) chain protein in HepG2 cells, indicated that the ferritin H(L) chain may play certain role in the mechanism of anti-arsenical cytotoxicity in liver.

Objective To screen the proteins binding with HBV X-promoter and to investigate their potential role in the regulation of replication and expression of HBV DNA. Methods By using HBV X biotinylated promoter DNA as the selective molecule, the T7 select human liver cDNA library was biopanned and positive clones were selected. After screening, positive plaques were performed to amplify for inserted DNA fragment and cloned into pGEM-Teasy vector. Thirty positive plaques were chosen for DNA sequencing. Results Five kinds of known and nine kinds of novel cDNA sequences were obtained. The 5 kinds of known ones are human serum albumin, NADH dehydrogenase subunit 4, prokininase, ubiquitin specific protease 10, and testis enhanced gene transcript. Conclusion The binding proteins of HBV Xp DNA were identified by phage display. The results suggest that this approach provides a new search tool for the study of replication and expression mechanism of HBV DNA.

Objective To clone and bioinformatically analyze the full-length sequence of F protein binding protein 1 (FBP1). Methods Yeast two-hybrid system was employed to obtain an unknown gene and named it as FBP1.The coding sequence of FBP1 was cloned using molecular biological techniques. Results The coding sequence of FBP1 was cloned successfully. Conclusion FBP1, a cellular protein binding to F protein of HCV, plays an important role in the interaction of virus protein and host cell protein.

Objective To study the feasibility and indication of twice chemonucleolysis with collagenase for the treatment of lumbar disc herniation.Methods Eighty two patients of lumber disc herniation were treated with twice collagenase chemonucleolysis.All patients were followed up for 3 to 12 months and then the clinical results were assessed according to the Macnab criteria retrospectively.Results Eighty two cases were followed up from 3 to 12 months postoperatively.Fifty one cases were excellent,13 cases good,8 as fair and 10 were poor.The rate of excellent plus good reached 78%,the effective rate was 88%.Conclusion Twice chemonucleolysis with strict indications together with prevention of allergic reaction before,during and after the operation;is safe and effective for treating lumbar disc herniation.(J Intervent Radiol,2007,16:258-259)

Objective To explore the Clinicopathological characteristics of patients with primary adenosquamous and squamous carcinoma of stomach. Methods The clinical data of 12 cases of primary squamous and adenosquamous carcinoma of the stomach were reviewed retrospectively, and the immunohistochemical staining of CK17 and CKI8 protein were performed in primary gastric adenosquamous carcinoma. Results Primary adenosquamous and squamous carcinoma of the stomach accounted for 0.28% of all the 4352 patients with gastric cancer during the same period. Of the 12 patients, 10 were adenosquamous carcinoma and other two were squamous carcinoma. There were 10 males and 2 females in this group, with their mean age being 65 years. The main clinical presentation included epigastric pain and discomfort, followed by hematemesis and melena. The definite diagnosis rate was 33% (4/12) by gastroscopy and biopsy before operation. The tumors were less than 5 cm in diameter in 3 patients, and more than 5 cm in 9 patients. The surgical procedure was radical resection in 8 patients and palliative resection in 4 patients. There were 1 case of stage Ⅰ, 5 cases of stage Ⅲ, 6 cases of stage Ⅳ. 10 patients died of tumor recurrence and metastasis within 2 years after operation, one died of other unrelated disease, and one was alive for more than 5 months. The component of both adenosquamous and squamous carcinoma were more than 30% in 4 patients with adenosquamous carcinoma who underwent palliative resection and died within 6 months after operation. Conclusions Primary adenosquamous and squamous carcinoma of the stomach were rare, and had specific clinicopathological characteristics. Having both biological behaviours of adenocarcinoma and squamous carcinoma may lead to poor prognosis in adenosquamous carcinoma of stomach.

Objective To study the low dose hyper-radiosensitivity in human lung cancer cell line A549,and its possible mechanisms.Methods Exponentially growing A549 cells were irradiated with 60Co γ-rays at doses of 0-2 Gy.Together with flow cytometry for precise cell sorting,cell survival fraction was measured by mean of conventional colony-formation assay.ATM1981 Ser-P protein expression was examined by Western blot.Apoptosis was identified by Hoechst 33258 fluorescent staining,and Annexin V-FITC and propidium iodide staining flow cytometry.Cell cycle distribution was observed by flow cytometry.Results There was an excessive cell killing per unit dose when the doses were below about 0.3 Gy,and the cells exhibited more resistant response at the doses between 0.3 and 0.5 Gy,the cell survival fraction was decreased as the doses over 0.5 Gy.The expression of ATM1981Ser-P protein was first observed at 0.2 Gy,followed by an increase over 0.2 Gy,and reached the peak at 0.5 Gy(compared with 0.2 Gy group,t=7.96,P＜0.05),with no further increase as the doses at 1.0 and 2.0 Gy(t=0.69,0.55,P＞0.05).24 hours after irradiation,part cells presented the characteristic morpholos4cal change of apoptosis,and the apoptosis curve was coincident with the dose-survival curve.Compared with the control group,the cell cycle had no change post-irradiation to 0.1 and 0.2 Gy.G2/M phase arrest was manifested at 6 and 12 hours post-irradiation to 0.3,0.4 and 0.5 Gy(t=2.87,2.88,4.92 and 3.70,3.12,8.11,P＜0.05),and the ratio of G2/M phase was decreased at 24 hours post-irradiation(t=3.87,4.77,3.01,P＜0.05).Conclusions A549 cells displays the phenomenon of hyper-radiosensitivity(HRS)/induced radioresistance(IRR).The model of cell death induced by low dose irradiation is mainly apoptosis.The activity of ATM and cell cycle change might play an important role in HRS/IRR.

Objective To explore the effectiveness of sandplay therapy combined with family counseling in children with school phobia and its influence of child′ self-consciousness. Methods Integrative sandplay therary with family consulting were used to treat 28 patients with school phobia regularly for 2 months. Sandplay and family consulting therapy were given once a week for 45 minutes . Clinical outcomes were assessed using CGI-GI and Piers-Harris children′s self-consciousness scale before and after treatment as well as 3 months posttreatment. Results Overall response rate was 85%. In addition, the physical appearance and characteristic factor before and after treatment were no significant difference (P>0.05). The rest of the various factors and total score compared with pre-treatment significantly improved (P<0.05). After treatment for 3 months, every factor in self-consciousness of children and total score were no significant difference (P>0.05). Conclusion Integrative sandplay therapy with family counseling has better and long-lasting treatment effect to self-consciousness of children with school refusal.

Aim To investigate the effects of silencing MALAT1 gene on cell proliferation inhibition and apop-tosis induced by Melittin in human hepatocellular car-cinoma HepG2 cells. Methods The inhibitory rate of cell proliferation treated with Melittin in HepG2 cells was examined by MTT assay. Apoptotic rate was detec-ted by flow cytometry. The MALAT1 expression level in HepG2 cells was measured by qPCR. Specific siR-NAs were utilized to silence MALAT1 expression. The rates of cell proliferation inhibition and apoptosis in HepG2 cells treated with siRNA and Melittin were compared with those of Melittin alone. Results Melit-tin significantly suppressed the growth of HepG2 and induced cell apoptosis in a dose-dependent manner. Compared with normal liver cell lines, MALAT1 was highly expressed in HepG2 cells ( P<0. 05 ) . The ex-pression of MALAT1 in HepG2 cells was inhibited by Melittin, and the inhibitory rate increased with the in-crease of concentration. The rates of cell proliferation inhibition and apoptosis in HepG2 cells treated with siRNA and Melittin were significantly higher than those treated merely with Melittin. Conclusion Melittin can reduce the expression of MALAT1 and silencing MALAT1 can effectively promote proliferation inhibi-tion and apoptosis in HepG2 cells induced by Melittin.