Bile Duct Neoplasms ; genetics ; pathology ; Bile Ducts, Extrahepatic ; Cholangiocarcinoma ; genetics ; pathology ; Female ; GTP-Binding Protein gamma Subunits ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Objective To investigate the effects of Dishevelled (DVL) on apoptosis of diffuse large B-cell lymphoma (DLBCL) cell line OCI-Ly10, and to explore its possible mechanism. Methods Lentivirus plasmid overexpressing DVL2 was constructed, and after virus was packaged, it was transfected into OCI-Ly10 cells. Flow cytometry was used to detect the apoptosis rate of OCI-Ly10 cells with or without the stimulation by TNF-α recombinant protein. Then the gene expression of anti-apoptotic genes, GADD45β and A20, in NF-κB pathway was detected by RT-PCR. Results The virus was sucessfully transfected into OCL-Ly10 cells which overexpressed DVL2. The apoptosis rate of OCL-Ly10 cells overexpressing DVL2 without the stimulation by TNF-α was increased compared with that of the negative control group [(15.46 ±2.37) % vs. (11.72±3.53)%, P=0.03], the A20 mRNA expression level was decreased compared with that of the negative control group [(0.66 ±0.01) vs. 1, P=0.04], and the relative expression level of GADD45β mRNA was not significantly decreased compared with that of the negative control group [(0.79 ±0.15) vs. 1, P=0.642]. The apoptosis rate of DVL2 overexpression OCI-Ly10 cells stimulated by TNF-α was significantly higher than that of the negative control group treated by TNF-α [(22.78±4.56)%vs. (12.79±2.89)%, P=0.007]. The gene expression of A20 and GADD45β in DVL2 overexpression cells stimulated by TNF-α was significantly increased, however, the magnitude of increase in DVL2 overexpression cells was less than that in the negative control group treated by TNF-α [A20: (3.75 ±0.14) times vs. (6.89 ±0.10) times, P=0.008; GADD45β:(4.750±0.21) times vs. (6.14±0.08) times, P=0.03]. Conclusion DVL can promote the apoptosis of OCI-Ly10 cells, and its mechanism may be related with anti-apoptotic genes that inhibits its downstream via NF-κB pathway.

OBJECTIVETo investigate PTCH gene mutations in odontogenic keratocysts (OKC).

METHODSPCR-SSCP and DNA sequencing were used to analyze the PTCH gene mutations in 12 OKCs, including 10 sporadic and 2 nevoid basal cell carcinoma syndrome (NBCCS) associated OKC.

RESULTSFour mutations were identified in 4 cysts, among which two germline mutations were associated with NBCCS and 2 somatic mutations were in 2 unrelated sporadic cases. In addition, eight previously reported polymorphisms in the PTCH gene were also found in 10 cases.

CONCLUSIONThe present study indicated that both sporadic and NBCCS-related OKCs could carry PTCH gene mutation. Thus, mutational inactivation of PTCH gene may play a significant role in the pathogenesis of OKC.

Basal Cell Nevus Syndrome ; Female ; Humans ; Mutation ; Odontogenic Cysts ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Single-Stranded Conformational ; Receptors, Cell Surface ; Sequence Analysis, DNA

Objective To evaluate the success rate, safety and efficacy of endoscopic retrograde cholangiopancreatography (ERCP) after Billroth Ⅱ gastrectomy. Methods Data of 75 patients with biliary disease after Billroth Ⅱ gastrectomy, who underwent ERCP from January 2007 to November 2009, were retrospectively analyzed. Results In 75 patients, afferent loop intubation was achieved in 69 (92%) and selective cannulation of bile duct were successful in 68 (68/69, 98. 5%). Diagnostic procedures were carried out in 3 patients, and therapeutic in 65 others, which included EST plus stone removal and ENBD in 16, ERBD in 19, EMBE in 18 and EBD plus stone removal and ENBD in 12. Afferent loop perforation occurred in 1 patient (1.3%) and was treated surgically, and 2 acute pancreatitis (2. 6%) were treated conservatively.There was no complication of bleeding. Conclusion ERCP after Billroth Ⅱ gastrectomy is safe and efficiency for biliary disease.

Objective To investigate the effects of the folic acid on human T lymphoid leukemia cell line CEM cells. Methods 1. MTT method was used to detect the proliferation of CEM cells co- cultured with folic acid of different concentrations and time;2. E-xamine the changes of morphology by light microscopy with Giemsa stain;3. Detect the percentage of apoptosis and cell cycle distribution as well as the expression of the apoptosis protein(Bcl- 2,C- myc) by flow cytometry;4. Detect DNA fragments by Agaiose elec-trophoresis;5. Detect the influence of folic acid to the anticancer effects of methotrexatc(MTX) by MTT methods. Results 1 Folic acid could inhibit the proliferation of CEM cells, and the optimal inhibitive concentrations range from 0. 4 ? 10-4 ?g/L to 3. 0 ? 10 -4 ?g/L,the inhibition rate was about 30% - 40% ; 2. Co - cultured with folic acid at 24,48, 72 hours, examined by light microscopy with Giemsa stain, apoptosis cells were found in all study groups but the higher apoptosis rate was found co - cultured with folic acid at concentration of (0.4 - 3.0) ? 10-4?g/L;3.The highest apoptosis rate was 6. 19% found at the concentration of 3 ? 10-4 ?g/L, but the cell cycle distribution had no statistical difference with control group, the expression of apoptosis related protein Bcl - 2 and C-myc was decreased;4 DNA was extracted from CEM cells co - cultured with 0.4? 10 -4 ?g/L and 3 ? 10-4 ?g/L folic acid for 48 hours. UNA ladders were visible by agarose electrophoresis of DNA fragments; 5. Folic acid did not affect the antitumor effect of MTX at the concentration from 0 2? 10-4 ?g/L to 12.0?10-4 ?g/L. Conclusion Folic acid may suppress proliferation and induce apoptosis of CEM cell

OBJECTIVETo preliminarily investigate the influence of hypoxia on human umbilical vein endothelial cells (HUVECs), and the effect of Ginkgo biloba extract 50 (GBE50) on it.

METHODSFlow cytometry, TUNEL, RT-PCR, Western blot, etc. were applied, to study the effect of hypoxia and GBE50 on endothelial cells.

RESULTSAfter being interfered by hypoxia for 24 h, the levels of reactive oxygen species (ROS) in HUVECs and the apoptotic rate either in the early or in the late stage significantly increased, and accompanied with the increased levels of endothelin-1 mRNA (ET-1) and endothelial oxide synthase (eNOS) protein. However, when HUVECs were pretreated with GBE50 (25 [microg/ml) 4 h before hypoxia, the apoptotic rate in the early or late stage and expression of ET-1 mRNA significantly decreased (P < 0.05), and the heightened ROS level and eNOS expression partially decreased (P > 0.05).

CONCLUSIONHypoxia can induce endothelial dysfunction, which could be partially or significantly reversed by GBE50, it shows a certain protective effect on hypoxia induced endothelial dysfunction.

Blotting, Western ; Cell Hypoxia ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Endothelial Cells ; drug effects ; metabolism ; Endothelin-1 ; biosynthesis ; genetics ; Endothelium, Vascular ; drug effects ; metabolism ; physiopathology ; Free Radical Scavengers ; pharmacology ; Ginkgo biloba ; chemistry ; Humans ; Plant Leaves ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Reactive Oxygen Species ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Psychrotrophs were isolated by using four media from low-temperature sewage of sewage treatment plant in Urumqi, Xinjiang. Totally, 154 strains were obtained including 12 filamentous fungi, 46 yeasts, 6 actinomycetes and 90 bacteria. The results of tolerance tests of the isolates to salt, phenol and SDS, and enzyme producing characters of amylase, proteinase and esterase were shown. Then 60 bacterial strains were chosen for 16S rRNA gene sequencing and analysis. The blasting results showed that the strains were assigned to 13 recognized genera , and the Strain 39 exhibited 96.6% similarity to Acinetobacter lwoffii(DSM2403), indicating that it might be a novel species. These results suggested that there were a lot of psychrotrophs and rich bacterial diversity in low-temperature sewage. In addition, which maybe an important and potential library of microbial resources.

OBJECTIVETo evaluate the effect of arthroscopic debridement for knee osteoarthritis.

METHODSThe medical records of 56 patients (56 knees) who underwent arthroscopic debridement procedures for knee osteoarthritis from 2000 to 2004 were reviewed. Among the patients, 15 patients were male and 41 patients were female,ranged in age from 39 to 76 years, with an average of (55.23 +/- 10.26) years. The duration of the disease ranged from 1 to 25 months, with an average of 5.75 months. The chief symptoms were pain, swelling and dysfunction of the knees. According to Kellgren-Lawrence classification grade, 17 patients were grade I ,39 patients were grade II. Lysholm score was used to evaluate the postoperative effects, and the statistics analysis was carried out with SPSS 13.0.

RESULTSAll the patients were followed up more than 5 years. Three patients underwent total knee replacement respectively at 5.5, 7.8 and 8.3 years after the arthroscopic debridement, the average Lysholm score of other 53 patients increased from (42.40 +/- 6.78) preoperatively to (75.53 +/- 8.23) postoperatively. There were significant difference between preoperative score and postoperative score (t = -22.62, P < 0.01).

CONCLUSIONThe good midterm follow-up results of arthroscopic debridement for knee osteoarthritis are related to the appropriate indications selections, limited debridement, good patient education, systemic rehabilitation and chondroprotective agents.

Adult ; Aged ; Arthroscopy ; Debridement ; Female ; Follow-Up Studies ; Health Education ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; pathology ; surgery

BACKGROUNDTo investigate whether Borna disease virus (BDV) infection is related to the schizophrenic patients from China.

METHODSA reliable Western-blot method for detection of BDV-p24 antibody was established by adjusting the reaction conditions of BDV-p24 recombinant protein and specific antibodies. The sera of schizophrenic patients and normal controls from Heilongjiang Province were screened for specific BDV-p24 antibody by this method, and the BDV-p24 antibody positive sera were confirmed by the Western-blot method with sera-GST protein absorption.

RESULTSTen of 116 (8.6%) schizophrenic patients were found to be positive for BDV-p24 specific antibody, while no BDV-p24 specific antibody was found in sera of normal controls.

CONCLUSIONSThe results demonstrate that the Borna disease virus infection also exists in China, and the infection is possibly associated with schizophrenia in some way.

Antibodies, Viral ; blood ; Blotting, Western ; Borna disease virus ; isolation & purification ; Humans ; Schizophrenia ; virology ; Viral Proteins ; immunology

BACKGROUNDIt is important to identify the multiple sites of leptin activity in obese women with breast cancer. In this study, we examined the effect of exogenous human leptin on heat shock protein 70 (HSP70) expression in MCF-7 human breast cancer cells and in a breast carcinoma xenograft model of nude mice.

METHODSWe cultured MCF-7 human breast cancer cells and established nude mice bearing xenografts of these cells, and randomly divided them into experimental and control groups. The experimental group was treated with human leptin, while the control group was treated with the same volume of normal saline. A real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assay was developed to quantify the mRNA expression of HSP70 in the MCF-7 human breast cancer cells and in tumor tissues. Western blotting analysis was applied to quantify the protein expression of HSP70 in the MCF-7 cells. Immunohistochemical staining was done to assess the positive rate of HSP70 expression in the tumor tissues.

RESULTSLeptin activated HSP70 in a dose-dependent manner in vitro: leptin upregulated significantly the expression of HSP70 at mRNA and protein levels in MCF-7 human breast cancer cells (P < 0.001). There was no significant difference in expression of HSP70 mRNA in the implanted tumors between the leptin-treated group and the control group (P > 0.05). Immunohistochemical staining revealed no significant difference in tumor HSP70 expression between the leptin-treated group and the control group (P > 0.05).

CONCLUSIONSA nude mouse xenograft model can be safely and efficiently treated with human leptin by subcutaneous injections around the tumor. HSP70 may be target of leptin in breast cancer. Leptin can significantly upregulate the expression of HSP70 in a dose-dependent manner in vitro.

Animals ; Blotting, Western ; Breast Neoplasms ; drug therapy ; metabolism ; Cell Line, Tumor ; Female ; HSP70 Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Immunohistochemistry ; Leptin ; pharmacology ; therapeutic use ; Mice ; Mice, Nude ; Real-Time Polymerase Chain Reaction ; Xenograft Model Antitumor Assays