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.

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.

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 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

OBJECTIVE: To investigate the prevalence rate, malocclusion type and treatment rate as well as awareness of malocclusion among children and adolescents living in the Hangzhou municipality. METHODS: 1818 children and adolescents ages 7 approximate, equals 16 living in Hangzhou municipality were evaluated. RESULTS: The overall malocclusion rate was 35.75%. In Angle's classification: Angle I 593 cases (32.62%), Angle II 48 cases(2.64%),Angle III 9 cases (0.49%). Among the malocclusion type of crowding was mixed dentition 70.66%,permanent dentition 85.89%. In the overjet malocclusion mixed dentition was noted in 60.57% and permanent dentition in 51.05%.The type of overbite was mixed dentition 67.82%,permanent dentition 31.23%. In the crossbite of anterior teeth mixed dention was noted in 12.30% and permanent dention in 9.91%. Overall treatment rate for malocclusion was 10.15%.CONCLUSION: Among Hangzhou municipality juveniles there is both inadequate prevention and treatment of dental malocclusion.

The aim of this study was to investigate the tissue factor (TF) expression of platelets and leukocytes in patients with acute coronary syndrome (ACS), patients with stable angina (SA) and healthy subjects (as controls). 26 patients with ACS, 29 patients with SA, and 25 controls were enrolled in this study. The peripheral blood samples of above-mentioned subjects were collected and isolated to obtain the monocytes and platelet-rich plasma, the TF-mRNA expression of monocytes, and platelets among 3 groups was detected by RT-PCR, the TF expression ratio of platelets, platelet-leukocyte aggregates (PLA) and platelet-monocyte aggregates (PMP) was detected by flow cytometry among 3 groups. The results showed that the TF mRNA expression level of platelets in ACS group were significantly higher (3.11 ± 0.51 relative expression) as compared with SA and control groups (1.88 ± 0.78 and 0.7 ± 0.1, respectively) (P = 0.03). Expression of TF mRNA of monocytes was higher in ACS group (P = 0.05 versus controls) too. ACS group had a significantly higher amount of TF-positive platelets (8.8 ± 2.6) than SA (2.6 ± 0.5, P = 0.02) or control groups (2.5 ± 0.4, P = 0.02). A significantly greater number of TF positive platelet-leukocyte aggregates and platelet-monocyte aggregates were also found by flow cytometry in blood of ACS patients than in either SA patients or controls. It is concluded that the high TF expression of platelets and leukocytes in ACS patients strengthens the platelet activation, blood coagulation, and thrombus formation and may further contribute to the hypercoagulability associated with the disease. The present study further extends the proinflammatory/prothrombotic phenotype of ACS patients showing that new players on the scene.
Acute Coronary Syndrome ; blood ; Aged ; Angina, Stable ; blood ; Blood Platelets ; metabolism ; Case-Control Studies ; Cell Adhesion ; Female ; Humans ; Leukocytes ; metabolism ; Male ; Middle Aged ; Platelet Aggregation ; RNA, Messenger ; metabolism ; Thromboplastin ; metabolism