Objective Radiosensitivity is highly correlated with DNA doub le stra nd breaks(DSBs). The repair of human DSB is possible mainly through nonhomologou s DNA end joining(NHEJ)pathway. This study was designed to determine the relat ionship between expression of 70Ku/ 80Ku prot ein (a modulating subunit of DNA-PK, which is an important component in NHEJ pathway) and radiosensitivity of nasoph aryngeal carcinoma cell lines. Methods Radiosensitivity parameters of the nasoph aryngeal carcinoma cell lines were obtained by the colony forming assay and radi ation dose-survival curve was done by linear quadratic model. Western blot was p erformed to determine the expression of 70Ku and 80 Ku in total extract of CNE1 a nd CNE2 at baseline level、different time after 4?Gy irradiation、12 h after di ff erent doses of irradiation. To perform a semi-quantitated assay of protein expr e ssion, the optic density (OD) value of each band was estimated by automatic imag e analysis system. Results Surviving fraction of CNE1 was higher than those of CNE2 at each dose point. Mean inactivation dose was higher in the CNE1(2.35)t han that in the CNE2(1.11), but the quantity of 70Ku/ 80Ku in either cell line was similar at both baseline and postirradiation levels. Conclusions CNE2 is mo re radiaosensitive than CNE1; as there was no marked correlation observed betwee n the quantity of Ku protein and radiosensitivity of the nasopharyngeal carcinom a cell lines. X-irradiation cannot result in any change in total quantity of Ku protein.

Objective To compare the sensitivity of HE,immunohistochemistry (IHC) and RT-PCR in detection of breast cancer metastases in axillary lymph nodes.Methods Twenty female patients with newly diagnosed and clinically node-negative breast cancers underwent modified radical mastectomy, including a complete axillary lymph node dissection. The ages of the patients ranged from 31 years to 65 years, and the diagnosis of breast cancer was approved by pathological finding. Two hundred and thirty-nine axillary lymph nodes were found in these 20 patients. Metastases in axillary lymph nodes were explored by HE, cytokeratin 19 IHC and RT-PCR for cytokeratin 19 respectively. Results Seven(2.9%) lymph nodes were found to have metastatic cancers by HE in 3 patients,all nodes were found in level Ⅰ. Metastatic cancers were found in 13 (5.4%) nodes by IHC in 7 patients,11 nodes in level Ⅰ and 2 nodes in level Ⅱ; and 52(21.8%) nodes were found to contain tumor cells by RT-PCR in 14 patients,30 nodes in level Ⅰ and 22 nodes in level Ⅱ. All of 7 histologically(HE) positive nodes were found to contain tumor cells by IHC and RT-PCR. Among 232 histologically(HE) negative nodes,6(2.6%) nodes were found to contain tumor cells by IHC,and 45(19.4%) nodes were found to contain tumor cells by RT-PCR, all 6 IHC positive nodes showed the expected 460-base pair products on gel electrophoresis ( P

Objective Partial hepatectomy, liver transplantation, and radio frequency ablation for hepatocellular carcinoma (HCC) were compared to select the most suitable method for HCC. Methods 1198 patients with HCC in 3 hospitals in Guangzhou were divided into 3 groups: group Ⅰ , small HCC; group Ⅱ > HCC without vascular invasions and group Ⅲ , HCC with vascular invasion. The patients either received partial hepatectomy, transplantation or ablation. The 1-, 2- or 3-year survival rates, the 3-year recurrent rates and Child-Pugh grades in the 3 groups were compared. Results For small HCC, there was a significant increase in the 3-year survival rate (P<0. 05) and a significant decease in the recurrent rate (P<0. 05) in patients who received transplantation, compared with those who received hepatic resection. Patients who received ablation had a higher 3-year survival rate and a lower recurrence (P<0. 05) in comparison with those who received hepatectomy. There was no significant difference(P<0. 05) between transplantation and ablation, but there were more Child A patients who received hepatectomy and ablation, and more Child B and C patients who received transplantation. For advanced HCC, there was no significant different in the 3-year survival rates for the 3 therapies, but the 3-year recurrence was lower (P<0. 05) in the transplantation group. Conclusions For small HCC, superiority of transplantation versus resection was obvious. Ablation (diameter <3 cm) was also superior to resection, whereas ablation was as effective as transplantation. There were more Child B and C patients in the transplantation group than the ablation and resection groups. Therefore, small HCC with hepatic decompensation should receive liver transplantation. Transplantation was advantageous in having less tumor recurrent but there was no difference in the 3 therapies for advanced HCC.

Objective To study the pathogenesis of upper gastrointestinal rehaemorrhagia after the transjugular intrahepatic portasystemic shunt (TIPS) and its influencing factor.Methods Fifty postoperative patients with TIPS were selected.The patients were followed-up,and the effect of the various factors in the role of upper gastrointestinal rehaemorrhagia after TIPS was analyzed.Results The portal vein pressure of 50 patients with TIPS decreased from preoperative (39.8 ±9.2) cmH2O (1 cmH2O =0.098 kPa) to postoperative (25.2 ± 5.8) cmH2O,and there was statistical difference (P ＜ 0.05).Fourteen patients appeared upper gastrointestinal rehaemorrhagia after TIPS,which accounted for total of 28％ (14/50) and included 3 cases of postoperative vomiting blood within 3 days.Acute stomach mucosa lesions bleeding was considered,and bleeding was controlled within a short-term medical treatment (1 patient after more than a year in recurrent upper gastrointestinal rehaemorrhagia).Twelve cases of patients appeared upper gastrointestinal rehaemorrhagia within 2 years after TIPS,and the causes of rehaemorrhagia in 6 cases were esophageal variceal rehaemorrhagia,gastric and duodenal ulcer in 3 cases,erosive gastritis in 2 cases,coagulation abnormalities in 1 case.Esophageal variceal rehaemorrhagia rate was 12％ (6/50).Conclusions The main reasons of upper gastrointestinal rehaemorrhagia after TIPS are variceal rehaemorrhagia and non variceal rehaemorrhagia,both of which are important causes of rehaemorrhagia after TIPS.Variceal rehaemorrhagia after TIPS occurs more than 3 months,and non variceal rehaemorrhagia occurs within 3months,so it is very important to protect gastric mucosa with proton pump inhibitor in postoperative patients.

Objective To set up the molecular cytobiological model of endogenous coagulation factor Ⅷ (FⅧ) re-expressing in human liver cells L02,and to study the regulation pathway and molecular basis of the re-expression of FⅧ in L02 cells activated by NO signal.Methods The L02 cells at logarithm growth phase were selected and randomly divided into blank control group and experimental group, inhibitor group and inhibitor control group;they were cultured for 0,12,24,36,48,and 60 h.Flow cytometry was used to detect the expression of human FⅧ protein in L02 cells after treated for 48 h.Griess experiment was performed to detect the levels of NO in L02 cells at different time points;the transcription levels of human FⅧ gene,iNOS gene,NF-κB1 gene and I-κB alpha gene were detected by RT-PCR method.Western blotting method was used to detect the expression levels of human phosphorylated I-kappaB (phosphorylated I-κB)in L02 cells.Results The results of flow cytometry showed that the expression of human L02 FⅧ protein was found after treated with L-arginine for 48 h. The Griess results showed that the levels of NO in L02 cells in experimental group were significantly increased at 3,6,12,and 24 h (P<0.05)and the levels of NO in blank control group,inhibitor group and inhibitor control group had no changes. The RT-PCR results showed that the transcription of human FⅧ mRNA in L02 cells was found in experimental group,but there was no transcription of human FⅧ mRNA in blank control group,inhibitor group and inhibitor control group;the transcription levels of iNOS,NF-κB1 and I-κB alphain experiment group were increased(P<0.05)and the transcription levels of these genes in blank control group,inhibitor group and inhibitor control group had no changes. The Western blotting results showed that after adding L-arginine the expression level of phosphorylated I-κB was significantly increased (P < 0.05 ), other groups had no such change. Conclusion L-arginine can activate the phosphorylation of I-κB by NO signal pathway to lead to the changes in the expression of human FⅧ gene promoter upstream regulatory-related transcription factors NF-κB to activate the expression of human FⅧ in human liver cells L02.

Diterpenes, an important class of natural compounds, are widely distributed in nature. As the valuable diterpenoids continue to be found, diterpene synthase in the course of diterpene synthesis get as much attention as possible. The multiformity of end-product-diterpenoids were also due to the diversity of diterpene synthase. This paper focuses on the advances in recent biosynthesis pathway of diterpene and types, cloning, catalytic mechanism, synthetic biology application.

BACKGROUND: Previous studies have shown that a variety of materials can be used for the construction of tissue engineering scaffolds. The topological structure of the scaffold surface has a regulatory effect on the biological behaviors such as stem cell proliferation and differentiation, but the specific mechanism is still unclear. OBJECTIVE: To investigate the role of P38 and Akt pathways in the oriented differentiation of bone marrow mesenchymal stem cells in nanofiber scaffolds. METHODS: Three kinds of nanofiber scaffolds (AFS, AYS, 3-DPS) with different structures were constructed. Rat bone marrow mesenchymal stem cells were inoculated on the surface of three kinds of nanofiber scaffolds. After osteogenic induction, cell morphology, adhesion and proliferation were detected. mRNA expression levels of key phenotype molecules (COLIα1, COLIIα1, Aggrecan, Sox-9) were measured using qRT-PCR. Intracellular P38, AKT, ERK1/2 and JNK expression was detected by western blot assay. RESULTS AND CONCLUSION: After 4 and 8 hours of culture, cell adhesion rate of the 13-DPS scaffold group was higher than that of the AFS and AYS scaffold groups (P<0.05). After 7 days of culture, cells of the 13-DPS scaffold group proliferated faster than those of AFS and AYS scaffold groups (P<0.05). Bone marrow mesenchymal stem cells adhered firmly and grew well on three kinds of scaffolds. Fibroblast-like growth was observed on the AFS and AYS scaffolds and chondrocyte-like growth was observed on the 3-DPS scaffold. After 3 weeks of cartilage induction, mRNA expression of COLIIα1, Aggrecan and Sox-9 was higher, and the mRNA expression of COLIα1 was lower, in the 3-DPS scaffold group compared with the other two groups (both P<0.05). After 3 weeks of cartilage induction, relative expression level of p-AKT and p-P38 in the 3-DPS scaffold group was significantly higher than that in the other two groups (both P<0.05). There were no significant differences in AKT total protein and ERK1/2, JNK, P38, p-ERK1/2, p-JNK and p-P38 protein expression levels among three groups. These findings suggest that nanofiber annulus fibrosus scaffolds with different spatial structures can induce the oriented differentiation of bone marrow mesenchymal stem cells through the P38 and AKT pathway, which were the downstream of the Integrin-FAK signaling pathway.

Objective To demonstrate the imaging characteristics of pancreatic neuroendocrine neoplasm (P-NEN).Methods The CT and MRI findings of 43 cases of P-NEN confirmed by pathology were analyzed,and compared with pathological results.Results Mean size of 43 lesions in 43 P-NEN patients were (25.00± 1.82)mm.Necrosis cystic degeneration were found in 26 lesions.In CT plain scan,15 lesions were isodense,28 lesions were hypodense.All of 43 lesions were hypointensity signal on T1WI;27 lesions were hypointensity signal,16 cases were isointensity signal on T2WI.Twenty-seven lesions showed envelope;30 lesions showed clear outline.In enhancement images,24 lesions were marked enhancement since arterial phase;8 lesions showed delayed enhancement;5 lesions showed centrality enhancement;6 lesions showed wild enhancement.Conclusion P-NEN has some characteristics on CT and MRI scanning,imaging can play an important role in diagnosis for P-NEN.

Objective:To investigate cytokine(TNF-?,IL-1) effect on apoptosis and express of related protein in thyrocytes and the relationship with pathogenesis in Graves' disease(GD).Methods:To test thyroid tissues from 50 patients with GD Fas expression by immunohistochemical method.Thyroid tissues were collected from samples of GD operation and were cultured by primary culture method.Content of sFas in culture medium was tested by ELISA.Fas/sFas mRNA were tested by RT-PCR.Cytokine inducing apoptosis in Graves' disease showes observation group,normal people showes control group.Results:Compare apoptosis culture rate there is significant difference(P

Objective To compare the advantages and disadvantages of smear immunohistochemistry and biopsy of bone marrow in the detection of non-Hodgkin lymphoma (NHL) infiltration. Methods 60 newly diagnosed patients with NHL were collected. Smear immunohistochemistry and biopsy of bone marrow were applied to detect bone marrow involvement in those patients. The relations between the results of two detection methods and the relevant clinical characteristics, such as age, blood routine examination, lactic dehydrogenase, extensive infiltration, B symptom, performance status, clinical stage and so on were analyzed. Results Positive rate of bone marrow involvement in bone marrow smear immunohistochemistry group was higher than that in bone marrow biopsy group [10.0 % (6/60) vs. 3.3 % (2/60), P=0.008]. When detection on B cell infiltration, positive rates were 6.6 % (4/60) and 3.3 % (2/60) in smear immunohisto chemistry group and biopsy group, respectively (P=0.007), and when detection on T cell infiltration, the positive rates were 3.3%(2/60) and 0 (0/60), respectively. Furthermore, two detection method was not correlated with related clinical characteristics, such as bone marrow biopsy, immunohistochemistry, gender, age, Karnofsky score, B symptom, extranodal involvement, lactic dehydrogenase, thrombocyte, Hb, neutrophils, lymphocyte and staging (all P>0.05). Conclusions The positive rate of bone marrow smear immunohistochemistry is superior to bone marrow biopsy in the detection of bone marrow involvement of NHL. As for the immunophenotyping of NHL, sensitive rate of bone marrow smear immunohistochemistry is also better than that of bone marrow biopsy.