GDF-15,a distant member of the TGF-superfamily,is identified as an apoptotic accelerat- ing,anti-tumorigenesis and nerve2nutritional factor in varied injures and tumors and has cardioprotective ac- tivity.The characteristics and roles of GDF-15 gene/protein and antibodies are expounded besides the rela- tionship between GDF-15 serum level/genetypes and CRC.It is also discussed here that some antitumorigenic substances inducing GDF-15 in CRC tissues and CRC cells.

Objective: To construct a polycistron tandem repeated Echistatin (Ecs) gene. Methods: Three Ecs genes with independent initiation and termination codon were ligated tandem through restriction enzyme sites after amplified with 3 pairs of primers using pMD18T-Ecs as template. The polycistron Ecs gene was inserted into pET30a and expressed in E.coli BL21(DE3) with IPTG induction. The expression results were identified by 18% SDS-PAGE and Western blot. Results: The expression of Ecs polycistron was accomplished with 18% expression level of total protein determined by SDS-PAGE and Western blot. Conclusion: The successful expression of Ecs polycistron provided a new method for the preparation of low molecular weight protein.

Humanin(HN,its analogue [Gly14]-Humanin,HNG)was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults.But the relative low content of this peptide in its natural sources limits its further characterization.An expression vector pET32a/HNG was corstructed and transformed it into E.coli BL21 trxB(DE3).HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography.Subsequently,the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC.A 23 mg recombinant HNG(rHNG)from 1 L bacterial culture was purified.The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide.The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence.Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

Humanin (HN, its analogue [Gly14]-Humanin, HNG) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults. But the relative low content of this peptide in its natural sources limits its further characterization. An expression vector pET32a/HNG was corstructed and transformed it into E. coli BL21 trxB (DE3). HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography. Subsequently, the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC. A 23 mg recombinant HNG (rHNG) from 1 L bacterial culture was purified. The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide. The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence. Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

Kidney is a highly vascularized organ and peritubular capillary network constitutes the critical component of its microvascular system. Peritubular capillaries, as the main vessels for blood supply in renal tubules and renal interstitium, involve in important physiological processes in renal tubules such as energy metabolism, substance secretion and reabsorption. In recent years, it has been demonstrated that ischemia-reperfusion injury, rejection and renal fibrosis during kidney transplantation would result in compromised structural integrity and decreased number in peritubular capillaries, thus leading to interstitial fibrosis in renal allograft, which would seriously affect the long-term stability of renal function in the renal allograft. Therefore, we reviewed the structure and function of peritubular capillary, peritubular capillary and ischemia-reperfusion injury, rejection and renal allograft fibrosis, focusing on the mechanism for peritubular capillary injury in kidney transplantation and the specific changes manifested, with the aim of providing a reference for preventing and treating perioperative complications in kidney transplantation and improving the long-term prognosis of grafts.

Objective To evaluate tension-free hernia Onlay repair with premuscular positioning of the prosthesis for the treatment of ineisional henia of the abdominal wall.Methods In this study 126 patients with incisional henia were treated with a tension-free manner of hernia repair by using synthetic material ONLAY between September 1999 and June 2007.Results All operations were successful.There was no hospital death or severe postoperative complications.The average age was 58.5 years old ranging from 28 to 89.There were 67 patients in which the abdominal defect ranged from 5～10 cm,and 59 patients with abdominal defect≥10 cm.The mean operating time was 95(70～120)min,and the average intraoperative blood loss was 80 ml(60～250 ml).The mean postoperative hospitalization was 14.5 days(10～28 d). Patients were followed-up from 3 to 96 months,and 3 patients suffered from hernia recurrence(2.38%). Conclusions The ONLAY repair of ineisional hernia of the abdominal wall with synthetic material mesh was a safe procedure,especially for those with large abdominal wall defects.

OBJECTIVETo assess the efficacy and safety of percutaneous cryoablation (PCC) and (125)I seed implantation combined with chemotherapy for advanced pancreatic cancer.

METHODSSixty-seven patients with advanced pancreatic cancer (6 in stage III, 61 in stage IV) received PCC and (125)I seed implantation combined with concomitant gemcitabine hydrochloride and DDP chemotherapy. The clinical benefit response (CBR), survival rate and therapy-related complications were assessed.

RESULTSAll patients except one were followed up over 1 year. The 6-month and 1-year survival rates were 84.8% and 33.4%, respectively. The median progression free survival were 6.3 months and 5.5 months in the group stage III and group stage IV (P > 0.05), respectively, while the overall survival was 9.1 months in the group stage III and 11.0 months in the group stage IV (P > 0.05). CR,PR and SD were achieved in 5, 8, 54 patients, respectively. Fifty-four and 50 in the 67 patients experienced a ≥ 50% reduction of pain score and analgesic consumption, respectively, 18 patients experienced a ≥ 2 kg weight gaining, and KPS was increasing from 71.2 ± 0.4 to (90.0 ± 0.3, P < 0.05), the overall benefit rate was 80.6%. No serious therapy-related complications except pancreatic fistula accompanied abdominal hemorrhage, bile leakage, acute pancreatitis and needle track seeding in 1, 1, 2 and 1 case, respectively.

CONCLUSIONPercutaneous cryoablation and (125)I seed implantation combined with chemotherapy are effective and safe for the treatment of advanced pancreatic cancer.

Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Brachytherapy ; methods ; Cisplatin ; administration & dosage ; Combined Modality Therapy ; Cryosurgery ; methods ; Deoxycytidine ; administration & dosage ; analogs & derivatives ; Disease-Free Survival ; Female ; Follow-Up Studies ; Humans ; Iodine Radioisotopes ; therapeutic use ; Liver Neoplasms ; secondary ; Male ; Middle Aged ; Neoplasm Staging ; Pancreatic Neoplasms ; pathology ; surgery ; therapy ; Survival Rate ; Young Adult

OBJECTIVETo study whether necroptosis exists or not in neural cell death induced by aluminum.

METHODSSH-SY5Y cells were treated with 4 mmol/L AlCl(3) x 6H(2)O The cell viability was determined with CCK-8 kit after treated with Nec-1 at different dosages (0, 30, 60, 90 micromol/L). Mitochondria membrane potential (MMP), content of reactive oxygen species (ROS), and apoptotic rate/necrotic rates were measured with cytometry.

RESULTSNec-1 ameliorated the necrotic-like cell morphology, the cell viability were 0.28 +/- 0.05, 0.58 +/- 0.03, 0.68 +/- 0.04, and 1.03 +/- 0.17, there were significant differences between the Nec-1 treated groups and that of controls (t values were 3.25, 3.36, 4.56; P < 0.05). After Nec-1 treatment, the necrotic rates were 16.46% +/- 0.54%, 10.40% +/- 0.64%, 5.43% +/- 0.68%, and 6.28% +/- 0.35%, there were significant differences between the Nec-1 treated cells and that of controls (t values were 3.62, 7.32, 6.96; P < 0.05); while the apoptotic rates were 8.68 +/- 0.36, 7.66 +/- 0.53, 5.68 +/- 0.41, and 4.13 +/- 0.41, there was no significant difference among the groups (F = 6.33, P = 0.11). Cytometry had shown the increased cell MMPs after Nec-1 treatment, which were 67.54 +/- 6.36, 49.42 +/- 5.96, 84.79 +/- 6.86, and 95.51 +/- 7.01, there were significant differences as comparing MMPs of the middle and high dosage of Nec-1 treated cells with those of controls (t values were 3.21, 4.01; P < 0.05); while ROS contents in the Nec-1 treated SH-SY5Y cells were 54.07 +/- 3.32, 52.79 +/- 2.36, 54.68 +/- 1.91, and 59.23 +/- 2.96, there was no significant difference among the groups (F = 5.26, P = 0.19).

CONCLUSIONNec-1, as a specific inhibitor of necroptosis, might effectively block the cell death pathway induced by aluminum, it indicates that necroptosis should be one of the major causes of the SH-SY5Y cell toxicity induced by aluminum, and necroptosis also plays an important role in aluminum induced SH-SY5Y cell death.

Aluminum ; toxicity ; Apoptosis ; drug effects ; Cell Death ; drug effects ; Cell Line, Tumor ; Humans ; Imidazoles ; pharmacology ; Indoles ; pharmacology ; Neuroblastoma

OBJECTIVETo study the effect of necrostatin (Nec-1) on apoptosis induced by aluminum (Al), and approach the mechanism.

METHODSNeural cell death model was made by 4 mmol/L Al treated neuroblastoma cells (SH-SY5Y). Cell viabilities were detected at different concentrations of Al and/or Nec-1. Hoechst 33342/PI double staining was used to observe apoptosis and (or) necrosis that were quantified by flow cytometry using Annexin V/PI double staining. Apoptotic pathway was tested by activities of Caspase-3, Caspase-8 and Caspase-9. In addition, the expression of NF-kappa B and Cyt-c was measured by immunocytochemistry.

RESULTSCell viabilities were significantly decreased with the increasing concentrations of Al (P < 0.05), which could be significantly upregulated by 60 micromol/L Nec-1 (P < 0.05) and were correlated with the concentrations of Nec-1 (P < 0.05, P < 0.01). Apoptosis and necrosis were observed under fluorescent microscope and quantified by flow cytometry, which suggested an increasing trend of apoptotic and necrotic rates (P < 0.05, P < 0.01). Whereas, Nec-1 could not only decrease the necrotic rate but also apoptotic rate as well (P < 0.05, P < 0.01). Data of Nec-1 on caspases activities showed that Nec-1 could not affect Caspase-9 activity (P > 0.05) and Cty-c protein expression as well (P > 0.05). However, Nec-1 could reduce Caspase-8 activity significantly (P < 0.05, P < 0.01) and increase NF-kappa B protein expression (P < 0.05, P < 0.01) and finally decrease Caspase-3 activity (P < 0.05).

CONCLUSIONNec-1 could reduce cell apoptosis induced by Al, through Caspase-8 pathway, and up-regulate the expression of NF-kappa B protein.

Aluminum ; toxicity ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Death ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Cytochromes c ; metabolism ; Humans ; Imidazoles ; pharmacology ; Indoles ; pharmacology ; NF-kappa B ; metabolism ; Neuroblastoma

OBJECTIVEThe purpose of this study was to classify the diffuse large B-cell lymphoma (DLBCL) into different prognostic subgroups according to four different detection methods of the expression of CD138, CD10, bcl-6, and MUM1. In particular to investigate the significance of CD138 in immunohistochemical profiles and its correlation with prognosis in DLBCL.

METHODSImmunohistochemical EnVision method was used to detect the expression of CD138, CD10, bcl-6 and MUM1 in 106 cases of DLBCL and reconstructed into four different subtyping algorithms. Algorithm-1, according to the expression of CD10, bcl-6 and MUM1, the cases were assigned to GCB and non-GCB groups. Algorithm-2, according to the expression of CD138, CD10, bcl-6 and MUM1, the cases were assigned to A, B, C, D groups. Algorithm-3, according to the expression of CD10 and MUM1, the cases were assigned to GCB and non-GCB groups. Algorithm-4, according to the expression of CD138, CD10, bcl-6 and MUM1, the cases were assigned to GCB and non-GCB groups. Following up was included as well. Statistical analysis was performed using the SPSS 13.0 and differences were considered significant at P < 0.05.

RESULTSCD138, MUM1, CD10 and bcl-6 were positive in 15.1% (16/106), 56.6% (60/106), 21.7 (23/106) and 26.4% (28/106), respectively. The expression of CD10 and bcl-6 was associated with favorable OS (P = 0.001 and 0.041, respectively), whereas the expression of CD138 was associated with unfavorable OS (P = 0.003). Using multivariate Cox proportional hazards regression analysis, algorithm-1 and -4 were almost at the same level for prognosis of OS (OR = 0.259, 0.255) and PFS (OR = 0.248, 0.244).

CONCLUSIONSBoth Hans's algorithm and Colombo's algorithm including CD138 detection are associated with the prognosis of DLBCL patients. The two algorithms have similar OR value according to Cox analysis. However, positive expression of CD138 is of minor significance in prediction of the prognosis in DLBCL patients.

Humans ; Immunohistochemistry ; Lymphoma, B-Cell ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; Prognosis ; Syndecan-1 ; metabolism