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BACKGROUND:Polysulfone membrane holds good anti-biodegradation ability, but how to use it to prepare hol ow fiber dialysis membrane and its blood compatibility have not been ful y understood. OBJECTIVE:To study the preparation, transfer property and biocompatibility of hol ow fiber dialysis membrane. METHODS:With polysulfone as the film material, diethylene glycol as the porogen, polyvinyl pyrrolidone as the modifier, N, N-dimethylacetamide as the solvent, and the hol ow fiber dialysis membrane was prepared using nonsolvent-induced phase separation. The performance was measured using scanning electron microscopy, ultra-depth three-dimensional microscope imaging and porosity test;the transfer parameters including reject rate and water flux were detected by ultrafiltration device;the blood compatibility was determined through hemolysis test, dynamic clotting time test and platelet adhesion test. Type II medical polyurethane material served as negative control. RESULTS AND CONCLUSION:The section of hol ow fiber dialysis membrane was asymmetric. 17%dialysis membrane showed a porous middle layer, while 19%, 21%and 23%membrane showed a sponge-like middle layer. Under the same membrane area, the density of fiber dialysis membrane was significantly lower than that of the negative control material, and the porosity of fiber dialysis membrane was significantly higher than that of the negative control material (P<0.05). The water volume and water flux of the hol ow fiber dialysis membrane were significantly higher than those of the negative control material (P<0.05). Results from three hemolytic tests showed that the average absorbance values and hemolysis rate of the hol ow fiber dialysis membrane were significantly higher than those of the negative control material (P<0.05). The dynamic clotting time test and the platelet adhesion test revealed that the dynamic clotting time of hol ow fiber dialysis membrane at 20, 40 and 70 minutes was significantly shorter than that of the negative control material (P<0.05). These results suggest that polysiloxane can be used as the membrane material to prepare hol ow fiber dialysis membrane using nonsolvent-induced phase separation, and holds a good biocompatibility, blood compatibility and transfer efficiency.
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ObjectiveTo investigate the effects of over expression of miR-34a on cellular proliferation and migration in bladder cancer cell line J82 by targeting Notchl.MethodsmiR-34a was predicted as a putative gene which can target Notchl through bioinformatics analysis,qRT-PCR and Western blot were performed to measure the expression levels of Notchl and miR-34a in invasive transitional cell carcinoma of bladder (TCCB) tissues and J82 cells transfected with miR-34a.Luciferase assay was employed to determine if miR-34a could target Notchl through binding to the 3'-untranslated region (3'UTR) of Notchl mRNA.J82 cells were transfected with pcDNA3.0-miR-34a or pcDNA3.0 control plasmid.MTS colorimetry was used to evaluate the effect of miR-34a on cell proliferation.The effect of miR-34a on cell migration was assessed by transwell migration assay.ResultsThe expression level of miR-34 in invasive TCCB tissues was lower than in adjacent bladder tissues (0.016(0.018) vs 0.042 (0.059),N =16; P =0.0006).On the contrary,the average levels of Notchl mRNA and protein were higher in tumors than in adjacent bladder tissues (2.765(2.156) vs 2.312(1.365),N =16; P =0.0025 and 0.857 ±0.197 vs 0.648 ±0.171 ;P ＜0.0001 ).After the transfection of miR-34a,the expressive level of miR-34a in J82 was highly induced ( (2.408 ±0.789) × 10-4 vs(0.153 ±0.029) × 10-4; P =0.0026).However,the expressive levels of Notchl mRNA and protein were obviously decreased (3.001 ± 0.106 vs 4.998 ± 1.053 ; P =0.0308 and 0.747 ± 0.050 vs 0.988 ± 0.102 ; P =0.0215 ).The results of luciferase assay showed that firefly activity was highly dimished (0.422 ± 0.028 vs 2.392 ± 0.148 ; P ＜ 0.0001 ).Cellular proliferation was inhibited after the transfection of miR-34a in J82 (P ＜ 0.0001 ).Moreover,number of migration cells of J82 was significantly reduced after the ectopic expression of miR-34a ( 179.3 ± 21.02 vs 269.7 ± 23.71 ; P =0.0078 ).ConclusionsmiR-34a inhibits the cellular proliferation and migration of bladder cancer cell line J82 via binding to the 3UTR of Notchl mRNA.
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Objective To investigate the feasibility and safety of retroperitoneal laparoendoscopic single-site (LESS) donor nephrectomy using home-made single-port device.Methods From January 2011 to June 2012,11 consecutive LESS left donor nephrectomies using home-made single-port device with conventional laparoscopic instrument were performed through retroperitoneal access in our center.Results The procedures were completed and no complications occurred in all donors.Mean operative time was 149.5 min.Estimated blood loss was 30-350 ml.Warm ischemia time was 2-4 min.The urine output was prompt in all cases.Recipient graft function was normal within 2 weeks.Donor hospital stay was 5-6 days after operation.Conclusion LESS donor nephrectomy using home-made single-port device in our initial experience is feasible and safe.It is also cost-effective and minimally invasive with conventional laparoscopic donor nephrectomy.This technique is a good option for living donor nephrectomy.
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Retroperitoneal laparoscopic live donor nephrectomy offers an intrinsic advantage over conventional transperitoneal laparoscopic nephrectomy because of the potentially lower risk for early and late donor intraperitoneal complications. Herein we presented our experience performing retroperitoneal laparoscopic live donor nephrectomy in 105 donors. All donor nephrectomy was successful. There were no donor deaths and no conversion to open surgery. Mean operation time was 112 min (range, 70-200 min). Intraoperative blood loss was 10-150 mL with an average of 30 mL. Warm ischemia time was 1.3 to 6 min with an average of 3.1 min. Postoperative retroperitoneal hematoma occurred in only one case and there were no other surgical complications. Donors were discharged from the hospital 5 to 10 days postoperation. Average postoperative hospital stay was 6.4 days. One graft was removed due to acute rejection. Delayed graft function occurred in two recipients but renal function returned to normal within four weeks. The other recipients had normal renal function in two weeks except three recipients in four weeks. We believe that retroperitoneal laparoscopic live donor nephrectomy is safe, reliable, and less invasive.
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Objective To explore the effect of L-carnitine on panel reactive antibody (PRA) in hemodialysis patients. Methods 50 patients were classified randomly into 2 groups: L-carnitine group receiving intravenous injection of 2g L-carnitine after each hemodialysis for 6 months, and control group did not receive any L-carnitine treatment. The PRA in serum was measured by enzyme-linked immunosorbent assay (ELISA) before and after 6 months of L-carnitine treatment. Results L-carnitine significantly reduced PRA levels compared with control group(P
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Objective: To evaluate the effects of antisense oligodeoxynucleotides (ODNs) (AS1 complementary to the translation initiation region and AS2 complementary to the coding region) targeted to bcl-2 oncogene on Bcl-2 protein expression and apoptosis of human renal cell carcinoma (RCC) cells. Methods; Expression of bcl-2 mRNA in RCC cell lines was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR). The ODNs were transfected with Lipo-fectin into RCC cell lines. The expression of Bcl-2 protein in ACHN tumor cells was examined by Western blot analysis, and the apoptosis of those cells was determined by flow cytometric analysis. Results: Expression of bcl-2 mRNA was detected in all five RCC lines. Transfected bcl-2 antisense ODNs, but not sense ODNs, inhibited Bcl-2 protein expression in ACHN cells. The AS2 antisense ODN showed a superior effect compared with AS1 ODN. The apoptosis of ACHN cell could been induced by bcl-2 antisese ODNs , and percentage of apoptotic cells was noted 32. 1% and 43. 2% treated with AS1 and AS2, respectively. Conclusions: Treatment of human RCC cells with antisense ODNs targeting bcl-2 gene inhibits expression of Bcl-2 protein and induce apoptosis.
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Objective To construct a fusion minigene expression vector of seven HCV NS3 epitopes and express the fusion gene in the eukaryotic cell line, in order to form a foundation for the investigation of the DNA immunization for HCV prevention. Methods Three pairs of complementary oligonucleotides primers covering all seven HCV NS3 epitopes were synthesized. Overlapping extension PCR were performed to construct the fusion minigene and cloned in pEGFP-N3 and pBuDCE vector respectively. The fusion protein was detected by Western blotting and flow cytometric analysis. The transfection efficiency was assessed with by flow cytometric analysis. Results The HCV NS3 epitope fusion minigene was obtained and inserted into pEGFP-N3 and pBuDCE vector respectively. The recombinant plasmid pEGFP-DR4 and pBuDCE-DR4 were expressed in 293T and 046W cell lines respectively. The Western blot result indicated that the expected 36 kDa minigene/EGFP fusion product was expressed from pEGFP-DR4, while 13kDa product from pBuDCE-DR4 respectively. Conclusion The HCV NS3 epitope fusion minigene was expressed in the eukaryocyte expression system.