Objective: Our aim was to study the concentrations and regulatory characteristics of heparin-binding epidermal growth factor-like factor (HB-EGF) and its functional consequences in inducing the activities of hexokinases (HKs) and net Glc utilization. Methods: The SV40MES13 cell line was used as cell model for this experiment to measure the total activity of Hks by using a standard G6PDH-coupled assay. The net Glc utilization was measured indirectly as Glc disappearance from the medium using the standard Glc oxidase-coupled reaction(Trinder). Results: The HB-EGF induced HKs activity in time- and dose-dependent manners and increased the net Glc utilization by SV40MES13 cells. Conclusion: The HB-EGF plays an important role in pathogenesis of diabetic nephropathy by inducing total HKs activity specifically and increases the net Glc utilization in mesangial cells.

Objective To provide guide for clinical diagnosis,classification and treatment of chronic kidney disease-mineral and bone disorder(CKD-MBD).Methods A total of 56 patients with chronic renal insufficiency were divided into three different groups:CKD4 period,CKD5 period and long-term hemodialysis(over 5 years).Their serum calcium,phosphorus,iPTH,bAP,and TRACP levels,and the bone mineral density were examined.Results The incidences of CKD-MBD in the three groups were 36.4%,73.0%,and 87.5%,respectively.High-turnover bone disease accounted for 65.79%,and the low-turnover bone disease accounted for 34.21%.The bAP and TRACP activity was significantly increased in patients with high-turnover bone disease and decreased in those with low-turnover bone disease.Conclusion CKD-MBD is a common complication of patients with chronic renal failure,with the main type being high-turnover bone disease,but the low-turnover bone disease can not be ignored.TRACP and bAP are the sensitive indicators of osteoblasts and osteoclast activity,and they could be used to identify high or low turnover bone disease.TRACP and bAP,together with serum calcium,phosphorus,iPTH,bone mineral density changes,can be used for a comprehensive analysis of bone metabolic state,so as to guide the diagnosis,sub-typing,drug selection and comprehensive treatment of CKD-MBD.

Objective:To explore the relationship between Feng syndrome(风证) and plasma endothelin(ET)in initial condition of acutely ischemic stroke.Methods:Using the table of syndrome quantitative diagnosis which formulated according to experiences of apoplectic experts to grade the fraction of syndrome which appears in initial condition of 151 cases with acute cerebral infarction，at the same time the level of plasma ET was determined by radioimmunoassay(RIA),and then according to the fraction of syndrome patients were divided into two groups,one is Feng syndrome group,the other is nonFeng syndrome group，the levels of plasma ET in this two groups and with healthy subject were compared.Results:It showed that the level of plasma ET in Feng group was significantly higher than that in nonFeng group（P<0.001）；and the same result between the patients and the healthy subjects（all P<0.001） were obtained.Conclusions:The results demonstrate that there is a correlated relationship between Feng syndrome and plasma ET,and the level of plasma ET can be regarded as a microcosmic indices to separate Feng syndrome from nonFeng syndrome.

Objective To investigate clinical effect of sEMG-BFB training combined with swallowing training for dysphagic patients with cerebral infarction,to provide a reference for life quality improvement of dysphagic patients.Methods 48 patients with cerebral infarction combined swallowing disorder were selected.According to the digital table,patients were divided into the control group and observation group,24 cases in each group.Patients in the control group were given training such as routine training and behavioral swallowing training,while patients in the observation group were given sEMG-BFB training based on training of the control group.FOIS score,Kubota's water drinking test grade,SSA score and SWAL-QOL score were compared between two groups.Results After treatment,FOIS score,SSA score and SWAL-QOL score of the observation group were (3.82 ± 1.04),(19.97 ± 8.17),(158.23 ± 10.27),which were significantly higher than those of the control group [(2.82 ± 1.21),(24.21 ± 6.31),(139.23±7.32),t =4.394,4.115,5.642,all P ＜ 0.05] ; Kubota's water drinking test grade of the observation group was better than the control group,the difference was statistically significant (U =283.43,P ＜ 0.05).Conclusion sEMG-BFB training combined with swallowing training is helpful for dysphagic patients with cerebral infarction to improve swallowing ability and life quality.

Objective To observe the effect of MG132 on the expression of extracellular regulated kinase 1/2 (ERK1/2) and connective tissue growth factor (CTGF) in rat peritoneal mesothelial cells (RPMCs) induced by high glucose.Methods RPMCs were isolated,cultured and passaged by trypsin,then identified.The second generation of cultured RPMCs were used in the experiment.RPMCs were divided into normal control group,high glucose (1.5％,2.5％,4.25％) for 24 hours,high glucose (2.5％) for 0,12,24,48 hours,incubated with MG132 (0.5,1,2 μmol/L) for half an hour and then with high glucose (2.5％) for 24 hours.ERK1/2 protein was detected by Western blotting,and CTGF protein in supernatant was detected by ELISA.Results Compared with the control group,the expression of p-ERK1/2 was significantly increased in the groups stimulated by high glucose (P ＜0.01),reached the peak at 24th hour (P ＜ 0.01),and then the expression decreased at 48th hour,but still was higher than that in the normal control group (P ＜ 0.01).CTGF protein expression of RPMCs induced by high glucose increased,in time-and dose-dependent manner (P ＜ 0.05).MG132 could significantly decrease the expression of ERK1/2 and CTGF induced by high glucose (P＜0.05).Conclusions MG132 can decrease the expression of p-ERK1/2 and CTGF in RPMCs induced by high glucose.The ubiquitin proteasome pathway participates in the development of peritoneal fibrosis,and blocking the way may contribute to the prevention of peritoneal fibrosis.

[Objective] To construct a database for the genetic polymorphism of 18 STR loci (D8S1179, D21S11, D7S820, CSFIPO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, VWA, TPOX, D18S51, D5S818, FGA, PentaE, PentaD, SE33) in Hart population from Zhejiang province. To investigate the application of 18 STR loci in the field of paternity testing and prenatal diagnosis. [Methods] Fluorescent dye labeling multiplex STR-PCR, capillary electrophoresis and DNA sequencer GeneScan were adopted in genotyping 598 unrelated samples collected from Han population in Zhejiang province. 18-STR database was established and analyzed. Population comparison was conducted between Han population in Zhejiang province and 8 other population. 15-STR and 18-STR identification system were compared in 497 paternity testing cases. [Results] We observed the distribution of 18 STR loci in Han population meet Hardy-Weinberge equilibrium and was different from other 8 population (X~2 test, P>0.05). Statistical results showed that the heterozygosis (He) ranged from 0.630 to 0.942. The combined power of discrimination was>0.9999999999. Compared with 15-STR identification system, higher paternity index scores and higher exclusion rate were obtained with 18-STR identification system in dual-case paternity test and mutation identification. One trisomy 21 fetus was found in a prenatal paternity test case which had two characteristic genotypes in 2 STR loci of D21S11 and Penta D. [Conclusions] The 18 loci were relatively highly genetic polymorphic in Zhejiang Han population and could be used for paternity testing. Some STR loci could be used in prenatal diagnosis for aneuploidy.

Objective To observe the effect of Xuebijing injection on the expression of tumor necrosis factor-alpha (TNF-α) and high mobility group box-1 protein (HMGB-1) in rat peritoneal mesothelial cells (PMCs) induced by lipopolysaccharide (LPS). Methods PMCs were isolated from rat colic omentum and the 3rd generation cells were used in the experiment. PMCs were incubated with LPS at different concentrations (1,10,100 mg/L);with LPS (10 mg/L) for 2, 6, 12, 18, 21, 24, 36 h;with Xuebijing injection at different concentrations (2,10,20 g/L) after incubation with LPS (10 mg/L) for 2 h. PMCs in the control group were incubated with medium. HMGB-1 mRNA was detected by RT-PCR. TNF-α and HMGB-1 protein in supernatants was detected by ELISA. Results Compared to the control group, the expression of HMGB-1 mRNA and protein was significantly increased in groups stimulated by LPS in a time- and dose-dependent manner (all P<0.05);the expression of TNF-α was increased in the groups stimulated by LPS in a dose-dependent manner (P<0.05). In the groups stimulated by LPS (10 mg/L), the expression of TNF-α appeared double hump within 36 hours. Compared to LPS (10 mg/L) group, Xuebijing injection significantly inhibited the expression of HMGB-1 and TNF-α (all P<0.05 ) in a dose-dependent manner. Conclusions HMGB-1 as a late mediator of inflammatory responses may play a role in the pathogenesis of peritoneal dialysis related peritonitis. Xuebijing injection can reduce peritoneal inflammatory impairment by inhibiting the up-regulation of TNF-α and HMGB-1 induced by LPS.

Objective To observe the effect of ulinastatin on the expression of interleukin 15 (IL-15), connective tissue growth factor (CTGF) and malondialdehyde (MDA) in rat peritoneal mesothelial cells (RPMCs) induced by high glucose. Methods RPMCs were isolated, cultured and passaged by trypsin, then identified. The third generation of cultured RPMCs were used in the experiment. RPMCs were divided into normal control group, high glucose (1.5%, 2.5%, 4.25%) for 6 hours and 12 hours, high glucose (2.5%) for 3, 6, 12, 24 hours or ulinastatin (160, 320, 640U/ml) for 12 hours. IL-15 mRNA was detected by real-time PCR. IL-15 and CTGF protein in supernatants was detected by ELISA. MDA protein was detected by TBAS. Results Compared with the control group, the expression of IL-15, CTGF and MDA was significantly increased in the groups stimulated by high glucose (P＜0.05) in dose- and time-dependent manner. Ulinastatin could significantly decrease the expression of IL-15, CTGF and MDA induced by high glucose in dosedependent manner both in protein and gene levels (P＜0.05). Conclusions High glucose can up-regulate the expression of IL-15, CTGF and MDA in RPMCs. Ulinastatin can reverse these changes.

Objective To observe the effect of 1,25(OH)_2D_3 on expression of vitamin D receptor (VDR), tumor necrosis factor α (TNF-α) and transforming growth factor β1 (TGF-β1) in rat peritoneal mesothelial cells (RPMCs) stimulated by lipopolysaccharide (LPS), so as to provide some evidence for clinical use of 1,25 (OH)_2D_3 on peritoneal dialysis-associated peritoneal inflammation. Methods RPMCs were isolated, cultured and passaged by enzymaticdisaggregation, then identified by phase contrast inverted microscope, transmission electron microscope with immunocytochemistry method. RPMCs were incubated with LPS (1,10,100 mg/L) and LPS (10 mg/L) for 2,6,12 hours, or stimulated by 1,25(OH)_2D_3 (10~(-8) mol/L, 10~(-7) mol/L, 10~(-6) mol/L) after incubated with LPS (10 mg/L) for 2 hours. RPMCs in the control group were justincubated with medium. Expression of VDR mRNA and protein was detected by RT-PCR and Western blot. In addition, ELISA was performed to investigate the changes of TNF-α and TGF-β1 in culture medium. Results Compared with control group, the expression of VDR mRNA and protein was decreased in LPS group (P<0.05). LPS could significantly induce the expression of TNF-a and TGF-β1 in RPMC (P <0.01), which could be partially reversed by different concentrations of 1,25 (OH)_2D_3 (P<0.01). Conclusions 1,25 (OH)_2D_3 can reverse the decrease of VDR mRNA and protein induced by LPS as well as the induction of TNF-a and TGF-β1 up-regulated by LPS in RPMC in a dose- and time-dependent manner. Our research provides experimental evidence of VDR ameliorating peritoneal dialysis-associated peritoneal inflammation and peritoneal fibrosis.

Objective:To describe a feasible surgical technique for patients with renal cell carcinoma associated with a supradiaphragmatic tumor thrombus that avoids cardiopulmonary bypass procedure. Methods:We retrospectively analyzed 2 cases with right kidney tumor and tumor thrombus above the dia-phragm treated in April and August,2015.The two patients were both female,aged 73 and 67 years. The tumor sizes of right kidneys were 7.0 cm ×6.3 cm ×5.7 cm and 8.7 cm ×7.0 cm ×5.2 cm,and the tumor thrombuses were 1.3 cm and 1.8 cm above the diaphragm.The second patient had synchro-nous metastasis in right adrenal gland ,and the tumor thrombus arose from the adrenal vein but not the renal vein.Intraoperative transesophageal echocardiography (TEE)was used to assess real-time mobility of the thrombus.A modified chevron incision was used,the right kidney was mobilized laterally and pos-teriorly,and the renal artery was identified,ligated,and divided.The infradiaphragmatic inferior vena cava (IVC)was exposed and isolated by mobilizing the liver off the diaphragm or to the left (piggyback liver mobilization,case 2).The central diaphragm tendon was dissected or incised in the midline until the supradiaphragmatic intrapericardial IVC was identified and gently pulled beneath the diaphragm and into the abdomen.The tumor thrombus was then “milked”downward out of the intrapericardial IVC un-der the guidance of TEE.The distal and proximal IVC to the tumor thrombus,porta hepatis,and left re-nal vein were clamped.Tumor thrombus was removed from the IVC.The IVC was sutured and vascular clamps were placed below the major hepatic veins.Pringle’s maneuver was then released and hepatic blood drainage was permitted during closure of the remaining IVC.Related literature was reviewed.Re-sults:Complete resection was successful through the transabdominal approach without CBP in both pa-tients.Estimated blood loss was 1 500 mL and 2 000 mL,and 1 200 mL and 800 mL of blood were trans-fused.The postoperative courses were uneventful.Both patients subsequently underwent tyrosine-kinase inhibitor therapy.Both patients were alive without tumor recurrence or new metastasis during the follow-up of 6 months and 9 months.Conclusion:In selected cases,renal cell carcinoma extending into the IVC above the diaphragm can be resected without sternotomy,CBP or DHCA.