ObjectiveTo investigate the clinical curative effect of traditional Chinese medicineKuiyu decoction combined with quadruple therapy for peptic ulcer (PU).MethodsThis study was a randomized controlled trial. 82 patients with PU and conformed to the indusive criteria were randomly divided into a control group and a treatment group by the random number table method, with 41 cases in each group. The control group was treated with quadruple therapy, while the treatment group was treated with traditional Chinese medicineKuiyu decoction based on the treatment of the control group. Integral change of main symptoms before and after treatment and the level changes of serum motilin (MTL), somatostatin (SS) of the two groups before and after treatment were compared and clinical total effective rate was evaluated.ResultsThe total effective rate was 95.1% (39/41) in the treatment group and 80.5% (33/41) in the control group. There was statistical difference between the two groups(χ2=4.100,P=0.043). Stomach pain (0.58 ± 0.21vs.0.73 ± 0.28,t=2.744), gasteremphraxis (0.35 ± 0.12vs.0.43 ± 0.14,t=2.778), sour regurgitation integral (0.28 ± 0.09vs.0.34 ± 0.11,t=2.703) and serum MTL level (358.5 ± 81.3 pg/mlvs.422.8 ± 86.9 pg/ml,t=3.460) in the treatment group were lower than those in the control group (P<0.01);the serum SS level (87.5 ± 18.7 pg/mlvs.73.8 ± 17.9 pg/ml,t=3.389) in the treatment group was higher than that in the control group(P<0.01).ConclusionsTraditional Chinese medicineKuiyu decoction combined with quadruple therapy for the patients with PU can promote SS secretion through decreasing MTL, so it can improve the clinical effect.

Objective To study the effect and mechanism of electroaeupuncture at acupoint Guanyuan on expression of brain-derived neurotrophic factor (BDNF) and tropomyosine receptor kinase B (TrkB) in spinal cord and excitability of detrusor of rat with urinary retention after spinal cord injury. Methods The model of urinary retention after spinal cord injury was established by using weight drop method. Eighty female Sprague-Dawley rats were randomly divided into four groups, namely, a normal, a sham, a model and an electroacupuncture (EA) group, with 20 rats in each group. The EA group was given electroacupuncture at aeupoint Guanyuau once a day for 10 days in total, while the other groups was given no treatment. After 10 days, the expression level of BDNF and TrkB in spinal cord of injury site was measured by immunohistochemistry staining, and excitability of detrusor was measured by using the muscle strips in vitro. Results It was revealed that, with regard to the expression level of BDNF and TrkB in spinal cord,the model group exceeded the normal and sham group, the EA group significantly exceeded the other groups ( P < 0. 05 ). With regard to the excitability of detrusor, the model group was significant-ly lower than the normal and sham group. The EA group significantly exceeded the model group ( P < 0.05 ). Conclusion Electroacupuncture at acupoint Guanyuan can raise the expression level of BDNF and TrkB in spinal cord and increase the excitability of detrusor in rats with urinary retention after spinal cord injury.

Objective To assess the effect of a noninvasive equipment (the human biological stimulation feedback instrument) for the screening of metabolic syndrome (MS). Methods The samples were selected from the International Health Care Center of the Second Affiliated Hospital of Zhejiang University College of Medicine. From January to December 2012, according to the inclusion criteria, a total of 5674 consecutive participants (3 437 men, 2 237 women) without medical history of diabetes mellitus underwent general health screening in this hospital, which included questionnaire investigation, all MS components and the risk assessment of the human biological stimulation feedback instrument. MS was defined according to 2004 the Chinese Diabetes Society (CDS) criteria. Results The 5 674 subjects were divided into three groups in accordance with the human biological stimulation feedback instrument risk score: the normal group, metabolic abnormalities low risk group, metabolic abnormalities high risk group. The detection rate of MS corresponding to the three groups was 8.73% (251/2 876), 29.32% (612/2 087), 32.77% (233/711), respectively (trend χ2=372.14,P<0.001). With the increase of risk score, the detection rate of MS increased. A total of 1 096 (male 879, female 217) were diagnosed MS by CDS criteria. The area under the curve (AUC) of the test was 0.674 (95%CI 0.658-0.691, P<0.000 1)for predicting MS. The corresponding cutoff value was 25.00%, the sensitivity, specificity, positive predictive value and negative predictive value of diagnosis of MS was 77.80%, 56.00%, 30.20%and 91.30%, respectively;409 subjects who had no history of MS and diabetes mellitus follow up for two years, the AUC of the test was 0.719 (95%CI 0.644-0.794)for predicting MS. The corresponding cutoff value was 25.00%, too. And the sensitivity and specificity of diagnosis of MS was 78.00%and 59.80%, respectively. The detection rate of MS was positively correlated with the risk score. Conclusion We found that the human biological stimulation feedback instrument is a potential screening approach for early predicting MS in health checkup with simple and noninvasive procedure and high negative predictive value, although its diagnostic efficiency is not so good. And it might have a warning effect on the progress of MS.

Objective To investigate the effects of the overexpression of HO-1 induced by CoPP in the donor on the survival of transplanted allogeneic islets of rats and the mechanism.Methods (1) Brown Norway rats were randomly divided into control group, and CoPP-induced group receiving intraperitoneal injection of CoPP (2.5 mg/kg) at 3rd and 1st day prior to islet isolation.By using the cytoimmunofluorescenee and Western blot, the expression of HO-1 in isolated islets was detected.The insulin level in the supernatant of the cultured islets stimulated with glucose was determined by ELISA.(2) Lewis male rat diabetic models were established by a single intravenous injection of alloxan, and then randomly divided into CoPP group and control group.Islets were transplanted under the left kidney capsule of each diabetic recipient.The survival time after transplantation, and pathological changes following rejection of the islet grafts were analyzed.Results The HO-1 was highly expressed in the islets isolated from CoPP-treated rats by cytoimmunofluorescence and Western blot.After stimulation with 16.7 mmol/L glucose, the insulin concentration in Copp-treated and Copp-untreated groups was (46.60± 1.13) and (19.01 ± 1.49) mIU/L respectively (P<0.05).The insulin concentration in Copp-treated and Copp-untreated groups in islets stimulated with 5.6 mmol/L glucose was (15.65 ± 0.89) and (12.28 ± 0.89) mU/L respectively (P>0.05).The stimulated index in Copp-treated and Copp-untreated groups was (2.98 ± 0.10) and 1.55 + 0.01 respectively (P< 0.05).The survival time of islets allograft in Copp-treated and Copp-untreated groups was separately (12.20±5.67) and (5.60± 1.14) days respectively (P<0.05).Histological analysis revealed the presence of more islands of insulin-positive cells and considerably fewer lymphocytes or inflammatory infiltration than the controls.Conclusions CoPP could induce the HO-1 expression of islets, and improve their function.Over-expression of HO-1 in islets could prolong survival time of islets allograft.

Objective To study the effect of electro-acupuncture(EA)on bladder detrusor apoptosis in rats with urinary retention after spinal cord injury(SCI),compare the effectiveness of different acupoints and explore possible mechanisms.Methods One hundred female adult Sprague-Dawley rats were randomly divided into 5 groups:a normal group,a sham operation group,a model control group,a Guanyuan acupoint(RN4)EA group and a Shuidao acupoint(ST28)EA group,with 20 rats in each group.A model of urinary retention after SCI was established using the weight drop method.The EA groups were given EA once daily for 10 d.The other groups had no treatment.After 10 d of EA,an electron microscope was used to observe the bladder detrusor ultrastructure,and the terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL) method and immunohistochemistry were used to observe any apoptosis in the detrusor smooth muscle cells and the expression of apoptosis-related genes bcl-2 and bax.Results After EA treatment,expansion of the rough endoplasmic reticulum was reduced significantly,and the number of edemic mitochondria was reduced compared with the model control group.In the normal and sham operation groups,little apoptosis was observed,and there was no significant difference between the two groups.Apoptosis indices(AIs)were higher in the model control group than in the normal group.The average AI in the Guanyuan EA group was lower than that in the Shuidao group.In the Guanyuan group,bax protein expression was less than that in the Shuidao group and the model group.In the Guanyuan group,bcl-2 protein expression was not significantly different from that in the Shuidao group and the model control group.Conclusion EA at the Guanyuan or Shuidao aeupoint can restrain bladder detrusor apoptosis in rats with urinary retention after SCI.EA may improve the ultrastructure of the detrosor urinae, reduce the expression of bax, change the ratio of bcl-2 to bax, and thus decrease apoptosis.The effect of EA at the Guanyuan acupoint is superior to that at the Shuidao acupoint.

Objective To detect the expression levels of interleukin (IL)-25 and IL-33 in peripheral blood of patients with chronic urticaria.Methods Ninety-three patients with chronic urticaria were included in this study along with 30 healthy individuals.All the patients were treated with loratadine for four weeks.Blood samples were collected from the healthy controls and patients before and after the four-week treatment.Enzyme-linked immunosorbent assay (ELISA) was performed to detect the serum levels of IL-25 and IL-33.The relationship between the expression levels of the two cytokines and urticaria severity was analyzed.Results The serum levels of IL-25 and IL-33 in the patients before treatment were significantly higher than those in the healthy controls (IL-25,139.86 ± 28.48 vs.114.41 ± 34.00 ng/L,P ＜ 0.01; IL-33,91.95 ± 21.88 vs.79.80 ± 30.72 ng/L,P ＜ 0.05),and positively correlated with the severity of urticaria (r =0.38,0.42,respectively,both P ＜ 0.01).After four weeks of treatment,clinical improvement was observed in 81.72％ of these patients with a significant decrease in the serum IL-25 level (116.48 ± 21.94 vs.139.86 ± 28.48 ng/L,P ＜ 0.01),but no obvious changes in the serum IL-33 level (90.88 ± 20.62 vs.91.95 ± 21.88 ng/L,P ＞ 0.05) compared with those before treatment.Conclusions The expressions of IL-25 and IL-33 are elevated in peripheral blood of patients with chronic urticaria,and positively correlated with the severity of urticaria.

Objective To establish the method for isolation and culture of porcine Sertoli cell sand detect the expression of the immune privilege-related molecules in the cultured cells. MethodsTestes were aseptically removed from the 10-to 15-days old Large White piglets. Testes were decap-sulated, minced and then digested with 0.2% (W/V) collagenase type V, 0.25% (W/V) trypsin and 0.05% (W/V) DNaseI. In the primary isolation, Sertoli cells were cultured at 37℃with 5% CO2.Inverted phase contrast microscopy and HE staining were used to observe the morphology of Sertoliceils, and the Sertoli cells were identified under an electron microscope. Viability and apoptosis ofcultured cells were measured with the AnnexinV-PI staining by flow cytometry. The expression of sox9, FasL, TGF-β and clusterin in Sertoli cells was detected by RT-PCR. The viability of long-termcultured Sertoli cells was assayed by MTT. Results In the cultured total cells, Sertoli cells accountedfor more than 90%. The apoptosis rate and mortality of Sertoli cells was (2.61±0.96)% and (2.12±0.74)% respectively. RT-PCR revealed that the expression of sox9, TGF-β and clusterin in theSertoli cells was strongly positive, but FasL was weakly positive. Viability of cultured cells measuredby MTT conformed that the sertloli cells could survive more than 21 days. Conclusion The isolationand culture methods of the neonatal porcine Sertoli cells was established. Under the culturedconditions, the Sertoli cells can express the immune privilege molecules and successfully survive at least 21 days.

Objective To investigate whether cotransplant with xenogenetic neonatal porcine Sertoli cells (NPSCs) could prolong rat islet allograft survival and its mechanisms.Methods 1500 islets equivalent quantity (IEQ) and 1×10~7 NPSCs were implanted under renal capsule of diabetic Wistar rats.Islets implanted alone were used as control group (n=6);islets co-transplanted with NPSCs under left renal capsule of recipients served as experimental group (n=6);meanwhile,islets and NPSCs implanted into the different sides of kidneys were used as another control grouP(n=4).Blood glucose level was measured everyday.The graft-bearing kidneys at the time of rejection were Results Co-transplantation with NPSCs to the same site significantly prolonged islet allograft survival (mean survive time,16.3±1.4 days vs.5.7±1.0 days in islet transplant alone control group,P<0.05).In contrast,transplantation with NPSCs and islets separately did not prolong the islet allograft survival (5.3±0.5 days).HE staining showed plenty of local infiltrated lymphocytes in the transplanted site of the eontrol group.which were demonstrated as mainly CD3+ T cells by immunopathology.The local expression of Bcl-2 was markedly elevated in co-transplantation group as compared with the other 2 groups,while there were no significant differences in the HO-1 expression among these groups.Conclusion Co-transplantation with xenogenic NPSCs can significantly prolong islet allograft survival in rats.The immunoprotective mechanism may be associateel with the inhibition of lymphocyte infiltration and the enhancement of the local expression of protective gene Bcl-2.

Objective To investigate whether porcine Sertoli cells eould resist xenoreactive antibodies mediated complement lysis. Methods Sertoli cells were isolated from testes of 10 to 15 day-old landrace pigs. Α-Gal expression on Sertoli cells was measured by FACS and cytoimmunofluorescence. The binding of human se-rum IgG and IgM with Sertoli cells was assayed by FACS. After the incubation of the cultured Sertoli cells with 20% human B serum in vitro, the cellular lysis and cytotoxicity assay were detected with CytoTox-ONETM homogeneous membrane integrity assay and MTT method, and then activation of the complement cascade was examined by immunohistochemistry and immunofluorescence. The SV40-porcine endothelium cells line (SV40-PED) was served as control cells. Results α-Gal expression was found on Sertoli cells by FACS and cytoimmunofluorescence. After incubation with 20% human serum, cellular lysis ratio of the SV40-PED was 53. 13% ± 14.53%, while lysis ratio of the Sertoli cells was significantly lower (24.38% ±0.50%, P<0.01), the viability of Sertoli cells and SV40-PED were 98.73% ± 18.84% and 52.43% ± 8.08%, respectively. With immunohistochemistry and immunofluorescence, C3c and C4d were found binding on both the Sertoli cells and SV40-PED cells, however, C5b-9 was only detected on SV40-PED cells. Conclusion In vitro, compared with the SV40-PED, Sertoli cells could resist xenoreactive antibodies of human serum mediated complement lysis by preventing the C5b-9 formation.

ObjectiveTo explore the expression of N-cadherin and β-catenin mRNA in human brainstem and supratentorial gliomas. MethodsN-cadherin and β-catenin mRNA expression in 18 cases of brainstem gliomas and 18 cases of supratentorial gliomas tissues were detected with PT-PCR. Resultsβ-catenin mRNA expression was more in human brainstem gliomas than in supratentorial gliomas （t=2.255,P<0.05）, but was not significantly different of N-cadherin mRNA (P＞0.05). The expression of N-cadherin mRNA in human brainstem gliomas of grades Ⅰ～Ⅱ were less than those in human gliomas of grades Ⅲ～Ⅳ (t=2.711，P<0.05), but was not of β-catenin mRNA (P＞0.05). N-cadherin mRNA expression was positively correlated with the β-catenin mRNA expression in either brainstem gliomas or supratentorial gliomas （r=0.480，r=0.809 respectively, P<0.05）. ConclusionThe over expressions of N-cadherin and β-catenin may play an important role in the invasion and malignant progress of human brainstem gliomas.