目的：分析斑秃患者外周血白细胞介素‐17（IL‐17）和 T淋巴细胞亚群的变化及其意义，以探讨该病的发病机制，为临床治疗方案的制订提供依据。方法选取2014年7月至2015年5月该院确诊的斑秃患者86例，另选取40例健康者作为对照组，采用双抗体夹心法和流式细胞学方法检测外周血IL‐17和T淋巴细胞亚群，并进行比较分析。结果局限性斑秃组CD3＋T淋巴细胞与CD4＋ T淋巴细胞百分比均低于对照组，差异有统计学意义（ P＜0．05）；普秃、全秃组CD3＋ T淋巴细胞、CD4＋ T淋巴细胞、CD8＋ T淋巴细胞百分比均低于对照组，差异有统计学意义（ P＜0．05）；普秃、全秃组CD3＋ T 淋巴细胞与CD8＋ T淋巴细胞百分比均低于局限性斑秃组，差异有统计学意义（ P＜0．05）；对照组40例（100．00％）IL‐17百分含量均小于1％，斑秃组仅44例（51．16％）小于1％，IL‐17百分含量为1％～＜3％、3％～＜5％、≥5％的患者分别占18．60％、12．79％、17．44％，明显高于对照组，差异均有统计学意义（ P＜0．05）。结论斑秃患者反映体内总淋巴水平的CD3＋ T 淋巴细胞低于健康人群，局限性斑秃以CD4＋ T淋巴细胞为主，全秃、普秃由CD4＋ T淋巴细胞和CD8＋ T淋巴细胞共同作用所致。

Introduction and Aims: Although it is reported that gut microbiota and colon barrier function were deteriorated in CKD state, the detailed mechanism or pathological relevance has not been elucidated. Oral activated charcoal adsorbent (AST-120) has been reported to delay the progression of CKD by adsorbing uremic toxin from the intestine. However, the effects of AST on the gut environments in CKD and its impact on CKD progression have not been fully elucidated. Methods: Six-week-old spontaneously hypertensive rats (SHR) were rendered CKD by 5/6 nephrectomy (Nx). The SHRs were divided into four groups; group 1, sham-operated SHR (SHR, n=10); group 2, SHR given AST-120 (SHR+AST, n=10); group 3, 5/6 nephrectomized SHR (Nx, n=10); group 4, 5/6 nephrectomized SHR given AST-120 (Nx+AST, n=10). AST-120 was orally given at a dose of 4g/kg/day by mixing with regular chow. After 12 weeks, rats were sacrificed and biochemical parameters, histological changes in the kidney and colon, and molecular changes were compared among the groups. The distribution of the intestinal flora was examined by T-RFLP (Terminal Restriction Fragment Length Polymorphism) and real time PCR. Human colon cell line, Caco-2 cell were treated with uremic toxin precursor, indole. Results: Serum level of indoxylsulfate (IS), one of the major uremic toxin was increased in Nx, which was significantly reduced in Nx+AST. Serum creatinine levels and urinary protein excretion were increased in Nx, which were attenuated in Nx+AST. Glomerular sclerosis and tissue fibrosis evaluated by Masson-trichome staining were increased in Nx, which were also attenuated in Nx+AST. The histological analysis of the colon tissue revealed that the number of goblet cell and the protein expression of mucin-2 were decreased in Nx. These decreases were restored by AST. Although mRNA levels were not altered, the expression levels of tight junction protein ZO-1 and Claudin-1 were decreased in Nx, which were mitigated in Nx+AST. The analysis of the gut flora showed the decrease of the Lactobacillus in Nx. This decrease was also reversed in Nx+AST. Regression analysis revealed that the number of Lactobacillus in colon was significantly associated with serum creatinine levels. In Caco-2 cells, treatment with indole, precursor of IS produced in the colon downregulated mucin-2 expression as well as Occludin and ZO-1 expressions. Conclusions: In the previous reports, tight junction proteins were downregulated and intestinal barrier function was impaired in mucin-2 deficient mice. It has been also reported that mucin-2 enhanced the growth of Lactobacillus by inducing their adhesion to the colon. Our data demonstrated that AST improved the gut environment favorable to Lactobacillus which affected the expressions of the tight junction proteins. And indole directly influences the tight junction and mucin-2. These effects provide a novel mechanism whereby AST120 or probiotic therapy improves renal function through modulating the gut environment.

Objective:To assess the clinical efficacy and the impact of astragalus particles on T lympho-cyte subsets in peripheral blood among the patients with alopecia areata. Methods:A total of 113 patients with AA were recruited and divided into treatment group ( group A) and control group ( group B) . Oral compound glycyrrhizin tablets, 50 mg/time and 3 times a day, was prescribed to the patients in group B and oral com-pound glycyrrhizin tablets in a combination astragalus granule, 4 g/time and 2 times a day were given to the patients in group A. The treatment course was 3 months for both groups. The T lymphocyte subsets in peripher-al blood was detected by flow cytometry. Results: The recovery rate and the recurrence rate of the treatment group were superior to the control group (P<0.01 and P <0.01). CD4+T cells increased significantly in the treatment group, compared with the results before treatment and the control group after treatment. The differ-ences were statistically significant ( P<0.05 and P<0.01) . Conclusion:Astragalus granule combined with om-pound gycyrrhizin is effective in the treatment of alopecia areata.

OBJECTIVE:To observe the effects of ryanodine on rapamycin treated endothelial outgrowth cells (EOCs). METHODS:The mononuclear cells were harvested from umbilical cord blood by Ficoll density gradient centrifugation, then induced into EOCs and expanded in vitro. The endothelial characteristics of EOCs were identified by immunostaining and fluorescent staining. The EOCs were pretreated with or without ryanodine (10 µmol/L) for 1 h, and then treated with or without rapamycin (10 nmol/L) for 24 h. Proliferation was evaluated by CCK8 and migration was measured by Transwell. The protein expression of EOCs was evaluated by immunobloting technique with total eNOS antibody and phospho-eNOS (Thr495) antibody. RESULTS:Compared with control group, the proliferation and migration capacities of EOCs were significantly reduced while the phosphorylation of eNOS (Thr495) protein was significantly upregulated in rapamycin group (P < 0.05), expression of total eNOS was not affected by rapamycin (P > 0.05). Compared with rapamycin group, the proliferation and migration capacities of EOCs were significantly increased and the phosphorylation of eNOS (Thr495) protein was significantly downregulated in ryanodine + rapamycin group (P < 0.05). The proliferation and migration capacities, the phosphorylation of eNOS (Thr495) protein and the expression of total eNOS were not affected by ryanodine alone (P > 0.05). CONCLUSIONS:Rapamycin reduced proliferation and migration capacities while upregulated the phosphorylation of eNOS (Thr495) protein of EOCs and these effects could be partly reversed by cotreatment with ryanodine.

目的 探讨慢性阻塞性肺疾病急性加重(AECOPD)患者多耐药鲍曼不动杆菌(MRAB)感染及与营养状况的相关分析.方法 采取2008-01 -2011-02 AECOPD患者的痰标本,对64株鲍曼不动杆菌进行分析.结果 药敏测定提示亚胺培南对该菌敏感率最高,为81.3％.其中18例为MRAB感染,其中中低蛋白血症12例,低体重指数14例.结论 鲍曼不动杆菌是AECOPD常见的感染细菌,MRAB感染已经占一定比例.营养状况较差的COPD患者易发生MRAB感染.

子宫肌瘤是女性生殖器官最常见的良性肿瘤,其发生率为20%～30%[1].近年来,子宫的生理功能和机体器官的完整性受到更多的关注,使子宫肌瘤剔除术的应用日益增加,尤其是随着微创技术的发展,临床上开始施行腹腔镜下子宫肌瘤剔除术(laparoscopicmyomectomy.LM).但到目前为止,对LM的推广仍有争议.现收集我们两家医院妇产科近年来行LM患者的资料进行分析,并与同期开腹行子宫肌瘤剔除术(abdominalmyomectomy,AM)的92例患者作一对照,旨在探讨LM应用的安全性、可行性及临床应用价值.

Objectives: To evaluate the effect of medical interventions on the sonographic appearance of endometrium in the infertile women with endometrial abnormalities and the clinical outcome of controlled ovarian stimulation cycles.Methods: Transvaginal ultrasonography was performed in follicular phase in natural period.30 of 60 infertile patients found with abnormal endometrial echoes were treated with hysteroscopy assisted dilation curettage(HADC group),another 30 with dilation curettage(DC group).The controls were 50 infertile women without identifiable endometrial abnormalities.The controlled ovarian hyperstimulation cycles were performed after treatment.Results: In HADC group and in DC group,the rate of type A endometrial echoes was significantly increased in controlled ovarian hyperstimulation cycles than in nature cycles,and the rate of type B,type C echoes was significantly decreased(P0.05).But compared with that in control group,the rate of type A endometrial echoes was significantly lower in HADC group and in DC group,and the rate of type B,type C echoes was significantly higher(P0.05).The clinic pregnancy rate in HADC group(56.7%) was statistically higher than that in DC group(43.3%) and in control group(40.0%)(P0.05).There was no significance difference between DC group and control group(P0.05). Conclusion: It is suggested that dilation curettage,especially hysteroscopy assisted dilation curettage,may be benefit to improve intrauterine disorder and increase clinic pregnancy rate.

Objective: To explore the value and moleculor mechanism of local injury therapy in improving the endometrial receptivity. Methods: There were 120 infertile patients who had 140 IVF cycles, in which 60 of them were in the local injury group (group A) and the other 60 women were in control group (group B). Scanning electron microscopy and immunohistochemistry were used to screen for the expression of pinopode, LIF and ostepontin (OPN) in the endometrium of two groups. Twenty non-pregnant women (group B2) from group B were taken to undergo local injury therapy and proceed with IVF treatment. The influence of the expression of pinopode, LIF and osteopontin before and after local injury therapy were also compared. Results: The expression of implantation related factors in group A and group B of the IVF patients were following: pinopode abundance 63.33% vs 36.67%, LIF 6.07 ± 1.33 vs 4.94 ± 2.26, OPN 6.36 ± 1.49 vs 5.28 ± 2.06. They showed more expression of pinopode and higher level of LIF, OPN in the endometrium of group A than in group B (P<0.05). The single embryo implantaion rate in group A was 27.78%, which was much higher than that in group B (14.29%, P<0.05). The expression of pinopode, LIF and OPN in group B2 before and after local injury treatment were following: pinopode abundance 10% vs 50%,LIF 4.50 ± 0.82 vs 6.33 ± 1.25, OPN 5.06 ± 0.98 vs 6.83 ± 1.46, respectively (P<0.05). A total of 140 IVF cycles were divided into the pregnancy group (62 IVF cycles) and the non-pregnancy group (78 IVF cycles) according to the outcomes of IVF-ET/ICSI. The implantation related factors expressed in the pregnancy group and the non-pregnancy group were following: pinopode abundance 54.84% vs 25.64%, LIF 6.94 ±1.03 vs 4.24±1.84, OPN 7.31±0.85 vs 4.52±1.65, respectively. These were also higher expression of the implantation related factors in the pregnancy group than in the non-pregnancy group (P<0.05). And they were correlative each other. Conclusion: Pinopode, LIF and OPN are important related factors in the implantation window. The abundant expression of pinopode and higher level of LIF, OPN in the endometrium may be closely related to the endometrial receptivity to implantation. Further more, the local injury may increase these implantation related factors and thus inhence the rate of IVF pregnancy.

Objectives: To evaluate the efficacy and hysteroscopic findings in patients undergoing IVF-ET who failed to conceive. Methods: Thirty-eight cases who had previous failed IVF-ET (experimental group) and 45 cases with infertility (control group) were analyzed. Intrauterine lesions on hysteroscopy were compared. Results: The incidence of abnormal endometrial findings in experimental group (84.21%) was significantly higher than that in control group (48.89%)(P<0.05). Intrauterine adhesion (36.84% vs. 20.00%), irregular endometrial hyperplasia (28.95% vs. 15.56%), and endometrial polyps (13.16% vs. 6.67%) were also significantly higher in experimental group than in control group (P<0.05). Thirty-two in 38 patients underwent following IVF/ICSI cycle within one year, 17 cases achieved pregnancy, clinical pregnancy rate was 53.13%. Conclusion: Hysteroscopy is highly valuable and should be routinely performed to all infertility cases before IVF, especialy with in vitro fertilization and embryo transfer failure. The morphological characters of endometrium directly affected the outcome of IVF.

Objective To discuss the effect of spontaneous falling of serum estradiol on outcomes in in vitro fertilizationembryo transfer(IFV-ET) cycles. Methods In 19 patients with spontaneous falling of estradiol levels in IVF-ET cycles,clinical manifestations,sex hormone levels and embryo characters were compared with the control group. Results Estradiol falling group was associated with fewer oocytes retrieved,lower estradiol level on the day of human chorionic gonadotropin administration,on-going pregnancy rate and baby birth rate compared to the controls. Conclusion The spontaneous falling of serum estradiol in IVF-ET cycles has adverse effect on IVF outcomes.However,the mechanism needs further study.

Objective: To assess the clinical value of a single maternal serum β-human chorionic gonadotropin(β-HCG) assay 11 days after embryo transfer in COS(Controlled Ovarian Stimulated) cycle and predict the following pregnancy outcome. Methods: A total of 192 pregnancies following embryo transfer were included. Invalid pregnancies were defined as biochemical pregnancies, ectopic pregnancies and the first trimester abortions. Ongoing pregnancies were defined as singleton pregnancies, twins pregnancies and multiple pregnancies whose gestation were achieved more than 12 weeks. Serum β-HCG concentrations were compared among different groups, and their relationship with pregnancy outcomes were analyzed. Results: At the post-embryo-transfer day 11, the mean β-HCG concentration of the ongoing pregnancy group (323.75±315.29miu/ml) was significantly higher than that of the invalid pregnancy group (81.43±73.15miu/ml) (P<0.001). In multiple gestations, the levels of β-HCG were significantly higher compared with singleton pregnancies(P< 0.001). If the β-HCG level was 10-50U/L, the positive predictive value of biochemical pregnancies and ectopic pregnancies was 81.8% , the negative predictive value was 94.4%. If β-HCG level was ≤100U/L, the positive predictive value of the first trimester abortions was 80.8%, the negative predictive value was 77.8%. If β-HCG level was﹥250U/L, the positive predictive value of multiple pregnancy was 83.3%, the negative predictive value was 74.4%. Conclusions: A single serum β-HCG level at the day 11 after embryo transfer is an effective predictive value for clinical pregnancy outcome in COS(Controlled Ovarian Stimulated) cycle and helps plan the subsequent follow-up.

Objective To study the effective methods of improving the pregnancy rate in IVF-ET. Methods From May 2003 to June 2005, practice embryo transfer guided by ultrasound、blood E- 2 /P monitoring to regulate the timing of HCG injection and endometrium activation before IVF cycle respectively among patients undergoing IVF-ET in Reproductive Center of Ruijin Hospital Affiliated to Shanghai Jiaotong University, who had fixed other treatment procedure. Results We found that on the day of ET, nearly 45.69% patients uterus position changed and 23.28% patients uterus depth increased or decreased more than 5mm, implantation rate of those ET under B-ultrasound was higher than others. After choosing the timing of HCG injection by regulating blood E- 2 /P, patients proportion of E- 2 /P ratio between 3.01 to 4.00 increased obviously and the proportion of less than 1.0 decreased. Through endometrium activation, 63.64% patients' endometrium abnormal previously were improved and these patients got the same pregnancy rate as those having normal endometrium. By exercising above strategies, our pregnancy and implantation rate had been increasing every year and reached 53.85% and 31.88% respectively. Conclusion ET under B-ultrasound and endometrium activation before IVF cycle shows their effect in improving the pregnancy rate of IVF-ET. While the definition of the value of monitoring blood E- 2 /P to regulate the timing of HCG injection needs more samples.

To investigate the function of ALK3 gene, the gene regulation and the signaling pathway related to ventricular septum defect during heart development. The model mice with ALK3 gene knock-out via alpha-MHC-Cre/lox P system were bred. The mRNA expression level of control group was compared with that of experiment group and ALK3 downstream genes were screened using PCR-select cDNA subtraction microarray. The mRNA of control group was extracted from E11.5 normal mouse hearts, and that of experiment group, from E11.5 hearts of mice with alpha-MHC Cre(+/-) ALK3(F/+) genotype. It was found that the mice with ALK3 gene knock-out produced heart defects involving the interventricular septum. The platelet-activating factors acetylhydrolase and the transcription factor Pax-8 and so on, were down-regulated. However, the Protein Tyrosine Kinase (PTK) of Focal Adhesion Kinase (FAK) subfamily and beta subtype protein 14-3-3 were up-regulated in the alpha-MHC Cre(+/-) ALK3(F/-) mice. These data provide support that ALK3 gene played an important role during heart development. The platelet-activating factors acetylhydrolase and Pax-8 genes could be important ALK3 downstream genes in the BMP signaling pathway during interventricular septum development. PTK and beta subtype protein 14-3-3 might be regulatory factors in this pathway.

Under simulated ultraviolet-B radiation (UV-B, 280-320 nm), effect of calcium on biological characteristics of wheat seedlings was studied with pot culture experiment. The results showed that the growth indexes of wheat seedlings on ground and underground in calcium concentration 2 mmol/L treated group were obviously reduced than that of wheat seedlings in control treated group (the descending range of 8.1%-36.1%) under UV-B stress (0.35 W/m2, 6 h per day, for 7 day). And the decreasing degree of growth indexes (3.2%-28.6%) for wheat seedlings in calcium concentration 6 mmol/L treated group were lower than in calcium concentration 2 mmol/L treated group. The experiment proved that calcium can mitigate the damage of ultraviolet-B radiation on wheat seedlings, and this effect was related to that calcium affected physiological and biochemical characteristics of wheat seedlings.

OBJECTIVE:To explore one method getting highly purified plasmid DNA and costing low.METHODS:According to principle of extraction and purification plasmid DNA of glass fiber chromatographic column, its operation sequence was improved so that disposable glass fiber chromatographic column could be used repeatedly. The concentration and purity of plasmid DNA obtained by 12 times repeated using glass fiber chromatographic column were determined by spectrophotometry. Moreover, Those plasmid DNA obtained were tested by restriction enzyme and agarose gel electrophoresis, Ligation in vitro and transformation test.RESULTS:12 times repeated use of glass fiber chromatographic column made no significant difference to the concentration and purity of obtained plasmid DNA (P < 0.05), and plasmid DNA was completely digested by restriction enzyme. The digested fragments were well ligated together by the action of T4 DNA Ligase in ATP ligating buffer in vitro and ligated DNA was well transformed into E. coli.CONCLUSION:Repeated use glass fiber chromatographic column can obtain a lot of highly purified plasmid DNA and cost is low. Digestion, ligation and transformation of those plasmid DNA are not affected. The results may be significant in molecular cloning and study of gene vaccination.