Up-regulated expression of Tim-3/Gal-9 at maternal-fetal interface in pregnant woman with recurrent spontaneous abortion

The relationship between T cell immunoglobulin domain and mucin domain protein 3（Tim-3）/Galectin（Gal）-9 pathway and recurrent spontaneous abortion（RSA） was studied. Thirty-one pregnant women with RSA and 27 normal early gravidas were investigated to detect the levels of Tim-3 and Gal-9 in villi and deciduas by Western blotting. Meanwhile, the concentration of interleukin（IL）-4 and IL-12 in peripheral blood plasma was determined by ELISA in 25 healthy fertile non-pregnant controls, the normal early gravidas and pregnant women with RSA mentioned above, respectively. It was found that the relative expression levels of Tim-3 and Gal-9 in villi and deciduas were significantly increased in pregnant women with RSA as compared with those in the normal early gravidas. The concentration of IL-4 in peripheral blood plasma of pregnant women with RSA was lower than that of the normal early gravidas（P〈0.05） and healthy fertile non-pregnant controls（P〈0.05）, but that of IL-2 in pregnant women with RSA was significantly higher than that of the normal early gravidas（P〈0.05） and healthy fertile non-pregnant controls（P〈0.05）. It was suggested that the overexpression of Tim-3/Gal-9 pathway may be related to the pathogenesis of RSA.     

Critical Amino Acid Residues for Nicotine 5＇ -Hydroxylation in Human CYP2A Enzymes

Objective： We have continued previous work in which we demonstrated that #117 and #372 amino acids contributed to the high activities of human CYP2A13 in catalyzing 4-methylnitrosamino-1-（3-pyridyl）-1-butanone（NNK） and aflatoxin BI（AFB1） carcinogenic activation. The present study was designed to identify other potential amino acid residues that contribute to the different catalytic characteristics of two CYP2A enzymes, CYP2A6 and CYP2A13, in nicotine metabolism and provide insights of the substrate and related amino acid residues interactions. Methods： A series of reciprocally substituted mutants of CYP2A6lle^300→ Phe, CYP2A6Gly^301aAla, CYP2A6Ser^369 → Gly, CYP2A13Phe^300→ Ile, CYP2A13Ala^301 → Gly and CYP2A13Gly^369 → Set were generated by site-directed mutagenesis/baculovirus-Sf9 insect cells expression. Comparative kinetic analysis of nicotine 5＇hydroxylatin by wild type and mutant CYP2A proteins was performed. Results：All amino acid residue substitutions at 300, 301 and 369 caused significant kinetic property changes in nicotine metabolism. While CYP2A6Ile^300→ Phe and CYP2A6Gly^301→Ala mutations had notable catalytic efficiency increases compared to that for the wild type CYP2A6, CYP2A13Phe^300→Ile and CYP2A13Ala^301→Gly replacement introduced remarkable catalytic efficiency decreases. In addition, all these catalytic efficiency alterations were caused by Vmax variations rather than Km changes. Substitution of #369 residue significantly affected both Km and Vmax values. CYP2A6Ser^369 → Gly increase the catalytic efficiency via a significant Km decrease versus Vmax enhancement, while the opposite effects were seen with CYP2A13Gly^369 → Ser. Conclusion：#300, #301 and #369 residues in human CYP2A6/13 play important roles in nicotine 5＇ -oxidation. Switching #300 or #301 residues did not affect the CYP2A protein affinities toward nicotine, although these amino acids are located in the active center. Set369 to Gly substitution indirectly affected nicotine binding by     

Comparison of Fresh Blood Cell Nature Immune Reaction of Children Patients with SLE or RA

To research the erythrocytes of condition of innate immunity in the children patients with RA and SLE, comparing with changes of activity of CD35 （CR1） on the erythrocyte between these two kinds of diseases, and further discuss the pathogenesis of these two kinds of diseases in children. The reaction of innate immunity of erythrocytes, lymphocytes and neutrophils which isolated from fresh blood of patients with RA and SLE to tumor cells was measured, and then the levels of expression of CD35 by using flow cytometry （FCM） were measured. The rosette rate of erythrocyte, lymphocytes and granulocytes adhering to cancer cell in the children patients with RA were higher than counterparts of normal controls （P〈0.001） respectively; the rosette rate of erythrocyte, lymphocytes and granulocytes adhering to tumor cells in the children patients with SLE were lower than counterparts of normal controls （P〈0.001） respectively. The rosette rate of these to tumor cells of children patients with RA, showed the highest figure change during different phases, including progression phase〉stationary phase 〉 extinction phase. And the rosette rates in these three groups were significantly higher than ones of that in normal controls. The expression of CD35 on the erythrocyte of children patients with RA was significantly higher than that of normal controls （P〈0.001）, while that of patients with SLE was significantly lower than that of normal controls （P〈0.001）. The CD35 on the red cells showed significantly positive correlation （γ=0.804, P〈0.01） with adhering erythrocytes and positively correlation with the rate of lymphocytes （P〈0.05）. The native immune function of children patients with SLE or RA was out of balance, and perhaps its changes have a relation with activity of diseases. It was suggested that the erythrocyte immunity function may be a new indicator to the condition of children patients with RA or SLE to judge the development and prognosis.     

Effects of IL-17 on expression of GRO-α and IL-8 in fibroblasts from nasal polyps

Recent studies indicated that interleukin（IL）-17, growth-related oncogene（GRO）-α and IL-8 play an important role in the pathogenesis of nasal polyps. However, the effects of the increased amount of IL-17 and the production of GRO-α and IL-8 in human nasal polyp fibroblasts are not completely understood. This study aimed to determine the effects of the increased IL-17 on the changes of GRO-α and IL-8 expression in human nasal polyp fibroblasts and further investigate the mechanism of neutrophil infiltration in nasal polyps. Nasal polyp fibroblasts were isolated from six cases of human nasal polyps, and the cells were stimulated with five different concentrations of IL-17. Real-time fluorescence quantitative polymerase chain reaction（RT-PCR） was used to detect the mRNA expression of GRO-α and IL-8. The mRNA of GRO-α and IL-8 was expressed in unstimulated controls and remarkably increased by stimulation with IL-17. Moreover, the levels of GRO-α and IL-8 produced by fibroblasts were increased gradually with the increases in IL-17 concentrations. The present study showed that nasal fibroblasts can produce GRO-α and IL-8, and their production is remarkably enhanced by IL-17 stimulation, thereby clarifying the mechanism of the IL-17 mediated neutrophil infiltration in nasal polyps. These findings might provide a rationale for using IL-17 inhibitors as a treatment for nasal inflammatory diseases such as nasal polyps.     

^131Ⅰ Radioactive Seeds for Tumor Bearing Mice of U87

A study was performed to make different activities ^131Ⅰ radioactive seeds for tumor bearing mice of U87. The ^131Ⅰ radioactive seeds of 89, 115, 185, 340MBq were sealed in soft plastic tube. Absorbed dose were measured by film and were calculated by S value which was made by Monte Carlo. When the tumor of U89 appeared in the nude mice, these seeds were implanted near the tumor. Then the size of＇tumor was measured and the curves of tumor growth were made. After all of these, the continuous low dose rate radiation of long time by ^131Ⅰwas discussed.     

Sphingosine Kinase-1/sphingosine 1-phosphate pathway in diabetic nephropathy

Objective Diabetic nephropathy （DN） is the major cause of end-stage renal disease worldwide and its prevalence continues to increase.Currently,therapies for DN provide only partial renoprotection; hence new targets for therapeutic intervention need to be identified.In this review,we summarized the new target,sphingosine kinase-1/sphingosine 1-phosphate （SphK1/S1P） pathway,explored its potential therapeutic role in the prevention and treatment of DN.Data sources Most relevant articles were mainly identified by searching PubMed in English.Study selection Mainly original articles and critical review articles by major pioneer investigators in this field were selected to be reviewed.Results SphK1/S1P pathway can be activated by hyperglycemia,advanced glycation end products,and many proinflammatory cytokines,which leads to fibronectin,transforming growth factor-31 up-regulation and AP-1 activation.And then it could promote glomerular mesangial cells proliferation and extracellular matrix accumulation,mediating the initiation and progression of diabetic renal fibrosis.Conclusions SphK1/S1P pathway is closely correlated with the pathogenesis of DN.The results suggest that SphK1/ S1P pathway as a new target for clinically improving DN in future is of great prospect.     

Electroacupuncture at Feishu （BL13） and Zusanli （ST36） down-regulates the expression of orexins and their receptors in rats with chronic obstructive pulmonary disease

Inflammation and lung function decline are the main pathophysiological features of chronic obstructive pulmonary disease （COPD）. Acupuncture can improve lung function in patients with COPD, but the underlying mechanisms are not well understood. Orexins （OXs）, which are found in peripheral plasma, are neuropeptides that regulate respiration and their levels are related to COPD. Therefore, we hypothesized that acupuncture might alter OXs, reduce lung inflammation and improve lung function in COPD. METHODS： COPD was induced in rats by exposure to cigarette smoke for 8 weeks and injecting with lipopolysaccharide twice. Electroacupuncture （EA） was performed at Feishu （BL13） and Zusanli （ST36） for 30 min/d for 2 weeks. Rat lung function and morphology were assessed after EA. The levels of tumor necrosis factor-α （TNF-a） and intedeukin-1β （IL-16） in bronchoalveolar lavage fluid （BALF） and orexin A and B levels in the lung tissue were detected by enzyme-linked immunosorbent assay. OX receptor mRNA levels and immunopositive cells were assessed with real-time polymerase chain reaction and immunohistochemical methods, respectively. The relationships among lung function, cell factors, and OX levels were analyzed by Pearson correlation analyses. RESULTS： Compared with the control group, lung function was significantly decreased in the rats with COPD （P〈0.05）. There were obvious increases in TNF-α and IL-1β levels in BALF （P〈0.05 and P〈0.01, respectively）, orexin A level in lung tissue （P〈0.01; but not orexin B） and mRNA expressions of OX 1 type receptor （OXR1） and OX 2 type receptor （OXR2） in lung tissue （P〈0.05 and P〈0.01, respectively）; the integrative optical densities （IODs） of both receptors were greater in the COPD group （P〈0.05）. For rats with COPD subjected to EA, lung function was improved （P〈0.05）. There were notable decreases in TNF-a and IL-1β levels （P〈0.05 and 〈0.01, respectively） in BALF. Orexin A, but not orexi     

MiR-503 regulates cisplatin resistance of human gastric cancer cell lines by targeting IGF1R and BCL2

Background Studies have shown that the drug resistance of gastric cancer cells can be modulated by abnormal expression of microRNAs （miRNAs）.We investigated the role of miR-503 in the development of cisplatin resistance in human gastric cancer cell lines.Methods MiR-503 expression was measured by quantitative real-time PCR.MTT （3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide） and clonogenic assays were used to examine changes in cell viability and the drug resistance phenotype of cancer cells associated with upregulation or downregulation of the miRNA.A dual-luciferase activity assay was used to verify target genes of miR-503.Immunohistochemistry,Western blotting analysis,and a flow cytometric apoptosis assay were used to elucidate the mechanism by which miR-503 modulates drug resistance in cancer cells.Results MiR-503 was significantly downregulated in gastric cancer tissues and several gastric cancer cell lines.Additionally,downregulation of miR-503 in the cisplatin （DDP）-resistant gastric cancer cell line SGC7901/DDP was concurrent with the upregulation of insulin-like growth factor-1 receptor （IGF1R） and B-cell lymphoma 2 （BCL2） expression compared with the parental SGC7901 cell line.An in vitro drug sensitivity assay showed that overexpression of miR-503 sensitized SGC7901/DDP cells to cisplatin.The luciferase activity of reporters driven by IGF1R and BCL2 3＇-untranslated regions in SGC7901/DDP cells suggested that IGF1R and BCL2 were both direct target genes of miR-503.Enforced miR-503 expression in SGC7901/DDP cells reduced expression of the target proteins,inhibited proliferation,and sensitized the cells to DDP-induced apoptosis.Conclusion Our findings suggest that hsa-miR-503 modulates cisplatin resistance of human gastric cancer cells at least in part by targeting IGF1R and BCL2.     

Effect of Mizolastine on Release of Inflammatory Mediators in Sensitized Mice

Mizolastine is widely prescribed for acute allergic cutaneous diseases because of its powerful antihistamine activity. Moreover, it has been recently put forward that mizolastine might also be effective on other inflammatory and refractory disorders such as atopic dermatitis and psoriasis. The aim of the present study is to investigate the effect of mizolastine on the release of two kinds of inflammatory mediators and cytokines （Leukotriene B4 and Interleukin-5）, which play pivotal roles not only in chronic allergic diseases but also in inflammatory diseases. The study was performed using epidermis from mice stimulated by intradermal injection of substance P and splenocytes from mice sensitized by intraperitoneal administration of ovalbumin （OVA）. Competitive enzyme-linked immunosorbent assay （ELISA） method was applied to examine the effects of mizolastine and the control drugs （chlorpheniramine, rolatadine and dexamethasone） on the release of Leukotriene B4 （LTB4） and Interleukin-5 （IL-5）. Mizolastine inhibited the release of LTB4 and IL-5 in epidermis as well as in splenocytes. The inhibitory effect was dose-dependent. Neither chlorpheniramine nor rolatadine showed inhibitory effects release from epidermis and splenocytes by mizolastine was not as strong as that by dexamethasone. Nevertheless, the results showed inhibitory effect of mizolastine over leukotriene and Th2 cytokines other than its anti-histamine effect. Thus, mizolastine seems to regulate on the release of inflammatory mediators and to have the potential to reorientate the T-cell response toward a balance between Th1 and Th2, which might have implications for the treatment of other inflammatory and chronic disorders.     

益智健脑颗粒联合针灸对阿尔茨海默病大鼠学习记忆的影响

目的观察益智健脑颗粒联合针灸对阿尔茨海默病（Alzheimers disease,AD）大鼠学习记忆的影响。方法将大鼠随机分为假手术组（A组）、模型组（B组）、针灸组（C组）、益智＋针灸组（D组）各10只,B、C、D 3组分别以海马CA1区注射β淀粉样蛋白25-35（Aβ25-35）造模,A组注射等量的双蒸水,各组分别治疗20 d后行Morris水迷宫试验,观察大鼠学习记忆能力变化。结果B组较A组的平均潜伏期明显延长,差异具有统计学意义（P〈0.05）;与B组比较,C组、D组的平均潜伏期明显缩短,过台次数增多,差异具有统计学意义（P〈0.05,P〈0.01）;与C组比较,D组的潜伏期缩短,过台次数增多,差异具有统计学意义（P〈0.05）。结论益智健脑颗粒联合针灸能够提高Aβ25-35介导的AD模型大鼠的学习记忆能力。     

Inhibition of interleukin-13 gene expression in T cells through GATA-3 pathway by arsenic trioxide

Aarsenic trioxide （AT） has a long history of use in both traditional Chinese medicine and in modem medicine in asthma therapy. Recently, Yin et al＇ found that AT even at small doses reduced the airway inflammation of sensitized guinea pigs. However the mechanism underlying this is still largely unknown. Interleukin 13 （IL-13）, as one of the important TH2 cytokines, plays an important role in asthma pathogenesis through promoting eosinophilic inflammation,     

The evaluation of Tracp5b as a marker for monitoring treatment results of bone metastasis in breast cancer patients

Objective ：To evaluate the sensitivity of serum tartrate-resistant acid phosphatase 5b（Tracp5b） activity in monitoring bisphosphonate treatment results of bone metastasis in breast cancer（BC） patients. Methods：The serum activities of Tracp5b, CEA, CA153 were measured in 58 BC patients, including 26 without bone metastasis, 32 with bone metastasis. The serum activities of TracpSb, CEA, CA153 were also measured in 19 patients with bone metastasis after 3 months of bisphosphonate treatment. Eighteen healthy women with age from 34 to 70 served as control. Results：Serum TracpSb was significantly elevated in patients with bone metastasis compared with that in all any other groups（P〈 0.05）. The sensitivity of TracpSb was 78.13% and the specificity was 86.36%. The sensitivity of CA153 was 37.50% and the specificity was 77.27%. The sensitivity of CEA was 21.88% and the specificity was 84.09%. The serum activity of TracpSb decreased significantly（P 〈 0.05） after 3 months of bisphosphonate treatment, while the levels of CA153 and CEA were unchanged. Conclusion：Serum Tracp5b activity is a useful diagnostic marker for bone metastasis in BC patients and can be used to evaluate the treatment results of bisphosphonate.     

Clinical study of electro-acupuncture treatment with different intensities for functional constipation patients

Functional constipation （FC） is a common functional bowel disorder disease that affects life quality of a large number of people. This study aimed to explore the impact of different intensities of electro-acupuncture （EA） treatment for FC patients. Totally, 111 patients with FC meeting the Rome III criteria were randomly assigned to different intensities of EA groups （low and high intensity of EA groups） and medicine-controlled （MC） group. In EA groups, patients were treated with EA at quchi （Llll） and shangjuxu （ST37） bilaterally for 4 weeks, 5 times/week in the first 2 weeks, and 3 times/week in the last 2 weeks. In MC group, 5 mg mosapride citrate was administered orally 3 times/day for 4 weeks. Spontaneous bowel movement frequency each day was recorded using a consti- pation diary. Self-rating anxiety scale （SAS） and self-rating depression scale （SDS） were used to assess the patients＇ psychological state. Cortisol （CORT）, substance P （SP）, and vasoactive intestinal polypep- tide （VIP） were evaluated at baseline and at the end of 4 weeks after treatment. As compared with the baseline, there was statistically significant increase in stool frequency every week （P〈0.01）, but there was no statistically significant difference among the three groups. As compared with the baseline, after 4 weeks of EA therapy, the scores of SDS and serum levels of CORT were decreased significantly in low intensity of EA group （P〈0.01）, and the serum levels of SP and VIP were increased significantly （P〈0.05）; the scores of SAS and SDS and serum levels of CORT were decreased significantly in high intensity of EA group （P〈0.05）, and the serum levels of SP and VIP were increased significantly （P〈0.05）; the serum levels of CORT and VIP were increased significantly in MC group （P〈0.05）. As compared with MC group, after 4 weeks of treatment, the serum levels of SP were signifcicantly in- creased in low intensity of EA group （P〈0.01）. Low and high intensi     

Sperm DNA Fragmentation may Influence IUI Outcome but could be Treated by ICSI： Evidence from Human Sperm Bank

Objective To determine the correlation between semen parameters, sperm DNA damage, progressive motility （PR）, morphology and intrauterine insemination （IUI）/ intracyto- plasmic sperm injection （ICSI） outcomes. Methods All the donors providing the samples in this study were recruited by Shang- hai Human Sperm Bank. For IUI, 122 donors were divided into group A （n=60） and group B （n=62）. Group A had a higher pregnancy rate while group B had a lower pregnancy rate （3.86 ± 1.50% vs 0.18 ± 0.52%）. For ICSI, 45 donors were divided into group C with a higher pregnancy rate （77. 78 ± 17.21%, n=23）, group D with a lower pregnancy rate （40. 73 ± 19.19%, n=22） and group E with an average preg- nancy rate in the sperm bank （48.96 ± 12.08%, n=23）. Semen analysis, morphology and DNA damage were assessed on samples retained in the sperm bank. Fresh semen samples were also collected and corresponding semen analyses data was included along with the pregnancy rates. Results No significant difference was found in the population characteristics between groups A and B, while there was a significant difference in sperm DNA fragmetation index （DFI） and morphology between the two groups （P〈0.05）, but not in other semen parameters （P〉0.05）. There was no significant difference in population characteristics between groups C, D and E while the DFI of group D was significant higher than groups C and E （P〈0.01）, but there was no significant difference in other semen parameters （P〉0.05）.Conclusion DFI might be a good predictor for IUI outcomes. Infertile couples with a high DFI should choose ICSI treatment instead of IUI. DFI should be a routine screening marker used to screen for sperm donors.     

Extraction of Evoked Potentials Based on Borel Spectral Measure for Less Trial Mixtures in Stable Distribution Environments

Stable process can better model the EEG noises found in the impact acceleration experiment. A new method for identifying the independent components of evoked potentials for less trial mixtures is proposed. The method is based on an estimate of the discrete Borel spectral measure for the characteristic function of an alpha stable random vector. Simulations demonstrate that the proposed method can identify independent components and the basis vectors of mixing matrix in the so-called less trial mixtures case of more sources.     

支气管哮喘患者白细胞介素12、免疫球蛋白E及内皮素1的水平测定及意义

支气管哮喘是临床常见的呼吸道疾病，是呼吸道慢性变态反应性炎症性疾病。气道慢性变态反应性由多种炎性细胞、炎性介质和细胞因子共同参与，检测哮喘患者自细胞介素（interleukin，IL）-12、免疫球蛋白E（immunoglobulin—E，Ig—E）及内皮素1（endothelin-1，ET-1）水平，以探讨三者在支气管哮喘不同病程中的作用。     

Anti-helicobacter pylori effect of total alkaloids of sophora alopecuroides in vivo

Background Helicobacterpylori （H.pylori） infection could lead to most gastroduodenal diseases and is even identified as a carcinogen of gastric cancer.Total alkaloids of sophora alopecuroides （TASA） is widely used in herbal remedies to treat various infectious diseases,including stomach-associated diseases.This study is aimed at evaluating the antimicrobial activity of TASA on H.pylori-infected BALB/c mice mouse gastritis.Methods Totally 120 BALB/c mice were orally inoculated with H.pylori Bacterial liquid to construct BALB/c mice H.pylori infection gastritis animal model,after the model was successfully created.We randomly assigned 100 infected mice into 10 treatment groups,the first group （normal saline）; the second group （bismuth pectin）; the third group （omeprazole）; the fourth group （TASA 2 mg/d）; the fifth group （TASA 4 mg/d）; the sixth group （TASA 5 mg/d）; the seventh group （TASA ＋ bismuth pectin）; the eighth group （TASA ＋ omeprazole）; the ninth group （bismuth pectin ＋ clarithromycin ＋ metronidazole）;the tenth group （omeprazole ＋ clarithromycin ＋ metronidazole）,5 other non-infected mice as negative control.Mice were orally inoculated twice a day and 7 days continuously.Then the mice were killed 4 weeks after treatment,we used realtime PCR to detect 16sDNA of H.pylori to test both the colonization and the clearance mice of bacteria of each treatment.We applied hematoxylin and eosin （HE） staining and immunostaining of mice gastric mucosa to observe the general inflammation and related factors interleukin 8 （IL-8）,cyclooxygenase 2 （COX-2）,and nuclear factor-kappa B （NF-KB） expression change after treatments.Results Firstly,we ensured that after 6-week intragastric administration,the bacteria colonization reached an exceed peak which is far higher than positive threshold （P ＜0.001）; secondly,after treatments,it is revealed that TASA combined with omeprazole or bismuth pectin showed promising antimicrobial activity against H.pylor