Background T helper cell 17 (Th17),a newly discovered subset of CD4+ T cells,have been found to play an important role in dry eye disease in animal model.Further investigation should be done on the immunopathogenesis of Th17 cells in dry eye patients.Objective This study was to analyze the expression status of interleukin-17 (IL-17) and IL-22 in tear and supernatant of peripheral blood mononuclear cells (PBMCs) of dry eye patients and their correlation with clinical symptom and sign.Methods Twenty Sj(o)gren syndrome (SS)patients,twenty non-Sj(o)gren syndrome (NSS) patients were included in Wuxi Second Hospital from 2010 to 2011,and twenty healthy volunteers were recruited at the same period.All of subjects understood the purpose and procedure of research and written informed consent was obtained form each subject initial of this study.Dry eye symptom questionnairs were self-answered and multiple dry eye disease-related clinical tests,including the breakup time of tear film (BUT),Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescein staining were performed.The periphery blood of 3 ml and tear were collected in all the subjects,and IL-17 and IL-22 levels in supernatant of PBMCs and tear were detected by enzyme-linked immunosorbent assay (ELISA).The correlations between levels of IL-17 and IL-22 with BUT,S Ⅰ t,corneal fluorescein staining and dry eye scores were analyzed.Results The dry eye scores reduced,BUT prolonged,S Ⅰ t increased and corneal fluorescein dye decreased from SS group,NSS group to normal control group,with significant differences among the three groups (dry eye scores:H =40.81,P＜0.01 ; BUT:H =40.15,P＜0.01 ;S Ⅰ t..H=50.07,P＜0.01 ;corneal dye scores:H=40.52,P＜0.01).The concentration of IL-17 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (964.92±124.83)ng/L,(718.85± 115.89)ng/L and (341.95±85.08) ng/L,showing a statistically significant difference among them (F=162.95,P＜0.01).The levels of IL-17 in the tear were (440.69±126.09) ng/L,(364.33±126.85) ng/L and (61.16±11.60) ng/L in the SS group,NSS group and normal control group respectively,exhibiting an elevated level in the SS group and NSS group compared with the control group (F=75.27,P＜0.01).In addition,the levels of IL-22 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (98.77± 11.27) ng/L,(79.65 ± 11.01) ng/L and (32.78±9.34) ng/L,and those in the tear were (22.22 ± 8.96) ng/L,(14.92 ±4.35) ng/L and (10.47 ± 2.67) ng/L,with significant differences among the three groups (F =206.27,P＜0.01 ;F =19.87,P＜0.01).The significant correlations were found between the IL-17 and IL-22 concentration in the supernatant of PBMCs and tear with corneal fluorescein staining scores and S Ⅰ t.Conclusions The contents of IL-17 and IL-22 in PBMCs and tear upregulate in the SS and NSS patients,indicating that Th17 plays a key role in the immunity mechanism of dry eye.

Background Mechanism of dry eye disease has not been established clearly.Some increasing evidences showed that inflammation plays an important role in pathogenesis and development of dry eye disease.However,the association of interleukin-6 (IL-6) with dry eye has not been clarified.Objective This study was to assess the correlation of IL-6 levels in tear and serum of patient with dry eye disease and clinical symptom.Methods Twenty patients with Sj（o）gren syndrome (SS) and 20 cases with non-Sj（o）gren syndrome (NSS) were enrolled from Second Affiliated Hospital of Nanjing Medical University,and 20 healthy volunteers were recruited as the normal controls.Symptom questionnaires were self-answered and multiple dry eye-related clinical tests were performed,including tear film breakup time (BUT),Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescence staining.IL-6 concentrations in serum and tear were detected by enzyme-linked immunosorbent assay (ELASA).The correlation between IL-6 level and tear film and ocular surface parameters was analyzed by Spearman rank correlation.Written informed consent was obtained from each subject initial of this trial.Results The scores of symptom questionnaires were 18.50 (11.75,23.00),23.00 (19.00,32.00) and 2.00 (0.25,4.00) in the SS group,NSS group and the normal control group,and significantly increasing values were seen in the SS group and NSS group compared with the normal control group (P=0.00,0.00).BUT values in the SS group and NSS group were 2.50 (2.00,3.88) seconds and 3.50 (2.13,4.00) seconds and were lower than 15.00 (13.13,17.00) seconds in the normal control group (P=0.00,0.00).S Ⅰ t values in the SS group,NSS group and the normal control group were 2.00 (1.50,2.88),4.00 (3.00,5.75) and 19.00 (18.00,25.00)mm,showing significant differences between the SS group and the normal control group (P =0.00),the SS group and the NSS group (P =0.00) or the NSS group and the normal control group (P=0.00).The marked elevation of keratoepithelioplasty score was seen between the SS group and the normal control group (5.75 (4.00,7.75) vs.0.00 (0.00,0.75) (P =0.00) or the NSS group and the normal control group (4.50(3.63,6.00) vs.0.00(0.00,0.75)) (P=0.00).Concentrations of IL-6 in serum and tear in the SS group,NSS group were (131.47±21.04) μg/L and (77.81 ± 15.68) μg/L and were significantly higher than (31.62±8.57) μg/L of the normal control group,and those in the tear were (33.44±8.01) μg/L,(18.78 ±5.73) μg/L and (8.77 ± 3.43)μg/L,respectively in the three groups,with a significant reduce in the normal control group (all P=0.00).tL-6 levels in serum and tear were positively correlated with the score of symptom questionnaires or keratoepithelioplasty and negatively correlated with BUT and S Ⅰ t (all P =0.00).Conclusions IL-6 levels in the tear and serum of the dry eye patient are associated with the severity of the disease.IL-6 levels in serum and tear may be objective,sensitive and reliable indicators of dry eye.

Objective To investigate the mechanism of the inhibitory effect of ?-ray irradiation on rat vascular smooth muscle cells(VSMC). Methods The ef fect of ?-ray irradiation on proliferation of VSMC was observed by 3?H-TdR incor poration. After ?-ray irradiation, the VSMC cell cycle change was detected b y flo w cytometry. The expression of p53, cyclin D and PCNA was investigated by Wester n Blot. Results The inhibitory effect of ?-ray irradiation on VSMC proliferati on was dose-dependent. After ?-ray irradiation, VSMC was arrested in G 1 st age, w ith the expression of p53 increased but the expression of cyclin D and PCNA decr eased. Conclusions ?-ray irradiation can inhibit the proliferation of VSMC. T he main mechanism is probably due to the induction of cell cycle arrest and inhi bition of the VSMC mitosis ,during which process, p53,cyclin D and PCNA all pla y an important role .

OBJECTIVETo investigate the effects of pregnancy malnutrition on the occurrence of insulin resistance (IR) in rat offspring during adult stage and to find out the relationship between TNF-alpha and IR; and to find out a reasonable early nutritional intervention measure for the prevention of IR, through giving different diets to offspring.

METHODSAn IUGR model was built by maternal nutrition restriction. 80 newborn IUGR female pups were randomly divided into 4 groups, the mother rats were given the following diet respectively for 3 weeks after delivery, pups were fed by mother milk: (1) The IUGR (intrauterine growth retardation) rat model was used and the animals were divided into: IUGR control group (group S/N) fed with normal diet, (2) IUGR high-caloric diet group (group A), (3) IUGR high-protein and high-caloric diet group (group B) and (4) IUGR high-protein isocaloric diet group (group C). Each group had 20 pups and another 20 normal female pups were fed with normal diet as the normal control group (group C/N). All pups were weaned at the 4th week of age and fed with normal diet till the end of the experiment. At the 12th week (adulthood) and 48th week (senility) of life, body weight and length, the fasting blood glucose, insulin concentration, TNF-alpha of adipose tissue and body weight were measured. Body mass index (BMI), ISI (insulin sensitive index), IRI (insulin resistant index) and HBCI (beta cell insulin excretion index) and their correlation to TNF-alpha were calculated.

RESULTSAt 12th week and 48th week of life, the insulin sensitivity of IUGR model group was significantly lower than group C/N, although there was no significant difference of body weight between these two groups. TNF-alpha was negatively correlated with ISI, positively correlated with IRI and no relation to HBCI. Group A and B was fatter and developed more severe IR. There were no significant differences in ISI, IRI, HBCI and TNF-alpha between group C and group C/N.

CONCLUSIONSIUGR offspring of pregnancy malnutrition mother rats showed IR at the age of 12th week. TNF-alpha was closely related to the occurrence of IR in IUGR pups. IUGR pups fed with high caloric diet or high protein and caloric diet at the early postnatal period amplified the metabolic abnormality. The high protein isocaloric diet is effective early nutritional intervention measure for the prevention of occurrence of IR at adulthood.

Animals ; Animals, Newborn ; growth & development ; Body Weight ; drug effects ; Dietary Proteins ; pharmacology ; Female ; Fetal Growth Retardation ; blood ; etiology ; Insulin Resistance ; physiology ; Malnutrition ; physiopathology ; Pregnancy ; Pregnancy Complications ; Prenatal Exposure Delayed Effects ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate effects of paraquat on the mRNA expression of key elements of Notch signaling (Notch1, Jagged1 and DTX1) during differentiation process of human neural stem cells (hNSCs).

METHODShNSCs exposed to PQ at the concentrations 0.10, 1.00, 10.00 M. Cell proliferation ability was assessed using MTT assay and mRNA expressions of Notch1, Jagged1 and DTX1 were detected by Real-time RT-PCR at 2, 4, 8, 12 d of differentiation.

RESULTSCompared with control group, NOTCH1, JAG1 mRNA expression levels exposed to PQ at the concentration of 0.10 M significantly reduced at 2, 4, 8 d and significantly went up at 12d (P < 0.01). Compared with control group, NOTCH1, JAG1 and DTX1 mRNA expression levels exposed to PQ at the concentration of 10.00 M significantly reduced at 2, 8, 12 d (P < 0.01). PQ could down-regulate Notch1, Jagged1 and DTX1 mRNA expressions at the early stage of differentiation, then up-regulate Notch1 mRNA expression, and down-regulate Notch1, Jagged1 and DTX1 mRNA expressions at the end of differentiation.

CONCLUSIONNotch signaling pathway may be involved in differentiation of neural stem cell exposed to PQ.

Calcium-Binding Proteins ; metabolism ; Cell Differentiation ; drug effects ; Cells, Cultured ; Embryonic Stem Cells ; cytology ; drug effects ; metabolism ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Jagged-1 Protein ; Membrane Proteins ; metabolism ; Neural Stem Cells ; cytology ; drug effects ; metabolism ; Paraquat ; pharmacology ; Receptor, Notch1 ; metabolism ; Serrate-Jagged Proteins ; Signal Transduction ; drug effects ; Ubiquitin-Protein Ligases ; metabolism

Adolescent ; Adult ; Ambulatory Surgical Procedures ; instrumentation ; methods ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; Radio Waves ; Tonsillectomy ; instrumentation ; methods ; Young Adult

Clinical epidemiologic data and animal experimental studies regard estrogen as being protective against the development of cardiovascular diseases. The mechanisms by which estrogen affects the development of vascular diseases are not clear. Recent studies demonstrated that the cardiovascular protective effects of estrogen are closely related to nitric oxide (NO) pathway. Our previous study proved that estrogen inhibited the proliferation and oncogene expression of vascular smooth muscle cells (VSMCs) induced by endothlin 1 (ET-1) and serum,this effect was mediated by NO release. In the present study, we investigated the role of inducible nitric oxide synthase (iNOS) in the VSMCs cycle arrest induced by 17 beta-estradiol (E(2)). The effects of E(2) on iNOS activity and protein expression in cultured rat VSMCs and the influence of NOS inhibitor N(G)-nitro-L-arginine methylester (L-NAME) on the inhibitory effect of E(2) on cell cycle were investigated. NOS assay kit was used to measure the activity of iNOS and protein expression of iNOS was determined by Western-blot. Cell cycle analysis was accessed by flow cytometry. The results obtained showed that E(2) increased iNOS activity of VSMCs but not in a dose-dependent manner. E(2) 10 nmol/L increased the iNOS activity of VSMCs distinctly at two time points: 30 min and 12 h. These effects were significantly inhibited by estrogen receptor (ER) antagonist Tamoxifen (0.1 micromol/L) and NOS inhibitor L-NAME (1 micromol/L). E(2) increased iNOS protein expression of VSMCs in a dose-dependent manner. The effect of E(2) on iNOS protein expression of VSMCs started at 3 h, distinctly increased at 12 h and then decreased. Tamoxifen significantly inhibited the E(2)-induced iNOS protein expression of VSMCs. ET-1 increased cell percentage of S phase and G(2)+S/G(1). This effect was inhibited by E(2). L-NAME significantly attenuated the inhibitory effect of E(2) on cell cycle of VSMCs. The results suggest that E(2) induced G(1) arrest of VSMCs, which was associated with an increase in iNOS activity and protein expression of VSMCs. These effects were at least mediated by estrogen receptor partly.
Animals ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; Cells, Cultured ; Endothelin-1 ; metabolism ; Estradiol ; pharmacology ; Estrogen Antagonists ; pharmacology ; Female ; Muscle, Smooth, Vascular ; cytology ; Nitric Oxide Synthase ; metabolism ; physiology ; Nitric Oxide Synthase Type II ; Rats ; Tamoxifen ; pharmacology

OBJECTIVETo observe the hair follicle regeneration after subcutaneous implantation of hair follicle cell mixture in nude mice.

METHODSThe hair papilla cells, dermal sheath cells, outer root sheath and fibroblasts of human scalp were mixed with the hair follicle epithelial cells and implanted subcutaneously in nude mice to observe the regeneration of the hair follicle.

RESULTS AND CONCLUSIONFormation of intact hair follicle-like structures was observed in the skin sections of the recipient nude mice, suggesting the feasibility of this approach for hair follicle regeneration in vivo.

Animals ; Cell Transplantation ; Hair Follicle ; cytology ; metabolism ; physiology ; transplantation ; Humans ; Injections, Subcutaneous ; Mice ; Mice, Nude ; Regeneration ; Skin Pigmentation ; Time Factors

Rat vascular smooth muscle cells (VSMC) were used to study the effect of 17beta -estradiol (E(2)) on cellular proliferation (cell counting), DNA synthesis ((3)H thymidine incorporation), MTT, c -fos mRNA expression and nitric oxide (NO) release. The results obtained showed that E(2) (10(-12) 10(-8) mol/L) induced NO release from VSMC in a concentration-dependent manner; 10(-8) mol/L E (2)significantly inhibited VSMC cellular proliferation and DNA synthesis induced by 10% FCS and 10(-7) mol/L ET-1, which was obviously reversed by 10(-7)mol/L tamoxifen and 10(-6) mol/L L-NAME; after a pretreatment for 24 hours, 10(-8)mol/L E(2) significantly inhibited VSMC c -fos mRNA expression induced by 10(-7)mol/L ET-1, which was also obviously reversed by 10(-6) mol/L L-NAME. These results suggest that the inhibitory effects of E(2) on VSMC cellular proliferation and c -fos mRNA expression are closely related with NO release in VSMC, which is, at least, partly medicated by ER.
Animals ; Cell Division ; drug effects ; Cells, Cultured ; Estradiol ; pharmacology ; Female ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Nitric Oxide ; metabolism ; physiology ; Proto-Oncogene Proteins c-fos ; biosynthesis ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Sprague-Dawley

The aim of this study was to determine the dynamic expression profile of hepatitis B surface antigen (HBsAg) according to hepatic parenchyma cells' volume at different stages of liver fibrosis during the immune clearance phase. Eighty-nine patients with HBeAg-positive chronic hepatitis B (CHB) in the immune clearance stage were recruited for study. Each patient's serum HBsAg levels were detected by electrochemiluminescence. The serum HBsAg levels were apportioned according to hepatic parenchyma cells' volume at liver fibrosis stages 1, 2, 3, and 4 and compared by ANOVA. The unapportioned serum HBsAg levels (IU/mL) at liver fibrosis stages 1 (227.2+/-237.7), 2 (211.0+/-131.4), 3(300.1+/-144.6), and 4 (278.7+/-148.8) were not significantly different (all comparisons, P range: 0.061 to 0.759). However, when the serum HBsAg levels were apportioned by the same hepatic parenchyma cells' volume at liver fibrosis stages 1 (343.9+/-359.8), 2 (336.4+/-209.5), 3 (508.7+/-245.1), and 4 (525.2+/-274.8), the levels were significantly different (all comparisons, F = 3.045 and P = 0.033; stage 1 vs. 3, P = 0.041; stage 1 vs. 4, P = 0.046; stage 2 vs. 3, P = 0.028; stage 2 vs. 4, P = 0.034). During the immune clearance phase of chronic hepatitis B, increased HBsAg expression is associated with increased hepatic parenchyma cells' volume and progressive liver fibrosis stage.
Adolescent ; Adult ; Cell Size ; Child ; Female ; Hepatitis B Surface Antigens ; blood ; Hepatitis B, Chronic ; metabolism ; pathology ; Humans ; Liver ; cytology ; metabolism ; Liver Cirrhosis ; metabolism ; pathology ; Male ; Middle Aged ; Young Adult