ObjectiveTo understand the pathological and physiological roles of Presenilin 1 (PS1) in Alzheimer's disease (AD) recurrence, and the interaction between PSI and carhoxyl terminus of Hsc70 interacting protein (CHIP). MethodsThe yeast two-hybrid system was applied to identify a novel PS1 interacting protein as CHIP. After pGBKT7-PS1-C203 bait plasmid and full fragement CHIP of pACT2-CHIP expression vector were constructed, the interaction between PSI and CHIP was tested by β-galactosidase assay, pGBKT7-PS1-C203 was co-transfected with pACT2-CHIP into 293T cells and the interaction between PS1 and CHIP was tested by co-immunoprecipitation and Western blot. ResultsSpecificity of the interaction between PS1 and CHIP was identified by β-galactosidase assay and co- immunoprecipitation. ConclusionsCHIP is able to modulate chaperone functions and the pathway of protein ubiquitination/degradation. CHIP may regulate a proper assembly of the γ-secretase complex through its interaction with PSI, which is helpful to elucidate the mechanism of AD pathology.

OBJECTIVEThis study investigated the life style and health status of preterm and term infants' mothers in order to explore the risk factors for preterm delivery.

METHODSA total of 600 matched cases, including 120 pairs of preterm infants and their mothers and 180 pairs of term infants and their mothers were recruited. All of the mothers participated in a questionnaire survey on life style and health status during or before pregnancy. General data of the infants were also collected. Logistics regression analysis was used for evaluating the risk factors for preterm delivery.

RESULTSThe height, body weights and body mass index before pregnancy in preterm infants' mothers were significantly lower than those in term infants' mothers (p<0.05). The birth height and weights and Apgar scores in preterm infants were significantly lower than those in term infants (p<0.05). The whole nutritional Knowledge-Attitude-Practice (KAP) scores in preterm infants' mothers were significantly lower than those in term infants' mothers (P<0.05). Multiple factor regression analysis showed that recurrent abortions before gestation (OR=2.332, P<0.05) and premature rupture of membrane before delivery (OR=7.979, P<0.01) were risk factors for preterm delivery, while maternal nutritional KAP scores (OR=0.949, P<0.01) and body weights before pregnancy (OR=0.954, P<0.05) were protective factors.

CONCLUSIONSThe women with low body weight, low nutritional KAP scores or recurrent abortions history before pregnancy or presenting premature rupture of membrane before delivery are at high risk for preterm delivery.

Case-Control Studies ; Female ; Fetal Membranes, Premature Rupture ; etiology ; Health Knowledge, Attitudes, Practice ; Health Status ; Humans ; Infant, Newborn ; Infant, Premature ; Life Style ; Logistic Models ; Pregnancy ; Premature Birth ; etiology

OBJECTIVETo study whether total saponins of Panax notoginseng (PNS) can prevent renal interstitial fibrosis occurrence through blocking the IL-1alpha induced tubular epithelial cell and decrease the secretion of extracellular matrix (ECM).

METHODSNormal rats' tubular epithelial cells NRK52E were cultured in vitro, their morphological changes were observed by inverted phase contrast microscope and scanning electron microscope. Flow cytometry technique and immuno-histochemical method was used to detect the expression of alpha-smooth muscle actin (alpha-SMA), and ELISA was used to quantitatively detect the fibronectin (FN) in the supernatant

RESULTSIL-1alpha could induce the transdifferentiation of tubular epithelial myofibroblast, showing hypertrophy of cells elongated and fusiform-shaped, with significantly enhanced expression of alpha-SMA and increased secretion of FN (P<0.05). After adding PNS of different concentrations, the morphology of cells restored close to normal tubular epithelial cells, with the increase of alpha-SMA expression and FN secretion significantly inhibited (P<0.05) in dose-dependent manner (P<0.05). But addition of different dosages of PNS alone showed no effect on tubular cells.

CONCLUSIONIL-1alpha could induce the transdifferentiation of tubular epithelial myofibroblast, promote the deposition of ECM component FN. PNS could inhibit IL-1alpha induced the transdifferentiation of NRK52E and secretion of ECM, therefore, PNS could be taken as a new drug for prevention and treatment of renal interstitial fibrosis and terminal stage of renal diseases.

Actins ; analysis ; Animals ; Cell Differentiation ; drug effects ; Cell Line ; Epithelial Cells ; cytology ; Fibronectins ; analysis ; Fibrosis ; prevention & control ; Interleukin-1 ; pharmacology ; Kidney ; pathology ; Kidney Tubules ; cytology ; Panax ; chemistry ; Rats ; Saponins ; isolation & purification ; pharmacology

Objective To investigate the role of interleukin-17 (IL-17) in spinal dorsal horns in neuropathic pain (NP) in rats and its effect on activation of astrocytes.Methods In vivo experiment Sixty-four male SPF Sprague-Dawley rats,aged 6-8 weeks,weighing 180-200 g,were randomly divided into 3 groups using a random number table:control group (group C,n =16),sham operation group (group S,n =24) and group NP (n =24).The animals were anesthetized with intraperitoneal pentobarbital sodium,the L5,6 spinal nerves of the left side of the rat were gently separated and exposed,tightly ligated with 5-0 silk suture and transected.In group S,the L5,6 spinal nerves of the left side of the rat were only exposed.In group C,no operation was performed.Mechanical pain threshold was measured at day 1 before operation and days 1,3,5,7,10 and 14 after operation.The expression of IL-17,IL-6,IL-1β and tumor necrosis factor-alpha (TNF-α) mRNA in the spinal dorsal horn was determined using quantitative real-time PCR at day 7 and day 14 after operation.At day 7 after operation,the activation of astrocytes in the spinal dorsal horn was detected.In vitro experiment Primarily cultured astrocytes of neonatal rats were randomly divided into 4 groups using a random number table:control group (group C,n=22),10 ng/ml IL-17 group (I10 group,n=18),50 ng/ml IL-17 group (I50 group,n-18) and 100 ng/ml IL-17 group (I100 group,n=22).In I10,I50 and I100 groups,the astrocytes were incubated with the culture medium containing 10,50 and 100 ng/ml IL-17,respectively.The proliferation of astrocytes was detected by MTT at 24,48 and 72 h of incutation or culture.The expression of IL-6,IL-1β and TNF-α mRNA was determined using quantitative real-time PCR.Results In vivo experiment Compared with group C,the mechanical pain threshold was significantly decreased at 3-14 days after operation,the expression of IL-17,IL-6 and IL-1β mRNA in the spinal dorsal horn was up-regualted at 7 days after operation,and the activation of astrocytes was increased in group NP,and no significant change was detected in the mechanical pain threshold at each time point after operation in group S.In vitro experiment Compared with group C,the proliferation of astrocytes was significantly increased at 48 h of incubation in I10 and I50 groups,the proliferation of astrocytes was significantly increased at 48 and 72 h of incubation,and the expression of IL-6 and IL-1β mRNA was up-regulated in I100 group,and no significant change was found in the proliferation of astrocytes in group S.Conclusion Up-regulated expression of IL-17 in spinal dorsal horns may be involved in the maintenance of NP,and the mechanism is related to promoted activation of astrocytes and induced inflammatory responses in rats.

Objective To study the alteration of Th17 cells and related molecules in patients with Hashimoto's thyroiditis (HT).Methods Th17 cells were determined by flow cytometry.Real-time PCR method was applied to detect the expression of the orphan nuclear receptor RORγt and interleukin(IL) -17.Serum IL-6 and IL-23 were detected by ELISA method.Results ( 1 ) Compared with healthy controls,the frequency of Th17 cells [ (0.75± 0.79) ％ vs ( 0.28 ± 0.23 )％ ] and expression of RORγt ( 0.30 ± 0.38 vs 0.04 ± 0.02,both P ＜ 0.05 ) were significantly increased in HT patients.( 2 ) The levels of IL-6 and IL23 in HT patients were higher than those in healthy controls ( 3.66 ± 4.70 vs 0.47 ± 1.11,154.7 ± 75.81 vs 80.65 ± 61.41,both P＜0.05 ).( 3 ) A positive correlation between Th17 cells and serum TgAb was revealed in HT patients ( r =0.848 4,P =0.007 7 ).(4) The results of PCR showed that IL-17 and RORγt expressed in thyroid tissues of patients HT.Conclusion An increased frequency of Thl7 cells was found in HT patients,implying that this cell subset may play an important role in the pathogenesis of autoimmune thyroid disease.

Objective:To explore the mechanism of B10 cell involved in cardiomyocyte hypertrophy following myocarditis, and to develop potential therapeutic strategies.Methods:BALB/c mice infected with Coxsackie virus B3 induced viral myocarditis model. The expression of angiotensin (ANG)Ⅱ and its receptor in myocarditis mice was detected. The changes of B10 cells in the hearts of control mice and myocarditis mice were analyzed by flow cytometry. After losartan was administered to myocarditis mice, the degree of myocardial inflammation was detected by HE staining, the expression of inflammatory factors was detected by ELISA, the myocardial hypertrophy was detected by wheat germ agglutinin (WGA) staining, and the changes of B10 cells in the heart were analyzed by flow cytometry. The levels of cardiac troponin T (C-TNT) and high mobility group box 1 (HMGB1) protein in neonatal mouse cardiomyocytes treated with ANGⅡ and ANGⅡ+ IL-10 were detected. Cardiomyocytes were treated with ANGⅡ, ANGⅡ+ B10 cells, ANGⅡ+ B10 cells + IL-10 receptor antibody and ANGⅡ+ B cells to detect C-TNT protein levels, and Annexin-V/PI was used to detect the apoptosis of cardiomyocytes. Cardiomyocytes were treated with oxidized HMGB1, reduced HMGB1 and disulfide HMGB1, and C-TNT expression was detected.Results:Coxsackievirus B3 infection caused cardiac hypertrophy, high expression of ANGⅡ and its receptor, and transient increase of B10 cells in mice. Losartan treatment blocked the angiotensin receptor, reduced expansion of B10 cells. B10 cells alleviated apoptosis of cardiomyocytes and inhibited the production of HMGB1 induced by ANGⅡ patch by producing IL-10, thus alleviating viral myocarditis and cardiac hypertrophy.Conclusions:B10 cells may play an important role in myocardial protection in myocarditis.

OBJECTIVETo explore the relationship between NB4 cell apoptosis induced by tanshinone IIA (TanIIA) and the cell mitochondrial transmembrane potential (DeltaPsim).

METHODSNB4 cells were treated with TanIIA, As(2)O(3), TanIIA plus 1.0 micro g/ml CsA and As(2)O(3) plus 1.0 micro g/ml CsA, respectively. Morphological changes were observed under light microscope and transmission electron microscope. The percentages of sub-G(1) cells and DeltaPsim of cells doublely stained with PI and Rh123 were assayed by flow cytometry.

RESULTSThe percentages of sub-G(1) cells after treatment with 1.0 micro g/ml and 2.0 micro g/ml TanIIA had no significant difference but was higher than that of 0.5 micro g/ml. After treatment with TanIIA, NB4 cells appeared the classical apoptotic morphology. The percentages of sub-G(1) cells were increased, while the DeltaPsim reduced (P < 0.01) and there was a linear correlation between them. The increment of sub-G(1) cell percentages and decrement of DeltaPsim induced by TanIIA were partly inhibited by CsA (P < 0.01).

CONCLUSIONSTanIIA can induce NB4 cells apoptosis through opening the mitochondrial permeability transition pore and reducing DeltaPSgr;m, and this effect could be inhibited by CsA.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; DNA ; metabolism ; Diterpenes, Abietane ; Humans ; Membrane Potentials ; drug effects ; Mitochondria ; drug effects ; Phenanthrenes ; pharmacology ; Tumor Cells, Cultured

OBJECTIVETo explore the expression of integrin alpha 6 in hepatic sinusoidal capillaration.

METHODSThe rat hepatic fibrosis model was established by injection of carbon tetrachloride subcutaneously. Then the expression of laminin and integrin alpha 6 subunit was observed by immunohistochemistry and dot immuno-blotting.

RESULTSWe observed sinusoidal capillaration formed by deposition of laminin along sinusoids in Disse interspace by immunohistochemistry staining. In normal rat the expression of integrin alpha 6 was restricted to portal vascular endothelial cells and bile duct epithelial cell membranes. No expression was observed in sinusoidal endothelial cell membranes. When capillaration integrin alpha 6 was detected in a continuous pattern along the sinusoids, the content of integrin alpha 6 was significantly higher in fibrotic liver tissues than in normal liver tissues as measured by dot immuno-blotting (P<0.05).

CONCLUSIONSDuring fibrogenesis, laminin continuously accumulate in liver tissues and form basement membrane resulting in sinusoidal capillaration, and then induce the expression of integrin alpha 6 on SEC membranes.

Animals ; Antigens, CD ; biosynthesis ; Carbon Tetrachloride ; Immunoblotting ; Immunohistochemistry ; Integrin alpha6 ; Laminin ; metabolism ; Liver ; metabolism ; pathology ; Liver Cirrhosis, Experimental ; chemically induced ; metabolism ; pathology ; Male ; Rats ; Rats, Wistar ; Time Factors

The coordination and collaboration of many links are required to achieve the successful development of a high-quality clinical randomized controlled trial, in which, the compliance of the researchers affects the ultimate results and quality of the study since it is responsible for the safety and benefits of subjects in the trials and study quality. Focusing on the researchers' compliance, it is believed that the improvements in the researchers' compliance are very significant in the implementation of study scheme. At present, commonly, the researchers are of the low compliance, manifest as failure to obey the study scheme (such as inclusive criteria, enrollment sequence), failure to obey the rules of operation and data collection, failure to store the original documents but to modify the data, failure to report timely adverse events, etc. In view of clinical research monitors, the reasons on the low compliance are analyzed and the solutions are proposed.

OBJECTIVETo set a quantitative criteria for determining the risks on cerebral vascular disease (CVD) so to identify that potential risk of an individual dying from CVD and to predict the individual risk of CVD.

METHODSData on case-control and cohort studies published during 1978 to 2003 was collected through retrieval of literatures, and data on surveillance of behavior exposure was provided by Chengdu Municipal Center for Disease Control and Prevention. Pooled odds ratio (OR) and relative risk (RR) of all risk factors for CVD were estimated using software for meta-analysis to enable the varied levels of risk factors be converted into risk fractions by statistical models.

RESULTSA risk score conversion table (quantitative criteria for assessment) of main risk factors for CVD was developed for men and women aged 35 - 69 at an interval of five years, including smoking, passive smoking, hypertension, high blood cholesterol levels, body mass index, lack of physical activity, alcohol drinking, dietary fat consumption, milk intake, oral contraceptive use, past history of diabetes and CVD, family history of CVD etc. Individuals with all these risk factors had a risk score beyond 1.00, but was equal to or below 1.00 when without. The risk score would increase along with the rise of one's risk level.

CONCLUSIONEstimation of risk of dying from CVD was based on risk score conversion table of risk factors for CVD, which could be used to predict individual potential risk of dying from CVD in the following 10 years. Our data provides evidence that education to be strengthened to persuade people to change their unhealthy lifestyles and behaviors.

Alcoholism ; complications ; Body Constitution ; Case-Control Studies ; Cerebrovascular Disorders ; epidemiology ; etiology ; China ; epidemiology ; Cohort Studies ; Contraceptives, Oral, Hormonal ; adverse effects ; Diabetes Complications ; Exercise ; Feeding Behavior ; Female ; Humans ; Hypertension ; complications ; Male ; Odds Ratio ; Risk Assessment ; Risk Factors ; Smoking ; adverse effects ; Urban Population ; statistics & numerical data