Aim To investigate the effect of GW0742 on the endothelial dysfunction induced by high glucose
(glucose at 55 mmol · L -1 )in isolated rat thoracic aorta and its related mechanisms.Methods The end
othelium-dependent relaxation of acetylcholine was per-formed in the absence or presence of GW0742 at differ-ent concentrations under high glucose condition.The structure of aorta was observed by HE staining.Moreo-ver,the content of NO was also measured by nitrate re-duction method.The mRNA and protein expression were detected by quantitative real-time PCR and West-ern blot,respectively.Results Compared with the control group,acetylcholine-induced vasodilatation was impaired by high glucose.Meanwhile,the structures of endothelial cells and smooth muscle cells were also in-terrupted.Furthermore, the expressions of PPARβmRNA and protein reduced while the NF-κB p65 ex-pression increased significantly which occurred in par-
allel with decreasing eNOS expression and NO concen-tration (P <0.01 ).GW0742 (0.01 ,0.1 ,1 μmol· L -1 )restored the relaxation of acetylcholine in a dose-dependent manner,and reversed the mRNA and pro-tein expression of PPARβ,NF-κB p65 and eNOS,as well as NO content (P <0.01 ).Conclusion GW0742 attenuates the injury of endothelial dysfunc-tion induced by high glucose,which may be,at least partly,mediated by the up-regulation of PPARβ,then the down-regulation of NF-κB,and the activation of eNOS-NO signal pathway.

Objective To investigate the effect of Shanzha Xiaozhi Capsules (SXC) on body weight, metabolic indexes, body fat accumulation and distribution of obesity patients with phlegm-dampness constitution. Methods Fifty obesity patients with phlegm-dampness constitution were randomized into control group and medicine group, 25 cases in each group. The control group was given lifestyle instructions for diet and exercises, and the medicine group was treated with SXC orally besides the instructions for lifestyle. The treatment for both groups covered 12 weeks. Before and after treatment, we observed the outcomes including scores of phlegm-dampness, body weight, waist circumference, hip circumference, blood pressure, fasting blood glucose, serum lipid profile, urine acid, body fat accumulation and distribution, and visceral fat accumulation. Results(1) The scores of phlegm-dampness were decreased significantly in both groups after treatment (P<0.05 or P<0.01), and the decrease was obvious in the medicine group(P<0.01).(2) Body weight and BMI were decreased significantly in both groups after treatment(P<0.01), and the decrease was obvious in the medicine group(P<0.05). Waist circumference was decreased significantly in the medicine group(P <0.05) but stayed unchanged in the control group(P>0.05). Hip circumference was decreased significantly in both groups(P <0.05 or P<0.01) , but the difference between the two groups was insignificant(P>0.05).(3) Systolic pressure and diastolic pressure were decreased significantly in both groups (P<0.01). But the difference of blood pressure decrease between the two groups was insignificant(P > 0.05).(4) Fasting blood glucose, 2-hour blood glucose after oral glucose tolerance test, serum lipid profile and urine acid remained unchanged in both groups after treatment (P>0.05). (5) The general body fat accumulation of both groups was decreased significantly after treatment (P <0.05 or P<0.01), and the decrease was obvious in the medicine group(P<0.01). Body fat accumulation in legs, trunk and gynoid region were decreased significantly in both groups after treatment(P<0.05 or P<0.01), but the difference of fat accumulation decrease between the two groups was insignificant(P>0.05). Waist fat accumulation had no significant decrease in the control group (P > 0 . 05), but was decreased in the medicine group (P<0.01). Visceral fat accumulation was decreased significantly in both groups after treatment(P<0.01), and the decrease was obvious in the medicine group (P<0.01). Conclusion In addition to its contribution to body weight loss, SXC also contribute to the reduction of visceral fat accumulation in obesity patients with phlegm-dampness constitution .

Objective To analyze the clinical infection distribution and drug resistance status of 442 strains of Escherichia coli to provide the basise for the treatment of Escherichia coli infection and the control of nosocomial infection.Methods The clinically submitted various kinds of specimens during 2013 were performed the bacterial culture and identification.The susceptibility of Escherichia coli to commonly used bacterial drugs were detected by adopting the MIC method.The data were analyzed by WHO-NET V5.5 and SPSS V13.0 softwares.Results 442 strains of Escherichia coli were isolated from the middle urine and secretion. The detection rate of ESBLs-producing Escherichia coli was 61.3%.442 strains of Escherichia coli had the high resistance to peni-cillins,cephalosporins and fluoroquinolones,better sensitivity toβ-lactam/β-lactamase inhibitor compounds and highest sensitivity to carbapenems.ESBLs-producing Escherichia coli had the higher resistance to commonly used antibacterial drugs than non-ESBLs-producing Escherichia coli .Conclusion The drug resistance of Escherichia coli is severe.ESBLs-producing Escherichia coli are u-sually resistant to many different types of antimicrobial drugs.Carbapenems are the first choice to treatment of severe infections of Escherichia coli .

Objective To investigate the effects of Shanzha Xiaozhi capsule on body weight, metabolic index, percentage and distribution of body fat in obese patients with dampness-heat constitution. Methods A total of 60 obesity patients with dampness-heat constitution were enrolled in this study and randomly categorized into the control group ( n=30) and treatment group (n=30). Control group was given the diet and exercise regime, and treatment group was given Shanzha Xiaozhi capsule based on the same diet and exercise regime for 12 weeks. Scores of dampness-heat, body weight, waist circumference, hip circumference, blood pressure, blood glucose, serum lipid profile, urine acid (UA), fasting plasma insulin, serum levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, adiponectin, lipopolysaccharide (LPS), percentage and distribution of body fat were measured before and after treatment. Results (1) Data of scores of dampness-heat and diastolic blood pressure were significantly decreased after treatment in drug group. The above indexes and hip circumference were significantly decreased after the treatment in treatment group compared with those of control group ( P<0.05). (2) The triglyceride (TG), UA, insulin, HOMA-IR, LPS and IL-1βwere significantly decreased after the treatment in treatment group than those of control group, while there were no significant differences in fast blood glucose (FBG), 2 h postprandial plasma glucose (2 hPG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), TNF-α, IL-6 and adiponectin between two groups. (3) The percentage of visceral fat was significantly decreased in treatment group than that of control group. There were no significant differences in fat changes of limbs and buttocks between two groups. Conclusion Shanzha Xiaozhi capsule can decrease body weight and visceral fat content, reduce insulin resistance, and improve chronic inflammation state in obese patients with dampness- heat constitution.

Objective To evaluate the knowledge,attitude and practice(KAP)related with AIDS of local floating population.Methods A total of 1 942 floating persons were sampled by multistage cluster sampling and interviewed with questionnaire in the agricultural markets,construction sites,and their habitats within 3 communities in Hongkou District.Results 82.9% of the interviewees were 20 ～49 years old.The average score was 34.77 ±3.52 (maximum score is 44)for knowledge,12.11 ±2.32(maximum score is 19)for attitude,5.50 ±0.95(maximum score is 7)for practice,respectively.The scores increased with educational level significantly.The scores of knowledge were 33.26 ±3.54,34.63 ±3.23,36.56 ±3.39 among the subjects with educational levels of primary school and below,junior high school,senior high school and above.The scores of attitude were 13.77 ±2.27,14.79 ±2.39,15.62 ±2.38,respectively.And the score of practice was 5.40 ±0.90,5.51 ±0.93,5.58 ±1.03,respectively.Conclusion In the present,the KAP relating AIDS of local floating population is poor in Hongkou District.A variety of intervention methods and operational patterns should be developed for the floating population with different educational levels and jobs.

OBJECTIVETo investigate the mechanism for the apoptosis of hippocampus neuron induced by hypothyroidism in perinatal rats.

METHODSHypothyroidism was induced by administration of propylthiouracil (PTU, 50 mg/d) solution to the dams from gestational day 15 by gavage. Pups from both hypothyroid and control groups were harvested at 1, 5, 10 and 15d, respectively. Blood samples were collected at the time of death for the determination of thyroid hormone. Serum free triiodothyronine (FT(3)) and free thyroxine (FT(4)) were measured by chemoluminescence. Hippocampus specimens were collected from the control and hypothyroid pups.Mitochondia was examined under transmission electron microscopy. Translocation of apoptogenic molecules (Bax, cytochrome C and AIF) and activation of caspase-3 were analyzed by Western Blotting.

RESULTSignificantly low circulating FT(3) and FT(4) levels confirmed the hypothyroid status of the experimental pups. Electron microscopy showed that altered morphology of mitochondria significantly increased under hypothyroid conditions. The expression of Bax in the cytosol of hypothyroid pups was higher than that of control pups at all stages of development (P<0.05),and significantly higher in mitochondria (P<0.001). The expression of cytochrome c in the cytosol of hypothyroid pups was significantly higher than that of control pups at all stages of development (1,10 and 15 d:P<0.05, 5d: P<0.001), and lower in mitochondria (P<0.05). The expression of AIF in the cytosol of hypothyroid pups was higher than that of control pups at all stages of development (P<0.001), and significantly lower in mitochondria (1, 5d: P<0.001, 10, 15 d: P<0.01). he expression of caspase-3 P20 in the cytosol of hypothyroid pups was significantly higher as compared with that of the age-matched controls (1, 15d: P<0.01, 5,1 0 d: P<0.001).

CONCLUSIONThe intrinsic death pathway in mitochondria may be one of the mechanisms with which hypothyroid induces apoptosis of hippocampus neuron in developing rats.

Animals ; Animals, Newborn ; Apoptosis ; physiology ; Female ; Hippocampus ; pathology ; Hypothyroidism ; chemically induced ; pathology ; Neurons ; pathology ; Pregnancy ; Pregnancy Complications ; pathology ; Propylthiouracil ; Rats ; Rats, Wistar

OBJECTIVETo investigate the influence of lead exposure in rats during the developmental stage on the expression of leptin in plasma, cerebrospinal fluid, and hippocampus, as well as investigating whether leptin is associated with the mechanism of cognitive impairment induced by lead exposure.

METHODSThe rat model of cognitive impairment after chronic lead exposure was established by adding lead acetate into drinking water. According to the concentration of lead acetate in drinking water, the rats were divided into control (0 ppm), low-lead (50 ppm), medium-lead (200 ppm), and high-lead groups (1 000 ppm), with 16 rats in each group. Atomic absorption spectrometry was used to measure the content of lead in the plasma, cerebrospinal fluid and hippocampus. ELISA was used to measure the level of leptin in the plasma and cerebrospinal fluid. Immunohistochemistry was used to observe the distribution of leptin protein in the hippocampus. Western blot was used for relative quantification of leptin proteins in the hippocampus.

RESULTSCompared with the control group, the lead exposure groups showed significant increases in the content of lead in blood, cerebrospinal fluid, and hippocampus (P<0.01), as well as significant reductions in the levels of leptin in plasma and cerebrospinal fluid (P<0.05). The results of immunohistochemical staining showed that leptin was mainly distributed in the cytoplasm of pyramidal neurons in the hippocampal CA region. The results of Western blot showed that compared with the control group, the three lead exposure groups showed a slight increase in the protein expression of leptin in the hippocampus (P>0.05).

CONCLUSIONSLead exposure can reduce the levels of leptin in plasma and cerebrospinal fluid in rats, which may be associated with the mechanism of cognitive impairment induced by lead exposure.

Animals ; Apoptosis ; drug effects ; Cognition ; drug effects ; Female ; Hippocampus ; chemistry ; drug effects ; pathology ; Lead ; blood ; toxicity ; Leptin ; analysis ; blood ; cerebrospinal fluid ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVEKawasaki disease (KD) is an acute febrile vasculitic syndrome of unknown etiology that preferentially affects coronary artery. It has been suggested that proinflammatory cytokines like tumor necrosis factor alpha (TNF-alpha) and interleukin-10 (IL-10) are key players during acute KD. Recently, the polymorphisms relative to major transcriptional start site of TNF-alpha and IL-10 gene were shown to influence the level of TNF-alpha and IL-10 production in vitro. This study was aimed to investigate the genetic association of TNF-alpha and IL-10 promoter polymorphisms in juvenile patients of Han nationality with KD, and to investigate the possible associations with clinical manifestations of the disease.

METHODSFour polymorphism sites of TNF-alpha and IL-10 gene promoter regions from 96 children with KD were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). One hundred and sixty age-matched normal children of the Han nationality were used as control. All patients accepted Doppler echocardiography examination in order to differentiate coronary artery lesions.

RESULTSThere was significant difference in allele frequencies of -308 (A/G) site of the TNF-alpha gene between children of the Han nationality and those of Japanese and Caucasian in America. There were significant differences in the allele frequencies of -1082 (G/A), -819 (C/T) and -592 (A/C) of IL-10 gene between children of the Han nationality and their British Counterparts (P < 0.01). There was no significant difference in allele frequencies of -308 (A/G) site of TNF-alpha gene between children with KD and normal controls. There was no significant difference in the haplotypes and the allele frequencies of the above three sites of IL-10 between the two groups. However, when clinical features were examined, the genotype frequency of TNF-alpha-308A was significantly higher in IVIG-resistant KD patients than that of TNF-alpha-308G genotype (67% vs 5%, chi(c)(2) = 90.48, P < 0.01). The genotype of TNF-alpha-308A was closely associated with IVIG-resistant KD (P < 0.01, relative risk 42.25, 95% confidence interval 15.81-112.88). The haplotype frequency of IL-10 -1082A/-819T/-592A was also higher in patients with coronary artery lesion (CAL) caused by KD than those of Non-ATA haplotype (52% vs 20%, chi(2) = 18.36, P < 0.01). The haplotypes of IL-10 -1082A/-819T/-592A was significantly associated with CAL caused by KD (P < 0.01, relative risk 4.26, 95% confidence interval 2.20-8.25).

CONCLUSIONThe genotype of TNF-alpha-308A is one of the important factors that probably influence the therapeutic effect of KD. The haplotypes (-1082/-819/-592) of IL-10 gene promoter might be related to the pathogenesis of coronary artery complication of KD and -1082A/-819T/-592A haplotypes might be regarded as a genetic marker of risk factor for coronary artery lesion in KD.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Interleukin-10 ; genetics ; Male ; Mucocutaneous Lymph Node Syndrome ; genetics ; pathology ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Promoter Regions, Genetic ; genetics ; Tumor Necrosis Factor-alpha ; genetics

BACKGROUNDTrichophyton rubrum (T. rubrum) represents the most important agent of dermatophytosis in humans. T. rubrum infection causes slight inflammation, and tends to be chronic and recurrent. It is suggested that it may result from the failure of epithelial cells to recognize T. rubrum effectively and initiate effective immune responses. The C-type lectin receptors (CLR) and toll-like receptors (TLR) are the two major pattern recognition receptors (PRRs) that recognize fungal components. Therefore, the purpose of the study was to analyze the expression of those PRRs and the cytokines in HaCaT cells stimulated with heat-inactivated T. rubrum conidia and hyphae, respectively.

METHODSHaCaT cells were unstimulated or stimulated with heat-inactivated T. rubrum conidia and hyphae (1×10(6) and 1.5×10(5) colony-forming unit (CFU) in 2 ml medium, respectively) for 6, 12 and 24 hours. The mRNA expression of PRRs involved in recognizing fungal pathogen-associated molecular patterns (PAMPs) and signaling molecules were measured by quantitative reverse transcription polymerase chain reaction (RT-PCR). Meanwhile, surface toll-like receptor (TLR) 2, TLR4 and Dectin-1 were analyzed by fluorescence-activated cell sorter (FACS) 24 hours after treatment. The cytokines were detected in cell culture supernatants of HaCaT cells in 12 and 24 hours after treatment.

RESULTSHaCaT cells constitutively expressed mRNA of membrane-bound TLR1, 2, 4 and 6, Dectin1 and DC-SIGN, but not Dectin-2 or Mincle. Heat-killed T. rubrum did not significantly upregulate gene transcriptions of the PRRs of HaCaT cells. Heat-inactivated T. rubrum conidia significantly reduced the surface expression of TLR2 and Dectin-1, and suppressed the secretions of interferon-inducible protein-10 (IP-10) and monocyte chemotactic protein-1 (MCP-1) of HaCaT cells, while heat-killed T. rubrum hyphae significantly induced the secretions of IP-10 and MCP-1.

CONCLUSIONThe cell-wall antigens of T. rubrum fail to activate transcriptional expression of PRRs and induce a lower immune response of HaCaT cells by limited cytokines secretion.

Cells, Cultured ; Cytokines ; biosynthesis ; Humans ; Keratinocytes ; immunology ; Lectins, C-Type ; genetics ; physiology ; RNA, Messenger ; analysis ; Receptors, Pattern Recognition ; genetics ; physiology ; Toll-Like Receptor 2 ; physiology ; Trichophyton ; immunology

BACKGROUNDTrichophyton rubrum is superficial fungi characteristically confined to dead keratinized tissues. These observations suggest that the soluble components released by the fungus could influence the host immune response in a cell in contact-free manner. Therefore, this research aimed to analyze whether the culture supernatant derived from T. rubrum grown in the nail medium could elicit the immune response of keratinocyte effectively.

METHODSThe culture supernatants of two strains (T1a, T XHB ) were compared for the β-glucan concentrations and their capacity to impact the innate immunity of keratinocytes. The β-glucan concentrations in the supernatants were determined with the fungal G-test kit and protein concentrations with bicinchoninic acid protein quantitative method, then HaCaT was stimulated with different concentrations of culture supernatants by adopting morphological method to select a suitable dosage. Expressions of host defense genes were assessed by quantitative polymerase chain reaction after the HaCaT was stimulated with the culture supernatants. Data were analyzed with one-way analysis of variance, followed by the least significant difference test.

RESULTSThe T. rubrum strains (T1a and T XHB ) released β-glucan of 87.530 ± 37.581 pg/ml and 15.747 ± 6.453 pg/ml, respectively into the media. The messenger RNA (mRNA) expressions of toll-like receptor-2 (TLR2), TLR4, and CARD9 were moderately up-regulated in HaCaT within 6-h applications of both supernatants. HaCaT cells were more responsive to T1a than T XHB . The slight increase of dendritic cells-specific intercellular adhesion molecule 3-grabbing nonintegrin expression was faster and stronger, induced by T1a supernatant than T XHB . The moderate decreases of RNase 7, the slight up-regulations of Dectin-1 and interleukin-8 at the mRNA level were detected only in response to T1a rather than T XHB . After a long-time contact, all the elevated defense genes decreased after 24 h.

CONCLUSIONThe culture supernatant of T. rubrum could directly and transiently activate the innate immune response of keratinocytes.

Cell Line, Tumor ; Culture Media, Conditioned ; pharmacology ; Humans ; Immunity, Innate ; drug effects ; Keratinocytes ; drug effects ; metabolism ; Trichophyton ; metabolism ; beta-Glucans ; metabolism