Objective To explore the role of plasminogen activator inhibitor-1(PAI-1)in development of progressive fibrosis via the inhibition of extracellular matrix degradation,and to reveal the contributive role of PAI-1 in muscular dystrophy(MD).Methods Expression and cellular localization of PAI-1 protein were examined in frozen muscle specimens obtained via biopsy from 5 patients with duchenne muscular dystrophy(DMD),3 patients with becker muscular dystrophy(BMD),9 patients with congenital muscular dystrophy(CMD) and 4 cases with normal muscle by immunohistochemistry,double immunofluorescence and Western-blot analysis.Results PAI-1 was positive only in vascular endothelial cells of normal muscle.Both immunohistochemistry and Western-blot analysis showed that PAI-1 expression distinctly increased in most dystrophic muscles of MD than that in normal muscles.Double immunolabeling revealed that PAI-1 strongly expressed in cytoplasm and nuclei of regenerating muscle fibers,macrophages,macrophage infiltrating necrotic fibers.Some activated fibroblasts in endomysium and perimysium of DMD and CMD muscles were positive for PAI-1.Conclusions The functional consequence of overexpression of PAI-1 in dystrophic muscles is unknown but the elevated local expression of PAI-1 in diseased muscles of MD and their distinct distribution pattern provide evidence that PAI-1 participate in pathogenesis of MD.

Objective To evaluate the impact of the project diverting the Yangtze River water to the Northern Jiangsu on transmission towards the north of Oncomelania hupensis snails, in order to provide the scientific basis for the South-North Water Diversion Project (east route). Methods The running situation of the water diversion project, the data on hydrology of the waterway and the distribution of snails and schistosomiasis were investigated with the methods of epidemiology、hydrology and field test. Results There was no significant correlation between the region of waterhead and water supply on the change of snail areas. There was no significant correlation between the snail areas of the region of water supply and the volume of water diversion, either. The snail habitat was stable at the Gaoyou segment along the Grand Canal. The snail habitat in the natural place of the north of 33?15′ north latitude on the east route has not been discovered yet. Conclusion There is no sign of transmission towards north on the snail habitat at the Gaoyou segment along the Grand Canal since the project was constructed in 1961. There is no evidence that the project leads to the spread of snails and schistosomiasis, either.

AIM:To explore the molecular mechanism of panaxadiol saponin(PDS)by observing Toll like receptor(TLR)2 and TLR9 mRNA expression induced by lipopolysaccharide(LPS).METHODS:Rats were divided into LPS,LPS+PDSL,LPS+PDSM and control group,respectively.Nitric oxide synthase(NOS)activity,nitric oxide(NO)content,LPO content,SOD activity and TLR2 and TLR9 mRNA expression were assayed 4 h after intravenous injection of LPS.RESULTS:NOS activity,NO content,LPO content of LPS+PDSL group and LPS+PDSM group were significantly lower than those in LPS group.TLR2 mRNA expression in the liver tissue of LPS+PDSL group and LPS+PDSM group was decreased compared with LPS group.CONCLUSION:PDS has a protective effect on liver tissues by triggering the down-regulation of TLR2 expression,reducing NOS activity,and NO content.

OBJECTIVEProgressive muscular dystrophy (PMD) is characterized by muscle fiber necrosis, regeneration, and endomysial fibrosis. Although absence of dystrophin and subsarcolemmic protein has been known as the cause of muscle fiber degeneration, pathogenesis of interstitial fibrosis is still unknown. The aim of this study was to investigate the role of connective tissue growth factor (CTGF) in PMD and its relationship with muscular fibrosis.

METHODSImmunological localization of CTGF was examined in frozen muscle specimens obtained via biopsy from 8 patients with Duchenne muscular dystrophy (DMD), 2 patients with Becker muscular dystrophy (BMD), 6 patients with congenital muscular dystrophy (CMD) and 6 cases with normal muscle by immunohistochemistry, double immunofluorescence and Western blot analysis.

RESULTSThe results of immunohistochemistry and double immunofluorescence showed that CTGF was positive only in vessels of normal muscle. Both immunohistochemistry and Western blot analysis showed that CTGF expression was distinctly increased in dystrophy muscles of PMD than that in normal muscles. In dystrophy muscle, marked immunostaining of CTGF was not only observed in vascular walls, but also strongly expressed in the cytoplasm and nuclei of regenerating muscle fibers, and also immunolocalized in the muscle fiber sarcolemma of non-regenerating fibers. Double labeling with antibodies against CTGF and CD68 demonstrated that CTGF was expressed in some macrophages and some macrophage infiltrated necrotic fibers. CTGF was strongly expressed in endomysial and perimysial connective tissues of dystrophy muscles of patients with DMD, CMD and FCMD. Double immunolabeling revealed that most activated fibroblasts in perimysium and endomysium were positive for CTGF, but not all of connective tissues were co-localized with CTGF. Older cases with FCMD showed poor or no expression of CTGF in advanced fibrosis.

CONCLUSIONCTGF may play a role in the pathogenetic process of muscular dystrophy, and CTGF may be important for muscle repair and fibrosis.

Adolescent ; Case-Control Studies ; Child ; Child, Preschool ; Connective Tissue Growth Factor ; metabolism ; Female ; Fibrosis ; Fluorescent Antibody Technique ; Humans ; Immunohistochemistry ; Infant ; Male ; Muscles ; metabolism ; pathology ; Muscular Dystrophies ; metabolism

OBJECTIVETo study serum concentration and mRNA expression of interleukin-13 (IL-13) in children with steroid-responsive nephrotic syndrome (SRNS) and the effect of methylprednisolone pulse therapy (MPT) on IL-13 expression.

METHODSTwenty-eight children with SRNS were enrolled in this study. Serum protein level of IL-13 was measured using ELISA and IL-13 mRNA expression in peripheral blood mononuclear cells (PBMC) was detected with RT-PCR before MPT, 2 and 5 days after MPT, and 2 weeks after disappearance of proteinuria following MPT. Twenty-four urinary protein was measured with the biuret assay. Twenty healthy children were used as controls.

RESULTSSerum IL-13 levels (38.48 +/- 13.01 pg/mL vs 5.18 +/- 2.71 pg/mL) and PBMC IL-13 mRNA expression (1.31 +/- 0.23 vs 0.36 +/- 0.07) before MPT in SRNS patients were significantly higher than in the controls. After 5 days of MPT and 2 weeks after disappearance of proteinuria following MPT, serum IL-13 levels (15.33 +/- 7.81 and 5.35 +/- 2.12 pg/mL respectively) and PBMC IL-13 mRNA expression (0.89 +/- 0.26 and 0.33 +/- 0.08 respectively) were significantly reduced (P < 0.01). Serum IL-13 levels and PBMC IL-13 mRNA expression in SRNS patients 2 weeks after disappearance of proteinuria following MPT were reduced to control levels, but remained at a higher level than controls 5 days after MPT. A positive correlation was found between serum levels of IL-13 and 24-hour urinary protein in SRNS patients before (r=0.75, P < 0.01) and after 2 and 5 days of MPT (r=0.68, r=0.71 respectively; P < 0.05).

CONCLUSIONSSerum IL-13 levels and PBMC IL-13 mRNA expression in children with SRNS increase. MPT can inhibit the expression of protein and mRNA of IL-13 in these patients.

Adolescent ; Child ; Female ; Humans ; Interleukin-13 ; blood ; genetics ; Male ; Methylprednisolone ; administration & dosage ; Nephrotic Syndrome ; blood ; drug therapy ; Proteinuria ; drug therapy ; RNA, Messenger ; analysis

OBJECTIVETo investigate the feasibility of multiple displacement amplification (MDA) to apply in the non-invasive prenatal genetic diagnosis of Duchenne muscular dystrophy (DMD).

METHODSMaternal blood was obtained from 20 pregnant women at 7 to 25 weeks of gestation. After the discontinuous density gradient centrifugation with Percoll, the fetal nucleated red blood cells (NRBCs) were stained with Kleihauer test. All positive NRBCs were collected by micromanipulator and then performed with MDA. Sex and short tandern repeat (STR) analysis were determind from a small aliquot of the reaction. The origin of NRBCs was verified and prenatal diagnosis of DMD was made at the same time.

RESULTSThe product length of MDA was >15 kb, while primer extension preamplification (PEP) is only about 1 kb. We completed non-invasive prenatal genetic diagnosis of 6 fetus at high risk of DMD using MDA. The results were all coincident with amniotic fluid control.

CONCLUSIONThe MDA method which provides a highly uniform representation across the genome, representing the entire genome with minimal amplification bias, shows good application prospects.

Erythroblasts ; metabolism ; Feasibility Studies ; Female ; Fetal Diseases ; blood ; diagnosis ; genetics ; Humans ; Muscular Dystrophy, Duchenne ; blood ; diagnosis ; genetics ; Polymerase Chain Reaction ; methods ; Pregnancy ; Prenatal Diagnosis ; methods

OBJECTIVETo investigate the clinical efficacy of Shengjing prescription for oligoasthenozoospermia and its action mechanism.

METHODSWe equally assigned 120 patients with oligoasthenozoospermia to receive Shengjing prescription (treatment group) and vitamin E (control group), respectively, for 12 weeks. Before and after the treatment, were obtained sperm concentration, sperm motility, the percentage of morphologically normal sperm, the levels of serum follicle-stimulating hormone (FSH), testosterone (T) and luteinizing hormone (LH), sperm DNA fragmentation index (DFI), the percentage of hypotonic swelling sperm, and the levels of seminal plasma elastase, x-glucosidase, fructose, zinc and acrosin.

RESULTSCompared with vitamin E, Shengling prescription significantly improved sperm concentration, motility and morphology (P < 0.01), decreased the serum FSH level, elevated the serum T level (P <0. 01) , reduced DFI and seminal plasma elastase, and increased the percentage of hypotonic swelling sperm as well as the levels of seminal plasma cx-glucosidase, fructose, zinc and acrosin.

CONCLUSIONShengjing prescription improves semen parameters of patients with oligoasthenozoospermia at multiple levels and through multiple channels.

Adult ; Asthenozoospermia ; drug therapy ; physiopathology ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Infertility, Male ; Male ; Phytotherapy ; Sperm Count ; Sperm Motility ; Treatment Outcome ; Vitamin E ; therapeutic use

BACKGROUNDT-cell receptor (TCR) plays an important role in the development of autoimmune diseases. Recently, it was reported that immunization of animals with TCR peptide derived from the pathogenic cells could prevent autoimmune diseases. The aim of this study was to investigate whether vaccination with a synthetic peptide from the hypervariable region of TCR V(beta) 8.3, an experimental autoimmune uveoretinitis (EAU)-associated gene, was able to prevent the disease.

METHODSEAU was induced in Lewis rats by immunization with IRBP R16 peptide emulsified in complete Freund's adjuvant (CFA). The clinical and histological appearances were scored. Delayed type hypersensitivity (DTH) and lymphocyte proliferation were detected. Cytokine levels of aqueous humour, supernatants of cells from spleen and draining lymph nodes were measured by enzyme linked immunosorbent assay (ELISA). Gene expression of TCR V(beta) 8.3 on CD(4)(+) T cells was examined by real time quantitative polymerase chain reaction (PCR).

RESULTSAfter vaccination, the intraocular inflammation was significantly mitigated, antigen specific DTH and lymphocyte proliferation responses were suppressed, interleukin (IL)-2 in aqueous humour, interferon (IFN)-gamma and IL-2 produced by the spleen and draining lymph node cells were significantly decreased, whereas the production of IL-4 and IL-10 were increased. The response of draining lymph node cells to TCR V(beta) 8.3 peptide was enhanced after vaccination. Inoculation with CFA alone did not affect the severity of EAU and the above parameters. The suppression of EAU was much stronger in the group of four fold inoculations than the group of two fold inoculations. The expression of TCR V(beta) 8.3 gene was significantly reduced in the group of fourfold inoculations.

CONCLUSIONVaccination with the synthetic TCR V(beta) 8.3 peptide could remarkably inhibit the development of EAU.

Animals ; Autoimmune Diseases ; prevention & control ; Cytokines ; biosynthesis ; Female ; Genes, T-Cell Receptor beta ; Rats ; Rats, Inbred Lew ; Receptors, Antigen, T-Cell, alpha-beta ; immunology ; Retinitis ; prevention & control ; Retinol-Binding Proteins ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Uveitis ; prevention & control ; Vaccination

Objective Tissue inhibitor of metalloproteinase-1 (TIMP-1) is a multifunctional protein that has thc capacity to modify cellular activities and to modulate matrix turnover. This paper revealed the contributive role of TIMP-1 in progressive muscular dystrophy (PMD). Methods We examined the expression and cellular localization of TIMP-1 protein using biopsied frozen muscle from patients with Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD), congenital muscular dystrophy (CMD) by immunohistochemistry, double immunofluorescence and Western blot analysis. Results The results of immunohistochemistry and double immunofluorescence showed that TIMP-1 was positive only in vascular endothelial cells of normal muscles. Immunohistochemistry and Western blot analysis showed that the staining intensity was distinctly increased in some dystrophic muscles of PMD for TIMP-1. Double immunofluorescence revealed that TIMP-1 strongly expressed in the regenerating muscle fibers, macrophages and macrophage infiltrating necrotic fibers. Some activated fibroblasts in endomysium and perimysium of DMD and CMD muscles were also positive for TIMP1. Conclusion The functional consequence of overexpression of TIMP-1 in the dystrophic muscles is unknown, but the elevated local expression of TIMP-1 in diseased muscles of PMD and their distinct distribution pattern provide evidence that TIMP-1 may participate in the pathogenesis of PMD.

OBJECTIVETo study the newborn's Apgar score in 'one minute' and relevant factors.

METHODSOne year inpatient woman from a Maternal and Child Health Hospital of Anhui province were selected by cluster sampling method and newborn asphyxia situation was investigated using Apgar score and self-designed questionnaire.

RESULTSThe Apgar score in 'one minute' which marking 8 to 10, 4 to 7 and 0 to 3 were found in 1875 (73.78%), 426 (16.77%) and 240 infants (9.45%) respectively. The average Apgar score in 'one minute' and five minutes were (7.69 +/- 2.27) and (9.01 +/- 1.89) respectively, The Apgar score in 'one minute' was significantly correlated with that of five minutes (Pearson coefficient correlation r = 0.841, P = 0.00). Ordinal regression analysis revealed that parturient age (OR = 1.04), being farmer (OR = 2.22), parity (OR = 1.26), assistant vaginal delivery (OR = 4.93), caesarean section (OR = 1.95), pregnancy-induced hypertension syndrome (OR = 1.42), albuminuria in gestational period (OR = 1.44), newborn being male (OR = 1.23), low birth weight (OR = 2.94), inborn abnormality (OR = 12.12), premature birth (OR = 1.22) and complications of delivery (OR = 5.04) were risk factors while the number of years under study (OR = 0.91), prenatal check-up (OR = 0.48), body length of newborn infant (OR = 0.88) and single birth (OR = 0.57) were protective factors.

CONCLUSIONApgar score in 'one minute' of newborn infant was affected by several factors as stated above. Health care program in earlier period toward community parturient should be strengthened in order to discover and control high risk factors of duration of pregnancy in earlier period.

Apgar Score ; Asphyxia Neonatorum ; epidemiology ; Epidemiologic Studies ; Female ; Humans ; Infant, Newborn ; Male ; Pregnancy ; Risk Factors ; Time Factors