Alleles ; Gene Amplification ; Genome ; Genotype ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; Humans ; Polymerase Chain Reaction ; methods

Objective To explore the effect of ACE-inhibitor perindopril on the expression of scavenger receptor A (SR-A) gene in the kidney of diabetic rats.Methods Diabetes were induced in male Sprague-Dawley rats by peritoneal injection with streptozotocin (60mg/kg).The rats were then random di vided into normal control group, diabetes group and ACEI treatment group [4mg/(kg·d) for 24 weeks].Blood glucose concentration and 24h urinary albumin excretion were determined.The renal morphological change was observed.Immunohistochemistry was used to analyze CD68 positive macrophages,and the Mrna of SR-A in renal tissue was detected by quantitative real-time PCR.Results Compared with normal control group,blood glucose concentration,24h urinary albumin excretion and the number of CD68 positive macrophages were significantly increased [(5.3 ± 0.6) mmol/L vs (26.7 ± 3.3) mmol/L;(2.7 ± 1.3) mg/24h vs (26.7 ± 1.8)mg/24h;(0.77 ±0.24)/gcs vs (2.55 ±0.46)/gcs;(6.13 ±0.50)/HPF vs (11.9 ±2.12)/HPF;P ＜0.05],and the expression of SR-A Mrna were significantly up-regulated in diabetes group [ (5.6 ± 1.2 vs 1.5 ±0.2),P ＜0.05].After intervention with ACE-inhibitor,the up-regulations of the above mentioned parameters,except blood glucose concentration,were all significantly inhibited [ (3.6 ±1.4)mg/24h;(1.03±0.37)/gcs;(8.28±1.19)/HPF;3.4±0.7;P ＜0.05].Conclusion ACE-inhibitor might have renoprotective effects of diabetic nephropathy,it probably was associated with inhibiting the expression of SR-A gene.

BackgroundRetinitis pigmentosa (RP)is a group of progressive monogenic inheritance disease.Seldom epidemiology is performed to summarize the varied clinical phenotypes,especially some sporadic cases with untypical genetic history.ObjectiveThe aim of this survey was to investigate the clinical epidemiological characteristics and phenotype of sporadic RP.MethodsA prospective cohort study was designed.A survey of a series of clinically diagnosed sporadic primary RP patients was conducted at the Southwest Eye Hospital from July 2010 to November 2011.A total of 130 patients that matched the inclusion criteria were enrolled in this survey.Clinical ocular examinations and questionnaire surveys were given,including ophthalmoscopic examination,best corrective visual acuity( BCVA ),perimetry and Ganzfield electroretinogram (ERG)and color fundus photo.RP with different phenotypes were classified. ResultsA total of 130 sporadic RP patients were collected in this survey.Of them,66 were male and 64 were female with a mean age of (36.9±14.4) years.The average onset age of these subjects was (21.2±18.4) years.Seven (5.38％) patients had consanguineous marriage history,and 13 ( 10.00％ )patients had systemic disease.Forty-four (33.85％) patients had outdoor jobs,and 86 (66.15％ ) worked indoor.Eighty-nine patients had typical RP ( 68.5％ ),and the number of patients that developed central RP and sine pigmento RP were 16 ( 12.3％ ) and 16( 12.3％ ),respectively.An absence of a- and b-waves in full-field ERG wasdetected in 99 (76.15％ ) cases.The longest duration of night blindness was identified in typical RP patients and the lowest BCVA in central RP patients.ConclusionsThe age at first onset is early in sporadic RP.There are wide variations in different types of RP,but the ERG outcome is specific for all RP types.

Adult ; Chromosome Aberrations ; Chromosomes, Human, Pair 21 ; Down Syndrome ; diagnosis ; Female ; Fertilization in Vitro ; Humans ; In Situ Hybridization, Fluorescence ; Maternal Age ; Pregnancy ; Pregnancy, High-Risk ; Preimplantation Diagnosis

Objective:To observe the clinical effect of lower-reinforcing and upper-reducing acupuncture method for hyperplasia of mammary gland (HMG) and its influence on estradiol (E2) and progesterone (P).Methods:A randomized,single-blinded and controlled trial was conducted.A total of 124 cases conforming to the inclusion criteria were randomized by random number table into a treatment group and a control group,with 62 cases in each group.Patients in both groups received acupuncture therapy at the same acupoints,while patients in the treatment group received lower-reinforcing and upper-reducing method,and patients in the control group received even reinforcing-reducing manipulation.The treatment started around 10 d before menstruation and was conducted every other day.Patients received 5 treatments in each menstruation cycle for consecutive 3 cycles.The levels of E2,P and E2/P and clinical efficacy were measured before and after treatment.Results:After treatment,the breast lump size,pain intensity and concomitant symptoms score in both groups were substantially lower than those before treatment,showing statistical significances (all P＜0.01),and the improvement in the treatment group was higher than that in the control group,and the between-group comparisons showed statistical significances (all P＜0.01).After treatment,the overall effective rate was 91.9％ in the treatment group,higher than 72.6％ in the control group,and the between-group comparison showed a statistical significance (P＜0.01).After treatment,levels of E2,P and E2/P value showed no statistical significance when compared with those before treatment in the two groups (all P＞0.05).Conclusion:Lower-reinforcing and upper-reducing acupuncture method can effectively alleviate clinical symptoms and signs in HMG patients,and produce a better effect than even reinforcing-reducing manipulation.The majority of HMG patients' E2,P level and E2/P value were not beyond the normal ranges;therefore,acupuncture showed no substantial influence on E2 and P levels and E2/P value.

Background::Acute-on-chronic liver failure (ACLF) is a severe liver disease with complex pathogenesis. Clinical hypoglycemia is common in patients with ACLF and often predicts a worse prognosis. Accumulating evidence suggests that glucose metabolic disturbance, especially gluconeogenesis dysfunction, plays a critical role in the disease progression of ACLF. Lon protease-1 (LONP1) is a novel mediator of energy and glucose metabolism. However, whether gluconeogenesis is a potential mechanism through which LONP1 modulates ACLF remains unknown.Methods::In this study, we collected liver tissues from ACLF patients, established an ACLF mouse model with carbon tetrachloride (CCl 4), lipopolysaccharide (LPS), and D-galactose (D-gal), and constructed an in vitro hypoxia and hyperammonemia-triggered hepatocyte injury model. LONP1 overexpression and knockdown adenovirus were used to assess the protective effect of LONP1 on liver injury and gluconeogenesis regulation. Liver histopathology, biochemical index, mitochondrial morphology, cell viability and apoptosis, and the expression and activity of key gluconeogenic enzymes were detected to explore the underlying protective mechanisms of LONP1 in ACLF. Results::We found that LONP1 and the expressions of gluconeogenic enzymes were downregulated in clinical ACLF liver tissues. Furthermore, LONP1 overexpression remarkably attenuated liver injury, which was characterized by improved liver histopathological lesions and decreased serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in ACLF mice. Moreover, mitochondrial morphology was improved upon overexpression of LONP1. Meanwhile, the expression and activity of the key gluconeogenic enzymes were restored by LONP1 overexpression. Similarly, the hepatoprotective effect was also observed in the hepatocyte injury model, as evidenced by improved cell viability, reduced cell apoptosis, and improved gluconeogenesis level and activity, while LONP1 knockdown worsened liver injury and gluconeogenesis disorders.Conclusion::We demonstrated that gluconeogenesis dysfunction exists in ACLF, and LONP1 could ameliorate liver injury and improve gluconeogenic dysfunction, which would provide a promising therapeutic target for patients with ACLF.

BACKGROUNDDengue is currently a significant global health problem but no vaccines are available against the four dengue serotypes virus infections. The development of safe and effective vaccines has been hampered by the requirement of conferring complete protection against all four dengue serotypes and the lack of a convenient animal model. Virus-like particles (VLPs) have emerged as a promising subunit vaccine candidate. One strategy of vaccine development is to produce a tetravalent dengue subunit vaccine by mixing recombinant VLPs, corresponding to all four dengue virus serotypes. Towards this end, this study aimed to establish a Pichia pastoris (P. pastoris) expression system for production of dengue virus type 1 (DENV-1) VLPs and evaluate the humoral and cellular immune response of this particle in mice.

METHODSA recombinant yeast P. pastoris clone containing prM and E genes of DENV-1 was constructed and DENV-1 VLPs expressed by this clone were analyzed by sucrose density gradient centrifugation, Western blotting, and transmission electron microscope. Groups of mice were immunized by these particles plus adjuvant formulations, then mice were tested by ELISA and neutralization assay for humoral immune response, and by lymphocyte proliferation and cytokine production assays for a cellular immune response.

RESULTSOur data demonstrated that recombinant DENV-1 VLPs consisting of prM and E protein were successfully expressed in the yeast P. pastoris. Sera of VLPs immunized mice were shown to contain a high-titer of antibodies and the neutralization assay suggested that those antibodies neutralized virus infection in vitro. Data from the T lymphocyte proliferation assay showed proliferation of T cell, and ELISA found elevated secretion levels of interferon IFN-γ and IL-4.

CONCLUSIONSP. pastoris-expressed DENV-1 VLPs can induce virus neutralizing antibodies and T cell responses in immunized mice. Using P. pastoris to produce VLPs offers a promising and economic strategy for dengue virus vaccine development.

Animals ; Antibodies, Neutralizing ; immunology ; Antibodies, Viral ; immunology ; Dengue Virus ; genetics ; immunology ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique ; Male ; Mice ; Mice, Inbred BALB C ; Pichia ; genetics ; metabolism ; T-Lymphocytes ; immunology

Non-progressive congenital myopathy is a group of muscle diseases occurring at birth or during teenage years. A number of new reports of congenital myopathy, such as homogeneous bodies myopathy, muscle quality control myopathy and type 1 fiber predominance have recently been reported, but they lack of sufficient quantity and constant clinico-pathologic manifestations. This paper reports two cases of congenital myopathy with type 1 fiber predominance confirmed by muscle biopsy. The clinical manifestations of the two children (a 4.5-year-old girl and an 11-year-old boy) included non-progressive symptoms of muscle weakness, skeletal deformities and other clinical features of congenital myopathy. The physical examinations showed a long face or figure and funnel chest or kyphosis/scoliosis, high palatal arch and wing-like shoulder. Serum levels of creatine kinase were normal but slightly elevated serum lactate dehydrogenase levels were noted in the two children. The skeletal muscle biopsy by ATPase staining showed that type 1 fibers accounted for more than 90% of the total number of muscle fibers. No other abnormal pathological changes, such as central cores, muscle tube and central nuclei, were found in the two children.
Diagnosis, Differential ; Female ; Humans ; Infant ; Male ; Muscle, Skeletal ; pathology ; Myopathies, Structural, Congenital ; diagnosis ; pathology ; therapy

Background Dengue is currently a significant global health problem but no vaccines are available against the four dengue serotypes virus infections.The development of safe and effective vaccines has been hampered by the requirement of conferring complete protection against all four dengue serotypes and the lack of a convenient animal model.Virus-like particles (VLPs) have emerged as a promising subunit vaccine candidate.One strategy of vaccine development is to produce a tetravalent dengue subunit vaccine by mixing recombinant VLPs,corresponding to all four dengue virus serotypes.Towards this end,this study aimed to establish a Pichia pastoris (P.pastoris) expression system for production of dengue virus type 1 (DENV-1) VLPs and evaluate the humoral and cellular immune response of this particle in mice.Methods A recombinant yeast P.pastoris clone containing prM and E genes of DENV-1 was constructed and DENV-1 VLPs expressed by this clone were analyzed by sucrose density gradient centrifugation,Western blotting,and transmission electron microscope.Groups of mice were immunized by these particles plus adjuvant formulations,then mice were tested by ELISA and neutralization assay for humoral immune response,and by lymphocyte proliferation and cytokine production assays for a cellular immune response.Results Our data demonstrated that recombinant DENV-1 VLPs consisting of prM and E protein were successfully expressed in the yeast P.pastoris.Sera of VLPs immunized mice were shown to contain a high-titer of antibodies and the neutralization assay suggested that those antibodies neutralized virus infection in vitro.Data from the T lymphocyte proliferation assay showed proliferation of T cell,and ELISA found elevated secretion levels of interferon IFN-y and IL-4.Conclusions P.pastoris-expressed DENV-1 VLPs can induce virus neutralizing antibodies and T cell responses in immunized mice.Using P.pastoris to produce VLPs offers a promising and economic strategy for dengue virus vaccine development.

Animals ; Bone Marrow ; drug effects ; Catechin ; pharmacology ; Fabaceae ; chemistry ; Female ; Gene Expression Regulation ; drug effects ; Granulocyte-Macrophage Colony-Stimulating Factor ; blood ; genetics ; Hematopoiesis ; drug effects ; Interleukin-6 ; blood ; genetics ; Male ; Mice ; Plants, Medicinal ; chemistry ; RNA, Messenger ; analysis