OBJECTIVETo observe the effect of Astragalus-Angelica Mixture (AAM) on osteopontin (OPN) expression in rats with chronic nephrosclerosis.

METHODSChronic nephrosclerosis model rats induced by repeated intraperitoneal injection of puromycin were randomly divided into the model group, AAM group and Irbesartan (an antagonist of angiotensin) group. The experimental course lasted 12 weeks. Blood and urine samples were examined by biochemical method. Kidney tissue was taken for pathological stain and immunohistochemical method and was applied to examine OPN expression, mononuclear macrophage, laminin in extracellular matrix and decorin expressions.

RESULTSAAM showed the effects of decreasing urinary protein and improving renal function similar to that of Irbesartan. It also could alleviate the pathological damage of kidney tissue, especially in decreasing renal tubular mesenchymal damage index. The accumulation of decorin and laminin in the mesenchymal extracellular matrix significantly decreased. Renal tubular OPN expression and mesenchymal infiltration of mononuclear macrophage decreased significantly and in a positive correlated manner (r = 0.885, P < 0.01).

CONCLUSIONAAM has similar renal protective action to that of Irbesartan, this action may be related to the inhibition of up-regulated OPN expression.

Animals ; Astragalus Plant ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Male ; Nephrosclerosis ; chemically induced ; metabolism ; Osteopontin ; Phytotherapy ; Puromycin ; Rats ; Rats, Sprague-Dawley ; Sialoglycoproteins ; biosynthesis

The present study was aimed to analyze the immunogenicity of recombinant human zona pellucida-3 peptides (r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348)) expressed in E. coli through immunizing rabbits, and to evaluate the efficacy of their polyclonal antisera against r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) to inhibit in vitro human sperm-egg binding respectively. Male New Zealand rabbits were immunized using r-huZP3a(22 approximately 176) or r-huZP3b(177 approximately 348) as antigen respectively, which was purified through an improved method of preparative gel polyacryulamide gel electrophoresis. The antibody response level of r-huZP3a(22 approximately 176) or r-huZP3b(177 approximately 348) immunogen in rabbits was determined by ELISA using mouse ZP3-5 (amino acid sequence(137 approximately 150) being completely conserved with huZP3(138 approximately 151) sequence) and specific huZP3-14 (amino acid sequence(327 approximately 340)) synthetic peptides as coating antigens respectively. The immunoreactivity and specificity of the anti-r-huZP3a(22 approximately 176) and anti-r-huZP3b(177 approximately 348) antisera with each r-huZP3 peptides, were tested by immunoblot and immunohistochemistry (using native huZP and human ovary section) respectively. A competitive hemizona assay (HZA) was used to evaluate the efficacy of the antisera against r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) to inhibit in vitro human sperm-egg binding. Both r-huZP3 peptides were able to induce higher antibody titers in rabbits. Each antiserum could specifically recognize or bind to each target r-huZP3 peptide expressed in E. coli and native human ZP in vitro. The antisera also inhibited sperm-egg binding in the HZA. These results show that r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) are of strong immunogenicity. They can be used to develop a kit for detecting whether there are autoantibodies to zona pellucida in unexplained infertile women, and their antisera might be useful tools for determining minimal B-cell epitope sequences of several known huZP3 epitope peptides.
Animals ; Egg Proteins ; biosynthesis ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Female ; Humans ; Immune Sera ; immunology ; Immunization ; Male ; Membrane Glycoproteins ; biosynthesis ; genetics ; immunology ; Rabbits ; Receptors, Cell Surface ; biosynthesis ; genetics ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Sperm-Ovum Interactions ; immunology ; Zona Pellucida Glycoproteins

To characterize the genomic sequence and arrangement of WU polyomavirus (WU virus) identified in clinical specimens collected from children with acute respiratory infections in Beijing, China, the sequences of capsid proteins VP1, VP2, and the large tumor antigen (LTAg), as well as the 5'-terminal sequence of WU virus, were amplified from the clinical specimen with ID number of BJF5276 which was determined as WU virus positive by PCR amplification. The PCR amplicons were sequenced, and genomic sequence analysis was performed by using the software DNAStar. In addition, VP2 coding-region sequences were amplified from other 21 clinical specimens identified as WU virus positive to investigate the gene diversity of WU virus. The genomic sequence of WU virus BJF5276 with accession number of HQ218321 in GenBank was 5,229 base pairs in length with 3 major coding domain sequences (CDS) sited on one strand coding for capsid proteins VP2, VP3 and VP1, and two CDS sited on the complementary strand coding for small tumor antigen (STAg) and LTAg; These 22 VP2 CDS sequences including 5 sequences submitted to GenBank were compared with 64 corresponding sequences downloaded from GenBank by MegAlign of DNAStar software, indicated that these sequences coming from children in Beijing shared high homology (over 98.8%) with those from GenBank. Phylogenetic analysis of these VP2 CDS by using Neighbor-joining (NJ) analyses with 2,000 bootstraps (Mega 4.0) showed that 20 sequences out of 22 belonged to clade Ia, and other 2 of them belonged to clade III, including 1 clustered in IIIa and 1 in a novel cluster proposed as IIIc. In conclusion, the genomic sequence of WU polyomavirus detected from clinical specimens from children in Beijing is closely related to other WU polyomaviruses in the feature of genomic coding region arrangement. Overall variation of VP2 CDS was very low, and there were different clades circulating in Beijing with a dominant clade Ia, which is different from dominated Ib circulating in other parts of the world reported previously, and a novel clade IIIc was proposed.
Acute Disease ; Child, Preschool ; China ; Female ; Genome, Viral ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny ; Polyomavirus ; classification ; genetics ; isolation & purification ; Respiratory Tract Infections ; virology ; Viral Proteins ; genetics

In order to solve the problem of selection and in vivo delivery problem in siRNA treatment, hepatitis B virus (HBV) HBx gene which could be targeted by siRNA was studied. The siRNA expression plasmid which specific inhibits HBx expression was obtained by in vitro selection via a dual-luciferase plasmid including HBx-Fluc fusion protein expression domain. The selected siRNA expression plasmid was then encapsulated in PEG-modified cationic liposome, which was devoted into pharmacodynamic studies at both cellular and animal level. The results illustrated that the cationic liposome which encapsulated siRNA expression plasmid could effectively inhibit HBx gene expression both in vitro and in vivo.
Cations ; Gene Expression Regulation, Viral ; drug effects ; Hepatitis B virus ; genetics ; Liposomes ; chemistry ; Plasmids ; RNA, Small Interfering ; chemistry ; Trans-Activators ; genetics ; metabolism

Objective To determine the urinary iodine level of people in Yunyang and Bishan County of Chongqing and explore into its influencing factors. Methods Using multistage cluster stratified simple random sample method, Yunyang and Bishan County were chosen as research spots, then thirty children aged 8-10 in each 3 primary school of the 2 counties were selected using stratified randomization sampling method to inspected their urine and household salt for iodine and the iodine content in drinking water. Results Five hundred and seventy-one urine samples were inspected and the urinary iodine median was 261.47 μg/L. 5.78% (33/571) and 37.48%(214/571) of samples had an urinary iodine median less than 100 μg/L and more than 300 μg/L. The urinary iodine median of Yunyang County was higher than that of Bishan (H = 7.42, P < 0.01). The iodine salt coverage rate, the qualified rate and edible qualified iodine salt rate respectively were 99.64%(554/556), 94.22% (522/554) and 93.88% (522/556) in 556 samples of family table salt. Eighty-seven samples of drinking water were inspected, resulting an averaged iodine content of 8.81 and 2.97 μg/L, respectively in the 2 counties. Conclusions The 2 counties are all the area of iodine deficiency. The urinary iodine level, although meeting the demand of eliminating iodine deficiency diseases, is a little bit higher given that iodized salt of present doage has been taken for a long time. The content of iodized salt should be adjusted accordingly.

Objective To explore the application of entire information management system in nursing of patients with high-risk pressure ulcers and to evaluate its influence on the high risk of pressure ulcers prevention management.Methods Established core group of pressure ulcers management to provide professional guidance to high-risk pressure ulcers patients nursing in whole hospital, structured information management module, improved high-risk pressure ulcers report, added statistical form for it, implemented intelligent record as well as automatic statistics, improved and implemented nursing management work flow for high risk pressure ulcers patients, strengthen supervise to the practical to pressure ulcers prevention, and dynamically managed the whole process from reporting to lapsing of high risk patients by information system.Results After the implementation of entire information management system, pressure ulcers incidence of high risk patients in hospital was significantly decreased compared with that of before(18.98%vs 32.64%;χ2 =10.032,P<0.01).The report accuracy of pressure ulcers was improved than that before (98.15% vs 91.71%;χ2 =9.084,P<0.01).The time for nursing administration department making statistics for pressure ulcers case quarterly was decreased from 6 hours to 20 minutes.Conclusions Adopting the entire information management system of pressure ulcers can decrease the pressure ulcers morbidity of high risk patients, increase the accuracy of report for high-risk pressure ulcer, improve work efficiency of statistical analysis to high-risk pressure ulcers, and improve the work for prevention and control of pressure ulcers.

Objective To assess the therapeutic effect and safety ofpramipexole combined with Madopar and Madopar alone in the treatment of Parkinson's disease (PD). Methods This randomized, controlled open-label trial involved 70 PD patients who were randomly assigned to receive pramipexole combined with Madopar (n=35) or Madopar alone (n=35) for 12 consecutive weeks. The therapeutic effect was assessed primarily by the Unified Parkinson's Disease Rating Scale (UPDRS). The main effect was motor symptoms of UPDRS Ⅲ and activities of daily living of UPDRS Ⅱ. The secondary effect was the changes relative to the baseline levels in the consciousness, behavior, and emotion of UPDRS Ⅰ, the complications of UPDRS Ⅳ and daily Madopar dosage. The safety of the drugs was evaluated according to the adverse reactions. Results Compared to the baseline levels, 12 weeks of treatment with pramipexole combined with Madopar resulted in significantly greater reduction in the total scores ofUPDRS Ⅲ than Madopar alone (11.40 vs 9.26, P<0.05), but the reduction in the total DOI: 10.3760/cma.j .issn. 1671.8925.2009.07.010scores of UPDRS Ⅱ and UPDRS Ⅰ was comparable between the two treatments (4.57 vs 4.50 and 0.66 vs 1.14, respectively, P>0.05). The combined treatment reduced the total score of UPDRS Ⅳ by 0.22 whereas Madopar alone increased the score by 0.06, showing significant difference between the two treatments (P<0.05). The daily Madopar dosage in the combined treatment group was decrease by 163.57 rag, but that in Medopar group increased by 8.57 rag (P<0.05). The frequencies of wearing-off, symptom fluctuation and movement disorder were significantly lower in combined treatment group than in Madopar group. Obvious wearing-off, symptom fluctuation and movement disorder occurred in Madopar group, which were not noted in the combined treatment group but two patients developed sudden sleep, one reported drowsiness, and another exhibited orthostatic hypotension. Conclusion Pramipexole combined with Madopar results in better improvement of the motor symptoms than Madopar alone in PD patients, but they show similar effect on the activities of daily living and consciousness, behavioral and emotional changes. Pramipexole can significantly decrease the daily dose of Madopar and help reduce the complications, suggesting its safety and effectiveness in the treatment of PD, but its adverse effects should be given due attention in clinical application.

Objective To investigate the protective mechanism of insulin on paraquat-induced PC12 cells. Methods Based on the Part (insulin can protect the paraquat-induced PC12 cells), immunoprecipitation and Western blotting were employed to observe the protein expressions of INR Tyr~(1162/1163) and AktSer~(473) in the group of normal PC12 cells, group of insulin interfered PC12 cells, group of PQ-induced PC12 cells and group of PQ combined with insulin interfered PC12 cells, respectively. Results Group of insulin interfered PCI2 cells and group of PQ combined with insulin interfered PC12 cells showed expression of phosphorylation protein INR Tyr~(1162/1163), while the other groups without insulin intervention did not show the expression of that. The expression of phosphorylation protein INR Tyr~(1162/1163) in the group of insulin interfered PC12 cells was higher than that in the group of PQ combined with insulin interfered PC12 cells, but statistical analysis showed no significant difference between the 2 groups (P>0.05). Meanwhile, the expression of phosphorylation protein AktSer~(473) in the group of insulin interfered PC 12 cells was obviously higher than that in the group of PQ combined with insulin interfered PC12 cells (P<0.05). Conclusion The increased expression of phosphorylation protein INR Tyr~(1162/1163) and AktSer~(473) might be one of the clues in proving that insulin may have the protective effect on PC12 cells.

Amino Acid Sequence ; Base Sequence ; Genes, Bacterial ; Humans ; Molecular Sequence Data ; Polymerase Chain Reaction ; Staphylococcus epidermidis ; enzymology ; genetics ; beta-Lactamases ; chemistry ; genetics