Calcinosis ; Female ; Humans ; Infant, Newborn ; Vascular Diseases ; pathology

Objective To observe the influence of taurine supplementation during early postnatal life on body weight and ultrastructure of islet ? cells in neonatal rats with low birth weight(LBW).Methods LBW neonatal rats were made by feeding 20%(C group) or 10%(R group) protein diet to fetal rats during gestation and lactation.Half of femal rats in group R were given a supplementation with 2.5% taurine drinking water(RT group) only during lactation,while other femal rats freely drunk.At postnatal day 1 and 21,the neonatal rats were weighted and their pancreas were removed.The ultrastructural changes of ? cells were observed by electron microscopy.Results At postnatal 21 days,the body weight of offsprings in group RT was significantly highter than that in group R(P=0.003);and the ultrastructure of ? cells in group RT got more improvement than that in group R.Conclusion Taurine supplementation can improve the growth-catch-up and the ultrastructure of islet ? cells of neonateal rats with LBW.

ObjectiveTo explore the effect of brain wave-biofeedback on attention deficit hyperactivity disorder.Methods29 children with attention deficit hyperactivity disorder used VBFB3000 Brain Wave-Biofeedback system to control the 4～8 Hz brain wave and activate the 12～16 Hz wave twice a week.Results84.6% children primarily with attention deficit became normal,as well as 100% with hyperactivity,91.6% with mixed appearing.ConclusionBrain Wave-Biofeedback is effective on any types of attention deficit hyperactivity disorder.

OBJECTIVETo investigate the effect of oligochitosan in promoting intestinal absorption of protoberberine alkaloids in extracts from Corydalis saxicola total alkaloids.

METHODThe in vitro single-pass intestinal perfusion model in rats was established to study the changes in absorption kinetic parameters of dehydrocavidine, berberine hydrochloride and palmatine chloride in C. saxicola total alkaloids after the addition of different concentrations oligochitosan and evaluate the effect of oligochitosan in promoting intestinal absorption of the drugs.

RESULTThe concentration of oligochitosan had different effects on the absorption rate constant (Ka) and apparent permeability coefficient (Peff) of the three active component in rat intestines. Ka and Peff in 0.5% oligochitosan group significantly increased, indicating a stronger effect in promoting the absorption.

CONCLUSIONOligochitosan has a certain effect in promoting the intestinal absorptions of protoberberine alkaloids in C. saxicola total alkaloids.

Animals ; Berberine Alkaloids ; administration & dosage ; pharmacokinetics ; Chitin ; administration & dosage ; analogs & derivatives ; Corydalis ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore efficacy enhancing and detoxification roles of Jiedu Quyu Zishen Recipe (JQZR) in treating systemic lupus erythematosus (SLE) by studying its effect on Toll like receptor 9 (TLR9) signal pathway of murine macrophage cells after JQZR stimulated CpG oligodeoxynucletide (CpG ODN).

METHODSMurine macrophage cells in vitro cultured were randomly divided into 4 groups, i.e., the blank serum group, the CpG ODN stimulus group, the CpG ODN + dexamethasone group, the CpG ODN + medicated serum group. Murine macrophage cells were collected after 24-h intervention. The expression of TLR9, myeloid differentiation factor 88 (MyD88), NF-KB, IFN-α mRNA were analyzed by RT-PCR. The expression of TLR9 and NF-κB protein were analyzed by Western blot. Changes of the NF-KB transcriptional activity were assayed by Dual-Luciferase reporter assay system.

RESULTSmRNA expressions of TLR9, MyD88, NF-κB, and IFN-α, protein expressions of TLR9 and NF-κB, and NF-κB transcriptional activities were enhanced, showing statistical difference when compared with those of the blank serum group (P <0. 05, P <0. 01). Compared with the CpG ODN stimulus group, mRNA expressions of MyD88, NF-κB, and IFN-α, the protein expression of NF-κB and the NF-κB transcriptional activities decreased in the CpG ODN + dexamethasone group with statistical difference (P <0. 01). Compared with the CpG ODN stimulus group, mRNA expressions of TLR9, MyD88, NF-κB, and IFN-α, protein expressions of TLR9 and NF-κB, and NF-κB transcriptional activities were decreased in CpG ODN+ medicated serum group with statistical difference (P <0. 01).

CONCLUSIONEfficacy enhancing and detoxification roles of JQZR in treatment of SLE might be realized through regulating TLR9 signal pathways.

Animals ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Macrophages ; metabolism ; Mice ; Myeloid Differentiation Factor 88 ; NF-kappa B ; RNA, Messenger ; Signal Transduction ; Toll-Like Receptor 9 ; metabolism

Twenty-five compounds of novel quinoxaline-based scaffold with antiplatelet activity were designed and synthesized on the basis of previous quinoxaline analogues, and the structures were confirmed by ¹H NMR, ¹³C NMR, and MS. The antiplatelet activity was evaluated, structure-activity relationship (SAR) study was summarized and the selectivity of PAR4 was confirmed by calcium mobilization assays. It was indicated that compound 14a, 14g, 13i, 13p showed moderate activity against PAR4, especially, the activity of compound 14g (IC₅₀ = 0.26 μmol·L^-1) was 6.7 times than the lead compound A (IC₅₀ = 1.73 μmol·L^-1). Therefore, 2,3-dihydro-[1,4]dioxino[2,3-g]quinoxaline and [1,3]dioxolo[4,5-g]quinoxaline derivatives are promising compounds for the discovery of novel antiplatelet agents. It is worthy of further research to develop highly effective and selective PAR4 antagonists.

Objective To investigate the apoptosis of T lymphocytes,the expression of Bcl-2, Fas on the peripheral blood T lymphocytes in end stage renal disease patients;and to explore the characteristics of Th1 /Th2 profile and the influence of dialysis membranes with different permeability on the apoptosis of T lymphocytes of maintenance hemodialysis patients.Methods The study included 10 non-dialyszed (ND)patients,45 maintenance hemodialysis patients with cellulose acetate (CA) membranes(13),low-flux polusulfone(PS-LF) membranes(16),high-flux polusulfone (PS-HF) membranes (16) and 8 healthy volunteers (C).The apoptosis of T lymphocytes,expression of Bcl-2,Fas on peripheral blood T lymphocytes cultured with phytohemagglutinin (PHA) stimulation for 24 hours were measured by flow cytometry and immunohistochemical.ELISA was performed for detecting the levels of IFN-?and IL-4 in culture supematants.Results In ESRD patients,the apoptosis of T lymphocytes was greater than that of group C.Group CA was greater than group PS-HF and group PS-LF (P＜0.05).The expression of Bcl-2 on T lymphocytes in ESRD patients was lower than that of group C (P＜0.05).There was negative correlation between the T lymphocytes apoptosis and Bcl-2. The expression of Fas on T lymphocytes in ESRD patients was greater than that of group C (P＜0.05), and it was positive correlated with T lymphocytes apoptosis.The level of IFN-?of ESRD patients was decreased significantly compared with that in group C (P＜0.05),and there was negative correlation between T lymphocytes apoptosis and IFN-?.IL-4 was increased in ESRD patients (P＜0.05) and it was positive correlated with T lymphocytes apoptosis.Conclusions The accelerated apoptosis of T lymphocytes in ESRD patients may be related to the expression of Bcl-2 and Fas of T lymphocytes.ESRD patients show a suppressed secretion of IFN-?and an increased secretion of IL-4. T lymphocytes apoptosis of maintenance hemodialysis patients is influenced not only by the biocompatibility but also by the permeability of the dialysis membrane.

This study aims to investigate the genetic characteristics of BZLF1 gene and its promoter Zp of the epidemic strains in children with primary Epstein-Barr virus (EBV)-associated diseases. Total DNA was extracted from the peripheral blood of 134 children with EBV-associated infectious mononucleosis (EBV-IM) and 32 children with EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH) who were admitted to Beijing Children's Hospital from 2006 to 2011. The EBNA3C, BZLF1, and Zp genes were amplified by PCR assay. Typing of EBV was performed according to the size of the amplification product of EBNA3C gene; the amplification products of BZLF1 and Zp genes were subjected to direct sequencing, and sequence analysis was performed using BioEdit 7. 0. 9. The results were as follows: (1) EBV-1 was present in 140 samples (97.2%, 140/144) and EBV-II in 4 samples (2.8%, 4/144). (2) Three BZLF1 genotypes and their 12 subtypes (including 6 newly found subtypes) were detected in this study; there were no significant differences in the frequencies of BZLF1-A and BZLF1-B between the children with EBV-IM and EBV-HLH (P = 0.083); BZLF1-A1 was the dominant genotype in children with EBV-associated diseases; t BZLF1-A mostly had three 29-bp repeats in the first intron of BZLF1 gene, and BZLF1-B mostly had 30-bp repeats (P = 0.000), with the number of repeats varying from 1 to 13. (3) Four Zp genotypes were detected in this study, including Zp-P, Zp-V3, Zp-V4, and Zp-V1; there were no significant differences in the frequencies of these Zp genotypes between children with EBV-IM and EBV-HLH (P = 0.272, 0.252, 1.0, and 1.0, respectively). (4) The linkage analysis of BZLF1 gene and its promoter Zp showed that BZLF1-A1 was highly associated with Zp-V3 (P = 0.000), while BZLF1-B4 with Zp-P (P = 0.000); EBV-I + BZLF1 A1 was highly associated with Zp-V3 (P = 0.000), while EBV-I+BZLF1-B4 with Zp-P (P = 0.000). The conclusions are as follows: (1) BZLF1-A1 is the dominant genotype in children with EBV-associated diseases; there are mostly 29-bp repeats in the first intron of BZLF1 gene for BZLF1-A genotype and 30-bp repeats for BZLF1-B genotype. (2) Zp-P and Zp-V3 are dominant Zp genotypes of EBV in children, which shared similar detection rates. (3) BZLF1-A1 is highly associated with Zp-V3, while BZLF1-B4 with Zp-P; EBV-I+BZLF1-A1 is highly associated with Zp-V3, while EBV-I+BZLF1-B4 with Zp-P.
Child ; Child, Preschool ; China ; epidemiology ; Epstein-Barr Virus Infections ; epidemiology ; virology ; Female ; Genotype ; Herpesvirus 4, Human ; genetics ; physiology ; Humans ; Infant ; Infant, Newborn ; Introns ; genetics ; Male ; Promoter Regions, Genetic ; genetics ; Repetitive Sequences, Nucleic Acid ; genetics ; Trans-Activators ; genetics

BACKGROUNDIschemic disease is one of the leading causes of death in the world. In order to further study gene therapy for ischemic disease, we constructed a recombinant plasmid for co-expression of human angiopoietin-1 and vascular endothelial growth factor 165(VEGF165) gene in adeno-associated virus (AAV) gene delivery system.

METHODSHuman angiopoietin 1 and VEGF165 gene were obtained using PCR. The upstream of angiopoietin 1 contained restriction enzyme site HindIII, and the downstream of angiopoietin 1 contained restriction enzyme site BamHI. The upstream of VEGF165 contained restriction enzyme site BglII, and the downstream of VEGF165 contained restriction enzyme site BamHI. Using the multiple cloning sites (MCS) in plasmid pZero++ such as BamHI, BglII, HindIII, NotI, XhoI, XbaI, SalI, BspHI, KspI and the corresponding MCS in plasmid pAAV-MCS, angiopoietin 1 and VEGF165 gene were subcloned into pAAV-MCS.

RESULTSDNA sequencing revealed that the PCR- amplified angiopoietin 1 and VEGF165 were consistent with NCBI Gene Bank. The recombinant plasmid was identified using PCR and digestion, which proved to be consistent with our hypothesis. In recombinant plasmid, angiopoietin1 and VEGF possessed a CMV promoter and polyA terminator system respectively, thus assuring co-expression of the two genes.

CONCLUSIONSuccessful construction of AAV co-expression system for human angiopoietin 1 and VEGF165 gene will provide the foundation for gene therapy to cure severe ischemic disease.

Angiopoietin-1 ; genetics ; Dependovirus ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Plasmids ; Vascular Endothelial Growth Factor A ; genetics

OBJECTIVETuberculosis remains a severe public health issue, and the Beijing family of mycobacterium tuberculosis (M. tuberculosis) is widespread in East Asia, especially in some areas in China, like Beijing and Tianjin. This study aimed at determining the mutation patterns of drug-resistant Beijing strains of M. tuberculosis isolated from Tianjin, China.

METHODSA total of 822 M. tuberculosis isolates were screened for drug resistance by an absolute concentration method and the genotype was identified by PCR. 169 drug-resistant isolates of the Beijing family were analyzed for the potential mutations in the rpoB, katG, inhA promoter region and in rpsL, rrs and embB genes, which are associated with resistance to rifampin (RFP), isoniazid (INH), streptomycin (SM) and ethambutol (EMB) respectively by PCR and DNA sequencing.

RESULTSFifty-eight out of 63 RFP-resistant isolates were found to carry the mutations within the 81-bp RFP resistance determining region (RRDR) of the rpoB gene and the most frequent mutations occurred at codon 531 (44.4%), 526 (28.6%), and 516 (7.9%) respectively. 16 mutation patterns affecting 12 different codons around the RRDR of rpoB were found. Of 116 INH-resistant isolates, 56 (48.3%) had the mutation of katG 315 (AGC-->ACC) (Ser-->Thr), 3 (2.6%) carried S315N (AGC-->AAC) and 27 (16.0%) had the mutation of inhA-15A-->T. 84 out of 122 SM-resistant isolates (68.9%) displayed mutations at the codons 43 or 88 with AAG-->AGG (Lys-->Arg) of the rpsL gene and 22 (18.0%) with the mutations at positions 513A-->C, 516C-->T or 905 A-->G in the rrs gene. Of 34 EMB-resistant isolates, 6 had mutation with M306V (ATG-->GTG), 3 with M306I (ATG-->ATT), 1 with M306I (ATG-->ATA), 1 with D328Y (GAT-->TAT), 1 with V348L (GTC-->CTC), and 1 with G406S (GGC-->AGC) in the embB gene.

CONCLUSIONThese novel findings extended our understanding of resistance-related mutations in the Beijing strains of M. tuberculosis and may provide a scientific basis for development of new strategies for diagnosis and control of tuberculosis in China and other countries where Beijing strains are prevalent.

Base Sequence ; China ; DNA Primers ; Drug Resistance, Microbial ; Mycobacterium tuberculosis ; genetics ; Polymerase Chain Reaction