Objective To observe the expression of HBV-DNA and whether there were copy of HBV in the umbilical cord tissues of the died fetus delivered from puerpera with hepatitis virus B. Methods 40 cases such died fetus were collected by routine autopsy to obtain umbilical cord tissues.And using in situ molecular hybridization technique detected HBV-DNA. Results For the umbilical cord tissues, there were 40%(16/40) cases detected out HBV-DNA.HBV-DNA mainly localization in the surface of the umbilical cord vessel and in the cytoplasma of the cord vessel's endothelial cells. They were not in the cord vessel's endothelial cells nuclei. Conclusions There were HBV replication in the umbilical cord tissues of the died fetus. But the expression of HBV-DNA in the umbilical cord tissues of the died fetus is not related with the HBV replication status in the pregnant woman veins.

0.05),but in ages(P

Objective To investigate the clinical features,magnetic resonance imaging (MRI),treatment,and follow-up of patients with glutaric aciduria type Ⅰ (GA-1).Methods Four pediatric patients with GA-1 diagnosed in our hospital were included in this study.They were treated with special diets and carnitine supplements.MRI and tandem mass spectrometry (MS/MS) were performed,and the mental development indices were measured.Results GA-1 was confirmed 2 months,13 months,4 months,and 7 months after birth.Seizure had been observed before the disease diagnosis in three patients and disappeared after treatment.In all four patients,T2-weighted brain MRI showed frontotemporal atrophy or hypoplasia and enlarged subarachnoid space in the sylvian fissures and anterior to the temporal lobes.Diffusion weighted imaging revealed high-density lesions over both the putamen and globus pallidus.The patients were followed up for 4 to 5 years.Plasma amino acids and acylcamitine profile were monitored every 3-5 months.The mean C5DC level and C5DC/C8 were kept the higher limits of the normal ranges,especially in case 3.During the follow-up,the body weight was at-2 SD-0 and the height at-1 SD-0.Intellectual development test showed that case 1 and case 4 had mildly abnormal intelligence,whereas case 2 and case 3 had extremely severe intellectual disability.Gene test confirmed the presence of gene mutations in all four cases,including IVS10-2A ＞ C homozygous mutation in cases 1,3,and 4 and [IVS10-2A ＞ C] + [c.245G ＞c(p.Arg82Pro)] hybrid mutation in case 2.Two female children were smoothly enrolled by local kindergarten,while two male children were unable to walk alone due to delayed motor development and spastic paralysis.Conclusions The phenotype of GA-1 patients is not remarkably correlated with its genotype correlation.Newborn screening is essential for identifying GA-1 patients.

Objective To investigate the role of transforming growth factor-β1 (TGF-β1), Smad2/3 and Smad7 expressions in carotid artery remodeling in renovascular hypertensive rats, and also the therapeutic effect of Enalapril and Amlodipine. Methods The renovascular hypertensive rat (RHR) models with two-kidney and one-clip were established, including model group (n = 6), sham-operated group (n = 6), Enalapril group (10 mg/kg per day, n = 6), Amlodipine group (5 mg/kg per day, n = 6) and combination group (Amlodipine 2.5 mg/kg per day + Enalapril 5mg/kg per day, n = 6). The medication were continuous administrated for six weeks. Carotid artery morphological and structural changes in the media were observed by HE staining, Masson staining and immuno histochemical staining. Media thickness (MT), MT and lumen diameter ratio (MT/LD), and the expression levels of media α-smooth muscle actin (α-actin), proliferating cell nuclear antigen (PCNA), TGF-β1, phosphorylated Smad2/3 (p-Smad2/3) and Smad7 in carotid arteries were measured. Results The media of carotid arteries in RHR model group was significantly thickened, the volume of smooth muscle cell was increased, and the array was in disorder; MT, MT/LD, the proliferation index of smooth muscle cell and collagen fiber area percentage of carotid arteries in the model group were significantly higher than those in the sham-operated group (P < 0.01). Compared to sham-operated group, the model group had significantly higher expressions of TGF-β1 and p-Smad2/3 (P < 0.05) and lower Smad7 expression. Both Enalapril and Amlodipine improved smooth muscle hypertrophy and collagen deposition, reduced RHR carotid MT, MT/LD, proliferation index of smooth muscle cell, collagen fiber area percentage and the expressions of TGF-β1 and p-Smad2/3 (P < 0.05), increased Smad7 expression (P < 0.05). Moreover, the combination treatment of Enalapril and Amlodipine had significantly better effects than single Amlodipine group (P < 0.05), but not single Enalapril group. Conclusions TGF-β1/Smads pathway may participate in the mechanism of carotid artery remodeling in RHR; the role of Amlodipine and Enalapril in inversing carotid artery remodeling may be related to the change of TGF-β1/Smads pathway, the combination treatment of Amlodipine and Enalapril had better effects than single administration of Amlodipine.

BACKGROUND:Poly(D,L-lactide-co-glycolide)(PLGA)may produce lactic acid and glycolic acid when PLGA degrades,thus leading to the acid accumulation and inducing the inflammatory reaction locally.OBJECTIVE:To investigate the effect of lysine,histidine and arginine on regulating the acid accumulation of PLGA copolymer during the degradation.DESIGN:Repeated measurement and experiment.TIME AND SETTING:The experiments were performed in the College of Bioengineering.Chongqing University from July 2006 to August 2007.MATERIALS:PILGA(80:20)was produced by Sigma(USA);Lysine,histidine and arginine(purlty>99%)were purchased from Sigma(USA);Chitosan(deacetylation degree:85%)was purchased from Chengdu Kelong Chemical Reagent Factory;Algin(viscosity:1.05-1.15)was purchased from Tianiin Damao Chemical Reagent Factory.METHODS:Lysine,histidine and arginine were added into PLGA with the proportion of 5%(w/v)and 10%(w/v)respectively.The resultant film sample was put into a bottle with tri-distilled-water for 2-month degradation at 37℃.The pH value of degradation solution was detected by pH meter;Each film sample was taken out and lyophilized 12 hours to get its dry weight and calculate mass loss ratio.Each variety of the solution was sampled three specimens,the average pH value,average initial weight and average finial weight of these three specimens were taken as the indices at each sampling time point,respectively.Accordingly,the regulation effect of basic amino acid was comparexl with that of algin,chitosan and NaHCO3.MAIN OUTCOME MEASURES:The changes of pH value of degradation solution;the mass loss ratio of the composite.RESULTS:Each basic additive could relieve the acid accumulation,among them,NaHCO3 was the strongest,while algin and chitosan showed a lowest capacity,basic amino acid was moderate;The suitable regulation effect could be achieved at a level of 5%lysine.CONCLUSION:Basic amino acid can effectively regulate the acid accumulation after PLGA degrades,and the optimal concentration of lysine is 5%.

Adult ; Female ; Hepatitis B Core Antigens ; metabolism ; Hepatitis B Surface Antigens ; metabolism ; Hepatitis B, Chronic ; drug therapy ; immunology ; virology ; Humans ; Liver ; metabolism ; Male ; Middle Aged ; Vidarabine ; therapeutic use

Objective To compare the immune effects of Coxsackievirus B3 (CVB3) capsid protein VP1 expressed bacterially, recombinant adenovirus rAd/VP1 and recombinant plasmid pcDNA3/VP1which express VP1 protein in mice. Methods After expressed in prokaryotic cells, VP1 protein was purified. Recombinant adenovirus rAd/VP1 and recombinant plasmid pcDNA3/VP1 were amplified and extracted. Six to 8-week-old, male BALB/c mice were divided into four groups randomly. Each group contained 18 mice. The mice of pcDNA3/VP1 group or VP1 protein group were immunized intramuscularly with three injections at three weeks apart, of recombinant plasmid pcDNA3/VP1 at a dose of 100 μg/mouse or recombinant protein VP1 at a dose of 50 μg/mouse. The mice of rAd/VP1 group were immunized intramuscularly twice at two weeks interval with rAd/VP1 at a dose of 1.2 × 107 PFU. The control group was mock-immunized with 100 μl of PBS intramuscularly. Mice were bled from the retroorbital sinus plexus every two weeks after each immunization. ELISA and micro-neutralization test were used to detect levels of CVB3-specific IgG antibody and neutralizing antibody titers in the sera of immunized mice. Three weeks after the last immunization, the cytotoxic T lymphocyte(CTL) killing activity of spleen lymphocytes was detected with CCK-8 assay. Subsequently, virus titers in the sera of immunized mice were determined by the 50% cell culture infective dose( CCID50 ) assay on HeLa cell monolayers and percentage of animals surviving were observed after lethal CVB3 attack over a period of 21 days. Results The titers of specific IgG antibody and neutralizing antibody in sera of VP1 protein immunized mice were higher than other groups( P ＜0.05 ). While CTL killing activity of spleen lymphocytes of VP1 protein immunized mice was lower than mice in rAd/VP1 group( P ＜0. 05). Virus titers in sera of VP1 protein immunized mice were lower than the mice in pcDNA3/VP1 or rAd/VP1 groups ( P ＜ 0.05 ), while survival rate was significantly higher than these two groups ( P ＜ 0.05 ).Conclusion VP1 protein induced higher level of humoral immune response and acquired obvious immune protection effects in mice. The immunizing potency of VP1 protein vaccine surpassed plasmid pcDNA3/VP1or recombinant adenovirus rAd/VP1. It appeared to be a promising candidate among the three different vaccines.

OBJECTIVETo explore the differential gene expressions of chronic hepatitis B (CHB) of Gan depression Pi deficiency syndrome (GDPDS) and Pi-Wei damp-heat syndrome (PWDHS).

METHODSThe ulnar venous blood was withdrawn from healthy subjects (26 cases), patients of GDPDS (35 cases) and PWDHS (34 cases) on an empty stomach. The total RNA was extracted using Trizol method. The differential genes were detected using Aglient expression profile chip and screened using randomized variance model. The results were analyzed using GO, Pathway, GeneBank, NCBI, and Geneontology.

RESULTSThere were 125 differential genes between CHB patients of GDPDS and those of PWDHS (including 66 up-regulated genes and 59 down-regulated genes), mainly involving in functions of transmembrane transport, response to selenium ion, and regulation of calcium ion-dependent exocytosis. The signal pathway participated in mainly includes cell adhesion molecules, calcium ion signaling pathway, leukocyte trans-endothelial migration. We present gene co-expression networks to find 9 interactions among genes (LOC340508, HIST2H2BE, MPL, FLJ22536, TUBA8, NT5M, EGFL7, PTPRF, TSPAN33), which were mainly involved in immune response, cell growth, DNA damage, signal transduction, inflammatory reaction, and so on.

CONCLUSIONSThe differential expression genes existed between CHB patients of GDPDS and those of PWDHS, indicating that Chinese medicine syndrome classification has its own basis for gene expression profile. The genomics research method is expected to provide an objective basis for Chinese medicine syndrome typing.

Adult ; Case-Control Studies ; Female ; Gene Expression ; Hepatitis B, Chronic ; diagnosis ; genetics ; Humans ; Male ; Medicine, Chinese Traditional ; methods ; Oligonucleotide Array Sequence Analysis ; Transcriptome

OBJECTIVE: To determine the type and carrier rate of deafness-related variants in Dongguan, China. METHODS: A total of 16 182 subjects were screened. Heel blood samples were collected from newborns, while peripheral venous blood samples were collected from the remainders. For each individual, 100 variations of 18 deafness susceptibility genes were detected. RESULTS: In total 1631 deafness-related variants (including 5 homozygous mutations) were detected, which gave a detection rate of 10.08%. The detection rate of SLC26A4 gene variants was the highest (845 cases, 5.22%), which was followed by GJB2 (673 cases, 4.16%), GJB3 (100 cases, 0.62%), TMC1 (12 cases, 0.07%), and MYO15A (1 case, 0.01%). The detection rate for GJB2 c.235delC variant was the highest (524 cases, 3.24%), which was followed by SLC26A4 IVS7-2A>G variant (270 cases, 1.67%). Thirty three individuals (0.20%) carried two variants at the same time, 7 of them (0.04%) carried compound heterozygous variants of the same gene. CONCLUSION To expand the range of screening can help with determination of the carrier status and provision of early intervention and genetic counseling for the examinees.
China ; DNA Mutational Analysis ; Deafness ; genetics ; Genes ; Genetic Counseling ; Genetic Predisposition to Disease ; Genetic Testing ; Genetic Variation ; Humans ; Infant, Newborn ; Mutation ; RNA, Ribosomal

Objective To analyze the risk factors of hospital acquired pneumonia (HAP) caused by nonfermentative bacteria in ICU.Methods Using binary logistic regression analysis,we reviewed the related risk factors of hospital acquired pneumonia caused by nonfermentative bacteria from January,2002 to May,2006 in ICU of our hospital.Results Univariate regression ana- lysis identified 22 variables associated with HAP,including age,diabetes mellitus,malignant tumor,chronic obstructive pul- monary disease (COPD),smoking,Glasgow Coma Scale (GCS) score,Acute Physiology and Chronic Healthy EvaluationⅡ(APACHEⅡ) score.Multivariate regression analysis confirmed 6 high risk factors:aspiration,COPD,use of antacids,seda- tive agents,mechanical ventilation and tracheal intubation or tracheostomy.Conclusions Aspiration,COPD,use of antacids, sedative agents,mechanical ventilation,tracheal intubation or tracheostomy are high risk factors associated with HAP.