To investigate the potential role of hepatitis B virus core protein (HBcAg) in the pathogenesis of hepatitis B and seek for an effective therapeatic approach to the treatment of hepatitis B, the improved yeast two hybrid technique was employed to construct HBcAg bait plasmid, and the plasmids were transformed into yeast AH109. Then the yeast AH109 was mated with yeast Y187 containing liver cDNA library plasmid in 2?YPDA medium, diploid yeast was plated on both synthetic dropout nutrient medium (SD/ Trp Leu His Ade) and synthetic dropout nutrient medium (SD/ Trp Leu His Ade) containing X ? gal to be selected and screened two times. Plasmids were extracted from sixteen positive colonies, and they were sequenced. Among them, two colonies contained metallothionein. The interaction between HBcAg and metallothionein was confirmed by in vitro reticulocyte lysate translocation and immunoprecipitation. The results suggested that the pathogenesis of hepatitis B metallothionein might play an important role in the episode process.

To investigate the biological functions of hepatitis B virus e antigen (HBeAg) on pathogenesis of HBV, and to seek for effective methods to prevent and cure hepatitis B, we performed the yeast two hybrid system 3 technique to construct HBeAg bait plasmid. The bait plasmid was transformed into yeast AH109 and expressed in it. After the expression of HBeAg was identified by SDS page and Western blot, the AH109 yeast was mated with yeast Y187 containing liver cDNA library plasmid in 2 YPDA medium to form diploid yeast and plated on synthetic dropout nutrient medium (SD/ Trp Leu His Ade) and synthetic dropout nutrient medium (SD/ Trp Leu His Ade) containing x ? gal for screening. Plasmids of blue colonies were extracted and transformed into Escherichia coli, then analyzed by DNA sequencing and bioinformatics. Thirty nine positive colonies were sequenced, among them three colonies were metallothionein. To further prove the interaction between HBeAg and metallothionein, translation was performed by using reticulocyte lysate, and immunoprecipitation was showed in vitro . The results indicated that hepatitis B virus e antigen could interact with metallothionein in vivo and in vitro.

Abruptio Placentae ; complications ; Adult ; Disseminated Intravascular Coagulation ; diagnosis ; etiology ; therapy ; Female ; Humans ; Infant, Newborn ; Male ; Pregnancy ; Prognosis ; Risk Factors

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Constipation ; physiopathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Young Adult

Aged ; Humans ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Pneumoconiosis ; drug therapy ; microbiology ; Tuberculosis, Multidrug-Resistant ; drug therapy ; microbiology

Objective To develop a system failure recovery programs in order to prevent an accident of the hospital computer network system. Methods The network failure was divided into three grades according to its influence. The groups of network malfunction was set up and can immediately start the established program, take effective measures to prevent the situation expand and spread in the event of unforeseen circumstances. Results Through the emergency power supply program, the network route emergency program, cable emergency support program and sector emergency plan, the hospital information systems run normally. Conclusion The issue of large-area network information paralysis is effectively solved and improves the information technology capacity of the hospital.

The structure, reaction mechanism and molecular biology of Leuconostoc mesnteroides dextransucrase are reviewed.

Objective To identify the arteriosclerosis (AS)changes in peripheral artery and abdominal aorta of patients with familial hypercholesterolemia(FH) during follow-up.Methods Seventeen patients of 6 FH [5 male and 12 female with average age of (16.12?6.65) years old],along with 17 subjects of matching sexes and ages with normal blood cholesterol as healthy control group,underwent examination by color doppler ultrasound,and changes of intima-media thickness (IMT) in peripheral artery and abdominal aorta,severity of stenosis,morphology,and function were observed.Results For 17 patients of FH,the total cholesterol(TC),low density lipoprotein cholesterol(LDL-C) in serum were higher significantly and high density lipoprotein cholesterol (HDL-C) was lower significantly than those in healthy control group(Pa0.05).The IMT of carotid aorta,subclavicular aorta,common abdominal artery,and common femoral artery in 13 patients were showing various degrees of increase,yielding an average of 2.9 mm.As patients aged,this phenomenon was reported to be more profound in common abdominal aorta and femoral artery.Conclusions Patients of FH show AS lesion in early stage and it worsened as they aged,from carotid arteries to common abdominal aorta and femoral artery.Color doppler ultrasound can be a non-invasive examination for monitoring the progress of AS in blood vessels in patients of diagnosed FH.

Objective To identify mutations site and clinical characteristics of a familial hypercholesterolemia(FH) proband diagnosed clinically through DNA sequencing and family analysis in the proband and his family members of 3 generations.Methods Blood samples and clinical data of the kindred of total 29 from 3 generations members were collected.Proband had a physical examination electrocar-diogrom and vascular ultrasound.The proband and his family members took routine clinical exams,and genomic DNA was isolated.The promoter region and the 18 exons of low density liporotein receptor(LDLR) gene were screened by Touch down polymerase chain reaction -single strand conformation polymorphism(PCR-SSCP) and DNA sequencing.The result of sequencing were matched gene sequence published in the BLAST database.Results 1.Increased intima-media thickness and plaque were detected in the common carotid artery,right subclavian artery of the proband.Aortic valve regurgitation was found by echocardiography.2.No mutation R3500Q of ApoB100 was observed.3.Two heterozygous mutations in exon 10 and 13 of LDLR gene (W462X and A606T) were identified.The proband and 5 members of paternal relatives showed W462X heterozygosis mutation in exon 10 of LDLR gene which introduced the change from tryptophone to a new stop codon.The proband's mother and grandmother harboured A606T heterozygous mutation in exon 13 of LDLR gene due to a single base pair substitution of G for A in the codon for residue 1 879.Conclusions Disease causing mutations of proband are W462X and A606T compound heterozygosis mutation in exon 10 and 13 of LDLR gene inherited from mother and father.Proband shows homozyous phenotype though the genotype analysis indicates heterozygous mutations.

Animals ; Disease Models, Animal ; Hypoglycemia ; chemically induced ; metabolism ; Hypothalamus ; drug effects ; metabolism ; Insulin ; pharmacology ; Intracellular Signaling Peptides and Proteins ; metabolism ; Male ; Neuropeptides ; metabolism ; Orexins ; Rats ; Rats, Sprague-Dawley