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.

Background Noninvasive methods such as in vivo confocal microscopy and Orbscan Ⅱ corneal topography have been used to examine ocular surface structure at the cellular level.However,very few domestic reports about the corneal structures of experimental animals investigated by confocal microscopy are available.Objective This study was to compare the anatomical differences of the corneal structures of three frequently used experimental animals presented by in vivo confocal microscopy,and to offer a database on the information provided by the in vivo study of the corneal structures of these animals.Methods Bilateral corneas of 3 clean adult male New Zealand rabbits,3 clean adult male Lewis rats and 3 clean adult male Swiss mice were examined by in vivo confocal microscopy.The morphological characteristics of every layer of the corneas and the endothelial cell densities were analyzed and compared.Results Superficial epithelium cells of the three animal models were characterized as polygon cells with high or low reflective border.The arrangement of the basal epithelial cells was regular with tight contacts but these cells lacked visible nuclei.The Bowman' s layer of cornea presented as an amorphous sheet containing abundant subepithelial plexus.In the rabbits,a highly reflective structure in the corneal stroma wasconfirmed as the nucleus,and the cell density of the posterior stroma was significantly lower than that of anterior stroma(387.5 cells/mm2 versus 223.5 cells/mm2)(U =0.000,P =0.000).Massive light-reflecting astreoids were displayed in the stroma of the rats and the mice.Corneal endothelial cells(CECs)of the three animal models had similar shapes and arrangements,presenting with high refractive cell bodies with dark borders and honeycomb-like arrangements.The CECs densities were 2192.5,1936.0,1565.0 cells/mm2 in the New Zealand rabbits,Lewis rats and Swiss mice,respectively,showing a statistically significant difference among them(H =49.940,P =0.000),and that of the rabbits was significantly higher than that in the rats and mice(x2 =0.000,P =0.000;x2 =0.000,P=0.000).Significant difference was also seen between the rats and the mice in the CECs densities(x2=0.000,P=0.000).Conclusions The CECs of the three animal modes are similar in morphology.But the structures of their stromal cells and endothelial cell densities are different.The combination of in vivo confocal microscopy and Orbscan Ⅱ corneal topography offers high-resolution imaging for each layer of the cornea.

AIM: To observe the clinical effect of the tonifying kidney pills with mingmuwuzi treating evaporative dry eyes.METHODS: This study adopted the positive drug control,prospective study,random number remainder grouping method to 65 cases of outpatient patients diagnosed with evaporative dry eyes which were divided into the treatment group 32 cases (64 eyes) and the control group 33 cases (66 eyes).The treatment group took the decoction of kidney pills with mingmuwuzi,combined with sodium hyaluronate eye drops.The control group simply use sodium hyaluronate eye drops,both group were set to 4wk for a course of treatment.To observe the symptoms and signs of two groups before and after the treatment,the change of the evaluation index and curative effect were evaluated.RESULTS: The effectiveness of the treatment group was 87.5%,the control group was 78.8%,the difference was statistically significant (z=-3.149,P<0.05).CONCLUSION: The treatment of the kidney pills with mingmuwuzi combined with sodium hyaluronate eye drops to evaporative dry eyes is more effective than the simple use of sodium hyaluronate eye drops.

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.

ObjectiveTo observe the expression of Caspase-3 mRNA in prefrontal cortex and hippocampus of chronic stress-induced depression rats,and to detect the machnisms of antidepression by electro-acupuncture.MethodsSprague-Dawley rats were randomly divided into four groups:control group,model group,model +electro-acupuncture group and model + paroxetine group,12 rats in each group.Open-field test was used to observe the changes of movements,and real-time fluorescence quantitative PCR (RT-PCR) method was used to detect Caspase-3 mRNA levels in prefrontal cortex and hippocampus.Results ①Open-field test:after stress,compared with control group rats,the model group rats' crossing numbers (29 ± 7),rearing times (6 ± 2) were apparently less than those of control group( (66 ± 13),( 10 ±2) ; P＜0.05,P＞0.05).In comparison with model group,the crossing times and rearing times being increasing in degree in electro-acupuncture group( (61 ±9),( 13 ±1 ) ) and paroxetine group( (39 ± 10),(8 ± 1 ),P＜0.01,P＞0.05).② Compared with the control group,Caspase-3 mRNA in prefrontal cortex and hippocampus significantly increased in model group(P ＜0.05 ) ;and compared with model group,Caspase-3 mRNA in prefrontal cortex in electro-acupuncture group and paroxetine group significantly decreased(P ＜ 0.05 ),and both expression of Caspase-3 mRNA in hippocampus in electro-acupuncture group and expression of Caspase-3 mRNA in hippocampus in paroxetine group decreased (P ＞ 0.05 ).ConclusionChronic stress can increase the expression of Caspase-3 mRNA in prefrontal cortex and hippocampus of chronic stress-induced depression rats,while electro-acupuncture can decrease the expression of caspase-3 mRNA,which may be an important way to anti-depression by electro-acupuncture.

Biomarkers ; metabolism ; Cholesterol ; metabolism ; Coronary Disease ; Humans

Astrocytoma ; genetics ; metabolism ; pathology ; Brain Neoplasms ; classification ; genetics ; metabolism ; pathology ; Child ; Child, Preschool ; Chromosomal Proteins, Non-Histone ; genetics ; metabolism ; Chromosome Deletion ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; DNA-Binding Proteins ; genetics ; metabolism ; Ependymoma ; genetics ; metabolism ; pathology ; Hedgehog Proteins ; genetics ; metabolism ; Humans ; Medulloblastoma ; classification ; genetics ; metabolism ; pathology ; Proto-Oncogene Proteins B-raf ; genetics ; metabolism ; Rhabdoid Tumor ; genetics ; metabolism ; pathology ; SMARCB1 Protein ; Signal Transduction ; Transcription Factors ; genetics ; metabolism ; Wnt Proteins ; metabolism ; beta Catenin ; genetics ; metabolism

The study evaluates the lipolysis rate and extent of type Ⅲ lipid formulations using testosterone undecanoate as a model drug after digestion with in vitro lipolysis model, and studies the digestive regularity with optical microscope and electrical conductivity. The results showed that for testosterone undecanoate type Ⅲ lipid formulations with castor oil as oil phase and Transcutol HP as latent solvent, the lipolysis rate and extent were increased with the increase of oil phase proportion and were decreased with excessive proportion of surfactant, in which can see liquid crystal phase during lipolysis process. The lipolysis rate of type ⅢB lipid preparations with different surfactant were ordered as Labrasol > Tween 80 > Cremophor EL, but the rate of type ⅢA is different in quick digestion phase and slow digestion phase. The lipolysis extent of type Ⅲ lipid formulations with different surfactant were ordered as Cremophor EL > Tween 80 > Labrasol. These may be related to the digestive effect of pancreatic lipase on different surfactants. This study implied that the lipolysis rate and extent of type Ⅲ lipid formulations are greatly influenced by the proportion of oil phase and surfactant, and the surfactant structure. These factors will affect the in vivo digestion and should be taken into account when screening type Ⅲ lipid formulations.

Animals ; Arsenic ; toxicity ; Comet Assay ; DNA Damage ; drug effects ; Lymphocytes ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Tobacco Smoke Pollution ; adverse effects