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BACKGROUND: Recently, one focus of research has been Annexin Ⅴ (AnV) existing on hepatic cells membranes as a fundamental receptor related to hepatitis B virus (HBV) infection. Also its expression in placental tissues has been a matter of debate. The study of the relationships between placental cells infected with HBV and their AnV expression will be of great value in future prevention strategies and treatments.OBJECTIVE: To investigate the presence of AnV in HBV infected human's placental cells and its potential role in HBV intrauterine transmission.DESIGN: Randomized controlled experiment.SETTING: Taishan Medical College.MATERIALS: Placental tissue was collected from HBsAg positive full term pregnant women (30 cases) admitted to Jinan Institute for Maternal and Child Health, Taian Central Hospital and Taian Institute for Maternal and Child Health from January 2003 to December 2004. Maternal serum was also obtained. Informed consents for participating in this study were obtained from all the involved pregnant women and this experiment was approved by the Hospital Ethics Committee. Rabbit-anti-human AnV purified affinity antibody (first antibody), rat-anti-human HBs mAb (first antibody),and biotinylated goat-anti-mouse IgG (secondary antibody) were supplied by Wuhan Boster Bioengineering Company.METHODS: Using SABC immunohistochemical staining reagent, 18 HBsAg positive placentas were obtained from 30HBsAg infected patients in full term pregnancy. These were considered as the positive group and the other 12 were used as negative controls. The staining process included dewaxing, dehydration of embedded slides and microwave antigen restoration. In the wet box, rabbit-anti-human AnV purified antibody (first antibody, 1:60, monoclonal antibody)was added on the slides and kept at 4 ℃ overnight. Rat-anti-human antibody HBs mAb(secondary antibody, 1:50) was added and kept at 4 ℃ ovemight, after this procedure, biotinylated goat-anti-mouse IgG(1:100), the first fluorescent antibody such as FITC-goat anti-rabbit IgG (1:50) and the second fluorescent antibody (Avidin-Cy3) were used,respectively. The slides were sealed with buffered glycerol and examined under a confocal laser scanning microscope.The images on the slides were analyzed with IPP 4.5 image programs.MAIN OUTCOME MEASURES: Detecting the simultaneous existence and distribution of HBsAg/AnV in placental cells with HBV infection.RESULTS: Ten cases from the positive group were simultaneously detected for HBsAg/AnV by double-labeled immunofluorenscence assay and confocal laser scanning microscope. AnV expression was detected in the trophoblastic, interstitial cells and vascular endothelial cells of villi interstitial blood vessels, and the coexistence of HBsAg/AnV was found even in one cell.CONCLUSION: HBsAg combined with the receptor AnV in the same placental cells is a common finding in HBV infected full term pregnant women. This finding is very suggestive of a mechanism where AnV could promote hepatitis B virus to enter the placental cells and cause intrauterine infection.
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LA G1 was identified as a gene that is differentially expressed during the yeast replicative life span and was shown to play a role in determining yeast longevity. The cDNA of rat LASS1, the mammalian homolog of yeast LA G1, was cloned from rat cerebral cortex and sequenced, which is different to the predicted sequence in the GenBank. Sequence analysis revealed that this cDNA clone contains an open reading frame of 1 053 bp. The deduced amino acid sequence has 350 residues and shares a predicted Laglp motif and a TLC domain conserved in Lag1 proteins. Total RNAs were isolated from rat cerebral cortices at varying ages: newborn, one month, six months, twelve months, and twenty-four months. Semi-quantitative RT-PCR and Northern blot analysis were performed to analyze the LASS1 expression level in rat cerebral cortex tissues at varying ages. Senescence-associated β-galactosidase (SA-β-gal) activity was firstly used as a biomarker for assessing senescence in rat neurons. The results showed that LASS1 expression was upregulated from newborn to adult rats (1～6 month) and declined in aged cortex. SA-β-gal staining positive neurons significantly increased in the aged cerebral cortex. The age-related expression alternation of LASS1 in rat cerebral cortex provides an important clue in exploring the role of LASS1 in mammalian neuron aging.
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Objective To study the role of IL 12 and IL 13 mRNA in children with asthma. Methods Use of semi quantitative RT PCR, IL 12 and IL 13 mRNA in peripheral blood mononuclear cell (PBMC), as well as total IgE in serum from children with asthma, which is in the period of acute phase, were detected. Results Compared with control group, The expression level of IL 12 mRNA were decreased and that of IL 13 mRNA were increased in asthmatic children; The sicker the patient was, the lower expression of IL 12 mRNA, the higher expression of IL 13 mRNA; No matter how the IgE level was, there was significantly different between the expression of IL 12 and IL 13 mRNA. Conclusion IL 12 and IL 13 may be one of the factors causing bronchial chronic inflammation.
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AIM:To study the changes in lymphatic motion and histochemical stain of nitric oxide synthase(NOS) in the early phase of acute rat endotoxemia and explore their relationship. METHODS: After lipopolysaccharide (LPS) was injected into femoral vein, rats' mesenteric lymphatic were inspected within two hours. Paraffin section was stained by HE, frozen section by NADPH diaphorase histochemistry. RESULTS: In LPS group, the mesenteric lymphatic diameter was greater than that in the control and contractile frequency was decreased, the villi central lacteal and submucosal microlymphatics of small intestine was extremely dilated under microscope. In LPS and control group, NOS was positive in lymphatic endothelia and the nerve fibers in the vessels' wall. CONCLUSION: In the early stage of acute endotoxemia, the lymphatic vessles dilated and contractile activity diminished, NOS may play an important role in these changes.
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Objective\ Clone and sequence choriocarcinoma related gene T26. Method\ Choriocarcinoma related gene T26 was ligated into PGEM\|T vector using T\|A clone method.PGEM\|T\|T26 was digested with EcoRⅠ and sequenced,comparing the T26 sequence with GenBank. Results\ The sequence of choriocarcinoma related gene band T26 showed more than 99% identity with the 3' end of human scar,rps4 and ccg2 genes. Conclusion\ The gene T26 besides scar,rps4 and ccg2 genes were related with the oncogenesis of choriocarcinoma. [