Aims To study single dose toxicity of poly-phenols effective parts from Punica granatum,to eval-uate their safety,and thus to provide a theoretical basis for drug development and clinical use.To observe their protective effect on ethanol-induced gastric ulcer in rats.Methods 50 healthy Kunming mice were ran-domly divided into five groups and given different doses of polyphenols’effective parts from Punica granatum via intragastric administration.Toxicity and death in each group of mice were observed and recorded after administration for 14 d.The median lethal dose was calculated by Bliss method.70 rats were randomly di-vided into normal group,model group(constant volume of normal saline),sanjiuweitai particles(1 850 mg· kg-1 )group,colloidal bismuth subcitrate (33 mg · kg-1 )group and polyphenols effective parts from Puni-ca granatum low-dose,medium-dose,high-dose(430, 852,1 704 mg·kg-1 )groups.On the 9th day of 10 days’gavage,all except the normal group were fed ethanol (1.5 mL/only)to induce gastric mucosal inju-ry in rats with acute gastric ulcer.Gastric ulcer index, the rate of ulcer inhibition were calculated for each group.The morphological changes of gastric mucosa were observed.The gastric mucosa levels of PGE2 , NO,SOD and MDA were determined.Results The LD50 and 95%confidence limit of the polyphenols’ef-fective parts from Punica granatum were 8 520.9 mg· kg-1 and 7 291.2 ~9 914.4 mg·kg-1,respectively. Pathology showed that the organs receiving dose of 16 000 mg · kg-1 had different degrees of damage . Compared with the model group,the extract from Puni-ca granatum significantly repaired the gastric mucosa, and significantly increased the gastric mucosa levels of NO and reduced MDA content,and improved SOD content and the levels of PGE2 .Conclousion The dose of 5 063 mg · kg-1 of polyphenols effective part from Punica granatum showed no death.The dose of 16 000 mg · kg-1 of polyphenols effective parts from Punica granatum could cause varying degrees of dam-age in heart,liver,lung,kidney or the death of mice. The LD50 and 95% confidence limit of the polyphenols effective parts from Punica granatum were 8 520.9 mg ·kg-1 and 7 291.2 ~9 914.4 mg·kg-1,respective-ly.The extract from Punica granatum plays a protective role against gastric mucosa damage induced by absolute ethanol,and the mechanism may be related to promo-ting ulcer epithelial cells synthesis,enhancing mucosal regeneration function,regulating NO content and en-hancing antioxidant capacity.

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. [

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.

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.

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.

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.

objective To evaluate the distribution and significance of IgG subclasses of antithyroglobulin antibody in sera from patients with Hashimoto thyroiditis.Methods Sera from 112 patients with Hashimoto thyroiditis were collected and patients were divided into 3 groups,i.e.hypothyroidism,subclinical hypothyroidism and euthyroidism.Antigen specific ELISA was used to detect the distribution of IgG subclasses of antithyroglobulin antibody.Results The positive rates of IgG subclasses of TgAb were IgG1 90.2%,IgG2 58.0%,IgG3 19.6%and IgG4 87.5%respectively.The mean geometric titers of IgG1 in sera from patients with hypothyroidism and subclinical hypothyroidism were 1:450.8 and 1:245.5 respectively,both being significantly higher than that with euthyroidism(1:8.7,P＜0.01).The mean geometric titers of IgG2 in sera from patients with hypothyroidism and subclinical hypothyroidism were 1:37.3 and 1:3.2 respectively,both being also significantly higher than that with euthyroidism(1:0.2,P＜0.01 and P＜0.05 respectively)and that with hypothyroidism was significantly higher than that with subclinical hypothyroidism(P＜0.05).Conclusion The distribution of IgG subclasses of antithyroglobulin antibody in sera from patients with Hashimoto thyroiditis was predominantly IgG1,IgG2 and IgG4.High titers of IgG1 and IgG2 implicated the possibility of development from subclinical hypothyroidism to overt hypothyroidism.

Objective To assess the association of synovial cyclic citrullinated peptide(CCP)expression with T helper 17(Th17) cells/Regulatory T cells (Treg) imbalance,the histological and clinical features of synovitis in rheumatoid arthritis (RA).Methods CCP expression in synovial specimens from 39 patients with RA and 35 controls was detected by immunohistochemistry assay(IH) using 6×His tagged anti-CCP single chain fragment V (ScFv) antibodies,which were generated by pHEN2 phagemid recombinant antibodies display system.The frequencies of Th17/Treg cells in the peripheral blood mononuclear cells (PBMCs) were determined by flow cytometry (FCM).Th17/Treg cells associated cytokines were analyzed by enzyme-linked immunosorbent assay (ELISA).The histological scores and clinical features of synovitis were included in the study.Chi-square test and ANOVA were used for statistical analysis.Results ① The prevalences of synovial C CP expression were significantly different between RA group and the control(76.9％ and 11.4％ respectively,X2=31.9,P＜0.01).② The frequencies of Th17 cells,Th17/Treg ratio,Th17 cells associated cytokines as IL-6,IL-17a,IL-23,TNF-α,and the Treg cells associated cytokines TGF-31,the serum and synovial fluid anti-CCP antibodies in the RA patients with synovial CCP positive expression were significantly higher in RA patients with CCP positive than those with CCP negative.Disease activity score DAS28 index and histological features quantified variations of the synovial biopsy specimens (synoviocyte hyperplasia,focal aggregates of lymphocytes,and diffuse infiltrat(e)s of lymphocytes) in RA were higher in synovial CCP positive expression patients than in the negative.Conclusion Synovial CCP expression is strongly associated with the Th17/Treg imbalance and synovitis,which may play a crucial role in the pathogenesis of RA.

OBJECTIVETo investigate the association between small ubiquitin-related modifier-1 (SUMO-1) gene rs6709162, rs7599810, rs7580433 polymorphism and non-syndromic oral clefting (NSOC).

METHODSOur study consisted of 208 Ningxia NSOC patients, their parents (189 fathers and 176 mothers), 172 nuclear families (patients and their parents), and 284 normal controls. DNA was extracted and polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) was used to identify rs6709162, rs7599810, rs7580433 genotypes of the samples. The data was analyzed by case-control analysis, family based associated test (FBAT), and transmission disequilibrium test (TDT).

RESULTSCase-control study found that TT genotype's frequency was significantly different in cleft lip and cleft palate group compared with the control group at rs7599810 of SUMO-1 (P=0.01, P=0.01). TDT test showed that rs7599810's T allele had over-transmitted (P=0.00) in cleft lip and palate group. FBAT analysis revealed that distribution of rs7599810's TT genotype and T allele was significantly different (P=0.00, P=0.00). TDT test showed that rs6709162's C allele in cleft palate and cleft lip and palate patients had over-transmitted (P=0.00, P=0.01). rs7580433's G allele in cleft lip group had over-transmitted (P=0.05).

CONCLUSIONSUMO-1 gene polymorphism is associated with NSOC.

Case-Control Studies ; Cerebellar Ataxia ; Cleft Lip ; Cleft Palate ; Female ; Genotype ; Humans ; Intellectual Disability ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; SUMO-1 Protein ; genetics ; Ubiquitins

Objective To compare the effects of cleaning oral among different nursing methods for patients with dysphagia after brain injury who take food by gastric tube as well as by mouth. Methods A total of 59 patients with dysphagia after brain injury were divided into research group ( 29 cases, mouth nursing by modified method of brushing their teeth and irrigation ) and control group ( 30 cases, mouth nursing by traditional oral care routine) according to the random number table method from October 2012 to May 2014. Results were observed by comparing oral condition, indexes of pharyngeal swab culture and water-swallowing test. Results Incidence rates of halitosis and unclean mouth were all 13. 8% in the research group which were lower than those (73. 3%, 63. 3%) in the control group with statistically significant differences (χ2 =21. 208, 15. 214;P<0. 01). The effective rate of water-swallowing test was 79. 3% in the research group which was higher than that (56. 7%) in the control group (χ2 =12. 202,P<0. 01). There was no statistically significant difference in abnormity rates between two groups before intervention (P >0. 05). The abnormity rate in the research group was lower than that in the control group at 4th week (χ2 =8. 958,P <0. 01). There was statistically significant difference in abnormity rate before interventions and at 4th week in the research group (P<0.01), with no statistically significant difference in the control group (P>0.05). The rate of oral mucosa bleeding was 3. 4% in the research group and 6. 7% in the control with no statistically significant difference (χ2 =0. 316,P >0. 05), but there was no mechanical injury of oral mucosa in two groups. There was no statistically significant difference in index of water-swallowing test between two groups before interventions ( P>0. 05), with significant difference at 4th week (P<0. 01). Conclusions The implementation of mouth nursing by modified method of brushing their teeth and irrigation not only can achieve the purpose to clean oral effectively, but also can reduce oral bacteria and promote the recovery of swallowing function.