AIM:Antioxidants act mainly through two routes:induction of antioxidant enzymes(enzymatic)or direct reaction with reactive oxygen species(ROS)(non-enzymatic),but the one taken by the extract of Ginkgo biloba(EGb)remains to be investigated.In present study,EGb was tested for both of the antioxidant routes in vitro.METHODS:The induction of EGb on two antioxidant enzymes,glutamate cysteine ligase catalytic subunit (GCLC)and glutathione-S-transferage subunit-P1(GST-P1),in cell lines wag detected by Western-blot.The effects of EGb on superoxide anion radical(O2·-),hydroxyl radical(OH·),rat erythrocyte hemolysis and lipid peroxidation of rat liver homogenate were determined by activity methods respectively.RESULTS:EGb was dem onstrated significantly to induce the two antioxidant enzymes(GCLC and GST-P1),directly scavenge superoxide anion radical(O2·-),hydroxyl radical(OH·)and inhibit rat erythroeyte hemolysis and lipid peroxidation of rat liver homogenate.CONCLUSION:The results strongly support the notion that EGb plays dual antioxidant roles: induction of antioxidant enzymes and direct reaction with ROS.

Objective To clone the gene of arginine kinase (AK) from Periplaneta americana, produce its recombinant protein and investigate its allergenicity. Method The cDNA of AK was cloned using specific primers from the total RNA of P. americana. The cloned gene was inserted into pMD18-T vector and digested by BamHI and HindⅢ. The cDNA was sequenced and subcloned into pET-28a expression vector. The cloned AK cDNA gene was expressed in Escherichia coli BL21 (DE3) by IPTG induction. The recombinant AK (rAK) was purified by metal (Ni2+) chelating affinity chromatography. Its allergenicity was examined by both Western blotting and enzyme-linked immunosorbent assay (ELISA). Result The cloned cDNA ORF sequence (Accession no. EU429466) contained 1 068 bp and encoded 365 amino acids. Its sequence homology with the published one (Accession no. AY563004) was 99.9% at nucleotide level. The allergen rAK was highly expressed in E. coli BL21 (DE3) as a soluble protein mainly with the molecular weight of about Mr 45 000 under induction of IPTG and purified by 6-His-tag purification system. Both in the non-denaturalization and denaturalization conditions, the recombinant allergen was identified as its affinity to IgE antibodies from the cockroach-allergic patient sera by Western blotting and ELISA. Conclusion The recombinant cockroach arginine kinase has been obtained with proper allergenicity.

Objective:To investigate effects of over-expression and suppression of HMGB1 on proliferation and invasion of endometrial carcinoma HEC-1A cell and underlying mechanisms.Methods:Over-expression or silence of HMGB 1 in HEC-1A cell lines were established by lentiviral vector containing HMGB 1 recombinant plasmid or by HMGB 1 shRNA,respectively.Cell counting kit-8,Transwell,and wound healing assay were used to analyze proliferation,invasion,and migration of HEC-1A cells,respectively.Western blot and reverse transcription-PCR (RT-PCR) were used to detect the expression of NF-κB,VEGF,and matrix metalloproteinase 2 (MMP2) in the cells.Results:Over-expression of HMGB1 promoted the proliferation,invasion,and migration of HEC1A cell,and up-regulated NF-κB,VEGF,and MMP2 expressions,while suppression of HMGB1 inhibited the proliferation,invasion,and migration of HEC-1A cells and down-regulated NF-κB,VEGF,and MMP2 expressions.Conclusion:HMGB1 plays an important role in proliferation,invasion,and migration of HEC1A cell via NF-κB/VEGF/MMP2 pathway.HMGB 1 might be a potential target for endometrial carcinoma therapy.

Objective To explore the association of anti-mullerian hormone(AMH)in seminal plasma and serum with sperm counts and energy for male.Methods For 215 cases of healthy male selected from our reproductive clinic,with women′s reason for infertility,seminal plasma and serum AMH were detected,as semen parameters(sperm density,living rate,vitality and malformation rate),6 items of serum sex hormone.In seminal plasma and serum AMH respectively as the dependent variable,using multiple line-ar regression model to explore its quantitative relation with semen parameters and sex hormone levels.Results 215 cases were en-rolled,aged 34.28±5.70 years,while the median of the seminal plasma AMH was 0.47,quartile 0.05-3.09 pmol/ejaculation.The median of the serum AMH was 53.07,quartile 32.32 -72.20 pmol/L.Through multiple linear regression analysis,after adjusted by age and BMI,the seminal plasma of AMH and total number of sperm,sperm concentration,dynamic motility,total sperm activi-ty,serum inhibin B were positively correlated(P <0.05);The correlation between sperm morphology and other serum sex hormone had no statistical significance(P >0.05);Serum AMH negatively correlated with serum FSH,with serum inhibin B positively(P <0.05);Seminal plasma in various parameters and other related serum sex hormone had no statistical significance(P >0.05).Conclu-sion The seminal plasma of AMH were positively correlated with sperm concentration,sperm counts,sperm vitality,with the asso-ciation for serum AMH not yet found.

Objective To investigate the knowledge level and needs in Xinjiang Uygur and Han patients with ulcerative colitis (UC), compare the differences between the two ethnic groups. Methods A total of 194 Uygur and Han UC patients were investigated with the general information questionnaire, the Crohn′s and Colitis Knowledge Score (CCKNOW) questionnaire, disease related knowledge needs questionnaire, to analyze the investigate results. Results CCKNOW score of Uygur and Han UC were (6.9±3.5) points and (9.2±3.1) points respectively, and Uygur was significantly lower than that of Han (Z=-2.831, P=0.005). Knowledge accuracy of dietary (30% and 63%) and drug (45% and 44%) were higher than general information (24% and 32%) and complications (12% and 30%) . Disease related knowledge needs score of Uygur and Han UC patients were (168.2±15.6) points and (155.4±17.2) points respectively, the score of Uygur patients was significantly higher than that of Han patients (t=4.429, P=0.001).The highest disease related knowledge needs score was (4.7±1.0) points of reproductive knowledge for Uygur UC patients, for Han patients was knowledge of daily life which was (4.8 ± 0.8) points. Conclusions The disease related knowledge level of Xinjiang Uygur and Han UC patients are relatively low, especially lack of general knowledge and diet knowledge, disease related knowledge of Uygur UC patients are lower than Han patients. The needs of the disease knowledge are very high for two ethnic groups, different one has different ethnic, gender, age, education level, living environment, hospitalization times, areas of disease related knowledge needs are also different. It is necessary to choose the targeted education content and suitable education way according to individual differences for nursing staff.

AIM To modify and improve a screening assay so that it becomes more convenient, economic and adaptable in China for high throughput screening of HIV fusion inhibitors targeting gp41. METHODS The original screening method reported by Jiang et al (J Virol. Methods 1999;80:85 96) was modified by: ① using a conformation specific monoclonal antibody to replace a polyclonal antibody for coating plates; ②simplifying the procedures; ③using parts of the reagents produced in China. RESULTS The modified screening assay is simpler, more convenient, and more economic than the original assay, but its sensitivity is comparable to and specificity is a little better than the original method. CONCLUSIONS The modified screening assay is more convenient and economic and can be used in China for high throughput screening of HIV fusion inhibitors from complex sample, such as phage display peptide libraries, microorganism fermentation liquids, herbs and other natural products.

Objective To study the effects of folate(Fol) on proliferation,apoptosis and the antioxidative capacity of the NSCs under hypoxia in vitro.Method The NSCs taken from infant rats brain were cultured by serum-free medium in vitro and divided into four group which were normal control group(NC),hypoxia group(Hyp),Hpy+Fol-D(folate deficient)group and Hpy+Fol group.Except NC group,the other groups were cultured in hypoxia condition for 6 h.The proliferation of NSCs after hypoxia damage was detected by MTT test.The NSCs were collected after being cultured 6 d and then the activity of SOD and GSH-PX was examined.The expression of activated caspase-3 was detected by Western blot.Results Compared with NC group,the proliferation of NSCs and the activity of SOD and GSH-PX decreased significantly after hypoxia,while the expression of activated caspase-3 increased0 The proliferation of NSCs and the activity of SOD and GSH-PX in Hyp group were significantly higher than Hyp+Fol-D group but much lower than Hpy+Fol group(P

Objective To investigate the efficacy and tolerability of estradiol (E2) gel with postoperative patients of moderately severe ovarian endometriosis cyst,who suffer from the side effect of GnRH agonist.Methods Sixty samples were selected with stage Ⅲ ～ Ⅳ ovarian endometriosis after conservative operation by the same operator at the Department of Gynecology,Xiangya Hospital,Central South University,between February 2013 and August 2014.All of the patients after surgeries were randomly divided into three groups:control group (n =20) with only goserelin injection after conservative surgery,treatment group A (n =20) who received estradiol gel,and treatment group B (n =20) who received tibolone as an add-back therapy.The serum E2,follicle stimulating hormone (FSH),luteinizing hormone (LH),hypoestrogenic symptoms,the incidence of uterine bleeding,and the recurrence after conservative operation were compared among three groups.Results Three months later aster treatment with goserelin,the serum level of E2 of three groups showed statistically significant difference (P ＜ 0.05).Serum E2 level of treatment group A was not significantly higher than treatment group B (P ＞ 0.05).The kupperman sore,the incidence of hot flashes sweating,tiring,and agrypnia of treatment groups A and B were significantly lower than the control group (P ＜ 0.05).The incidence of uterine bleeding and recurrence dont differ sharply among three groups (P ＞ 0.05).Conclusions GnRHa combined with estradiol gel can improve symptoms of postoperative patients,reduce side effects effectively,and avoid increase of the risk of vagina bleeding and relapse.

Objective To investigate the association between high-risk types of human papillomavirus (HR-HPV) load and the expression of nestin in different cervical lesions.Methods The hybridization capture Ⅱ (HC-Ⅱ) assay was used to test the HR-HPV load from 60 patients who were the first time to do the cervical cancer screening in Xiangya Hospital,and immunohistochemistry was used to detect the expression of nestin in those biopsy tissue samples.Results (1) The lgHPV level (logarithm of HR-HPV load) in the high level lesion group was higher than the low level one (P ＜0.05),and HR-HPV load was positively associated with the degree of cervical lesions (rs =0.269,P =0.037).(2) The expression of nestin in A group was weaker than groups B and C (H =7.271,22.843,P ＜0.01),and the expression of nestin in C group was stronger than B group (H =7.270,P ＜0.01),and the expression of nestin was positively associated with the degree of cervical lesion (rs =0.646,P =0.000).(3) The HR-HPV load was positively associated with the expression of nestin (P ＜ 0.05).Conclusions The HR-HPV load and the expression of nestin are closely related to the cervical lesions,and the joint detection has a referential value for early prevention of cervical lesions and prediction of progress of cervical precancerous lesion.It might guide the early prevention and treatment of cervical cancers.

BACKGROUND: There are currently many cryopreservation methods for the aminotic membrane, which have varying effects on the ultrastructure and biological activity of amniotic membrane, but on no one is effective.
OBJECTIVE: To compare the effects of different cryopreservation methods on the ultrastructure and viability of aminotic membrane and to seek the ideal cryopreservation method.
METHODS: Aminotic membrane separated from the fresh placenta was preserved respectively with deep-frozen cryopreservation and vitrification, and everyway was run for 3 and 6 months. Fresh aminotic membrane was used as control. The ultrastructure of aminotic membrane was observed by transmission electron microscopy, and the viability of aminotic membrane was assessed by microcomputer analysis system for biological oxygen consumption, and immunohistochemical staining combined with image analysis system was used for lactate dehydrogenase activity.
RESULTS AND CONCLUSION:After 3 and 6 months of crypreservation, the damage to the ultrastructure of aminotic membrane by vitreous cryopreservation was slighter than that of amniotic membrane cryopreserved at-80℃. Compared with the fresh aminotic membrane, the gray value of lactate dehydrogenase and partial pressure of oxygen were significantly decreased in the cryopreserved aminotic membrane by deep-frozen cryopreservation at 3 and 6 months (P < 0.05) and by vitreous cryopreservation at 6 months (P < 0.05), but there was no statisticaly significant difference in the change rate of oxygen partial pressure and the gray value of lactate dehydrogenase between the fresh aminotic membrane and the cryopreserved aminotic membrane by vitreous cryopreservation at 3 months. The present study led to the conclusion that vitreous cryopreservation protocol alows to not only maintain the integrity of AM, but also to preserve the viability of the cels. So the vitreous cryopreservation is superior to the deep-frozen cryopreservation for cryopreservation of aminotic membrane.