Objective To develop prototype foamy virus (PFV) detection method by loop-mediated isothermal amplification. Methods Three pairs of primers targeting core region of PFV integrase were designed in this study and Bst DNA polymerase was used to amplify target sequence at 63℃. The system was established with all the conditions optimized. Results The method was established with the plasmid containing target sequence as the template. This method could specifically detect PFV infectious clone, no crossreaction was observed with human immunodeficieney virus infectious clone, bovine immunodefieiency virus infectious clone and bovine foamy virus infectious clone as templates. The detection capability of this system was 50 copy, one order more sensitive than PCR. The amplification could be finished in 15 min and human genomic DNA did not adversely affect the amplification efficiency. Conclusion The PFV detection method by loop-mediated isothermal amplification was established and it had potential usefulness in PFV detection.

ISG15, the first ubiquitin-like molecule identified two decades ago, is encoded by interferon stimulated gene 15 ( ISG 15), where its robust expression can be induced by viral infections or interferon treatments. ISG 15 conjugate to other proteins as the ubiquitin and was found to be involved in innate immune response. However, the functions of ISG15 modification remained unclear. We cloned bovine ISG15(bisg15) into a prokaryotic expression vector pET28a( + ) with a His-tag to generate a soluble form of bISG15 fusion protein, and purified with Ni-NTA Sepharose chromatography. The purified protein was concentrated and used to immune Balb/c mice to raise the antiserum, which could specifically recognize bISG15 expressed in eukaryotic cells by Western blot analysis. The concentrated bISG15 protein and its antiserum were then used to establish an in vitro bISG15 modification system. Our studies have demonstrated that cellular proteins could be conjugated to bISG15 with this system.

The ubiquitin-like modifier bISG15 is an antiviral protein found in fetal bovine lung (FBL) cells.Bovine Herpesvirus 1(BHV-1),which is a viral pathogen of cattle,can infect FBL cells and induce cytopathic effects.Real-time PCR assays showed that BHV- 1 's infection could repress the basal or inducible transcription of bISG15 in FBL cells.It demonstrates that this repression effect depends on BHV-1 viral infection and new protein synthesis.Our previous work showed that bIRF-3 was the key factor in the stimulation of bISG 15 in FBL cells,so the effect of BHV-1 viral protein on bIRF-3 activating the promoter of bISG15 was confirmed.The luciferase assay showed the BHV-1 viral protein bICP0 inhibited the activation of bISG15 promoter stimulated by bIRF-3.Taken together,our work suggested that BHV-I had some molecular mechanism to resist the cellular bISG15'santiviral functions.

OBJECTIVETo investigate the influence of male age on the pregnancy outcomes of in vitro fertilization-embryo transfer (IVF-ET).

METHODSWe retrospectively analyzed 7,533 cycles of IVF-ET performed between January 1, 2009 and October 31, 2013. We divided the samples into three groups according to the female age (< 30, 30-34, and 35-38 yr), each again subdivided into six groups based on the male age (< 30, 30-32, 33-35, 36-38, 39-41, and ≥ 42 yr). We compared the rates of implantation, pregnancy, miscarriage, and live birth among different age groups.

RESULTSThere were no statistically significant differences in basal E2, FSH, endometrium thickness on the day of hCG administration, number of oocytes retrieved, and days of embryo transfer among different male age groups (P > 0.05). The implantation rate showed an age-dependent decrease in the < 30, 30-32, 33-35, 36-38, 39-41, and ≥ 42 yr male groups, 41.1, 42.0, 39.5, 31.3, 40.7, and 48.6% among the women aged < 30 years (P < 0.05), 40.3, 36.4, 35.1, 35.3, 29.4, and 37.3% among the women aged 30-34 years (P < 0.05), and 48.2, 17.8, 25.3, 23.5, 22.1, and 23.8% among the women aged 35-38 years (P < 0.05). The miscarriage rate was significantly higher in the ≥ 39 yr than in the 30-32 and 33-35 yr male age groups among the women aged 30-34 years (P < 0.05), but showed no remarkable differences among the other male age groups in the women aged < 30 and 35-38 years (P > 0.05). No statistically significant differences were observed in the rates of pregnancy and live birth among different male age groups (P > 0.05).

CONCLUSIONMale age has some influence on the rates of implantation and miscarriage but not on the rates of pregnancy and live birth in IVF-ET.

Abortion, Spontaneous ; epidemiology ; Adult ; Age Factors ; Embryo Implantation ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Male ; Oocyte Retrieval ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Retrospective Studies ; Sex Factors

Animals ; Base Sequence ; Gene Expression Regulation ; Humans ; MicroRNAs ; genetics ; metabolism ; Molecular Sequence Data ; RNA, Viral ; genetics ; Virus Diseases ; genetics ; metabolism ; virology ; Virus Physiological Phenomena ; Viruses ; genetics ; metabolism

Objective To evaluate the role of high-risk human papillomavirus (HR-HPV) testing in diagnosis of high-grade cervical intraepithelial neoplasia (CIN2/3+) and cervical cancer.Methods Total 3426 women with known cytologic abnormality tests were subjected to detect high-risk HPV DNA by hybrid capture II and to have established diagnoses by cervical coloposcopy-directed biopsy test.Results Among 3426 women,45 were cervical cancer,670 were CIN 2/3,and 2711 were CIN 1 or other benign lesions based on the cervical histologic results. Compared with the women with CIN 1 or other benign lesions,the women with CIN 2/3+ had higher positive rates of HR-HPV. Higher load of HR-HPV was more common among women with cervical lesions than those without. Fifty-seven women who were transiently HR-HPV positive did not develop CIN2/3+ during follow-up.Conclusions Our data suggest that a high viral load could be used as a short-term marker of CIN 2/3+,and women repeatedly tested positive for HR-HPV are at risk of developing CIN2/3+. Detecting HR-HPV and follow up the positive women would be useful in diagnosis of CIN2/3+.

Virus infection or interferon can stimulate robust expression of the protein ISG15 that is encoded by interferon stimulated gene 15, which was the first unbiquitin-like molecule identified two decades ago. While ubiquitin and its many important functions have been well established, the functions of ISG15 and its post-translational conjugation are still largely unknown . Recently, some specific enzymes have been identified to be involved in the ISG15 modification system, suggests that ISG15 and its modification system play important roles in the innate immune response and regulation of interferon signaling. The history of ISG15 discovery and its biochemical characterization were briefly introduced. Then such topics as the ISG15 gene expression and the ISG15 modification will be focued on, and finally summarize new findings which have implications for ISG15 and its modification system in immunology and interferon signal transduction were summarized.

Objective:To detect the clinical features of vulvar and vaginal intraepithelial neoplasias(VIN and VAIN,respectively).Methods:Total 148 women were performed vulvar or vaginal coloposcopy-directed biopsy pathology tests,from Sep.2004 to Dec.2007.Results:Among 148 women,vulvar or vaginal histologic results were vulvar cancer for 1,VIN or VAIN 2,3 for 23,VIN or VAIN for 16,condyloma for 61,vulvitis and vaginitis for 47.Eighty-five percent(33/39) women with VIN or VAIN 2,3 were more than 30 years old.Compared to women with cervical intraepithelial neoplasia(CIN),women with VIN or VAIN were older.The rate of high-risk HPV DNA in women with vulvar or vaginal lesions was 84%(84/100).VAIN occurred mainly in the upper vagina(90%,69/75).VIN or VAIN often accompanied or followed CIN or cervical cancer(79%,31/39),and VIN or VAIN 2,3 often accompanied or followed CIN 2,3 or cervical cancer(70%,16/23).Conclusion:Our data suggest that women with high-risk HPV infection are at risk of developing VIN or VAIN 2,3.The vulva and vagina should be carefully inspected by colposcopic examination at the time of colposcopy for any abnormal findings.

Objective To summarize the clinical features of different racial patients with celiac disease (CD) and analyze the disease prevalence,diagnosis and treatment in Chinese population.Methods All the patients were diagnosed as CD and enrolled in Beijing United Family Hospital between January 2005 and July 2015.Clinical data including nationality,age,symptoms,endoscopic and pathological findings,outcome were collected and compared in patients from different countries.Results A total of 87 patients were enrolled including 63 Caucasians,18 Asian patients and 6 Middle East patients.The peak age of disease onset was 40-60 years old.Patients with typical symptoms such as chronic diarrhea and weight loss only accounted for 20.7％ (18/87) and 9.2％ (8/87) respectively.Some patients presented with nonspecific symptoms such as abdominal pain and bloating [32.2％ (28/87)],even constipation [5.7％ (5/87)].13.8％ (12/87) patients were previously diagnosed as irritable bowel syndrome.The incidence of abdominal pain,bloating,diarrhea and constipation between Asians and Caucasians had no statistical significance (P ＞ 0.05);but the proportions of weight loss,growth retardation,iron deficiency anemia and dermatitis herpetiformis in Asian group were significantly higher than that in Caucasian group (P ＜ 0.05).IgA type of anti-gliadin antibody (AGA),endomysium antibody (EMA) and tissue transglutaminase antibody (tTGA) were dominant autoimmune antibodies in patients with CD,which accounted for 58.6％ (51/87),44.8％ (39/87) and 36.8％ (32/87) respectively.The endoscopy showed that the lesion of CD was mainly located in small intestine,with reducing severity from the proximal to the distal small intestine.The lesions of duodenal bulb and descending duodenum appeared more significant in Asian group.Accordingly pathological intestinal atrophy and the degree of intraepithelial lymphocytosis were more severe in Asian patients.All 87 cases took the gluten-free diet (GFD).Eighty-one cases received serological follow up and 8 with endoscopic intestinal biopsy.The celiac disease antibodies in 47 patients turned negative from 6-9 months after GFD treatment,while 34 patients turned negative from 12-18 months after GFD.All patients reported disease remission to some extent.After 1 year GFD treatment,the pathology of endoscopic intestinal biopsy in 8 patients showed significant improvement of villous atrophy and lymphocyte infiltration.Conclusions CD patients with typical clinical manifestations are not the majority.Serological celiac disease antibodies (AGA,EMA and tTGA) have a high diagnostic value.GFD treatment is effective on majority of celiac patients.Clinical manifestations,endoscopy,intestinal pathology,and response to GFD in Chinese patients are not the same as Caucasians.Clinicians need to pay attention to the differential diagnosis.