[Effects and mechanism of lncRNA serving as ceRNA in non-obstructive azoospermia].

Objective: To investigate the mechanisms of lncRNA on the occurrence and development of NOA by constructing ceRNA regulation network of lncRNA, miRNA and mRNA. Methods: Samples of adult human testis were obtained from NOA patients and OA patients with normal spermatogenesis (controls), recruited from the Reproductive Medicine Center of Nanfang Hospital from June 2017 to June 2018. Differentially expressed lncRNAs and mRNAs in testicular tissues from patients with NOA were identified by microarray analysis in previous association study. In this study, differentially expressed lncRNAs and mRNA were used to construct the ceRNA regulatory network in NOA and clarify the interaction relationship among lncRNA, miRNA and mRNA. GeneMANIA database was used to construct Protein-Protein Interaction (PPI) of the mRNAs in ceRNA regulatory network. WebGestalt toolkit was employed to perform gene function and pathway enrichment analyses of those coding genes. Finally, qRT-PCR and dual luciferase reporter system were employed for further experimental validation. Results: The ceRNA regulatory network of lncRNA, miRNA and mRNA consists of 21 nodes and 26 edges, of which 4 lncRNAs, 13 miRNAs and 4 mRNAs. 19 proteins were found to interact with the mRNA coding proteins in ceRNA regulatory network by PPI analysis. Gene oncology and KEGG pathway enrichment analyses indicate these coding genes were significantly enriched in pentose metabolic process and pentose phosphate pathway. Furthermore, lncRNA ANXA2P3 was found binding with miR-613 and miR-206 to inhibit mRNA TKT expression. Conclusion: lncRNAs exert an important role in the occurrence and development of NOA via ceRNA regulatory network, which could be used as new biomarkers for NOA treatment.

Polymorphisms of KAP6, KAP7, and KAP8 genes in four Chinese sheep breeds.

High glycine-tyrosine proteins (HGTPs), also known as keratin-associated proteins (KAPs), play a key role in the major structures and mechanical properties of wool fiber. Sheep HGTPs consist of three multigene families: KAP6, KAP7, and KAP8 genes. Polymorphisms of these three genes have been proposed to have important effects on wool fiber traits. The aim of the present study was to identify polymorphisms of the KAP6, KAP7, and KAP8 genes in four sheep breeds, including Chinese Merino superfine wool sheep, Hu sheep, a Merino x Hu crossed breed, and Romney sheep. Polymerase chain reaction (PCR) product direct sequencing, PCR-single-strand conformation polymorphism, and cloned sequencing methods were used to find genetic variation and identify polymorphisms in these genes. The Mutation Surveyor v3.97 software was used to analyze the sequences. These methods revealed six different sequences of the KAP6 gene, two different sequences of the KAP7 gene, and five different sequences of the KAP8 gene. Accordingly, three (with frequencies>1%) single nucleotide polymorphisms (SNPs) of the KAP6 gene, one SNP of the KAP7 gene, and five SNPs of the KAP8 gene were detected. Interestingly, some of these sequences were present in only certain sheep breeds, thereby suggesting that these special allele sequences could be used as candidate genes of wool characteristics in further studies.

αv ß3 Integrin may participate in conceptus attachment by regulating morphologic changes in the endometrium during peri-implantation in ovine.

The objective of this study was to determine expression and potential functions of α(v) and ß(3) integrin subunits in ovine endometrium during the peri-implantation period (days 8-17 after fertilization). The morphologic changes in the endometrium were observed histochemically following haematoxylin/eosin (HE) staining, whereas the expressions of α(v) and ß(3) integrin subunits were analysed by RT-PCR, immunohistochemistry and Western blot. The filamentous conceptus attached to the luminal epithelium (LE) on day 17 of pregnancy, with no differences in endometrial morphology between days 8-12 of pregnancy. However, endometrial glands in the endometrial stroma (S) underwent extensive hyperplasia from day 14 to day 17, increased reductus of the LE with an obvious proliferation of caruncles, and an increased number and diameter of blood vessels (V) in the endometrium. The relative expression levels of α(v) and ß(3) integrin subunits mRNA gradually increased until day 16, but sharply declined on day 17. Western blot analysis revealed that the expression pattern of α(v) and ß(3) integrin subunit proteins paralleled that of the corresponding mRNA. In addition, immunohistochemical localization of α(v) and ß(3) integrin subunits confirmed their presence in the glandular epithelium (GE), LE and endometrial stroma. Immunostaining on LE and stroma varied with the increasing days of pregnancy, with the strongest immunostaining on days 16 and 17. In conclusion, expression of α(V) and ß(3) integrin subunits was closely related to the early progression of pregnancy and conceptus attachment; therefore, we inferred that α(v) ß(3) integrin may participate in conceptus attachment by the regulation of endometrial morphology during peri-implantation in ovine.

Up-regulation of expression of interferon-stimulated gene 15 in the bovine corpus luteum during early pregnancy.

Interferon-tau (IFNT), the pregnancy recognition signal in ruminant species, is secreted by conceptus trophectoderm cells and induces expression of IFN-stimulated gene 15 (ISG15) in the uterus and corpus luteum (CL) in ewes. Expression of ISG15 in ovine CL is speculated to be through an endocrine pathway, but it is unclear whether expression of ISG15 in bovine CL is via such a pathway. In this study, CL were obtained from cows on d 16, 25, 60, 120, 180, and 270 of pregnancy, and endometrium, mammary gland, ovarian stroma, and CL were also collected from cows on d 18 of pregnancy and on d 15 and 18 of the estrous cycle. All tissue explants from d 15 of the estrous cycle were cultured in the absence or presence of 100ng/mL of recombinant bovine IFNT for 24h. The results indicated that ISG15 and conjugated proteins were expressed in CL of both cyclic and pregnant cows regardless of pregnancy status and were upregulated during early pregnancy. The mammary gland from d 18 of pregnancy did not express ISG15, but explants of the mammary gland from d 15 of the estrous cycle did express ISG15 after being treated with IFNT. However, luteal explants from d 15 of the estrous cycle did not express ISG15 after being cultured for 24h. In conclusion, ISG15 expression is upregulated in the bovine CL during early pregnancy. Interestingly, cultured CL cells do not respond to IFNT, suggesting that the pregnancy-dependent stimulation of ISG15 expression is controlled by something other than IFNT in the bloodstream.

Expression of interferon-tau mRNA in bovine embryos derived from different procedures.

Interferon-tau (IFN-tau) is a secreted conceptus protein which plays a critical role in the establishment of ruminant pregnancy by its antiluteolytic and antiviral effects. In the present study, we hypothesized that IFN-tau expression was temporally and spatially regulated in different pre-implantation embryos and the levels of IFN-tau expression were different among bovine embryos derived from parthenogenetic activation (PA), in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT). By using in situ hybridization with Digoxingenin (DIG)-labelled IFN-tau cDNA as a probe, we detected IFN-tau mRNA in bovine embryos from days 3 to 9 in culture. However, the timing of the initiation of IFN-tau mRNA expression was different among PA, IVF and SCNT embryos. Interferon-tau mRNA was first expressed in 16-cell stage IVF embryos on day 4, in SCNT morula on day 5 and early PA blastocyst on day 6. Semi-quantitative RT-PCR analysis showed that the expression levels of IFN-tau mRNA did not differ significantly among IVF, SCNT and PA embryos on day 7. In addition, freezing and thawing did not have a major impact either on IFN-tau mRNA expression in IVF or in vivo-produced bovine blastocysts.

Aspergillosis among people infected with human immunodeficiency virus: incidence and survival. Adult and Adolescent Spectrum of HIV Disease Project.

Aspergillosis is a life-threatening fungal infection in immunocompromised people, including people infected with human immunodeficiency virus (HIV). We determined the incidence of aspergillosis among HIV-infected people and survival after aspergillosis diagnosis by use of a national HIV surveillance database. Among 35,252 HIV-infected patients, the incidence of aspergillosis was 3.5 cases per 1000 person-years (p-y; 95% confidence interval [CI], 3.0-4.0 per 1000 p-y). Incidence was higher among people aged > or =35 years (4.1 per 1000 p-y, 95% CI, 3. 5-4.8), among people with CD4 counts of 50-99 cells/mm(3) (5.1 per 1000 p-y, 95% CI, 2.8-7.3), or CD4 counts of <50 cells/mm(3) (10.2 per 1000 p-y, 95% CI, 8.0-12.2), versus people with CD4 counts of >200 cells/mm(3), people with > or =1 acquired immune deficiency syndrome-defining opportunistic illness (8.6 per 1000 p-y, 95% CI, 7.4-9.9), and people who were prescribed at least one medication associated with neutropenia (27.7 per 1000 p-y, 95% CI, 21.0-34.3). Median survival time after diagnosis of aspergillosis was 3 months, and 26% survived for > or =1 year. These findings suggest that aspergillosis is uncommon, occurs especially among severely immunosuppressed or leukopenic HIV-infected people, and is associated with poor survival.

Effect of hormone replacement therapy on breast cancer risk: estrogen versus estrogen plus progestin.

BACKGROUND: Hormone replacement therapy (HRT) given as unopposed estrogen replacement therapy (ERT) gained widespread popularity in the United States in the 1960s and 1970s. Recent prescribing practices have favored combination HRT (CHRT), i.e., adding a progestin to estrogen for the entire monthly cycle (continuous combined replacement therapy [CCRT]) or a part of the cycle (sequential estrogen plus progestin therapy [SEPRT]). Few data exist on the association between CHRT and breast cancer risk. We determined the effects of CHRT on a woman's risk of developing breast cancer in a population-based, case-control study. METHODS: Case subjects included those with incident breast cancers diagnosed over 4(1/2) years in Los Angeles County, CA, in the late 1980s and 1990s. Control subjects were neighborhood residents who were individually matched to case subjects on age and race. Case subjects and control subjects were interviewed in person to collect information on known breast cancer risk factors as well as on HRT use. Information on 1897 postmenopausal case subjects and on 1637 postmenopausal control subjects aged 55-72 years who had not undergone a simple hysterectomy was analyzed. Breast cancer risks associated with the various types of HRT were estimated as odds ratios (ORs) after adjusting simultaneously for the different forms of HRT and for known risk factors of breast cancer. All P values are two-sided. RESULTS: HRT was associated with a 10% higher breast cancer risk for each 5 years of use (OR(5) = 1.10; 95% confidence interval [CI] = 1.02-1.18). Risk was substantially higher for CHRT use (OR(5) = 1.24; 95% CI = 1.07-1.45) than for ERT use (OR(5) = 1. 06; 95% CI = 0.97-1.15). Risk estimates were higher for SEPRT (OR(5) = 1.38; 95% CI = 1.13-1.68) than for CCRT (OR(5) = 1.09; 95% CI = 0. 88-1.35), but this difference was not statistically significant. CONCLUSIONS: This study provides strong evidence that the addition of a progestin to HRT enhances markedly the risk of breast cancer relative to estrogen use alone. These findings have important implications for the risk-benefit equation for HRT in women using CHRT.

Progressive multifocal leukoencephalopathy: improved survival of human immunodeficiency virus-infected patients in the protease inhibitor era.

To examine factors affecting survival after diagnosis of progressive multifocal leukoencephalopathy (PML), we analyzed data from an observational cohort study, the Adult and Adolescent Spectrum of HIV Disease project. We identified 415 patients diagnosed with PML during 1990-1997. The median survival time after diagnosis was 1 month. By use of an extended proportional hazards, multivariate regression model, risk factors associated with decreased survival time included CD4 count <0.20 x 10(9) cells/L (risk ratio [RR], 2.1; 95% confidence interval [CI], 1.3-3.5) compared with >/=0.20 x 10(9) cells/L, whereas factors associated with increased survival time were prescription of antiretroviral medication that contained a protease inhibitor (RR, 0.2; 95% CI, 0.1-0.4) and prescription of other antiretroviral medication (RR, 0.6; 95% CI, 0.5-0.8) compared with no antiretroviral prescription. We conclude that protease inhibitor use (in combination antiretroviral therapy) is likely to favorably affect survival time after diagnosis of PML.

Trends in zidovudine prescription for pregnant women infected with HIV.

OBJECTIVE: The purpose of this analysis was to describe trends in zidovudine prescription during pregnancy among women infected with HIV. METHODS: We used data from the Adult and Adolescent Spectrum of Disease Surveillance Project, which collects information on HIV-related conditions through medical record review. Women who were reported pregnant from 1990 through 1996 were included in the analysis. RESULTS: From 1990 through 1996, of 7047 women in the project, 714 (10%) were pregnant for a total of 782 pregnancies. We found a high proportion (82%) of pregnancies during which zidovudine was prescribed for women with CD4+ T-lymphocyte count of 0 to 199 cells/microl (n = 125), but no trend over time. In contrast, from 1990 through 1996 zidovudine was prescribed for an increasing proportion of pregnancies in which the woman's CD4+ count was 200 to 499 cells/microl (62%-78%; p = .01; n = 337) and > or = 500 cells/microl (22%-55%; p = .001; n = 250). CONCLUSIONS: Our study demonstrated differences in zidovudine prescription over time by CD4+ count; these differences may be based on the woman's health and guidelines for perinatal prevention.

Estrogen-progestin replacement therapy and endometrial cancer.

BACKGROUND: It has been known for more than 20 years that estrogen replacement therapy substantially increases a woman's risk of developing endometrial cancer. To reduce this increased risk, progestins have been added to estrogen replacement therapy for between 5 and 15 days (usually 7 or 10 days) per "month" in a sequential fashion (sequential estrogen-progestin replacement therapy) or with each dose of estrogen replacement therapy (continuous combined replacement therapy). At the present time, however, little is known about the effects of varying the number of days that progestin is used in sequential estrogen-progestin replacement therapy. PURPOSE: We sought to determine the effects of sequential estrogen-progestin replacement therapy and continuous combined replacement therapy on a woman's risk of developing endometrial cancer. METHODS: A population-based, case-control study of 833 case subjects and 791 control subjects was conducted. Women were postmenopausal, white, and aged 50-74 years when first diagnosed with invasive endometrial cancer or were aged 50-74 years at the matching date for control subjects. All subjects were interviewed in person with the aid of a month-by-month calendar. Relative risks were estimated by odds ratios (ORs); ORs were adjusted simultaneously for the different forms of hormone replacement therapy and for the known endometrial cancer risk factors. RESULTS: The adjusted OR was 2.17 (95% confidence interval [CI] = 1.91-2.47) per 5 years of estrogen replacement therapy use (based on 422 users among the case subjects and 262 users among the control subjects). For women who received sequential estrogen-progestin replacement therapy with the progestin given for less than 10 days (effectively 7 days) per month, the adjusted OR was only slightly reduced to 1.87 (95% CI = 1.32-2.65) per 5 years of use (74 case subjects and 47 control subjects). However, when progestin was given for 10 or more days (effectively 10 days), there was essentially no increased risk (adjusted OR = 1.07 per 5 years of use; 95% CI = 0.82-1.41) (79 case subjects and 88 control subjects). Continuous combined replacement therapy was also associated with essentially no increased risk (adjusted OR = 1.07 per 5 years of use; 95% CI = 0.80-1.43) (94 case subjects and 81 control subjects). CONCLUSIONS: The progestin in sequential estrogen-progestin replacement therapy needs to be given for at least 10 days to block effectively any increased risk of endometrial cancer. Continuous combined estrogen-progestin therapy is similarly effective. Neither regimen reduces a woman's underlying risk of endometrial cancer. The sharp distinction between the effects of less than 10 days (effectively 7 days) and 10 or more days (effectively 10 days) of progestin use in sequential estrogen-progestin replacement therapy suggests that the extent of endometrial sloughing may play a critical role in determining endometrial cancer risk.

An exchange of ophthalmologists with the People's Republic of China.

From 1978 through 1981, three teams of ophthalmologists from the United States have traveled to the People's Republic of China under the auspices of the World Eye Foundation. The purpose of these visits has been to open an avenue of exchange between American and Chinese ophthalmologists. The US ophthalmologists delivered lectures and participated in discussion groups with the Chinese ophthalmologists. They had the opportunity to participate in surgical teaching cases, including some using acupuncture anesthesia. They also had the opportunity to develop ongoing working and research relationships with many Chinese ophthalmologists. As a result of these working relationships, some of these Chinese ophthalmologists have had the opportunity to visit and study in the United States.