VDR gene ApaI polymorphism and risk of postmenopausal osteoporosis: a meta-analysis from 22 studies.

OBJECTIVE: The ApaI polymorphism (G > T, rs7975232) of the vitamin D receptor (VDR) gene in the risk of postmenopausal osteoporosis has been widely researched, and the results have yielded conflicts. Therefore, we performed an updated pooled analysis to comprehensively assess the association between VDR ApaI polymorphism and postmenopausal osteoporosis risk. METHODS: We searched eligible studies about ApaI polymorphism and osteoporosis through the PubMed, Embase, CNKI and Wanfang databases; case-control studies containing available genotype frequencies of A/a were chosen. We used the odds ratio with 95% confidence interval to assess the strength of this association. Sensitivity analysis and publication bias assessment were performed. Trial sequential analysis (TSA) was performed to evaluate a sufficient sample. RESULTS: Twenty-two studies assessed the relationship between ApaI polymorphism and the risk of osteoporosis in postmenopausal women. The comprehensive analyses showed no significant association for ApaI polymorphism with postmenopausal osteoporosis in the overall population, equally valid for Asian and Caucasian subgroups with any genetic model. TSA still indicated the results were robust. CONCLUSION: The present meta-analysis suggests that the VDR ApaI genotype may not affect the risk of postmenopausal osteoporosis in Asians and Caucasians.

[Progress of researches on roles of dendritic cells in immune tolerance caused by Echinococcus infections].

Dendritic cells (DCs), a type of antigen-presenting cells (APC), are recognized as an important regulator of immune response and immune tolerance, and play a critical role in the host innate immunity and adaptive immunity. Previous studies have shown that the long-term parasization of Echinococcus in the host is strongly associated with the host immune tolerance induced by DCs. This review summarizes the research progress of the role of DCs in host immune tolerance caused Echinococcus infection, aiming to provide the theoretical basis and insights into the management and immunotherapy of Echinococcus infections.

3,4-Methylenedioxymethamphetamine causes retinal damage in C57BL/6J mice.

3,4-Methylenedioxymethamphetamine (MDMA) is a powerfully addictive psychostimulant with pronounced effects on the central nervous system, but the precise mechanism of MDMA-induced toxicity remains unclear, specifically on the retina. This study was performed to investigate the effects of MDMA treatment on the retina and explore the underlying mechanism. C57BL/6J mice were randomly divided into control and MDMA groups. Mice were treated with MDMA at progressively increasing doses (1-6 mg/kg) intraperitoneally 4 times per day. Electroretinography was used to test the retinal function. Pathological changes of the retina were examined by toluidine blue staining and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling assay. Enzyme-linked immunosorbent assays were used to measure the levels of cytokines in the retina. Real-time polymerase chain reaction and Western blot were used to measure gene and protein expression in the retina, respectively. Our study showed that MDMA treatment impaired retinal function and decreased retinal thickness. MDMA treatment also increased transforming growth factor ß as well as inflammatory factors in the retina. Moreover, MDMA treatment increased protein expression of matrix metalloproteinases (MMPs) and decreased tight junction protein expression in the retina. Our study indicated that treatment of MDMA caused retinal damage in C57BL/6J mice, associated with an increase of MMPs and a decrease of tight junction proteins.

A novel near-infrared nanomaterial with high quantum efficiency and its applications in real time in vivo imaging.

Fluorescence imaging signal is severely limited by the quantum efficiency and emission wavelength. To overcome these challenges, novel NIR-emitting K5NdLi2F10 nanoparticles under NIR excitation was introduced as fluorescence imaging probe for the first time. The photostability of K5NdLi2F10 nanoparticles in the water, phosphate buffer saline, fetal bovine serum and living mice was investigated. The fluorescence signal was detected with depths of 3.5 and 2.0 cm in phantom and pork tissue, respectively. Fluorescence spectrum with a significant signal-to-background ratio of 10:1 was captured in living mice. Moreover, clear NIR images were virtualized for the living mice after intravenous injection. The imaging ability of nanoparticles in tumor-beard mice were evaluated, the enrichment of K5NdLi2F10 nanoparticles in tumor site due to the enhanced permeability and retention effect was confirmed. The systematic studies of toxicity, bio-distribution and in-vivo dynamic imaging suggest that these materials give high biocompatibility and low toxicity. These NIR-emitting nanoparticles with high quantum efficiency, high penetration and low toxicity might facilitate tumor identification in deep tissues more sensitively.

A retrospective analysis of ovarian stimulation with letrozole in women undergoing artificial insemination by donor.

The aim of this retrospective study was to determine the clinical pregnancy rate in women undergoing letrozole ovarian stimulation and artificial insemination by donor (AID). Between 2012 and 2015, 130 natural cycles, 939 letrozole cycles and 130 letrozole plus gonadotrophin cycles were conducted. Letrozole cycles were divided into three groups according to LH concentration on the day of HCG administration (LH <10 mIU/ml and follicle size ≥18 cm; LH ≤10 to <20 mIU/ml; and LH ≥20 mIU/ml). Pregnancy rates were 17.3%, 22.4% and 26.8%, respectively (P = 0.012). In women given 10 mIU/ml LH or more, logistic regression identified oestradiol (OR 1.002, 95% CI, 1.000 to 1.004, P = 0.029) and leading follicle size (OR 0.861, 95% CI, 0.772 to 0.960, P = 0.007) as significant predictive factors of pregnancy rate; the higher the oestradiol and the smaller the follicles, the better the pregnancy rate. The pregnancy rate was significantly higher in the letrozole plus gonadotrophin group than the letrozole group (P = 0.04). Better pregnancy rates can be achieved if LH surge occurs before HCG administration, especially with higher oestradiol and lower follicle size; treatment with letrozole plus gonadotrophin was significantly more effective than letrozole alone in AID.

MicroRNA-32 functions as a tumor suppressor and directly targets EZH2 in uveal melanoma.

MicroRNA-32 (miR-32) has been shown to be dysregulated in some human malignancies and this has been found to be correlated with tumor progression. However, its role in uveal melanoma formation and progression remains largely unknown. Thus, the aim of this study was to explore the expression and function of miR-32 in human uveal melanoma. Using quantitative reverse transcription-polymerase chain reaction, we detected miR-32 expression in uveal melanoma tumor tissues and cell lines. The effects of miR-32 on the biological behavior of uveal melanoma cells were also investigated. Finally, the potential regulatory function of miR-32 on EZH2 expression was confirmed. miR-32 expression levels were significantly downregulated in uveal melanoma samples and cell lines (both P < 0.01). Ectopic expression of miR-32 could inhibit uveal melanoma cell proliferation, migration, and invasion, and promote cell apoptosis in vitro. Further, EZH2 was confirmed as a direct target of miR-32 by using the luciferase reporter assay. These findings indicate that miR-32 may function as a novel tumor suppressor in uveal melanoma and could be a potential therapeutic target for this disease.

Correlations of leukemia inhibitory factor and macrophage migration inhibitory factor with endometrial carcinoma.

OBJECTIVE: To investigate the correlations of leukemia inhibitory factor (LIF) and macrophage migration inhibitory factor (MIF) with endometrial carcinoma. MATERIALS AND METHODS: The study included 113 endometrial specimens from the Fourth Affiliated Hospital of Harbin Medical University, collected from May 2006 to October 2008, classified into normal endometrium, simple hyperplasia, complex hyperplasia, atypical hyperplasia, and endometrial carcinoma. The LIF and MIF expression of all 113 specimens was detected with immunohistochemistrical (IHC) method. RESULTS: The MIF expression in hyperplastic endometrium and endometrial carcinoma increased significantly as compared with that in normal endometrium (p < 0.05 and p < 0.001, respectively), and its expression in endometrial carcinoma was also remarkably higher than that in hyperplastic endometrium (p < 0.001). The expressions of LIF in atypical hyperplasia and endometrial carcinoma were also significantly higher than that in the normal endometrium (p < 0.05), but it is not obviously higher in simple hyperplasia and complex hyperplasia than in the normal endometrium (p > 0.05). Furthermore, the expression of LIF showed no statistical difference between hyperplastic endometrium and endometrial carcinoma. CONCLUSION: It could be speculated that MIF may be correlated with the occurrence of endometrial carcinoma. However, whether LIF also has a correlation with the occurrence of endometrial carcinoma still cannot be presumed.

Selective estrogen receptor modulators promising for cardiac syndrome X.

Cardiac syndrome X (CSX) is defined as a typical anginal-like chest pain with a transient ischemic electrocardiogram, but without abnormal coronary angiography. It is usually accepted that endothelial dysfunction, inflammation, oxidative stress and estrogen deficiency are the main reasons of CSX. There are some methods to treat CSX including statins, b blocker, angiotensin converting enzyme inhibitors, nitrates, estrogen, and so on. The estrogen replacement therapy (ERT), in particular, has been reported by many researchers to significantly reduce the frequency of chest pain after administration of estrogen, which has been explained as estrogen acting on its receptor to improve the endothelial function. However, it has been suggested that ERT must not be used for coronary heart disease due to its adverse effects. However, some selective estrogen receptor modulators (SERMs) can inhibit inflammatory response as well as oxidative stress, and improve the endothelial function, to reduce the occurrence of chest pain. Here, we hypothesize that SERMs may be the beneficial selection for patients with CSX.

Inhibiting extracellular matrix metalloproteinase inducer maybe beneficial for diminishing the atherosclerotic plaque instability.

Atherosclerotic plaque rupture and local thrombosis activation in the artery cause acute serious incidents such as acute coronary syndrome and stroke. The exact mechanism of plaque rupture remains unclear but excessive degradation of the extracellular matrix scaffold by matrix-degrading metalloproteinases (MMPs) has been implicated as one of the major molecular mechanisms in this process. Convincing evidence is available to prove that extracellular matrix metalloproteinase inducer (EMMPRIN) induces MMP expression and is involved in the inflammatory responses in the artery wall. The inflammation and MMPs have been shown to play a critical role for atherosclerotic lesion development and progression. More recent data showed that increased EMMPRIN expression was associated with vulnerable atherosclerotic lesions. Therefore, we speculate that EMMPRIN may be pivotal for atherosclerotic plaque instability, and hence inhibition of EMMPRIN expression could be a promising approach for the prevention or treatment of atheroma instability.

Glucometabolic state of in-hospital patients with primary hypertension in sub-population of partial regions in China.

Abnormal glucose metabolism (AGM) is common but underestimated in patients with coronary heart disease (CHD). Here, we reported 898 in-hospital patients with primary hypertension (PH) at the university hospitals in developed regions of China. Oral glucose tolerance test (OGTT) was performed in those without known type-2 diabetes mellitus (2-DM). A total of 158 patients had known 2-DM and 32 were newly diagnosed as 2-DM by fasting blood glucose (FBG). OGTT revealed that 83 had 2-DM and 296 had impaired glucose tolerance (IGT). The proportion of 2-DM and AGM increased from 21.2 to 30.4% and from 57.5 to 68.7% upon OGTT. Prevalence of AGM and 2-DM increased with the increase of age, and incidence of AGM and 2-DM was significant higher in patients with risk factors (including CHD, overweight, hyperlipidaemia, proteinuria) than those without risk factors mentioned above. Glucose was not sufficiently controlled in 55.1% of the patients with 2-DM upon treatment, well controlled in 35.4% and not controlled in 9.5%. So AGM is also prevalent in PH patients especially the elders and those with risk factors, which was underestimated in most cases. Moreover, much lower awareness, treatment and control of 2-DM occurred in some regions of China, thus strengthening health education for patients and heightening consciousness of doctor are very important and eminent. Except for FBG, more attention should be paid to postprandial blood glucose ignored before, and OGTT should be a routine procedure in PH patients, especially in older patients and those with the factors mentioned above.

Estrogen action and male fertility: roles of the sodium/hydrogen exchanger-3 and fluid reabsorption in reproductive tract function.

Estrogen receptor alpha (ER alpha) is essential for male fertility. Its activity is responsible for maintaining epithelial cytoarchitecture in efferent ductules and the reabsorption of fluid for concentrating sperm in the head of the epididymis. These discoveries and others have helped to establish estrogen's bisexual role in reproductive importance. Reported here is the molecular mechanism to explain estrogen's role in fluid reabsorption in the male reproductive tract. It is shown that estrogen regulates expression of the Na(+)/H(+) exchanger-3 (NHE3) and the rate of (22)Na(+) transport, sensitive to an NHE3 inhibitor. Immunohistochemical staining for NHE3, carbonic anhydrase II (CAII), and aquaporin-I (AQP1) was decreased in ER alpha knockout (alpha ERKO) efferent ductules. Targeted gene-deficient mice were compared with alpha ERKO, and the NHE3 knockout and CAII-deficient mice showed alpha ERKO-like fluid accumulation, but only the NHE3 knockout and alpha ERKO mice were infertile. Northern blot analysis showed decreases in mRNA for NHE3 in alpha ERKO and antiestrogen-treated mice. The changes in AQP1 and CAII in alpha ERKO seemed to be secondary because of the disruption of apical cytoarchitecture. Ductal epithelial ultrastructure was abnormal only in alpha ERKO mice. Thus, in the male, estrogen regulates one of the most important epithelial ion transporters and maintains epithelial morphological differentiation in efferent ductules of the male, independent of its regulation of Na(+) transport. Finally, these data raise the possibility of targeting ER alpha in developing a contraceptive for the male.

Human estrogen receptor beta-specific monoclonal antibodies: characterization and use in studies of estrogen receptor beta protein expression in reproductive tissues.

Investigation of the role of the second, more recently described estrogen receptor, denoted ERbeta, will be critical in understanding the molecular mechanisms underlying tissue-specific gene regulation by estrogens. Expression of ERbeta in a variety of tissues has been examined predominantly at the mRNA level, and there is little information regarding the cellular localization and size of the endogenous ERbeta protein, due, in part, to the limited availability of human ERbeta-specific antibodies. Thus, our aim was to generate specific antibodies to human ERbeta and use them to determine the tissue-specific distribution and size(s) of the ERbeta protein. To this end, we have cloned three different hybridoma cell lines that produce monoclonal antibodies specific for the hormone-binding domain of human ERbeta. The antibodies, made in mice against human ERbeta amino acids 256-505 (hormone binding domain lacking the F domain), are designated CFK-E12 (E12), CMK-A9 (A9) and CWK-F12 (F12) and were determined to be the IgG gamma1 isotype for E12, and IgG gamma2b for A9 and F12. All three monoclonal antibodies could be used to detect in vitro translated, baculovirus expressed, and cell transfected and expressed ERbeta protein by Western blot analyses, and all failed to detect ERalpha. A9 and F12 were able to immunoprecipitate efficiently the native form of ERbeta protein in the presence and absence of estradiol. Epitope mapping studies indicate that the E12 and F12 antibodies recognize overlapping peptide sequences in the N-terminal region of the hormone-binding domain, a region that is highly conserved among species. Immunocytochemical studies with these antibodies reveal nuclear-specific localization of the ERbeta protein in granulosa cells of the rat ovary. Nuclear ERbeta is also specifically localized in epithelial and some stromal cells of mouse and rat epididymis. Western blot analysis with protein extracts from ovarian granulosa cells of human, rat, mouse, and pig showed a ca. 52 kDa and an additional ca. 62-64 kDa band in these species. These results indicate the presence of two predominant molecular size forms of the ERbeta protein in ovarian granulosa cells and demonstrate the utility of these antibodies for detection of ERbeta in the human and in several other mammalian species.

Morphological analysis of endocytosis in efferent ductules of estrogen receptor-alpha knockout male mouse.

Lack of estrogen receptor (ER) results in fluid accumulation and dilation of the efferent ductules, suggesting that the role of estrogen and ER in the male reproductive tract is related to fluid reabsorption in the ductules. In the present study, endocytosis of the nonciliated cells of the efferent ductules was compared morphologically between wild type (WT) and estrogen receptor-alpha knockout (alpha ERKO) male mice. The epithelial cells lining the WT efferent ductules were tall columnar in shape, whereas those of the alpha ERKO were low columnar. Immunocytochemically, the nonciliated cells of both genotypes showed positive reactions of sulfated glycoprotein-2, but the reaction products were reduced in amount in the alpha ERKO. Electron microscopy revealed that the nonciliated cells of the WT had numerous organelles for endocytosis such as coated pits and vesicles, tubules, endosomes, multivesicular bodies and lysosomes in the apical cytoplasm. These organelles were less developed in the nonciliated cells of the alpha ERKO. Morphometric analysis indicated that there was a significant reduction in area of endocytotic apparatus in the nonciliated cells of the alpha ERKO compared with that of the WT. A tracer study using gold particles demonstrated that the nonciliated cells of both WT and alpha ERKO efferent ductules were capable of taking up luminal contents. These results suggest that reabsorption of the luminal contents via endocytosis takes place in the efferent ductules but is greatly reduced in amount in the absence of ER alpha.

Kupffer cell-initiated remote hepatic injury following bilateral hindlimb ischemia is complement dependent.

Intravital fluorescence microscopy was applied to the livers of male Wistar rats to test the hypothesis that complement mobilization stimulates Kupffer cells and subsequently initiates hepatic injury after hindlimb ischemia/reperfusion (I/R). Following 3 h of limb reperfusion, hepatocellular viability (serum levels of alanine transaminase and cell death via propidium iodide labeling) decreased significantly from levels in sham-operated animals. Inhibition of complement mobilization with soluble complement receptor type 1 (20 mg/kg body wt) and interruption of Kupffer cell function with GdCl(3) (1 mg/100g body wt) resulted in significant hepatocellular protection. Although the effects of hindlimb I/R on hepatic microvascular perfusion were manifest as increased heterogeneity, both complement inhibition and suppression of Kupffer cell function resulted in marked improvements. No additional hepatocellular protection and microvascular improvements were provided by combining the interventions. Furthermore, inhibition of complement mobilization significantly depressed Kupffer cell phagocytosis by 42% following limb reperfusion. These results suggest that the stimulation of Kupffer cells via complement mobilization is necessary but is not the only factor contributing to the early pathogenesis of hepatic injury following hindlimb I/R.

Estrogens and epididymal function.

Estrogen is synthesized in the male reproductive system and is found in high concentrations in rete testis and seminal fluids. This luminal estrogen targets estrogen receptors (ER) along the male reproductive tract, and in particular the efferent ductules, where ERalpha is abundant. However, both ERalpha and ERbeta are found in various regions of the male reproductive tract. The transgenic ER knockout mice (alphaERKO and betaERKO) have been used to help define the role of ER in the male. In the alphaERKO animal model, the efferent ductules are dramatically altered, forming an epithelium in which fluid reabsorption is inhibited and epithelial cells have greatly reduced numbers of lysosomes and organelles associated with endocytosis. The betaERKO male reproductive tract appears normal. Because these animals are transgenic and lack ER throughout development, we developed animal models using pure antiestrogen ICI 182,780 treatments in adult males. The data show that ERalpha participates in the regulation of the apical cytoplasm of non-ciliated cells of the efferent ductules, narrow cells of initial segment epididymis and clear cells in the remaining segments of the epididymis. There appears to be no effect on vas deferens. The inhibition of ERalpha function in the male leads to decreases in sperm concentrations and eventually to infertility. The current literature leaves the mechanisms of estrogen action in the male reproductive tract unsettled and raises the question of androgen's contribution to the regulation of fluid transport, especially in the efferent ductules.

Influence of restricted food intake on estrous cycles and pseudopregnancies in dogs.

OBJECTIVE: To examine effects of restricted food intake on estrous cycle frequency, interestrus interval, and pseudopregnancy prevalence in dogs. ANIMALS: 28 female Labrador Retrievers. PROCEDURE: Dogs were paired by body weight when they were 6 weeks old and fed so that the limit-fed pair-mate received 75% of the amount of food offered to its maintenance-fed counterpart. Estrous cycle, interestrus interval, and pseudopregnancy data were recorded. RESULTS: Mean annual frequency of estrous cycles and duration of interestrus intervals did not differ between feeding groups. Prevalence of clinically evident pseudopregnancy was significantly greater among females that were maintenance fed, although results of endocrinologic testing did not identify a mechanism for this observation. CONCLUSIONS AND CLINICAL RELEVANCE: Pseudopregnancy in dogs can be influenced by physiologic factors related to nutrition. Clinicians should consider a variety of physiologic and environmental factors when evaluating reproductive function in dogs.

Three steroidal alkaloids from Buxus microphylla.

Three new steroidal alkaloids have been isolated from the leaves and stems of Buxus microphylla Sieb et Zucc. They are buxmicrophylline B, C and D. Their structures were elucidated by extensive analysis of the spectral data.

Within- and between-examiner repeatability of distraction indices of the hip joints in dogs.

OBJECTIVE: To evaluate in vivo repeatability of the distraction index method of evaluating hip joint laxity in dogs. ANIMALS: 31 two-year-old Labrador Retrievers. PROCEDURE: Each dog was anesthetized and radiographically evaluated for hip joint laxity 4 times: twice by an experienced examiner and twice by an examiner who had no previous knowledge of or training in the technique prior to the first day of testing. Distraction indices (DI) were determined from the radiographs and intraclass correlation coefficients were calculated to evaluate the repeatability of DI measurements between and within examiners. RESULTS: Intraclass correlation coefficients were high (range, 0.85 to 0.94). Lower limits of the 95% confidence intervals for the intraclass correlation coefficients ranged from 0.75 to 0.89. CONCLUSIONS: Between- and within-examiner repeatabilities of DI measurements were high, suggesting that the technique is clinically reliable. CLINICAL RELEVANCE: Distraction index is a reliable measure of hip joint laxity and a good predictor of the risk of development of degenerative joint disease associated with hip dysplasia in dogs. Establishment of high repeatability of DI measurements suggests that the stress-radiographic method may be used by multiple examiners with the expectation of comparable and consistent results.