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.

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.

[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.

Localization of a gene for expression of mouse mammary tumor virus antigens in the GR/Mtv-2- mouse strain.

The GR/Mtv-2- mouse strain is congenic to the GR strain but lacks the Mtv-2 gene for high amounts of mouse mammary tumor virus (MMTV) virion particles in the milk and early mammary tumors. With a sensitive competition radioimmunoassay for individual viral proteins of MMTV, substantial amounts of the gag proteins p27 and p10 could still be detected in extracts of the mammary glands of GR/Mtv-2- mice, but essentially no viral envelope antigens. The genetic transmission of the MMTV gag expression in the GR/Mtv-2- strain was investigated. In a cross with the virus-negative BALB/c strain, the MMTV p27 expression behaved as a dominant feature. Double backcross analysis proved that the p27 expression was governed by a single gene located on chromosome 11, cloe to the Es-3 locus. The gene was thereby not allelic to any of the previously described MMTV induction genes, Mtv-1 and Mtv-2, and is therefore called Mtv-3. It is concluded that the total MMTV expression in the GR strain is under control of two separate loci, Mtv-2 on chromosome 18, inducing high levels of complete virus particles and also early mammary tumors; and Mtv-3 on chromosome 11, coding for partial MMTV expression.

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.

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.

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.

Genetic analysis of mammary tumor induction and expression of mammary tumor virus antigen in hormone-treated ovariectomized GR mice.

Early stages of mammary tumors (EMT) were induced with a combined treatment of progesterone (P) and estrone (E) in ovariectomized adult GRS/A (GR) mice, a strain of European origin and with a high incidence of mammary cancer. The mammary tumors were comparable to the pregnancy-dependent tumors of breeding females of this strain. The hormone treatment did not lead to EMT in a variety of other strains and only occasionally in the RIII an C3H strains. Treatment with P or E alone di not lead to EMT in GR mice, but treatment with the steroid compound 17 alpha-ethynyl-19-nortestosterone (ANT) did mimic the combined effe(t of P and E. Since EMT could be induced by ANT in all ovariectomized adult GR mice within 3 weeks, this tumor-induction method was suitable for analysis of the gene responsible for palpable, pregnancy-dependent mammary tumors of this strain. Another argument for the usefulness of this test for genetic analysis was the fact that, though mouse mammary tumor virus (MuMTV) of the GR strain was introduced into BALB/c and tmas mice by foster-nursing, ANT treatment did not lead to EMT. First and second backcross analyses showed that one gene was responsible for EMT induction. There was a strong (orrelation between the presence of EMT and MuMTV antigens in the mammary glands and milk of several first backcross populations between GR and other strains such as C57BL, BALB/c, DBAf, and C3Hf. This suggested that the expression of MuMTV antigens was also controlled by the EMT gene. Two types of resistance phenomena were observed. Neither type could prevent EMT after hormone treatment; however, they could delay EMT development. One resistance factor for EMT induction was noticeable and dominant in reciprocal hybrids of the GR and DBAf strains, whereas another resistance factor was detected in the backcross population only [i.e., in the C57BL X (C57BL X Gr) ba(kcross] and not in hybrids; therefore, this factor was recessive. Until now, linkage experiments with 18 markers to locate the gene for EMT induction in the map of the mouse were unsuccessful.

Mouse strain (STS/A) resistant to mammary tumor induction by hypophysial isografts.

Implantation of hypophysial isografts does not lead to induction of mammary tumors in all strains of mice lacking the exogenous murine mammary tumor virus. While O20, C3Hf, and BALB/c females are highly susceptible and C57BL and TSI females are of intermediate susceptibility, the STS females appear to be nearly totally resistant. The resistance of the STS strain is not due to failure of prolactin production by the hypophysial isografts and may therefore be due to a genetically controlled mechanism at target cell level. Neither resistance, i.e., low incidence of mammary tumors (2% in STS), nor susceptibility, i.e., high incidence at low age (93% at 349 days in C3Hf; 83% at 360 days in BALB/c), is dominant. F1 hybrids of strain STS and the two strains C3Hf and BALB/c show high incidences (STS X C3Hf F1, 90%; STS X BALB/c F1, 60%), but the age at which tumors appear (476 and 604 days, respectively) is much higher, suggesting that more than one gene is involved in this type of hormonal carcinogenesis of the mammary gland in mice.

Membrane glycoprotein changes in primary mammary tumors associated with autonomous growth.

Primary and transplanted mammary tumors of the GR mouse were explanted in tissue culture and grown in the presence of radioactive fucose. Labelled membrane glycopeptides were isolated and compared by cochromatography with differentially labelled glycopeptides from normal mammary gland tissue. Differences with controls in the glycopeptide elution profiles were observed in autonomous, hormone-independent tumors but were absent in histologically similar tumors which required a continuous hormonal stimulus for growth. The results suggest that alterations in membrane glycopeptides are associated with the capacity of autonomous, hormone-independent growth of murine adenocarcinoma cells.

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.

Two triterpenoid saponins from Pterocephalus bretschneidri.

Two new triterpenoid saponins, named bretschnosides A and B, were isolated from the roots of Pterocephalus bretschneidri. On the basis of chemical degradation and spectroscopic evidence, the structures of bretschnosides A and B were shown to be 3-O-alpha-L-rhamnopyranosyl(1-->3)- beta-D-xylopyranosyl(1-->3)-alpha-L-rhamnopyranosyl(1-->2)-beta-D- xylopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl(1-->6)-beta-D-glucopyranoside 4 and 3-O-alpha-D-glucopyranosyl(1-->3)-beta-D-xylopyranosyl(1-->3)-alpha-L- rhamnopyranosyl(1-->2)-beta-D-xylopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl(1-->6)-beta-D-glucopyranoside 6, respectively.

Triterpenoid saponins from Pterocephalus hookeri.

Four new triterpenoid saponins, named hookerosides A-D, were isolated from Pterocephalus hookeri. Based on chemical and spectral evidence, their structures were determined as 3-O-beta-D-glucopyranosyl(1-->4)-beta-D-xylopyranosyl(1-->3)-alpha-L- Rhamnopyranosyl(1-->2)-beta-D-xylopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl(1-->6)-beta-D-glucopyranoside, 3-O-beta-D-xylopyranosyl(1-->4)-beta-D-glucopyranosyl(1-->4)-beta-D- xylopyranosyl(1-->3)-alpha-L-rhamnopyranosyl(1-->2)-beta-D-xylopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl(1-->6)-beta-D-glucopyranoside, 3-O-alpha-L-rhamnopyranosyl(1-->2)-beta-D-xylopyranosyl(1-->4)-beta-D- glucopyranosyl(1-->4)-beta-D-xylopyranosyl(1-->3)-alpha-L-rhamnopyranosy l(1-->2 ) beta-D-xylopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl(1-->6)-beta-glucopyranoside and oleanolic acid 3-O-alpha-L-rhamnopyranosyl(1-->2)-beta-D- xylopyranosyl(1-->4)-beta-D-glucopyranosyl(1-->4)-beta-D-xylopyranosyl(1 -->3)- alpha-L-rhamnopyranosyl(1-->2)-beta-D-xylopyranoside, respectively.

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.

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.