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Testes control the development of male reproductive system. The testicular interstitial Leydig cells (Leydig cells) synthesize testosterone for promoting spermatogenesis and secondary sexual characteristics. Type A platelet-derived growth factor (PDGF-AA) is one of the most important growth factors in regulating Leydig cell growth and function. Knockout of PDGF-AA or its congenital receptor PDGFR-α leads to poor testicular development caused by reducing Leydig cell numbers, supporting PDGF-AA/PDGFR-α signaling regulates Leydig cell development. Primary cilium is a cellular antenna that functions as an integrative platform to transduce extracellular signaling for proper development and differentiation. Several receptors including PDGFR-α are observed on primary cilia for initiating signaling cascades in distinct cell types. Here we showed that PDGF-AA/PDGFR-α signaling promoted Leydig cells growth, migration, and invasion via primary cilia. Upon PDGF-AA treatment, AKT and ERK signaling were activated to regulate these cellular events. Interestingly, active AKT and ERK were detected around the base of primary cilia. Depletion of ciliary genes (IFT88 and CEP164) alleviated PDGF-AA-activated AKT and ERK, thus reducing Leydig cell growth, migration, and invasion. Thus, our study not only reveals the function of PDGF-AA/PDGFR-α signaling in maintaining testicular physiology but also uncovers the role of primary cilium and downstream signaling in regulating Leydig cell development.

The connecting pieces of the sperm neck link the flagellum and the sperm head, and they are important for initiating flagellar beating. The connecting pieces are important building blocks for the sperm neck; however, the mechanism of connecting piece assembly is poorly understood. In the present study, we explored the role of septins in sperm motility and found that Sept12D197N knock-in (KI) mice produce acephalic and immotile spermatozoa. Electron microscopy analysis showed defective connecting pieces in sperm from KI mice, indicating that SEPT12 is required for the establishment of connecting pieces. We also found that SEPT12 formed a complex with SEPT1, SEPT2, SEPT10 and SEPT11 at the sperm neck and that the D197N mutation disrupted the complex, suggesting that the SEPT12 complex is involved in the assembly of connecting pieces. Additionally, we found that SEPT12 interacted and colocalized with γ-tubulin in elongating spermatids, implying that SEPT12 and pericentriolar materials jointly contribute to the formation of connecting pieces. Collectively, our findings suggest that SEPT12 is required for the formation of striated columns, and the capitulum and for maintaining the stability of the sperm head–tail junction.

The aim of this study is to investigate the frequency and distribution of human HLA sharing and maternal HLA allele expression in couples with recurrent pregnancy loss in Taiwan.We retrospectively reviewed couples experienced two or more pregnancy loss before 20th weeks of gestation from March 2014 to November 2020 having HLA determination. Fertile individuals with one or more live-birth offspring receiving HLA allele determination during the same period were included as the control group. The distribution and frequency of HLA sharing were analyzed and presented by descriptive statistics. Fisher Exact Test were used to analyze specific maternal and paternal HLA allele comparing individuals with RPL to fertile group. P-value0.05 was thought to be statistically significant.72 couples were enrolled from March 2014 to November 2020. Regarding the distribution of HLA sharing, HLA sharing between females and their male partners less and equal to 2 pairs were found in 40.3% of the couples. HLA sharing greater and equal to 3 pairs are found in 59.8% couples. HLA sharing was most frequently found in alleles HLA-A02, A11, DQ07, C07 and B60 in descending order. There was a significant lower expression of HLA-B13 in women with RPL compared to women who had successful pregnancy (p = 0.0234). Compared infertile men with fertile men cohort, the frequency of HLA-DR04 (p = 0.0438, OR 2.444, 95% CI 1.0251-5.8287), HLA-DR12 (p = 0.001, OR 30.85, 95% CI 4.0296-236.19) and HLA-15 (p = 0.0357, OR 9.354, 95% CI 1.1610-75.37) were found to be significantly higher in men with RPL. On the contrary, HLA-DR07 (p = 0.0085, OR 0.124, 95% CI 0.0264-0.587) and HLA-DR10 (p = 0.0395, OR 0.048, 95% CI 0.0027-0.8641) were found to be significantly lower in men with RPL.We found a tendency to recurrent pregnancy loss in couples with more than 2 pairs of HLA sharing. The similarity of HLA sharing, the expression of maternal HLA-B13 allele and paternal HLA-DR alleles in Taiwanese couples might play a role in recurrent pregnancy loss.

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Sperm growth and maturation are correlated with the expression levels of Leucine-rich repeat and WD repeat-containing protein 1 (LRWD1), a widely expressed protein in the human testicles. The decrease in LRWD1 cellular level was linked to the reduction in cell growth and mitosis and the rise in cell microtubule atrophy rates. Since DNA methylation has a major regulatory role in gene expression, this study aimed at exploring the effect of the modulation of DNA methylation on LRWD1 expression levels.The results revealed the presence of a CpG island up of 298 bps (- 253 ~ + 45) upon LRWD1 promoter in NT2/D1 cells. The hypermethylation of the LRWD1 promoter was linked to a reduction in the transcription activity in NT2/D1 cells, as indicated by luciferase reporter assay. The methylation activator, floxuridine, confirmed the decrease in the LRWD1 promoter transcriptional activity. On the other hand, 5-Aza-2'-deoxycytidine (5-Aza-dc, methylation inhibitor), significantly augmented LRWD1 promoter activity and the expression levels of mRNA and proteins. Furthermore, DNA methylation status of LRWD1 promoter in human sperm genomic DNA samples was analyzed. The results indicated that methylation of LRWD1 promoter was correlated to sperm activity.Thus, the regulation of LRWD1 expression is correlated with the methylation status of LRWD1 promoter, which played a significant role in the modulation of spermatogenesis, sperm motility, and vitality. Based on these results, the methylation status of LRWD1 promoter may serve as a novel molecular diagnostic marker or a therapeutic target in males' infertility.RéSUMé: CONTEXTE: La croissance et la maturation des spermatozoïdes sont corrélées avec les niveaux d’expression de la protéine 1 riche en répétitions Leucine et contenant des répétitions WD (LRWD1), une protéine largement exprimée dans les testicules humains. La diminution du niveau cellulaire en LRWD1 a été liée à une réduction de la croissance et des mitoses cellulaires, et à une augmentation des taux d’atrophie des microtubules cellulaires. Puisque la méthylation de l’ADN joue un rôle régulateur majeur dans l’expression des gènes, cette étude visait à explorer l’effet de la modulation de la méthylation de l’ADN sur les niveaux d’expression de LRWD1. RéSULTATS: Les résultats ont révélé la présence d’un îlot CpP de 298 pbs (-253~+45) sur le promoteur de LRWD1dans les cellules NT2/D1. L’hyperméthylation du promoteur de LRWD1 était liée à une réduction de l’activité de transcription dans les cellules NT2/D1, comme indiqué par l’analyse de l’expression d’un gène rapporteur codant pour la luciférase. L’activateur de méthylation, la floxuridine, a confirmé la diminution de l’activité transcriptionnelle du promoteur de LRWD1. D’autre part, la 5-Aza-2'-déoxycytidine (5-Aza-dc, inhibiteur de méthylation), a significativement augmenté l’activité du promoteur de LRWD1 et les niveaux d’expression de l’ARNm et des protéines. En outre, le statut de méthylation de l’ADN du promoteur de LRWD1 dans les échantillons d’ADN génomique de sperme humain a été analysé. Les résultats ont indiqué que la méthylation du promoteur de LRWD1 était corrélée à l’activité des spermatozoïdes. CONCLUSIONS: Ainsi, la régulation de l’expression LRWD1 est corrélée avec le statut de méthylation du promoteur de LRWD1, qui a joué un rôle important dans la modulation de la spermatogenèse, de la mobilité et de la vitalité des spermatozoïdes. Sur la base de ces résultats, le statut de méthylation du promoteur de LRWD1 peut servir de nouveau marqueur diagnostic moléculaire ou de cible thérapeutique dans l’infertilité masculine.

Irisin has gained attention because of its potential applications in the treatment of metabolic diseases. Accumulating evidence indicates that irisin attenuates obesity via the browning of white adipose tissue; however, the underlying mechanisms are unclear. Here, we evaluated the effects of irisin on adipocyte browning and the underlying mechanisms. The western blotting and immunofluorescence analyses demonstrated that irisin significantly induced the up-regulation of brown fat-specific proteins (PGC1α, PRDM16, and UCP-1) and HO-1 in 3T3-L1 adipocytes. Moreover, irisin significantly increased the levels of cytosolic p62 and nuclear Nrf2. These effects of irisin in the adipocytes were attenuated by treatment with SnPP or p62 siRNA. In addition, the browning effect of irisin was observed in BAT-WT-1 cells. These findings suggest that irisin induced browning effect via the p62/Nrf2/HO-1 signalling pathway and that it may be a potential candidate for preventing or treating obesity.

This study aimed to determine the antiobesity effects of raspberry ketone (RK), one of the major aromatic compounds contained in raspberry, and its underlying mechanisms. During adipogenesis of 3T3-L1 cells, RK (300 μM) significantly reduced lipid accumulation and downregulated the expression of CCAAT/enhancer-binding protein α (C/EBPα), peroxisome proliferation-activated receptor γ (PPARγ), fatty acid-binding protein 4 (FABP4), and fatty acid synthase (FAS). RK also reduced the expression of light chain 3B (LC3B), autophagy-related protein 12 (Atg12), sirtuin 1 (SIRT1), and phosphorylated-tuberous sclerosis complex 2 (TSC2), whereas it increased the level of p62 and phosphorylated-mammalian target of rapamycin (mTOR). Daily administration of RK decreased the body weight (ovariectomy [Ovx] + RK, 352.6 ± 5 vs Ovx, 386 ± 5.8 g; P < 0.05), fat mass (Ovx + RK, 3.2 ± 0.05 vs Ovx, 5.0 ± 0.4 g; P < 0.05), and fat cell size (Ovx + RK, 6.4 ± 0.6 vs Ovx, 11.1 ± 0.7 × 103 μm2; P < 0.05) in Ovx-induced obesity in rat...

M1 macrophage differentiation plays a crucial role in enhanced inflammation during pregnancy, which may lead to pregnancy complications. Therefore, modulation of macrophage differentiation toward the M2 phenotype is desirable to ensure a successful pregnancy. Medroxyprogesterone acetate (MPA) is a potent progestin with an anti-inflammatory property, but its effect on macrophage differentiation is unknown. This study aimed to examine whether MPA can induce an M2 macrophage differentiation by using the human monocytes cell line THP-1 or primary monocytes. THP-1 cells were primed with phorbol-12-myristate-13 acetate (PMA) to initiate macrophage differentiation. By incubating with MPA, the cells (denoted as MPA-pTHP-1) underwent M2 macrophage differentiation with downregulations of CD11c, IL-1β and TNF-α, and upregulations of CD163 and IL-10; while cells incubated with progesterone (P4) did not show the M2 phenotype. Primary monocytes treated with MPA also had the same M2 phenotype. Moreover, M1 macrophages derived from IFN-γ/LPS-treated THP-1 cells, which had high levels of IL-1b and iNOS, and low levels of IL-10 and IDO, were reversed to the M2 phenotype by the MPA treatment. We also found that the MPA-pTHP-1 promoted the decidualization of endometrial stromal cells and the invasion of trophoblast cells. To mimic conditions of exposure to various pathogens, MPA-pTHP-1 cells were stimulated by different types of TLR ligands. We found they produced lower levels of IL-1β and TNF-α, as well as a higher level of IL-10, compared to untreated cells. Finally, we found the level of phosphorylated ERK in the MPA-pTHP-1 cells was increased, but its IL-10 production was suppressed by either the progesterone/glucocorticoid antagonist (Mifepristone) or MEK inhibitor (U0126). Taken together, MPA could drive monocyte differentiation toward an M2 phenotype that mimics decidual macrophages. This finding holds great potential to combat chronic endometrial inflammation.

Background Obesity is caused by excessive accumulation of body fat and is closely related to complex metabolic diseases. Raspberry ketone (RK), a major aromatic compound in red raspberry, was recently reported to possess anti-obesity effects. However, its mechanisms are unclear. Aim Adipogenesis plays a critical role in obesity and, therefore, this study aimed to investigate the effect and mechanisms of action of RK on adipogenesis in 3T3-L1 preadipocytes. Materials and methods 3T3-L1 preadipocytes were differentiated in medium containing insulin, dexamethasone, and 1-methyl-3-isobutylxanthine. Adipocyte lipid contents were determined using oil-red O staining while adipogenic transcription factor and lipogenic protein expressions were determined using western blotting. Results RK (300–400 µM) strongly inhibited lipid accumulation during 3T3-L1 preadipocyte differentiation into adipocytes. RK reduced the CCAAT/enhancer-binding protein-α (C/EBP-α), peroxisome proliferation-activated receptor-γ (PPAR-γ), fatty acid synthase (FAS), and fatty acid-binding protein 4 (FABP4) expressions and increased heme oxygenase-1 (HO-1), Wnt10b, and β-catenin expressions in 3T3-L1 adipocytes. Additionally, RK inhibited lipid accumulation, and adipogenic transcription factor and lipogenic protein expressions were all decreased by inhibiting HO-1 or β-catenin using tin protoporphyrin (SnPP) or β-catenin short-interfering RNA (siRNA), respectively. Furthermore, Wnt10b and β-catenin expressions were negatively regulation by SnPP. Conclusion RK may exert anti-adipogenic effects through modulation of the HO-1/Wnt/beta-catenin signaling pathway.

Aims Obesity is not only associated with metabolic diseases but is also a symptom of menopause in women. To date, there are no effective drugs for the management of obesity, and it is important to find new agents with fewer side effects, for the treatment of obesity. This study aimed to determine the anti-obesity effect of 17-dimethylaminoethylamino-17-demethoxygeldanamycin (17-DMAG), a heat shock protein 90 (Hsp90) inhibitor, and its underlying mechanism in rats with ovariectomy-induced obesity. Main methods Ovariectomy (Ovx) rats were treated with 17-DMAG (1 mg kg−1, intraperitoneally) for eight weeks from one week after surgery. The body weight, food intake, locomotor activity, adipogenic- and autophagy-related protein expression in white adipose tissue (WAT) and plasma triglyceride (TG) levels were measured in sham and Ovx rats. Key findings Compared with sham rats, Ovx rats showed increased weight gain, food intake, WAT mass, TG levels, adipogenic protein expression, and decreased locomotor activity. Furthermore, autophagy-related proteins and Foxo3a of WAT were significantly increased in Ovx rats. However, with the exclusion of increased food intake, the changes induced by Ovx were all reversed in 17-DMAG-treated Ovx rats. In addition, the expression of Hsp70 and phosphorylation of Akt increased in 17-DMAG-treated Ovx rats. Significance These results suggest that 17-DMAG significantly ameliorated obesity induced by Ovx, and this phenomenon is accompanied by the downregulation of adipogenic-related and autophagy-related proteins as well as the upregulation of Akt-phosphorylation and Hsp70 expression. Therefore, 17-DMAG may be a potential agent for preventing or treating obesity in postmenopausal women.

Obesity is a strong risk factor for the development of cardiovascular diseases and is associated with a marked increase in circulating leptin concentration. Leptin is a peptide hormone mainly produced by adipose tissue and is regulated by energy level, hormones and various inflammatory mediators. Genistein is an isoflavone that exhibits diverse health‐promoting effects. Here, we investigated whether genistein suppressed the atherogenic effect induced by leptin. The A10 cells were treated with leptin and/or genistein, and then the cell proliferation and migration were analysed. The reactive oxygen species (ROS) and proteins levels were also measured, such as p44/42MAPK, cell cycle‐related protein (cyclin D1 and p21) and matrix metalloproteinase‐2 (MMP‐2). Immunohistochemistry and morphometric analysis were used for the neointima formation in a rat carotid artery injury model. Genistein (5 μM) significantly inhibited both the proliferation and migration of leptin (10 ng/ml)‐stimulated A10 cells. In accordance with these finding, genistein decreased the leptin‐stimulated ROS production and phosphorylation of the p44/42MAPK signal transduction pathway. Meanwhile, genistein reversed the leptin‐induced expression of cyclin D1, and cyclin‐dependent kinase inhibitor, p21. Genistein attenuated leptin‐induced A10 cell migration by inhibiting MMP‐2 activity. Furthermore, the leptin (0.25 mg/kg)‐augmented neointima formation in a rat carotid artery injury model was attenuated in the genistein (5 mg/kg body weight)‐treated group when compared with the balloon injury plus leptin group. Genistein was capable of suppressing the atherogenic effects of leptin in vitro and in vivo, and may be a promising candidate drug in the clinical setting.

Heat shock protein 70 (Hsp70) preconditioning induces thermotolerance, and adenosine monophosphate (AMP)‐activated protein kinase (AMPK) plays a role in the process of autophagy. Here, we investigated whether 17‐dimethylaminoethylamino‐17‐demethoxy‐geldanamycin (17‐DMAG) protected against heat stroke (HS) in rats by up‐regulation of Hsp70 and phosphorylated AMPK (pAMPK). To produce HS, male Sprague–Dawley rats were placed in a chamber with an ambient temperature of 42°C. Physiological function (mean arterial pressure, heart rate and core temperature), hepatic and intestinal injury, inflammatory mediators and levels of Hsp70, pAMPK and light chain 3 (LC3B) in hepatic tissue were measured in HS rats or/and rats pre‐treated with 17‐DMAG. 17‐DMAG pre‐treatment significantly attenuated hypotension and organ dysfunction induced by HS in rats. The survival time during HS was also prolonged by 17‐DMAG treatment. Hsp70 expression was increased, whereas pAMPK levels in the liver were significantly decreased in HS rats. Following pre‐treatment with 17‐DMAG, Hsp70 protein levels increased further, and pAMPK levels were enhanced. Treatment with an AMPK activator significantly increased the LC3BII/LC3BI ratio as a marker of autophagy in HS rats. Treatment with quercetin significantly suppressed Hsp70 and pAMPK levels and reduced the protective effects of 17‐DMAG in HS rats. Both of Hsp70 and AMPK are involved in the 17‐DMAG‐mediated protection against HS. 17‐DMAG may be a promising candidate drug in the clinical setting.

Promoting white adipose tissue (WAT) to acquire brown-like characteristics is a promising approach for obesity treatment. Although raspberry ketone (RK) has been reported to possess antiobesity activity, its effects on the formation of brown-like adipocytes remain unclear. Therefore, we investigated the effects and underlying mechanism of RK on WAT browning in 3T3-L1 adipocytes and rats with ovariectomy (Ovx)-induced obesity. RK (100 μM) significantly induced browning of 3T3-L1 cells by increasing mitochondrial biogenesis and the expression of browning-specific proteins (PR domain containing 16, PRDM16; peroxisome proliferator-activated receptor gamma coactivator 1-alpha, PGC-1α; uncoupling protein-1, UCP-1) and lipolytic enzymes (hormone-sensitive lipase and adipose triglyceride lipase). RK significantly reduced the expression of the autophagy-related protein Atg12 and increased the expression of p62 and heme oxygenase 1 (HO-1). Additionally, these effects of RK were reversed by the HO-1 inhibitor SnPP (20 μM). In addition, RK (160 mg/kg, gavage, for 8 weeks) significantly reduced body weight gain (Ovx+RK, 191.8 ± 4.6 g vs. Ovx, 223.6 ± 5.9; P < .05), food intake, the amount of inguinal adipose tissue (Ovx+RK, 9.05 ± 1.1 g vs Ovx, 12.9 ± 0.92 g; P < .05) and the size of white adipocytes in Ovx rats. Moreover, compared to expression in the Ovx group, the levels of browning-specific proteins were significantly higher and the levels of autophagy-related proteins were significantly lower in the Ovx+RK group. Therefore, this study elucidated the mechanism associated with RK-induced WAT browning and thus provides evidence to support the clinical use of RK for obesity treatment.

Leucine-rich repeats and WD repeat domain containing protein 1 (LRWD1) is a testis-specific protein that mainly expressed in the sperm neck where centrosome is located. By using microarray analysis, LRWD1 is identified as a putative gene that involved in spermatogenesis. However, its role in human male germ cell development has not been extensively studied. When checking in the semen of patients with asthenozoospermia, teratozoospermia, and asthenoteratozoospermia, the level of LRWD1 in the sperm neck was significantly reduced with a defective neck or tail. When checking the sub-cellular localization of LRWD1 in the cells, we found that LRWD1 resided in the centrosome and its centrosomal residency was independent of microtubule transportation in NT2/D1, the human testicular embryonic carcinoma, cell line. Depletion of LRWD1 did not induce centrosome re-duplication but inhibited microtubule nucleation. In addition, the G1 arrest were observed in LRWD1 deficient NT2/D1 cells. Upon LRWD1 depletion, the levels of cyclin E, A, and phosphorylated CDK2, were reduced. Overexpression of LRWD1 promoted cell proliferation in NT2/D1, HeLa, and 239T cell lines. In addition, we also observed that autophagy was activated in LRWD1 deficient cells and inhibition of autophagy by chloroquine or bafilomycin A1 promoted cell death when LRWD1 was depleted. Thus, we found a novel function of LRWD1 in controlling microtubule nucleation and cell cycle progression in the human testicular embryonic carcinoma cells. J. Cell. Biochem. 119: 314-326, 2018. © 2017 Wiley Periodicals, Inc.

Purpose LRWD1 is a protein that contains LRR and WDs domains and is important in regulating spermatogenesis. However, the roles of LRR or WDs domains in the expression of LRWD1 remain unclear. Materials and methods The NT2/D1 cells separately transfected with full length of LRWD1 gene ( LRWD WT ) or genes with deleted sequences in the LRR domain ( LRWD1 ΔLRR ), WD1 domain ( LRWD1 ΔWD1 ), WD2 domain ( LRWD1 ΔWD2 ), WD3 domain ( LRWD1 ΔWD3 ) and entire three WD domains ( LRWD1 Δ3×WD ) were applied to investigate the expression levels of LRWD1 protein by either Western blot or flow cytometry. The associated proteins in these mutated LRWD1 proteins were identified by mass spectrometry. Results Deletion of the LRR domain significantly decreased the expression of LRWD1 protein. With the treatment of MG132, the LRR domain may functions in preventing LRWD1 protein from proteasome-mediated degradation. In the co-immunoprecipitation analysis, protein receptor of tumor necrosis factor 2 (TNFR2) was specifically observed to be associated with LRR-deficient LRWD1 protein. Conclusions The LRR domain is significantly correlated to the stability of LRWD1 protein. Determining if the stability is modulated by TNFR2 is worthy of further study.

The health care sector has become increasingly interested in developing personal health record (PHR) systems as an Internet-based telehealthcare implementation to improve the quality and decrease the cost of care. However, the factors that influence patients' intention to use PHR systems remain unclear. Based on physicians' therapeutic expertise, we implemented a web-based infertile PHR system and proposed an extended Technology Acceptance Model (TAM) that integrates the physician-patient relationship (PPR) construct into TAM's original perceived ease of use (PEOU) and perceived usefulness (PU) constructs to explore which factors will influence the behavioral intentions (BI) of infertile patients to use the PHR. From ninety participants from a medical center, 50 valid responses to a self-rating questionnaire were collected, yielding a response rate of 55.56%. The partial least squares (PLS) technique was used to assess the causal relationships that were hypothesized in the extended model. The results indicate that infertile patients expressed a moderately high intention to use the PHR system. The PPR and PU of patients had significant effects on their BI to use PHR, whereas the PEOU indirectly affected the patients' BI through the PU. This investigation confirms that PPR can have a critical role in shaping patients' perceptions of the use of healthcare information technologies. Hence, we suggest that hospitals should promote the potential usefulness of PHR and improve the quality of the physician-patient relationship to increase patients' intention of using PHR.

The purpose of this study was to describe the experiences of pregnancy of women in Taiwan after undergoing at least three cycles of assisted reproductive technology (ART) over a period exceeding 3 years. Fifteen previously infertile female patients diagnosed with primary infertility participated in this study within 1 year of delivering a baby. Participants were between the ages of 31 and 44 with an average age of 39.41. They had been married an average of 11.33 years and had undergone an average of 5.25 years of infertility treatment. A phenomenological qualitative method with in-depth interviews was employed for the collection of data. Our findings reveal that the safety and health of the fetus is the primary concern of previously infertile pregnant mothers. Other concerns include physical/physiological changes, psychosocial reactions, the transition of identity during pregnancy, insights gained through pregnancy and labor, and the impact of Taiwanese society on the pregnancy. The post-infertility pregnant women in this study endured a great deal to conceive through ART, safeguarded the health of their fetus, and managed their reactions. Practitioners of couple and family counseling should offer assistance to post-infertility pregnant women through psychological counseling and consultation to help them deal with their biopsychosocial reactions and identity transition.

Magnolol is a compound extracted from the Chinese medicinal herb Magnolia officinalis. It has multiple pharmacological effects, notably as an anti-oxidant. The aim of this study was to evaluate the effects of magnolol on sepsis induced by intravenous (i.v.) administration of lipopolysaccharide (LPS; 10 mg/kg) in anaesthetized Wistar rats. Magnolol (4 microg/kg, i.v.) was administered at 30 min after LPS injection. Post-treatment with magnolol significantly attenuated the deleterious haemodynamic changes (e.g., hypotension and bradycardia) caused by LPS. Meanwhile, magnolol significantly inhibited the elevation of plasma levels of tumor necrosis factor alpha, glutamate-oxaloacetate transaminase, glutamate-pyruvate transaminase and blood urine nitrogen caused by LPS. The induction of inducible nitrous oxide (NO) synthase and the overproduction of NO and superoxide anions by LPS were also significantly reduced by post-treatment with magnolol. Moreover, the plasma level of the thrombin-antithrombin complex following administration of LPS was also reduced by post-treatment with magnolol. Thus, the beneficial effects of magnolol on LPS-induced sepsis result from its anti-inflammatory, anti-coagulatory, and anti-oxidant effects.

Objective: To assess the effects of 2 months of treatment with Femarelle for climacteric syndrome in Taiwanese postmenopausal women. Materials and methods: A multi-center, open-label trial of 260 postmenopausal women, age ≥ 45 years with vasomotor symptoms. Women were enrolled after obtaining a detailed medical history and a thorough physical examination. They then received Femarelle (640 mg/d) twice daily for 8 weeks. The primary outcome was the changes in the frequency and severity of hot flushes from baseline to 4 weeks (1 month) and 8 weeks (2 months). Changes of general climacteric syndrome were assessed using a modified climacteric scale designed by Greene. Results: The frequency and severity of hot flushes were significantly improved with Femarelle use (p

To compare the clinical outcome of IVF treatment after pituitary suppression with two different oral contraceptives (OCs).65 patients who received IVF treatment was classified into 2 groups based on the difference of OCs they used for pituitary suppression before ovarian hyperstimulation. Group 1 included 36 patients who received monophasic OCs. Group 2 included 29 patients who received triphastic OCs. Both groups received the OCs from the 5th day of the cycle for consecutive 21 days. The hormone profiles after OCs and clinical outcome of IVF treatment were compared between two groups. Two-sample t-tests and X2 tests were used for statistical analyses. P0.05 was considered statistically significant.The mean age and basal hormone profiles were comparable between two groups. After ovulation suppression with different OCs, the day 2 FSH and LH value revealed statistically significant difference between two groups(4.2+/-1.8 vs 6.0+/-2.6; 2.7+/-2.0 vs 4.2+/-3.3 respectively). The numbers of oocyte per retrieval and fertilization rate were comparable between two groups, but higher quality embryos as revealed by the cleavage speed were noted in the triphastic OCs group. Although statistically not significant, higher implantation rate and pregnancy rate were also noted in the triphastic OCs group.Different OCs for pituitary suppression can result in different hormone profiles. Ovulation induction in IVF treatment should be individualized according to these hormone changes to achieve the optimal clinical outcome. Triphastic OCs exceeds monophastic OCs in producing good quality embryo in IVF-ET treatment.

Objective: Human umbilical cord blood cells (HUCBCs) are effective in the treatment of conventional stroke in experimental models. In the study described herein, we administered HUCBCs into the femoral vein or directly into the cerebral ventricular system and assessed their effects on circulatory shock, cerebral ischemia, and damage during heatstroke. Design: Controlled, prospective study. Setting: Hospital medical research laboratory. Subjects: Sprague-Dawley rats (287 16 g body weight, males). Interventions: Anesthetized rats, immediately after the onset of heatstroke, were divided into four major groups and given the following: a) normal saline or AIM-V medium intravenously (0.3 mL) or intracerebroventricularly (10 L); b) peripheral blood mononuclear cells (5 10 6 in 0.3 mL AIM-V medium, intravenously, or 5 10 5 in 10 L AIM-V medium, intracerebroventricularly); or c) HUCBCs (5 10 6 in 0.3 mL AIM-V medium, intravenously, or 5 10 5 in 10 L AIM-V medium, intracerebroventricularly). Another group of rats, under urethane anesthesia, were exposed to room temperature (26°C) and used as normothermic controls. Urethane-anesthetized animals were exposed to an ambient temperature of 43°C to induce heatstroke. Their physiologic and biochemical parameters were continuously monitored. Measurements and Main Results: When the vehicle-treated rats underwent heat exposure, their survival time values were found to be 21–23 mins. Resuscitation with intravenous or intracerebroventricular doses of HUCBCs, but not peripheral blood mononuclear cells, immediately at the onset of heatstroke significantly improved survival during heatstroke (61–148 mins). As compared with values for normothermic controls, the vehicletreated heatstroke rats had lower mean arterial pressure, cerebral blood flow, and brain PO2 values but higher intracranial pressure and cerebral ischemia values and more injury markers. The circulatory shock, intracranial hypertension, cerebral hypoperfusion and hypoxia, increment of cerebral ischemia, and damage markers during heatstroke were all significantly attenuated by intravenous or intracerebroventricular delivery of HUCBCs but not peripheral blood mononuclear cells. Conclusions: We successfully demonstrate that HUCBC therapy may resuscitate heatstroke victims by reducing circulatory shock and cerebral ischemic injury; central delivery of HUCBCs seems superior to systemic delivery of HUCBCs in resuscitating patients with heatstroke. (Crit Care Med 2005; 33:1377–1383)

Significant follicle loss from frozen ovarian grafts is unavoidable. The authors evaluated the protective effects of the antiapoptotic agent sphingosine-1-phosphate (S1P) on vitrified ovarian grafts. Three-week-old sexually immature female FVB mice were divided into 4 groups, fresh, control without S1P, 0.5 mmol/L S1P, and 2 mmol/L S1P. The ovaries were pretreated with S1P for 1 hour and then cryopreserved by modified vitrification. The frozen–thawed ovaries were autotransplanted under the back muscles of mice for 10 days. Expression of apoptosis-related genes encoding caspase 3 and c-Myc was analyzed in the vitrified ovaries and 10 days after transplantation using real-time quantitative polymerase chain reaction. To quantify the ovarian reserve, anti-Mullerian hormone (AMH) levels and follicles were measured in the 10-day vitrified ovarian grafts. Caspase 3 and c-Myc messenger RNA did not differ significantly in the 4 groups after vitrification but was significantly upregulated in the control group after transplantation. The AMH levels and primordial follicle pool were significantly higher in the S1P-treated groups than in the control group but lower than that in the fresh group. The S1P protects vitrified ovarian grafts from ischemic reperfusion injury rather than from vitrification-associated process.

Neurodegenerative brain disorders such as Alzheimer's disease (AD) have been well investigated. However, significant methods for the treatment of the progression of AD are unavailable currently. Heat shock protein 70 (Hsp70) plays important roles in neural protection from stress by assisting cellular protein folding. In this study, we investigated the effect and the molecular mechanism of YC-1, an activator of guanylyl cyclase (GC), on Aβ25-35-induced cytotoxicity in differentiated PC12 cells. The results of this study showed that Aβ25-35 (10 µM) significantly increased p25 protein production in a pattern that was consistent with the increase in μ-calpain expression. Moreover, Aβ25-35 significantly increased tau hyperphosphorylation and induced differentiated PC12 cell death. YC-1 (0.5-10 µM) prevented the cell death induced by Aβ25-35. In addition, YC-1 (1, 10 µM) significantly blocked Aβ25-35-induced μ-calpain expression and decreased the formation of p25 and tau hyperphosphorylation. Moreover, YC-1 (5-20 µM) alone or combined with Aβ25-35 (10 µM) significantly increased the expression of Hsp70 in differentiated PC12 cells. The neuroprotective effect of YC-1 was significantly attenuated by an Hsp70 inhibitor (quercetin, 50 µM) or in PC12 cells transfected with an Hsp70 small interfering RNA. However, pretreatment of cells with the GC inhibitor ODQ (10 µM) did not affect the neuroprotective effect of YC-1 against Aβ25-35 in differentiated PC12 cells. These results suggest that the neuroprotective effect of YC-1 against Aβ25-35-induced toxicity is mainly mediated by the induction of Hsp70. Thus, YC-1 is a potential agent against AD.

We tested differences in serum apelin levels between women with polycystic ovary syndrome (PCOS) and those with a healthy regular menstrual cycle, finding that apelin levels were higher in normal women and that apelin was positively correlated with apolipoprotein A levels.

Objective: Adenosine monophosphate-activated protein kinase (AMPK) acts as a cellular energy sensor, being activated during states of low energy charge. Hypothalamic AMPK is altered by hormonal and metabolic signals and mediates the feeding response. The aims of this study were to examine whether the phosphorylation of AMPKα in the hypothalamus is affected by ovariectomy (Ovx) and thus would be involved in the development of obesity in rats. Methods: Body weight, food intake, hypothalamic phosphorylated AMPKα (pAMPKα) protein expression, and plasma leptin and adiponectin levels were measured in female rats after either Ovx or sham operations. These patterns were also observed after treatment with 17β-estradiol, compound C, and leptin in Ovx rats. Results: Compared with control rats, Ovx led to increased body weight and food intake at 2 to 8 weeks after operation. Meanwhile, plasma leptin and adiponectin levels and hypothalamic pAMPKα expression were significantly increased after Ovx. Replacement of estradiol significantly reversed these effects. Treatment with compound C, an AMPKα inhibitor, for 1 week produced a reduction in food intake, body weight, and plasma leptin and adiponectin levels. Meanwhile, these effects were reversed upon withdrawal of compound C. In addition, central injection of leptin also significantly reduced body weight, food intake, plasma leptin and adiponectin levels, and hypothalamic pAMPKα expression relative to those of the Ovx group. Conclusions: Increased hypothalamic pAMPKα expression may contribute to hyperphagia during the development of Ovx-induced obesity in rats.

Background/Aims: Retinol-binding protein 4 (RBP4) has been shown to be associated with insulin resistance and fatty acid metabolism. We hypothesize that RBP4 might play a role in fetal growth and that cord serum RBP4 may act as a marker of fetal growth, independent of fetal insulin levels. Methods: Twenty-one women having fetuses in the top quartile (>75th percentile) of birth weights for gestational age were enrolled into the trial, along with 21 women having fetuses in the bottom quartile (Results: Cord serum RBP4 and insulin concentrations were significantly higher in the top quartile group (14.3 ± 3.7 ng/ml, 3.8 ± 5.2 µIU/ml) than in the bottom quartile group (11.3 ± 2.6 ng/ml, 0.9 ± 1.4 µIU/ml; p = 0.004, p = 0.017). Cord serum RBP4 and insulin as well as gestational age (r = 0.744, r2 = 0.553, p < 0.001) were significantly correlated with fetal birth weights. Conclusion: Cord serum RBP4 concentrations were higher in subjects with top quartile fetuses than in those with bottom quartile fetuses. Cord serum RBP4 concentrations were significantly correlated with fetal birth weight. These findings may indicate that cord serum RBP4 plays a regulatory role in fetal growth.

Objective The aim of this study was to identify an optimal stimulation protocol for intrauterine insemination (IUI) to obtain an acceptable pregnancy rate and low frequency of multiple pregnancies. Materials and Methods In total, 340 patients, who received intrauterine insemination because of ovulation dysfunction, were enrolled in this study. Group I consisted of 203 patients who received recombinant FSH (r-FSH) 150U every other day as an ovulation induction agent. Group II consisted of 137 patients who received r-FSH 100U every other day as an ovulation induction agent. All patients in both groups also received clomiphene citrate 100 mg/day for consecutive five days from the fifth day of the cycle. Only patients with at least two follicles >18 mm on the human chorionic gonadotropin injection days were included in this study. Results The clinical pregnancy rate was 14.8% (30/203) in Group I compared with 20.4% (28/137) in Group II, p > 0.05. The incidence of multiple pregnancy was 41.7% (10/24) in Group I compared with 12.5% (3/24) in Group II, p Conclusions The concurrent use of low-dose r-FSH and clomiphene citrate would seem not only to be cost-effective but also highly satisfactory in that it prevents high-order multiple pregnancies.

Aim: To complete comprehensive haplotype analysis of USP26 for both fertile and infertile men. Methods: Two hundred infertile men with severe oligospermia or non-obstructive azoospermia were subjected to sequence analysis for the entire coding sequences of the USP26 gene. Two hundred men with proven fertility were genotyped by primer extension methods. Allele/genotype frequencies, linkage disequilibrium (LD) characteristics and haplotypes of fertile men were compared with infertile men. Results: The allele frequencies of five single nucleotide polymorphisms (370-371insACA, 494T>C, 576G>A, ss6202791C>T, 1737G>A) were significantly higher in infertile patients than control subjects. The major haplotypes in infertile men were TACCGA (28% of the population), TGCCGA (15%), TACCAA (8%), TGCCAA (6%), TATCAA (5%) and CATCAA (5%). The major haplotypes for the control subjects were TACCGA (58% of the population), CACCGA (7%), CATCGA (6%) and TGCCGA (5%). Haplotypes TGCCGA, TATCAA, CATCAA, CATCGC, TACCAA and TGCCAA were over-transmitted in patients with spermatogenic defect, whereas haplotypes TACCGA, CACCGA, and CATCGA were under-transmitted in these patients. Conclusion: Some USP26 alleles and haplotypes are associated with spermatogenic defect in the Han nationality in Taiwan, China. (Asian J Androl 2008 Nov; 10: 896–904)

Objective To test whether there was a difference in serum retinol-binding protein 4 (RBP4) levels between subjects with polycystic ovary syndrome (PCOS) and those with a healthy regular menstrual cycle and, in addition, to correlate serum RBP4 levels with a variety of parameters. Design Clinical study. Setting University hospital. Patient(s) A total of 74 nonobese women were evaluated. Thirty-seven had PCOS, whereas the remaining 37 served as control subjects. Intervention(s) Serum RBP4 levels were analyzed using ELISA. Main Outcome Measure(s) Serum levels of FSH, LH, TSH, E2, T, insulin, glucose, cholesterol, triglycerides, and RBP4. Result(s) The women with PCOS had higher levels of serum RBP4, waist-to-hip ratio, LH, T, insulin, homeostatic model assessment of insulin resistance, cholesterol, and triglycerides. Logistic regression analyses revealed a significant association between odds ratio (OR) values of PCOS and both T (OR = 1.125; 95% confidence interval [CI] 1.050–1.205), and cholesterol levels (OR = 1.029; 95% CI 1.004–1.056). Age and triglycerides were significantly correlated to serum RBP4 levels by multiple linear regression analysis. Conclusion(s) Our study has shown that [1] elevated RBP4 levels might arise from triglyceride metabolism, and that RBP4 levels might not be influenced by PCOS itself. [2] RBP4 might not be a useful marker of insulin resistance in subjects with PCOS.

Leptin is a peptide hormone, which has a central role in the regulation of body weight; it also exerts many potentially atherogenic effects. Ferulic acid ethyl ester (FAEE) has been approved for antioxidant properties. The aim of this study was to investigate whether FAEE can inhibit the atherogenic effects of leptin and the possible molecular mechanism of its action. Both of cell proliferation and migration were measured when the aortic smooth muscle cell (A10 cell) treated with leptin and/or FAEE. Phosphorylated p44/42MAPK, cell cycle-regulatory protein (for example, cyclin D1, p21, p27), β-catenin and matrix metalloproteinase-9 (MMP-9) proteins levels were also measured. Results demonstrated that leptin (10, 100 ng ml(-1)) significantly increased the proliferation of cells and the phosphorylation of p44/42MAPK in A10 cells. The proliferative effect of leptin was significantly reduced by the pretreatment of U0126 (0.5 μM), a MEK inhibitor, in A10 cells. Meanwhile, leptin significantly increased the protein expression of cyclin D1, p21, β-catenin and decreased the expression of p27 in A10 cells. In addition, leptin (10 ng ml(-1)) significantly increased the migration of A10 cells and the expression of MMP-9 protein. Above effects of leptin were significantly reduced by the pretreatment of FAEE (1 and 10 μM) in A10 cells. In conclusion, FAEE exerts multiple effects on leptin-induced cell proliferation and migration, including the inhibition of p44/42MAPK phosphorylation, cell cycle-regulatory proteins and MMP-9, thereby suggesting that FAEE may be a possible therapeutic approach to the inhibition of obese vascular disease.

This follow-up study examined the major complications among 4307 operative gynecologic laparoscopies. The overall complication rate and each individual category were compared with those of our previous study period. The clinical outcome and salvage procedures were correlated with the time of recognition and the severity of initial procedures in the individual injury type.Retrospective, comparative study based on medical record reviewing (Canadian Task Force classification II-3).Tertiary teaching hospital, Chi Mei Foundation Hospital in southern Taiwan.Records of women (n = 4307) aged 40.5 +/- 11.7 years (mean +/- SD [95% CI 40.1-40.5]) who underwent operative gynecologic laparoscopies from January 2000 through February 2006 were reviewed in this study. The complications were compared with those of our previous study based on 1507 laparoscopies performed between December 1992 and November 1999 for follow-up comparison.Gynecologic laparoscopic surgeries.Thirty-four complications occurred in 31 patients requiring repair procedures, 3 of whom had multiple complications, with an overall complication rate of 0.72% (31/4307). There were 13 bladder injuries (0.30%), 7 bowel injuries (0.16%), 3 cases of internal bleeding (0.07%), 4 vaginal stump hematomas or abscesses (0.09%), 3 ureteral injuries (0.07%), 3 major vessel injuries (0.07%), and 1 trocar site hematoma (0.02%). In addition, there were 125 (2.88%) postoperative blood transfusions without additional operative intervention. The major complication rate decreased compared with that of the previous study (0.72% [95% CI 0.51%-1.02%] vs 1.59% [95% CI 1.07%-2.36%]; p = .005). The overall complication rates were not significantly different between laparoscopic hysterectomy (LH) group and non-LH group. However, bladder injury happened more frequently in the LH group, whereas bowel injury was more common in the non-LH group. In addition, the severity of the original injury, timing of recognition, and accompanying salvage procedures correlated with the clinical outcomes.The significantly decreased major complication rate, as compared with that of our previous study period, confirms the importance of experience accumulation and use of preventive maneuvers in reducing the complication rate. There were no significant differences among the individual injury category during these 2 study periods. The manifestations of bowel injury were highly variable and individualized. The accumulation of surgical experience with the aid of preventive maneuvers is helpful to reduce the complication rate significantly.

Objective To test the diagnostic efficiency of a gene-specific, five-marker screening strategy for the detection of Y chromosome deletions. Design Prospective case study. Setting University genetics laboratory and reproductive clinics. Patient(s) Six hundred twenty-seven infertile men and 212 fertile men. Intervention(s) Peripheral blood samples were screened for Y chromosome deletions in a triple-blind fashion using three protocols: protocol I consisted of five gene-specific markers, including USP9Y, DBY, SMCY, RBM1 , and DAZ ; protocol II included 14 gene-specific markers; and protocol III consisted of six sequence-tagged sites (STSs) markers recommended by EAA/EMQN. Main Outcome Measure(s) Deletion status of Y chromosome genes or sequence-tagged sites. Result(s) Protocols I and II identified the same 41 infertile patients with Y deletions. Protocol III identified 38 infertile patients with Y deletions, and all 38 patients were also identified by protocols I and II. One patient with isolated USP9Y deletion and two patients with isolated DBY deletions, as detected by protocols I and II, could not be identified by protocol III. Conclusion(s) We observed mostly consistent results between our protocols and the EAA/EMQN protocol. This gene-specific, five-marker screening panel provides the same diagnostic efficiency as the EAA/EMQN protocol and may be considered an alternative to the EAA/EMQN protocol.

Objective. To elucidate whether there is a distention of the symphysis pubis intra partum and whether a history of pelvic pain during pregnancy is associated with increased distention during delivery.Methods. Using ultrasonography, the width of the symphysis pubis was measured on two occasions during delivery; firstly, upon engagement of the fetal head in the pelvic inlet, secondly, when the largest diameter of the fetal head was at the level of the ischiadic spines. The first measurement was conducted in 24 patients, the second measurement was obtained in 16 of these patients.Results. The average width of the symphysis pubis at onset of labor was 5.8 mm. The average distention of the symphysis during labor was 1.1 mm for those without pelvic pain and 0.2 mm for those with a history of pelvic pain during pregnancy (p=0.02). Parity and ensuing birth weight had no statistically significant influence on degree of distention.Conclusion. The symphyseal distention during labor is minimal regardless of parity an...