Novel variations in TENT5D lead to teratozoospermia in infertile patients.

PURPOSE: Teratozoospermia is the main pathogenic factor of male infertility. However, the genetic etiology of teratozoospermia is largely unknown. This study aims to clarify the relationship between novel variations in TENT5D and teratozoospermia in infertile patients. MATERIALS AND METHODS: Two infertile patients were enrolled. Routine semen analysis of patients and normal controls was conducted with the WHO guidelines. Whole-exome sequencing (WES) was conducted to identify pathogenic variants in the two patients. Morphology and ultrastructure analysis of spermatozoa in the two patients was determined by Papanicolaou staining, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The functional effect of the identified variants was analyzed by immunofluorescence staining and western blotting. The expression of TENT5D in different germ cells was detected by immunofluorescence staining. RESULTS: Two new hemizygous variations, c.101C > T (p.P34L) and c.125A > T (p.D42V), in TENT5D were detected in two patients with male infertility. Morphology analysis showed abnormalities in spermatozoa morphology in the two patients, including multiple heads, headless, multiple tails, coiled, and/or bent flagella. Ultrastructure analysis showed that most of the spermatozoa exhibited missing or irregularly arranged '9+2' structures. Further functional experiments confirmed the abrogated TENT5D protein expression in patients. In addition, both p.P34L and p.D42V substitutions resulted in a conformational change of the TENT5D protein. We precisely analyzed the subcellular localization of TENT5D in germ cells in humans and mice. And we found that TENT5D was predominantly detected in the head and flagellum of elongating spermatids and epididymal spermatozoa. CONCLUSIONS: Our results showed further evidence of a relationship between TENT5D mutation and human male infertility, providing new genetic insight for use in the diagnosis and treatment of male infertility.

Evaluation of the respective contribution of anode and cathode for triclosan degradation in a bioelectrochemical system.

In this work, the bioelectrochemical system (BES) is a feasible alternative for successfully degrading typical refractory emerging contaminant triclosan (TCS). A single-chamber BES reactor with an initial TCS concentration of 1 mg/L, an applied voltage of 0.8 V, and a solution buffered with 50 mM PBS degraded 81.4 ± 0.2% of TCS, exhibiting TCS degradation efficiency improvement to 90.6 ± 0.2% with a biocathode formed from a reversed bioanode. Both bioanode and biocathode were able to degrade TCS with comparable efficiencies of 80.8 ± 4.9% and 87.3 ± 0.4%, respectively. Dechlorination and hydrolysis were proposed as the TCS degradation pathway in the cathode chamber, and another hydroxylation pathway was exclusive in the anode chamber. Microbial community structure analysis indicated Propionibacteriaceae was the predominant member in all electrode biofilms, and the exoelectrogen Geobacter was enriched in anode biofilms. This study comprehensively revealed the feasibility of operating BES technology for TCS degradation.

Mathematical processing of trading strategy based on long short-term memory neural network model.

At present, gold and bitcoin have become mainstream assets in market transactions. Due to the volatility of gold and bitcoin prices, we can buy and sell assets like gold and bitcoin the same way we buy and sell stocks. The research goal of this article is to develop an optimal trading strategy that maximizes our post-trade returns. By studying the relationship between the two, on the one hand, it supplements and enriches the theoretical research on the rate of return of gold and Bitcoin, on the other hand, it provides a certain reference for investors to construct investment strategies. The research on the cointegration relationship between them has important practical significance. At the same time, it has important practical significance for the research on the cointegration relationship between bitcoin and gold.

MicroRNA­187 is an independent prognostic factor in lung cancer and promotes lung cancer cell invasion via targeting of PTRF.

MicroRNAs (miRNAs) are involved in the progression of different types of cancers giving new hope for cancer treatment. The role and regulatory mechanism of microRNA­187 (miR­187) are largely unknown. In the present study, 74 patients with non­small cell lung cancer (NSCLC) were selected. Tumor tissues and matched normal tissues were collected for determining the expression level of miR­187. Cell research was performed to detect the function of miR­187. The expression level was measured and miR­187 was found to be overexpressed in the NSCLC cell lines and tissues. Overexpression of miR­187 promoted cell proliferation in the A549 and H1650 cell lines. Moreover, overexpression of miR­187 also promoted cell migration and invasion. Polymerase I and transcript release factor (PTRF) was identified as a target of miR­187. Overexpression of miR­187 suppressed the expression of PTRF. Knockdown of PTRF promoted lung cancer cell invasion, and overexpression of PTRF had a negative effect on lung cancer cell invasion. The PTRF messenger RNA (mRNA) levels in cancer tissues were significantly lower than those in their adjacent normal lung tissues as determined by real­time PCR (RT­PCR). The expression of the PTRF protein was significantly weaker than that in the adjacent normal lung tissues using immunohistochemical staining. The findings revealed that miR­187 promotes cell growth and invasion by targeting PTRF and miR­187 may be a new prognostic factor for NSCLC.

Amlodipine prevents adriamycin-induced toxicity in cultured rat mesangial cells by up-regulation of Smad6, Smad7 expression.

Extensive studies have demonstrated that transforming growth factor-beta (TGF-ß) plays an important role in the progression of renal diseases. A central component of TGF-ß is the TGF-ß family-specific Smad signal transduction pathway. TGF-ß signals through Smad2, 4 to mediate renal fibrosis, whereas induction of Smad6, 7 inhibits renal fibrosis and inflammation. Amlodipine is the most frequently used antihypertensive drug among dihydropyridines. It is beneficial to the kidney and is widely used in treating kidney diseases. The aim of this study was to investigate effects of amlodipine on adriamycin-induced changes of lactate dehydrogenase (LDH) and expression of Smad6, 7 in rat mesangial cells. Results showed that amlodipine (10(-8) to 10(-5)mol/l) significantly decreased LDH activity in rat mesangial cells when given in combination with TGF-ß1 (P<0.01); amlodipine (10(-7), 10(-6)mol/l) significantly increased Smad6, 7 mRNA and protein expression in cells treated with adriamycin and TGF-ß1 (P<0.01). In conclusion, amlodipine protects against adriamycin-induced toxicity in rat mesangial cells by up-regulation of Smad6, 7 expressions.

Ligustrazine inhibits lipopolysaccharide-induced proliferation by affecting P27, Bcl-2 expression in rat mesangial cells.

Ligustrazine has a renoprotective effect against nephritis. In the present study, we investigated the roles of ligustrazine on lipopolysaccharide-induced changes of proliferation, cell cycle in cultured rat mesangial cells. 3-(4,5-dimethyltiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay revealed that rat mesangial cells treated with lipopolysaccharide (10mg/l) underwent significant proliferation compared with control group. This effect was significantly inhibited by ligustrazine (400 to 2500 mg/l). Flow cytometric analysis revealed that cells treated with lipopolysaccharide showed significant reduction in the ratio of G0/G1 phase and significant elevation in the ratio of S+G2/M phase. The changes of cell cycle induced by lipopolysaccharide were reversed by ligustrazine. In addition, lipopolysaccharide suppressed P27 protein expression was significantly increased by ligustrazine (100, 500, 2500 mg/l). Moreover, rat mesangial cells treated with lipopolysaccharide showed scanty apoptosis with up-regulation of Bcl-2expression, while Bax protein expression was not changed. Ligustrazine (100, 500, 2500 mg/l) significantly reversed lipopolysaccharide-induced up-regulation of Bcl-2 protein and increased apoptotic cell death. In summary, ligustrazine displayed a significant inhibiting effect on lipopolysaccharide-induced proliferation through increasing P27 and decreasing Bcl-2 protein expression in rat mesangial cells.

Differential roles of dihydropyridine calcium antagonist nifedipine, nitrendipine and amlodipine on gentamicin-induced renal tubular toxicity in rats.

In the present study, we investigated the antioxidative potencies of dihydropyridine calcium antagonists prototype nifedipine, the second generation drug nitrendipine, and the long acting, third generation drug amlodipine on gentamicin-induced renal tubular toxicity in Sprague-Dawley rats. In addition, we analyzed the relationship between renal tubular cell apoptosis and the antioxidative properties of these dihydropyridine calcium antagonists. Results showed that treatment with gentamicin alone caused significant changes in the levels of urinary protein, urinary N-acetyl-beta-d-glucosaminidase, serum creatinine, and blood urea nitrogen. Nifedipine and amlodipine effectively reversed the effect of gentamicin on these parameters. In contrast, nitrendipine either had no effect or worsened gentamicin-induced changes in the levels of urinary protein, urinary N-acetyl-beta-d-glucosaminidase, serum creatinine, and blood urea nitrogen. Furthermore, gentamicin treatment caused significant increases in the levels of malondialdehyde, nitric oxide, nitric oxide synthase and significant decreases in the levels of reduced glutathione, glutathione-S-transferase, and superoxide dismutase in kidney tissues. These effects were dramatically reduced by nifedipine and amlodipine but not affected by nitrendipine. In addition to the biochemical changes, histopathological studies showed that gentamicin caused structural damages in the kidneys; renal tubular cell apoptosis, a decrease in Bcl-2 expression and an increase in Bax expression were observed in all rats treated with gentamicin, nifedipine and amlodipine effectively reversed the effect of gentamicin while nitrendipine worsened them. In conclusion, this study clearly indicated that nifedipine and amlodipine protected against gentamicin-induced nephrotoxicity while nitrendipine had little effect, or even worsened.

[Construction of eukaryotic expression recombinant plasmid of trichomonas vaginalis ferredoxin gene].

Total DNA was extracted from T. vaginalis with Chelex-100 method and used as templates for PCR. The ferredoxin gene was directionally cloned into plasmid pMD-18T vector and subcloned into eukaryotic expression vector pcDNA3. 1 (+). The transformants were screened and identified by PCR and restriction analysis. The size of amplified ferredoxin gene was 306bp and the DNA sequence of cloned gene was same with that published.

[Molecular and chemical mechanism of Trichomonas vaginalis-induced contact-dependent cytotoxicity].

Trichomonas vaginalis parasitizes in human genitourinary tract. The protozoon adhering to target cell plays a critical role in its contact-dependent cytotoxicity. The enzymes synthesized by T.voginalis can hurt vaginalis epithelial cells (VECs) directly. The focal immune reaction in the location parasitized by the parasite may provide an immunologic protection. Meanwhile, inflammatory factors and immune cells may aggravate the situation. In general, the T. vaginalis-induced contact-dependent cytotoxicity is a result of the involvement of some molecular and chemical factors.

[Cloning and sequence analysis of Trichomonas vaginalis ferredoxin gene].

OBJECTIVE: To construct a recombinant plasmid containing ferredoxin gene of Trichomonas vaginalis. METHODS: Total DNA was extracted from Trichomonas vaginalis with Chelex-100 method and used as templates for PCR. Primers were designed based on the published sequence of the ferredoxin gene and used to amplify the Trichomonas vaginalis gene using PCR method. The ferredoxin gene obtained by PCR technique was directionally cloned into plasmid pMD-18T simple vector. The constructed recombinant plasmid was transferred into E. coli JM109. The transformants were screened and identified by PCR and restriction analysis. The DNA sequence of the gene was determined by Sanger's method. RESULTS: The size of amplified ferredoxin gene was 306bp. The correct recombinant plasmid was isolated and confirmed by PCR and restriction analysis. The DNA sequence of cloned gene was the same as the published sequence. CONCLUSION: The ferredoxin gene was successfully amplified and cloned into plasmid pMD-18T simple vector. The cloned ferredoxin gene could be used to produce recombinant protein and for study of its function.