The relationship between serum and seminal plasma vitamin D and male reproductive function / 医学研究生学报

Objective Male infertility accounts for 40 to 50% of the total number of infertility in the world. Among many factors that cause male infertility, vitamin D is considered to be directly related to male fertility. The purpose of this study was to explore the relationship between serum and seminal plasma vitamin D and male reproductive function, and provide a more comprehensive research direction for studying the specific mechanism of vitamin D on male reproduction. Methods A total of 198 infertile males, receiving andrological examination from June 2017 to January 2018 in the Center for Reproductive Medicine, Jinling Hospital (Nanjing, China) was included in our study. Serum and seminal plasma vitamin D levels were measured by electrochemiluminescence immunoassay (ECLIA) kits. The associations between vitamin D and biomarkers of male reproduction were analyzed. Results Serum 25(OH) vitamin D level [26.17(19.61-31.99)ng/mL] was in positive relation with semen volume[3.8(3.1-4.8)ng/mL] (r=0.229,P=0.003). Seminal plasma 25(OH) vitamin D level was not related to serum 25(OH) vitamin D level, but in negative relation with sperm concentration(r=0.174,P=0.016) and positive relation with semen volume(r=0.271,P=0.0001). Serum 25(OH) vitamin D level was in positive relation with seminal plasma fructose concentration(r=0.256,P=0.002), total fructose content (r=0.310,P=0.0002) and total zinc content(r=0.26,P=0.002). The level of serum and seminal plasma vitamin D leve was not related to serum anti-Mullerian hormone(AMH), seminal plasma AMH, serum inhibin (INH B) and seminal plasma INH B(P>0.05). Conclusion Vitamin D is associated with affiliated gland function. The seminal vesicles and prostate produced by semen may be the main source of vitamin D in the male reproductive system.

Knockdown of cyclin A2 expression by small interfering RNA in MG-63 cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the inhibitory effect of small interference RNA (siRNA) targeting cyclin A2 gene on the growth of osteosarcoma MG-63 and human normal skin fibroblast HSF cells and to explore whether cyclin A2 siRNAs could become a useful tool in the treatment of osteosarcoma.</p><p><b>METHODS</b>Three pairs of siRNAs targeting cyclin A2 mRNA and a pair of nonsense siRNA were designed according to the current criteria. SiRNAs were chemically synthesized and purified. The siRNAs were transfected into MG-63 cells and HSF cells via oligofectamine. The cells transfected with nonsense siRNA served as negative control group and those only treated with PBS as blank control group. Quantitative fluorescence RT-PCR, Western-blot, MTT assay, reverse transcriptase (RT)-PCR, flow cytometry and clone forming test were employed to evaluate the efficacy of RNA interference. At the same time, the mRNA expression of PCNA and cyclin B1 in siRNA-treated MG-63 cells were examined.</p><p><b>RESULTS</b>Although all three siRNAs could reduce the cyclin A2 expression, siRNA, appeared to be the most effective. After 48 h treatment with siRNA1, cyclin A2 mRNA and protein expression in MG-63 cells was significantly reduced by nearly 80% as compared with that of the blank control group, whereas the negative and blank control groups had similar expression levels. MG-63 cells treated with siRNA1 were arrested at G0/G1 phase by 80.1% and the proliferation of these tumor cells was suppressed 48 h after transfection. Furthermore, MG-63 cells showed a decreased colony forming ability after siRNA1 treatment. In addition, the cyclin A2-depleted MG-63 cells showed decreased levels of PCNA and cyclin B1. In contrast, although cyclin A2 expression in HSF reduced by nearly 60% after treatment by siRNA1 for 48h, these cells exhibited only a slight change in cell cycling, and neither clear inhibition of proliferation nor impaired colony forming ability was observed.</p><p><b>CONCLUSION</b>Cyclin A2 is critical for proliferation of MG-63 cells. Cyclin A2-siRNAs can induce obvious inhibition of cyclin A2 mRNA and protein expression in MG-63 and HSF cells, which consequently down-regulate the proliferation of MG-63 cells. There is little effect on the proliferation of siRNA-treated HSF cells. Those results indicate that siRNAs against cyclin A2 may become a potential antiproliferative tool in future antitumor therapy.</p>

Combination of small interfering RNAs mediates greater inhibition of human hepatitis B virus replication and antigen expression / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVES</b>To evaluate the inhibitory effect mediated by combination of small interfering RNAs (siRNAs) targeting different sites of hepatitis B virus (HBV) transcripts on the viral replication and antigen expression in vitro.</p><p><b>METHODS</b>(1) Seven siRNAs targeting surface (S), polymerase (P) or precore (PreC) region of HBV genome were designed and chemically synthesized. (2) HBV-producing HepG2.2.15 cells were treated with or without siRNAs for 72 h. (3) HBsAg and HBeAg in the cell culture medium were detected by enzyme-linked immunoadsorbent assay. (4) Intracellular viral DNA was quantified by real-time PCR (Polymerase Chain Reaction). (5) HBV viral mRNA was reverse transcribed and quantified by real-time PCR. (6) The change of cell cycle and apoptosis was determined by flow cytometry.</p><p><b>RESULTS</b>Our data demonstrated that synthetic small interfering RNAs (siRNAs) targeting S and PreC gene could efficiently and specifically inhibit HBV replication and antigen expression. The expression of HBsAg and HBeAg and the replication of HBV could be specifically inhibited in a dose-dependent manner by siRNAs. Furthermore, our results showed that the combination of siRNAs targeting various regions could inhibit HBV replication and antigen expression in a more efficient way than the use of single siRNA at the same final concentration. No apoptotic change was observed in the cell after siRNA treatment.</p><p><b>CONCLUSION</b>Our results demonstrated that siRNAs exerted robust and specific inhibition on HBV replication and antigen expression in a cell culture system and combination of siRNAs targeting different regions exhibited more potency.</p>

Evaluation of ST13 gene expression in colorectal cancer patients / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

We identified a novel gene ST13 from a subtractive cDNA library of normal intestinal mucosa in 1993, more studies showed that ST13 was a co-chaperone of Hsp70s. Recently we detected the ST13 gene expression in tumor tissue and adjacent normal tissue of the same colorectal cancer patient and investigated if the ST13 gene expression might have any prognostic value. Analysis was performed at molecular level by reverse transcription-PCR using real-time detection method. We measured two genes simultaneously, ST13 as the target gene and glyceraldehydes-3-phosphate dehydrogenase as a reference gene, in primary colorectal tumor specimens and tumor-adjacent normal mucosa specimens from 50 colorectal cancer patients. The expression levels of the ST13 gene were significantly decreased in primary tumors compared with adjacent mucosa (P<0.05). But there were no significant differences in the expression of ST13 as compared with different Dukes' stage, tumor differentiation grade, invasion depth, lymph node metastasis and disease-specific survival.

Studies on influence of fungal elicitor on hairy root of Panax ginseng biosynthesis ginseng saponin and biomass / 中国中药杂志

<p><b>OBJECTIVE</b>To study influence of fungal elicitors on the biomass and ginseng saponin biosynthesis of hairy roots of Panax ginseng (HRPG).</p><p><b>METHOD</b>Fungal elicitors were extracted from Colletorichum lagnarinm, Phoma filtrate, Fusarium oxysponum, Asperillus niger and culture with HRPG. The total ginseng sponin and four kinds of monomeric sponins were analysed by UV-spectrophotometry and RP-HPLC.</p><p><b>RESULT</b>Fungal elicitors coula not only can influence on HRPG biomass and total ginseng sponin, but also improve or decrease some monomeric sponin. The total ginseng sponin could be increased to 3.649% but Rg1 and Re could not be detected when A. niger elicitors wss 20 mg x L(-1) in the culture fluid.</p><p><b>CONCLUSION</b>Fungal elicitor has specificity influence on secondary metabolite of HRPG. HRPG can biosynthesize specially active component by using specific fungal elicitor is used.</p>

Measurement of the mRNA level of Polo-like kinase 1 in bronchoscopic bioptic specimens by realtime quantitative PCR / 中华检验医学杂志

0.05)and the TNM staging (P=0.55).A mild elevated compared other pathological classification was found in small cell lung cancer (0.191?0.275).Conclusions The results showed that RFQ-PCR was suitable for measurement of the mRNA level of PLKI in bronchoscopic bioptic specimens.This study suggest elevated expression of PLK1 might play a important role in development of lung cancer,so that PLK1 might be a potential tumor marker for Lung cancers.Advanced studies will be needed to clarify that PLKI mRNA level do not relate to TNM staging and pathological classification.