Chromosome polymorphisms and their influence on semen quality and sperm DNA integrity in males undergoing IVF/ICSI / 中华男科学杂志

Objective@#To investigate the incidence of chromosome polymorphisms and their influence on semen quality and sperm DNA integrity in male patients receiving in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI).@*METHODS@#We retrospectively analyzed the chromosomal karyotypes and the types and incidence rate of chromosome polymorphisms in 2 370 male patients undergoing IVF/ICSI between June 2016 and June 2018. We classified the patients into groups A (with variation in the secondary constriction region in the autosomal long arm), B (with variation in the short arm of the D/G group chromosomes), C (with interbrachial inversion of chromosome 9) and D (with Y chromosome polymorphisms), and compared the semen parameters and sperm DNA fragmentation indexes (DFI) between the patients with chromosome polymorphisms and those with normal chromosomes.@*RESULTS@#Totally, 154 (6.50%) of the patients undergoing IVF/ICSI were found with chromosome polymorphisms, including 34 cases of secondary constriction variation in the long arm of the autosome (1.43% ［34/2 370］, 22.08% ［34/154］), 82 cases of short arm polymorphisms of the D/G group chromosomes (3.46% ［82/2 370］, 53.25% ［82/154］), 26 cases of interbrachial inversion of chromosome 9 (1.10% ［26/2 370］, 16.88% ［26/154］), 10 cases of Y chromosome polymorphisms (0.42% ［10/2 370］, 6.50% ［10/154］), and 2 cases of mixed chromosome polymorphisms (0.08% ［2/2 370］, 1.42% ［2/154］). The total sperm count was lower in group D than in the other polymorphism groups and the normal chromosome group, but with no statistically significant difference among the five groups (P > 0.05). The sperm progressive motility was also lower in group D than in the other five groups, with statistically significant difference from group B (27.5 ± 13.5 vs. 41.5 ± 21.1, P = 0.027), but not from the other groups (P > 0.05). No statistically significant difference was observed in the sperm DFI between the polymorphism groups and the normal chromosome group (P > 0.05), or among the polymorphism groups (P > 0.05). The proportion of normal semen was lower in group D than in the other four groups, but with no statistically significant difference among the five groups (P > 0.05). The incidence rate of asthenospermia was higher in group D than in the other four groups, but with no statistically significant difference among the five groups (P > 0.05), and so was that of oligoasthenospermia, with statistically significant difference from the normal chromosome group (30.0% vs 8.0%, P = 0.041), but not from the other polymorphism groups (P > 0.05).@*CONCLUSIONS@#Short arm polymorphisms of the D/G group chromosomes are the most common type of chromosome polymorphisms in male patients undergoing IVF/ICSI. Polymorphisms of the Y chromosome have a negative effect on semen quality, while those of the other chromosomes do not significantly affect semen quality and sperm DNA integrity.

Liuwei Dihuang Decoction improves cognitive impairments via regulating immune system in APP/PS1 transgenic mouse,a mouse model of Alzheimer disease / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effect and mechanisms of Liuwei Dihuang Decoction (LW)on cognition in PrP-hAβPPswe/PS1ΔE9(APP/PS1)transgenic mice.METHODS LW was adminis-trated with oral for 3 months.The locomotor activity test was performed to investigate the spontaneous motor activity of mice. The Morris water maze test and shuttle box test were performed to investigate the spatial learning and memory and active avoidance response respectively.The Αβ deposits and neuron loss in the hippocampus was detected by immunofluorescence staining and nissl staining respectively. The flow cytometry was employed to investigate the lymphocyte subsets of the mice.The 3H-thymidine incorporation was performed to investigate the splenocytes proliferation. RESULTS The treatment of LW ameliorated the impairments of spatial learning and memory and active and passive avoidance in APP/PS1 mice. The administration of LW alleviated neuron loss in the brain, suppressed amyloid-β (Αβ) deposits in the hippocampus of APP/PS1 mice. The treatment of LW significantly increased ConA-and LPS-induced proliferation of splenocytes,increased CD3+T cells and CD19+B cells in the spleen lymphocytes and reduced Gr1+cells in APP/PS1 mice.CONCLUSION This data indicated the adminis-tration of LW ameliorated behavioral and pathological deterioration via regulating immune function.

LW-AFC,a new formula derived from Liuwei Dihuang decoction,ameliorates behavioral and pathological deterioration via modulating the neuroendocrine-immune system in Alzheimer disease mouse models / 中国药理学与毒理学杂志

OBJECTIVE Alzheimer disease(AD),the most common cause of dementia among older people, could not be prevented, halted, or reversed up till now. A large body of pharmacological study has revealed that Liuwei Dihuang (LW) possesses potential therapeutic effects on AD. LW-AFC is key fractions from LW.In the present study,we investigated the effect of LW-AFC on AD mouse models. METHODS PrP-hAβPPswe/PS1ΔE9(APP/PS1) mice and senescence-accelerated mouse prone 8 strain (SAMP8), classic AD animal models, were employed. After the treatment of LW-AFC, mice were cognitively evaluated in behavioral experiments. Neuron loss, amyloid-β (Αβ) deposition, and Αβ level were analyzed using Nissl staining, immunofluorescence, and an AlphaLISA assay, respectively. Multiplex bead analysis, a radioimmunoassay, immunochemiluminometry, and an ELISA were used to measure cytokine and hormone levels.Lymphocyte subsets were detected using fl ow cytometry. RESULTS LW-AFC ameliorated the cognitive impairment observed in APP/PS1and SAMP8 mice,including the impairment of object recognition memory,spatial learning and memory,and active and passive avoidance. In addition, LW-AFC alleviated the neuron loss in the hippocampus, suppressed amyloid-β(Αβ)deposition in the brain,and reduced the concentration of Aβ1-42in the hippo-campus and plasma of APP/PS1 mice. LW-AFC treatment also significantly restored the imbalance of hypothalamic-pituitary-adrenal (HPA) and hypothalamic-pituitary-gonadal (HPG) axis, enhanced the proliferation of splenocytes,corrected the disorder of lymphocyte subsets,and regulated the abnormal production of cytokine in APP/PS1 and SAMP8 mice. Effects of LW-AFC on pharmacodynamics and neuroendocrine immunomodulation network in APP/PS1 and SAMP8 mice were better than meman-tine and donepezil. CONCLUSION LW-AFC ameliorated the behavioral and pathological deterioration of AD mouse models via the restoration of the NIM network, which supports the use of LW-AFC as a potential agent for AD therapy.

Normal sperm morphology and the outcomes of routine in vitro fertilization / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the application value of morphology assessment of sperm from fresh semen in routine in vitro fertilization (IVF).</p><p><b>METHODS</b>We analyzed the morphology of the sperm from fresh or optimized semen samples and, based on the sperm morphology of the raw semen, allocated 908 IVF cycles due to the pure tubal factor to different groups: morphologically normal sperm (MNS) ≤ 4%, > 4% - ≤ 15%, and > 15% in Trial 1 and MNS ≤ 1%, > 1% - ≤ 2%, > 2% - ≤ 3%, and > 3%-- ≤ 4% in Trial 2. We compared the rates of fertilization, cleavage, high-quality embryo, -blastocyst formation, and pregnancy among different groups.</p><p><b>RESULTS</b>The total fertilization rate was significantly lower in the MNS ≤ 4% than in the MNS > 4% - ≤ 15% and >15% groups (74.40% vs 78.61% and 80.03%, P < 0.01). Compared with the MNS ≤ 1%, > 1% - ≤ 2%, and > 2% - ≤ 3% groups, the MNS > 3% - ≤ 4% group showed remarkably increased rates of 2PN normal fertilization (77.23%, 78.97% and 78.99% vs 85.47%, P < 0.01), cleavage (95.71%, 96.01% and 97.27% vs 98.73%, P < 0.05), and blastocyst formation (53.85%, 49.01% and 49.55% vs 63.41%, P < 0.01). No statistically significant differences were observed in the rates of clinical pregnancy, implantation, early abortion, live birth, or malformation at birth among different groups (P > 0.05).</p><p><b>CONCLUSION</b>MNS ≤ 4% affected the total rate of fertilization while MNS ≤ 3% reduced the rate of normal fertilization in IVF. However, even MNS ≤ 1% did not result in fertilization disorder or failure. Therefore, teratozoospermia alone was not an indicator of ICSI and sperm mor- phology assessment had no obvious value for predicting the rates of embryo quality, clinical pregnancy, and live birth in IVF.</p>

Anti-sense miRNA-21 oligonucleotide inhibits Tb 3.1 human tongue squamous cell carcinoma growth in vitro / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of micro RNA-21 (miRNA-21) knocking on the Tb3.1 human tongue squamous cell carcinoma growth.</p><p><b>METHODS</b>Anti-sense miRNA-21 oligonucleotide was delivered with oligofectamine to suppress Tb 3.1 tongue cancer cell growth in vitro. Real-time polymerase chain reaction (PCR) was conducted to detect the miRNA-21 expression after transfection. Methyl thiazolyl tetrazolium (MTT) assay was used to determine Tb 3.1 cell survival rate. Apoptosis were examined by flow-cytometry. Matrigel matrix and transwell assay were used to determine Tb 3.1 cell colony formation and migration ability. Antigen KI-67 (Ki67), B cell lymphoma (Bcl-2), phosphatase and tensin homolog (PTEN), matrirx metalloproteinase 2 (MMP-2, MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) protein expression in Tb 3.1 cell were measured by Western blotting.</p><p><b>RESULTS</b>miRNA-21 expression was decreased in miRNA-21 antisense oligonucleotide (ASODN) group. The survival rate of Tb 3.1 cells with AS-miRNA-21 transfection was significantly suppressed (F = 27.02, P = 0.00) and early phase apoptosis (F = 26.641, P = 0.001) induced in Tb 3.1 cell. Ki67, Bcl-2, MMP-2 and MMP-9 protein were down regulated while PTEN and TIMP-1 protein expression was increased.</p><p><b>CONCLUSIONS</b>Blocking miRNA-21 expression in Tb3.1 cell could suppress cancer cell growth in vitro and miRNA-21 can serve as a novel target candidate for human tongue cancer gene therapy.</p>

Study on effect and mechanism of liuwei dihuang decoction in modulating hypothalamus-pituitary-ovary axis in senescence accelerated mice model / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the changes of hypothalamus-pituitary-ovary (HPO) axis of senescence accelerated mice (SAM) model in their aging process and the modulatory effect of Liuwei Dihuang decoction (LW) on the changes.</p><p><b>METHODS</b>Using vaginal smear to test the estrous cycle of animal; using radioimmunoassay to determine serum levels of estradiol (E2), and beta-endorphin (beta-EP) and substance P (SP) in hypothalamus; and the western bloting technique was adopted to quantify the level of luteinizing hormone (LH) in pituitary and estradiol receptor alpha (ER alpha) in pituitary and ovary.</p><p><b>RESULTS</b>The estrous cycle and diestrus were significantly prolonged as SAMP8 aged, the concentration of serum E2 was lower, pituitary LH was higher in the SAMP8 than those in SAMR1. Content of beta-Ep and SP in hypothalamus gradually lowered during aging, SP content lowered after a transient increasing, and ER alpha expression lowered significantly. LW could significantly shorten the diestrus, increase weight of ovary, elevate serum E2 level, significantly reduce the LH level in pituitary, increase beta-EP content in hypothalamus and ER alpha expression in ovary, but significantly lower SP content in hypothalamus. Oral administration of estrogen could increase serum E2, beta-EP and SP content in hypothalamus and ER alpha expression in pituitary, but lower LH level in pituitary, weight of ovary and ER alpha level in ovary.</p><p><b>CONCLUSION</b>SAMP8, in the aging process, develops a progressive HPO axis functional disturbance, it is closely related with the change of levels of peptide neuro-mediator in hypothalamus and change of ER alpha level in ovary. LW could improve or modulate the HPO axis functional disturbance in SAMP8.</p>

Effect of Liuwei Dihuang decoction on secretion of testosterone in the testis leydig cells of senescence accelerated mouse / 军事医学科学院院刊

Objective:To study the effect of Liuwei Dihuang decoction (LW) on secretion of testosterone in the testis leydig cells of senescence accelerated mouse (SAM). Methods:The level of testosterone in the testis leydig cells of SAM with aging and the effect of LW on the secretion of testosterone were observed using cultured testis leydig cells in vitro.Results:The level of testosterone in the testis leydig cells of SAMP8 with aging was significantly decreased and showed a significant difference compared with age-matched SAMR1. Chronic administration of LW (2.5 and 5.0 g/kg) for 5 months significantly ameliorated the secretion of testosterone in SAMP8 compared with control group. Conclusions:The secretory function of testis leydig cells was reduced in SAMP8 with aging. LW ameliorated the secretion of testosterone in the testis leydig cells,indicating that LW could antagonize or delay the deterioration of the testis leydig cells in SAMP8.