Correlation analysis of vitamin D level and anti-Müllerian hormone in infertile female and the role in predicting pregnancy outcome / 北京大学学报(医学版)

OBJECTIVE@#To investigate the relationship between serum 25(OH)D and anti-Müllerian hormone (AMH) among infertile females and their predictive impacts on in vitro fertilization and embryo transfer pregnancy outcome.@*METHODS@#Totally 756 infertile females treated with assisted reproductive technology were enrolled and divided into three groups according to their vitamin D levels (group A with serum 25(OH)D≤10 μg/L, group B with serum (10-20) μg/L, and group C with serum ≥20 μg/L). The serum AMH levels were detected. The differences among the groups were analyzed, as well as the correlation between vitamin D levels and serum AMH levels in various infertility types (fallopian tube/male factor, polycystic ovary syndrome (PCOS), ovulation disorders excluded PCOS, endometriosis, unexplained infertility, and others). Also, the predictive roles of vitamin D and AMH in pregnancy outcome in all the infertile females were discussed.@*RESULTS@#(1) 87.7% of the enrolled females were insufficient or deficient in vitamin D. (2) The serum AMH levels in the three groups with different vitamin D levels were 1.960 (1.155, 3.655) μg/L, 2.455 (1.370, 4.403) μg/L, 2.360 (1.430, 4.780) μg/L and there was no significant difference in serum AMH levels among the three groups (P>0.05). (3) Serum 25(OH)D and AMH levels presented seasonal variations (P < 0.05). (4) There was no prominent correlation between the serum AMH level and serum 25(OH)D level in females of various infertility types after adjusting potential confounding factors [age, body mass index (BMI), antral follicle count (AFC), vitamin D blood collection season, etc.] by multiple linear regression analysis (P>0.05). (5) After adjusting for confounding factors, such as age, BMI, number of transplanted embryos and AFC, the results of binary Logistics regression model showed that in all the infertile females, the serum AMH level was an independent predictor of biochemical pregnancy outcome (P < 0.05) while the serum 25(OH)D level might not act as a prediction factor alone (P>0.05). In the meanwhile, the serum 25(OH)D level and serum AMH level were synergistic predictors of biochemical or clinical pregnancy outcome (P < 0.05).@*CONCLUSION@#Based on the current diagnostic criteria, most infertile females had vitamin D insufficiency or deficiency, but there was not significant correlation between serum 25(OH)D and ovarian reserve. While vitamin D could not be used as an independent predictor of pregnancy outcome in infertile females, the serum AMH level could predict biochemical pregnancy outcome independently or jointly with vitamin D.

Identification of risk genes in Chinese nonobstructive azoospermia patients based on whole-exome sequencing / 亚洲男科学杂志(英文版)

Nonobstructive azoospermia (NOA) is a severe condition in infertile men, and increasing numbers of causative genes have been identified during the last few decades. Although certain causative genes can explain the presence of NOA in some patients, a proportion of NOA patients remain to be addressed. This study aimed to investigate potential high-risk genes associated with spermatogenesis in idiopathic NOA patients by whole-exome sequencing. Whole-exome sequencing was performed in 46 male patients diagnosed with NOA. First, screening was performed for 119 genes known to be related to male infertility. Next, further screening was performed to determine potential high-risk causative genes for NOA by comparisons with 68 healthy male controls. Finally, risk genes with high/specific expression in the testes were selected and their expression fluctuations during spermatogenesis were graphed. The frequency of cystic fibrosis transmembrane conductance regulator (CFTR) gene pathogenic variant carriers was higher in the NOA patients compared with the healthy controls. Potential risk genes that may be causes of NOA were identified, including seven genes that were highly/specifically expressed in the testes. Four risk genes previously reported to be involved in spermatogenesis (MutS homolog 5 [MSH5], cilia- and flagella-associated protein 54 [CFAP54], MAP7 domain containing 3 [MAP7D3], and coiled-coil domain containing 33 [CCDC33]) and three novel risk genes (coiled-coil domain containing 168 [CCDC168], chromosome 16 open reading frame 96 [C16orf96], and serine protease 48 [PRSS48]) were identified to be highly or specifically expressed in the testes and significantly different in the 46 NOA patients compared with 68 healthy controls. This study on clinical NOA patients provides further evidence for the four previously reported risk genes. The present findings pave the way for further functional investigations and provide candidate risk genes for genetic diagnosis of NOA.

Cryobiological Characteristics of L-proline in Mammalian Oocyte Cryopreservation / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>L-proline is a natural, nontoxic cryoprotectant that helps cells and tissues to tolerate freezing in a variety of plants and animals. The use of L-proline in mammalian oocyte cryopreservation is rare. In this study, we explored the cryobiological characteristics of L-proline and evaluated its protective effect in mouse oocyte cryopreservation.</p><p><b>METHODS</b>The freezing property of L-proline was detected by Raman spectroscopy and osmometer. Mature oocytes obtained from 8-week-old B6D2F1 mice were vitrified in a solution consisting various concentration of L-proline with a reduced proportion of dimethyl sulfoxide (DMSO) and ethylene glycol (EG), comparing with the control group (15% DMSO and 15% EG without L-proline). The survival rate, 5-methylcytosine (5-mC) expression, fertilization rate, two-cell rate, and blastocyst rate in vitro were assessed by immunofluorescence and in vitro fertilization. Data were analyzed by Chi-square test.</p><p><b>RESULTS</b>L-proline can penetrate the oocyte membrane within 1 min. The osmotic pressure of 2.00 mol/L L-proline mixture is similar to that of the control group. The survival rate of the postthawed oocyte in 2.00 mol/L L-proline combining 7.5% DMSO and 10% EG is significantly higher than that of the control group. There is no difference of 5-mC expression between the L-proline combination groups and control. The fertilization rate, two-cell rate, and blastocyst rate in vitro from oocyte vitrified in 2.00 mol/L L-proline combining 7.5% DMSO and 10% EG solution are similar to that of control.</p><p><b>CONCLUSIONS</b>It indicated that an appropriate concentration of L-proline can improve the cryopreservation efficiency of mouse oocytes with low concentrations of DMSO and EG, which may be applicable to human oocyte vitrification.</p>

Female fertility: is it safe to "freeze?" / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To evaluate the safety and risk of cryopreservation in female fertility preservation.</p><p><b>DATA SOURCES</b>The data analyzed in this review were the English articles from 1980 to 2013 from journal databases, primarily PubMed and Google scholar. The criteria used in the literature search show as following: (1) human; embryo; cryopreservation/freezing/vitrification, (2) human; oocyte/immature oocyte; cryopreservation/ freezing/vitrification, (3) human; ovarian tissue transplantation; cryopreservation/freezing/vitrification, (4) human; aneuploidy/DNA damage/epigenetic; cryopreservation/freezing/vitrification, and (5) human; fertility preservation; maternal age.</p><p><b>STUDY SELECTION</b>The risk ratios based on survival rate, maturation rate, fertilization rate, cleavage rate, implantation rate, pregnancy rate, and clinical risk rate were acquired from relevant meta-analysis studies. These studies included randomized controlled trials or studies with one of the primary outcome measures covering cryopreservation of human mature oocytes, embryos, and ovarian tissues within the last 7 years (from 2006 to 2013, since the pregnancy rates of oocyte vitrification were significantly increased due to the improved techniques). The data involving immature oocyte cryopreservation obtained from individual studies was also reviewed by the authors.</p><p><b>RESULTS</b>Vitrifications of mature oocytes and embryos obtained better clinical outcomes and did not increase the risks of DNA damage, spindle configuration, embryonic aneuploidy, and genomic imprinting as compared with fresh and slow-freezing procedures, respectively.</p><p><b>CONCLUSIONS</b>Both embryo and oocyte vitrifications are safe applications in female fertility preservation.</p>

Combination of serum inhibin B and follicle-stimulating hormone levels can not improve the diagnostic accuracy on testicular sperm extraction outcomes in Chinese non-obstructive azoospermic men / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>It is still controversial whether the serum inhibin B level is a superior predictor of the presence of sperm in testicular sperm extraction (TESE) in azoospermic men compared with serum follicle-stimulating hormone (FSH). In this study, we evaluated the diagnostic accuracy of serum inhibin B levels as a predictor of the outcome of TESE in Chinese non-obstructive azoospermic men and compared it with the traditional marker serum FSH and testicular volumes.</p><p><b>METHODS</b>Basal values of serum hormone levels, testicular volumes and histological evaluation of 305 Chinese non-obstructive azoospermic men were analyzed. The level of inhibin B was measured using a three-step enzyme-linked immunoassay before sperm extraction, and the diagnostic accuracy of prediction of the outcome of TESE was compared for different markers by the receiver operating characteristics (ROC) curve analysis.</p><p><b>RESULTS</b>Testicular sperm was successfully retrieved in 137 of 305 patients (44.9%). The serum level of inhibin B, the FSH and the testicular volume were significantly different between the successful TESE group and the unsuccessful group. According to the ROC curve analysis, for inhibin B, the cut-off value for discriminating between successful and failed TESE was 28.39 pg/ml (sensitivity 83.5%, specificity 79.1%). For FSH, the best cut-off value for discriminating was 11.05 pg/ml (sensitivity 83.5%, specificity 74.5%). The area under the ROC curve of serum inhibin B was similar to that of FSH. Combining the serum inhibin B with FSH levels did not improve the predictive value for successful TESE.</p><p><b>CONCLUSIONS</b>Serum inhibin B and FSH levels are correlated with spermatogenesis. However, inhibin B is not superior to FSH in predicting the presence of sperm in TESE. And the combination of them does not improve the diagnostic accuracy on TESE outcome.</p>

Effect of curettage and copper wire on rabbit endometrium: a novel rabbit model of endometrial mechanical injury / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>It remains almost a helpless situation for the recurrent implantation failure and pregnancy loss caused by endometrial injury at present. The purpose of this study was to develop a rabbit model of endometrial mechanical injury that could provide a research platform for this difficult clinical predicament.</p><p><b>METHODS</b>Three experiments were conducted. Experiment 1: Curettages in both uterus horns and copper wire inserting after curettage (double-injury) in one horn. The histological changes were monitored at 0, 24, 48, 72 hours, as well as in 1 and 2 weeks after operation. Experiment 2: Direct copper wire inserting in one horn and double-injury in other horn. The wires in both horns were removed after 2 weeks. The histological changes were recorded at 0, 1 and 2 weeks after wire removal. Experiment 3: Double-injury procedure in one horn was performed and wire was removed after 2 weeks; another horn was remained normal to serve as control. Histological changes were recorded, tissue areas were measured, and proliferation indices (PIs, %) were calculated at 1, 2, 4 and 8 weeks after wire removal, respectively.</p><p><b>RESULTS</b>The experiments revealed that the injured endometrium by simple curettage or copper wire could be fully repaired. While the endometrial regeneration was severely impaired by double-injury, both areas of endometrium and uterine cavity decreased (P < 0.05); both PIs of glandular epithelial and stromal cells increased and reached maximum at 4 weeks (P < 0.05), but returned by 8 weeks.</p><p><b>CONCLUSION</b>This study demonstrated that a rabbit model of endometrial injury could be effectively established through a double-injury procedure of curettage and copper wire with comparable clinical index.</p>

Inhibitory effect of lentiviral vectors expressing small interfering RNA targeting survivin gene on Hep-2 cell growth in vitro and in nude mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of the lentiviral vectors expressing small interfering RNA (siRNA) for survivin gene knockdown in inhibiting Hep-2 cell growth in vitro and its tumorigenicity in nude mice.</p><p><b>METHODS</b>The tumorigenicity of Hep-2 cells transfected with the siRNA mediated by the lentiviral vectors was tested in nude mice. The expression of survivin gene of the transfected cells at the mRNA and protein levels were detected by RT-PCR and Western blotting, respectively, and the cell cycle changes were analyzed by flow cytometry.</p><p><b>RESULTS</b>Transfection of the siRNA targeting survivin significantly decreased the expression of survivin mRNA and protein in Hep-2 cells in vitro by 60%-85% and 70%, respectively, resulting also in increased cell apoptosis as shown by flow cytometry (P<0.01). The transfection significantly lowered the tumorigenicity of the cells in nude mice.</p><p><b>CONCLUSION</b>The lentiviral vectors expressing survivin siRNA can significantly inhibit survivin gene expression in Hep-2 cells and induce the cell apoptosis in vitro, and suppress the tumorigenicity of the cells in nude mice.</p>

Reversal effect of resveratrol on chemotherapy resistance in KBv200 cell line and underlying mechanisms / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To explore the reversal effect and potential mechanism of resveratrol on multidrug resistance of human oral epidermoid carcinoma KBv200 cells.</p><p><b>METHODS</b>MTT assay was used to investigate reversal index of resveratrol to vincristine, adriamycin and paclitaxel. Cell apoptosis were measured by flow cytometry. RT-PCR and Western blot were used to detect mRNA and protein expression of multidrug resistant 1 (MDR1) and B cell lymphoma leukemia-2 (Bcl-2).</p><p><b>RESULTS</b>Resveratrol produced a synergistic effect with chemotherapeutics and obviously reversed the multidrug resistant phenotype of KBv200 cells. The reversal fold (RF) of 200 micromol/L resveratrol to vincristine, paclitaxel and adriamycin were 77.1, 61.3 and 5.9, respectively. The gene array results showed that resveratrol greatly downregulated expression levels of Bcl-2 and MDR1. After treated with 100 micromol/L, 200 micromol/L resveratrol, the expression level of Bcl-2 and MDR1 in KBv200 cells were markedly decreased in comparison with those untreated (t were 2.98, 3.51 and 3.12, 4.56, P < 0.05).</p><p><b>CONCLUSIONS</b>Resveratrol can efficiently reverse multidrug resistance in KBv200 cells. The potential mechanism may be via inhibiting the multidrug resistant gene expressions and/or promoting cell apoptosis.</p>

Protective effect of adeno-associated virus-mediated neurotrophin-3 on the cochlea of guinea pigs with gentamicin-induced hearing loss / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the protective effect of local gene therapy with adeno-associated virus (AAV)-mediated neurotrophin-3 (NT-3) on the function and morphology of the cochlea of guinea pigs with gentamicin-induced hearing loss.</p><p><b>METHODS</b>Hearing loss was induced with gentamicin (80 mg.kg(-1).day(-1) injected intramuscularly) in 18 pigmented guinea pigs 4 days prior to gene transfer. The guinea pigs were then divided into groups A, B, and C for AAV-mediated NT-3 gene transfer (n=7), AAV infection (n=7) or no particular intervention (n=4), respectively. Mini-Osmotic pump were implanted in either side of the ears in groups A and B, and the guinea pigs were injected with gentamicin (80 mg.kg(-1).day(-1)) intramuscularly since the operation day for 10 consecutive days. In group C, only gentamicin was administrated. Before and 14 days after gentamicin administration, auditory brainstem response audiometry (ABR) and distort-product otoacoustic emissions (DPOAE) were recorded, and the animals sacrificed to observe the morphological changes of the cochlear microscopically.</p><p><b>RESULTS</b>Compared with groups B and C, the animals in group A showed better auditory ability (ABR and DPOAE) and significantly higher surviving rate of the outer hair cells (P<0.05).</p><p><b>CONCLUSION</b>AAV-mediated NT-3 gene transfer may protect and repair the cochlear hair cells and auditory function damaged by aminoglycoside ototoxicity in guinea pigs, and aseptic procedure is of vital importance in cochlear local gene therapy.</p>

Surgical treatment of hyperthyroidism / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>In order to explore the management of peri-operation and the therapeutic effect in the surgical treatment of hyperthyroidism.</p><p><b>METHODS</b>Fifty five cases of hyperthyroidism were undergone near-total thyroidectomy, during the operation recurrent laryngeal nerve was exposed, and the parathyroid was found with microscope when necessary. The third rank of inferior thyroid arteries were ligated to guarantee the blood supply for parathyroid.</p><p><b>RESULTS</b>All cases underwent near-total thyroidectomy. There was no mortality, and no permanent recurrent laryngeal nerve palsy occurred, and no permanent hypoparathyroidism, and no recurrent hyperthyroidism. Follow-up was carried out 16 months to approximately 5 years after near-total thyroidectomy patients, Hypothyroidism occurred in 15 cases (57.7%), serum calcium levels were 2.15-2.45 mmol/L.</p><p><b>CONCLUSIONS</b>Special attention should be given to the management of peri-operation, the above the method can prevent operative complication in the surgical treatment of hyperthyroidism, with excellent result.</p>

Polymorphism of methionine synthase gene in nuclear families of congenital heart disease / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the relation of methionine synthase (MS) gene variation with congenital heart disease (CHD) phenotype.</p><p><b>METHODS</b>One hundred and ninety three CHD patients (94 males and 99 females) and their biological parents (nuclear families) in Liaoning Province were selected as the case group, and another 104 normal persons (60 males and 44 females) and their parents without family history of birth defects as the control group. For all subjects the polymorphism of MS gene A2756G locus was examined by PCR-RFLP method.</p><p><b>RESULTS</b>In offspring of the control group the frequencies of MS genotype (+/-) and allele (+) were 10.7% and 5.3%, without existence of homozygote. The MS genotype distribution and allele frequencies of CHD patients and their mothers were not significantly different from the control (P > 0.05). The frequency of allele (+) in case fathers (5.0%) was apparently lower than that in the control (9.1%, P = 0.060), and the odds ratio (OR) was 0.53 (95% CI: 0.25-1.09). There was no difference in parents' genotype combination between the two groups, and in genotype distribution among different types of CHD. Analysis of genetic transmission indicated that mutation allele (+) existed transmission disequilibrium in CHD nuclear families. The percentage of allele (+) transmitted from parents was lower than that allele (-) with OR 0.26 (95% CI: 0.11-0.60).</p><p><b>CONCLUSION</b>MS gene variation in parents is associated with occurrence of CHD in offspring, and mutation allele (+) in parents may be related with the decrease of CHD risk in offspring.</p>