多囊卵巢综合征患者血清microRNA-27a、microRNA-320的表达及其临床意义

目的探讨血清microRNA-27a(miR-27a)、microRNA-320(miR-320)在多囊卵巢综合征(PCOS)患者中的表达及临床意义。方法选取2017年1月—2020年8月海南西部中心医院收治的180例PCOS患者,根据胰岛素抵抗指数(HOMA-IR)分为IR组113例(HOMA-IR≥2.58)和NIR组67例(HOMAIR<2.58);另选取该院体检的健康女性80例作为对照组。于月经第3~5天采集静脉血,检测血清miR-27a、miR-320的表达,以及性激素、糖脂代谢、胰岛素抵抗指标。经阴道超声测量卵巢体积并计数卵泡数。分析miR-27a、miR-320表达与BMI、性激素、糖脂代谢、胰岛素抵抗、卵巢体积、卵泡数的相关性及PCOS发病的影响因素。结果IR组BMI、血清睾酮(T)、总胆固醇(TC)、胰岛素抵抗(HOMA-IR)、miR-27a mRNA的相对表达量高于NIR组和对照组(P<0.05),miR-320 mRNA的相对表达量低于NIR组和对照组(P<0.05)。NIR组BMI、血清T、TC、HOMA-IR、miR-27a mRNA的相对表达量高于对照组(P<0.05),miR-320 mRNA的相对表达量低于对照组(P<0.05)。血清miR-27a的表达与BMI、卵泡数、卵巢体积、T、TC、HOMA-IR呈正相关(r=0.749、0.835、0.764、0.867、0.907和0.882,均P<0.05),血清miR-320表达与卵泡数、卵巢体积、T、HOMA-IR呈负相关(r=-0.791、-0.808、-0.768和-0.752,均P<0.05)。卵泡数增加[OR=1.865(95%CI:1.252,2.776)]、T水平升高[OR=1.536(95%CI:1.179,2.001)]、HOMA-IR增加[OR=1.702(95%CI:1.185,2.446)]、miR-27a高表达[OR=1.933(95%CI:1.278,2.923)]、miR-320低表达[OR=0.610(95%CI:0.467,0.796)]是PCOS易感因素(P<0.05)。联合miR-27a、miR-320诊断PCOS的AUC为0.917(95%CI:0.869,0.964),高于单独诊断的0.708(95%CI:0.637,0.779)和0.676(95%CI:0.602,0.750)。结论PCOS患者血清miR-27a表达上调,miR-320表达下调,miR-27a高表达、miR-320低表达与PCOS易感和胰岛素抵抗有关,可作为PCOS诊断指标。

Clinical study of different fertilization methods of assisted reproductive technology on neonatal birth defects

Objective:To investigate the incidence of neonatal birth defects in assisted reproductive technology(ART)by in vitro fertilization(IVF)and intracytoplasmic sperm injection(ICSI).Methods:The clinical data of 4229 cases of singleton deliver by infertile patients under 35 years old who received IVF/ICSI-ET in our center were analyzed.According to different fertilization methods,they were divided into IVF group(2967 cases)and ICSI group(1262 cases).The general situation of birth,birth defects and the location of defects were compared between the two groups.Results:a total of 38 cases of neonatal birth defects were found,the incidence of birth defects was 0.89%,including 30 cases(1.01%)in IVF group and 8 cases(0.64%)in ICSI group.There was no significant difference in the incidence of birth defects between the two groups(P>0.05).There was also no significant difference in birth weight,gestational age and gender ratio between the two groups(P>0.05).Conclusion:Different fertilization methods in assisted reproductive technology do not increase the incidence of neonatal birth defects.

Effects of clenbuterol exposure on PPARγ expression in adipocytes of rats

Objective To investigate effects of clenbuterol （CLB） on the peroxisome proliferators- activated receptor γ （PPARγ） expression in adipose tissues of rats. Methods Twenty adult female Sprague-Dawley rats were randomly divided into 4 groups （5 rats per group）. CLB solved in normal saline solution was given at the dose of 0 mg/kg body weight （bw） （group A, as the control）, 0.4 mg/kg bw （group B, low-dose group）, 2.0 mg/kg bw （group C, mid-dose group）, and 18.5 mg/kg bw （group D, high-dose group）for 14 d by gavage consecutively, respectively. Methods of immunohistochemistry, quantitative Real-time PCR and Western blotting were performed to detect expression of PPARγ in the adipose tissue samples. Results PPARγ-positive immunostaining was strong in the controls and weak in the experimental groups. There was no difference on PPARγmRNA and protein between the low-dose group and the control （P〉0.05）. With the increase of CLB doses, expression levels of PPARγmRNA and protein were significantly lower in mid- or high-dose group than those in the control （19〈0.01）. Conclusions The PPARγ expression in adipose tissues of rats could be down-regulated after CLB exposure, and the decrease became more severe with the increasing doses.

Hemospermia is not associated with the production of antisperm antibody

Objective To investigate whether hemospermia is involved in the production of antisperm antibodies （ASA）. Methods A total of 40 males with hemospermia were enrolled in this study. These males were divided into 3 groups according to their fertile status： 18 cases were patients who visited the infertility clinic （group A）, 8 had fertility （group B）, and 14 did not know their fertile status （group C）. Semen samples were collected and ASA levels were detected by the direct immunobead test （D-IBT）. Leukocytes in semen were assessed by the peroxi- dase assay. Results None of these males with hemospermia were found to display significant levels of ASA in the 3 groups. The number of cases who had increased levels of leukocytes in the semen samples of groups A, B and C were 15 （83.3%）, 6 （75.0%） and 12 （85.7%）, respectively. Conclusion Hemospermia is not associated with the production of ASA. It is worth noting that the leukocyte levels are increased in semen of males with hemospermia.

Effects of Different Serum on in vitro Cultured Human Granulosa Cells

Objective To evaluate the effects of different sera on the growth of human granulosa cells （GCs） cultured in vitro. Methods GCs were obtained from women who underwent follicular aspirates during in vitro fertilization （IVF） program. Five groups were divided according to media supple- mented with different types of sera. Group A, 10% heat-inactivated fetal bovine serum （FBS）; group B, 10% heat-inactivated newborn bovine serum; group C, 10% non heat-inactivated FBS; group D, 10% human serum; group E, bovine serum albumin. Morphological characteristics and viability of GCs measured by trypan blue exclusion assay were evaluated after 24 h of incubation.Results GCs cultured in group A and group B showed multiform morphology compared with other groups. GCs cultured in group D and group E were present with cytoplasmic atrophy and less pseudopodium. Moreover, group A and group B showed a similar level in the viability of GCs （P〉0.05）, which displayed no difference between group D and group E as well （P〉0.05）. Group C had a lower level of viability than group A （P〈0.05） but a higher level than group D （P〈0.01）. Conclusion Heat-inactivated sera can improve the growth of GCs. Different types of sera would have different effects on the growth of GCs cultured in vitro. The pre-culture with different types of sera should be performed to get better efficacy.