目的  探讨精子DNA碎片指数（DFI）对体外受精-胚胎移植（IVF-ET）胚胎发育及妊娠结局的影响，为优化男性生育力评估及辅助生殖治疗策略提供依据。方法  回顾性分析2023年1月—2024年1月于徐州市妇幼保健院接受IVF-ET治疗的126对不孕夫妇，根据男方DFI检测结果分为低碎片组（DFI≤15%，n=42）、临界组（15%＜DFI＜30%，n=45）和高碎片组（DFI≥30%，n=39）。比较三组患者受精相关指标、胚胎发育指标及妊娠结局指标的差异，并分析DFI与各指标的相关性。结果  低碎片组双原核率（2PN）率、优质胚胎率及囊胚形成率均高于临界组和高碎片组（P＜0.001），低碎片组1PN率、多PN率均低于临界组和高碎片组（P＜0.001）；三组种植率、临床妊娠率、早期流产率比较差异无统计学意义（P＞0.05），但高碎片组活产率低于低碎片组（P＜0.05）。相关性分析结果表明，DFI与优质胚胎率（r=-0.412，P＜0.001）、囊胚形成率（r=-0.387，P＜0.001）、活产率（r=-0.287，P=0.012）呈负相关，与早期流产率（r=0.206，P=0.059）、种植率（r=-0.215，P=0.058）、临床妊娠率（r=-0.203，P=0.072）无显著相关性。结论  精子DNA碎片指数是影响IVF-ET胚胎发育及妊娠结局的重要因素，高DFI主要通过降低胚胎发育潜能及增加流产风险导致活产率下降，临床需对高DFI患者进行干预以改善治疗结局。

       Objective  To investigate the impact of sperm DNA fragmentation index（DFI）on embryo development and pregnancy outcomes of in vitro fertilization-embryo transfer（IVF-ET），and to provide a basis for optimizing male fertility assessment and assisted reproductive treatment strategies．Methods  A retrospective analysis was performed on 126 infertile couples undergoing IVF-ET treatment at the Reproductive Medicine Center of Xuzhou Maternal and Child Health Hospital from January 2023 to January 2024．According to the male DFI test results，they were divided into three groups：low fragmentation group（DFI≤15%，n=42），critical group（15% ＜ DFI ＜ 30%，n=45），and high fragmentation group（DFI≥30%，n=39）．Differences in fertilization-related indicators，embryo development indicators，and pregnancy outcome indicators were compared among the three groups，and the correlation between DFI and each indicator was analyzed．Results  The 2 pronuclei rate（PN）rate，high-quality embryo rate，and blastocyst formation rate in the low fragmentation group were significantly higher than those in the critical and high fragmentation groups（P＜0.001）．The 1PN  rate and multi-PN  rate in the low fragmentation group were significantly lower than those in the critical and high fragmentation groups（P＜0.001）．There was no significant difference in the three groups of implantation rate，clinical pregnancy rate and early abortion rate（P＞0.05），but the live birth rate of high fragment group was significantly lower than that of low fragment group（P＜0.05）．The results of correlation analysis showed that DFI was significantly negatively correlated with the rate of high quality embryos（r=-0.412，P＜0.001），blastocyst formation rate（r=-0.387，P＜0.001）and live birth rate（r=-0.287，P=0.012），but not with the rate of early abortion（r=0.206，P=0.059），implantation rate（r=-0.215，P=0.058）and clinical pregnancy rate（r=-0.203，P=0.072）．Conclusions  Sperm DFI is an important factor affecting embryo development and pregnancy maintenance in IVF-ET．High DFI leads to a decrease in live birth rate mainly by reducing embryo developmental potential and increasing the risk of early abortion．Clinically，early intervention is needed for patients with high DFI to improve treatment outcomes．

目的 探讨鳞状上皮细胞抗原（SCCA）、人乳头瘤病毒（HPV）-DNA联合阴道镜检查在宫颈鳞状细胞癌（SCC）筛查中的应用价值。方法 选择2019年1月1日—2021年12月31日在中山市博爱医院就诊并确诊为SCC的妇女作为研究对象,共纳入100例SCC患者（SCC组）,同时选择200例经活检确诊为宫颈慢性炎症的患者（宫颈慢性炎症组）作为阴性对照。采用阴道镜观察研究对象的宫颈情况,并采集研究对象的宫颈组织标本进行HPV-DNA检测。采集研究对象的静脉血,采用化学发光免疫法测定研究对象SCCA的水平。以病理检查结果为金标准,分别对HPV-DNA检测、外周血SCCA两者联用以及阴道镜、HPV-DNA检测、外周血SCCA三者联用进行筛查效果的评价。结果 SCC组研究对象的年龄≥40岁者、出血者、性生活开始年龄≤20岁者比例均高于宫颈慢性炎症患者组;而宫颈慢性炎症患者组疼痛的比例高于SCC患者组,差异均有统计学意义（P<0.01）。χ²检验结果显示,SCC组研究对象的SCCA阳性率高于宫颈慢性炎症组（P<0.001）。阴道镜结合SCCA、HPV-DNA检测筛查SCC的灵敏度和特异度均高于单独使用SCCA和HPV-DNA,并取得较好的约登系数（75%）和Kappa值（0.730）。结论 采用阴道镜结合HPV-DNA、SCCA可有效提高SCC疾病的约登系数与Kappa值,其联合诊断的效能高于单独使用阴道镜、HPV-DNA或SCCA诊断SCC。

Objective To study the application value of squamus cell carcinoma antigen（SCCA）and human papillomavirus（HPV）-DNA combined with colposcope in cervical squamous cell carcinoma（SCC）screening．Methods Women diagnosed with SCC who visited Boai Hospital of Zhongshan city from January 1,2019 to December 31,2021 were selected as research subjects,including 100 patients with SCC（SCC group）and 200 patients with chronic cervical inflammation confirmed by biopsy（chronic cervical inflammation group）．The cervix of the subjects was observed by colposcope,and cervical tissue samples were collected for HPV-DNA testing．Venous blood of subjects was collected and SCCA levels were determined by chemiluminescence immunoassay．Using pathological examination results as the gold standard,the screening effect of combination HPV-DNA test and peripheral blood SCCA test,combination colposcope,HPV-DNA test and peripheral blood SCCA were evaluated respectively．Results In SCC group,the proportion of age≥40 years old,bleeding,sexual life age ≤20 years old were higher than those in chronic cervical inflammation group,but chronic cervical inflammation group had higher rate of pain than those in SCC group（P<0.01）by Chi-square test．SCCA positive rate in SCC group was higher than that in chronic cervical inflammation group（P<0.001）by Chi-square test．The sensitivity and specificity of colposcope combined with SCCA and HPV-DNA were higher than those of SCCA and HPV-DNA alone,and better Youden’s coefficient（75%）and Kappa value（0.730）were obtained．Conclusions Colposcope combined with HPV-DNA and SCCA can effectively improve the Youden’s coefficient and Kappa value of SCC disease,and its combined diagnosis efficiency was higher than that of colposcope,HPV-DNA and SCCA alone in the diagnosis of SCC,which has high clinical promotion significance．

目的 使用TCGA数据库,探索DNA甲基化位点对肺腺癌的预后影响。方法 使用TCGA数据库,获取肺腺癌病人癌和癌旁组织甲基化表达数据、基因表达数据及临床数据;将人群分为探索组和验证组,使用LASSO在探索人群中筛选对肺腺癌预后有影响的甲基化位点;受试者工作特征曲线用于评估甲基化位点预测效果,并进一步在验证人群中验证。结果 在TCGA数据库中筛选出158个癌和癌旁组织差异表达且与所在基因mRNA表达显著相关的甲基化位点,经LASSO回归分析,cg19378330与肺腺癌预后相关。甲基化位点水平高于中位数的患者,归入高风险组,甲基化位点水平低于中位数的为低风险组。结果发现与低风险组相比,高风险组的死亡风险比低风险组增加了38%(OR=1.38,95% CI=1.16～2.69)。在探索阶段人群中其曲线下面积为0.80(95% CI=0.73～0.88),灵敏度为86.2%。验证人群中也表现出类似结果。结论 甲基化位点cg19378330与肺腺癌具有较显著的关联性,且可以对肺腺癌的风险进行有效的预测。

Objective Using the TCGA database to explore the prognostic effects of DNA methylation sites on lung adenocarcinoma. Methods TCGA database was used to collecting DNA methylation data, gene expression data and clinical data of lung adenocarcinoma patients. The population were divided into the exploratory group and the validation group. The LASSO regression analysis was used to screen the methylation sites associated with the prognosis of lung adenocarcinoma in the exploratory group. Receiver operating characteristic curve was used to evaluate the prediction effect of the model, and further verified in the validation population. Results A total of 158 methylation sites with differential expression and significant correlation with the mRNA expression of the corresponding gene were screened from the TCGA database. With LASSO regression analysis, the DNA methylation sites associated with prognosis of lung adenocarcinoma were cg19378330. Those patients with levels above the median methylation site were assigned to the high-risk group, while those with levels below the median methylation site were assigned to the low-risk group. Patients' death risk in the high-risk group were 38% higher than those in the low-risk group (OR=1.38, 95%CI=1.16-2.69). The area under the curve was 0.80 (95%CI=0.73-0.88) and the sensitivity was 86.2% in the exploratory stage population.Similar results were seen in the validation population. Conclusions The DNA methylation site cg19378330 was significantly associated withthe prognosisof lung adenocarcinoma, and could effectively predict the risk of lung adenocarcinoma.

目的 探讨p16免疫细胞化学、人乳头瘤病毒(HPV)DNA基因分型单独和联合检测在宫颈细胞学不能明确诊断意义的非典型鳞状上皮细胞(ASC-US)分流中的价值。方法 收集2017年3月—2022年1月,585例液基薄层细胞学(TCT)诊断为ASC-US患者的宫颈细胞学标本,使用免疫细胞化学法行p16蛋白检测,生物芯片法行HPV DNA基因分型检测,患者于8周内行阴道镜下病理活检术。以组织学诊断结果为金标准,探讨p16免疫细胞化学和HPV DNA基因分型单独和联合检测方法在同一级别宫颈病变中阳性率的差异,对比同一检测方法在不同级别宫颈病变中阳性率的差异,比较p16免疫细胞化学、HPV DNA基因分型单独和联合检测对高级别鳞状上皮内病变(HSIL)以上病变诊断效能的差异,综合评定一种最优的ASC-US分流方法。结果 ①(p16+HPV DNA)联合检测在同一级别宫颈病变中阳性率高于p16免疫细胞化学、HPV DNA基因分型检测。②p16免疫细胞化学、HPV DNA基因分型、(p16+HPV DNA)联合检测的阳性率均随着宫颈病变程度的加重而递增。③(p16+HPV DNA)联合检测的综合诊断效能最佳,其灵敏度、特异度、符合率和约登指数分别为99.07%、62.55%、69.23%、0.62。结论 p16免疫细胞化学检测法与HPV DNA基因分型单独和联合检测均有助于ASC-US分流,但是,(p16+HPV DNA)联合检测具有更优的灵敏度和约登指数,同时保持了较高的特异度和符合率,可有效进行ASC-US分流。

Objective To investigate the value of p16 immunocytochemistry and human papillomavirus (HPV) DNA genotyping alone and combined in atypical squamous cells of undetermined significance (ASC-US) shunt which cervical cytology can not clearly diagnose. Methods From March 2017 to January 2022, cervical cytological specimens of 585 patients with ASC-US diagnosed by liquid based thinprep cytology test (TCT) were collected. p16 protein was detected by immunocytochemistry, HPV DNA genotype was detected by biochip and the patients underwent colposcopy pathological biopsy within 8 weeks. Taking the histological diagnosis results as the gold standard, the differences of the positive rate of p16 immunocytochemistry and HPV DNA genotyping in the same level of cervical lesions, differences of the positive rate of the same detection method in different levels of cervical lesions and differenes of p16 immunocytochemistry HPV DNA genotyping alone and combined detection of the diagnostic efficacy of lesions severer than high-grade squamous intraepithelial lesion (HSIL) were compared to comprehensively evaluate an optimal ASC-US shunt method. Results ①The positive rate of combined detection of (p16+HPV DNA) in the same level of cervical lesions was higher than that of differences of p16 immunocytochemistry and HPV DNA genotyping. ②The positive rate of combined detection of (p16+HPV DNA), p16 immunocytochemistry and HPV DNA genotyping increased with the aggravation of cervical lesions. ③The combined detection of (p16+HPV DNA) had the best comprehensive diagnostic efficiency and its sensitivity, specificity, coincidence rate and Yoden index were 99.07%, 62.55%, 69.23% and 0.62 respectively. Conclusions p16 immunocytochemical assay and HPV DNA genotyping, both alone and in combination, contributed to ASC-US shunt. However, the combined detection of (p16+HPV DNA) had better sensitivity and Yoden index, with high specificity and coincidence rate, which can effectively carry out ASC-US shunt.

目的 通过研究统计痰TB-DNA、痰分枝杆菌核酸、痰涂片找抗酸杆菌、血清T-SPOT.TB试验对肺结核的诊断敏感度、特异度、诊断预测值、诊断准确率,进一步探讨不同临床检测方法对肺结核的诊断价值,指导肺结核患者的临床诊治。方法 通过回顾性分析我院2017年1月—2019年12月呼吸内科、感染性疾病科诊断为活动性肺结核的患者,以痰结核菌培养结果为对照标准,分别统计出痰TB-DNA、痰分枝杆菌核酸、痰涂片找抗酸杆菌、血T-SPOT.TB试验对肺结核的诊断敏感度、特异度、阳性预测值、阴性预测值、诊断准确率,探讨我院临床上四种实验室方法对诊断肺结核的临床价值。结果 通过上述方法统计出痰TB-DNA、痰分枝杆菌核酸、痰涂片找抗酸杆菌、血T-SPOT.TB试验对肺结核的诊断敏感度分别是84.7%、88.1%、74.7%、96.0%,特异度分别是65.3%、69.2%、86.5%、17.8%,阳性预测值分别是83.0%、85.%、92.0%、70.7%,阴性预测值分别是68.1%、73.5%、62.1%、68.4%,诊断准确率分别是78.2%、82.0%、78.5%、70.5%。结论 跟传统方法痰结核菌培养、痰涂片找抗酸杆菌比较,TB-DNA、分枝杆菌核酸、TB-SPOT.TB试验在时效、灵敏度方面更具优势,能敏感检测出人体是否感染肺结核,对患者的早期诊断及指导治疗具有重要意义。

Objective To investigate the diagnostic sensitivity, specificity, predictive value and diagnostic accuracy of TB-DNA, mycobacterium sputum nucleic acid, acid-fast bacilli on sputum smear and serum T-SPOT.TB test for tuberculosis, so as to further explore the significance of different clinical detection methods for tuberculosis and guide the clinical diagnosis and treatment of tuberculosis patients. Methods By retrospective analysis of January 2017-December 2019, patients from respiratory medicine, infectious diseases departments diagnosed with active tuberculosis, sputum culture results of tuberculosis bacterium as control standard, we figured out sputum TB-DNA, sputum mycobacterium nucleic acid blood, sputum smear for acid fast bacilli, T-SPOT. TB test to the diagnosis sensitivity, specific degree, positive predictive value, negative predictive value, diagnostic accuracy, to explore the clinical value of four clinical laboratory methods in our hospital. Results According to the above methods, the diagnostic sensitivity of sputum TB-DNA, sputum mycobacterial nucleic acid, sputum acid-fast bacilli on smear and blood T-SPOT.TB test for tuberculosis was 84.7%、88.1%、74.7%、96.0%, and the specificity was 65.3%、69.2%、86.5%、17.8%, respectively. The positive predictive value was 83.0%、86.6%、92.0%、70.7%, and the negative predictive value was 68.1%、73.5%、62.1%、68.4%, respectively. The diagnostic accuracy was 78.2%、82.0%、78.5%、70.5%, respectively. Conclusion Compared with the traditional methods of culture and sputum smear for acid-fast bacilli, TB-DNA, mycobacterial nucleic acid and T-SPOT.TB test had more advantages in terms of timeliness and sensitivity. It is great significance for the early diagnosis and treatment of patients to detect whether they are infected with tuberculosis sensitively.

目的 我们探讨2019年6月—2020年1月复发性流产夫妇男性患者精浆弹性蛋白酶同精液参数及DNA碎片率的可能关系。方法 研究对象纳入80例复发性流产的男性患者及25例因女方输卵管因素行IVF-ET正常生育的男性患者。精液标本用来进行精浆弹性蛋白酶、精液常规分析、精子核染色质分析及精子形态学等参数分析。结果 结果表明同正常生育男性相比,复发性流产的弹性蛋白酶是增高(P=0.010)。我们将复发性流产男性患者分为正常组(<600 ng/mL)及异常组(≥600 ng/mL)。结果表明异常组患者的精子前向运动比例(P=0.002)及正常形态百分率(P=0.009)均降低,而精子DNA碎片率(P=0.002)增高。Spearman相关性分析发现精浆弹性蛋白酶同精子前向运动比例(r=-0.43,P<0.001)及正常形态百分率(r=-0.39,P<0.001)负相关,而同精子DNA碎片率(r=0.36,P=0.001)正相关。结论 精浆弹性蛋白酶可能影响复发性流产男性患者的精子活力、形态及DNA碎片率。复发性流产男性患者的生殖道隐性感染值得重视,其相关临床探讨性值得深入研究。

Objective Our study is aim to investigate the possible relationship of seminal elastase, on semen parameters and DNA fragmentation in male patients of recurrent pregnancy loss (RPL) between June 2019 and January 2020. Methods The patients included 80 male patients of RPL couple and 25 male patients from couples with clinical pregnancy through in vitro fertilization due to the female tubal factor. The semen samples were used to determine the seminal elastase, computed assisted semen analysis, sperm dispersion test and sperm morphology analysis. Results Compared to the control group, the levels of seminal elastase was increased in the RPL group. The RPL group was divided into the normal group (Elastase<600 ng/mL) and abnormal group (Elastase ≤ 600 ng/mL).The abnormal group exhibited the lower percentage of progressive sperm (P=0.002) and normal morphology (P=0.009),but higher precentage of DNA fragmentation (P=0.002). Meanwhile, the seminal elastase was positively associated with DNA fragmentation (r=0.36,P=0.001), but was inversely associated with the sperm motility (r=-0.43,P<0.001) and normal morphology (r=-0.39,P<0.001). Conclusion Our study may unveil the possible effects of the seminal elastase on the semen parameters and DNA fragmentation in the male patients of RPL couples. Further studies should put more emphasis on the silent genital tract inflammation of the patients.

目的 探讨一次不明原因流产与精子DNA损伤的关系。方法 收集有一次不明原因流产史的患者作为实验组,同时以有正常妊娠史的患者为对照组,分别比较两组男方年龄、精子密度、精子活力、精液量和精子DNA断裂指数有无差异。以SPSS 16.0为统计软件,进行独立样本的t检验。结果 两组的精液量、精子密度及活力均无差异,实验组男方年龄小于对照组(P<0.05),但实验组的DFI要高于对照组(P<0.01)。结论 本研究对照组年龄高于实验组,而DFI正好相反。这说明不明原因的自发流产与男方精子DFI密切相关,随着DFI的增加,流产风险增加。

Objective To investigate the relationship between unexplained abortion and sperm DNA damage. Methods Patients with a history of unexplained abortion were enrolled as an experimental group, and patients with a normal pregnancy history were used as a control group. The differences in age, sperm density, sperm motility, semen volume, and sperm DNA break index were compared between the two groups. The independent sample t test was performed using the statistical software of SPSS 16.0. Results There was no significant difference in semen volume, sperm density and motility between the two groups. The age of the male group in the experimental group was lower than that in the control group (P<0.05), but the DFI of the experimental group was higher than that of the control group, and the difference was statistical significance (P<0.01). Conclusion The age of the study control group was higher than that of the experimental group, while DNA fragmentation index DFI was the opposite, which indicated that unexplained spontaneous abortion was closely related to the male sperm DFI, and the risk of miscarriage may increase with the increase of DFI.

目的 观察不同剂量卡介苗核酸(Bacille Calmette-guerin DNA , BCG-DNA)在不同干预时间对哮喘小鼠气道高反应性及气道炎症的干预作用。方法 1.将Balb/c雌鼠随机分为哮喘模型组、NS对照组、BCG- DNA干预组。干预组根据干预的时间和干预制剂剂量的不同分为-7DNA1 μg、-7DNA10 μg、-7DNA100 μg、10DNA1 μg、10DNA10 μg、10DNA100 μg、17DNA1 μg、17DNA10 μg、17DNA100 μg组。2.在末次激发48小时后,测定各浓度级乙酰甲胆碱激发下的增强的呼气间歇 (Enhanced Pause, Penh)值,将其与小鼠激发前吸入NS后的Penh的百分比(Penh%NS),作为其气道反应性评价指标;其次对肺泡灌洗液进行细胞学分析。结果 1.气道反应性:①-7DNA1 μg组从Mch为3.12～50 mg/mL之间的Penh%NS显著低于哮喘组(P<0.05);②-7DNA10 μg组和-7DNA 100 μg组从Mch为6.25～25 mg/mL之间的Penh%NS显著低于哮喘组(P<0.05);③10DNA10 μg组从Mch为12.5～25 mg/mL之间的Penh%NS显著低于哮喘组(P<0.05);④10DNA100 μg组从Mch为3.12、12.5～50 mg/mL之间的Penh%NS显著低于哮喘组(P<0.05);⑤17DNA1 μg组在Mch为3.12、12.5 mg/mL的Penh%NS显著低于哮喘组(P<0.05);⑥17DNA10 μg组在Mch为12.5 mg/mL之间的Penh%NS显著低于哮喘组(P<0.05) 2.气道炎症:10DNA1 μg、-7DNA10 μg、10DNA10 μg和17DNA10 μg组的BALF细胞分类Eos%分别为:35.34±3.81、27.30±6.91、38.20±6.56、42.17±5.17;显著低于哮喘组的Eos%(48.8±6.12)(P<0.05);10DNA1 μg组的Eos%显著低于-7DNA1 μg组的Eos%(P<0.05);-7DNA10 μg组的Eos%显著低于10DNA10 μg、17DNA10 μg、-7DNA1 μg和-7DNA100 μg组的Eos%(P<0.05)。结论 BCG-DNA能降低哮喘小鼠的气道高反应性,减轻哮喘小鼠的气道炎症,早期(－7 d)中小剂量的干预效果较佳。

Objective To investigate the effect of Bacille Calmette-Guerin BCG-DNA on airway hyperresponsiveness and airway inflammation in asthmatic mouse model. Methods 1.According to different intervention, mouse were divided into asthma groups, NS control group, BCG-DNA group. According to different time and dosage intervened with asthma model, the BCG-DNA group were subdivided into -7DNA1 μg、-7DNA10 μg、-7DNA100 μg、10DNA1 μg、10DNA10 μg、10DNA100 μg、17DNA1 μg、17DNA10 μg and 17DNA100 μg group. 2.48 hours after the final incitation, the mice were stimulated with increasing concentrations of methacholine, and the airway resistance was measured. Enhance pause (Penh) was taken for each group. Bronchoalveolar lavage cytology was performed to evaluate the airway inflammation. Results 1.Airway hyperresponsiveness: ① Penh%NS of-7DNA1 μg group was significantly lower than the asthma group when Mch was 3.12~50 mg/mL (P<0.05); ② Penh%NS of -7DNA10 μg group and -7DNA100 μg group were significantly lower than the asthma group when Mch was 6.25~50 mg/mL (P<0.05);③ Penh%NS of 10DNA10 μg group was significantly lower than the asthma group when Mch was 12.5~25 mg/mL (P<0.05); ④ Penh%NS of 10DNA100 μg group was significantly lower than the asthma group when Mch was 3.12,12.5~50 mg/mL (P<0.05); ⑤ Penh%NS of 17DNA1 μg group was significantly lower than the asthma group when Mch was 3.12 or 12.5 mg/mL (P<0.05);⑥Penh%NS of 17DNA10 μg group was significantly lower than the asthma group when Mch was 12.5 mg/mL(P<0.05). 2.Airway inflammation: The Eos% of 10DNA1 μg, -7DNA10 μg,10DNA10 μg and 17DNA10 μg group (35.34±3.81、27.30±6.91、38.20±6.56、42.17±5.17) were lower than the asthma group (P<0.05); The Eos% of 10DNA1 μg group was lower than the -7DNA1 μg group (P<0.05); The Eos% of -7DNA10 μg group was lower than the 10DNA10μg, 17DNA10 μg,-7DNA1 μg and -7DNA100 μg group (P<0.05). Conclusion BCG-DNA can inhibit the airway inflammation and hyperresponsiveness in asthmatic mouse model. Early interventions with middle dose bring better results.