Due to recurrence and resistance to chemotherapy, the current standard therapeutics are not fully effective against ovarian cancer. Therefore, we aimed to find an effective approach to improve the prognosis and therapy of ovarian cancer. NG34ScFvPD-1 is a modified oncolytic herpes simplex virus NG34 strain that expresses a single-chain antibody against programmed cell death protein 1 (PD-1) (ScFvPD-1). We assessed its efficacy and its regulatory mechanism in a mouse model of ovarian cancer. Enzyme-linked immunosorbent assay and western blot techniques were used to measure protein expression. Oncolysis caused by NG34ScFvPD-1 was examined using cytotoxicity and replication assays. The mechanism by which NG34ScFvPD-1 regulates apoptosis of ovarian cancer cells in vitro was also evaluated. We assessed the antitumor immunity and therapeutic potency of NG34ScFvPD-1 in combination with a phosphoinositide 3-kinase (PI3K) inhibitor. We found that NG34ScFvPD-1-infected ovarian cancer cells expressed and secreted ScFvPD-1, which bound mouse PD-1. The insertion of the ScFvPD-1 sequence did not inhibit the oncolytic activity of NG34ScFvPD-1, which induced apoptosis of ovarian cancer cells via the caspase-dependent pathway in vitro and activated the PI3K/AKT signaling pathway. Synergy was observed between NG34ScFvPD-1 and a PI3K inhibitor, and the combination was able to suppress tumor development, to prolong survival, and to elicit potent antitumor immunity. Thus, inhibition of PI3K enhanced the potent antitumor immunity induced by NG34ScFvPD-1 against ovarian cancer.

目的 探讨程序性细胞死亡蛋白1（PD-1）单链抗体（ScFv）溶瘤病毒对卵巢癌裸鼠模型免疫活性、肿瘤生长及存活率的影响。方法 C57BL/6小鼠右前腋下注射SKOV-3细胞构建卵巢癌裸鼠模型，分别以0、5×10~5PFU、1×10~6PFU、5×10~6PFU、1×10~7PFU、5×10~7PFU、1×10~8PFU 滴度的PD-1 Sc Fv 溶瘤病毒瘤内注射，检测小鼠肿瘤体积，筛选最佳作用滴度。构建好的卵巢癌模型裸鼠随机分为模型组、PD-1抗体组、单独溶瘤病毒组、PD-1 Sc Fv溶瘤病毒组，每组8只，再选8只小鼠右前腋下注射等量细胞培养液作为对照组。测定小鼠脾指数、胸腺指数及肿瘤体积；流式细胞实验检测小鼠外周血 T 淋巴细胞亚群 CD4 + 、CD8 + 比例与CD4 + /CD8 + 比值；以酶联免疫吸附法（ELISA）试剂盒检测小鼠血清白细胞介素（IL）-2、IL-10、γ-干扰素（IFN-γ）水平；检测小鼠外周血白细胞比例；CCK-8实验检测小鼠产生的特异性抗肿瘤细胞杀伤率；检测各组小鼠存活率。结果 与对照组比较，模型组小鼠脾指数与胸腺指数、CD4 + 比例与CD4 + /CD8 + 比值、IFN-γ与IL-2水平、外周血白细胞比例、特异性抗肿瘤细胞杀伤率、存活率降低（P＜0.05）、CD8 + 比例、IL-10水平升高（P＜0.05）；与模型组比较，PD-1 Sc Fv溶瘤病毒组、单独溶瘤病毒组、PD-1抗体组小鼠脾指数与胸腺指数、CD4 + 比例与CD4 + /CD8 + 比值、IFN-γ与IL-2水平、外周血白细胞比例、特异性抗肿瘤细胞杀伤率、存活率均升高（P＜0.05），肿瘤体积、CD8 + 比例、IL-10水平均降低（P＜0.05）；与单独溶瘤病毒组、PD-1抗体组分别比较，PD-1 Sc Fv溶瘤病毒组小鼠脾指数与胸腺指数、CD4 + 比例与CD4 + /CD8 + 比值、IFN-γ与IL-2水平、外周血白细胞比例、特异性抗肿瘤细胞杀伤率、存活率均升高（P＜0.05），肿瘤体积、CD8 + 比例、IL-10水平均降低（P＜0.05）。结论 PD-1 Sc Fv溶瘤病毒可改善卵巢癌裸鼠免疫功能，促进特异性抗肿瘤细胞产生，并增强其对癌细胞杀伤力，抑制肿瘤生长，相比单独应用PD-1抗体或溶瘤病毒疗效能更强。

目的 探讨槲皮素(Que)对紧密连接蛋白6(CLDN6)的表达及子宫内膜异位症细胞增殖、迁移和侵袭的影响,并初步分析其可能作用机制.方法 选取40例子宫内膜异位症患者为研究对象,活检时切取异位子宫内膜组织,分离与培养异位子宫内膜间质细胞(EcSCs).以不同浓度(0、12.5、25、50 μmol/L) Que干预EcSCs,通过MTT法检测不同浓度Que对EcSCs细胞增殖的影响,Western blot检测增殖细胞核抗原(PCNA)与CLDN6蛋白表达.应用Tran-swell迁移及侵袭实验检测细胞迁移及侵袭情况.EcSCs中转染阴性对照(si-control)、si-CLDN6,并分析沉默CLDN6表达对EcSCs细胞增殖、迁移和侵袭的影响.后续实验分为4组:NC组、Que组、Que+ pcDNA组、Que+ pcDNA-CLDN6组,MTT与Transwell迁移及侵袭实验分别检测各组EcSCs增殖、迁移及侵袭能力,Western blot法检测PCNA、基质金属蛋白酶-2(MMP-2)、MMP-9、上皮-间质转化(EMT)及整合素连接激酶(ILK)信号通路相关蛋白表达.结果 Que干预后EcSCs增殖、迁移及侵袭能力受到抑制,且呈剂量依赖性,可抑制CLDN6、PCNA、MMP-2、MMP-9蛋白表达;沉默CLDN6表达后能够抑制EcSCs增殖、迁移及侵袭能力;Que可抑制神经型钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、ILK、磷酸化蛋白激酶B(p-AKT)蛋白表达,促进E-cadherin蛋白表达,但转染CLDN6过表达载体后可逆转Que对EcSCs增殖、迁移及侵袭的抑制作用.结论 Que可通过抑制CLDN6表达而抑制子宫内膜异位症细胞增殖、迁移及侵袭,其可能作用机制为抑制ILK信号通路活化及EMT转换进程.

目的:探讨临床治疗原因不明复发性流产的理想方式,为原因不明复发性流产的相关实践提供借鉴和参考.方法:选取2015年1月至2016年12月期间产科收治的原因不明复发性流产患者172例,按照治疗情况把患者分成两组,观察组患者111例(进行主动免疫治疗),对照组患者61例(进行安胎治疗).结果:(1)观察组患者111例,妊娠96例,20周以前流产16例,妊娠成功80例,妊娠成功率为72.07％;对照组患者61例,妊娠50例,20周以前流产37例,妊娠成功13例,妊娠成功率为21.31％,观察组妊娠成功率高于对照组(P<0.05);(2)组间妊娠结局比较差异无统计学意义(P>0.05).结论:采用主动免疫疗法治疗原因不明复发性流产的临床效果较好.

目的 分析孕早期免疫治疗不明原因复发性流产(URSA)效果.方法 89例URSA患者,采用早期淋巴细胞免疫治疗,观察患者疗效.结果 治疗3次后封闭抗体(BA)阳性患者妊娠成功率为87.64％(78/89),显著高于治疗后阴性患者的54.55％(6/11),差异有统计学意义(P<0.05);治疗6次后阳性患者妊娠成功率为96.63％(86/89),显著高于治疗3次后妊娠成率,差异有统计学意义(P<0.05).结论 为不明原因复发性流产患者提供孕早期免疫治疗的方法,有效的提高患者妊娠率,治疗效果较好.

Objective:To investigate the effect of hypophysin with laparoscopic surgery in treatment of ovarian endometriosis and its influence on progesterone (P),luteotropic hormone (LH),estradiol (E2),follicle－stimulating hormone (FSH) and anti－Mulle－rian hormone (AMH) level.Methods:From Jul.2015 to Sep.2016,130 patients with ovarian endometriosis were selected and divided into two groups according to the different surgical methods.The observation group used hypophysin with laparoscopic surgery and the control group used laparoscopic direct ovarian cyst dissection.The operative time,intraoperative blood loss ,anal exhaust time and the level of P,LH,E2,FSH and AMH before and after operation were compared between the two groups.Results:The operative time,intra－operative blood loss and anal exhaust time in the observation group were significantly less than those in the control group (P＜0.05). One month after operation,the levels of P,LH,E2and AMH in the two groups were significantly lower than those before operation ,and the level of FSH was significantly increased (P＜0.05).Compared with the control group,the changes of P,LH,E2and FSH in the ob－servation group were significantly less than those in the control group (P＜0.05).Three months after operation,the levels of P,LH,E2 and FSH became normal in the observation group ,but still lower in the control group compared with those before the operation (P＜0.05),the AMH level of the observation group was significantly higher than that of the control group ( P＜0.05).Conclusions:Hy－pophysin with laparoscopic surgery in treatment of ovarian endometriosis can shorten the operation time ,reduce intra－operative blood loss,significantly reduce the changes of P ,LH,E2and FSH levels,effectively reserve ovarian function and improve the quality of life.

Objective To investigate the efficacy and safety of combination of different drug on adolescent polycystic ovary syndrome (PCOS).Methods Sixty adolescent patients with polycystic ovary syndrome from January 2015 to January 2017 were randomly divided into group A,B and C,with 20 ca-ses in each group.Patients in group A were given estrogen (Diane-35 )treatment,patients in group B were given metformin plus Diane-35 treatment,patients in group C were given combined regimen (Diane-35 +metformin and pioglitazone)treatment.The clinical efficacy,adverse reactions,fasting in-sulin (FIN)and fasting blood sugar (FBS)levels and serum sex hormones were compared cross the three groups.Results The total effective rate in group A,B,C was 75. 00%,60. 00%,95. 00%,respec-tively,there was a significant difference among the three groups (P＜0. 05 );though the total incidence of adverse reactions in group A and B was higher than that in group C,there was no significant difference among the three groups (P＞0. 05 ).After treatment,levels of FIN and FBS of patients in the three groups all were improved,and group C got most remarkable improvement,and there were significant differences in FIN and FBS level among the three groups (all P＜0. 05 ).After treatment,the plasma sex hormone (testosterone,luteinizing hormone,follicle-producing hormone,luteinizing hormone/follicle-producing hormone)levels in group C were all decreased,and were lower than those of another two groups (P＜0. 05 ).Conclusions Diane 35 ,metformin and pioglitazone combination method has excel-lent clinical curative effect and certain degree of security.At the same time,it can improve insulin resist-ance and androgen levels in patients,which is worth promoting.

目的 研究促卵泡生成激素受体基因多态性与卵巢过度刺激综合征的相关性,为临床防治提供参考依据.方法 选取2015年1月-2016年12月期间在我院接受体外受精(卵胞浆内单精子显微注射ICSI)治疗的120例不孕症患者作为研究对象,所有患者均在就诊当天采集血液标本,并对血液样本实施FSHR基因680位点、307位点分型鉴定.结果 本研究120例患者中出现28例OHSS,在OHSS组、非OHSS组中患者的680位点基因型GA、AA、GG与等位基因频率A、G分布比较,差异有统计学意义(P＜0.05);在OHSS组、非OHSS组中患者的307位点基因型GA、AA、GG与等位基因频率A、G分布比较,差异无统计学意义(P＞0.05).结论 卵巢过度刺激综合征发生可能与促卵泡生成激素受体基因的Asn680Ser位点有关,而关于Thr307Ala位点与卵巢过度刺激综合征发生的相关性有待进一步研究.

Objective:To study the relationship between the gene polymorphism of methylene tetrahydrofolate reductase (MTHFR) and the unexplained recurrent spontaneous abortion (URSA).Methods:83 patients with unexplained recurrent spontaneous abortion (the URSA group) and 90 normal women of reproductive age (the control group) were recruited.Fluorescence quantitative PCR was used to detect the MTHFR gene C677T,A1298C single nucleotide polymorphisms (SNP).The frequency distribution of genotype and allele was compared,and the risk of different genotypes to URSA was evaluated.Results:The distribution of MTHFR genotypes in both groups was consistent with the Hardy-Weinberg balance.There were significant differences in the frequencies of C677T genotype and alleles between the URSA group and the control group (P＜0.05).There were no significant differences in the frequencies of A1298C genotype and alleles between the two groups (P＞0.05).The frequency of homozygous genotype in the URSA group was higher than that in the control group (30.1％ vs.13.3％,x2=7.237,P=0.007).The risk of URSA in those patients with TT homozygous genotype was 3.289 times higher than that in the patients with CC homozygous genotype (OR=3.289,95％CI:1.342-8.063).Conclusions:The gene polymorphism of MTHFR C677T site is a risk factor of URSA,and the TT homozygoous genotype increases the risk of URSA.

目的 分析血浆D-二聚体水平与血小板聚集功能在反复自然流产患者中的诊断价值.方法 102例反复自然流产患者为观察组,对其血浆D-二聚体水平与血小板聚集功能进行检测,另选取100例健康妇女(无自然流产史、非孕期)为对照组,对两组进行对比.结果 观察组患者血浆D-二聚体中位值为0.27 mg/L,对照组中位值为0.15 mg/L,比较差异具有统计学意义(P<0.05).两组二磷酸腺苷(ADP)诱导血小板凝集率异常比较差异无统计学意义(P>0.05),D-二聚体水平异常比较差异具有统计学意义(P<0.01).结论 反复自然流产患者血小板处于高凝状态,对血浆D-二聚体水平与血小板聚集功能进行检测,具有临床诊断价值,为疾病的进一步治疗提供一定参考价值.

目的 探讨反复自然流产(recurrent spontaneous abortion,RSA)患者血清中孕激素诱导阻断因子(PIBF)及Th1/Th2型细胞因子的异常表达及其临床意义.方法 采用酶联免疫吸附法检测94例RSA患者血清中PIBF和Th1(IL-2、TNF-α、IFN-γ)/Th2(IL-10)型细胞因子水平,同时以72例正常未孕妇女及92例正常健康早孕妇女作为对照.结果 3组研究对象外周血血清中各细胞因子的表达水平组间比较均有显著性差异,RSA组血清中IL-2、TNF-α、IFN-γ的水平显著高于正常健康早孕组,差异有统计学意义(P<0.05),而PIBF、IL-10水平显著低于正常健康早孕组,差异有统计学意义(P<0.05).结论 RSA患者血清中PIBF表达水平降低可导致反复自然流产,可作为预测反复自然流产的检测指标;Th1/Th2型细胞因子间平衡的改变与反复自然流产有关,而PIBF在调节Th1/Th2型细胞因子的平衡中起重要的作用.

Objective:To evaluate the possible mechanisms of pathogenesis of severe intrauterine adhesions ( IUA) ,the effects of oxidized regenerated cellulose ( ORC) in treatment of severe IUA and expression of transforming growth factor-β1 ( TGF-β1 ) . Methods:60 pa-tients with severe IUA were randomly divided into the observation group (30 patients) and the control group (30 patients) . After operation,intrauterine device ( IUD) and ORC were inserted into uterine cavity in the observation group,while the control group used IUD only. All cases underwent postoperative artificial cycles for successive 3 months and the second-look hysteros-copy. The expression of TGF-β1 in the fibrosis tissue and the normal endometrium were mesured by immunohistochemistry method. Results:Before operation,the expression of TGF-β1 in the fibrosis tissue was significantly higher than that in the normal endometrium ( P<0 . 01 ) . In each groups,the postoperative expression of TGF-β1 was significantly higher than before op-eration in the normal endometrium and also in the fibrosis tissue (P<0. 01). After operation, TGF-β1 of the observation group was significantly lower than that of the control group in the fi-brosis tissue ( P<0 . 01 ) . The total effective rate of the observation group was significantly higher than that of the control group(P<0. 05). In re-adhesion cases,the degree of adhesion was sig-nificantly lower in observation group than that in control group ( P<0 . 05 ) . Conclusions:The abnomal expression of TGF-β1 may be relavant to the occurrence of severe IUA,ORC could im-prove the effective rate,downgrade the degrees of re-adhesion and decrease expression of TGF-β1.