他莫昔芬联合人绒毛膜促性腺激素对反复种植失败患者冻融周期子宫内膜容受性的影响

目的:探讨他莫昔芬联合人绒毛膜促性腺激素(HCG)对反复种植失败(repeated implantation failure,RIF)患者冻融周期子宫内膜容受性的影响。方法:回顾性选取2017年7月至2019年6月海南省妇女儿童医学中心RIF(≥3次)再次进行冻融胚胎移植的188例患者的临床资料,根据子宫内膜准备方法不同分为观察组105例和对照组83例。观察组患者采用他莫昔芬联合HCG宫腔灌注,对照组患者采用来曲唑联合HCG宫腔灌注。比较两组患者子宫内膜厚度与内膜下血流情况[搏动指数(PI)、阻力指数(RI)]、子宫内膜血流分型、HCG日与移植日激素[黄体生成素(LH)、雌二醇(E2)及孕酮]水平及妊娠结局的差异。结果:移植日,两组患者子宫内膜厚度较促排卵前增加,且观察组患者明显大于对照组,差异均有统计学意义(P<0.05);两组患者PI、RI均较促排卵前降低,观察组患者明显低于对照组,差异均有统计学意义(P<0.05)。观察组子宫内膜血流A型患者占比为80.95%(85/105),明显高于对照组的40.96%(34/83),差异有统计学意义(P<0.05)。观察组患者HCG日与移植日的E2水平明显高于对照组,HCG日LH水平明显低于对照组,差异均有统计学意义(P<0.05)。观察组患者胚胎着床率、临床妊娠率明显高于对照组,差异均有统计学意义(P<0.05)。结论:他莫昔芬联合HCG治疗可有效改善RIF患者冻融周期子宫内膜容受性,增加子宫内膜厚度,提高妊娠率。     

保妇康栓联合干扰素栓对宫颈HPV感染患者HPV DNA负荷量、炎性因子及免疫功能的影响

目的分析保妇康栓联合干扰素栓对宫颈人乳头瘤病毒(HPV)感染患者HPV脱氧核糖核酸(HPV DNA)负荷量、炎性因子及免疫功能的影响。方法从2017年2月-2019年1月佛山市三水区人民医院收治的宫颈HPV感染患者中选取80例进行研究,以随机数字表法分为观察组和对照组,每组40例。对照组接受干扰素治疗,观察组在对照组基础上加用保妇康栓治疗,比较2组治疗效果及治疗前后HPV DNA负荷量、炎性因子及免疫功能指标。结果观察组治疗总有效率为95. 00%,高于对照组的80. 00%(P <0. 05);2组治疗前HPV DNA负荷量无明显差异(P>0. 05),观察组治疗后HPV DNA负荷量低于对照组(P <0. 05);2组治疗前高敏C-反应蛋白(hs-CRP)、降钙素原(PCT)、白介素-6(IL-6)水平无明显差异(P>0. 05),观察组治疗后炎性因子水平低于对照组(P <0. 05);2组治疗前CD3^+、CD4^+、CD8^+数值无明显差异(P> 0. 05),观察组治疗后上述免疫功能指标与对照组比较差异有统计学意义(P <0. 05)。结论保妇康栓联合干扰素栓可有效降低宫颈HPV感染患者HPV DNA负荷量,控制炎性因子水平,调节机体免疫功能,效果确切。     

Use of ex vivo and in vitro cultures of the human respiratory tract to study the tropism and host responses of highly pathogenic avian influenza A (H5N1) and other influenza viruses

The tropism of influenza viruses for the human respiratory tract is a key determinant of host-range, and consequently, of pathogenesis and transmission. Insights can be obtained from clinical and autopsy studies of human disease and relevant animal models. Ex vivo cultures of the human respiratory tract and in vitro cultures of primary human cells can provide complementary information provided they are physiologically comparable in relevant characteristics to human tissues in vivo, e.g. virus receptor distribution, state of differentiation. We review different experimental models for their physiological relevance and summarize available data using these cultures in relation to highly pathogenic avian influenza H5N1, in comparison where relevant, with other influenza viruses. Transformed continuous cell-lines often differ in important ways to the corresponding tissues in vivo.     

The influence of social evaluation on cerebral cortical activity and motor performance: A study of “Real-Life” competition

Motor performance in a social evaluative environment was examined in participants (N = 19) who completed a pistol shooting task under both performance-alone (PA) and competitive (C) conditions. Electroencephalographic (EEG), autonomic, and psychoendocrine activity were recorded in addition to kinematic measures of the aiming behavior. State anxiety, heart rate, and cortisol were modestly elevated during C and accompanied by relative desynchrony of high-alpha power, increased cortico-cortical communication between motor and non-motor regions, and degradation of the fluency of aiming trajectory, but maintenance of performance outcome (i.e., score). The findings reveal that performance in a complex social-evaluative environment characterized by competition results in elevated cortical activity beyond that essentially required for motor performance that translated as less efficient motor behavior.     

Human respiratory syncytial virus N,P and M protein interactions in HEK-293T cells

Characterization of Human Respiratory Syncytial Virus (HRSV) protein interactions with host cell components is crucial to devise antiviral strategies. Viral nucleoprotein, phosphoprotein and matrix protein genes were optimized for human codon usage and cloned into expression vectors. HEK-293T cells were transfected with these vectors, viral proteins were immunoprecipitated, and co-immunoprecipitated cellular proteins were identified through mass spectrometry. Cell proteins identified with higher confidence scores were probed in the immunoprecipitation using specific antibodies. The results indicate that nucleoprotein interacts with arginine methyl-transferase, methylosome protein and Hsp70. Phosphoprotein interacts with Hsp70 and tropomysin, and matrix with tropomysin and nucleophosmin. Additionally, we performed immunoprecipitation of these cellular proteins in cells infected with HRSV, followed by detection of co-immunoprecipitated viral proteins. The results indicate that these interactions also occur in the context of viral infection, and their potential contribution for a HRSV replication model is discussed.     

Human Sperm Anatomy: Different Expression and Localization of Phosphatidylinositol 3-Kinase in Normal and Varicocele Human Spermatozoa

Abstract

Recent reports support the possible role of PI3K in sperm capacitation and acrosome reaction, although studies regarding PI3K identity in human sperm, under certain disease states such as varicocele, are still lacking. The authors, therefore, examined the expression profile and ultrastructural localization of PI3K in human semen samples, comparing healthy donors and patients with varicocele. The results obtained performing western blotting assay showed decreased PI3K expression in varicocele with respect to the “healthy” sperm. Immunogold labeling revealed human sperm cellular compartments containing PI3K, evidencing it in the head at both the membrane and nucleus and the entire tail, from the middle to the end piece of normal sperm. In varicocele PI3K label was confined to the head, with a strong reduction of specific reaction in the neck, middle piece, and tail. In conclusion, the data suggest that PI3K may play a role in the maintenance of male factor infertility associated with varicocele, and it may be further exploited as an additional molecular marker for the diagnosis of male infertility disorders.     

Improvement of cognitive function and physical activity of aging mice by human neural stem cells over-expressing choline acetyltransferase

Aging is characterized by progressive loss of cognitive and memory functions as well as decrease in physical activities. In the present study, a human neural stem cell line (F3 NSC) over-expressing choline acetyltransferase (F3.ChAT), an enzyme responsible for acetylcholine synthesis, was generated and transplanted in the brain of 18-month-old male ICR mice. Four weeks post-transplantation, neurobehavioral functions, expression of ChAT enzyme, production of acetylcholine and neurotrophic factors, and expression of cholinergic nervous system markers in transplanted animals were investigated. F3.ChAT NSCs markedly improved both the cognitive function and physical activity of aging animals, in parallel with the elevation of brain acetylcholine level. Transplanted F3 and F3.ChAT cells were found to differentiate into neurons and astrocytes, and to produce ChAT proteins. Transplantation of the stem cells increased brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF), enhanced expression of Trk B, and restored host microtubule-associated protein 2 and cholinergic nervous system. The results demonstrate that human NSCs over-expressing ChAT improve cognitive function and physical activity of aging mice, not only by producing ACh directly but also by restoring cholinergic neuronal integrity, which might be mediated by neurotrophins BDNF and NGF.     

HTLV-1 Tax 蛋白对T细胞中HMGB1调控的影响

目的探讨人T淋巴细胞白血病1型病毒（human T-cell leukemia virus 1,HTLV-1）Tax蛋白对人高迁移率族蛋白1（ high mobility group box 1,HMGB1）基因转录调控的影响。方法提取TaxN和TaxP细胞总RNA和蛋白质,通过real-time PCR和Western blot分析HMGB1 mRNA和蛋白质的表达情况;利用脂质体介导方法,将6个含有不同长度HMGB1调控序列的pGL3-HMGB1-luc瞬时转染至TaxN 和 TaxP 细胞,观察 HMGB1基因在不同 T 细胞中的转录活性;pCMV-Tax 与 pGL3-HMGB1-luc瞬时共转染至Jurkat细胞,观察Tax蛋白对HMGB1基因的转录调控影响;染色体免疫共沉淀（ ChIP）找寻Tax蛋白影响HMGB1基因转录调控的区段。结果 TaxP细胞中HMGB1 mRNA和蛋白质表达水平高于TaxN细胞。 TaxN和TaxP细胞中HMGB1调控趋势基本相似,均表现出3号质粒（pHLuc3,含有-504～＋83 HMGB1区段）的相对荧光素酶活性（HMGB1/neo）最高,但是6号质粒（含有-1163～＋83 HMGB1区段）却表现出 TaxP 细胞 HMGB1的转录活性明显高于 TaxN 细胞。pCMV-Tax与pGL3-HMGB1-Luc报告基因共转染到Jurkat细胞也显示,6号质粒（pHLuc6）中Tax促进HMGB1基因的转录。 ChIP分析证实了Tax蛋白可能富集在HMGB1的-1163～-1043区段。结论-504～-383可能是HMGB1基因转录激活的关键启动子区,Tax蛋白可能富集在HMGB1的-1163～-1043区段促进HMGB1基因转录。     

Characterization of the human CD4+ T‐cell repertoire specific for major histocompatibility class I‐restricted antigens

While CD4⁺ T lymphocytes usually recognize antigens in the context of major histocompatibility (MHC) class II alleles, occurrence of MHC class‐I restricted CD4⁺ T cells has been reported sporadically. Taking advantage of a highly sensitive MHC tetramer‐based enrichment approach allowing detection and isolation of scarce Ag‐specific T cells, we performed a systematic comparative analysis of HLA‐A*0201‐restricted CD4⁺ and CD8⁺ T‐cell lines directed against several immunodominant viral or tumoral antigens. CD4⁺ T cells directed against every peptide‐MHC class I complexes tested were detected in all donors. These cells yielded strong cytotoxic and T helper 1 cytokine responses when incubated with HLA‐A2⁺ target cells carrying the relevant epitopes. HLA‐A2‐restricted CD4⁺ T cells were seldom expanded in immune HLA‐A2⁺ donors, suggesting that they are not usually engaged in in vivo immune responses against the corresponding peptide‐MHC class I complexes. However, these T cells expressed TCR of very high affinity and were expanded following ex vivo stimulation by relevant tumor cells. Therefore, we describe a versatile and efficient strategy for generation of MHC class‐I restricted T helper cells and high affinity TCR that could be used for adoptive T‐cell transfer‐ or TCR gene transfer‐based immunotherapies.     

Adaptive reconfiguration of the human NK‐cell compartment in response to cytomegalovirus: A different perspective of the host‐pathogen interaction

As discussed in this review, human cytomegalovirus (HCMV) infection in healthy individuals is associated with a variable and persistent increase of NK cells expressing the CD94/NKG2C activating receptor. The expansion of NKG2C⁺ NK cells reported in other infectious diseases is systematically associated with HCMV co‐infection. The functionally mature NKG2C^bright NK‐cell subset expanding in HCMV⁺ individuals displays inhibitory Ig‐like receptors (KIR and LILRB1) specific for self HLA class I, and low levels of NKp46 and NKp30 activating receptors. Such reconfiguration of the NK‐cell compartment appears particularly marked in immunocompromised patients and in children with symptomatic congenital infection, thus suggesting that its magnitude may be inversely related with the efficiency of the T‐cell‐mediated response. This effect of HCMV infection is reminiscent of the pattern of response of murine Ly49H⁺ NK cells against murine CMV (MCMV), and it has been hypothesized that a cognate interaction of the CD94/NKG2C receptor with HCMV‐infected cells may drive the expansion of the corresponding NK‐cell subset. Yet, the precise role of NKG2C⁺ cells in the control of HCMV infection, the molecular mechanisms underlying the NK‐cell compartment redistribution, as well as its putative influence in the response to other pathogens and tumors remain open issues.