益肾化瘀方辅助治疗对不明原因复发性流产患者辅助性T细胞17/调节性T细胞平衡及雌孕激素的影响

目的观察益肾化瘀方辅助治疗对不明原因复发性流产(URSA)患者辅助性T细胞17(Th17)/调节性T细胞(Treg)平衡及雌孕激素的影响。方法将92例URSA患者按照随机数字表法分为2组,对照组46例予西医常规治疗,治疗46例在对照组基础上联合益肾化瘀方治疗,连续治疗至孕12周。比较2组中医证候积分、Th17/Treg细胞水平、Th17和Treg相关细胞因子[白细胞介素17A(IL-17A)、白细胞介素35(IL-35)、转化生长因子β_(1)(TGF-β_(1))]、雌孕激素[血清雌二醇(E_(2))、孕酮(P)、β人绒毛膜促性腺激素(β-HCG)]水平、中医证候疗效、保胎成功率。结果治疗组总有效率96.65%(44/46),对照组总有效率82.61%(38/46),治疗组高于对照组(P<0.05)。2组治疗后中医证候各项评分及总分均较本组治疗前降低(P<0.05),且治疗组治疗后均低于对照组(P<0.05)。2组治疗后Th17细胞、Th17/Treg均较本组治疗前降低(P<0.05),Treg细胞均升高(P<0.05),且治疗组治疗后Th17细胞、Th17/Treg均低于对照组(P<0.05),Treg细胞高于对照组(P<0.05)。2组治疗后血清IL-17A均较本组治疗前降低(P<0.05),IL-35、TGF-β_(1)均升高(P<0.05),且治疗组治疗后血清IL-17A水平低于对照组(P<0.05),IL-35、TGF-β_(1)水平均高于对照组(P<0.05)。2组治疗后血清E_(2)、P、β-HCG均较本组治疗前升高(P<0.05),且治疗组治疗后E_(2)、P、β-HCG均高于对照组(P<0.05)。治疗组保胎成功率84.78%(39/46),对照组65.22%(30/46),治疗组高于对照组(P<0.05)。结论益肾化瘀方辅助治疗能够改善URSA临床症状,提高保胎成功率,可能与纠正Th17/Treg平衡失调、促进雌孕激素分泌等因素有关。

高危型人乳头瘤病毒分型和液基薄层细胞学检查在宫颈癌前病变和宫颈癌筛查中的应用价值

目的:探讨高危型人乳头瘤病毒(high-risk human papillomavirus,HR-HPV)分型检测与液基薄层细胞学检查(thin-prep cytology test,TCT)在宫颈癌前病变及宫颈癌筛查中的临床价值。方法:选取2020年4月至2021年3月在南京市中西医结合医院妇科门诊进行宫颈癌筛查的150例患者为研究对象,所有研究对象均行HR-HPV分型检测、TCT及阴道镜下宫颈活检组织病理检查;以组织病理学检查结果为“金标准”(正常或炎症为阴性,其余诊断结果为阳性),分析HR-HPV分型检测、TCT及二者联合检测的阳性预测值、阴性预测值、灵敏度、特异度及准确性,并绘制2种检测与联合检测筛查宫颈癌前病变和宫颈癌的受试者操作特征(receiver operating characteristic,ROC)曲线。结果:150例患者中,组织病理学诊断结果为阴性者82例,阳性者68例。以病理学检查结果为“金标准”,HR-HPV分型检测的灵敏度为89.71%,特异度为74.39%,阳性预测值为74.39%,阴性预测值为89.71%,准确性为81.33%;TCT的灵敏度为63.24%,特异度为87.80%,阳性预测值为81.13%,阴性预测值为74.23%,准确性为76.67%。HR-HPV分型检测的灵敏度及阴性预测值高于TCT(P<0.05),TCT的特异度高于HR-HPV分型检测(P<0.05)。ROC曲线显示:HR-HPV分型检测的曲线下面积(area under the curve,AUC)为0.820(95%CI 0.750~0.878),TCT的AUC为0.755(95%CI 0.678~0.822),联合检测的AUC为0.813(95%CI 0.742~0.872)。结论:相比TCT,HR-HPV分型检测在宫颈癌前病变和宫颈癌筛查中有较好的诊断效能,HR-HPV联合TCT不能进一步提高筛查效能。

中药治疗产后血虚发热的临床及机理研究

目的探讨当归补血合四物汤加减治疗产后血虚发热的临床疗效，并探讨其作用机理。方法选取2013年3月至2016年9月我院门诊及病房诊断为产后血虚发热患者60例，随机分为2组。中药组30例，给予当归补血合四物汤加减口服治疗5天。对照组30例，给予抗生素静滴和琥珀酸亚铁口服治疗5天。分别观察2组患者退热时间、治疗前后体温、中医证候积分、白细胞介素（IL）-2、IL-6、血清促红细胞生成素（EPO）、血红蛋白及白细胞计数等指标。结果中药组的疗效优于对照组，差异有统计学意义（P〈0．05）。治疗后中药组及对照组体温、中医证侯积分、白细胞计数较治疗前均下降，血红蛋白均较治疗前升高，差异有统计学意义（P〈0．05或P〈0．01）；退热时间中药组快于对照组，差异有统计学意义（P〈0．01）。结论当归补血合四物汤加减治疗产后血虚发热疗效显著，且可降低IL-6水平和白细胞计数，升高血红蛋白水平，无副作用，值得临床推广应用。

二胎产妇配偶育儿胜任感与知觉压力现状及其影响因素分析

目的探讨二胎产妇配偶育儿胜任感与知觉压力状况的现状,并分析其影响因素,为进行有效的干预措施提供理论依据。方法采用横断面研究方法,选取2019年3—12月249名产后1周以内的二胎产妇的配偶为研究对象,采用一般资料调查表、中文版育儿胜任感量表(Chinese version of parenting sense of competence scale,C-PSOC)和中文版知觉压力量表(Chinese perceived stress scale,CPSS)对其进行调查,分析配偶的育儿胜任感及其知觉压力现状及其影响因素。结果二胎产妇配偶的育儿胜任感得分(63.39±4.84)分,处于中等偏高水平,知觉压力得分(30.73±4.31)分,处于较大压力。多元线性回归分析结果显示:一二胎性别差异是二胎产妇配偶育儿胜任感及知觉压力的影响因素(均P<0.05),文化程度为育儿胜任感的影响因素,年龄为知觉压力的影响因素(均P<0.05)。结论二胎产妇配偶的在育儿过程中具备了基本的效能与能力,但其知觉压力较大。根据其影响因素,护理人员应关注文化程度较低的人群,给予相关育儿知识技能的指导,并给予正确的性别观念引导,以便提高其育儿胜任感与降低其知觉压力。

Endothelium-specific SIRT1 overexpression inhibits hyperglycemia-induced upregulation of vascular cell senescence

The rapidly increasing prevalence of diabetes mellitus worldwide is one of the most serious and challenging health problems in the 21st century.Mammalian sirtuin 1(SIRT1) has been shown to decrease high-glucose-induced endothelial cell senescence in vitro and prevent hyperglycemia-induced vascular dysfunction.However,a role for SIRT1 in prevention of hyperglycemia-induced vascular cell senescence in vivo remains unclear.We used endothelium-specific SIRT1 transgenic(SIRT1-Tg) mice and wild-type(WT) mice to construct a 40-week streptozotocin(STZ)-induced diabetic mouse model.In this mode,42.9% of wild-type(WT) mice and 38.5% of SIRT1-Tg mice were successfully established as diabetic.Forty weeks of hyperglycemia induced significant vascular cell senescence in aortas of mice,as indicated by upregulation of expression of senescence-associated markers including p53,p21 and plasminogen activator inhibitor-1(PAI-1).However,SIRT1-Tg diabetic mice displayed dramatically decreased expression of p53,p21 and PAI-1 compared with diabetic WT mice.Moreover,manganese superoxide dismutase expression(MnSOD) was significantly downregulated in the aortas of diabetic WT mice,but was preserved in diabetic SIRT1-Tg mice.Furthermore,expression of the oxidative stress adaptor p66Shc was significantly decreased in aortas of SIRT1-Tg diabetic mice compared with WT diabetic mice.Overall,these findings suggest that SIRT1-mediated inhibition of hyperglycemia-induced vascular cell senescence is mediated at least partly through the reduction of oxidative stress.

Overexpression of a dominant-negative mutant of SIRT1 in mouse heart causes cardiomyocyte apoptosis and early-onset heart failure

SIRT1,a mammalian ortholog of yeast silent information regulator 2(Sir2),is an NAD+-dependent protein deacetylase that plays a critical role in the regulation of vascular function.The current study aims to investigate the functional significance of deacetylase activity of SIRT1 in heart.Here we show that the early postnatal hearts expressed the highest level of SIRT1deacetylase activity compared to adult and aged hearts.We generated transgenic mice with cardiac-specific expression of a dominant-negative form of the human SIRT1(SIRT1H363Y),which represses endogenous SIRT1 activity.The transgenic mice displayed dilated atrial and ventricular chambers,and died early in the postnatal period.Pathological,echocardiographic and molecular phenotype confirmed the presence of dilated cardiomyopathy.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling analysis revealed a greater abundance of apoptotic nuclei in the hearts of transgenic mice.Furthermore,we show that cardiomyocyte apoptosis caused by suppression of SIRT1 activity is,at least in part,due to increased p53acetylation and upregulated Bax expression.These results indicate that dominant negative form of SIRT1(SIRT1H363Y)overexpression in mouse hearts causes cardiomyocyte apoptosis and early-onset heart failure,suggesting a critical role of SIRT1 in preserving normal cardiac development during the early postnatal period.

SIRT1 suppresses PMA and ionomycin-induced ICAM-1 expression in endothelial cells

Intercellular adhesion molecule-1 (ICAM-1) plays an important role in the recruitment of leukocytes to the endothelium, which causes inflammation and initiation of atherosclerosis. We have previously shown that endothelium-specific over-expression of class III deacetylase SIRT1 decreases atherosclerosis. We therefore addressed the hypothesis that SIRT1 suppresses ICAM-1 expression in the endothelial cells. Here, we found that expression of SIRT1 and ICAM-1 was significantly induced by PMA and ionomycin (PMA/Io) in human umbilical vein endothelial cells (HUVECs). Adenovirus-mediated over-expression of SIRT1 significantly inhibited PMA/Io-induced ICAM-1 expression in HUVECs. Knockdown of SIRT1 by RNA interference (RNAi) resulted in increased expression of ICAM-1 in HUVECs. Luciferase report assay showed that over-expression of SIRT1 suppressed ICAM-1 promoter activity both in basic and in PMA/Io-induced conditions. We further found that SIRT1 was involved in transcription complex binding on the ICAM-1 promoter by chromatin immunoprecipitation (ChIP) assays. Furthermore, SIRT1 RNAi increased NF-κB p65 binding ability to the ICAM-1 promoter by ChIP assays. Overall, these data suggests that SIRT1 inhibits ICAM-1 expression in endothelial cells, which may contribute to its anti-atherosclerosis effect.

Activation of STAT3 stimulates AHSP expression in K562 cells

Studies on the chaperone proteinα-hemoglobin stabilizing protein(AHSP)reveal that abundant AHSP in erythroid cells enhance the cells’tolerance to oxidative stress imposed by excessα-hemoglobin in pathological conditions.However,the potential intracellular modulation of AHSP expression itself in response to oxidative stress is still unknown.The present study examined the effect and molecular mechanism of STAT3,an oxidative regulator,on the expression of AHSP.AHSP expression increased in K562 cells upon cytokine IL-6-induced STAT3 activation and decreased in STAT3 knock-down K562 cells.Regulation of AHSP in oxidative circumstance was then examined inα-globin-overloaded K562 cells,and real-time PCR showed strengthened expression of both AHSP and STAT3.ChIP analysis showed binding of STAT3 to AHSP promoter and binding was significantly augmented with IL6 stimulation and uponα-globin overexpression.Dual luciferase reporter assays of the wildtype and mutated SB3 element,an IL-6RE site,in the AHSP promoter in K562 cells highlighted the direct regulatory effect of STAT3 on AHSP gene.Finally,direct binding of STAT3 to SB3 site of AHSP promoter was confirmed with EMSA assays.Our work reveals an adaptive AHSP regulation mediated by the redox-sensitive STAT3 signaling pathway,and provides clues to the therapeutic strategy for AHSP enhancement.

Exploring CTCF and cohesin related chromatin architecture at HOXA gene cluster in primary human fibroblasts

Spatial expression patterns of homeobox(HOX) genes delineate positional identity of primary fibroblasts from different topographic sites. The molecular mechanism underlying the establishing or maintaining of HOX gene expression pattern remains an attractive developmental issue to be addressed. Our previous work suggested a critical role of CTCF/cohesin-mediated higher-order chromatin structure in RA-induced HOXA activation in human teratocarcinoma NT2/D1 cells. This study investigated the recruitment of CTCF and cohesin, and the higher-order chromatin structure of the HOXA locus in fetal lung and adult foreskin fibroblasts, which display complementary HOXA gene expression patterns. Chromatin contacts between the CTCF-binding sites were observed with lower frequency in human foreskin fibroblasts. This observation is consistent with the lower level of cohesin recruitment and 5′ HOXA gene expression in the same cells. We also showed that CTCF-binding site A56(CBSA56) related chromatin structures exhibit the most notable changes in between the two types of cell, and hence may stand for one of the key CTCF-binding sites for cell-type specific chromatin structure organization. Together, these results imply that CTCF/cohesin coordinates HOXA cluster higher-order chromatin structure and expression during development, and provide insight into the relationship between cell-type specific chromatin organization and the spatial collinearity.

Mutual inhibition between miR-34a and SIRT1 contributes to regulation of DNA double-strand break repair

DNA double-strand breaks are repaired through either non-homologous end joining(NHEJ) or homologous recombination repair(HRR) pathway.The well-characterized regulatory mechanisms of double-strand break repair(DSBR) are mainly found at the level of complicated repair protein interactions and modifications.Regulation of DSBR at the transcriptional level was also reported.In this study,we found that DSBR can be regulated by miR-34a at the post-transcriptional level.Specifically,miR-34a,which can be activated by DNA damages,represses DSBR activities by impairing both NHEJ and HRR pathways in cultured cells.The repression is mainly through targeting the critical DSBR promoting factor SIRT1,as ectopically expressed SIRT1 without 3'-UTR can rescue the inhibitory roles of miR-34a on DSBR.Further studies demonstrate that SIRT1 conversely represses miR-34a expression.Taken together,our data show that miR-34a is a new repressor of DSBR and the mutual inhibition between miR-34a and SIRT1 may contribute to regulation of DNA damage repair.

Mechanistic perspectives of calorie restriction on vascular homeostasis

Calorie restriction(CR)is a dietary regime based on low calorie intake.CR without malnutrition extends lifespan in a wide range of organisms from yeast to rodents,and CR can prevent and delay the onset of age-related functional decline and diseases in human and non-human primates.CR is a safe and effective intervention to reduce vascular risk factors in humans.In recent years,studies in rodents have provided mechanistic insights into the beneficial effects of CR on vascular homeostasis,including reduced oxidative stress,enhanced nitric oxide(NO)bioactivity,and decreased inflammation.A number of important molecules,including sirtuins,AMP-activated protein kinase,mammalian targets of rapamycin,endothelial nitric oxidase and their regulatory pathways are involved in the maintenance of vascular homeostasis.Evidence has shown that these pathways are responsible for many aspects of CR’s effects,and that they may also mediate the effects of CR on vasculature.