瞬时感受器电位离子通道香草素受体4在睾丸缺血再灌注损伤中对GC-1细胞增殖及凋亡的作用及其机制

目的探讨瞬时感受器电位离子通道香草素受体4(TRPV4)在睾丸缺血再灌注损伤(IRI)中对GC-1细胞增殖和凋亡的作用及其机制。方法建立睾丸GC-1细胞缺氧复氧模型,采用蛋白质印迹法(Western blot)检测不同复氧损伤时间点TRPV4的表达变化;分别采用噻唑蓝(MTT)实验、流式细胞术检测转染TRPV4对GC-1细胞增殖和凋亡的影响;采用Western blot法检测睾丸组织中转染TRPV4对GC-1细胞中半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3和细胞色素C(Cyt-C)表达的影响,组间比较采用t检验,多组间比较采用单因素方差分析。结果对照组及缺氧复氧组(0、6、12、24、48和72 h)TRPV4的蛋白表达水平分别为0.19±0.02、0.35±0.03、0.42±0.04、0.46±0.04、0.62±0.05、0.54±0.05、0.45±0.04。缺氧复氧组TRPV4表达显著高于未缺氧GC-1细胞的对照组,并在复氧24 h达到峰值(F=6.898,P<0.05),差异有统计学意义;缺氧复氧组中细胞增殖水平明显低于对照组(52.32±4.58比100.00±7.63,t=-9.280,P<0.05),差异有统计学意义,细胞凋亡水平高于对照组(15.60±1.72比4.08±0.87,t=10.352,P<0.05),差异有统计学意义;过表达TRPV4组中细胞增殖水平低于其对照组(23.65±3.98比51.35±4.67,t=-7.820,P<0.05),差异有统计学意义,细胞凋亡水平高于其对照组(26.93±2.15比14.62±1.68),t=7.814,P<0.05),差异有统计学意义;沉默TRPV4组中细胞增殖水平高于其对照组(72.49±6.21比53.18±5.14,t=4.150,P<0.05),差异有统计学意义,细胞凋亡水平低于其对照组(9.71±1.25比15.07±1.64,t=-4.502,P<0.05),差异有统计学意义。缺氧复氧组中Caspase-3和Cyt-C表达水平高于对照组(0.70±0.06比0.20±0.02,t=13.693,P<0.05;0.74±0.07比0.26±0.03,t=10.917,P<0.05),差异有统计学意义;过表达TRPV4组中Caspase-3和Cyt-C表达水平高于其对照组(1.25±0.11比0.69±0.07,t=7.439,P<0.05;1.38±0.14比0.72±0.07,t=7.303,P<0.05),差异有统计学意义;沉默TRPV4组中Caspase-3和Cyt-C表达水平低于其对照组(0.46±0.05比0.68±0.06,t=-4.879,P<0.05;0.45±0.05比0.72±0.06,t=-5.988,P<0.05),差异有统计学意义。结论TRPV4在GC-1细胞中高表达,其可能通过改变GC-1细胞的增殖和凋亡能力,从而影响睾丸IRI的发生发展。     

Mesenteric neovascularization during spring-mediated intestinal lengthening

BackgroundShort gut syndrome, a condition characterized by inadequate absorption of nutrients owing to decreased bowel length, has minimal avenues for treatment. We have proposed spring-mediated distraction enterogenesis to lengthen bowel in porcine jejunum as a treatment for short gut. We aim to evaluate the extent of mesenteric neovascularization in segments of lengthened bowel via spring-mediated enterogenesis.MethodsFemale juvenile Yucatan pigs underwent laparotomy and insertion of gelatin-encapsulated compressed nitinol springs, held in place with plication sutures, into the jejunum. At surgery and sacrifice, macroscopic mesenteric blood vessels were counted between the plication sites. Histologic samples of the mesentery were obtained to evaluate microscopic vasculature.ResultsA statistically significant increase in macroscopic mesenteric blood vessels was seen after intestinal lengthening (before: 1.9 ± 0.7 vessels, after: 4.7 ± 1.2 vessels, p = 0.001). A statistical significance is also seen in the density of arterioles (control: 3.0 ± 3.0 vessels/mm, spring: 7.0 ± 9.0 vessels/mm, p = 0.01) and venules (control: 4.0 ± 3.0 vessels/mm, spring: 8.0 ± 8.0 vessels/mm, p = 0.003).ConclusionIntestinal segments lengthened by intraluminal springs demonstrated total greater number of macroscopic vessels and microscopic blood vessels per length of mesentery as compared to control. This suggests local changes within the mesentery to recruit blood supply to growing intestine.Level of evidenceN/AType of studyTreatment study.     

Mechanical lengthening of porcine small intestine with decreased forces

Introductionshort bowel syndrome is marked by inadequate intestinal surface area to absorb nutrients. Current treatments are focused on medical management and surgical reconfiguration of the dilated intestine. We propose the use of spring-mediated distraction enterogenesis as a novel intervention to increase intestinal length. Given our previous success lengthening intestinal segments using springs with spring constant ~7 N/m that exerts 0.46 N or higher, we sought to determine the minimal force needed to lengthen porcine small intestinal segments, and to explore effects on intestine over time.MethodsJuvenile Yucatan pigs underwent laparotomy with enterotomy to introduce nitinol springs intraluminally (n = 21 springs). Bowel segments (control, spring-distracted) were retrieved on post-operative day (POD) 7 and 14, and lengths measured. Thickness of cross-sectional intestinal layers were measured using H&E, and submucosal collagen fiber orientation measured using trichrome stained sections.Resultsall pigs survived to POD7 and 14. Spring constants of at least 2 N/m exerting a minimum force of 0.10 N significantly lengthened intestinal segments (p <0.0001). The stronger the spring force, the greater the induced thickness of various intestinal layers at POD7 and 14. Collagen fiber orientation was also more disordered because of stronger springs.Conclusiona spring constant of approximately 2 N/m exerting 0.10 N and greater significantly lengthens intestinal segments and stimulates intestinal structural changes at POD7 and 14. This suggests a decreased force is capable of inducing spring-mediated distraction enterogenesis.     

Targeting NF-κB c-Rel in regulatory T cells to treat corneal transplantation rejection

The relevance of Tregs in the induction of tolerance against corneal allografts has been well established. Although it is well known that the conversion of Tregs into effector-like cells contributes to the loss of corneal immune privilege, the underlying mechanism is still not fully understood. Using heterologous penetrating keratoplasty model, we found that Tregs from corneal allograft rejected mice (inflam-Tregs) exhibit impaired function and characteristics of effector T cells. Further study showed that the expression of NF-κB c-Rel, a key mediator of effector T cell function, was significantly increased in inflam-Tregs. Mechanistic study revealed that elevated NF-κB c-Rel level in inflam-Tregs impaired Treg function through the promotion of inflammatory cytokine production and glycolysis. More importantly, we demonstrated that targeting NF-κB c-Rel was able to improve the immune suppressive function of inflam-Tregs in vitro and enhance the potential of them to suppress corneal transplantation rejection. Therefore, our current study identified NF-κB c-Rel as a key mediator of the conversion of Tregs into effector-like cells when under inflammatory environment.     

Predictive model containing PI-RADS v2 score for postoperative seminal vesicle invasion among prostate cancer patients

BackgroundSeminal vesicle invasion (SVI) is considered to be one of most adverse prognostic findings in prostate cancer, affecting the biochemical progression-free survival and disease-specific survival. Multiparametric magnetic resonance imaging (mpMRI) has shown excellent specificity in diagnosis of SVI, but with poor sensitivity. The aim of this study is to create a model that includes the Prostate Imaging Reporting and Data System version 2 (PI-RADS v2) score to predict postoperative SVI in patients without SVI on preoperative mpMRI.MethodsA total of 262 prostate cancer patients without SVI on preoperative mpMRI who underwent radical prostatectomy (RP) at our institution from January 2012 to July 2019 were enrolled retrospectively. The prostate-specific antigen levels in all patients were <10 ng/mL. Univariate and multivariate logistic regression analyses were used to assess factors associated with SVI, including the PI-RADS v2 score. A regression coefficient-based model was built for predicting SVI. The receiver operating characteristic curve was used to assess the performance of the model.ResultsSVI was reported on the RP specimens in 30 patients (11.5%). The univariate and multivariate analyses revealed that biopsy Gleason grade group (GGG) and the PI-RADS v2 score were significant independent predictors of SVI (all P<0.05). The area under the curve of the model was 0.746 (P<0.001). The PI-RADS v2 score <4 and Gleason grade <8 yielded only a 1.8% incidence of SVI with a high negative predictive value of 98.2% (95% CI, 93.0–99.6%).ConclusionsThe PI-RADS v2 score <4 in prostate cancer patients with prostate-specific antigen level <10 ng/mL is associated with a very low risk of SVI. A model based on biopsy Gleason grade and PI-RADS v2 score may help to predict SVI and serve as a tool for the urologists to make surgical plans.     

Value of preoperative urine white blood cell and nitrite in predicting postoperative infection following percutaneous nephrolithotomy: a meta-analysis

BackgroundTo evaluate to what degree preoperative urine white blood cell (WBC) and urine nitrite (NIT) values are predictive of postoperative infections following percutaneous nephrolithotomy (PCNL).MethodsA systematic literature search was performed of the PubMed, Embase, Cochrane Library, Wanfang Data, National Knowledge Infrastructure (CNKI), and China Science and Technology Journal Database (CSTJ or VIP) online databases to identify relevant studies that examined the predictive value of urine WBC or NIT as risk factors for post-PCNL infection, and the search was finished on February 28, 2020. Two independent reviewers screened the relevant studies, extracted necessary data from the eligible case-control studies (CCS), and assessed the quality of included studies through the Newcastle-Ottawa scale (NOS). RevMan 5.3 software and the Stata 16.0 software were used to complete the statistical analysis of data. Results are expressed as odds ratio (OR) with 95% confidence intervals (CIs).ResultsAccording to the statistical analysis of 12 eligible studies involving 6113 patients, positive urine WBC (WBC+: OR =3.86, 95% CI: 3.03–4.91, P<0.001) and positive NIT (NIT⁺: OR =7.81, 95% CI: 5.44–11.21, P<0.001) in preoperative tests were identified as independent risk factors for postoperative infections following PCNL.ConclusionsIn summary, as risk factors for postoperative infections, the presence of preoperative urine WBC+ and NIT+ should be evaluated as part of clinical procedure, in order to reduce infections of PCNL.     

凉山彝族自治州布拖县2010-2019年抗病毒治疗HIV/AIDS死亡的影响因素分析

目的 分析2010-2019年凉山彝族自治州布拖县抗病毒治疗HIV/AIDS的死亡影响因素,为今后制定可持续的抗病毒治疗策略提供参考依据。方法 采用病例对照研究方法,收集2010-2019年布拖县接受抗病毒治疗HIV/AIDS与死亡者基本和随访信息,按病例数2倍抽样组成对照组,采用logistic回归模型分析其死亡的影响因素。结果 研究对象为抗病毒治疗的HIV/AIDS 3 355例,死亡组1 179例,对照组共2 176例。其中,30~49岁占81.34%,男性占69.09%,彝族占99.55%,已婚或同居占91.12%,初中及以下文化程度占95.77%,农民占88.41%。多因素logistic回归分析结果显示,研究对象的死亡风险因素中,年龄≥50岁是18~29岁的5.08倍（95%CI：3.05~8.48）、女性是男性的0.70倍（95%CI：0.52~0.94）、注射吸毒传播途径是异性性传播途径的1.43倍（95%CI：1.06~1.91）、治疗前CD4⁺T淋巴细胞计数（CD4）≥350个/μl是CD4<200个/μl的0.38倍（95%CI：0.30~0.48）、最近1次使用含洛匹那韦/利托那韦（LPV/r）抗病毒治疗方案是司他夫定（d4T）+拉米夫定（3TC）+奈韦拉平（NVP）/依非韦伦（EFV）方案的0.04倍（95%CI：0.01~0.18）、耐药是不耐药的3.40倍（95%CI：2.13~5.42）,无病毒载量结果且未做耐药检测是不耐药的12.98倍（95%CI：10.28~16.40）。结论 年龄、性别、传播途径、治疗前CD4、最近1次抗病毒治疗方案、抗病毒治疗后耐药检测情况是布拖县接受抗病毒治疗HIV/AIDS的死亡影响因素。应扩大病毒载量和耐药检测覆盖面,科学更换抗病毒治疗方案,开展依从性教育和医务人员培训,降低抗病毒治疗HIV/AIDS死亡率。     

积分加权法评价磷化学烧伤实验病理的意义与应用

目的：积分加权法在病理组织学检查中的应用，方法：在常规病理描述基础上，采用积分加权法将病理结果予以定量，综合评价致伤后实验动物的主要脏器，结果：实验动物同等面积的磷酸烧伤各脏器积分降低，钙治疗后则显著改善，均与其它方面报道的资料一致，结论：积分加以数值使各脏器损伤程度一目了然，对不同脏器间的损伤情况有可比性，从而有助于了解主要受损脏器与总体的关系以及药物的治疗效果。