慢病毒介导的shRNA干扰大鼠施万细胞RSC96 MYH14基因的表达

目的 应用RNA干扰技术构建大鼠肌球蛋白重链14（MYH14）基因重组慢病毒载体并鉴定。方法 根据MYH14 mRNA序列设计合成单链引物形成双链寡核苷酸序列,连接入经AgeⅠ和BamHⅠ双酶切线性化的GV298慢病毒质粒载体中,菌液经PCR鉴定并测序验证。取测序正确的菌液提取质粒转染大鼠施万细胞RSC96,利用免疫荧光法观察转染效率,蛋白质印迹法筛选有效敲低MYH14的shRNA质粒,CCK-8法检测转染后RSC96细胞的活力。结果 合成3对MYH14-shRNA序列并将其克隆到GV298载体中,构建了重组质粒MYH14-shRNA1、2、3,经菌液PCR鉴定和测序筛选出测序正确的载体MYH14-shRNA1和MYH14-shRNA2。免疫荧光检测结果显示转染72 h时RSC96细胞荧光表达最强;蛋白质印迹法检测结果显示,与阴性对照（scramble序列）组相比,MYH14-shRNA2转染后RSC96细胞中MYH14蛋白的表达水平降低（0.57±0.15 vs 1.11±0.06,P<0.01）,而MYH14-shRNA1转染后MYH14蛋白表达水平差异无统计学意义（P>0.05）。MYH14-shRNA2转染RSC96细胞24 h后的细胞活力与阴性对照组相比差异无统计学意义（1.09±0.16 vs 1.00±0.15,P>0.05）。结论 成功构建大鼠MYH14基因重组慢病毒干扰载体,该载体能有效下调RSC96细胞中MYH14的表达。     

NDRG2 acts as a negative regulator downstream of androgen receptor and inhibits the growth of androgen-dependent and castration-resistant prostate cancer

Castration resistance is a major issue during castration therapy for prostate cancer and thus more effective treatment are needed for castration-resistant prostate cancer (CRPC). NDRG2 (N-Myc downstream regulated gene 2), a recently identified tumor suppressor, was previously shown to inhibit the proliferation and invasion of prostate cancer, but whether NDRG2 is involved in CRPC remains to be known. Because androgen receptor (AR) axis plays an important role in castration resistance, we evaluate the role of NDRG2 in AR signaling and CRPC. Immunohistochemistry examination of prostate cancer tissues demonstrated that the expression of NDRG2 is negatively correlated with that of AR and c-Myc. Furthermore, AR negatively regulates NDRG2, as well as alters levels of c-Myc and prostate specific antigen (PSA). Forced expression of NDRG2 significantly inhibits the in vitro growth of androgen-dependent and castration-resistant prostate cancer cells; this was accompanied by alterations in PSA, but not by those of AR and c-Myc. Finally, by mimicking castration therapy in a xenograft mouse model, we showed that lentivirus-mediated NDRG2 overexpression efficiently overcomes castration resistance. Thus, by acting as a negative regulator downstream of AR, NDRG2 may emerge as a potential therapy molecule for CRPC.     

Isolated Dislocation of the Posterior Tibial Tendon in an Amateur Snowboarder: A Case Report

Isolated dislocation of the posterior tibial tendon is an uncommon pathologic entity that typically occurs in the setting of acute trauma. The diagnosis remains challenging and is often delayed second to the rarity of the injury and symptoms similar to that of medial ankle sprains and other routinely diagnosed injuries about the ankle. The factors that predispose this tendon to dislocation include a hypoplastic retromalleolar groove, flexor retinaculum insufficiency, chronic repetitive trauma, and a structural abnormality from a previous medial malleolar fracture, or a combination thereof. Dislocation has also been cited as a complication of multiple local steroid injections and tarsal tunnel release. The mechanism of injury appears to involve forced dorsiflexion and eversion of the ankle when the posterior tibial tendon is contracted. Most cases do not respond well to conservative treatment and will require surgery to restore function and eliminate symptoms. We report a case of posterior tibial tendon dislocation related to a snowboarding injury and offer our technique for surgical correction.     

The contribution of physical inactivity and socioeconomic factors to type 2 diabetes in Nepal: A structural equation modelling analysis

Background and aimType 2 diabetes mellitus (T2DM) is emerging as a significant public health challenge in Nepal. Behavioural, social and economic changes are likely to play a part in the rise of this chronic disease, as they are in many developing countries. A better understanding of the relationship between physical activity (PA), socioeconomic factors and T2DM can inform the design of prevention programs. This study aimed to identify the path relationships between PA, socioeconomic position, anthropometric and metabolic variables and T2DM.Methods and resultsThis study analysed data from 1977 Nepalese adults aged 40–69 years from the cross-sectional WHO STEPS survey undertaken in 2013. The latent variable “PA” was created using the information on domains of PA while the latent variable “socioeconomic position” was created using the variables education, occupation and ethnicity. Participants’ fasting blood glucose was used to determine their diabetes status. Structural equation modelling was conducted, and correlations and adjusted regression coefficients are reported.Individuals with higher education, in paid employment and from advantaged ethnic groups were more likely to have T2DM. Waist circumference, triglycerides and hypertension were found to have a statistically significant positive direct effect on T2DM. PA had indirect effects on T2DM, mediated by waist circumference. The indirect effects of socioeconomic position on T2DM were mediated by body mass index, waist circumference, triglycerides and total cholesterol.ConclusionAmong Nepalese adults, higher socioeconomic position had a significant direct effect on T2DM, while both PA and higher socioeconomic position had significant indirect effects. Policies and programs to address T2DM in Nepal should address the factors contributing to unhealthy weight status, particularly among those of higher socioeconomic status.     

Triglyceride to high-density lipoprotein cholesterol ratio and risk of atherosclerotic cardiovascular disease in a Chinese population

Background and aimsTriglyceride (TG) to high-density lipoprotein cholesterol (HDL-C) ratio may play a role in predicting cardiovascular events. We aimed to prospectively explore the association between the TG/HDL-C ratio and atherosclerotic cardiovascular disease (ASCVD), ischemic stroke, as well as coronary heart disease (CHD) in a Chinese population.Methods and resultsThis prospective cohort study included 9368 participants from four Chinese populations in the People's Republic of China–United States of America (PRC-USA) Collaborative Study of Cardiovascular and Cardiopulmonary Epidemiology. Over a follow-up period of 20 years, 624 cases of ASCVD events including 458 ischemic stroke events and 166 CHD events were recorded. The relationship between the TG/HDL-C ratio and the endpoints was evaluated through multivariate Cox proportional hazard models adjusted for potential confounding variables, including age, sex, urban or rural residence, northern or southern China, occupational type, education, physical exercise, smoking status, drinking status, body mass index, hypertension, high low-density lipoprotein cholesterol, diabetes, and antihypertensive medication use at baseline. With the lowest TG/HDL-C tertile as the reference, the middle and highest tertiles had the hazard ratios (HRs) and 95% confidence intervals (CIs) of 1.13 (0.91, 1.40), 1.36 (1.10, 1.67) respectively for ASCVD (p for trend = 0.0028), and 1.19 (0.93, 1.54),1.47 (1.15, 1.87) respectively for ischemic stroke (p for trend = 0.0016). However, no significant association was found for CHD events.ConclusionTG/HDL-C ratio was positively associated with the risk of ASCVD and ischemic stroke events in the Chinese population.     

Sustained clinical safety and efficacy of a biodegradable‐polymer coated sirolimus‐eluting stent in “real‐world” practice: Three‐year outcomes of the CREATE (multi‐center registry of EXCEL biodegradable polymer drug eluting stents) study

Background: The CREATE is a post‐marketing surveillance multicenter registry that demonstrated satisfactory angiographic and clinical (at 18 months) outcomes of a biodegradable polymer based sirolimus‐eluting stent (EXCEL, JW Medical System, Weihai, China) for the treatment of patients in routine clinical practice. Objectives: To evaluate the three‐year clinical safety and efficacy outcomes in patients enrolled in the CREATE study. Methods: A total of 2077 all comers have been enrolled in the CREATE study at 59 centers from four countries. Recommended antiplatelet regimen was clopidogrel and aspirin for six months followed by chronic aspirin therapy. The prespecified primary outcome was the rate of major adverse cardiac events (MACE) at 12, 18, and 36 months. Results: Clinical follow‐up was completed in 2025 (97.5%) patients at three years. The average duration of clopidogrel treatment was 199.8 ± 52.7 days and 80.5% of discharged patients discontinued clopidogrel at six months. The cumulative rate of MACE was 4.5% and the rate of stent thrombosis was 1.53% at three years. At six months to three years, prolonged clopidogrel therapy (>6 months) was not beneficial in reducing cumulative hazards of MACE (3.4% vs. 3.1%, log rank P = 0.725) or stent thrombosis (1.5% vs. 0.6%, log rank P = 0.053). Conclusions: This study demonstrates sustained three‐year clinical safety and efficacy of biodegradable polymer‐based sirolimus‐eluting stents when used with six months of dual antiplatelet therapy in a “real‐world” setting. © 2011 Wiley Periodicals, Inc.     

胃肠道小间质瘤55例临床分析

目的：探讨胃肠道小间质瘤临床特点和诊治方法。方法回顾性分析解放军总医院2004年10月-2010年4月接受手术治疗的55例胃肠道小间质瘤患者的临床资料,同时进行随访分析。结果55例中有临床症状者占58.2%;胃部间质瘤占80.0%,其中胃上部间质瘤占52.7%。 Fletcher危险度分级极低和低度占92.7%,肿瘤直径≤1 cm危险度分级更低,Ki-67阳性与危险度分级无关。肿瘤切除采取微创手术者33例（微创手术组）,其中腹腔镜下切除17例,内镜下切除14例,双镜联合局部切除2例;采取传统开放手术者22例（开放手术组）,其中局部切除18例,近端胃切除3例,胰十二指肠切除1例。微创手术组较开放手术组术中出血量及术后开始进食时间差异均有统计学意义（P〈0.01,P〈0.05）。38例获得随访,随访时间26~104个月,中位时间41个月,仅1例胃体间质瘤内镜下切除术后22个月复发。结论胃肠道小间质瘤患者仅半数有临床症状,肿瘤多生长于胃上部,危险度分级及复发风险低,Ki-67不能作为判断胃肠道小间质瘤预后的指标。微创手术治疗安全、有效。     

Mice with ablated adult brain neurogenesis are not impaired in antidepressant response to chronic fluoxetine

The neurogenesis hypothesis of major depression has two main facets. One states that the illness results from decreased neurogenesis while the other claims that the very functioning of antidepressants depends on increased neurogenesis. In order to verify the latter, we have used cyclin D2 knockout mice (cD2 KO mice), known to have virtually no adult brain neurogenesis, and we demonstrate that these mice successfully respond to chronic fluoxetine. After unpredictable chronic mild stress, mutant mice showed depression-like behavior in forced swim test, which was eliminated with chronic fluoxetine treatment, despite its lack of impact on adult hippocampal neurogenesis in cD2 KO mice. Our results suggest that new neurons are not indispensable for the action of antidepressants such as fluoxetine. Using forced swim test and tail suspension test, we also did not observe depression-like behavior in control cD2 KO mice, which argues against the link between decreased adult brain neurogenesis and major depression.     

军队文职人员护理骨干执行卫勤保障任务现状的调查

目的了解军队文职人员护理骨干在执行卫勤保障任务中存在的问题。方法采用自制的调查问卷对参加全军第3批文职人员护理骨干任职教育培训班的83名文职人员进行调查。结果文职人员护理骨干在执行卫勤保障任务中存在的主要问题排在前三的依次是:执行大型卫勤保障任务的机会太少、平时科室工作繁忙、工作与训练矛盾突出,对卫勤保障任务的目的、目标及自身的职责范围不是很明确。结论相关部门应适当增加文职人员执行大型卫勤保障任务的机会,部门之间搞好协调工作,缓解工作和训练的矛盾,在卫勤保障中明确任务的目的、目标和参训人员的职责范围,以提高文职人员的卫勤保障能力。     

CORRECTION

Background  Connective tissue growth factor (CTGF) is a secreted protein containing several domains that mediate interactions with growth factors, integrins and extracellular matrix components. CTGF plays an important role in extracellular matrix production by its ability to mediate collagen deposition during wound healing. CTGF also induces neovascularization in vitro, suggesting a role in angiogenesis in vivo. We herein evaluated whether CTGF was required for extracellular matrix synthesis of meniscal fibrochondrocytes and/or angiogenesis during the repair of meniscal tears.

Methods  Meniscal fibrochondrocytes were isolated from the inner-1/2 of rabbit meniscus by trypsin collagenase treatment and further treated with 100 ng/ml CTGF in vitro. Characterization of fibrochondrocytes was identified by flow cytometry analyzing CD31, CD44, CD45 and CD105, and was further tested by type II collagen immunocytochemistry. Changes in gene expression of meniscal fibrochondrocytes were monitored by quantitative real-time polymerase chain reaction. Histological sections prepared from a 3-mm portion of a longitudinal tearing defect in the middle of the rabbit meniscus were subjected to fluorescence-immunohistochemistry analysis at 1, 4 and 10 weeks following surgical treatment with 1.5 μg of CTGF/fibrin-glue composites.

Results  Quantitative RT-PCR assay showed that types I and II collagen and vascular endothelial growth factor mRNA expression in the 100 ng/ml CTGF group were remarkably enhanced as compared to levels in the no-dose group at 14 days ((2.38±0.63) fold, (2.96±0.87) fold, (2.14±0.56) fold, respectively). Likewise, fluorescence-immunohistochemical analysis revealed that in the group implanted with CTGF-fibrin glue, types I and II collagen, as well as the capillaries, completely filled the defect by 10 weeks, postoperatively. In contrast, only soft tissue repair occurred when PBS-fibrin glue was implanted.

Conclusions  These findings suggest that CTGF can significantly promote extracellular matrix deposition (types I and II collagen) within the meniscal avascular zone; CTGF can greatly heighten the expression of vascular endothelial growth factor activity simultaneously in vivo, further enhancing the repair of meniscal tears in the avascular zone.