乙型肝炎病毒全基因在肝癌细胞中的转染与表达

目的 建立乙型肝炎病毒（HBV）复制状态模型。方法 体外连接HBV DNA获得6.4 kb的双拷贝DNA片段,然后将其插入pcDNA3载体的Eco RV位点,通过聚合酶链反应（PCR）和酶切筛选获得头尾串联双拷贝HBV全基因重组真核表达质粒,再通过脂质体介导法转染肝癌细胞。结果 通过G418筛选,对阳性克隆细胞进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和Western免疫印迹分析,证明已将HBV全基因重组真核表达质粒转染至肝癌细胞并获得稳定表达。结论 通过脂质体介导法将HBV全基因转染至肝癌细胞,成功建立了HBV复制状态的细胞模型。     

Mechanism of miR-365 in regulating BDNF-TrkB signal axis of HFD/STZ induced diabetic nephropathy fibrosis and renal function

Purpose

Diabetic nephropathy (DN) is one of the most serious complications of diabetes that leads to decline of renal function. Although numerous studies have revealed that microRNAs (miRNAs) play essential roles in the progression of DN, whether miR-365 is involved remains elusive.

Methods

The successful construction of DN model was confirmed by ELSIA, hematoxylin–eosin (HE) and Masson staining assay. The expression of miR-365 was detected through RT-qPCR. The levels of BDNF, p-TrkB, α-smooth muscle actin (SMA), collagen IV (Col.IV), transforming growth factor-β1 (TGF-β1), tumor necrosis factor α (TNF-α), and interleukin-6 (IL-6) were evaluated by western blot, IF or ELISA assays. Luciferase reporter assay was used to detect the interaction between miR-365 and BDNF.

Results

The DN mice model was induced by streptozotocin (STZ). Then miR-365 expression was found to upregulate in tissues of DN rat. Furthermore, elevated expression of miR-365 was found in high glucose (HG)-treated HK-2 cells. Silencing of miR-365 suppressed the accumulation of ECM components and secretion of inflammatory cytokines in HK-2 cells. In addition, it was demonstrated that miR-365 could target BDNF. The protein levels of BDNF and p-TrkB were negatively regulated by miR-365 in HK-2 cells. Moreover, inhibition of miR-365 suppressed the levels of SMA, Col.IV, TGF-β1, TNF-α, and IL-6, indicating the renal fibrosis was inhibited by miR-365 knockdown.

Conclusion

MiR-365 could regulate BDNF-TrkB signal axis in STZ induced DN fibrosis and renal function. The results of the current study might provide a promising biomarker for the treatment of DN in the future.

     

Efficacy of testosterone replacement therapy for treating metabolic disturbances in late-onset hypogonadism: a systematic review and meta-analysis

International Urology and Nephrology - Late onset hypogonadism (LOH) is an age-dependent reduction of testosterone associated with alterations of metabolic profile, including glucose control,...     

Panax Notoginseng Saponins suppresses TRPM7 via the PI3K/AKT pathway to inhibit hypertrophic scar formation in vitro

BackgroundHypertrophic scar (HS) formation, a type of dermal fibroproliferative condition, is a frequent complication in wound healing resulting from burns, severe trauma, and surgical procedures. The effects of Panax Notoginseng Saponins (PNS) on the HS formation remain relatively under-explored. Hence, this study was intended to interrogate anti-apoptosis and anti-fibrosis effects of PNS on the hypertrophic scar fibroblasts (HSFs) during HS formation and assess the involvement of TRPM7 and PI3K/AKT signaling pathway.MethodsUsing MTT and CCK-8 assays, we evaluated cell cytotoxicity and cell viability. Collagen I/III (col 1/3) and α-SMA expression levels were assessed through immunofluorescence and western blot, and cell migration, cell apoptosis and cell cycle were examined with applications of wound healing, TUNEL staining and flow cytometry. TRPM7, PI3K/AKT, TGF-β1 and related-proteins were quantified using RT-qPCR and western blot.ResultsPNS administration could suppress TRPM7 expression and the viability of HSFs in a dose-dependent manner. Moreover, PNS could restrain the HS formation and ECM deposition by decreasing col 1/3 and α-SMA synthesis, suppressing cell migration, and boosting apoptosis and G1 arrest. Notably, this study revealed that PNS inhibited PI3K/AKT activation in HSFs. Besides, knockdown of TRPM7 enhanced therapeutic effects of PNS on HSFs, but overexpression markedly reversed above mentioned effects of PNS on HSFs.ConclusionThis study suggested that PNS hampered scar formation might via inhibiting ECM and stimulating cell apoptosis by modulating the PI3K/AKT signaling. Overall, these findings in the present study could support the use of PNS for preventing HS formation, and TRPM7 may be a novel molecular target for treating HS.     

多西他赛+卡铂联合曲妥珠单抗方案对早期人表皮生长因子受体2阳性乳腺癌的新辅助治疗效果

目的探讨多西他赛+卡铂联合曲妥珠单抗(TCH)方案对早期人表皮生长因子受体2(HER2)阳性乳腺癌的新辅助治疗效果。方法回顾性分析2013年1月至2018年12月北京大学第一医院乳腺疾病中心经治的522例早期HER2阳性乳腺癌患者的临床资料,占同期收治早期浸润性乳腺癌患者的21.80%(522/2 394)。其中113例接受TCH方案进行新辅助治疗,年龄[M(QR)]52(13)岁(范围:23~69岁)。记录TCH方案新辅助治疗后病理完全缓解(pCR,ypT0N0M0期)的例数,采用Miller-Payne标准进行病理学评价。采用Kaplan-Meier法计算无病生存率和总体生存率,采用Log-rank检验比较组间生存差异。结果接受曲妥珠单抗规范治疗患者(294例)的无病生存率优于未规范治疗患者(177例)(84.4%比72.4%,χ²=4.095,P=0.046)。发生3~4级不良反应的患者占全部患者的15.9%(18/113),包括3~4级中性粒细胞减少12例,腹泻6例。31例患者获得pCR(ypT0N0M0),pCR率为27.4%(31/113)。pCR患者与非pCR患者的无病生存率和总体生存率无差异(91.8%比85.0%,92.5%比90.5%,P值均>0.05)。病理学评价为G4~5的患者无病生存率优于G1~3患者(89.6%比81.5%,χ²=5.340,P=0.021),而总体生存率的差异无统计学意义(91.4%比89.1%,χ²=1.008,P=0.315)。结论早期HER2阳性乳腺癌采用TCH方案行新辅助治疗的效果较好,新辅助治疗后病理学评价为G4~5的患者的无病生存率更高。     

Bone Radiomics Score Derived From DXA Hip Images Enhances Hip Fracture Prediction in Older Women

Dual-energy X-ray absorptiometry (DXA)-based bone mineral density testing is standard to diagnose osteoporosis to detect individuals at high risk of fracture. A radiomics approach to extract quantifiable texture features from DXA hip images may improve hip fracture prediction without additional costs. Here, we investigated whether bone radiomics scores from DXA hip images could improve hip fracture prediction in a community-based cohort of older women. The derivation set (143 women who sustained hip fracture [mean age 73 years, time to fracture median 2.1 years] versus 290 age-matched women [mean age 73 years] who did not sustain hip fracture during follow-up [median 5.5 years]) were split into the train set (75%) and the test set (25% hold-out set). Among various models using 14 selected features out of 300 texture features mined from DXA hip images in the train set, random forest model was selected as the best model to build a bone radiomics score (range 0 to 100) based on the performance in the test set. In a community-based cohort (2029 women, mean age 71 years) as the clinical validation set, the bone radiomics score was calculated using a model fitted in the train set. A total of 34 participants (1.7%) sustained hip fracture during median follow-up of 5.4 years (mean bone radiomics score 40 ± 16 versus 28 ± 12 in non-fractured, p < 0.001). A one-point bone radiomics score increment was associated with a 4% elevated risk of incident hip fracture (adjusted hazard ratio [aHR] = 1.04, p = 0.001) after adjustment for age, body mass index (BMI), previous history of fracture, and femoral neck T-score, with improved model fit when added to covariates (likelihood ratio chi-square 10.74, p = 0.001). The association between bone radiomics score with incident hip fracture remained robust (aHR = 1.06, p < 0.001) after adjustment for FRAX hip fracture probability. Bone radiomics scores estimated from texture features of DXA hip images have the potential to improve hip fracture prediction. © 2021 American Society for Bone and Mineral Research (ASBMR).     

全麻术后苏醒期患者早期饮水的研究进展

综述全麻术后苏醒期患者早期饮水的必要性与可行性,重点分析早期饮水的实施要素,主要包括饮水的对象评估、种类及方式、饮水量及间隔时间和安全性,以期为我国全麻术后苏醒期患者开展早期饮水临床实践提供参考.     

组蛋白去甲基化酶含有Jumonji结构域的蛋白质2D在胃癌组织的表达及其临床意义

目的观察含有Jumonji结构域的蛋白质2D(JMJD2D)在胃癌组织的表达并探讨其临床意义。方法收集2013年1月至2014年12月期间来自中国科学院大学附属肿瘤医院(浙江省肿瘤医院)手术治疗并经病理诊断证实的133例胃腺癌标本。采用免疫组织化学法检测133例胃癌组织及其配对的癌旁正常组织中JMJD2D的表达,应用SPSS-22统计软件分析其临床意义及与预后的关系,采用比例风险回归模型(proportional hazards model,Cox模型)多因素回归分析JMJD2D蛋白表达及其他病理参数对胃癌患者预后的预测价值。结果胃癌组织中JMJD2D的高表达率为75.9%(101/133),显著高于癌旁正常胃组织(高表达率2.3%,3/133,χ²=64.821,P<0.01),差异有统计学意义。肿瘤组织中JMJD2D高表达与患者幽门螺杆菌感染状态(χ²=22.163,P<0.01)、肿瘤分化程度(χ²=7.022,P<0.05)、浸润深度(χ²=5.061,P<0.05)、淋巴结转移状态(χ²=14.123,P<0.01)、临床TNM分期(χ²=23.194,P<0.01)显著相关,而与患者性别、肿瘤诊断时年龄、肿瘤部位和大小无相关(χ²=0.072、1.451、2.562、1.383,P值均>0.05)。普兰-迈耶(Kaplan-Meier)生存分析显示,JMJD2D高表达的胃癌患者与正常/低表达患者中位生存时间分别为34个月和65个月,两者差异有统计学意义(P<0.05)。比例风险回归模型(proportional hazards model,Cox模型)多因素回归分析表明JMJD2D高表达是胃癌患者独立预后因素[危险比(HR)=2.38,P<0.01]。结论JMJD2D在胃癌组织中高表达,且与胃癌进展及患者预后显著相关。     

腹腔镜结直肠癌手术中转开腹列线图预测模型的构建

目的分析腹腔镜结直肠癌手术发生中转开腹的危险因素,建立列线图预测模型指导临床决策,并分析其短期预后影响。方法收集2017年5月至2020年5月西安交通大学第一附属医院行腹腔镜手术的764例结直肠癌患者的病例资料,分为中转开腹组(60例)及腹腔镜组(704例)。采用Logistic回归分析中转开腹的独立危险因素,并用R语言构建中转开腹列线图预测模型。通过受试者工作特征曲线(ROC)下面积评估列线图预测模型效能。通过绘制校准图进行一致性检验。并比较两组患者术中及术后恢复情况。结果本研究共纳入患者764例,其中60例发生中转开腹,中转开腹率为7.9%(60/764)。肿瘤位置位于直肠[比值比(OR)=1.846,P<0.05)]、美国麻醉医生协会(ASA)评分Ⅲ、Ⅳ级(OR=2.381,P<0.05)、腹部手术史(OR=3.652,P<0.01)、肿瘤长径≥5 cm(OR=2.704,P<0.05)和体重指数(OR=1.109,P<0.05)是发生中转开腹的独立危险因素,上述数据差异均有统计学意义。成功构建列线图预测模型,模型ROC曲线下面积为0.794。校正曲线显示预测结果与实际结果有较好的重合度。中转开腹组手术时间高于腹腔镜组(210 min比190 min,Z=-2.670,P<0.05);中转开腹组出血量高于腹腔镜组(170 ml比120 ml,Z=-6.018,P<0.01);中转开腹组术后住院天数高于腹腔镜组(10 d比9 d,Z=-2.134,P<0.05)。中转开腹组肠梗阻发生率高于腹腔镜组[10.0%(6/60)比4.3%(15/704),χ²=4.055,P<0.05];伤口感染率高于腹腔镜组[15.0%(9/60)比4.7%(33/704),χ²=11.318,P<0.05],上述数据差异均有统计学意义。结论肿瘤位置位于直肠、ASA评分Ⅲ、Ⅳ级、腹部手术史、肿瘤长径≥5 cm和体重指数是腹腔镜结直肠癌手术发生中转开腹的独立危险因素,据此建立的预测模型具有可靠的预测能力。     

Pediatric Femoral Shaft Fracture: An Age-Based Treatment Algorithm

PurposeFractures of the femoral shaft in children are common. The rates of bone growth and remodeling in children vary according to their ages, which affect their respective management. MethodsThis paper evaluates the incidence and patterns of pediatric femoral shaft fracture and the current concepts of treatments available.ResultsThe type of fracture—closed or open; stable or unstable—needs to be taken into account. Child abuse should be suspected in fractures sustained by infants. For younger children, non-surgical management is preferred, which include Pavlik harness (< 6 months old) and early spica casting (6 months to 6 years old). Older children (> 6 years old) usually benefit from surgical treatments as outcomes of non-surgical alternatives are worse and are associated with prolonged recovery times. These operative measures for older children that are 6–12 years old include elastic stable intramedullary nailing and submuscular plating. Factors to be considered when devising an appropriate intervention include body mass, location of injury, and nature of fracture. For adolescent and skeletally mature teenagers (> 12 years old), rigid antegrade entry intramedullary fixation is indicated. In the event of open fractures or polytrauma, external fixation should be considered as a temporary treatment method for initial fracture stabilization.ConclusionAn age-based and evidence-based algorithm has been proposed to guide surgeons in the process of evaluating an appropriate treatment.