应用 Ilizarov 技术胫骨横向骨搬移术治疗合并全身性炎症反应综合征的重度糖尿病足

目的探讨应用 Ilizarov 技术胫骨横向骨搬移术治疗合并全身性炎症反应综合征（systemic inflammatory response syndrome，SIRS）的重度糖尿病足（Wagner 3～5 级）的临床疗效。方法2014 年 8 月—2017 年 12 月，采用 Ilizarov 技术胫骨横向骨搬移术治疗合并 SIRS 的重度糖尿病足患者 33 例。其中男 27 例，女 6 例；年龄 34～79 岁，平均 60.6 岁。均患 2 型糖尿病，糖尿病病程 1～28 年，平均 10 年。糖尿病足病程 1～12 个月，平均 2.7 个月。根据 Wagner 分级，3 级 8 例，4 级 23 例，5 级 2 例。术后观察患足创面愈合情况, 并计算保肢率。评估体温、心率、呼吸频率、白细胞计数、红细胞沉降率和 C 反应蛋白浓度变化。测量足背皮肤温度；行疼痛视觉模拟评分（VAS）评价患足疼痛改善情况。结果术后 33 例患者均获随访，随访时间 3～30 个月，平均 14.1 个月。足部溃疡全部愈合，愈合时间 3～12 个月，平均 5.3 个月；保肢率 100%。术后体温、心率、呼吸频率、白细胞计数、红细胞沉降率和 C 反应蛋白浓度均较术前显著下降（P<0.05）。术后 1 个月患足皮温为（32.64±2.17）℃，较术前的（31.28±1.99）℃ 显著改善（t=0.05，P=0.00）；但与健侧皮温（32.46±2.10）℃ 比较差异无统计学意义（t=2.04，P=0.41）。术后 1 个月 VAS 评分为（2.4±0.7）分，较术前的（4.3±0.8）分显著改善（t=3.10，P=0.00）。 结论Ilizarov 技术胫骨横向骨搬移术能有效治疗合并 SIRS 的重度糖尿病足，可促进患足溃疡愈合，避免截肢。     

中文版颈部结局评分评估颈痛患者反应度的研究

目的探讨中文版颈部结局评分（NOOS-C）的反应度，为颈痛患者干预效果的评估提供可靠工具。方法对 NOOS 量表进行跨文化调适形成 NOOS-C。以 2016 年 9 月—2017 年 5 月收治并符合纳入标准的 80 例颈痛患者作为研究对象，干预前后使用 NOOS-C 和中文版颈椎功能障碍指数（NDI-C）进行评估，其中 71 例患者完成问卷调查。采用配对t 检验比较干预前后 NOOS-C 各维度评分及总分差异。计算 NOOS-C、NDI-C 评分干预前后差值（变化值），并基于变化值计算效应大小（effect size，ES）、标准化反应均数（standardized response mean，SRM），分析量表内部反应度。干预前后 NOOS-C、NDI-C 评分行 Spearman 相关分析，分析量表外部反应度。 结果干预前后 NOOS-C 评分中除活动评分比较差异无统计学意义（P>0.05）外，症状、睡眠障碍、日常活动和疼痛、日常生活参与度和总分比较，差异均有统计学意义（P<0.05）。NDI-C 指数变化值为–12.11%±17.45%、ES 为 0.77、SRM 为 0.69；NOOS-C 分别为（13.74±17.22）分、0.83、0.80。相关性分析显示，NOOS-C 与 NDI-C 干预前后均成负相关（r=–0.914，P=0.000； r=–0.872，P=0.000）。 结论NOOS-C 具有良好的反应度。     

Per and neuropeptide expression in the rat suprachiasmatic nuclei: compartmentalization and differential cellular induction by light

Per1 and Per2, two clock genes rhythmically expressed in the suprachiasmatic nucleus (SCN), are implicated in the molecular mechanism of the circadian pacemaker and play a major role in its entrainment by light. To date, it is not known if every cell of the SCN, a heterogeneous structure in respect of neuropeptide content, expresses clock genes equally. The aim of this study was to identify, by single and double non-radioactive and/or radioactive hybridizations, the cell types (AVP, VIP and GRP) expressing Per1 or Per2 in the SCN of rats, (1) when Per are highly expressed during the daytime, and (2) after induction of Per expression by a light pulse at night. Our results indicate that, during the daytime, Per1 and Per2 genes are both mainly expressed in the AVP cells of the dorso-median part of the SCN, whereas only a few VIP cells in the ventral part of the SCN exhibit Per gene expression. In contrast, following a light pulse at night, there is differential induction of the two Per genes. Per1 expression essentially occurs in the ventro-lateral GRP cells, while Per2 expression is not restricted to the retinorecipient part of the SCN as it also occurs in AVP cells. Altogether, our results suggest that Per1 and Per2 are mainly expressed in AVP cells during the daytime and suggest that GRP cells play an important role in resetting of the clock by light.     

心脏不停跳二尖瓣置换术对心肌内质网应激相关因子表达影响的研究及意义

【摘要】 目的 探讨心脏不停跳二尖瓣置换术（MVR）对心肌内质网应激（ERS）相关因子GRP78/Caspase-12表达的影响以及对心肌保护的意义。方法 选取择期行MVR的风湿性心脏病（RHD）二尖瓣狭窄（MS）患者40例,随机分为心脏不停跳组（BH组,n=20）与心脏停跳组（CA组,n=20）,分别于体外循环（CPB）转流开始前（T0）、主动脉阻断30 min后（BH组为CPB开始后30 min,T1）及缝合右心房前（T2）收集右心房心肌组织,通过rt-PCR法检测两组心肌GRP78和Caspase-12的表达,同时采用免疫组织化学法检测两组心肌GRP78和Caspase-12阳性着色情况。结果[结果部分应列举主要数据,并修改英文摘要] 两组GRP78mRNA表达：CA组T0（0.088±0.009）、T1（0.193±0.024）、T2（0.516±0.037）,BH组T0（0.085±0.008）、T1（0.189±0.022）、T2（0.229±0.038）,两组GRP78mRNA在T1时段表达量均增高（P＜0.05）,在T2时段表达量增高且CA组表达量高于BH组（P＜0.05）;两组Caspase-12mRNA表达：CA组T0（0.205±0.037）、T1（0.341±0.032）、T2（0.912±0.051）,BH组T0（0.198±0.035）、T1（0.209±0.035）、T2（0.307±0.063）,CA组Caspase-12 mRNA的表达量在T1、T2时段均增高（P＜0.05）;BH组仅在T2时段表达增高（P＜0.05）;CA组在T1、T2时间段Caspase-12 mRNA表达量均高于BH组（P＜0.05）。T2时段CA组与BH组GRP78蛋白免疫组化染色结果为（62.74±8.12）和（35.59±4.09）,CA组高于BH组（P＜0.05）;T2时段CA组与BH组Caspase-12蛋白免疫组化染色为（69.60±5.54）和（26.48±2.29）,CA组高于BH组（P＜0.05）。结论 心脏不停跳MVR能够通过减轻心肌ERS反应提高手术过程中的心肌保护。     

Isometric handgrip training improves local flow-mediated dilation in medicated hypertensives

Increased HSP expression in response to acute exercise is well documented in animal studies, and there is growing evidence that similar responses occur in man. In general, many human exercise studies have investigated the HSP response to low force continuous activity, while the knowledge about the HSP response to high force intermittent type of activity, like weight training, is so far sparse. In addition, most studies have used untrained subjects, and a common observation is that acute low force continuous activity in untrained individuals increases the HSP expression in these individuals. The main scope of this study was to investigate the HSP response in very well trained males subjected to longitudinal high intensity exercise, and if this response was dependent on exercise modality [i.e. eccentric (ECC) or concentric (CON) contractions]. Very well trained males performed progressive strength training consisting of either high force ECC or high force CON elbow flexions 2–3 times a week for 12 weeks. Compared with pre-exercise levels, HSP72 expression decreased by 46.1% (P<0.05) after CON contractions. GRP75 expression was unchanged after ECC or CON contractions, while ubiquitin expression decreased by 19.9% (P<0.02) after ECC contractions. These findings imply that chronic, intensive exercise may attenuate the HSP response in well-trained males.     

HSPA5 promoter polymorphisms and risk of Parkinson's disease in Taiwan

Endoplasmic reticulum (ER) stress induced by misfolded proteins has been implicated in Parkinson's disease (PD) pathogenesis. A malfunction of unfolded protein response (UPR) to ER stress can result in PD as well as other neurodegenerative diseases. Heat shock 70 kDa protein 5 (HSPA5) is one of the UPR chaperones reactive to ER stress to block the apoptotic process. HSPA5 promoter polymorphisms -415 G/A (rs391957), -370 C/T (rs17840761) and -180 del/G (rs3216733) and their derived haplotypes may affect promoter activity of the gene. This study examines whether these HSPA5 promoter polymorphisms are associated with the risk of Taiwanese PD and the age of disease onset using a case-control study. Polymorphisms -415 G/A and -180 del/G were completely linked in our population (D'=1.00, Delta(2)=1.00). The genotype or allele frequency distribution of each HSPA5 polymorphism was not significantly different between the controls (n=341) and the PD patients (n=393). Neither the linked -415 G/A and -180 del/G nor -370 C/T polymorphism influences PD onset age. Our data suggest that the HSPA5 -415 G/A, -370 C/T, and -180 del/G polymorphisms are unlikely to play a major role in risk of developing PD in Taiwan.     

促红细胞生成素对横纹肌溶解大鼠急性肾损伤时内质网应激相关蛋白的影响

目的探讨促红细胞生成素(EPO)能否通过减轻内质网应激相关蛋白葡萄糖调节蛋白78(GRP78)和C/EBP同源蛋白(CHOP)的表达以及减轻横纹肌溶解致急性肾损伤。方法将SD大鼠随机分为4组:对照组(control,n=6)、单纯EPO组(EPO,n=18)、急性肾损伤组(AKI,n=18)和EPO干预组(AKI+EPO,n=18),EPO组、AKI组和AKI+EPO组又分为3个亚组,即1,6和24 h组(均为n=6)。在各自的时间点留取标本,检测血中尿素氮、肌酐和肌红蛋白水平;HE染色法观察肾脏病理;免疫组化观察GRP78和CHOP蛋白表达,实时荧光定量PCR检测GRP78和CHOP mRNA的表达。结果与对照组比较,AKI和AKI+EPO组大鼠尿素氮、肌酐和肌红蛋白水平升高,GRP78和CHOP蛋白及mRNA表达水平均显著上调(P0.05);AKI组肾脏结构出现损伤;与AKI组比较,AKI+EPO组6和24 h血肌酐水平,GRP78、CHOP蛋白和mRNA表达水平较同期均下降(P0.05)。结论 EPO可以通过影响横纹肌溶解致大鼠急性肾损伤时内质网应激相关蛋白的表达,发挥肾脏保护作用,其机制可能与调节未折叠蛋白反应有关。     

Gamma/deltaT-cell subsets, NKG2A expression and apoptosis of Vdelta2+ T cells in pregnant women with or without risk of premature pregnancy termination

PROBLEM: Potentially cytotoxic Vdelta2+ T lymphocytes recognize human leukocyte antigen-E on the trophoblast via their CD94/NKG2A receptors. This study aims at determing the percentage of gamma/delta T-cell subsets, their NKG2A and Annexin V positivity in peripheral blood of healthy pregnant women and women at risk of premature pregnancy termination. METHOD OF STUDY: Peripheral Vdelta2+ cells from healthy pregnant women and from women at risk of premature pregnancy termination were tested for the KIR NKG2A and Annexin V positivity by flow cytometry. RESULTS: The percentage of viable Vdelta2+ T cells was higher, that of Vdelta1+ T cells was lower in women at risk of premature pregnancy termination than in healthy pregnant women. The percentage of NKG2A + Vdelta2+ T cells was significantly lower in pregnant women at risk of premature pregnancy termination than in normal pregnancy. CONCLUSIONS: These data suggest the involvement of gamma/delta T lymphocytes in the pathogenesis of premature pregnancy termination.     

CRIPTO promotes an aggressive tumour phenotype and resistance to treatment in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is the third leading cause of cancer‐related death worldwide. Despite increasing treatment options for this disease, prognosis remains poor. CRIPTO (TDGF1) protein is expressed at high levels in several human tumours and promotes oncogenic phenotype. Its expression has been correlated to poor prognosis in HCC. In this study, we aimed to elucidate the basis for the effects of CRIPTO in HCC. We investigated CRIPTO expression levels in three cohorts of clinical cirrhotic and HCC specimens. We addressed the role of CRIPTO in hepatic tumourigenesis using Cre‐loxP‐controlled lentiviral vectors expressing CRIPTO in cell line‐derived xenografts. Responses to standard treatments (sorafenib, doxorubicin) were assessed directly on xenograft‐derived ex vivo tumour slices. CRIPTO‐overexpressing patient‐derived xenografts were established and used for ex vivo drug response assays. The effects of sorafenib and doxorubicin treatment in combination with a CRIPTO pathway inhibitor were tested in ex vivo cultures of xenograft models and 3D cultures. CRIPTO protein was found highly expressed in human cirrhosis and hepatocellular carcinoma specimens but not in those of healthy participants. Stable overexpression of CRIPTO in human HepG2 cells caused epithelial‐to‐mesenchymal transition, increased expression of cancer stem cell markers, and enhanced cell proliferation and migration. HepG2‐CRIPTO cells formed tumours when injected into immune‐compromised mice, whereas HepG2 cells lacking stable CRIPTO overexpression did not. High‐level CRIPTO expression in xenograft models was associated with resistance to sorafenib, which could be modulated using a CRIPTO pathway inhibitor in ex vivo tumour slices. Our data suggest that a subgroup of CRIPTO‐expressing HCC patients may benefit from a combinatorial treatment scheme and that sorafenib resistance may be circumvented by inhibition of the CRIPTO pathway. Copyright © 2018 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

内质网应激诱导人乳腺癌MDA-MB-231细胞系凋亡

目的探讨内质网应激(ERS)致雌激素受体阴性(ER-)人乳腺癌MDA-MB-231细胞系凋亡及钙激活中性蛋白酶2(calpain2)在凋亡效应中的作用。方法实验设空白对照组(DMEM)、对照组(DMEM+DMSO)和实验组,用不同浓度衣霉素(TM)诱导乳腺癌细胞不同时间,MTT法和流式细胞术检测细胞增殖抑制率及凋亡率;Western blot法检测葡萄糖调节蛋白78(GRP78)表达量,以GRP78表达最高点确定ERS模型建立,检测凋亡相关蛋白caspase4和CHOP表达以及calpain及其内源性抑制酶calpastatin的表达,用ERS抑制剂(4-PBA)和calpain抑制剂(calpeptin)分别预处理细胞2 h,观察对上述效应的影响。结果 9、12和18μmol/L TM诱导ERS对该细胞增殖有明显抑制效应,抑制率分别为33.88%±1.32%、51.51%±8.85%和55.77%±2.61%,细胞凋亡率分别为9μmol/L TM组16.70%±0.46%和12μmol/L TM组28.1%±1.09%,与对照组相比有明显差异(P0.05);9μmol/L TM诱导细胞24 h的GRP78表达上调最高(P0.01);ERS还可明显上调caspase4、CHOP和calpain表达,下调calpastatin表达(P0.01或P0.05),上述效应均能被4-PBA和calpeptin减弱或阻断(P0.05)。结论 ERS可诱导MDA-MB-231细胞凋亡,calpain2参与调控凋亡发生。