乙型病毒性肝炎肝硬化门静脉高压症患者脾切除后CD4＋CD25＋CD127low/-调节性T细胞的变化及意义

目的 观察乙型病毒性肝炎肝硬化门静脉高压症患者行脾切除术前后外周血中CD4＋CD25＋ CD127low/-调节性T细胞（Treg）的变化,探讨门静脉高压症患者行脾切除术对机体免疫功能的影响.方法 回顾性分析2012年5月至2013年5月河北医科大学第三医院收治的20例乙型病毒性肝炎肝硬化门静脉高压症合并脾功能亢进患者的临床资料,通过流式细胞仪分析乙型病毒性肝炎肝硬化门静脉高压症合并脾功能亢进患者（门静脉高压症组）脾切除术前1d、术后1周、1个月、3个月和来自河北医科大学第三医院的10例健康体检者（对照组）的外周血中CD4＋ CD25＋ CD127low/-Treg含量,分析Treg的变化对全身免疫系统造成的影响.组问比较采用t检验,手术前后数据比较采用重复测量方差分析.结果 门静脉高压症组患者术前和对照组受试者CD4＋ CD25＋ CD127low/-Treg比例分别为5.1％±3.5％和1.4％±0.2％,两组比较,差异有统计学意义（t=2.573,P＜0.05）.门静脉高压症组患者术后1周、1个月、3个月CD4＋CD25＋ CD127low/-Treg比例分别为9.2％±2.7％、5.6％±1.7％、2.5％±2.1％,其中术后1周与术前比较,差异有统计学意义（F=9.814,P＜0.05）;而术后3个月与术前比较,差异无统计学意义（F=2.364,P＞0.05）.结论 脾切除术后短期Treg水平明显升高,随时间延长逐渐下降.从Treg方面表明脾切除对机体免疫系统影响较小.     

不同穿刺角度对超声引导下经皮肾活检取材质量和安全性的影响

目的 比较垂直进针和45°斜向下进针时经皮肾活检的取材质量和安全性.方法 回顾成都市核工业四一六医院肾内科2003年4月至2012年4月203例超声引导下经皮肾活检患者的临床资料,根据进针方向将患者分为2组：垂直进针组（A组）79例,45°斜向下进针组（B组）124例.比较分析2组患者取材质量（单次取材长度、切片包含肾小球数均值和取中皮髓交界处患者比例）和安全性（皮下血肿发生率、镜下血尿发生率、血红蛋白下降患者比例）.结果 两组单次取材长度的差异无统计学意义[（0.89±0.09）cm比（1.08±0.05）cm,P＞0.05].B组切片中肾小球数[（24±4）个比（15±6）个,P＜0.05]及取中皮髓交界处患者比例多于A组[67.7％（84/124）比17.7％ （14/79）,P＜0.01].B组镜下血尿发生率[23.4％ （29/124）比37.9％ （30/79）]和血红蛋白下降比例[4.8％ （6/124）比15.2％ （12/79）]均低于A组（均P＜0.05）,两组皮下血肿的发生率差异无统计学意义（P＞0.05）.结论 45°斜向下进针不仅可提高肾活检的取材质量,还在一定程度上减少了其并发症的发生率.     

阑尾黏液性肿瘤病理学特征及预后分析

目的探讨基于2010版WHO分类的各类阑尾黏液性肿瘤的病理学特征及预后。方法收集并复习2003年1月至2012年6月间在南京大学医学院附属鼓楼医院接受手术切除的70例阑尾黏液性肿瘤术后病理切片,按照2010年消化系统肿瘤WHO新分类,将阑尾黏液性肿瘤分为5种类型：阑尾黏液性腺瘤／囊腺瘤（MA）、低级别黏液性肿瘤（LAMN）、起源于阑尾的低级别腹膜假黏液瘤（PMP．L）、浸润性黏液腺癌（MAC）以及起源于阑尾的高级别腹膜假黏液瘤（PMP．H）。结果11例MA肿瘤上皮和黏液仅局限于黏膜肌层以内;术后无复发或死亡病例。41例LAMN黏液均可出现于黏膜肌层外阑尾壁或阑尾表面,其中39黏液湖内没有或仅见少数黏液上皮,上皮具有轻度异型性;有3例复发或进展,元死亡病例。7例PMP—L上皮具有轻度异型性,其中有4例黏液湖内上皮数量稀少或缺乏,其余3例上皮数量中等至较多：术后复发和死亡各1例。7例MAC和4例PMP—H均为浸润性肿瘤,至少局部出现高级别细胞学特征;术后复发4例,死亡3例（含2例复发后死亡）。MA和LAMN在组织学上表现为非浸润性肿瘤。而PMP-L、MAC和PMP-H则在生物学行为上表现为腺癌特征。结论阑尾黏液性肿瘤是一系列肿瘤谱系,其生物学行为取决于肿瘤性质和类型,统一、规范而准确的病理诊断对于治疗至关重要。     

乳晕缩小成形术两种切口的效果比较

目的比较两种不同方法行乳晕缩小成形术的效果。方法对37例女性行乳晕缩小成形术,所有受术者随机分为2组：A组19例,采用常规乳晕双环形切口;B组18例,采用经乳头根部乳晕双环形切口。比较二者手术时间、手术瘢痕、术后皮肤皱褶吸收时间及乳头乳晕感觉的差异。结果A、B组平均手术时间分别为40．5min及45．8min,差异有统计学意义（P〈0．05）。2组乳头乳晕感觉无差异,2组均无围术期并发症发生,2组“荷包”皮肤缝合皱褶吸收时间分别为4．34个月和8．78个月,差异有统计学意义（P％0．01）。B组较A组瘢痕更加隐蔽,患者满意度更高。结论与常规乳晕双环形切口相比,经乳头根部双环形切口术后瘢痕更为隐蔽,但要皮肤皱褶吸收时间更长。     

The ryanodine receptor agonist 4-chloro-3-ethylphenol blocks ORAI store-operated channels

BACKGROUND

Depletion of the Ca²⁺ store by ryanodine receptor (RyR) agonists induces store-operated Ca²⁺ entry (SOCE). 4-Chloro-3-ethylphenol (4-CEP) and 4-chloro-m-cresol (4-CmC) are RyR agonists commonly used as research tools and diagnostic reagents for malignant hyperthermia. Here, we investigated the effects of 4-CEP and its analogues on SOCE.

EXPERIMENTAL APPROACH

SOCE and ORAI1-3 currents were recorded by Ca²⁺ imaging and whole-cell patch recordings in rat L6 myoblasts and in HEK293 cells overexpressing STIM1/ORAI1-3.

KEY RESULTS

4-CEP induced a significant release of Ca²⁺ in rat L6 myoblasts, but inhibited SOCE. The inhibitory effect was concentration-dependent and more potent than its analogues 4-CmC and 4-chlorophenol (4-ClP). In the HEK293 T-REx cells overexpressing STIM1/ORAI1-3, 4-CEP inhibited the ORAI1, ORAI2 and ORAI3 currents evoked by thapsigargin. The 2-APB-induced ORAI3 current was also blocked by 4-CEP. This inhibitory effect was reversible and independent of the Ca²⁺ release. The two analogues, 4-CmC and 4-ClP, also inhibited the ORAI1-3 channels. Excised patch and intracellular application of 4-CEP demonstrated that the action site was located extracellularly. Moreover, 4-CEP evoked STIM1 translocation and subplasmalemmal clustering through its Ca²⁺ store-depleting effect via the activation of RyR, but no effect on STIM1 redistribution was observed in cells co-expressing STIM1/ORAI1-3.

CONCLUSION AND IMPLICATIONS

4-CEP not only acts as a RyR agonist to deplete the Ca²⁺ store and trigger STIM1 subplasmalemmal translocation and clustering, but also directly inhibits ORAI1-3 channels. These findings demonstrate a novel pharmacological property for the chlorophenol derivatives that act as RyR agonists.     

Synthesis of a stable isotopically labeled universal surrogate peptide for use as an internal standard in LC‐MS/MS bioanalysis of human IgG and Fc‐fusion protein drug candidates

The synthesis of a 16‐residue, stable isotopically labeled peptide is described for use as a LC‐MS/MS (Liquid chromatography‐mass spectrometry/mass spectrometry) internal standard in bioanalytical studies. This peptide serves as a single universal surrogate peptide capable of quantifying a wide variety of immunoglobulin G and Fc‐fusion protein drug candidates in animal species used in pre‐clinical drug development studies. An efficient synthesis approach for this peptide was developed using microwave‐assisted solid phase peptide synthesis (SPPS) techniques, which included the use of a pseudoproline dipeptide derivative. The corresponding conventional room temperature SPPS was unsuccessful and gave only mixtures of truncated products. Stable‐labeled leucine was incorporated as a single residue via manual coupling of commercially available Fmoc‐[¹³C₆, ¹⁵N]‐l ‐leucine onto an 11‐unit segment followed by automated microwave‐assisted elaboration of the final four residues. Using this approach, the desired labeled peptide was prepared in high purity and in sufficient quantities for long‐term supplies as a bioanalytical internal standard. The results strongly demonstrate the importance of utilizing both microwave‐assisted peptide synthesis and pseudoproline dipeptide techniques to allow the preparation of labeled peptides with highly lipophilic and sterically hindered side‐chains.     

A novel gastroretentive porous microparticle for anti-Helicobacter pylori therapy: Preparation, in vitro and in vivo evaluation

Gastroretentive drug delivery system is a promising option for the treatment of Helicobacter pylori infection, which can prolong gastric residence time and supply high drug concentration in the stomach. In the present study, a low density system of metronidazole-loaded porous Eudragit^® RS microparticle with high drug loading capacity (>25%) was fabricated via electrospray method. The porous structure and size distribution of microparticles were affected by polymer concentration and flow rate of solution. FTIR and XRD analyses indicated that drug has been entrapped into the porous microparticles. In addition, sustained release profiles and slight cytotoxicity in vitro were detected. Gamma scintigraphy study in vivo demonstrated that ¹³¹I-labeled microparticles retained in stomach for over 8 h, and about 65.50% radioactive counts were finally detected in the region of interest. The biodistribution study confirmed that hotspot of radioactivity was remaining in the stomach. Furthermore, metronidazole-loaded porous microparticles can eradicate H. pylori completely with lower dose and administration frequency of antibiotic compared with pure drug, which were also more helpful for the healing of mucosal damages. These results suggest that prepared porous microparticle has the potential to provide better treatment for H. pylori infection.     

Perfluorooctanoic acid stimulates breast cancer cells invasion and up-regulates matrix metalloproteinase-2/-9 expression mediated by activating NF-κB

Perfluorooctanoic acid (PFOA) is widely used because of its stain-resistant and water-repellant properties. This study aimed to explore the molecular mechanisms undergoing the stimulation effects of PFOA on cancer cell invasion and matrix metalloproteinases (MMPs) expression. Trans-well filter assay showed that PFOA exposure (≥5 nM) evidently enhanced the invasion ability of the breast cancer cells MDA-MB-231. Luciferase reporter assay, quantitative real-time PCR, western blotting and gelatin zymography consistently demonstrated that mRNA and protein levels of MMP-2/-9 were increased in the cells after PFOA treatment (P < 0.05 each). Western blotting revealed that PFOA could activate nuclear factor kappaB (NF-κB) by accelerating NF-κB translocation into the nucleus. Furthermore, addition of NF-κB inhibitor in culture medium could suppress the breast cancer cells invasiveness enhancement and MMP-2/-9 overexpression. This study indicates that PFOA can stimulate breast cancer cells invasion and up-regulate matrix metalloproteinase-2/-9 expression mediated by activating NF-κB, which deserves more environmental health concerns.     

全肠外营养在术后恶性肿瘤患者治疗中的应用与进展

肿瘤患者大多术前已存在不同程度的营养不良及免疫功能障碍,而手术的创伤和术后应激反应引起的分解代谢加强更加重了营养不良和免疫抑制,导致机体抵抗力降低,并发症增多,死亡率增高和住院时间延长。全肠外营养（ TPN）支持可增加肿瘤患者特别是术后恶性肿瘤患者肌肉组织和肝脏内糖原的贮存,使不能正常进食或超高代谢肿瘤患者维持较好的营养状态,增强自身免疫能力,提高其生存质量。此外,很多术后恶性肿瘤患者在术后营养不良的情况下,还在多次进行放、化疗,不但机体耐受能力下降,明显影响化疗效果,还严重影响患者生活及生存质量。因此,关注术后恶性肿瘤患者营养支持需求,特别是营养的平衡供给尤为重要。本文就TPN在术后恶性肿瘤的应用状况与研究进展做一综述。