非程控降温,—80℃冻存自体外周血干细胞的研究

目的：观察非程控降温、－80℃冻存的方法对自体外周血十细胞（APBSC）的保存效果。方法：以6％羟乙基淀粉（HES）、5％二甲基亚砜（DMSO）及4％人血白蛋白（ALB）的混合物为冷冻防护剂，将APBSC直接置于－80℃下保存，冻存前反复苏后测定APBSC的CFU－GM、BFU－E；观察移植后造血功能重建情况。结果：13例患者白细胞在十3～＋7天下降至（0．0～0．1）×10／L，白细胞（0．0～0．2）×109／L持续时间3～6天，于＋9～＋11天恢复至1．0X109／L以上．中性粒细胞绝对值（ANC）于＋9～＋11天达到0．5X109／L。血小板在＋3～＋7天下降至（2．0～21）×109／L，于＋8～＋15天恢复至20×109／L以上。CFU－GM、BFU－E回大率分别为76．5％、78．4％。结论：非程控降温、－80℃冻存是一简便、经济、有效的自体外周血于细胞保存方法。     

Isolation of pathogenic bacteria from hospital staff apparel in Nigeria

A survey of bacteria contamination of hospital staff apparel in use in Anambra State, Nigeria, was carried out to determine the extent of contamination by clinically important bacteria. Of a total of 125 swab samples of hospital staff apparel, 72 (58%) showed bacterial contamination including 32 (70%) of 46 samples from hand gloves, 28 of 45 (62%) samples from protective gowns, and 12 of 34 (35%) samples from face-shields. The potentially pathogenic bacteria isolated were Salmonella spp, Proteus vulgaris, Shigella dysenteriae, Pseudomonas aeruginosa and Staphylococcus aureus. The isolation of clinically important bacteria from the apparel suggests the need for improved infection control measures.     

大鼠急性肺栓塞后SMP-30表达下降及其对细胞凋亡的影响

目的：研究大鼠急性肺栓塞模型肺组织中衰老标记蛋白质30(SMP-30)的表达变化及其对Fas诱导的细胞凋亡的影响。方法:建立大鼠急性肺栓塞模型，分别在急性肺栓塞后1、8、24和48 h进行支气管肺泡灌洗，然后开胸取出肺组织。常规提取肺组织的总RNA和总蛋白，以正常组为对照，采取半定量RT-PCR的方法研究SMP-30在mRNA水平表达的变化；采用Western blotting方法进一步验证SMP-30在蛋白水平表达的变化；采用免疫组织化学方法检测大鼠肺组织中SMP-30以及肺泡巨噬细胞中IL-8在肺栓塞前后表达的变化及其组织分布情况；采用TUNEL法研究急性肺栓塞后组织细胞的凋亡情况；最后采用ELISA法检测急性肺栓塞后肺泡灌洗液中sFasL的浓度变化。结果:在大鼠急性肺栓塞后的不同时点，SMP-30的mRNA水平和蛋白水平均逐渐降低，在24和48 h下降最为明显。免疫组化研究表明SMP-30主要分布在支气管黏膜上皮细胞和肺泡上皮细胞，急性肺栓塞后SMP-30在上述细胞内的表达均明显降低。TUNEL染色发现随着SMP-30表达的降低，肺组织内出现明显的细胞凋亡现象，同时肺泡灌洗液中sFasL的浓度升高，肺泡巨噬细胞内IL-8的表达也明显升高。结论:大鼠急性肺栓塞后肺组织内SMP-30的表达明显降低，可能促进Fas－FasL细胞凋亡系统的活化。     

白细胞介素4剪接变异体在哮喘患者中的表达

目的检测哮喘患者外周血中IL-4及其变异体IL-4δ2的表达,探讨其在哮喘发病机制中的作用。方法采用半定量RT-PCR技术,检测10例哮喘患者IL-4及其变异体的表达,并与正常健康人群进行比较。结果哮喘患者IL-4表达较健康人群高,而且IL-4和IL-4δ2的比值远远高于健康人。结论IL-4和IL-4δ2表达的相对性可能在哮喘发病机制中具有重要作用,其可能是Th2细胞在不同临床疾病中功能多样性的原因之一。     

联合门静脉／肠系膜上静脉切除的胰头癌根治术

目的探讨胰头癌侵犯门静脉（portalvein，PV）和（／或）肠系膜上静脉（superior mesentericvein，SMV）时根治切除的可行性。方法回顾分析11例PV／SMV受侵的胰头癌患者临床资料，均行扩大胰十二指肠切除术。其中7例行血管壁部分切除，3例行血管节段性切除及对端吻合，1例行受侵血管切除＋人工血管移植。脾静脉与SMV端侧吻合4例，脾静脉结扎3例。消化道重建采用Child术式。结果本组PV阻断时间平均为18．1（9～32）min。全组患者术后均未发生血管栓塞、肠坏死、肝衰竭等并发症，均康复出院。11例均获随访，时间6～20个月，3例术后1年内死亡，4例术后1—2年死亡，患者平均生存时间15（7～20）个月。结论对单纯侵犯PV／SMV的胰头癌施行联合PV／SMV切除的胰头癌扩大根治术是安全可行的。     

Fragile X‐associated tremor/ataxia syndrome (FXTAS) in grey zone carriers

The grey zone (GZ; 45–54 CGG repeats in the FMR1 gene) is considered a normal allele; however, several studies have found a high frequency of GZ in movement disordered populations. Here, we describe neurological features of fragile X‐associated tremor/ataxia syndrome (FXTAS) in two carriers of GZ alleles, although FXTAS has been defined as occurring only in premutation carriers (55–200 CGG repeats). Both patients had family members who had premutation and were diagnosed with FXTAS. The presence of relatively high GZ alleles with elevated fragile X mental retardation 1 mRNA (FMR1‐mRNA) combined with a family history of FXTAS that may represent a facilitating genetic background for FXTAS are the factors that led to the presence of FXTAS in these individuals with a GZ allele. Further research into clinical involvement of GZ alleles is recommended and the definition of FXTAS may require revision.     

Macrophages retain hematopoietic stem cells in the spleen via VCAM-1

Splenic myelopoiesis provides a steady flow of leukocytes to inflamed tissues, and leukocytosis correlates with cardiovascular mortality. Yet regulation of hematopoietic stem cell (HSC) activity in the spleen is incompletely understood. Here, we show that red pulp vascular cell adhesion molecule 1 (VCAM-1)⁺ macrophages are essential to extramedullary myelopoiesis because these macrophages use the adhesion molecule VCAM-1 to retain HSCs in the spleen. Nanoparticle-enabled in vivo RNAi silencing of the receptor for macrophage colony stimulation factor (M-CSFR) blocked splenic macrophage maturation, reduced splenic VCAM-1 expression and compromised splenic HSC retention. Both, depleting macrophages in CD169 iDTR mice or silencing VCAM-1 in macrophages released HSCs from the spleen. When we silenced either VCAM-1 or M-CSFR in mice with myocardial infarction or in ApoE^−/− mice with atherosclerosis, nanoparticle-enabled in vivo RNAi mitigated blood leukocytosis, limited inflammation in the ischemic heart, and reduced myeloid cell numbers in atherosclerotic plaques.Leukocytosis correlates closely with cardiovascular mortality. In the steady state, blood leukocytes derive exclusively from bone marrow hematopoietic stem cells (HSCs). Supporting cells (Sugiyama et al., 2006; Ding et al., 2012; Ding and Morrison, 2013), including macrophages (Winkler et al., 2010; Chow et al., 2011), maintain the bone marrow HSC niche and regulate hematopoietic stem and progenitor cell (HSPC) activity by supplying various cytokines and retention factors. Systemic inflammation can stimulate extramedullary hematopoiesis in adult mice and humans. Splenic myelopoiesis supplies inflammatory monocytes to atherosclerotic plaques (Robbins et al., 2012) and the ischemic myocardium (Leuschner et al., 2012). In ischemic heart disease, HSPCs emigrate from the bone marrow, seed the spleen, and amplify leukocyte production (Dutta et al., 2012). Splenic HSPCs localize in the red pulp near the sinusoids in parafollicular areas (Kiel et al., 2005). Likewise, after adoptive transfer of GFP⁺ HSPCs, GFP⁺ colonies populate the splenic red pulp of atherosclerotic ApoE^−/− mice (Robbins et al., 2012). During myocardial infarction (MI), proinflammatory monocytes derived from the spleen accelerate atherosclerotic progression (Dutta et al., 2012). Collectively, these data suggest that splenic myelopoiesis has promise as a therapeutic target; however, the components of the splenic hematopoietic niche are incompletely understood, especially compared with the well-studied bone marrow niche. Understanding HSC retention factors and their regulation in the spleen was the purpose of this study.Because the spleen harbors very few HSCs in the steady state, we investigated the splenic hematopoietic niche after injecting the Toll-like receptor ligand LPS to activate extramedullary hematopoiesis. In the bone marrow, macrophages are an integral part of the HSC niche (Winkler et al., 2010; Chow et al., 2011) and differentiation depends on the receptor for macrophage colony-stimulating factor (M-CSFR, CD115; Auffray et al., 2009). We thus hypothesized that splenic hematopoietic niche assembly also requires M-CSFR signaling. In line with knockout studies (Takahashi et al., 1994; Dai et al., 2002), in vivo knockdown of M-CSFR with nanoparticle-encapsulated siRNA reduced splenic macrophage numbers substantially. Interestingly, decreased macrophage numbers were associated with a reduction of splenic HSCs. Depleting macrophages with diphtheria toxin (DT) in CD169 iDTR mice reproduced the findings obtained with M-CSF–directed siRNA treatment, thereby indicating that macrophages have a key role in splenic HSC maintenance. To investigate how splenic macrophages retain HSCs, we measured changes in splenic expression of major bone marrow retention factors after M-CSFR silencing. Silencing M-CSFR selectively reduced splenic VCAM-1, and the adhesion molecule was primarily expressed by macrophages. Inhibiting macrophage expression of VCAM-1 with siRNA targeting this adhesion molecule reduced splenic HSPC numbers. Finally, we found that M-CSFR and macrophage-directed VCAM-1 silencing in mice with atherosclerosis mitigated blood leukocytosis and dampened inflammation in atherosclerotic plaques and the infarcted myocardium. These data reveal the importance of VCAM-1 expression by splenic macrophages for extramedullary hematopoiesis and illustrate the therapeutic potential of RNAi as an antiinflammatory that mutes emergency overproduction and provision of myeloid cells.     

参麦注射液联合氢化可的松治疗老年脓毒性休克的临床研究

目的探索参麦注射液联合氢化可的松对老年脓毒性休克的治疗效果。方法选取2015年7月—2018年7月沧州市人民医院收治的脓毒性休克患者102例,随机分为对照组(51例)和治疗组(51例)。对照组静脉滴注注射用氢化可的松琥珀酸钠,200 mg加入生理盐水250 mL,1次/d;治疗组在对照组基础上静脉滴注参麦注射液,100 mL/次,1次/d。两组患者均持续治疗5天。观察两组患者临床疗效,同时比较治疗前后两组患者白细胞计数、APACHEⅡ评分、乳酸、降钙素原和脑钠肽水平及随访结果。结果治疗后,对照组和治疗组临床有效率分别为76.47%和86.27%,两组比较差异具有统计学意义(P0.05)。治疗后,两组患者白细胞计数、C-反应蛋白水平及APACHEⅡ评分均明显降低(P0.05),且治疗组白细胞计数、C-反应蛋白水平及APACHEⅡ评分明显低于对照组(P0.05)。治疗后,两组患者乳酸、降钙素原和脑钠肽水平均显著降低(P0.05),且治疗组乳酸、降钙素原和脑钠肽水平明显低于对照组(P0.05)。治疗组ICU住院时间、14 d死亡率及28 d死亡率上均优于对照组,两组比较差异具有统计学意义(P0.05)。结论参麦注射液联合氢化可的松可明显改善老年脓毒性休克患者早期生理生化指标,提高整体生存率。     

大鼠急性肺栓塞模型中CA3和NHE1表达的变化

目的研究大鼠急性肺栓塞模型肺组织中与酸碱平衡相关的碳酸酐酶3（CA3）和Na+/H+交换器（NHE1）的表达变化。方法建立大鼠急性肺栓塞模型,分别在急性肺栓塞后1、8、24和48 h开胸取出肺组织,然后提取肺组织的总RNA和总蛋白,以正常组为对照,采取半定量RT-PCR的方法研究CA3和NHE1在mRNA水平表达的变化;采用western-b lot方法进一步验证CA3和NHE1在蛋白水平表达的变化;同时采用免疫组织化学的方法检测大鼠肺组织中NHE1在肺栓塞前后表达的变化。结果在大鼠急性肺栓塞后的不同时间点,CA3的mRNA水平和蛋白水平均逐渐降低,而NHE1在mRNA水平和蛋白水平的表达都出现逐渐升高现象。免疫组化研究表明NHE1主要分布在支气管黏膜上皮和细支气管黏膜上皮的胞浆内,急性肺栓塞后NHE1在上皮细胞内的表达明显升高。结论大鼠急性肺栓塞后肺组织内CA3的表达降低,NHE1的表达升高,这一现象可能与机体的酸碱平衡调节相关。