经静脉移植骨髓间充质干细胞向心肌梗死区趋化的组织学观察

目的 采用2-脱氧5氮杂胞苷（5-aza-2’-deoxycytidine，5-aza-CdR）诱导的骨髓间充质干细胞（MSCs）经尾静脉移植入不同时期的心肌梗死（MI）后心衰大鼠中,观察其向病损心肌的趋化作用及其在局部分布的特点，探讨其对心肌α-肌球蛋白重链（α-MHC）mRNA表达的影响。方法 培养Wistar大鼠的MSCs，用0.3 μmol/L的5-aza-CdR两次诱导第2代的MSCs。用溴氮胞苷（BrdU）标记后，经尾静脉植入MI后不同时期的心衰大鼠中，实验分为MI 1 d、8 d、3周、1月后移植组及MI 3周、1月后DMEM假移植组。采用免疫组化染色法检测移植后3 d及1月的MSCs在大鼠心肌中的分布情况，并用RT-PCR检测移植1月后MI局部α-MHC mRNA水平的变化。结果 在MI后不同时期（1 d内、8 d、3周、1月），通过尾静脉给予相同数量的MSCs，3 d后在各组大鼠的心肌中均发现抗BrdU＋的移植细胞。移植的MSCs主要分布在心肌受损区域，分布特点与移植时心肌的病理特点有一定的关系。急性、亚急性期，有较多的移植细胞，与自身细胞均匀分布。进入慢性期，移植细胞沿着纤维方向线样排列，呈团簇样分布，数目较急性期减少。移植后1月，各组大鼠心脏中仍有抗BrdU+的移植细胞，移植细胞分布的特点与移植后3 d所观察的结果相似。研究发现，MI区域rattus α-MHC mRNA的表达，与MI 3周移植的MSCs相比，在MI 1 d或8 d后移植，差异无显著性。MI 3周后移植MSCs，rattus α-MHC mRNA的表达较DMEM假移植组有明显提高(P<0.01)，但在MI 1月后移植与假移植组差异无显著性。结论 在不同的MI时间段经尾静脉植入MSCs，其能向心肌趋化，主要分布在心肌的受损区域，分布特点与移植时心肌的病理特点有关。MI后早期给予MSCs移植，更有利于提高MI区域rattus α-MHC mRNA的表达。     

骨髓间充质干细胞在心肌梗死区的移植和分化对心肌梗死后心力衰竭的影响

目的采用2′脱氧5-氮杂胞苷（5-aza-CdR）诱导的骨髓间充质干细胞（MSCs）移植于心肌梗死后心力衰竭大鼠心肌中,评价其存活、分化及对心功能的影响。方法将经5-aza-CdR（0.3μmol/L）两次诱导,溴氮胞苷标记的第2代大鼠MSCs植入心肌梗死后10 d心力衰竭大鼠心肌疤痕中,同时以植入无血清培养基（DMEM）的大鼠为对照。移植前及移植后1月,通过检测左室射血分数（LVEF）等超声指标研究其对心功能的影响;应用双重免疫组化及电镜观察研究MSCs在心肌梗死区的存活和分化。结果移植后1月,MSCs组大鼠心功能[LVEF=（83±8）%,n=13]较DMEM组[LVEF=（47±12）%,n=12]显著改善（P<0.01）;并且较移植前[LVEF=（64±10）%,n=13]也有显著提高（P<0.01）。移植的MSCs能在心肌及疤痕中存活,并向心肌细胞分化,Tropon inT及Connex-in43阳性。MSCs有肌丝形成,并与周围心肌细胞紧密连接,类似闰盘。结论经5-aza-CdR诱导的MSCs移植后1月,能在心肌梗死后心力衰竭大鼠心肌及疤痕中存活并向心肌细胞分化,且可改善大鼠的心功能。     

兔骨髓间充质干细胞移植后移植细胞凋亡的实验研究

目的利用TUNEL法检测5-氮胞苷体外诱导骨髓间充质干细胞（MSCs）移植后的凋亡情况。方法5-氮胞苷体外诱导骨髓间充质干细胞MSCs向肌源性心肌细胞分化,通过免疫组化,鉴定诱导后的MSCs是否向类心肌细胞转化。建立兔心肌梗死模型,将细胞移植于心梗区域。移值2周后,利用TUNEL法检测植入细胞的凋亡率。结果移植2周后,可见DAPI标记带蓝色荧光的供体细胞核,分布比较广泛,形态呈椭圆形类似心肌细胞核,并与心肌纤维排列方向一致,证明移植细胞已存活。移值细胞表达troponinT,证明移植的MSCs分化为类心肌细胞。移植细胞均出现不同程度细胞凋亡。结论移植的MSCs细胞可在缺血的心肌组织存活,并分化为类心肌细胞,但移植细胞均出现不同程度凋亡。     

异体骨髓基质干细胞移植治疗慢性心力衰竭大鼠的疗效

目的分离、培养、诱导大鼠骨髓基质干细胞(MSCs),并将其移植入慢性心力衰竭(CHF)大鼠并检测其治疗效果,探讨异体MSCs修复CHF受损心肌的能力。方法注射法分离幼龄大鼠胫骨中的MSCs并使用5-氮胞苷对其进行诱导分化。使用阿霉素鼠尾静脉注射法制备大鼠CHF模型,将大鼠分为4组,分别为正常非移植组10只,正常移植组10只,CHF非移植组20只,CHF移植组20只。将诱导分化的MSCs手术注射入左心室部位,对照组植入等量培养液。术后4 w进行超声检测及血流动力学分析,然后取心脏行免疫组织化学分析。结果 5-氮胞苷诱导后MSCs可分化为心肌样细胞。将其移植入CHF大鼠后,超声及血流动力学结果显示,与CHF非移植组比较,移植干细胞组左室收缩末压、左室射血分数和左室压力变化速率最大值均显著升高(P<0.05,P<0.01),左室舒张末压显著降低(P<0.05);术后4 wCHF移植组可以检测到Brdu标记阳性细胞。结论 CHF后移植MSCs可改善心功能,提高左室射血分数,为慢性心衰的治疗开辟了新的治疗途径。     

诱导对骨髓间充质干细胞移植后凋亡影响的实验研究

目的 研究兔骨髓间充质干细胞(MSCs)诱导与移植细胞凋亡的关系.方法 无菌条件下采集兔MSCs,一部分不诱导,一部分经5-氮胞苷诱导使其向类心肌细胞分化;将干细胞移植于兔心肌梗死区.实验动物随机分两组:MSCs组(n=20)心梗后1 h在心梗区域注入MSCs;经诱导组(n=20)心梗后1 h在心梗区域注入经诱导的MSCs.2周后用荧光显微镜观察移值细胞是否存活,利用TUNEL法检测植入细胞的凋亡率.结果 MSCs经5-氮胞苷诱导,传代培养2～4周,细胞表达肌钙蛋白T(trponin T),证明MSCs向心肌细胞转化.移植2周后,用荧光显微镜观察,在梗死区组织标本中可见DAPI标记带蓝色荧光的供体细胞核,形态呈椭圆形类似心肌细胞核,并与心肌纤维排列方向一致,证明移植细胞已存活.利用TUNEL法检测,两组移植细胞均出现细胞凋亡,差异无统计学意义.结论 MSCs移植入心肌组织后的凋亡与5-氮胞苷诱导无关.     

骨髓间充质干细胞移植在心肌病大鼠体内存活、分化的实验研究

目的:研究骨髓间充质干细胞（MSCs）移植在心肌病大鼠体内存活、分化的情况。方法:将雌性W istar大鼠30只经腹腔注射阿霉素6次（总剂量15mg/kg）,建立心肌病的动物模型,在此模型的左室前壁采用微静脉注射法植入MSCs。8周后处死大鼠,取出心脏标本,做冰冻切片进行HE染色,观察植入的MSCs在病损心肌组织中的变化及免疫荧光检查观察植入MSCs心肌肌球蛋白重链（MHC）及心肌特有的连接蛋白（CX43）表达情况。结果:HE染色检查显示,植入的MSCs存活并有新生血管形成;免疫荧光检查显示植入的MSCs,表达MHC及CX43,但较宿主表达弱。结论:MSCs移植在心肌病大鼠体内心肌微环境中能存活,并向心肌细胞分化。     

5-氮胞苷诱导间质干细胞为心肌细胞的实验研究

分离SD大鼠下肢骨，培养获得骨髓间质干细胞（MSCs），经5-氮胞苷（5-aza）定向诱导24h后t用免疫细胞化学法检测诱导细胞对连接蛋白43和α横纹肌肌动蛋白的表达，用逆转录-聚合酶链反应鉴定心肌细胞特异性蛋白的表达情况。结果显示，MSCs经5-aza诱导后，细胞形态不规则；3周后5-aza 10μm组细胞连接蛋白43、α横纹肌肌动蛋白阳性表达；RT-PCR显示细胞表达心肌肌钙蛋白Ⅰ、α心肌肌动蛋白。提示5-aza可体外诱导MSCs分化为心肌细胞，用于心肌梗死的治疗。     

大鼠骨髓间充质干细胞体外定向诱导分化为心肌样细胞的实验研究

目的研究大鼠骨髓间充质干细胞的体外培养及向心肌样细胞转化的条件。方法获取成年大鼠胫骨干骨髓,采用贴壁法进行间充质干细胞的培养、传代,观察经5-氮胞苷诱导后骨髓间充质干细胞的生长和分化。结果大鼠骨髓基质细胞贴壁呈集落生长,5-氮胞苷诱导骨髓基质细胞转化为心肌样细胞。结论骨髓基质细胞能够在体外被诱导分化为心肌样细胞,为自体心肌细胞移植提供了一种良好的来源。     

骨髓间充质干细胞体外定向诱导分化为心肌样细胞的实验研究

目的 研究大鼠骨髓间充质干细胞的体外培养及向心肌样细胞转化的条件。方法 获取成年大鼠胫骨干骨髓，采用贴壁法进行间充质干细胞的培养、传代，观察经5-氮胞苷诱导后骨髓间充质干细胞的生长和分化。结果 大鼠骨髓基质细胞贴壁旱集落生长，5-氮胞苷诱导骨髓基质细胞转化为心肌样细胞，结论 骨髓基质细胞能够在体外被诱导分化为心肌样细胞，为自体心肌细胞移植提供了一种良好的来源。     

骨髓间充质细胞修复心肌梗死的影响因素研究

目的探讨心肌梗死后自体骨髓间充质细胞（MSCs）心肌内移植减轻心功能不全的可能性,以及体外诱导及注射部位等因素对移植疗效的影响。方法34只新西兰白兔均经过骨穿抽取骨髓,开胸建立心梗模型,MSCs体外培养和二次开胸心肌内注射等操作,按照对其MSCs的处理方式和心肌内注射的成分不同随机分为三组：A组为MSCs未经体外诱导;B组为MSCs经5．氮杂胞苷（5-aza）体外诱导;C组为对照组,注射生理盐水。另外根据注射部位不同分为疤痕组和边缘组。结果与未诱导的MSCs相比,5-aza诱导的MSCs移植并未表现出更多的优越性。注射的部位是心功能改善关键的影响因素。如果注射到疤痕区,MSCs更倾向于分化为成纤维细胞。如果注射到边缘区,MSCs则倾向于分化为心肌细胞。结论在进行MSCs心肌内移植时不仅要强调将MSCs投送到梗死区,更要注意将MSCs投送到尚有存活心肌的边缘区。     

胰岛素瘤的解剖分布特点分析

目的胰岛素瘤是最常见的胰腺神经内分泌肿瘤，因其临床表现多样，导致诊断困难。影像学诊断尤其是超声内镜(EUS)在胰岛素瘤的诊断中起着重要作用，拥有较高的敏感性和特异性。本研究拟通过明确胰岛素瘤的解剖分布特点，以期有助于提高影像学的诊断准确率和降低漏诊率，尤其是在教育和培训实践中对于EUS的学习者更具有指导价值。 方法回顾性分析解放军总医院第一医学中心病案资料数据库1993年1月至2019年11月经外科手术、病理确诊为胰岛素瘤的患者的临床资料，检索方法采取搜索术后病理诊断为"胰岛素瘤"的病例，通过查阅病例的方法，提取出胰岛素瘤的大小和解剖分布等数据，进一步分析其特点。 结果共检索到确诊为胰岛素瘤的患者116例，其中，男45例、女71例，年龄13～76岁，平均年龄(44.4±14.85)岁。胰岛素瘤单发110例(94.8%)、多发6例(5.2%)。位置分布：头颈部46例(39.7%)，单发45例、多发1例；体尾部68例(58.6%)，单发65例、多发3例；全胰腺多发2例(1.7%)。病变大小特点：最大径0.4～3.4 cm，平均大小(1.53±0.58)cm。≤1 cm 29例、>1 cm而≤1.5 cm41例、>1.5 cm而≤2.0 cm28例，≤3 cm 15例，>3 cm 3例。年龄与肿瘤的大小相关，≤44岁患者肿瘤平均大小为(1.36±0.51)cm、>44岁患者肿瘤平均大小为(1.70±0.60)cm，P<0.05。头颈部的肿瘤大于体尾部的肿瘤，头颈部肿瘤平均大小(1.66±0.63)cm，体尾部(1.42±0.52)cm，P<0.05。 结论胰岛素瘤在胰腺体尾部较头颈部更好发；绝大多数单发，但可以全胰腺多发；多数小于1.5 cm，肿瘤的大小与患者年龄和肿瘤的解剖分布相关。     

Pathogenesis of carcinoma of the papilla of Vater

Most adenomas and carcinomas of the small intestine and extrahepatic bile ducts arise in the region of the papilla of Vater. In familial adenomatous polyposis (FAP) it is the main location for carcinomas after proctocolectomy. In many cases symptoms due to stenosis lead to diagnosis at an early tumor stage. In about 80%, curative intended resection is possible. Operability is the most relevant prognostic factor. Most ampullary carcinomas resp. carcinomas of the papilla of Vater develop from adenomatous or flat dysplastic precursor lesions. They can be sited in the ampulloduodenal part of the papilla of Vater, which is lined by intestinal mucosa. They also can develop in deeper parts of the ampulla, which are lined by pancreaticobiliary duct mucosa. Intestinal-type adenocarcinoma and pancreaticobiliary-type adenocarcinoma represent the main histological types of ampullary carcinoma. Furthermore, there exist unusual types and undifferentiated carcinomas. Many carcinomas of intestinal type express the immunohistochemical marker profile of intestinal mucosa (keratin 7?, keratin 20+, MUC2+). Carcinomas of pancreaticobiliary type usually show the immunohistochemical profile of pancreaticobiliary duct mucosa (keratin 7+, keratin 20?, MUC2?). Even poorly differentiated carcinomas, as well as unusual histological types, may conserve the marker profile of the mucosa they developed from. These findings underline the concept of histogenetically different carcinomas of the papilla of Vater which develop either from intestinal- or from pancreaticobiliary-type mucosa of the papilla of Vater. Molecular alterations in ampullary carcinomas are similar to those of colorectal as well as pancreatic carcinomas, although they appear at different frequencies. In future studies, molecular alterations in ampullary carcinomas should be correlated closely with the different histologic tumor types. Consequently, the histologic classification should reflect the histogenesis of ampullary tumors from the two different types of papillary mucosa.     

Demonstration of the mechanism of action of biguanides

Summary Palmitic acid oxidation in rat diaphragm homogenate is depressed by biguanide concentrations that are still incapable of inhibiting oxidative phosphorylation. Glucose oxidation is not directly effected by the same biguanide concentrations: however, the inhibitory effect of palmitic acid on glucose oxidation is partly removed by biguanides. Inhibition of fatty acid oxidation, which accounts for most of the metabolic effects caused by these drugs, can be regarded as the fundamental mechanism of action of biguanides. There is some evidence suggesting that these drugs might interact with carnitine, thus preventing long-chain fatty acids from being transported across the mitochondrial membrane to the site of oxidation. Traduzione a cura degli AA.     

Lack of correlation of degree of villous atrophy with severity of clinical presentation of coeliac disease

BACKGROUND AND AIM: Both the clinical presentation and the degree of mucosal damage in coeliac disease vary greatly. In view of conflicting information as to whether the mode of presentation correlates with the degree of villous atrophy, we reviewed a large cohort of patients with coeliac disease. PATIENTS AND METHODS: We correlated mode of presentation (classical, diarrhoea predominant or atypical/silent) with histology of duodenal biopsies and examined their trends over time. RESULTS: The cohort consisted of 499 adults, mean age 44.1 years, 68% females. The majority had silent coeliac disease (56%) and total villous atrophy (65%). There was no correlation of mode of presentation with the degree of villous atrophy (p=0.25). Sixty-eight percent of females and 58% of males had a severe villous atrophy (p=0.052). There was a significant trend over time for a greater proportion of patients presenting as atypical/silent coeliac disease and having partial villous atrophy, though the majority still had total villous atrophy. CONCLUSIONS: Among our patients the degree of villous atrophy in duodenal biopsies did not correlate with the mode of presentation, indicating that factors other than the degree of villous atrophy must account for diarrhoea in coeliac disease.     

血吸虫童虫生长发育及其机制的研究进展

血吸虫童虫是宿主免疫系统攻击的重要靶标,包括皮肤型、肺型和肝门型童虫。宿主分子对童虫生长发育具有重要作用。童虫生长发育机制包括免疫调节、信号转导、性别发育及凋亡等。肌动蛋白、组织蛋白酶、烯醇化酶和葡萄糖基转移酶等分子为血吸虫童虫生长发育的重要分子。本文对血吸虫童虫生长发育及其机制的研究进展做一综述。     

124例耐多药结核分枝杆菌基因突变特征分析

目的对临床分离的耐多药结核分枝杆菌相关基因的突变特征进行分析。方法对124例耐多药结核分枝杆菌以及50株敏感株的耐药相关基因(包括异烟肼inh A、kat G、oxyR-ahp C间隔区以及利福平rpo B)进行序列测定,分析其基因突变情况。结果异烟肼耐药inh A基因突变率为14.5%;kat G基因突变率为70.2%(87/124),主要位于315位;oxyR-ahp C间隔区突变率为15.3%;inh A、kat G两种基因同时突变率75.0%,三种基因同时突变率为89.5%。利福平rpo B基因突变的检出率高达95.2%,突变主要发生在531、526、516位点。结论我省耐多药菌异烟肼耐药相关基因最常见突变为kat G 315、inh A C-T(-15)、axyR-ahp C间隔区(-10)C-T,利福平为rpo B531、526、516。结合MDR-TB耐药相关基因的特征分析,可以建立一种快速、准确、特异的适合于我省的检测结核菌耐多药性的新方法。     

氯硝柳胺悬浮剂的毒性评价

目的评价氯硝柳胺悬浮剂的毒性，为现场大规模应用灭螺提供依据。方法按照中华人民共和国国家标准GB 15670-1995《农药登记毒理学试验方法》和鱼类毒性试验方法进行。结果经口、经皮肤的LDso雌、雄性大鼠均>5 000 mg/kg，经呼吸道的LCso雌、雄性大鼠均>5 000mg/m3，该药经口、经皮肤、经呼吸道毒性均属微毒类药物；兔眼用药后，观察期内无不良反应，对眼无刺激性；皮肤用药后对皮肤无刺激性。与氯硝柳胺原药、氯硝柳胺乙醇胺盐原药和氯硝柳胺乙醇胺盐可湿性粉剂相比，氯硝柳胺悬浮剂对鱼急性毒性最低。结论氯硝柳胺悬浮剂属微毒类药物，对鱼的毒性低于其乙醇胺盐可湿性粉剂，适合于现场应用。     

Assessment of quality of life of parents of children with juvenile chronic arthritis

The aim of the study was to assess the quality of life (QOL) and the psychological status of parents of children with juvenile chronic arthritis (JCA). The QOL, anxiety and depression of the parents of 28 children with JCA were evaluated and compared to those of the parents of 28 healthy children. Mothers of JCA children and mothers of healthy children reported similar QOL. The reported anxiety and depression levels were similar for mothers and fathers in both groups. The parents of children with pauciarticular-type JCA reported lower QOL and higher levels of anxiety and depression than the parents of children with other types, namely polyarticular and systemic JCA. These findings may be explained by the fact that the pauciarticular patients had shorter disease duration and were less frequently seen in the outpatient clinic. The QOL of mothers of children with JCA was found to be slightly impaired in the group of children with pauciarticular JCA. Future larger studies are needed to confirm these results, as the number of subjects in the three groups was rather low. Received: 26 September 2001 / Accepted: 8 February 2002     

Numerical Simulation of the Effect of Rate of Change of Glucose on Measurement Error of Continuous Glucose Monitors

Background

A 5-day in-patient study designed to assess the accuracy of the FreeStyle Navigator® Continuous Glucose Monitoring System revealed that the level of accuracy of the continuous sensor measurements was dependent on the rate of glucose change. When the absolute rate of change was less than 1 mg•dl⁻¹•min⁻¹ (75% of the time), the median absolute relative difference (ARD) was 8.5%, with 85% of all points falling within the A zone of the Clarke error grid. When the absolute rate of change was greater than 2 mg•dl⁻¹•min⁻¹ (8% of the time), the median ARD was 17.5%, with 59% of all points falling within the Clarke A zone.

Method

Numerical simulations were performed to investigate effects of the rate of change of glucose on sensor measurement error. This approach enabled physiologically relevant distributions of glucose values to be reordered to explore the effect of different glucose rate-of-change distributions on apparent sensor accuracy.

Results

The physiological lag between blood and interstitial fluid glucose levels is sufficient to account for the observed difference in sensor accuracy between periods of stable glucose and periods of rapidly changing glucose.

Conclusions

The role of physiological lag on the apparent decrease in sensor accuracy at high glucose rates of change has implications for clinical study design, regulatory review of continuous glucose sensors, and development of performance standards for this new technology. This work demonstrates the difficulty in comparing accuracy measures between different clinical studies and highlights the need for studies to include both relevant glucose distributions and relevant glucose rate-of-change distributions.     

治疗高血压药物的经济学评价

重视高血压治疗中的经济学评价,对利用我国有限的卫生资源来遏制高血压对人民群众的危害有着重要的现实意义。药物经济学对于药物治疗的成本和治疗的结果给予同样的关注。因为治疗高血压的费用,不仅涉及药物价格,还包括患者的危险水平,降压疗效和对临床终点事件的影响,以及治疗的依从性和安全性。因此药物经济学更强调整体成本和价-效比。低危病人,若非药价低廉,治疗的价-效比不够理想。而在高危的患者,价-效比越小越经济而不是药费越便宜越好。