聚合酶链反应检测肾移植受者尿中人巨细胞病毒的临床应用

为探讨聚合酶链反应（ＰＣＲ）技术检测肾移植受者（ＲＴＲ）尿中人巨细胞病毒（ＨＣＭＶ）ＤＮＡ在临床工作中的应用及其价值，本组采用ＨＣＭＶ早期及晚期基因组两对引物及ＨＣＭＶ特异性单抗分别建立了ＰＣＲ技术及捕获、间接酶联法（ＭａｃＥＬＩＳＡ，ＩＥＬＩＳＡ）。应用ＰＣＲ检测ＲＴＲ尿中ＨＣＭＶＤＮＡ、ＭａｃＥＬＩＳＡ及ＩＥＬＩＳＡ法检测各受血者中ＨＣＭＶ－ＩｇＭ及ＩｇＧ，并比较其结果，表明：６５例ＲＴＲ尿中ＨＣＭＶＤＮＡ阳性率为６０％，ｌｇＭ及ＩｇＧ阳性者尿中ＨＣＭＶＤＮＡ阳性率各为８１．８％及５４．５％。说明ＰＣＲ应用存在一定局限性，应与血清学等方法相结合，取长补短，才能适应临床需要，充分发挥其应用价值。     

聚合酶链反应诊断结核性腹膜炎的评价

目的评价聚合酶链反应（ＰＣＲ）方法对结核性腹膜炎的诊断价值。方法用ＰＣＲ技术检测３０例结核腹水中结核分支杆菌ＤＮＡ，并与腹水涂片抗酸染色及酶联免疫吸附试验（ＥＬＩＳＡ）检测腹水中抗ＰＰＤ抗体进行比较。结果ＰＣＲ的阳性率为６０％，特异性９４．４％；ＥＬＩＳＡ法阳性率６３．３％，特异性７２．２％；涂片镜检均为阴性。结论ＰＣＲ在诊断结核性腹膜炎具有较高的敏感性和特异性，优于ＥＬＩＳＡ法及涂片镜检，如与ＥＬＩＳＡ技术结合可进一步提高检测的敏感性和特异性。     

弓形虫P30基因DNA免疫小鼠诱导小鼠体液应答及保护性研究

目的 用重组质粒ｐｃＤＮＡ３－Ｐ３０免疫ＢＡＬＢ／ｃ小鼠,观察其ＤＮＡ免疫所诱导的小鼠体液免疫反应和保护性作用。方法：大量制备闰ＤＮＡ,肌肉注射免疫小鼠。ＥＬＩＳＡ测定ＩｇＧ抗体滴定,免疫鼠血液组织Ｐ３０基因ＰＣＲ扩增,弓形虫攻击感染。结果 用ＥＬＩＡＳ法检测的抗体ＩｇＧ滴度１：２５６０,免疫后３周及６个月从免疫鼠血液组织中检测到Ｐ３０基因;攻击感染实验组的存活时间较对照时间延长（Ｐ〈０．０５）,     

孕妇巨细胞病毒感染对子代影响的研究

选用抗μ链捕获法或间接ＥＬＩＳＡ法测定了８１４９例健康初孕妇（对照组）和有先天性心脏病（ＣＨＤ）家族史孕妇（３１２例）的血清ＣＭＶ－ＩｇＭ抗体，并用ＰＣＲ法检测心肌组织ＤＮＡ。结果８１４９例对照组孕妇ＣＭＶ－ＩｇＭ阳性１０４０例（１２．７６％），有ＣＨＤ家族史组３１２例孕妇ＣＭＶ－ＩｇＭ阳性７３例（２３．３９％），其中１例被干预ＣＨＤ胎儿心肌组织ＣＭＶ－ＤＮＡ阳性。提示孕妇血清ＣＭＶ监测对ＣＨＤ家族史者有重要意义。     

聚合酶链反应检测肺炎支原体的临床价值

应用聚合酶链反应（ＰＣＲ）技术检测１１４例肺炎患者咽拭子中的肺炎支原体（ＭＰ），阳性１６例，阳性率为１４．０４％；同时进行酶联免疫吸附试验（ＥＬＩＳＡ）检测。证实ＰＣＲ法比ＥＬＩＳＡ法敏感、特异。     

应用PCR,BA—ELISA早期诊断结核性胸膜炎的临床意义

应用ＰＣＲ检测６４例胸水中结核菌基因组ＤＮＡ，并与ＢＡ－ＥＬＩＳＡ检测胸水中抗ＰＰＤ抗体及细菌学检查相比较。结果：三种方法的敏感性和特异性分别为５６．０％，１００％；６４．０％，８６．７％，１１．０％和１００％。若以ＰＣＲ，ＢＡ－ＥＬＩＳＡ任一阳性作为判断标准则敏感明显提高至８９．０％。认为，ＰＣＲ与ＢＡ－ＥＬＩＳＡ诊断结核性胸膜炎具有较高的敏感性，但前者更具早期和特异性并对疾病的疗效和预后有判断     

应用PCR、BA－ELISA早期诊断结核性胸膜炎的临床意义

应用ＰＣＲ检测６４例胸水中结核菌基因组ＤＮＡ，并与ＢＡ－ＥＬＩＳＡ检测胸水中抗ＰＰＤ抗体及细菌学检查相比较。结果：三种方法的敏感性和特异性分别为５６．０％、１００％；６４．０％、８６．７％、１１．０％和１００％。若以ＰＣＲ、ＢＡ－ＥＬＩＳＡ任一阳性作为判断标准则敏感性明显提高至８９．０％。认为，ＰＣＲ与ＢＡ－ＥＬＩＳＡ诊断结核性胸膜炎具有较高的敏感性，但前者更具早期和特异性并对疾病的疗效和预后有判断意义。     

PCR技术诊断巨细胞病毒宫内感染的临床价值

采用聚合酶链反应（ＰＣＲ）技术及酶联免疫吸附试验（ＥＬＩＳＡ）法对６６例血清抗居细胞病毒抗体ＩｇＭ（ＣＭＶ－ＩｇＭ）阳性孕妇的新生儿脐血进行了巨细胞病毒脱氧核糖核酸（ＣＭＶ－ＤＮＡ）与ＣＭＶ－ＩｇＭ检测。结果显示，其ＣＭＶ－ＤＮＡ阳性率明显高于ＣＭＶ－ＩｇＭ阳性率，差异非常显著。表明用ＰＣＲ技术检测胎儿脐血ＣＭＶ－ＤＮＡ有助于巨细胞病毒（ＣＭＶ）宫内感染的早期诊断，且ＣＭＶ宫内感染影响胎儿发生发育     

多项免疫吸附凝集试验检测孕妇弓形虫感染的实验研究

用弓形虫ＩｇＧ／ＩｇＡ／ＩｇＭ免疫吸附凝集试验检测５０７份孕妇血清，并与ＤＡ、ＳＦＴ、ＩＳＡＧＡ－ＩｇＭＥＩＡ－ＩｇＭ多项方法检测结果比较分析。多项检测结果阴性的２０１份血清中，该法检测出ＩｇＧ阳性２１份，ＩｇＡ、ＩｇＭ均为阴性，较ＤＡ敏感；多项检测结果为非活动性弓形虫感染的２０３份血清中，该法检出ＩｇＧ阳性１９７份、ＩｇＡ阳性１１份、ＩｇＭ阳性４份，阳性率分别为９７．０５％、５．４１％、１．９７％两者结果基本相符；多项检测结果为活动性感染的１０３份血清中，该法检出ＩｇＭ阳性８７份，ＩｇＡ阳性６１份，ＩｇＧ阳性１０３份，其中ＩｇＭ阳性与单项的ＩＳＡＧＡ－ＩｇＭ阳性结果相符率较高为８３．０２％。弓形虫ＩｇＧ／ＩｇＡ／ＩｇＭ免疫吸附凝集试验盒可作弓形虫感染的定性、定量检测，对判定孕妇的弓形虫感染时间是较好的方法。     

聚合酶链反应检测弓形虫的实验研究

目的建立敏感、特异、稳定的聚合酶链反应（ＰＣＲ）法,以检测弓形虫感染。方法建立ＰＣＲ法,确定最佳扩增条件;用ＰＣＲ法检测实验感染家兔血液中ＤＮＡ的动态变化,并与ＥＬＩＳＡ法检测ＣＡｇ进行比较。结果兔感染弓形虫后第２ｄ,ＰＣＲ开始出现阳性,阳性率为７６．９％（１０／１３）,总检出率达１００％。与ＥＬＩＳＡ法查ＣＡｇ进行平行检测,前者出现阳性时间早,且阳性率明显高于后者。该法敏感性高,可检测到１０ｆｇＤＮＡ含量;特异性强,对其它９种寄生虫和微生物ＤＮＡ均无交叉现象;稳定性好,对阳、阴性同一样品重复５次,结果一致。结论提示本法可用于临床诊断,在一定范围内替代病原学和免疫学检测。     

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。     

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。     

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。     

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.     

Study of constancy of hydrogen-consuming flora of human colon

The constancy of the hydrogen consuming flora of the human colon was studied in 15 healthy subjects via two measurements obtained 18 to 36 months apart. Hydrogen disappearance rate and the major products of H₂-consuming bacteria, methane and sulfide, were measured during incubation of fecal homogenates with excess hydrogen and sulfate. In 11/15, the hydrogen consumption rate and the predominant hydrogen-consuming pathway (methanogenesis, sulfate reduction, or neither) remained constant. However, major shifts in these pathways were observed in four subjects, with two losing and two gaining the ability to produce methane. Methanogenesis was associated with the highest hydrogen consumption rate. This study demonstrates that clinically unrecognizable, major alterations of the colonic flora occur in healthy subjects. Understanding of the factors responsible for these alterations might allow for therapeutic manipulation of the colonic flora.Supported in part by the Department of Veterans Affairs and NIDDKD RO1 DK 13309-25.