PCR-SSP基因分型技术检出HLAB新等位基因B~*5516一例

目的　识别确认中国人群的HLA新等位基因。方法　使用PCR SSP以及以测序为基础的分型技术 ,分析HLA新等位基因和B 5 5 0 2基因顺序的差异及血清学方法分析特异性。结果　在四川成都骨髓供者中检测出一例新的HLA B等位基因。该基因和B 5 5 0 2基因顺序的差异 ,只是在外显子 2区域中 97C >A一个碱基取代 ,导致相应的密码子 33由酪氨酸变为组氨酸。血清学分析表明B 5 5 16与B2 2特异性相关。并由此建立起PCR SSP方法鉴定B 5 5 16基因。结论　四川成都骨髓库中检出的HLA B等位基因是HLA新等位基因 ,2 0 0 3年 8月已被世界卫生组织 (WHO)命名为HLA B 5 5 16。在大约 14 0 0例随机骨髓供者中 ,未发现其他带有B 5 5 16基因的个体。     

Hyperpolarization-activated Cyclic nucleotide-gated Channel and Cardiac Biological Pacemaker:Part Ⅱ

Abstract Hyperpolarization-activated cyclic-nucleotide-gated ( HCN) channels in the heart modulate cardiac automaticity via the hyperpolarization-activated cation current (named If, Ih, or Iq). Recent studies have unveiled the molecular identity of HCN (HCN 1-4) channels. HCN isoforms are unevenly expressed in the heart, even in the sinoatrial node. Features of HCN currents have been characterized in cardiac and other types of cells or in cell lines transfected with the HCN isoforms. The factors modulating Ih and the physiological significance of HCN channels in the heart have been extensively investigated in recent years. The hypothesis for transplanting and/or creating biological pacemakers to replace diseased sinoatrial and/or atrioventricular nodes has been postulated and tested in animal models. Local overexpression of HCN2 channels in the left atrium or in the left conductive bundle branch of the left ventricle via gene delivery induced significant Ih and escape rhythms during vagal stimulation in canines. In addition, implantation of human mesenchymal stem cells with overexpression of HCN2 channels to the canine left ventricular wall was associated with formation of spontaneous escape rhythms of left-sided origin during vagal-stimulation-induced sinus arrest. This preliminary data suggest that the use of HCN channels may hold great promise in the development of biological pacemakers.     

Granulocyte Colony Stimulating Factor and MT1-MMP Involved in Development of Atherosclerosis in Apolipoprotein E-Deficient Mice

Objectives Genetic deficiency of macrophage colony stimulating factor （M-CSF） in atherosclerosis-prone （apoE-/-） mice markedly reduces formation of atheroma. But Little is known about the potential effects of other colony stimulating factors（ CSF）, such as granulocyte CSF（ G-CSF）, on atherosclerosis. This study tested the hypothesis that G-CSF would be involved in development of atherosclerotic plaque. Methods apoE-/- mice fed with a Western-style diet （0. 15% cholesterol） were injected subcutaneously with recombinant human G-CSF（ 10 rag/day） dally for 9 weeks then sacrificed. The matrix metalloproteinase （MMP） 2 and MMP9 in serum of mice were measured by Gelatin Zymography analysis and c-kit and membrane typel-MMP （MT1-MMP） antigens were detected using fluorescence activated cell sorting （FACS）. Meanwhile, complete blood counts （CBC） and serum cholesterol, relative fractions of VLDL, LDL, and HDL were evaluated by spectrophotometric techniques and high performance liquid chromatography （HPLC） respectively. Atherosclerotic Lesions of the aorta were also analyzed by histological methods. Results G-CSF treatment resulted in increased proportions of circulating monocytes （ 6.9 ± 2. 2 % vs. 3.8 ± 0. 3 % ; P 〈 0. 05 ）, and decreased serum levels of total cholesterol（ 1225 ± 594 vs. 1991 ± 1009; P 〈 0. 005 ） compared to control mice. A greater proportion of bone marrow cells from G-CSF treated mice expressed MT1-MMP （ 14. 5 ± 5.5% vs. 6. 2 ± 5.0%, P 〈 0. 05 ） compared to bone marrow cells from vehicle treated mice. G-CSF treatment was also associated with smaller atheromatous plaque, and decreased oil red O staining. Conclusions G-CSF lowers serum cholesterol, increases circulating monocytes, increases bone marrow cell expression of MT1-MMP, inhibits plaque development, and decreases lipid and macrophage infiltration into developing plaque.     

Risk factors and comorbidities in primary biliary cirrhosis: a controlled interview-based study of 1032 patients

Primary biliary cirrhosis (PBC) is an autoimmune disease of unknown etiology, often associated with other autoimmune conditions. Controlled studies have so far provided conflicting data on risk factors and comorbidity rates in PBC. We enrolled patients with PBC (n = 1032) from 23 tertiary referral centers for liver diseases in the United States and random-digit-dialed controls (n = 1041) matched for sex, age, race, and geographical location. Patients and controls were administered a modified version of the US National Health and Nutrition Examination Study (NHANES III) questionnaire by trained personnel to evaluate associations between PBC and social, demographic, personal and family medical histories, lifestyle, and reproductive factors and the rates of comorbidity in affected individuals. Data indicate that having a first-degree relative with PBC (adjusted odds ratio [AOR] 10.736; 95% confidence interval 4.227-27.268), history of urinary tract infections (AOR 1.511, 95% CI 1.192-1.915), past smoking (AOR 1.569, 95% CI 1.292-1.905), or use of hormone replacement therapies (AOR 1.548, 95% CI 1.273-1.882) were significantly associated with increased risk of PBC. The frequent use of nail polish slightly increased the risk of having PBC. Other autoimmune diseases were found in 32% of cases and 13% of controls (P<0.0001). In conclusion, environmental factors, possibly including infectious agents through urinary tract infections or chemicals contained in cigarette smoke, may induce PBC in genetically susceptible individuals. Exogenous estrogens may also contribute to explain the female predominance of the disease.     

次氯酸钠对隐孢子虫卵囊在体外脱囊的影响

隐孢子虫卵囊的无菌化及体外脱囊是进行虫体体外细胞培养不可缺少的环节，本试验通过使用不同浓度次氯酸钠提前处理卵囊，在不同脱囊液的条件下比较各处理组的脱囊率。试验结果表明，未经任何处理的卵囊，在37C的温度下可发生脱囊，但脱囊率只有15．5％（脱囊时间90min）；经100％bleach（含5．25％次氨酸钠）作用30min，对卵囊的活性有很大的影响，在T＋TDC（Trypsin＋taurodeoxycholicacid）脱囊液中，其脱囊率只有17％（约正常卵囊的20％）；5％～20％的bleach对卵囊的活性没有明显的影响，反而有很强的刺激脱囊作用，卵囊经这一浓度范围的bleach处理后．即使没有酶的存在，其脱囊率也可达到80％以上。单独使用酶或bleach均有很好的脱囊作用，但两者合并使用，并不能提高新鲜卵囊的脱囊率。对用于细胞培养的卵囊，推荐用10％～20％bleach处理30min，这一处理，既可得到无菌化的卵囊，又有高的脱囊率。     

NIDDM病人胰岛素进入单核细胞能力的改变

本研究测定了NIDDM病人(n=6)和正常人(n=7)循环血中单核细胞(Mo)结合~(125)I-胰岛素总能力、胰岛素-受体复合物进入能力、胰岛素降解能力和趋溶酶体制剂氯喹对受体后过程的影响。结果显示NIDDM时Mo对胰岛素的结合能力、进入能力和降解能力均明显低于正常人。这些改变可能与NIDDM胰岛素抵抗性有关。     

PRIMARY CUTANEOUS NOCARDIA INFECTION DUE TO NOCARDIA ASTEROIDES

A 34-year-old white man was admitted to the hospital for treatment of cyclosporine toxicity. He was referred to the dermatology service for the evaluation of two lesions that had been present for 4 weeks on the dorsum of his left hand. The patient stated that he had cut his hand on a fence and later cleaned his aquarium two weeks before the skin lesions appeared. The past medical history was significant for a cadaveric renal transplant in 1984 for renal agenesis of the left kidney and obstructive nephropathy of the right kid-ney. His oral medications included methylprednisolone, dil-tiazem, ranitidine, cyclosporine, and ketoconazole. The recent addition of ketoconazole for oral thrush was felt to have caused the cyclosporine toxicity. The patient was cushingoid in appearance with promi-nent generalized hypertrichosis. The vital signs and the chest examination were normal. Skin examination revealed a 6-mm tense vesicle with surrounding erythema with an adjacent 5-mm firm linear papule (Fig. 1). There was no lymphadenopathy. Both lesions were biopsied, hemisec-tioned, and sent for routine light microscopy and for fungal, atypical mycobacterial, and bacterial cultures. Laboratory studies were significant for a blood urea ni-trogen of 56 mg/dL, creatinine of 2.6 mg/dL, WBC of 13,100/mm¹³, and a cyclosporine level of 2333 ng/mL (thera-peutic level 100–300 ng/mL). X-ray of the left hand showed no abnormality. Chest x-ray and computerized axial tomog-raphy scan revealed a widened superior mediastinum sec-ondary to mediastinal lipomatosis. Tissue cultures grew Nocardia asteroides. Bacterial and mycobacterial cultures, including atypical mycobacteria, were negative. The biopsy showed a mixed cellular infiltrate with scat-tered multinucleated giant cells and focal microabscesses. Brown and Brenn stain showed gram-positive filamentous organisms. A modified Fite stain (Fig. 2) demonstrated acid-fast filamentous organisms. The patient was diagnosed as having primary cutaneous nocardiosis with no evidence of dissemination. He was treated with trimethoprim sulfamethoxazole (TMP/SMX), one tablet orally four times daily, with resolution of the skin lesions within 3 weeks, but the patient developed neurolog-ic toxicity and elevation of his serum creatinine with this therapy. He was unable to tolerate a lower dose of TMP/SMX and the medication was changed to sulfisoxazole 500 mg orally four times daily, with plans to continue the treatment for 12 months. There has been no recurrence after almost 12 months of therapy.