一种新的编码霍乱毒素的丝状噬菌体CTAKΦ

目的 探讨介导霍乱弧菌毒素基因水平转移的一种独特的遗传结构。方法 从O139群霍乱弧菌菌株FJ97129上清中分离出丝状噬菌体颗粒CTAKΦ，并进行电镜观察。从丝状噬菌体颗粒CTAKΦ中纯化出DNA，并进行链型分析。对pCTAK进行酶谱分析。用ctxAB、zot引物对pCTAK进行PCR扩增检测，用RS1探针对pCTAK进行Southern blot检测；用pCTAK转化DH5α和IEM101得到DX29和4329，将pCTAK克隆于pUC18载体并转入DH5α得到UD29，用GM1－ELISA检测DX29、4329、UD29的CT表达。对pCTAK的ctxAB、zot基因片段进行测序，并用DNASTAR软件和BLAST算法，在国际互联网上对测序结果进行序列分析。结果 发现和分离了一种编码霍乱毒素的质粒pCTAK，它以稳定的高拷贝数存在于1株天然的O139霍乱弧菌FJ97129中；酶谱分析发现其明显不同于CTX元件酶谱，在CTX元件中相当保守的酶切位点：BglⅡ、EcoRⅤ、PstⅠ、EcoRⅠ，在pCTAK中没有切点。ctxAB、zot引物对pCTAK的PCR扩增检测呈阳性，RS1探针杂交呈阳性；pCTAK的ctxAB、zot基因序列与已报道的序列有很高的同源性。pCTAK能直接或经载体转化DH5α和IEM101并表达CT。从FJ97129培养上清中分离出丝状噬菌体颗粒CTAKΦ，电镜观察并计算其直径大小为7nm左右。其全基因组大小为8.5kb，为单链DNA。结论 发现了一种新的编码霍乱毒素的丝状噬菌体CTAKΦ。     

Hypophosphatemic osteomalacia: a report of five cases and evaluation of bone markers

In this study, we analyzed the changes in biochemical markers of bone turnover in five patients with hypophosphatemic osteomalacia. The following bone markers were evaluated: among bone formation markers, total alkaline phosphatase (TAP), bone alkaline phosphatase (BAP), osteocalcin (bone Gla protein, BGP) and procollagen type I N propeptide (PINP); among bone resorption markers, serum C-terminal cross-linked telopeptide of type I collagen (s-CTx), urinary hydroxyproline (HYP), and N-terminal and and C-terminal cross-linked telopeptides of collagen (NTx and - and -CTx). In addition, the /-CTx ratio was evaluated. TAP and BAP were the markers with the highest increase in both frequency and magnitude. Conversely, BGP values were low in all patients. Collagen-related markers were slightly increased in nearly half of the patients. Among them, PINP showed the highest proportion of increased values. The /-CTx ratio was within normal values in all patients. In conclusion, TAP and BAP seem to be the best bone markers in the diagnostic evaluation of hypophosphatemic osteomalacia. In addition, their high values associated with low levels of BGP provide an even more reliable biochemical profile of this disorder, when associated with the classic mineral and skeletal homeostasis abnormalities.     

抗ENO1抗体联合二甲双胍通过靶向肿瘤干细胞逆转人非小细胞肺癌A549细胞对西妥昔单抗的抵抗

目的:探究抗烯醇化酶1 (enolase 1,ENO1)抗体、二甲双胍(metformin,MET)通过靶向肿瘤干细胞对西妥昔单抗(cetuximab,CTX)耐药型非小细胞肺癌A549细胞增殖、迁移、侵袭和干性的影响及其可能的作用机制.方法:10 mmol/L MET联合40μg/ml抗ENO1抗体处理CTX(35 ...     

Immunomodulatory effects of diclofenac in leukocytes through the targeting of Kv1.3 voltage-dependent potassium channels

Kv1.3 plays a crucial role in the activation and proliferation of T-lymphocytes and macrophages. While Kv1.3 is responsible for the voltage-dependent potassium current in T-cells, in macrophages this K⁺ current is generated by the association of Kv1.3 and Kv1.5. Patients with autoimmune diseases show a high number of effector memory T cells that are characterized by a high expression of Kv1.3 and Kv1.3 antagonists ameliorate autoimmune disorders in vivo. Diclofenac is a non-steroidal anti-inflammatory drug (NSAID) used in patients who suffer from painful autoimmune diseases such as rheumatoid arthritis. In this study, we show that diclofenac impairs immune response via a mechanism that involves Kv1.3. While diclofenac inhibited Kv1.3 expression in activated macrophages and T-lymphocytes, Kv1.5 remained unaffected. Diclofenac also decreased iNOS levels in Raw 264.7 cells, impairing their activation in response to lipopolysaccharide (LPS). LPS-induced macrophage migration and IL-2 production in stimulated Jurkat T-cells were also blocked by pharmacological doses of diclofenac. These effects were mimicked by Margatoxin, a specific Kv1.3 inhibitor, and Charybdotoxin, which blocks both Kv1.3 and Ca²⁺-activated K⁺ channels (K_Ca3.1). Because Kv1.3 is a very good target for autoimmune therapies, the effects of diclofenac on Kv1.3 are of high pharmacological relevance.     

中西医结合治疗特发性膜性肾病临床观察

目的 :观察中西医结合治疗特发性膜性肾病临床疗效。方法 :对 6 8例特发性膜性肾病随机分为对照组 32例 ,治疗组 36例 ;对照组单纯用强的松和环磷酰胺、潘生丁、洛丁新 ,治疗组在用上述西药同时辨证加用中药治疗 ,观察 3年～ 6年。结果 :对照组完全缓解率为 2 8.1% ,总缓解率为 6 5 .6 % ;治疗组完全缓解率为 5 5 .6 % ,总缓解率为 86 .2 %。治疗组完全缓解率和总缓解率明显优于对照组 ,两组比较P <0 .0 5 ,具有显著性差异。结论 :中西医结合是治疗膜性肾病的有效方法。     

环磷酰胺冲击治疗狼疮肾炎的临床研究

目的通过临床对比研究，探讨应用环磷-酰胺冲击治疗（IV－CTX）狼疮肾炎（LN）的最佳方案。方法将92例LN患者随机分成3组：A组，IV－CTX每2周1次，每次（8～12）mg／kg，连用2天，B组，IV－CTX每月1次，每次（0．5～1．0）g／m2；C组，IV－CTX每3个月1次，每次（0．5～1．0）g／m2。3组均同时口服波尼松。结果A组起效时间显著比B组与C组快；病情缓解率A组也显著高于B组及C组（P＜0．01）；病情活动性积分下降至25％、50％和75％时，3组CTX累积量无显著差异（P＞0．05）；3组间不良反应及其发生率无显著统计学差异（P＞0．05）。结论IV－CTX治疗LN应根据狼疮活动程度选择。在急重症LN，应用A组2周1次方案有利于及时控制狼疮活动，提高疗效，保护肾功能；在LN病情基本控制后，改用B组每月1次方案可以巩固疗效；在LN病情完全控制后，应用C组3个月1次方案可以预防复发。     

大剂量环磷酰胺冲击疗法治疗难治性天疱疮的护理2例

我科 2 0 0 1年收治了 2例病情严重、治疗困难的天疱疮患者 ,常规治疗无效 ,采用特大剂量ＣＴＸ治疗并精心护理 ,使病情得以控制 ,报告如下。1　病例介绍例 1:患者 ,男 ,6 0岁 ,干部。因全身皮肤反复发生水疱、糜烂 5ａ,于 2 0 0 1年 5月第 6次入院。患者于 1995年开始 ,反复全身出现水疱、糜烂及口腔溃疡 ,1996年 8月第 1次入院 ,皮损病理 :表皮内疱 ,棘细胞松解 ,棘细胞间ＩｇＧ、Ｃ3 呈网状沉积。诊断 :寻常型天疱疮。经激素及ＣＴＸ等治疗 ,病情缓解 ,皮损全部消退出院。 1998年底喝酒后病情复发而再次住院 ,经 3个疗程小剂量激素冲击并…     

CTX-M-14 type β-Lactamase producing Escherichia coli isolated from hospitalized patients in Tunisia

Escherichia coli (E.coli) with a CTX-M resistance phenotype was selected from hospitalized patients in a university Hospital of Tunisia between January 2010 and June 2010. PCR analysis and sequencing demonstrated that it harboured CTX-M-14 β-lactamase. Characterization of the regions surrounding the bla(CTX-M-14) showed the ISEcp1 elements located in the upstream region of the bla gene. PFGE and multilocus sequence typing revealed two different types which corresponds to sequence types ST38 complex and ST131. These results reinforce the potential for spreading of this gene among E. coli clinical strains in the coming years in Tunisia.     

鸡血藤提取物对环磷酰胺致白细胞低下大鼠的影响

目的利用环磷酰胺(CTX)致白细胞低下大鼠模型研究鸡血藤提取物的升白细胞作用,发现其升白细胞作用的有效部位。方法采用CTX(100 mg/kg)单次sc的方法制备大鼠白细胞低下模型,给药后第7、9、11天采集大鼠的血液,测定白细胞总数及分类,并进行统计分析。结果鸡血藤提取物A(50%乙醇提取,为总提取物)、B(50%乙醇提取-水洗脱)、C(50%乙醇提取-25%乙醇洗脱)组均未见明显升白细胞作用,而鸡血藤提取物D(50%乙醇提取-70%乙醇洗脱,总黄酮量大于55%)和阳性药利可君均具有明显的升白细胞作用,且鸡血藤提取物D的升白细胞作用具有量效关系,优于阳性药利可君。结论鸡血藤提取物D可能是鸡血藤升白细胞作用的有效部位,推测其升白细胞作用的有效物质是总黄酮。