SARS冠状病毒半胱氨酸蛋白酶3CLpro的克隆表达及分离纯化

SARS（严重急性呼吸综合征）是由一种新型的人类冠状病毒引起的。由于SARS冠状病毒3CL蛋白酶在病毒的复制翻译中起关键作用，所以成为抗SARS药物设计的关键靶标之一。该研究中，利用以pET28a为载体的高效原核表达系统，克隆表达出SARS冠状病毒3CL蛋白酶，然后由NTA-Ni^2＋亲和层析柱对表达的蛋白进行分离纯化。3CL蛋白酶在大肠杆菌表达系统中的成功表达为今后进一步筛选抗SARS病毒药物奠定基础。     

甲磺酸伊马替尼治疗国人胃肠间质瘤的临床研究

 目的 观察和评价甲磺酸伊马替尼（Imatinib mesylate）治疗国人胃肠间质瘤（GIST）的有效性和安全性。方法 2002年8月至2004年12月，在本院通过病理形态学及免疫组化确诊的GIST共52例，其中36例应用伊马替尼治疗，用法为伊马替尼400mg，口服，1／日。参照WHO实体瘤客观疗效标准观察和判定疗效，NCI-CTC2．0版抗癌药的毒性标准观察和判定毒性。结果 伊马替尼治疗的36例患者中，包括新辅助治疗和姑息治疗在内可以评价疗效的有28例，用药后获得部分缓解为14例（50％），疾病稳定10例（35．7％），疾病进展4例（14．3％）；即有效率（RR）为50％，疾病控制率（DCR）达到85．7％。36例均可进行毒性评价，除了1例因胃部GIST合并脾脏B细胞型非霍奇金淋巴瘤，姑息切除术后口服格列卫同时进行CHOP方案化疗，结果 发生了Ⅲ级骨髓抑制外，其他35例中毒副反应均为Ⅰ～Ⅱ级，而且大多数毒性可以控制或恢复正常。结论 与国外文献报道一致，甲磺酸伊马替尼治疗国人GIST安全高效，耐受性好。     

Current status and future prospects of chemotherapy for metastatic gastric cancer: a review

Although many randomized trials of chemotherapy for metastatic gastric cancer have been reported during the past two decades, no standard regimens worldwide have been established yet. Reference arms vary depending on the region and cultural differences. To date, a combination of 5-fluorouracil (5-FU) and cisplatin is most widely used. However, no confirmation of survival advantage over single-agent 5-FU in a randomized trial has been proved yet, and there remain limitations of efficacy results in older-generation regimens. Recently developed new agents such as irinotecan, taxanes (paclitaxel and docetaxel), and new oral fluorouracil (S-1 and capecitabine) provided more promising results: a response rate over 50% and median survival time (MST) over 10 months in their preliminary combination studies. These newer combination regimens are now being investigated in various randomized phase III studies, which will clarify whether the newer-generation regimens provide survival advantage over older-generation regimens. The MST of the new standard should exceed 11 months to be considered a definite improvement, and overall survival seems to be a more desirable primary end point than progression-free survival in a randomized trial. Molecular targeting agents are another concern to improve the treatment outcomes of this disease and are now under investigation in combination with conventional cytotoxic agents. Both clinical and biological research will be more important in future studies.     

Molecular weights of poly(butyl cyanoacrylate) nanoparticles determined by mass spectrometry and size exclusion chromatography

Matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS) and size exclusion chromatography (SEC) were employed to elucidate the chemical composition, mean number average molecular weight (M_n), mean weight average molecular weight (M_w), and polydispersity (PD) of poly(butyl cyanoacrylate) (PBCA) manufactured by emulsion polymerisation. Both methods gave similar results for M_n, but substantial differences were observed for M_w and PD, with MALDI producing consistently lower values which could not be improved by off-line coupling of SEC and MALDI. MALDI gave a more detailed view on the chemical composition of the cyanoacrylate and revealed the presence of two additional polymer series with different end groups besides the expected PBCA series, which showed different retention in SEC. Their formation is explained by the secession/addition of formaldehyde from/to the regular polymer via (reverse) Knoevenagel reaction. In additional experiments, the influence of different pH on PBCA-NP during polymerisation was examined by comparison of polymerisation yield and particle diameter to their chemical composition as revealed by the MALDI spectra. The most uniform nanoparticles, with the highest polymerisation yield, narrowest particle size, and mass distribution were produced at pH 1.     

NMR studies of the inclusion complex between β-cyclodextrin and paroxetine

A ¹H and ¹³C NMR study on the inclusion complex of paroxetine with β-cyclodextrin was carried out in order to define the stoichiometry of the association and its strength. Proton and carbon chemical shift measurements of paroxetine and β-cyclodextrin were performed at several molar ratios and temperatures, allowing the determination of a 1:1 stoichiometry and an association constant value of the order of 2 × 10³ for the paroxetine–β-cyclodextrin complex. Overhauser effects in the rotating frame were also measured, and the experimental interproton distance constraints have been used for molecular model building of the complex. The obtained model indicates that the benzodioxolyl moiety of paroxetine is deeply inserted in the cavity of the cylindrical structure of β-cyclodextrin, while the fluoro-phenyl ring lays above the wider rim.     

CCLG-ALL2008方案治疗不同分子生物学特征儿童急性淋巴细胞白血病的长期疗效分析

目的 探讨CCLG-ALL2008方案治疗具有不同分子生物学特征的初诊儿童急性淋巴细胞白血病（ALL）的长期疗效。方法 选取按照CCLG-ALL2008方案治疗的940例初诊ALL患儿为研究对象,针对不同分子生物学特征ALL的长期疗效进行回顾性分析。结果 940例ALL患儿中,男570例,女370例,中位年龄5（1~15）岁,中位随访时间65（3~123）个月。完全缓解（CR）率为96.7%,预期10年总体生存（OS）率为（76.5±1.5）%,无事件生存（EFS）率为（62.6±3.0）%。患儿经治疗达CR后,总复发率为21.9%,其中ETV6-RUNX1阳性患儿复发率最低,且易于晚期复发;MLL重排阳性患儿复发率最高,且易于早期复发。ETV6-RUNX1阳性患儿的预期10年OS率明显高于伴有TCF3-PBX1阳性、BCR-ABL阳性、MLL重排及无分子生物学特征患儿（P < 0.05）。ETV6-RUNX1阳性患儿的预期10年EFS率明显高于伴有BCR-ABL阳性和MLL重排患儿（P < 0.05）。结论 分子生物学特征是影响ALL患儿长期预后的指标,MLL重排、BCR-ABL融合基因阳性是预后的不良指标,ETV6-RUNX1融合基因阳性患儿长期生存率较高。     

Diagnóstico y seguimiento de los pacientes con hipotiroidismo congénito diagnosticados por cribado neonatal

The screening program of congenital hypothyroidism (CH) is probably one of the best achievements in paediatrics. Thyroid hormones are essential for brain development and brain maturation that continue through the neonatal period. Hypothyroidism that begins in the first months of life causes irreversible damage to the central nervous system, and is one of the most frequent and preventable causes of mental retardation. As children with congenital hypothyroidism are born with a normal appearance, analytical studies are required to immediately start the appropriate therapy.This article analyses the aims, diagnostic procedures, tests required, aetiology, and differential diagnosis in this disorder. Especially relevant is to perform frequent monitoring to ensure dose adjustments of L-Thyroxine therapy, avoiding infra- or supra-dosing that negatively affects neurosensory functions. Re-evaluation of the aetiology permanent vs transient hypothyroidism is always recommended after 3 years of chronological age.The relevance of this screening program should be widely discussed in paediatrics. The main objective is to avoid cerebral damage in these patients, and has been highly successful and economically beneficial.Other aspects are required to optimise patient outcomes, to perform all the controls according to the recommendations and to include, in the near future, the diagnosis of central hypothyroidism. Implementation of this program is necessary to progress in accordance with current scientific knowledge.     

The association between COX-2 gene rs5275 polymorphism and Nasopharyngeal carcinoma risk

The overexpression of cyclooxygenase-2 (COX-2) was correlated with the invasion and lymphatic metastasis and with the clinical stage of Nasopharyngeal carcinoma (NPC). The C allele of COX-2 gene rs5275 polymorphism disrupts miR-542-3p function to promote COX-2 overexpression. To examine the role of COX-2 gene rs5275 polymorphism in NPC, we determined COX-2 gene rs5275 polymorphism by using a custom-by-design 48-Plex single nucleotide polymorphism (SNP) Scan^? Kit. We found that C allele or CC genotype of rs5275 polymorphism in COX-2 gene was associated with an increased risk of NPC. In stratified analyses, COX-2 gene rs5275 polymorphism was associated with the risk of NPC among females, smokers, and drinkers. Based on these results, we concluded that COX-2 gene rs5275 variant contributes to NPC risk in a Chinese population. Larger studies with more diverse ethnic populations are needed to confirm these results.     

Use of the Ion AmpliSeq Cancer Hotspot Panel in clinical molecular pathology laboratories for analysis of solid tumours: With emphasis on validation with relevant single molecular pathology tests and the Oncomine Focus Assay

Targeted application of next-generation sequencing (NGS) technology allows detection of specific mutations that can provide treatment opportunities for cancer patients. We evaluated the applicability of the Ion AmpliSeq Cancer Hotspot Panel V2 (CHV2) using formalin-fixed, paraffin-embedded (FFPE) tissue of clinical specimens.Thirty-five FFPE tumour samples with known mutational status were collected from four different hospitals and sequenced with CHV2 using an Ion Chef System and Ion S5 XL system. Out of 35 cases, seven were sequenced with Oncomine focus Assay Panel for comparison. For the limit of detection test, we used an FFPE reference standard, a cell line that included an engineered 50% EGFR T790?M in an RKO cell line background. Coverage analysis results including number of mapped reads, on target percent, mean depth, and uniformity were not different according to hospitals. Sensitivity for mutation detection down to 3% was demonstrated. NGS results showed 100% concordance with the results from single molecular pathology tests Assay in 30 cases with 24 known positive mutations and 14 known negative mutations, and another NGS panel of the Oncomine focus in seven cases.The CHV2 NGS test for solid tumours using Ion chef system and S5 XL system in clinical molecular pathology laboratories for analysis of solid tumours could be routinely used and could replace some single molecular pathology tests after a stringent and thorough validation process.     

“Hepatocellular carcinoma: A life-threatening disease”

An estimated rise in liver cancer incidence will increase to 95374 new cases by 2020. Hepatocellular Carcinoma (HCC), the most common primary malignant tumour of the liver, is considered to be the third leading cause of all cancer-related deaths and fifth common cancer worldwide. The reported data shows that the rate of HCC incidence in male population is three to four times higher compared with the female population. In the United States, HCV-induced liver cancer is increasing very fast because of the lack of proper treatment option. There are various treatment strategies available for HCC like liver transplantation, resection, ablation, embolization and chemotherapy still the prognosis is destitute. If the patient is eligible, liver transplantation is the only therapeutic option that may give around 90% survival rate, but the scarcity of liver donor limits its broad applicability. A sudden address is necessary to develop specific drugs, personalized medicine, for HCC.