生物安全柜的控制

生物安全柜是重要的一级屏障设备，适当的控制方式对生物安全柜的防护能力是很重要的。文中主要介绍了生物安全柜的控制要点和几种控制方式及其特点。     

甲状腺乳头状癌基因突变与其侵袭性相关性研究

  目的  收集就诊于昆明医科大学第三附属医院的甲状腺乳头状癌（papillary thyroid carcinoma,PTC）患者的临床、病理及基因资料并整理分析,探究PTC基因突变与其侵袭性的相关性,并探讨BRAF V600E单基因突变与中央区淋巴结隐匿性转移的相关性,以期细化PTC的精准治疗。  方法  选择2018年4月至2022年4月就诊于昆明医科大学第三附属医院符合入组标准且同意入组的PTC患者,收集入组患者临床病理及基因资料并整理分析:（1）各基因突变作观察组、野生型作对照,对比各基因型突变与野生型的差异;（2）多基因突变作观察组,单基因突变作对照,对比多基因突变与单基因突变的差异; （3）cN0患者中:BRAF V600E单基因突变作观察组,无基因突变作对照,分析BRAF V600E突变与中央区淋巴结隐匿性转移的相关性。  结果  （1）BRAFV600E突变:中央区淋巴结转移（Central Lymph Node Metastasis CLNM）、N1占比大,转移淋巴结数目多,TPO-Ab降低（P < 0.05）;TERT突变:高龄、男性、侧颈淋巴结转移占比大,侧颈淋巴结转移数目多,T分期晚（P < 0.05）;RET突变:侧颈淋巴结转移占比大、侧颈转移淋巴结数目多,T分期晚（P < 0.05）,NTRK3与ETV6均融合突变,结果一致:突变组侵犯甲状腺包膜占比大（P < 0.05）;RAS突变:Tg-Ab水平低（P < 0.05）;TP53,CCDC6:突变组与野生型组未见明显差异（P≥0.05）;（2）多基因突变组侵犯包膜、侧颈淋巴结转移占比大、侧颈转移淋巴结数量多,M分期晚（P < 0.05）;3.cN0患者中:BRAF V600E单基因突变组CLNM占比大,中央区转移淋巴结数目较多（P < 0.05）。  结论  （1）PTC中,BRAF V600E突变与中央区淋巴结转移相关;（2）多基因突变较单基因突变表现出更强的侧颈淋巴结及远处转移能力;（3）cN0 PTC患者中,BRAF V600E单基因突变与隐匿性中央区淋巴结转移有关。     

318例原发性胃癌转移及预后影响因素的相关分析

  目的  探索并分析原发性胃癌（gastric cancer,GC）患者转移及预后的影响因素,寻找可早期预测转移的临床指标。  方法  回顾性研究云南省肿瘤医院2018年1月至2021年12月期间收治的经病理确诊为原发性GC的318例患者的临床病理资料,根据是否发生转移分为转移组（213例）和未转移组（105例）。分析各临床病理因素与GC转移及预后的关系,并探索各影响因素对转移的诊断效能。  结果  （1）生存分析提示转移组中位总生存期（overall survival,OS）及平均无进展生存期（progression-free survival,PFS）分别为27个月和22.16个月,未转移组中位OS及平均PFS分别为45个月和41.98个月,2组间OS及PFS,差异均有统计学意义（P < 0.0001）;（2）多因素分析提示浸润深度 > 黏膜层、未接受化疗及糖类抗原-125（carbohydrate antigen-125,CA-125）≥14.4 KU/L是GC患者发生转移的独立危险因素（P < 0.05）;（3）绘制受试者工作特征曲线（receiver operating characteristic curve,ROC曲线）提示与单指标或双指标联合相比,浸润深度、前白蛋白和CA-125三指标联合后对预测GC患者是否发生转移具有更精确的敏感性及特异性;（4）Log-rank检验及COX多因素分析提示化疗总周期<8周期及CA-125≥14.4 KU/L是转移患者OS的独立危险因素,原发灶未接受手术治疗、初诊合并转移、病程中合并远处转移及CA-125≥14.4 KU/L是转移患者PFS的独立危险因素,差异均有统计学意义（P < 0.05）。  结论  GC患者一旦发生转移则预后极差,临床中应重视转移的早期发现及诊治,多指标联合对于诊断GC转移具有较精确的敏感性及特异性,提示对GC患者应进行积极评估,制定个体化治疗方案,以期尽早诊治转移灶,为改善患者预后,延长生存期,提高生活质量提供一定的理论基础。     

The interaction of the SARS coronavirus non-structural protein 10 with the cellular oxido-reductase system causes an extensive cytopathic effect.

The pathological mechanism of SARS-CoV infection was investigated. The gene for the SARS-CoV non-structural protein 10, which is located in the open reading frame of pp1a/pp1ab gene, was synthesized and used to screen for the specific cellular gene coding for the protein interacting with this nsp10 protein in a human embryo lung cDNA library using a yeast trap method. The results indicated that apart from the two subunits of cellular RNA polymerase complex, BTF3 and ATF5, this nsp10 protein was also able to interact specifically with the NADH 4L subunit and cytochrome oxidase II. Further study revealed that the activity of the NADH-cytochrome was altered and the inner mitochondrial membrane was depolarized in the transfected human embryo lung fibroblast by the nsp10 protein gene. The cytopathic effect of the Coronavirus 229E strain appeared more extensive in these cells than in the control cells.     

Impact of maternal and pre-existing antibodies on immunogenicity of inactivated rotavirus vaccines

Rotavirus (RV) is a major pathogen causing severe diarrhea in infants and children aged less than 5 years. Vaccination is an economically feasible and effective strategy to prevent rotavirus infections. However, immune efficacy of live vaccines could be interfered by maternal antibodies and pre-existing antibodies of children. To develop an inactivated rotavirus vaccine (IRV), we had previously isolated a wild-type human rotavirus strain ZTR-68-A (G1P[8]) from the fecal samples of infants having severe diarrhea in a region endemic for the presence of this pathogen. In our present study, we assessed whether the presence of maternal and pre-existing antibodies in newborn BALB/c mice affected the immunogenicity of IRV administered to these animals. Our results indicate that maternal antibodies, generated from either vaccine immunization or rotavirus infection, showed partial influence with the immune responses generated by two doses of IRV vaccination. Increasing the number of immunizations can significantly improve the titer of serum neutralizing antibody and a seroconversion rate of up to 100%. In newborn mice, single-virus infection did not elicit detectable levels of serum neutralizing antibodies. After an IRV vaccination, the immune responses of these mice remained unaffected, with no significant differences in titers compared with those of control-group mice. In summary, choosing a suitable immunization dose and dosing frequency is essential for the immune effectiveness of IRV. The results of this study will provide animal experimental support for the IRV clinical research in future.     

Pretreatment with 3-methyladenine ameliorated Pseudomonas aeruginosa-induced acute pneumonia by inhibiting cell death of neutrophils in a mouse infection model

Pseudomonas aeruginosa is one of the leading causes of nosocomial infections worldwide. Clinical isolates that are resistant to multiple antimicrobials make it intractable. The interactions between P. aeruginosa and host cell death have multiple effects on bacterial clearance and inflammation; however, the potential intervention effects remain to be defined. Herein, we demonstrated that intravenous administration of 3-methyladenine before, but not after, P. aeruginosa infection enhanced autophagy-independent survival, which was accompanied by a decrease in the bacterial load, alleviation of pathology and reduction in inflammatory cytokines, in an acute pneumonia mouse model. Interestingly, these beneficial effects were not dependent on neutrophil recruitment or phagocytosis, but on the enhanced killing capacity induced by inhibiting the cell death of 3-MA pretreated neutrophils. These findings demonstrate a novel protective role of 3-MA pretreatment in P. aeruginosa-induced acute pneumonia.     

深低温冷冻大段同种异体骨移植修复四肢骨肿瘤切除后大段骨缺损

目的：分析大段同种异体骨移植在治疗四肢骨肿瘤切除后骨缺损中的疗效。方法：对30例四肢骨肿瘤．按Enneking分期原则确定切除范围，进行瘤段广泛切除，大段异体骨移植修复骨缺损，平均移植骨段13．6cm，并辅以坚强内固定，对高恶性骨肿瘤进行术前、后化疗。结果：随访4～53个月，治疗满意率为85％，主要并发症是感染、局部复发、肺转移死亡，其发生率为20％，保肢率为80％。结论：大段同种异体骨移植，在治疗四肢骨肿瘤切除后骨缺损方面是一种有效可行的方法。     

高温抑制舌癌细胞侵袭及转移机理的实验研究

目的探讨高温治癌与基质金属蛋白酶系统之间的关系,以期阐明高温抑制肿瘤细胞侵袭及转移的机理.方法对舌癌细胞Tca-8113进行体外43℃加温,通过免疫组化、流式细胞术及明胶酶谱活性分析等方法进行分析,以研究加温后该舌癌细胞MMP-9、MMP-2表达量及其活性的改变.结果加温与未加温细胞MMP-9、MMP-2均有表达,但加温后的细胞MMP-9和MMP-2蛋白表达量减少,并且加温后的细胞MMP-9和MMP-2的活性较未加温细胞明显下降.结论高温能通过调节基质金属蛋白酶(MMP-9、MMP-2)的表达量及其活性来抑制肿瘤细胞的侵袭、转移能力.     

恶性上腔静脉综合征的坐位急诊放射治疗

目的回顾性分析不能常规卧位放疗的恶性上腔静脉综合征坐位急诊放疗的疗效。方法12例患者先坐位急诊放疗6MV—X DF300CGY／次／天；总剂量DT3000CGY／10次／2周。待症状明显缓解已基本能平卧后再改用常规卧位放疗补量DT3000—4000CGY／15—20次／3—4周。结果2周内总缓解率92％(11／12)，整个放疗计划完成后总有效率100％(12／12)。结论病情较重、病期较晚无法耐受化疗和常规卧位放疗的患者先坐位急诊放疗待症状部分缓解后再改常规卧位放疗能迅速改善症状，提高生活质量，为下一步治疗创造有利条件。     

脐带血单个核细胞体外诱导成CIK细胞及其抗肿瘤效应的实验研究

目的:探索建立CIK细胞的原代培养方 法并阐明其生物学特性,为该细胞在肿瘤生物过 继免疫治疗中的运用奠定基础。方法:通过加入 4种细胞因子(IFN γ、IL 2、IL 1及OKT3)将脐带 血单个核细胞(CBMNCs)诱导成CIK细胞;利用 流式细胞仪测定细胞表型、细胞增殖情况及细胞 周期的分布;利用改良的MTT法测定效应细胞 的体外细胞毒活性。结果:四种细胞因子的联合 运用,可诱导出大量的CIK细胞,其相对百分率 增长了163倍(从0.14%到22%),增殖高峰位 于第10～12天,针对K562、Hela和YTMLC肿瘤 细胞株,CIK细胞在效靶比为64∶1时的杀伤活 性分别为78.57±3.48、77.7±1.90和81.2± 1.51,在32∶1时分别为62.00±2.31、60.83± 2.80和64.07±2.87,在16∶1时分别为51.43± 2.51、52.87±2.91和54.10±3.11,均显著高于 LAK细胞及CBMNCs,P=0.001;而在效靶比为 8∶1(25.10±2.66、21.4±1.55和28.77±3.56) 及4∶1(17.10±2.10、11.93±1.86和9.49±2.92) 时与后两者差异无统计学意义,P=0.083。结 论:1)脐带血可作为诱导CIK细胞的重要来源 2)联合运用细胞因子能诱导出大量的CIK细胞 其增殖高峰位于培养的第12天,适宜在此期进 行临床运用;3)CIK细胞较LAK细