土壤环境基因组技术及其在新药发现中的应用

土壤是微生物最重要的生境，土壤微生物具有极大的多样性。然而传统培养技术只能得到约1％的微生物纯培养，其余都是未被纯培养微生物。利用土壤环境基因组技术，可以把未被纯培养微生物的基因克隆到载体中并在宿主中进行表达，获得比已培养微生物丰富的代谢产物多样性，这为我们发现新颖结构先导化合物以及新药开辟了新的道路。本文介绍应用环境基因组技术构建土壤DNA文库的思路和方法。     

Cytotoxic lipid esters from Peucedanum ledebourielloides

Two cancer cell growth inhibitory esters, 1,2-dipalmitoyl-3-glucosyl glycerol (1) and 1,6-dihydroxy-hexane-bis-palmitoyl ester (2), together with arachidic acid-2-hydroxy-glycerol ester, daucosterol, and oleanolic acid, were isolated from the roots of Peucedanum ledebourielloides (Apiaceae family). The structures were determined by spectroscopic analyses. The esters 1 and 2 displayed significant activity against the SGC-7901, HT-29, and HL-60 cancer cell lines.     

免水分离超声乳化白内障摘除术临床观察

目的 :探讨免水分离术在超声乳化白内障摘除术中临床应用的有效性。方法 :随机选择白内障患者 2 0 0只眼 ,分两组。一组 (对照组 ) 95例 10 0只眼进行常规水分离术 ,二组 (观察组 ) 94例 10 0只眼 ,行免水分离术 ,所有患者均由同一医师进行超声乳化白内障摘除术联合人工晶体植入。临床观察超声乳化能量和时间 ,术后第一天、第三天、一周时视力 ,术中、术后并发症。结果 :( 1)平均超声能量Ⅰ、Ⅱ、Ⅲ、Ⅳ级核 ,一组分别是 :( 10 5 6± 4 16) %、 ( 17 88± 2 2 6) %、 ( 2 9 2 2± 3 5 4) %、 ( 3 7 88± 2 67) % ;二组分别是 ( 11 3 0± 1 89) %、 ( 18 76± 1 61) %、 ( 2 7 99± 3 5 8) %、 ( 3 9 2 2± 3 3 3 ) % ;两组平均超声能量差异无显著意义 (t =-0 5 12、 -1 496、 1 70 6、 -1 3 60 ,P =0 615、 0 14 2、 0 0 91、 0 182 ) ;平均超声时间一组分别为 ( 7 5 6±3 2 1)s、 ( 2 6 68± 5 15 )s、 ( 71 72± 17 2 2 )s、 ( 13 7 71± 3 8 60 )s ,二组分别为 ( 9 10± 1 2 0 )s、 ( 2 4 71± 4 0 9)s、 ( 76 17± 18 74)s、 ( 12 2 0 9± 3 5 14 )s ,两组平均超声时间差异无显著意义 (t =-1 42、 -1 415、 -1 3 69、 1 3 3 3 ,P =0 173、0 164、 0 174、 0 190 )。 ( 2 )术后一     

我国各地区医药制造业竞争力的比较分析

文章阐述了我国医药制造业的基本现状,对31个省市区(港、澳、台地区除外)的医药制造业竞争力,主要从市场占有率和市场竞争优势系数两个方面,进行了比较分析。市场占有率的优势主要集中在东部发达地区,但是市场竞争优势系数的优势分布较为均匀,各地区均有其自身的优势。     

Rural-urban differentials of premature mortality burden in south-west China

Background  

Yunnan province is located in south western China and is one of the poorest provinces of the country. This study examines the premature mortality burden from common causes of deaths among an urban region, suburban region and rural region of Kunming, the capital of Yunnan.     

Poststroke DNA immunization against neurite growth inhibitors is beneficial to the recovery from local cerebral ischemia in rats

BACKGROUND： Inhibitory signals, i.e. neurite growth inhibitors （NGIs）, presenting on central nervous system （CNS） myelin have been shown to play a crucial role in inhibiting lesioned axonal sprouting and leading to less functional recovery. Vaccines targeting NGIs may provide multifactorial protection against brain insults by overcoming the inhibitory effects of these NGIs and boosting the body＇s immune repair mechanisms. OBJECTIVE： To evaluate the effect of poststroke DNA immunization against NGIs on the rehabilitation for sensorimotor function of rat models of local cerebral ischemia. DESIGN： Completely randomized grouping design, and controlled experiment. SETTING： Brain Injury Research Laboratory, Department of Neurosurgery, National Neuroscience Institute, Singapore. MATERIALS： Sixty adult male Sprague-Dawley rats ranging in age from 45 to 120 days and in weight from 180 to 250 grams were provided by Animal Center of Department of Anatomy, Faculty of Medicine, National University of Singapore. pcDNA3.1（＋）-neurite growth inhibitors （pcDNA-NGIs） a gift was provided by Dr. Xiao from Department of Clinical Research, Singapore General Hospital, Singapore. METHODS： The experiment was carried out at Brain Injury Research Laboratory, Department of Neurosurgery, National Neuroscience Institute, Singapore from August 2003 to April 2005. （1）The involved rats were randomized into 3 groups： pcDNA-NGIs group （group A）, pcDNA3.1 （＋） group （group B） and model group （group C）, with 20 rats in each group. Left focal cerebral ischemia （FCI） was permanently induced through middle cerebral artery occlusion （MCAO） with the assistance of an operating microscope. Successful MCAO was determined by a 20% decrease to baseline in the ipsilateral cerebral blood flow. 100 μg of pcDNA-NGIs eluted in phosphate-buffered saline （PBS） was intramuscularly injected into the tibial muscle once a week after MCAO for 6 weeks in group A. As control, pcDNA3.1 （＋） was also administrated in the same way in group B and nothing was administrated in group C. （2） The modified neurological severity score （mNSS）, a composite of motor, sensory, reflex and balance tests, was used to test the sensorimotor deficit. The mNSS was graded on a scale of 0 - 18, i.e. normal score was 0, maximal deficit score was 18, and 1 point was warded for the inability to perform the tasks or the lack of a tested reflex. （3） The newly generated axons of corticorubral projection were traced by stereotaxic guided injection of 100 g/L biotinylated dextran amine. Rats were sacrificed two weeks after tracing, and cryostat coronal sections of midbrains （30μm） were reacted to BDA according to the manufacturer＇s instruction by the free-floating method. Images were captured on a DM RXA2 LEICA Microscope with a Spot Digital Camera system （Germany）, and the numbers of labeled axons on the denervated side in four standard coronal sections including the red nucleus were manually quantified. MAIN OUTCOME MEASURES： （1） The number of newly generated axons of corticorubral projection. （2）The improvement in sensorimotor deficit. RESULTS： All the involved 60 rats entered the stage of final analysis. （1） The number of newly generated axons of corticorubral projection of rats： Only ipsilateral axons of CRP were noted with little evidence of fibers crossing to the contralateral red nucleus in rats of groups B and C. More BDA-positive fibers crossing the midline and terminating in the contralateral red nucleus in appropriate target areas mirroring the non-differentiated red nucleus were found in rats of group A. Quantitative analysis showed that BDA-labeled axons in the denervated side of rats in group A were more than those in group B （P 〈 0.05）. （2） Improvement in sensorimotor deficit of rats： At 2 weeks after immunization, significant improvement in sensorimotor deficit was found in rats of group A. There were significant differences of improvement in sensorimotor deficit of rats between group A and group B or group C at 12 and 14 weeks after immunization （P 〈 0.05）. CONCLUSION： （1） Poststroke DNA immunization against NGIs leads to increased sensorimotor recovery following FCI and compensatory newly growth of axons from corticorubral projection.     

原料药粒径对盐酸普萘洛尔渗透泵片释药行为的影响

目的:研究原料药粒径等对盐酸普萘洛尔渗透泵片释药行为的影响。方法:取不同批号盐酸普萘洛尔及同批号重结晶前、后的原料药均按相同处方制备成渗透泵片,考察药物体外释放情况及释药24h后衣膜形态;并对上述不同原料药的粒径分别以光学显微镜和激光粒度分析仪进行证实。结果:以原料药粒径较小的渗透泵片释放完毕后衣膜变形,且不能维持零级释放,原料药粒径较大的渗透泵片结果与之相反。不同原料药经仪器证实粒径确有差异。结论:原料药的粒径可影响制备的渗透泵片的释放行为,提示性状稳定的原料药的合理选择在制剂过程中不可忽视。     

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。     

卵巢嵌顿疝致卵巢坏死的危险因素分析

  目的  分析女性婴幼儿卵巢嵌顿性腹股沟斜疝导致卵巢坏死的危险因素。  方法  回顾性研究昆明市儿童医院2018年6月1日至2021年6月1日间,57例女性婴幼儿卵巢嵌顿性腹股沟斜疝（以下简称为卵巢嵌顿疝）并行手术的患儿,分析患儿的年龄、临床资料、实验室检查、术中有无卵巢扭转,并进行单因素及多因素Logistic回归分析。  结果  共纳入57例女性婴幼儿,中位年龄46 d,最小年龄19 d,最大年龄2岁5月。57例患儿均行手术治疗,术中均证实为卵巢嵌顿。单因素分析结果显示白细胞、CRP、卵巢扭转与卵巢坏死有关（P < 0.05）,多因素分析提示卵巢扭转是术中卵巢坏死的独立危险因素（OR = 1.390,95% CI:1.056-1.829,P = 0.019）。  结论  卵巢扭转是婴幼儿卵巢嵌顿性腹股沟疝发生卵巢坏死的独立危险因素。     

ALT升高、低病毒载量慢性乙型肝炎患者不确定期采用不同抗病毒治疗方案的疗效及安全性

  目的  观察初治的低病毒载量不确定期慢乙肝患者采用一线核苷（酸）类药物抗病毒治疗后的临床疗效及安全性。  方法  纳入2019年1月至2022年4月期间在昆明市第三人民医院门诊就诊的96例初治低病毒载量不确定期慢性乙肝患者,根据使用药物不同将患者分为恩替卡韦组（ETV组）、富马酸替诺福韦酯组（TDF组）和富马酸丙酚替诺福韦组（TAF组）。在持续用药的第12周、24周、48周观察3组的临床疗效及安全性差异。  结果  经抗病毒治疗48周后,总共96.88%（93/96）患者HBV DNA获得完全病毒学应答（complete virologic response,CVR,定义为HBV DNA < 100 IU/mL）,3组的CVR率为:ETV组96.97%（32/33）,TDF组96.97%（32/33）,TAF组96.67%（29/30）,组间差异无统计学意义（P = 0.997）。治疗48周时HBsAg水平较基线均有显著下降,但3组差异无统计学意义（P = 0.348）。TAF组较ETV组（P = 0.013）、TDF组（P = 0.047）显著提升eGFR水平,AFP水平较ETV组（P = 0.008）和TDF组（P = 0.007）显著下降。48周ALT复常率:TAF组（70%）和TDF组（45.45%）,ETV组（60.61%）差异无统计学意义（P = 0.135）。治疗期间无严重不良事件发生。  结论  治疗48周后3组CVR率均达96%以上,AFP水平下降及eGFR水平改善TAF组优于ETV组及TDF组,对ALT升高、低病毒载量慢乙肝不确定期患者行抗病毒治疗可使患者获益。