柱切换HPLC法测定腹水中左旋氧氟沙星的含量

目的：应用柱切换ＨＰＬＣ技术直接测定腹水中左旋氧氟沙得含量。方法：采用磷酸二氢钾缓冲溶液（ＰＨ２．２）－０．０５ｍｏｌ／Ｌ四丁溴化化铵（１００：４）为预处理流动相,经μＢｏｎｄａｐａｋ Ｃ１８柱预处理后,切换到Ｉｒｒｅｇｕｌａｒ＿ＨＣ１８分析柱,以甲醇－磷权二氢钾缓冲溶液（ＰＨ２．２）－０．０５ｍｏｌ／Ｌ四丁基溴化铵（３０：７０：４）为分析流动相进行分离测定,紫外线皮长为２９４ｎｍ。结果：左旋氧氟     

一种新的融合表达载体的构建

利用Ｌ－ａｎｓＢ基因片段构建了融合表达载体ｐＥＣ,其中Ｌ－ａｎｓＢ基因中编码唯一酸水解位点的序列已通过定点突变消除,并在它的３’端引入了新的酸水解位点编码序列和方便外源基因插入的克隆位点ＢａｍＨＩ和ＨｉｎｄⅢ,外源基因可从克隆位点ＢａｍＨＩ和ＨｉｎｄⅢ处插入ｐＥＣ中,表达的融合蛋白用酸水解法加工后仅被切割成一大一小两个片段,不会导致产物混杂,ｈＧＲＦ对它进行验 结果显示ｈＧＲＦ基因不仅能按正确的读     

天津市1999-2021年0~14岁儿童伤害死亡谱及城乡差异分析

目的 分析天津市0~14岁儿童伤害死亡谱的特征、变化情况及城乡差异。方法 1999-2021年天津市儿童伤害死亡数据来源于天津市全人口全死因监测数据库,计算不同亚组人群和主要伤害原因的构成比、粗死亡率和标化死亡率并比较城乡差异。采用Cochran-Armitage趋势检验分析死亡原因构成比的时间变化趋势。采用Joinpoint回归分析变化趋势,计算平均年变化百分比（AAPC）。伤害死亡风险的季节差异用死亡率比值及其95%CI表示。结果 1999-2021年,伤害是天津市0~14岁儿童的第3位死因。农村儿童死于医疗卫生机构的比例为31.08%,低于城市的37.82%。儿童伤害的总体标化死亡率呈下降趋势（AAPC=-5.54%,P<0.001）。溺水和道路交通伤害的标化死亡率在城市和农村地区呈下降趋势（P<0.001）。意外中毒的标化死亡率仅在农村地区呈下降趋势（AAPC=-8.09%,P<0.001）,在城市地区无明显变化趋势（P>0.05）。自杀标化死亡率在城市地区无明显变化趋势（P>0.05）,在10~14岁农村儿童中呈上升趋势（AAPC=4.58%）。跌倒/坠落标化死亡率在城市和农村地区均无明显变化趋势（P>0.05）。伤害死亡的总体风险和溺水死亡风险在城乡均为夏季最高;道路交通伤害在城市为秋季最高,在农村为夏季最高;意外中毒死亡风险在城乡均为冬季最高。结论 近年来天津市儿童伤害死亡情况得到明显改善。城市和农村地区的儿童伤害死亡水平仍存在较大差异,在未来政策制定中,应充分考虑缩小城乡差距。     

Gene cloning and sequencing of BmK AS and BmK AS-1, two novel neurotoxins from the scorpion Buthus martensi Karsch.

Based on the known amino acid sequences of BmK AS and BmK AS-1, the gene specific primers were designed and synthesized for 3' and 5' RACE (Rapid Amplification of cDNA Ends). Their partial cDNA fragments obtained by 3' and 5' RACE were cloned and sequenced, and the full length cDNA sequences of BmK AS and BmK AS-1 were then completed by overlapping their two partial cDNA sequences, respectively. The predicted amino acid sequences both consist of 85 amino acid residues including a putative signal peptide of 19 residues and a mature toxin of 66 residues. They are different in 17 amino acid residues, among them 11 residues in the mature toxin. The predicted amino acid sequences of BmK AS and BmK AS-1 were almost consistent with those determined and revised (personal communication), only different in one and two residues at their COO-terminal parts, respectively. Based on the determined cDNA sequences, and using the total DNAs isolated from the scorpion venom glands as a template, the genomic DNAs of BmK AS and BmK AS-1 were also amplified by PCR and sequenced. It showed that no intron was inserted in their open reading frames, while in the exon of signal peptide sequences of other Na+, K+ and Cl- channel toxins from the same scorpion, an intron is usually found. However, the Northern blot hybridization results indicated that the sizes of their mRNA should be around 800 bp. Their extra sequences around 400 bp which might function as an intron should be located at their 5' untranslated regions.     

Characterization of descending facilitation and inhibition of spinal nociceptive transmission from the nuclei reticularis gigantocellularis and gigantocellularis pars alpha in the rat.

1. Descending influences produced by focal electrical stimulation in the nuclei reticularis gigantocellularis (NGC) and gigantocellularis pars alpha (NGC alpha) on spinal nociceptive transmission and the dorsoventral region of spinal white matter mediating stimulation-produced modulation were examined in pentobarbital sodium-anesthetized, paralyzed rats. Spinal units studied responded to mechanical stimuli and noxious heating (50 degrees C) of cutaneous receptive fields confined to the glabrous skin of the ipsilateral hind foot. Recording sites were located in laminae I-VI of the L3-L5 spinal segments. 2. Electrical stimulation in the NGC or NGC alpha produced both facilitation and inhibition of responses of spinal units to noxious heating of the skin. At 33 of 57 stimulation sites in the NGC and NGC alpha, electrical stimulation produced biphasic effects, facilitating responses at lesser intensities (5-25 microA) and inhibiting responses at greater intensities (50-100 microA). At 21 other sites in the NGC and NGC alpha, electrical stimulation (5-100 microA) only inhibited, and at 3 sites stimulation (5-100 microA) only facilitated responses of spinal units to noxious heating of the skin. 3. Electrical stimulation in the NGC or NGC alpha contralateral to the spinal recording site produced the same magnitude of facilitation/inhibition or inhibition of spinal nociceptive transmission as did stimulation in the ipsilateral NGC and NGC alpha. 4. The latencies to descending facilitation and inhibition of spinal nociceptive transmission from the NGC and NGC alpha were estimated by a cumulative sum technique to be 232 and 80 ms, respectively. 5. Responses of spinal units to graded heating (42-50 degrees C) of the skin exhibited positively accelerating stimulus-response functions (SRF) throughout the temperature range tested. Electrical stimulation at lesser, "facilitating" intensities produced a parallel, leftward shift of the SRF, whereas stimulation at greater, "inhibitory" intensities significantly decreased the slope of the SRF without affecting the threshold for response. 6. To determine whether activation of cell bodies in the NGC or NGC alpha were capable of replicating the effects of electrical stimulation, L-glutamate was microinjected into sites where electrical stimulation facilitated at lesser and inhibited at greater intensities the responses of spinal units to 50 degrees C heating of the skin. L-Glutamate (5 nmol) produced a rapid onset, short-lasting and reproducible facilitation of nociceptive transmission; glutamate microinjection into the same site at a greater dosage (50 nmol) inhibited responses of the same spinal units.(ABSTRACT TRUNCATED AT 400 WORDS)     

消旋甲磺酸帕珠沙星的合成

目的：合成消旋甲磺酸帕珠沙星。方法：以市售合成氧氟沙星的中间体9，10-二氟-3-甲基-7-氧-2，3-二氢-7H-吡啶[1，2，3-d，e][1，4]苯并噁嗪-6-羧酸乙酯为起始原料，经过缩合反应、水解脱羧、相转移缩-环合、腈基水解和Hofmann降解得目标化合物。结果和结论：经^1H-NMR，MS和旋光活性测定，证明所得到的产物为消旋甲磺酸帕珠沙星。     

E试验监测重症监护病房中400株革兰氏阴性杆菌的耐药性

目的 调查我院监护病房革兰氏阴性杆菌耐药状况及超广谱β—内酰胺酶(ESBLs)菌株的发生率，了解四年来革兰氏阴性菌对抗菌药物的耐药性变迁。方法 用E试验法测定400株革兰氏阴性杆菌对11种抗生素的最低抑菌浓度(MIC)。并用头孢他啶／头孢他啶 克拉维酸E试验酶试条检阅细菌产ESBL的情况。结果 亚胺培南对所有受试菌保持最高抗菌活性，细菌耐药率仅为7．5％；头孢哌酮／舒巴坦、阿米卡星、哌拉西林／三唑巴坦、头孢吡肟和头孢他啶仍有较高抗菌活性，耐药率分别为16．0％、20．0％、22．5％、23％和26．8％；其它抗生素耐药率在42．8％—46.8％之间。用头孢他啶／头孢他啶 克拉维酸筛选大肠埃希氏菌和肺炎克雷伯氏菌ESBLs产生菌27株，检出率分别为40．9％和27．3％。体外抗菌活性最强的是亚胺培南，所有大肠埃希氏菌和肺炎克雷伯氏菌ESBLs菌株都对之敏感。含酶抑制剂抗生素哌拉西林／三唑巴坦和头孢哌酮／舒巴坦对产ESBLs株保持较高体外抗菌活性，但2001年分离菌耐药率有所增加。四年资料相比，所测细菌对头孢哌酮／舒巴坦耐药率由1998—2000年的11％—14％增加至2001年的26％，大肠埃希氏菌对环丙沙星耐药率由1998—1999年的45.2％上升至2000—2001年的79.6％，细菌对其它抗菌药物耐药率变化不大。结论 亚胺培南抑菌率最高，对ESBLs产生菌有较强的抗菌活性，而抗菌药物(尤其是加酶抑制剂抗生素和喹诺酮类药物)的滥用，已造成细菌对其耐药率升高，且对付产酶耐药株感染逐渐失去优势。     

院内感染嗜麦芽寡养单胞菌耐药性分析

目的 测定26株临床分离院内感染嗜麦芽寡养单胞菌对常用抗菌药物的耐药性，探索产β-内酰胺酶菌株对亚胺培南的耐药机制。方法 采用琼脂二倍稀释法测定19种抗菌药物对26株临床分离的院内感染嗜麦芽寡养单胞菌的体外抗菌活性，并比较产酶株与非产酶株对抗菌药物的敏感性；测定产酶株所产肛内酰胺酶对亚胺培南的稳定性和丝氨酸β-内酰胺酶、金属β-内酰胺酶抑制剂的抑酶保护效应。结果 新型氟喹诺酮类药物司帕沙星、左氧氟沙星、加替沙星和多西环素对26株嗜麦芽寡养单胞菌有较强抗菌活性。抑菌率分别为96．2％、96．2％、92．3％、84．6％，SMZ／TMP和替卡西林／克拉维酸抑菌率分别为65．4％和50％，亚胺培南、头孢他啶、头孢哌酮和氨曲南的敏感性较差，耐药率在80％以上；产酶株与非产酶株药敏结果比较，产酶株对含酶抑制剂β-内酰胺类抗生素的敏感性较非产酶株高，而对其它抗菌药物的敏感性变化不大。酶稳定性试验显示，有4株产酶菌产碳青霉烯水解酶，酶抑制试验表明，克拉维酸对这4株菌所产酶无抑酶保护效应，而金属β-内酰胺酶抑制剂EDTA对4株菌所产酶有抑制作用，且酶活性能被ZnCl2复活。结论 新型氟喹诺酮类药物对嗜麦芽寡养单胞菌院内感染株有较强的抗菌活性；4株产酶菌所产碳青霉烯水解酶可能为金属β-内酰胺酶。     

皮肤过敏原点刺试验在儿童哮喘病原诊断中的应用

目的 了解皮肤过敏原点刺试验在哮喘病原诊断中的应用。方法 变应原皮试液由上海医科大学提供，用生理盐水作阴性对照，盐酸组织胺作阳性对照。以患儿前臂屈侧，婴幼儿亦可用背部皮肤进行点刺试验。结果 阳性结果依次为上海螨、屋尘、棉絮、蟑螂、枕垫、霉菌和花粉等，全部阴性20例。结论 从一个侧面说明了本地区引起哮喘发作的各种致敏因素，皮肤过敏原点剌试验对患儿痛苦小，易于操作，重复性较高，但尚存在假阳性及假阴性的情况，操作时针刺点深浅掌握不好易影响结果，故对其结果必须结合临床具体分析。     

复方毛冬青注射液与3种输液配伍后的微粒及其pH值观察

目的　观察复方毛冬青注射液分别与 0 9%氯化钠注射液、10 %葡萄糖注射液和小儿用葡萄糖氯化钠注射液 3种输液配伍时的不溶性微粒及其pH值变化。方法　不同剂量 ( 4、6、8和 10ml)的复方毛冬青注射液分别与 10 0ml上述输液配伍 ,混合液的不溶性微粒利用微粒分析仪测定 ,以及其 pH值利用pH/离子计仪测定。 结果　不同剂量复方毛冬青注射液与各输液混合其 4个通道 (≥ 2 5 μm、≥ 10 μm、≥ 5 μm、≥ 2 μm)的不溶性微粒均显著性增多 (P <0 .0 1) ,且其pH也有明显变化。 结论　复方毛冬青注射液与 3种输液配伍的不溶性微粒不符合中国药典及英国药典的质量标准 ,应引起重视。建议在临床应用时可在其一次性输液器加装有效的终端滤器