HBcAg对健康人及乙肝病毒携带者外周血单个核细胞刺激产生IFN-α影响的初探

目的观察表达乙肝病毒核心抗原（HBcAg）基因的复制缺陷型重组腺病毒转染外周血单个核细胞后IFN-α产生水平,初步探讨乙型肝炎病毒感染免疫识别状态相关机制。方法应用密度梯度离心法从健康人及乙肝病毒（HBV）携带者外周静脉血中分离出单个核细胞,按分组加入重组腺病毒和（或）CpGODN2006,转染48h后ELISA检测细胞培养上清中IFN-α水平。结果与空白组比较,健康人及携带者各组内IFN-α均有升高,以Ad-HBc＋CpGODN2006组最明显（P〈0.05）,HBV携带者Ad-HBc组IFN-α水平低于健康人（P〈0.05）。结论重组腺病毒感染外周血单个核细胞产生IFN-α水平在乙肝病毒携带者中低于健康人,提示与HBV持续感染造成外周血单个核细胞功能下降有关。     

男性不育患者常规进行MAR法筛查的研究

目的探讨对男性不育患者常规应用混合抗球蛋白反应试验（MAR）筛查精液中抗精子抗体（AsAb）的必要性。方法比较研究组（所有男性不育患者常规应用MAR法筛查抗精子抗体）和对照组（未常规应用MAR法筛查抗精子抗体）两组的抗精子抗体的阳性率。结果对照组抗精子抗体的阳性率是8．6％,研究组抗精子抗体的阳性率是14．3％,P〈0．01。结论男性不育患者常规应用MAR法筛查抗精子抗体可显著提高抗精子抗体的检出率,避免了很多免疫性不育的漏诊。     

TRPC3基因多态性与原发性高血压患者左心室肥厚的关系#br#

目的　探讨TRPC3基因多态性与原发性高血压患者左心室肥厚（LVH）的关系。方法　305例原发性高血压患者根据左心室质量指数（LVMI）分为单纯高血压224例（对照组）和高血压合并LVH 81例（LVH组）。分析2组年龄、吸烟史等一般资料差异。留取外周血标本,采用Sequenom MassARRAY® SNP检测技术对TRPC3基因单核苷酸多态性（SNP）位点Rs2292232、Rs4995894、Rs4292355进行基因分型。Logistic回归分析影响患者LVH的因素。结果　（1）2组年龄、吸烟史、饮酒史、家族史、尿素、肌酐、空腹血糖、三酰甘油、总胆固醇、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇、24 h平均舒张压（24 h DBP）差异无统计学意义（P＞0.05）,LVH组较对照组男性比例降低,而24 h平均收缩压（24 h SBP）、体质量指数（BMI）升高（P＜0.05）。（2）2组Rs4995894及Rs4292355位点等位基因频率、基因型频率分布差异无统计学意义（P＞0.05）,TRPC3基因SNP位点Rs2292232基因型频率和等位基因频率分布差异有统计学意义（P＜0.05）,其中LVH组Rs2292232位点TT基因型频率较对照组更高（P＜0.05）。（3）Logistic回归分析显示,女性（OR=0.332,95%CI：0.181～0.610）、较高24 h SBP（OR=1.035,95%CI：1.014～1.056）、TRPC3基因SNP Rs2292232 TT基因型（OR=2.105,95%CI：1.109～3.995）为原发性高血压患者发生LVH的独立危险因素（均P＜0.05）。结论　TRPC3基因Rs2292232位点SNP与高血压患者发生LVH有关,其中TT基因型患者更易发生LVH。     

增加中期细胞数的染色体标本制备方法研究

目的：探索增加中期细胞数的小鼠骨髓染色体标本制备方法。方法小鼠随机分成2组：对照组与酵母液处理组,每组均采用剪碎股骨法与冲洗股骨法收集骨髓细胞,常规方法制备骨髓染色体标本。结果与对照组比较,酵母液处理组中期细胞数显著增加（P〈0.05）;与冲洗股骨法比较,剪碎股骨法收集的中期细胞数显著增加（P〈0.05）。结论在染色体标本制备过程中,采用酵母液处理和剪碎股骨法,可有效增加小鼠中期细胞数。     

血清肌酸激酶同工酶检测在手足口病诊治中的价值探讨

目的探讨血清肌酸激酶同工酶（CK-MB）检测在手足口病诊治中的价值。方法回顾性分析本院2011年10月至2012年10月收治的手足口病140例,每位患儿均检测了CK-MB,CK-MB≤25U/L72例为A组,CK-MB〉25 U/L68例为B组,对比分析2组的临床特点。结果与A组比较,B组发热峰值明显升高,热程明显延长,呕吐病例、呼吸增快病例、心率增快病例明显增加（P〈0.01）,心肌炎病例增多,中枢神经系统损害病例增多（P〈0.05）,血白细胞平均值明显升高（P〈0.01）,血糖升高病例、ALT升高病例、血乳酸升高病例、心电图异常病例、胸片异常病例明显增多（P〈0.01或P〈0.05）。结论手足口病可以出现心肌损害,合并心肌损害的手足口病患儿病情更严重,常规检测CK-MB有利于对手足口病病情的评估。     

广东汉族人群细胞毒性T淋巴细胞相关抗原-4基因多态性与甲状腺相关性眼病的关系

目的探讨细胞毒性T淋巴细胞相关抗原（CTLA）-4基因外显子1的49位点A／G多态性与甲状腺相关性眼病（TAO）之间的关系。方法研究对象包括TAO患者45例、自身免疫性甲状腺疾病（AITD）无眼病患者100例和本院健康体检中心筛选的无亲缘关系的健康体检者100例。应用聚合酶链反应技术（PCR）对所有血液标本进行CTLA-4基因外显予1的49位点A／G酶切住点限制性片段长度多态性（RFLP）研究，分析比较此多态位点的基因型和等位基因频率在不同人群中分布的差异。结果①TAO组GG基因型频率、G等位基因频率均显著高于无眼病组及对照组（P〈0．05或P〈0．01）；无眼病组GG基因型频率、G等位基因频率亦均显著高于对照组（P〈0．01）。TAO组、无眼病组AA基因型频率均显著低于对照组（P〈0．01），但TAO组与无眼病组相比差异无统计学意义（P=0．118）。②按性别分层后，TAO组、无眼病组男性CG基因型频率及G等位基因频率均显著高于对照组（P〈0．05），而TAO组与无眼病组相比差异无统计学意义（P=0．053）。TAO组、无眼病组女性GG基因型频率及G等位基因频率均同样显著高于对照组（P〈0．05），但TAO组与无眼病组相比亦差异无统计学意义（P=0．075）。③TAO组按甲状腺眼病分级分成5＋6级、4级和3级，3组进行比较发现，3组之间GG基因型频率及G等住基因频率相比差异均无统计学意义（P〉0．05）。结论CTLA-4基因可能是广东地区汉族人群中TAO的易感候选基因。     

原发性肝癌患者体液变化的研究

目的　观察原发性肝癌患者体液分布变化,为患者的营养评价提供依据。方法　选择原发性肝癌患者80例,根据肝功能Child-Pugh分级分为A、B、C组;24例健康体检者作为正常对照（N）组,用生物电阻抗分析法测定人体全身水量（TBW）、细胞外水分（ECW）、细胞内水分（ICW）、浮肿指数（ECW/TBW）、身体细胞量（BCM）。结果　原发性肝癌患者中ICW和BCM不足均占37.5%（30/80）,ECW异常者占42.5%（34/80）;出现水肿（ECW/TBW＞0.39）者占85%（68/80）。B组和C组水肿患者比例明显高于A组（P＜0.05）,其中C组全部患者出现水肿情况;ICW、BCM异常比例随肝功能分级增加而升高（P＜0.05）。C组经过理想体质量（SW）校正后的细胞内水分含量（ICW/SW）低于其他各组（P＜0.05）。各病例组ECW/实际体质量（BW）、ECW/TBW高于N组（P＜0.05）,病例组各组之间差异无统计学意义（P＞0.05）。各病例组患者ICW、ECW、SW校正后的ECW（ECW/SW）、BCM与N组比较差异无统计学意义（P＞0.05）。结论　原发性肝癌患者功能细胞数量进行性消耗,普遍存在细胞内外水分分布异常,应尽早进行合理营养治疗。     

营养支持对肺心病心衰患者治疗的研究

目的：探讨营养支持对肺心病心衰患者治疗的效果。方法：40例肺心病心衰患者随机分为治疗组与对照组,每组20例。两组患者均给予重组人脑钠肽和硝普钠治疗,在此基础上治疗组加用肠内营养支持治疗方法。结果：经过治疗后,两组的PCWP均明显降低（P〈0.05）,LVEF均明显升高（P〈0.05）;与对照组相比,治疗组PCWP下降更明显（P〈0.05）、LVEF升高更明显（P〈0.05）。两组均无严重并发症发生,两组相比差异无统计学意义（P〉0.05）。结论：营养支持对肺心病心衰患者治疗效果好,能显著降低PCWP并升高LVEF,有效改善患者的心功能,值得推广应用。     

雷公藤内酯醇对系统性红斑狼疮患者外周血Th17细胞的影响

目的 观察系统性红斑狼疮（SLE）患者外周血Th17细胞的变化以及雷公藤内酯醇对其的影响。方法 用流式细胞术检测15例SLE稳定期患者、16例SLE活动期患者和32例正常对照者外周血单个核细胞（PBMC）中Th17细胞比例,用RT-PCR检测PBMC中维A酸相关孤儿核受体γt（RORγt）mRNA表达水平。同时对SLE活动期和正常对照组PBMC用雷公藤内酯醇进行干预,培养24h后检测其对Th17细胞比例和RORγt mRNA表达的影响。结果 SLE稳定期及活动期组患者PBMC中Th17细胞比例均高于正常对照组,活动期组患者Th17细胞比例高于稳定期患者;SLE稳定期及活动期组患者PBMC中RORγt mRNA表达水平高于正常对照组,活动期组患者RORγt mRNA表达水平高于稳定期患者,差异均有统计学意义（P〈0.01或P〈0.05）。经雷公藤内酯醇干预后活动期SLE患者PBMC中Th17细胞比例和RORγt mRNA表达水平低于没有干预组,差异有统计学意义（P〈0.05）。结论 SLE患者PBMC中Th17细胞比例和RORγt mRNA表达水平升高,雷公藤内酯醇在体外实验中能降低SLE患者PBMC中Th17细胞比例、减低RORγt mRNA表达水平。     

不同剂量百令胶囊对环孢素A致肾病大鼠免疫功能的影响

目的 观察百令胶囊对环孢素A（cyclosporin A,CsA）大鼠免疫功能的调节作用。方法 SD大鼠,♂,随机分为对照组、模型组、百令大、小剂量组,每组8只。第12周末用流式细胞仪检测大鼠外周血T细胞亚群（CD4^＋、CD8^＋）、B细胞（CD45RA^＋）及NK细胞（CD161a^＋）占淋巴细胞的百分比,计算CD4^＋/CD8^＋比例。ELISA法测定外周血血清中IgG,IgM含量。结果 模型组及百令大、小剂量组大鼠外周血中CD4^＋与CD8^＋百分比均稍高于对照组,但差异无统计学意义（P〉0.05）。模型组CD4^＋/CD8^＋比值显著低于对照组（P〈0.05）,百令大、小剂量组CD4^＋/CD8^＋比值显著高于模型组（P〈0.05）,百令大剂量组CD4^＋/CD8^＋比值显著高于百令小剂量组（P〈0.05）,其余各组间比较差异无统计学意义。与对照组比较,模型组外周B细胞占淋巴细胞的百分比及血清IgG、IgM含量显著降低（P〈0.05或P〈0.01）;与模型组比较,百令大、小剂量组外周B细胞占淋巴细胞的百分比及血清IgG、IgM含量显著升高（P〈0.05,P〈0.01）;百令大剂量组B细胞（CD3^－CD45RA^＋）比值显著高于百令小剂量组（P〈0.05）。结论 百令胶囊对CsA所致肾病大鼠免疫失调及低下具有调节和促进作用,大剂量百令胶囊作用更明显。     

Induction of meiotic micronuclei in spermatocytes in vivo by aflatoxin B1: light and transmission electron microscopic study in Swiss mouse

Dietary aflatoxins produce a disease state known as aflatoxicosis, and disruption of spermatogenesis is one of its serious consequences. Towards finding the cellular targets in spermatogenic compartment for aflatoxin toxicity, aflatoxin B(1) (AFB(1)) was administered to 90-day-old Swiss mouse through i.p. route at a daily dose of 20mug per kg body weight for 7, 15, 35 and 45 days. The testis and epididymis were subjected to light- as well as transmission electron microscopic analysis. One of the newer observations was occurrence of meiotic micronucleate giant spermatocytes in seminiferous epithelium and epididymal lumen. The origin of these cells could be traced to imminent disruption of spindle apparatus during meiotic division of spermatocytes, resulting in lagging of chromosome bivalents or replicated univalents. Such chromosomes appeared to undergo condensation and become micronuclei. Thus, this study reports that aflatoxin exposure would result in generation of meiotic micronucleate giant spermatocytes.     

Induction of meiotic micronuclei in spermatocytes in vivo by aflatoxin B1: Light and transmission electron microscopic study in Swiss mouse

Dietary aflatoxins produce a disease state known as aflatoxicosis, and disruption of spermatogenesis is one of its serious consequences. Towards finding the cellular targets in spermatogenic compartment for aflatoxin toxicity, aflatoxin B₁ (AFB₁) was administered to 90-day-old Swiss mouse through i.p. route at a daily dose of 20 μg per kg body weight for 7, 15, 35 and 45 days. The testis and epididymis were subjected to light- as well as transmission electron microscopic analysis. One of the newer observations was occurrence of meiotic micronucleate giant spermatocytes in seminiferous epithelium and epididymal lumen. The origin of these cells could be traced to imminent disruption of spindle apparatus during meiotic division of spermatocytes, resulting in lagging of chromosome bivalents or replicated univalents. Such chromosomes appeared to undergo condensation and become micronuclei. Thus, this study reports that aflatoxin exposure would result in generation of meiotic micronucleate giant spermatocytes.     

六种抗肿瘤药物影响小鼠精子生成过程的定量组织学比较

目的通过对小鼠睾丸的定量组织学研究,观察丝裂霉素C等6种抗肿瘤药物对小鼠睾丸生精过程的影响。方法制作小鼠睾丸组织切片,显微镜下观察,记录第Ⅶ相细精管的精原细胞数及第Ⅳ相细精管的次级精母细胞和处于减数分裂中期、后期的细胞数。结果腹腔注射给药,丝裂霉素C和美法仑可使小鼠睾丸精原细胞接近完全消失,而多柔比星、长春新碱、甲氨蝶呤和5-氟尿嘧啶对精原细胞没有影响。此6种抗肿瘤药物对次级精母细胞以及处于减数分裂中期、后期的细胞均无明显影响。给小鼠腹腔注射丝裂霉素C2mg.kg-1,3d后第Ⅶ相细精管的精原细胞接近完全消失,14d后精母细胞接近完全消失,21d后精子细胞消失,35d后3种细胞数目基本恢复正常。结论不同抗肿瘤药物对小鼠睾丸精原细胞的影响有明显的差异。精原细胞对丝裂霉素C高度敏感。6种抗肿瘤药物对次级精母细胞和处于减数分裂中期、后期的细胞数量无明显影响。     

Evolution of DNA strand-breaks in cultured spermatocytes: the Comet Assay reveals differences in normal and gamma-irradiated germ cells.

In reproductive toxicity assessment, in vitro systems can be used to determine mechanisms of action of toxicants. However, they generally investigate the immediate effects of toxicants, on isolated germ cells or spermatozoa. We report here the usefulness of in vitro cultures of rat spermatocytes and Sertoli cells, in conjunction with the Comet Assay to analyze the evolution of DNA strand-breaks and thus to determine DNA damage in germ cells. We compared cultures of normal and gamma-irradiated germ cells. In non-irradiated spermatocytes, the Comet Assay revealed the presence of DNA strand-breaks, which numbers decreased with the duration of the culture, suggesting the involvement of DNA repair mechanisms related to the meiotic recombination. In irradiated cells, the evolution of DNA strand-breaks was strongly modified. Thus our model is able to detect genotoxic lesions and/or DNA repair impairment in cultured spermatocytes. We propose this model as an in vitro tool for the study of genotoxic injuries on spermatocytes.     

Comparative testicular toxicity of bis(2-methoxyethyl) ether and 2-methoxyethanol in rats

Male rats were exposed to 0, 110, 370, or 1100 ppm bis(2-methoxyethyl)ether (diglyme) 6 h/day, 5 days/week for 2 weeks. One group of male rats was exposed to 300 ppm 2-methoxyethanol (2-ME) for 2 weeks as a positive control. Exposed rats were killed after 10 days of exposure and 14, 42, or 84 days post-exposure (PE), respectively. At 110 ppm diglyme, spermatocytes in pachytene and meiotic division at spermatogenic stages XII-XIV were mainly affected. At 370 ppm diglyme, affected germ cells were similar to those seen at 110 ppm diglyme, but round spermatids at spermatogenic stages I-VII were also affected. The testes regained normal spermatogenesis by 84 days PE. At 1100 ppm diglyme or 300 ppm 2-ME, marked testicular atrophy was found affecting all spermatogenic stages. Damaged seminiferous tubules were lined with regenerating pachytene spermatocytes at 14 days PE and with spermatocytes and round spermatids after 42 days PE. Most but not all testes in rats exposed to 300 ppm 2-ME or 1100 ppm diglyme had normal morphology after 84 days PE. Based on the observation of germ cell damage, spermatozoa population in the epidymal tubules, reversibility of spermatogenesis after various PE periods, testicular toxicity induced by 300 ppm 2-ME was more severe than that seen at 370 ppm diglyme but was slightly less remarkable than that of 1100 ppm diglyme.     

Unscheduled DNA synthesis in the testis,a secondary test for the evaluation of chemical mutagens

DNA damage induced by methylmethane sulfonate, cyclophosphamide, doxorubicin and procarbazine in male germ cells was assessed in rabbits by the demonstration of unscheduled DNA synthesis (UDS), in meiotic and postmeiotic phases of maturation. Immediately after treatment by the intravenous route tritiated thymidine was injected into both testicles. Subsequently, rabbits were ejaculated serially, sperm heads were isolated and assayed for radioactivity by liquid scintillation counting. Dose-dependent UDS was demonstrated in late spermatocytes and early spermatids. High doses of hycanthone also induced UDS, but isoniazid and metronidazole had no effect. The rabbit testis UDS test takes into account metabolic and pharmacokinetic aspects of the test substances and provides information about their penetration through the blood-testicular barrier. It is therefore useful for secondary evaluation of potential mutagens. UDS induced by procarbazine was abolished by simultaneous treatment with Ara-C. Thus, the test also recognizes substances that inhibit DNA repair synthesis.     

Comparison of the in vivo and in vitro testicular effects produced by methoxy-, ethoxy- and N-butoxy acetic acids in the rat

Methoxy-, ethoxy- and n-butoxy acetic acids, known urinary metabolites of the corresponding alkoxyethanol solvents, were administered by gavage to rats as a single oral dose equimolar with 500, 250 or 100 mg 2-methoxyethanol/kg body weight. Testicular weight and morphology were monitored over a 14-day period post-treatment. Methoxyacetic acid (MAA) was the only compound which produced a significant decrease in testicular weight. Histological examination of the testes from treated animals indicated that MAA at all doses and ethoxyacetic acid (EAA) at the highest dose, produced damage specific to spermatocytes undergoing meiotic maturation and division (particularly stages XIII-XIV) within 24 h of treatment. n-Butoxyacetic acid (BAA) had no discernable effect on the testis at any dose level or time, although there was evidence of haematuria produced by the compound. Addition of MAA, EAA and BAA to testicular cell cultures at concentrations approximately equivalent to the steady state plasma levels of MAA determined after a testicular toxic dose (500 mg/kg) of methoxyethanol (5 mM) produced a specific loss of pachytene spermatocytes (the target population in vivo) from the system by MAA and EAA (MAA greater than EAA). BAA did not produce any specific changes to testicular cell populations in vitro. Thus the production of testicular toxicity by alkoxyacetic acids diminishes with increasing chain length, and a good correlation exists between in vivo and the in vitro system.     

Apoptosis in male germ cells in response to cyclin A1-deficiency and cell cycle arrest

Male mice homozygous for a mutated allele of the cyclin A1 gene (Ccna1) are sterile due to a block in cell cycle progression before the first meiotic division. Meiosis arrest in Ccna1(-/-) spermatocytes is associated with desynapsis abnormalities, lowered MPF activity, and apoptosis as evidenced by TUNEL-positive staining. With time, adult testicular tubules exhibit severe degeneration: some tubules in the older animals are almost devoid of germ cells at various stages of spermatogenesis. The mechanisms by which the cells sense the cell cycle arrest and the regulation of the decision to undergo cell death are under investigation.     

Testicular toxicity of rats exposed to hexafluoroacetone (HFA) for 90 days

Rats were exposed to 0, 0.1, 1, and 12 ppm of hexafluoroacetone (HFA) for 6 h/day, 5 days/week for 90 days. The exposed rats were killed after 30 or 90 days exposure, and 28 or 84 days post-exposure (PE). There were no exposure-related pathological lesions in the rats exposed to 0.1 or 1.0 HFA for 90 days. After 30 days exposure to 12 ppm HFA, rats showed lower body weight gain, testicular atrophy, and oligospermia or aspermia in the epididymal tubules. At 30 days exposure, the atrophic testes had marked depletion of round spermatids in spermatogenic stages I-VIII and elongated spermatids in spermatogenic stages IX-XIV, but mature spermatids appeared only slightly decreased. Numerous spermatocytes in meiotic division in spermatogenic stage XIV were necrotic. At 90 days exposure, the testes showed severe atrophy with almost all seminiferous tubules affected and both immature and mature spermatids had disappeared from the seminiferous tubules. The epididymal tubules were devoid of spermatozoa. After 28 days PE, regeneration of atrophic testes was evident but varied markedly among the exposed rats. The number of seminiferous tubules producing elongated and mature spermatids was significantly lower than that of normal testes. Many seminiferous tubules had not regained normal spermatogenesis and the epididymal tubules showed marked oligospermia. After 84 days PE, normal spermatogenesis was still only partially restored to the atrophic testes, with many of the regenerating tubules still devoid of normal spermatogenesis.     

The Recovery of the Testis over 8 Weeks after Short-Term Dosing with Ethylene Glycol Monomethyl Ether: Histology, Cell-Specific Enzymes, and Rete Testis Fluid Protein

The Recovery of the Testis over 8 Weeks after Short-Term Dosingwith Ethylene Glycol Monomethyl Ether Histology, Cell-SpecificEnzymes, and Rete Testis Fluid Protein. CHAPIN, R. E., DUTTON,S. L., ROSS, M. D., SWAISGOOD, R. R., AND LAMB, J. C, IV (1985)Fundam. Appl. Toxicol. 5, 515–525. Ethylene glycol monomethylether (EGME) has been found to affect meiotic spermatocytes,spermatids, other stages of spermatocytes, and spermatogonia,depending on the dose used. These studies, which examine testicularrecovery from EGME treatment, analyzed tissues from rats treatedfor 5 days with 0, 50, 100, or 200 mg EGME/kg/day and sacrificedat eight subsequent weekly intervals; some epididymal spermparameters of these animals have been described. Histologically,the testes of the low-dose group showed very mild changes, whilethe 100- and 200-mg/kg groups showed widespread damage and celldeath which recovered somewhat during the course of the study.There was no treatment-related effect on seminal vesicle orprostate weights. Rete testis fluid protein levels were changedonly in the high-dose group, when protein levels rose to a maximumof sixfold the control values at Week 4; by Week 6, there wasno difference between groups. Changes in cell-specific enzymeactivities were dose dependent and generally mirrored changesin the number of germ cells in the testis