精子特异性乳酸脱氢酶研究进展

特异性乳酸脱氢酶( sperm-specific lactatedehydrogenase,LDH-C4)特异地存在于鸟类和哺乳类动物的睾丸和精子中,与体细胞乳酸脱氢酶A4(LDH-A4)和B4(LDH-B4)同属于乳酸脱氢酶家族,它们都以烟酰胺腺嘌呤二核苷酸(NAD)为辅酶催化丙酮酸和乳酸的转化,在能量代谢中发挥重要作用,但LDH-C4 与体细胞LDH 相比又具有一些独特性质.近来,在研制新型避孕药的过程中,免疫避孕疫苗逐渐引起人们的重视[1],免疫法控制生育具有无药物的毒副作用、管理使用方便、低价、作用相对持久且可逆等优势,但找到理想的精子特异性抗原是制备免疫避孕疫苗的关键[2].用不育患者血清中提取的抗精子抗体(AsAb)来寻找睾丸cDNA 文库表达的抗原候选对象,结果发现特征性强且免疫效果可靠的免疫原为LDH-C4[3].本文主要针对近年来LDH-C4 的研究进展及其应用做一综述.

Objective A diagnostic model was established to discriminate infectious diseases from non-infectious diseases. Methods The clinical data of patients with fever of unknown origin (FUO) hospitalized in Xiangya Hospital Central South University, from January, 2006 to April, 2011 were retrospectively analyzed. Patients enrolled were divided into two groups. The ifrst group was used to develop a diagnostic model: independent variables were recorded and considered in a logistic regression analysis to identify infectious and non-infectious diseases (αin= 0.05, αout= 0.10). The second group was used to evaluate the diagnostic model and make ROC analysis. Results The diagnostic rate of 143 patients in the ifrst group was 87.4%, the diagnosis included infectious disease (52.4%), connective tissue diseases (16.8%), neoplastic disease (16.1%) and miscellaneous (2.1%). The diagnostic rate of 168 patients in the second group was 88.4%, and the diagnosis was similar to the ifrst group. Logistic regression analysis showed that decreased white blood cell count (WBC < 4.0×109/L), higher lactate dehydrogenase level (LDH > 320 U/L) and lymphadenectasis were independent risk factors associated with non-infectious diseases. The odds ratios were 14.74, 5.84 and 5.11 (P≤ 0.01) , respectively. In ROC analysis, the sensitivity and speciifcity of the positive predictive values was 62.1% and 89.1%, respectively, while that of negative predicting values were 75% and 81.7%, respectively (AUC = 0.76,P = 0.00). Conclusions The combination of WBC < 4.0×109/L, LDH > 320 U/L and lymphadenectasis may be useful in discriminating infectious diseases from non-infectious diseases in patients hospitalized as FUO.

In this study, we hypothesized that total flavonoid of Litsea coreana leve (TFLC) protects against focal cerebral ischemia/reperfusion injury. TFLC (25, 50, 100 mg/kg) was administered oral y to a rat model of focal ischemia/reperfusion injury, while the free radical scavenging agent, edaravone, was used as a positive control drug. Results of neurological deficit scoring, 2,3,5-triphenyl tetrazolium chloride staining, hematoxylin-eosin staining and biochemical tests showed that TFLC at different doses significantly al eviated cerebral ischemia-induced neurological deficits and histopathological changes, and reduced infarct volume. Moreover, it suppressed the increase in the levels of nitrates plus nitrites, malondialdehyde and lactate dehydrogenase, and it diminished the reduction in gluta-thione, superoxide dismutase and catalase activities induced by cerebral ischemia/reperfusion in-jury. Compared with edaravone, the protective effects of TFLC at low and medium doses (25, 50 mg/kg) against cerebral ischemia/reperfusion injury were weaker, while the protective effects at high dose (100 mg/kg) were similar. Our experimental findings suggest that TFLC exerts neuroprotective effects against focal cerebral ischemia/reperfusion injury in rats, and that the effects may be asso-ciated with its antioxidant activities.

Baicalin, a type of flavonoid extracted from the dried root of Scutel aria baicalensis georgi, has been shown to effectively inhibit cellapoptosis. Therefore, we assumed that baicalin would suppress colistin sulfate-induced neuronal apoptosis. PC12 cells exposed to colistin sulfate (62.5-500 μg/mL) for 24 hours resulted in PC12 cellapoptosis. In addition, caspase-3 activity, lactate dehydrogenase level and free radical content increased in a dose-dependent manner. Subsequently, PC12 cells were pretreated with baicalin (25, 50 and 100 μg/mL), and exposed to 125 μg/mL colistin sulfate. cellmorphology markedly changed, and cellviability increased. Moreover, caspase-3 activity, lac-tate dehydrogenase level and free radical content decreased. Results indicated that baicalin inhib-ited colistin sulfate-induced PC12 cellapoptosis by suppressing free radical injury, and reducing caspase-3 activity and lactate dehydrogenase activity.

Objective. The present study investigated the role of calcineurin in angiotensin II(AngII) induced cardiac myocyte hypertrophy of rats. Method. The primary cardiac myocytes were cultured under the standard conditions. The calcineurin activity in AngII treated cardiomyocytes was tested by using PNPP;protein synethsis rate was assessed by 3H leucine incorporation; atrial natriuretic factor(ANF) Mrna level was determined by Northern blot analysis. Cell viability was estimated by lactate dehydrogenase(LDH) levels in cultured medium and by dyed cell numbers. Result. After stimulation of 10,100 and 1 000nmol/L of AngII, calcineurin activities in the cardiomyocytes were increased by 13％ ,57％ (P< 0.05) and 228％ (P< 0.01) respectively, compared with control group. Cyclosporin A(CsA), a specific inhibitor of calcineurin, markedly inhibited the calcineurin activity and decreased the 3H leucine incorporation in AngII treated cardiomyocytes in a dose dependent manner. It was also found that CsA slightly reduced the Mrna level of ANF gene in AngII stimulated cardiomyocytes. Conclusion. During AngII induced cardiac myocyte hypertrophy, calcineurin signal pathway is activated, and inhibition of the pathway can attenuate AngII induced cardiac myocyte hypertrophy, which suggests that the calcineurin signal pathway may play an important role in AngII induced myocardial hypertrophy of rats.

早期颅脑损伤患者空腹血糖及乳酸脱氢酶表达的研究

随着工农业及交通运输业的发展,重大创伤事故发生率逐年提高,在创伤中颅脑损伤患者急剧增加,患者的致死率、致残率很高.神经外科医师在颅脑损伤早期准确判断损伤的轻重,并给予适当的救治尤其重要.本研究检测65例颅脑损伤患者在急性期时空腹血糖及乳酸脱氢酶的含量,旨在为判断颅脑损伤的预后提供必要的理论支持.

泌尿外科常用术语英文缩写(之三)

(续第15卷第9期第433页)IA idiopathic Addison＇s disease特发性阿狄森氏病IAH idiopathic adrenal hyperplasia特发性肾上腺增生IBI intermittent bladder irrigation间歇性膀胱灌注法IC ion chromatography离子色谱(法)ICT immunoreactive calcitonin免疫反应性降钙素ID idiotype个体基因型IEC intraepithelial carcinoma上皮内癌IF intrinsic factor内因子Ig immunoglobulin免疫球蛋白IHC idiopathic hypercalcemia特发性高钙血IHC idiopathic hypercalciuria特发性高钙尿IL-2 interleukin-2白细胞介素2IMP incomplete male pseudohermaphroditism不完全性男性假两性畸形INF interferon干扰素IP intravenous pyelography静脉肾盂造影术IPARF idiopathic postpartum acute renal failure特发性产后急性肾功能衰竭IS index of sexuality性指数ITFS incomplete testicular feminization syndrome不完全性睾丸女性化综合征IVF in vitro fertilization体外受精IVP intravenous pyelography静脉(或排泄性)肾盂造影术IVU intravenous urography静脉尿路造影术K17-K 17-ketosteroid17-酮类固醇KFAB kidney-fixing antibody肾结合抗体KGT kidney glucosidetransferase肾葡萄糖苷转移酶KUB kidneys,ureters,bladder肾，输尿管，膀胱LLAD lactic acid dehydrogenase乳酸脱氢酶LAF lymphocyte activating factor淋巴细胞活化因子LAG lymphangiogram淋巴管造影照片LASER light amplification by stimulated emission of ra-diation (Laser)激光LC liquid chromatography液相色谱(法LDH lactate dehydrogenase乳酸脱氢酶LGV lymphogranuloma venereum性病性淋巴肉芽肿LH luteinizing hormone促黄体(生成)激素LK left kidney左肾LU left ureter左输尿管Lues I primary syphilis一期梅毒Lys lysine赖氨酸MM male男性，雄性MA membrane antigen膜抗原MC mineralocorticoid(肾上腺)盐皮质激素MMC mitomycin丝裂霉素McAb monoclonal antibody单克隆抗体MDR multidrug resistance多药耐药MFR maximum flow rate大流率MGDS ixed gonadal dysgenesis syndrome混合型性腺发育不良综合征MHC major histocompability complex主要组织相容性(抗原)复合物MLD minimal lethal dose小致死量MLD50 median lethal dose半数致死量MNT 17-α-methyl-19-nortestosterone甲基去甲睾丸酮MoAb monoclonal antibody单克隆抗体MOF multiple organ failure多器官衰竭MP mucopolysaccharide粘多糖MPO maintenance peritoneal dialysis维持腹膜透析MPSI male procreative superiority index男性生殖优势指数MRI magnetic resonance imaging磁共振成象mRNA messenger RNA ribonucleic acid信使核糖核酸MSOF multiple system organ failure多系统器官衰竭MSSU mid-stream specimen of urine中段尿标本MSU monosodium urate单钠尿酸盐(尿酸一钠)MT Malpighian tubule马尔皮吉安氏小管(肾小管)MT methyl testosterone甲基睾丸素MTS multiple tumor suppressor多肿瘤抑制因子NNDI nephrogenic diabetes insipidus肾性尿崩症NGU nongonococcal urethritis非淋菌性尿道炎NO nitric oxide一氧化氮NOS nitric oxide synthase一氧化氮合成酶NS nephrotic syndrome肾病综合征NS normal saline生理盐水NTAB nephrotoxic antibody肾毒性抗体O17-OHCS 17-hydroxycorticosteroids17-羟皮质类固醇25-OHD 25-hydroxyvitamin D25-羟维生素D25-OHDa 25-hydroxycholecalciferol25-羟维生素D31,25-(OH)2D3 1,25-dihydroxycholecalciferol1，25-二羟胆钙化醇Orn ornithine鸟氨酸 (待续)(王少刚叶章群整理)

AIM:Programmed necrosis ( necroptosis ) and apoptosis are crucially involved in multiple severe cardiac pathological conditions , including myocardial infarction, ischemia/reperfusion (I/R) injury, and heart failure.Whereas apoptotic signaling is well defined, the mechanisms underlying cardiomyocyte necroptosis remain elusive .METHODS AND RESULTS:Here we show that both mRNA and protein levels of receptor-interacting protein 3 (RIP3) in the hearts are increased by I/R injury and doxorubicin (Dox) treatment. In mice, RIP3 deficiency ameliorates myocardial necroptosis and heart failure induced by I /R (30-min ischemia/4-h or 8-week reper-fusion) or Dox treatment (20 mg/kg or 5 mg/kg ×4, i.p.).RIP3 overexpression induces cardiomyocyte necroptosis evidenced by de-creased intracellular ATP level and increased lactate dehydrogenase concentration in cell culture medium .RIP3 triggers myocardial ne-croptosis via activation of Ca2+/calmodulin-dependent protein kinase II (CaMKII), rather than the well-established RIP3 partners, RIP1 and MLKL (mixed lineage kinase domain-like protein).Specifically, our data indicate that I/R and Dox markedly activate myo-cardial CaMKII in wild-type but not RIP3-deficient mice , and that CaMKII inhibition or RIP 3 deficiency protect the heart from I/R-and Dox-induced cardiomyocyte necroptosis , cardiac remodeling and heart failure .Mechanistically , RIP3 activates CaMKII via both di-rect phosphorylation and indirect reactive oxidative species-dependent oxidation , and subsequently triggers opening of the mitochondrial permeability transition pore ( mPTP) and myocardial necroptosis .CONCLUSION: These findings identify CaMKII as a novel RIP 3 substrate and delineate a RIP3-CaMKII-mPTP myocardial necroptosis pathway , a promising target for the treatment of cardiac ischemic and oxidative damage , and heart failure .

3-Bromopyruvate (3BP) is a new, promising anticancer alkylating agent with several notable functions. In addition to inhibiting key glycolysis enzymes including hexokinase II and lactate dehydrogenase (LDH), 3BP also selectively inhibits mitochondrial oxidative phosphorylation, angiogenesis, and energy production in cancer cells. Moreover, 3BP induces hydrogen peroxide generation in cancer cells (oxidative stress effect) and competes with the LDH substrates pyruvate and lactate. There is only one published human clinical study showing that 3BP was effective in treating fibrolamellar hepatocellular carcinoma. LDH is a good measure for tumor evaluation and predicts the outcome of treatment better than the presence of a residual tumor mass. According to the Warburg effect, LDH is responsible for lactate synthesis, which facilitates cancer cell survival, progression, aggressiveness, metastasis, and angiogenesis. Lactate produced through LDH activity fuels aerobic cell populations inside tumors via metabolic symbiosis. In melanoma, the most deadly skin cancer, 3BP induced necrotic celldeath in sensitive cels, whereas high glutathione (GSH) content made other melanoma cels resistant to 3BP. Concurrent use of a GSH depletor with 3BP killed resistant melanoma cells. Survival of melanoma patients was inversely associated with high serum LDH levels, which was reported to be highly predictive of melanoma treatment in randomized clinical trials. Here, we report a 28-year-old man presented with stage IV metastatic melanoma affecting the back, left pleura, and lung. The disease caused total destruction of the left lung and a high serum LDH level (4,283 U/L). After ethics committee approval and written patient consent, the patient received 3BP intravenous infusions (1-2.2 mg/kg), but the anticancer effect was minimal as indicated by a high serum LDH level. This may have been due to high tumor GSH content. On combining oral paracetamol, which depletes tumor GSH, with 3BP treatment, serum LDH level dropped maximally. Although a slow intravenous infusion of 3BP appeared to have minimal cytotoxicity, its anticancer efficacy via this delivery method was low. This was possibly due to high tumor GSH content, which was increased after concurrent use of the GSH depletor paracetamol. If the anticancer effectiveness of 3BP is less than expected, the combination with paracetamol may be needed to sensitize cancer cels to 3BP-induced effects.

绿原酸对Aβ引起的小脑颗粒细胞损伤的保护作用

目的：探讨绿原酸对淀粉样β蛋白(amyloid beta-protein，Aβ)引起的小脑颗粒细胞损伤的保护作用。方法：体外培养小脑颗粒细胞和腹腔巨噬细胞，将经过10μmol/L Aβ预处理48h的巨噬细胞上清液移入神经细胞中，同时加200μmol/L的绿原酸。培养2天后，采用MTT法、western bolt和乳酸脱氢酶检测试剂盒检测细胞活性、微管相关蛋白-2(microtubule-associated protein-2，MAP-2)表达水平和乳酸脱氢酶(lactic dehydrogenase，LDH)漏出量。结果：将Aβ预处理的巨噬细胞上清液移入到单独培养的小脑颗粒细胞48h后，可使小脑颗粒细胞活性明显降低，仅为正常的28.35%，加入绿原酸后，细胞活性明显增加达到正常的75.82%(P<0.01)。而在正常培养的神经细胞+Aβ组中，小脑颗粒细胞仅有少许死亡。在加入巨噬细胞上清液的小脑颗粒细胞组中，MAP-2表达量明显下降，仅为正常培养组的31.23%，加入绿原酸后，MAP-2的表达量则显著回升(P<0.01)，达到正常的79.77%。LDH漏出量明显增加，由402nkat/L增加到3046nkat/L，加入绿原酸后，可以使LDH漏出量明显下降，只有1285nkat/L。结论：①Aβ蛋白是通过激活巨噬细胞，使其释放毒性物质，从而引起神经细胞的损伤。②绿原酸对Aβ引起的神经细胞损伤具有保护作用。