男性不育患者精浆miR-551b水平变化及其临床价值

目的 检测和分析男性不育症患者精浆miR-551b水平变化,探讨其作为男性不育症辅助分子标志物的可能性。方法 选取2008年11月～2015年3月在南京总医院生殖医学中心和江苏省中医院男科门诊确诊的92例男性不育症患者及同期招募的34例年龄匹配正常生育男性精液标本,记录患者及对照临床资料,测定患者及对照精液参数包括精子密度、精子活率、a级精子比例、a+b级精子比例及部分精浆生化指标α-糖苷、酸性磷酸酶、肉毒碱、果糖水平。实时荧光定量PCR技术(quantitative real-time PCR,qRT-PCR)检测和比较弱精症、非梗阻性无精症患者及正常生育男性精浆中miR-551b含量变化,统计分析精浆miR-551b临床价值及与精液参数、精浆生化指标相关性。结果 qRT-PCR结果显示,正常生育组精浆miR-551b水平为20.63(9.59,37.83)fmol/L,弱精症患者为62.29(25.22,101.43)fmol/L,较正常生育组显著升高(U=297.00); 无精症患者精浆miR-551b水平为4.70(2.41,13.71)fmol/L,较正常生育组明显降低(U=356.00),差异均具有统计学意义(P<0.001)。受试者工作特征曲线(receiver operating characteristic curve,ROC)分析显示,精浆miR-551b区分弱精症、无精症患者与正常生育组的ROC曲线下面积(AUC ROC)分别为0.810(95%CI 0.718～0.902)和0.772(95%CI 0.667～0.878); 鉴别弱精症和无精症患者的AUC ROC为0.932(95%CI 0.885～0.979)。Spearman秩相关性分析显示,男性不育患者精浆miR-551b水平与精子密度(r=0.735)、精子活率(r=0.643)、a级精子比例(r=0.672)、a+b级精子比例(r=0.682)及α-糖苷(r=0.375)呈正相关,差异均具有统计学意义(均P<0.05)。逐步多元线性回归结果显示,男性不育症患者精浆miR-551b水平与精子密度呈显著独立相关(β=0.618,P<0.001,校正r²=0.368)。逻辑回归分析显示,精浆miR-551b是弱精症[OR = 24.889(95%CI 5.302～116.843),P<0.001]和无精症[OR=6.303(95%CI 1.316～30.179),P=0.021]的潜在危险因素。结论 男性不育症患者精浆miR-551b水平与正常生育男性存在明显差异,且与精子密度和活力密切相关,有潜力成为男性不育患者辅助诊断和鉴别诊断的新型分子标志物。     

Thermodynamic study of beta-N-acetylhexosaminidase enzyme heterogeneity in human seminal plasma

BACKGROUND: It has been suggested that the activity of beta-N-acetylhexosaminidase (Hex) in seminal plasma may be used as a biochemical marker of azoospermia. The purpose of our study was to evaluate this hypothesis using a thermodynamic procedure developed to determine total Hex activity and that of its isoenzymes in this biological fluid. METHODS: Using the substrate 3,3'-dichlorophenolsulphoftaleinil N-acetyl-beta-D-glucosaminide, a highly significant difference (p<0.001) is found between the activation energy of Hex A (41.5 kJ/mol) and of Hex B (72.3 kJ/mol), making it possible to determine the activity of these isoenzymes from the apparent activation energy of the total Hex in seminal plasma. RESULTS: A significant difference between the normozoospermic and azoospermic groups was only found for Hex A isoenzyme activity (p<0.05), although with considerable overlapping between the values of both groups. Significant partial correlations were found for the total Hex, Hex A and Hex B activities with the immobile spermatozoa count (p<0.01) and for total Hex and Hex B with the dead spermatozoa count (p<0.05). In turn, Hex A had a significant partial correlation with the live spermatozoa count (p<0.05); however, Hex activity in seminal plasma of acromosomal origin appears to be of little importance in quantitative terms. CONCLUSIONS: It was not possible to confirm that total Hex activity in seminal plasma, or even of its isoenzymes Hex A and Hex B, is a suitable biochemical marker of azoospermia (efficiency< or =67%). The thermodynamic procedure described may be a useful alternative for the study of the Hex enzyme heterogeneity in spermatozoa.     

Relationship between lipocalin-type prostaglandin D synthase and alpha-glucosidase in azoospermia seminal plasma

BACKGROUND: [corrected] To determine the correlation of lipocalin-type prostaglandin D synthase (L-PGDS) and alpha-glucosidase in semen. METHODS: We analyzed 68 seminal plasmas for lipocalin-type prostaglandin D synthase (L-PGDS) and alpha-glucosidase, L-PGDS was analyzed by ELISA. The semen donors were categorized in 3 groups: normal, obstructive and non-obstructive azoospermia. We then evaluated their correlation. RESULTS: The difference of L-PGDS concentration (P<0.001) and alpha-glucosidase activity (P<0.001) among the 3 clinical groups was statistically significant. Correlation between L-PGDS concentration and alpha-glucosidase was also statistically significant. L-PGDS concentration correlated positively with alpha-glucosidase activity (r=0.882). CONCLUSIONS: L-PGDS in seminal plasma, like alpha-glucosidase, suggests an obstruction of the seminal ducts and may be a potential marker that may aid in the differential diagnosis of obstructive and non-obstructive azoospermia.     

Isoenzymes of urinary N-acetyl-beta-D-glucosaminidase (NAG) in patients with renal transplants

Renal transplant recipients were divided into five categories according to their clinical course from transplantation to their discharge from hospital. Total N-acetyl-beta-D-glucosaminidase (NAG) activity in urine was determined using a chromogenic substrate 2-methoxy-4-(2'-nitrovinyl)-phenyl 2-acetamido-2-deoxy-beta-D-glucopyranoside. The isoenzyme composition of the urine of each patient was determined by semi-automated DEAE-cellulose chromatography. Although raised NAG activity was found in stable transplant patients compared to controls, the level of activity was constant and no change in the isoenzyme profile was found. In reversible rejection there was a marked increase in the intermediate forms, particularly I2 and a concomitant fall in the relative amount of the A-form present but the profile became normal when the patient stabilised. Much more complex patterns were observed in patients who did not respond to treatment. Both the B and I forms were elevated with a fall in the A-form and in one case excretion of the serum As form was observed. The intermediate forms were always increased in rejection.     

Tay-Sachs disease: one-step assay of beta-N-acetylhexosaminidase in serum with a sulphated chromogenic substrate

A sulphated chromogenic compound, p-nitrophenyl-6-sulpho-2-acetamido-2-deoxy-beta-D-glucopyranoside, which can be hydrolysed enzymatically to p-nitrophenol and the sulphated amino sugar, was used as a substrate for the determination of activity of beta-N-acetylhexosaminidase isoenzymes in human serum. The sera of six Tay-Sachs patients lacking isoenzyme A and heat-inactivated control serum exhibited 6% of the mean normal enzyme activity of 1.32 U/l (1-s range = 1.07-1.57 U/l). In 10 obligate carriers of the Tay-Sachs gene the enzyme activity was 52% (1-s range = 45-60%) of the mean normal value. Therefore, by using the sulphated chromogenic substrate Tay-Sachs disease can be diagnosed enzymatically in a simple one-step procedure, but the 2-s activity ranges of heterozygotes and normals overlap. The assay is not absolutely specific for isoenzyme A of beta-N-acetylhexosaminidase, because the substrate can be hydrolysed to a certain extent by beta-N-acetylhexosaminidase I.     

存活素在正常及不育男性精浆中的检测及其意义

【目的】探讨健康生育男性及不育男性患者精液中存活素蛋白（一种凋亡抑制剂）的含量水平及临床意义。【方法】采用酶联免疫吸附试验检测23例健康生育男性志愿者、22例少弱精子症患者、39例非梗阻性无精症患者及12例梗阻性无精症患者精浆中的存活素含量。对梗阻性无精症患者和非梗阻性少精症患者进行睾丸活检,同时行组织病理学检查及睾丸穿刺抽吸术。【结果】精浆存活素水平在有生育能力的健康对照组中含量最高,其次为少弱精子症患者,而在非梗阻性无精症患者中含量少,差异有显著性（P〈0．05）。梗阻性无精症患者精浆存活素缺如,精浆中存活素的含量与精子发生及产量呈正相关,而与精液中异常形态精子的百分比呈负相关。在睾丸穿刺抽吸术成功的非梗阻性无精症患者精液中可检测到存活素,而在睾丸穿刺抽吸术不成功的患者中无法检出。【结论】精浆存活素是睾丸源性的,它与精子发生及活力有关。在非梗阻性无精症患者中,睾丸穿刺抽吸术检测是否存有精子与存活素相关。     

不育男性精浆中免疫球蛋白与精子参数的相关分析

目的 检测不育男性患者精浆中免疫球蛋白IgA、IgG、IgM和精子活力与密度,分析各参数之间的相关性,探讨其与男性不育的关系.方法 96例不育症患者分为弱精子症组、少精子症组和无精子症3组,以31例精液质量正常的已生育男性作为参照对象.利用免疫比浊法和精子质量图文分析系统分别检测精浆中IgA、IgG、IgM和精子活力与密度.结果 少精子症和无精子症患者精浆中IgM含量与正常组和弱精子症组比较显著减低(P＜0.01,P＜0.05);精浆中IgA、IgG和IgM之间两两相关(P＜0.01),IgM含量与精子密度显著相关(P＜0.05);不育症组与正常组比较,IgA、IgG和IgM相互之间的相关性减低.结论 精浆中三种免疫球蛋白含量的失衡及IgM含量减少可能是导致男性不育的原因之一,该结果可为男性不育的研究提供参考.     

Alcohol dehydrogenase (ADH) isoenzymes and aldehyde dehydrogenase (ALDH) activity in the sera of patients with liver cancer

The principal enzymes catalyzing the conversion of ethanol to acetate are alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). The activities of these enzymes are elevated in the serum during the course of alcoholism or cirrhosis. In previous investigations we have found elevated levels of ADH, ALDH, and class I ADH activity in liver cancer cells. It can suggest that these changes may be reflected by enzyme activity in the serum. In this work, the activity of ADH isoenzymes, and ALDH in the sera of patients with liver cancer was measured. Serum samples were taken from 64 patients (28 drinkers, 36 nondrinkers), with liver cancer. 25 patients had primary and 39 metastatic liver tumors. Total ADH activity was measured by photometric method with p-nitrosodimethylaniline (NDMA) as a substrate and ALDH activity by the fluorimetric method with 6-methoxy-2-naphtaldehyde as a substrate. For the measurement of the activity of class I and II isoenzymes we employed the fluorimetric methods, with class-specific fluorogenic substrates. The activity of class III ADH was measured by the photometric method with formaldehyde and class IV with m-nitrobenzaldehyde as a substrate. A statistically significant increase of class I ADH isoenzymes was found in the sera of cancer patients. The median activity of this class isoenzyme in the total cancer group increased about 51% (2.94 mU/L) in the comparison to the control level (1.43 mU/L). The activity of the class I ADH isoenzyme was significantly higher in the sera of patients with metastatic tumors than with primary cancers. The activity of this class in the sera of drinkers and group of moderate drinkers was significantly higher in comparison to the control group and higher in the sera of heavy drinkers when compared with moderate drinking patients. The total ADH activity was significantly higher (44%) among patients with cancer than healthy ones. The activity of class I ADH isoenzymes was elevated only in the serum of patients with metastatic liver cancer. This increase of activity seems to be caused by the enzyme released from liver cancer cells and primary tumors originating in other organs.     

The chitinase system from Trichomonas vaginalis as a potential target for antimicrobial therapy of urogenital trichomoniasis.

Chitinolytic activities in Trichomonas vaginalis membrane extracts were assessed by assays of three enzyme systems: N-acetyl-beta-D-hexosaminidase (NAHase), chitobiosidase and chitotriosidase. N-acetyl-beta-D-hexosaminidase was the enzyme that showed the highest specific activity. After successive subcutaneous inoculations into mice and parasite recovery in culture, the enzyme activities increased significantly with the number of inoculations for up to eight passages. In addition, enzyme activities were maximum at the logarithmic phase of growth. Glycol chitin, a chitinase substrate, enhanced all chitinolytic activities by about 30% and a clear-cut correlation is shown between the capacity for erythrocyte lysis by parasites and NAHase expression. Chitobiosidase and chitotriosidase activities were both inhibited at 58% and 100%, respectively, by allosamidine, a chitinase inhibitor used at 3 microM, whereas NAHase activity was not affected. Seven putative NAHase inhibitors (compounds n, 1-7), ureido and thioureido derivatives of 2-amino-2-deoxy-D-glucose were evaluated and five of them had K(i) values in the range 30-70 microM. The most active compound (compound 6) was functionally competitive with respect to the substrate with a K(i) value of 30 microM. The IC(50) values of the most active compounds on T. vaginalis were in the range 62-85 microM. These results indicate that chitinases of T. vaginalis are involved in pathogenicity and they could be an interesting target for drugs since chitinase inhibitors also inhibit parasite growth.     

男性不育症患者精浆中miR-145 的表达水平及其临床意义

目的　探讨男性不育症患者精浆中miR-145 的表达水平及其临床意义。方法　招募2017 年1 月~ 2018 年9 月 男性不育症患者130 例和同期体检正常的生育男性50 例为正常对照组。采用实时荧光定量PCR 检测精浆中miR-145 的 表达水平,精液常规参数采用WLJY-9000 伟力彩色精子检测系统进行分析。应用ROC 曲线分析miR-145 对男性不育症 的诊断价值,采用Pearson 相关分析精浆中miR-145 表达水平与精液参数的相关性。结果　男性不育症组精浆中miR- 145 表达水平明显高于对照组（3.92±0.86 vs 1.16±0.35）,而男性不育症组精子浓度（70.24±28.60 vs 112.50±46.35） ×106/mL、精子存活率（38.40±7.60 vs 73.84±5.82）%、前向运动精子百分率（17.52±9.73 vs 46.20±5.30）% 及正常 精子形态百分率（2.60±1.42 vs 9.25±1.70）% 明显低于对照组,差异均有统计学意义（t=4.852~11.238,均P<0.01）。 ROC 曲线分析显示,精浆中miR-145 表达水平诊断男性不育症的曲线下面积（AUC）为0.864（95%CI:0.807~0.925）, 其最佳诊断截断值为2.15,敏感度和特异度分别为92.8% 和75.0%。相关分析显示,男性不育症患者精浆中miR-145 表 达水平与精子浓度、精子存活率、前向运动精子百分率均呈正相关（r=0.427,0.604,0.538,均P<0.01）。结论　精浆 中miR-145 表达水平在男性不育症患者中明显上调,且精子浓度、精子存活率、前向运动精子百分率呈正相关,有望 作为男性不育症的辅助诊断指标。     

Plasma adenosine deaminase activity among HIV1 Clade C seropositives: relation to CD4 T cell population and antiretroviral therapy

BACKGROUND: Plasma adenosine deaminase and its isoenzymes(s) activities have been used as diagnostic marker for intracellular parasitism, including HIV infection, and malignancy of immune cells. HIV infection being primarily targeted against CD4 cells, it would be of interest to relate the activity of total plasma ADA and isoenzymes fractions to immune status and antiretroviral therapy. METHODS: In the present study, plasma total ADA activity (ADAT) including ADA1 and ADA2 isoenzyme(s) were assayed among HIV seropositive Clade C (n=90) comprising both asymptomatic (n=71) and symptomatic (n=19) and compared with that of HIV seronegatives (n=35). RESULTS: A significant increase in the activity of ADAT (16.30+/-0.80 v/s 6.18+/-0.30) as well as ADA1 (6.50+/-0.42 v/s 2.34+/-0.16) and ADA2 (9.79+/-0.53 v/s 3.85+/-0.23) isoenzyme(s) among the asymptomatic as well as the symptomatic subjects as compared to respective controls was noted. Increase in plasma ADAT activity, including ADA1 and ADA2 isoenzyme(s), were found to have negative correlation with CD4 counts (r, -0.273; p<0.05). The increased plasma ADAT activity among the asymptomatic HIV seropositive with CD4 counts>500 (13.2+/-1.65; p<0.01) as well as those who were on antiretroviral therapy (19.31+/-1.36; p<0.001) was evident. CONCLUSIONS: These findings suggest that plasma ADA can be a sensitive marker of an ongoing biological insult to host tissues either because of infection and/or side effects of medication. Measurement of plasma ADA activity, along with serological evidence for HIV infection may provide an alternate laboratory tool to monitor intracellular parasitism including secondary infection vis a vis the after effects of therapeutic outcome.     

Human seminal clusterin (SP-40,40). Isolation and characterization.

Molecular cloning of the human complement inhibitor SP-40,40, has revealed strong homology to a major rat and ram Sertoli cell product, sulfated glycoprotein-2, known also as clusterin. This study reports the purification and characterization of human seminal clusterin. Two-dimensional gel electrophoresis revealed charge differences between clusterin purified from semen and the serum-derived material. Both preparations demonstrate comparable hemagglutination (clustering) activity and inhibition of C5b-6 initiated hemolysis. The average clusterin concentration in normal seminal plasma is considerably higher than that found in serum. Mean seminal plasma clusterin concentrations were significantly lower in azoospermia caused by obstruction or seminiferous tubule failure than with oligospermia or normospermia. Only men with vasal agenesis had undetectable seminal clusterin, suggesting that some of the seminal clusterin is produced by the seminal vesicles. Immunofluorescence of human spermatozoa revealed that clusterin was detected on 10% of spermatozoa, predominantly those that were immature or had abnormal morphology. A pilot study of 25 patients suggests that seminal clusterin concentration, together with sperm motility and morphology, is correlated with the fertilization rate in vitro. The function of seminal clusterin is unknown. Its extensive distribution in the male genital tract and its high concentration in seminal plasma suggests an important role in male fertility.     

血浆肿瘤坏死因子受体相关因子-2升高与炎症性肠病内镜下疾病活动性的相关性分析

【目的】分析血浆肿瘤坏死因子受体相关因子-2（TRAF-2）水平对于炎症性肠病的诊断价值以及与内镜下疾病活动性的相关性。【方法】应用酶联免疫吸附试验（ELISA）分析炎症性肠病患者和正常对照者血浆中TRAF2蛋白的表达,受试者工作特征曲线（receiver-operating characteristic,ROc）分析血浆TRAF-2水平对于克罗恩病和溃疡性结肠炎的诊断价值,应用Pearson相关分析研究血浆TRAF-2水平与内镜下疾病活动性的相关性。数据处理使用GraphPad Prism5。【结果】克罗恩病患者和溃疡性结肠炎患者血浆TRAF-2水平显著高于正常对照者（P〈0．01）,同时TRAF-2对区分克罗恩病患者和正常对照者（P=0．001）以及区分溃疡性结肠炎患者和正常对照者（P=0．000）具有显著的诊断价值。克罗恩病患者（r=-0．097,P=0．455）以及溃疡性结肠炎患者（r=0．016,P=0．902）血浆TRA卜2表达水平和内镜下疾病活动指数均无显著的相关性。【结论】炎症性肠病患者血浆TRAF-2水平增高,对区分炎症性肠病患者和正常对照者具有诊断价值,但血浆中TRAF-2的水平不能反应内镜下疾病活动程度。     

精浆NAG和血清FSH联合检测在梗阻性无精子症诊治中的应用

目的：探讨精浆中性α糖苷酶（NAG）和血清促卵泡生成素（FSH）联合检测在梗阻性无精子症诊断中的意义。方法分析24例男性无精子症患者（14例为梗阻性无精子症,10例为非梗阻性无精子症）精浆NAG和FSH 水平,与16例健康已生育男性（正常对照组）的检测结果进行相关性分析,同时结合睾丸体积和详细的病史资料判断无精子症病因及分型。结果无精子症患者（包括梗阻性和非梗阻性无精子症组）与正常对照组年龄差异无统计学意义（P＞0．05）,非梗阻性无精子症组睾丸体积明显比梗阻性无精子症组和正常对照组小（P＜0．01）。无精子症患者NAG含量均明显低于正常对照组（P＜0．01）,FSH 水平均明显高于正常对照组（P＜0．01）;梗阻性无精子症组NAG和血清FSH 水平明显低于非梗阻性无精子症组（P＜0．01）。结论精浆NAG和血清FSH 联合检测具有经济、快速、安全、无创等优点,对无精子症的诊断和临床指导治疗具有重要意义。     

High-performance liquid chromatographic detection of lipid peroxidation in human seminal plasma and its application to male infertility

BACKGROUND: A simple and reliable high-performance liquid chromatographic (HPLC) method has been developed and validated for the analysis of malondialdehyde (MDA) in human seminal plasma. METHODS: After human seminal plasma is hydrolyzed, MDA is reacted with thiobarbituric acid (TBA) to form MDA(TBA)(2), a red-colored adduct with maximum absorbance at 532 nm. HPLC separation of the adduct in human seminal plasma was performed on a Lichrospher C(18) column. RESULTS: A mobile phase composed of 0.025 mol/l KH(2)PO(4) (pH 6.2)--methanol in the ratio 58:42 (v/v) was found to be the most suitable for this separation. Under the chromatographic conditions described, the MDA-TBA adduct had a retention time of approximately 4 min and good separation and detectability of MDA in human seminal plasma sample was obtained. The method proved to be linear calibration in the range of MDA from 0.10 to 2.50 micromol/l. The relative standard deviations of within- and between-assay for MDA analysis were 3.1% and 3.8%, respectively. The average recovery was 90.0-98.8% for the human seminal plasma samples. The method has been successfully applied to the study of the lipid peroxidation levels in the seminal plasma of male infertility. Semen samples were obtained from healthy volunteers and infertile males. Ejaculates were classified into studied subgroups and defined as: obstructive azoospermia, non-obstructive azoospermia, oligozoospermia, asthenozoospermia, oligoasthenozoospermia and oligoasthenoteratozoospermia. With the exception of obstructive azoospermic group, MDA concentrations of seminal plasma in control group had very significant difference with those in other infertile groups (P < 0.01). CONCLUSIONS: This indicated that lipid peroxidation could be harmful to male sperm and reproductive system, which may lead to male infertility.     

Multiple forms of alkaline phosphatase in plasma of hemodialysis patients.

We used quantitative assays to measure the activity of the bone, liver, and intestinal forms of alkaline phosphatase in plasma in 75 patients with endstage chronic renal failure undergoing hemodialysis. The results were correlated with radiological and other biochemical indices of bone disease and with biochemical indices of liver disease. The total activity of alkaline phosphatase in plasma increased in 28 patients. In 10 of these patients, nine of whom had increased activity of gamma-glutamyltransferase in plasma, the increase in total activity of alkaline phosphatase was from the liver isoenzyme alone (nine patients) or from the liver and bone isoenzymes together (one patient). Intestinal alkaline phosphatase in plasma, although greater than 23 U/L in eight patients, was solely responsible for the increase in total alkaline phosphatase in one patient (who had normal gamma-glutamyltransferase). Bone alkaline phosphatase in plasma was increased in 25 patients, seven of whom had normal total alkaline phosphatase, and was closely correlated (r = 0.78) with osteocalcin concentration in plasma, which was increased in a much greater proportion of patients (99%). Both total and bone alkaline phosphatase were correlated with parathyrin in plasma (r = 0.46 and 0.50, respectively) and with osteocalcin (r = 0.60 and 0.78, respectively). Osteocalcin and bone alkaline phosphatase, but not parathyrin, decreased with age, implying that the skeletal response to parathyrin may be age dependent. In patients with increased total alkaline phosphatase undergoing hemodialysis, the concurrent measurement of gamma-glutamyltransferase may help identify whether the enzyme increase originates from the liver or bone, but this approach wrongly identified the source of the increase in three of 28 patients. Therefore, we recommend a separate measurement of the bone isoenzyme of alkaline phosphatase.     

The association between plasma beta-hexosaminidase and its isoenzyme patterns and retinopathy in type 1 diabetes mellitus.

beta-Hexosaminidase and its isoenzyme patterns were investigated in plasma from patients with Type 1 diabetes mellitus. The patients were divided into three main groups matched for duration of diabetes: (a) proliferative retinopathy (b), progress of retinopathy within a two-year period (c) and with no background retinopathy. When all patients were compared to a reference group, a significant increase of plasma beta-hexosaminidase activity was found. Patients with proliferative retinopathy had significantly increased activity of plasma beta-hexosaminidase compared to the reference group but not compared to the other diabetic patients. The isoenzyme distribution was not different in any of the diabetic subgroups compared to the reference group. It was also shown that various degrees of diabetic nephropathy did not influence total plasma Hex or the isoenzyme pattern.     

beta-N-acetyl-D-glucosaminidase isoenzymes by chromatofocusing from serum and skin in diabetes

beta-N-Acetyl-D-glucosaminidase and its kinetic characteristics were determined in serum and skin from both diabetes mellitus and impaired glucose tolerance patients and controls. Mean total activity was reduced (p less than 0.05) in the skin of both patient groups. beta-N-Acetyl-D-glucosaminidase isoenzyme expression was investigated using chromatofocusing on PBE-94 coupled with automated enzyme assay. The isoenzyme profiles from serum showed two major forms (A and B) whose ratio varied from 2:1 in controls to 3:1 in diabetics. Moreover, diabetes mellitus and impaired glucose tolerance subjects displayed an intermediate (I) form. The A/B isoenzyme ratio was completely reversed in skin.     

Elevated lipid peroxidation and disturbed antioxidant enzyme activities in plasma and erythrocytes of patients with uterine cervicitis and myoma.

OBJECTIVES: We investigated whether oxidative stress is associated with human uterine cervicitis and uterine myoma. DESIGN AND METHODS: We measured lipid peroxidation and antioxidant enzymes in plasma and erythrocytes of cervicitis patients and myoma patients in comparison with matched controls. Thiobarbituric acid-reactive substances (TBARS), a measure of lipid peroxidation, were determined in plasma; glutathione peroxidase (GSHPx) and catalase in erythrocytes; and superoxide dismutase (SOD) in both plasma and erythrocytes. RESULTS: We showed that plasma TBARS were significantly higher (p < 0.05) in both cervicitis patients and myoma patients than in controls. Plasma TBARS were significantly (and negatively) correlated with plasma and erythrocyte T-SOD activities in cervicitis patients only. Plasma T-SOD activity was significantly lower in both groups of patients than in controls whereas erythrocyte T-SOD activity was only significantly lower in myoma patients. The lowered plasma T-SOD activity in the cervicitis patients was attributed to decreased Mn-SOD activity whereas the lowered plasma T-SOD activity in myoma patients was attributed to decreased activities of both Cu,Zn-SOD and Mn-SOD. Erythrocyte GSHPx activity was 14% higher (p < 0.05) in cervicitis patients and 11% lower (p > 0.05) in myoma patients than in controls; catalase activity was 10% higher (p > 0.05) in cervicitis patients and 13% lower (p > 0.05) in myoma patients than in controls. Neither erythrocyte GSHPx nor catalase activity was significantly correlated with plasma TBARS. CONCLUSIONS: The elevated lipid peroxidation and disturbed antioxidant enzyme activities demonstrate the potential of oxidative injury in patients with uterine cervicitis and myoma.     

Determination of cystyl-aminopeptidase. Isoenzymes in seminal plasma and serum of different origin.

The "oxytocinase" activity has been determined as the CAP (cystyl-aminopeptidase, EC 3.4.11.3) activity with several substrates. It was demonstrated that serum samples from pregnant women and patients with serious liver disease had increased CAP activities compared with normal serum. It was also shown that seminal plasma had very high CAP activity. The pH optimum for the enzymes from the various sources differed, as did the metal dependence. Furthermore, samples from patients with liver disease had higher CAP activities when assayed in buffers containing amino groups, in contrast to the other samples. Gel chromatography and gel electrophoresis revealed the presence of several isoenzymes. The serum from pregnant women contained an isoenzyme neither present in normal plasma nor in any of the other sources tested. It is possible that this isoenzymes represents the true oxytocinase. CAP and LAP (leucine aminopeptidase) activities were very well correlated in serum from pregnant women and seminal plasma, while the correlation for samples from patients with liver disease was not as good. When care was taken to determine the CAP activity at the right pH and with the appropriate buffer, the risk for interference from other enzymes was minimized in the determination of oxytocinase as CAP activity.