Human seminal plasma is rich in potential biological markers for male infertility and male reproductive system diseases, which have an application value in the diagnosis and treatment of male infertility. The methods for the detection of semen biochemical markers have been developed from the manual, semi-automatic to the present automatic means. The automatic detection of semen biochemical markers is known for its advantages of simple reagent composition and small amount of reagents for each test, simple setting of parameters, whole automatic procedure with few errors, short detection time contributive to batch detection and reduction of manpower cost, simple calibration and quality control procedure to ensure accurate and reliable results, output of results in the order of the samples in favor of clinical diagnosis and treatment, and open reagents applicable to various automatic biochemistry analyzers. At present, the automatic method is applied in the detection of such semen biochemical markers as seminal plasma total and neutral alpha-glucosidase, acid phosphatase, fructose, γ-glutamyl transpeptidase, zinc, citric acid, uric acid, superoxide dismutase and carnitine, sperm acrosin and lactate dehydrogenase C4, and semen free elastase, which can be used to evaluate the secretory functions of the epididymis, seminal vesicle and prostate, sperm acrosome and energy metabolism function, seminal plasma antioxidative function, and infection or silent infection in the male genital tract.
Acid Phosphatase ; analysis ; Biomarkers ; analysis ; Carnitine ; analysis ; Citric Acid ; analysis ; Epididymis ; metabolism ; Fructose ; analysis ; Humans ; Infertility, Male ; diagnosis ; Isoenzymes ; L-Lactate Dehydrogenase ; Male ; Prostate ; metabolism ; Semen ; chemistry ; Seminal Vesicles ; Spermatozoa ; chemistry ; alpha-Glucosidases ; analysis ; gamma-Glutamyltransferase ; analysis

OBJECTIVETo assess the effects of traditional herbal formulae Sijunzi Decoction (, Sagunja-tang, SJZD), Siwu Decoction (, Samul-tang, SWD), Bawu Decoction (, Palmul-tang, BWD) and Shiquan Dabu Decoction (, Sipjeondaebo-tang, SDD) on the activities of human cytochrome P450 (CYP450), a drug-metabolizing enzyme.

METHODSHerbal formula water extracts were filtered and lyophilized after the powder extracts were dissolved in distilled water. The activities of major human CYP450 isozymes (CYP3A4, CYP2C19, CYP2D6 and CYP2E1) were measured using in vitro fluorescence-based enzyme assays. The inhibitory effects of the herbal formulas on the activities of CYP450 were characterized as half maximal inhibition concentration (IC) values.

RESULTSAll the tested herbal formulae inhibited CYP2C19 activity (IC: SJZD, 83.28 μg/mL; SWD, 235.54 μg/mL; BWD, 166.82 μg/mL; SDD, 178.19 μg/mL); SJZD (IC= 196.46 μg/mL), SWD (IC= 333.42 μg/mL) and SDD (IC= 163.42 μg/mL) inhibited CYP2E1-mediated metabolism; whereas BWD exhibited comparatively weak inhibition of CYP2E1 (IC= 501.78 μg/mL). None of the four herbal formulas significantly affected CYP3A4 or CYP2D6.

CONCLUSIONSThese results suggest that SJZD, SWD, BWD and SDD could potentially inhibit the metabolism of co-administered synthetic drugs whose primary route of elimination is via CYP2C19. In addition, clinically relevant pharmacokinetic interactions could occur when SJZD, SWD or SDD is co-administered with drugs metabolized by CYP2E1. Our findings provide information for the safety and effective clinical use of these four classic herbal formulas.

Cytochrome P-450 Enzyme System ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Hot Temperature ; Humans ; Inhibitory Concentration 50 ; Isoenzymes ; metabolism ; Plant Extracts ; pharmacology ; Water ; chemistry

We found that non-small-cell lung cancer (NSCLC) cells express high levels of multiple aldehyde dehydrogenase (ALDH) isoforms via an informatics analysis of metabolic enzymes in NSCLC and immunohistochemical staining of NSCLC clinical tumor samples. Using a multiple reaction-monitoring mass spectrometry analysis, we found that multiple ALDH isozymes were generally abundant in NSCLC cells compared with their levels in normal IMR-90 human lung cells. As a result of the catalytic reaction mediated by ALDH, NADH is produced as a by-product from the conversion of aldehyde to carboxylic acid. We hypothesized that the NADH produced by ALDH may be a reliable energy source for ATP production in NSCLC. This study revealed that NADH production by ALDH contributes significantly to ATP production in NSCLC. Furthermore, gossypol, a pan-ALDH inhibitor, markedly reduced the level of ATP. Gossypol combined with phenformin synergistically reduced the ATP levels, which efficiently induced cell death following cell cycle arrest.
Adenosine Triphosphate ; Aldehyde Dehydrogenase* ; Cell Cycle Checkpoints ; Cell Death ; Energy Metabolism* ; Gossypol ; Humans ; Informatics ; Isoenzymes ; Lung ; Lung Neoplasms ; Mass Spectrometry ; NAD ; Phenformin ; Protein Isoforms

OBJECTIVETo investigate the expression of ALDH1, CXCR4 and E-cadherin in gastric carcinoma and their roles in lymphatic metastasis.

METHODSSurgical specimens from 127 cases of gastric carcinoma were examined for expressions of ALDH1, CXCR4 and E-cadherin immuohistochemistry with 60 adjacent tissues as control. The associations of ALDH1, CXCR4 and E-cadherin with the clinicopathological pfeatures, 5-year survival rate and lymph node metastasis of the patients were analyzed.

RESULTSALDH1, CXCR4 and E-cadherin were positive in 57.5% (73/127), 63.8% (81/127), and 36.2% (46/127) of the gastric carcinoma tissues, respectively, showing significant differences from the rates in the adjacent tissues (P<0.05). The expression of ALDH1 was significantly correlated with TNM stage and lymph node metastasis (P<0.05), CXCR4 was significantly correlated with the invasion depth, differentiation, TNM stage and lymph node metastasis of the tumor (P<0.05), and E-cadherin was significantly correlated with the invasion depth, differentiation and lymph node metastasis (P<0.05). The positivity rates of ALDH1, CXCR4 and E-cadherin were higher in cases with lymph node metastasis than in those without metastasis. E-cadherin expression was inversely correlated with ALDH1 and CXCR4 expression, and the latter two were positively correlated (P<0.001). Overexpressions of ALDH1 and CXCR4 and a decreased expression of E-cadherin were all related to a poor prognosis of the patients (P<0.05). The expressions ofALDH1, CXCR4 and E-cadherin were all independent prognostic factors of gastric carcinoma.

CONCLUSIONThe expressions of ALDH1, CXCR4 and E-cadherin are associated with the invasion, metastasis and prognosis of gastric carcinoma, and their combined detection provides important evidence for predicting the progression and prognosis of gastric carcinoma.

Cadherins ; genetics ; metabolism ; Carcinoma ; genetics ; metabolism ; Disease Progression ; Humans ; Isoenzymes ; genetics ; metabolism ; Lymphatic Metastasis ; Prognosis ; Receptors, CXCR4 ; genetics ; metabolism ; Retinal Dehydrogenase ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism ; Survival Rate

RNA binding proteins (RBPs) regulate the function of cells by interacting with nascent transcripts and therefore are receiving increasing attention from researchers for their roles in tissue development and homeostasis. The polypyrimidine tract binding (PTB) protein family of RBPs are important posttranscriptional regulators of gene expression. Further investigations on the post-transcriptional regulation mechanisms and isoforms of PTB proteins in the spermatogenesis show that PTB protein 1 (Ptbp1) is a predominant isoform in mitotic cells (spermatogonia), while Ptbp2 predominates in meiotic spermatocytes and postmeiotic spermatids and binds to the specific 3' untranslated region (3' UTR) of the phosphoglycerate kinase 2 (Pgk-2) mRNA, which helps to stabilize Pgk-2 mRNA in male mouse germ cells. In case of Ptbp2 inactivation in the testis, the differentiation of germ cells arrests in the stage of round spermatids, with proliferation of multinucleated cells in the seminiferous tubule, increased apoptosis of spermatocytes, atrophy of seminiferous tubules, and lack of elongating spermatids, which consequently affects male fertility. This article presents an overview on the structure of the PTB protein and its role in regulating mammalian spermatogenesis.
Animals ; Atrophy ; Gene Expression Regulation ; physiology ; Heterogeneous-Nuclear Ribonucleoproteins ; metabolism ; physiology ; Homeostasis ; Isoenzymes ; metabolism ; Male ; Mice ; Nerve Tissue Proteins ; metabolism ; physiology ; Phosphoglycerate Kinase ; metabolism ; Polypyrimidine Tract-Binding Protein ; metabolism ; physiology ; RNA, Messenger ; metabolism ; RNA-Binding Proteins ; Seminiferous Tubules ; pathology ; Spermatids ; metabolism ; Spermatocytes ; metabolism ; Spermatogenesis ; physiology ; Spermatogonia ; metabolism ; Testis ; metabolism

To determine expression of lactate dehydrogenase (LDH)-5 in non-Hodgkin lymphoma and its clinical significance.
 Methods: LDH-5 levels and LDH levels in NHL patients were examined by agarose gel electrophoresis and enzymatic method (n=63), respectively. Positive rates of LDH-5 and LDH were statistically analyzed.
 Results: The median age of NHL patients was 56(19-84) years old, including 36 males and 27 females. The positive numbers for LDH-5 and LDH in the initial treatment group (n=43) were significantly different (P<0.05). There was significant difference in 22 cases of diffuse large B cell lymphoma and in 9 cases of T cell lymphoma, whereas there was not significant difference in 12 cases of small B cell lymphoma (P>0.05). In 15 cases under the status of progress, the difference of LDH-5 and LDH expressions were not significant (P>0.05), whereas the difference in cases of small B cell lymphoma was significant (P<0.05).
 Conclusion: LDH-5 can be used as an index for NHL to judge the tumor load and to predict the recurrence.
Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; blood ; metabolism ; Female ; Humans ; Isoenzymes ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Lactate Dehydrogenase 5 ; Lymphoma, B-Cell ; genetics ; Lymphoma, Large B-Cell, Diffuse ; genetics ; Lymphoma, Non-Hodgkin ; genetics ; Lymphoma, T-Cell ; genetics ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; genetics ; Prognosis ; Tumor Burden

To research the influence of Reduning injection on the activity and mRNA expression of cytochrome P450 (CYP450) system in rat liver microsomes. Rat liver microsomes were prepared after a seven-days continuous administration of Reduning injection. An HPLC-MS method was applied to determine the specific metabolites of CYP450 probe substrates in rat liver microsomal incubations. The activity of CYP450 isozymes were represented by the formation of metabolites. The Real-time quantitative polymerase chain reaction (Q-PCR) was applied to determine the mRNA expression levels of CYP450. Reduning injection significantly reduced the activity of CYP2B1, 2C12, 2C13 (P < 0.01), but did not affect CYPlA2; low dose and high dose of Reduning injection had an inhibition trend on the activity of CYP2D2, but did not statistically differ from control group; low dose of Reduning injection significantly induced the activity of CYP3A1 (P < 0.01), high dose of Reduning injection had an induce trend on the activity of CYP3A1, but did not statistically differ from control. At the mRNA level, low and high dose of Reduning injection had an induce trend on the expression of CYP1A2, 2C11, 2D1, 2E1, 3A1, but did not statistically differ from control. Reduning injection significantly induced the activity of CYP2B1. Reduning injection significantly induced the activity of CYP3A1 in mRNA expression and enzyme activity levels, which may result adverse drug reaction after being combined with macrolides antibiotics. Reduning injection significantly reduced the activity of CYP2B1, 2C12, 2C13, 2D2 in enzyme activity levels, when combined with other drugs, it should be fully taken into account of the possible drug-drug interaction in order to avoid adverse side effects.
Animals ; Cytochrome P-450 Enzyme System ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Injections ; Isoenzymes ; metabolism ; Male ; Microsomes, Liver ; drug effects ; enzymology ; Rats ; Rats, Sprague-Dawley

Osteoclast-like cells are known to inhibit arterial calcification. Receptor activator of NF-κB ligand (RANKL) is likely to act as an inducer of osteoclast-like cell differentiation. However, several studies have shown that RANKL promotes arterial calcification rather than inhibiting arterial calcification. The present study was conducted in order to investigate and elucidate this paradox. Firstly, RANKL was added into the media, and the monocyte precursor cells were cultured. Morphological observation and Tartrate resistant acid phosphatase (TRAP) staining were used to assess whether RANKL could induce the monocyte precursor cells to differentiate into osteoclast-like cells. During arterial calcification, in vivo and in vitro expression of RANKL and its inhibitor, osteoprotegerin (OPG), was detected by real-time PCR. The extent of osteoclast-like cell differentiation was also assessed. It was found RANKL could induce osteoclast-like cell differentiation. There was no in vivo or in vitro expression of osteoclast-like cells in the early stage of calcification. At that time, the ratio of RANKL to OPG was very low. In the late stage of calcification, a small amount of osteoclast-like cell expression coincided with a relatively high ratio of RANKL to OPG. According to the results, the ratio of RANKL to OPG was very low during most of the arterial calcification period. This made it possible for OPG to completely inhibit RANKL-induced osteoclast-like cell differentiation. This likely explains why RANKL had the ability to induce osteoclast-like cell differentiation but acted as a promoter of calcification instead.
Acid Phosphatase ; genetics ; metabolism ; Animals ; Aorta ; drug effects ; metabolism ; pathology ; Cell Differentiation ; Coculture Techniques ; Gene Expression Regulation ; Isoenzymes ; genetics ; metabolism ; Male ; Monocytes ; cytology ; drug effects ; metabolism ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; pathology ; Osteoclasts ; drug effects ; metabolism ; pathology ; Osteoprotegerin ; genetics ; metabolism ; RANK Ligand ; genetics ; metabolism ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Tartrate-Resistant Acid Phosphatase ; Vascular Calcification ; genetics ; metabolism ; pathology

Impaired glucose homeostasis is one of the risk factors for causing metabolic diseases including obesity, type 2 diabetes, and cancers. In glucose metabolism, pyruvate dehydrogenase complex (PDC) mediates a major regulatory step, an irreversible reaction of oxidative decarboxylation of pyruvate to acetyl-CoA. Tight control of PDC is critical because it plays a key role in glucose disposal. PDC activity is tightly regulated using phosphorylation by pyruvate dehydrogenase kinases (PDK1 to 4) and pyruvate dehydrogenase phosphatases (PDP1 and 2). PDKs and PDPs exhibit unique tissue expression patterns, kinetic properties, and sensitivities to regulatory molecules. During the last decades, the up-regulation of PDKs has been observed in the tissues of patients and mammals with metabolic diseases, which suggests that the inhibition of these kinases may have beneficial effects for treating metabolic diseases. This review summarizes the recent advances in the role of specific PDK isoenzymes on the induction of metabolic diseases and describes the effects of PDK inhibition on the prevention of metabolic diseases using pharmacological inhibitors. Based on these reports, PDK isoenzymes are strong therapeutic targets for preventing and treating metabolic diseases.
Acetyl Coenzyme A ; Decarboxylation ; Diabetes Mellitus, Type 2 ; Glucose ; Homeostasis ; Humans ; Isoenzymes ; Mammals ; Metabolic Diseases ; Metabolism ; Obesity ; Oxidoreductases* ; Phosphoric Monoester Hydrolases ; Phosphorylation ; Phosphotransferases* ; Pyruvate Dehydrogenase Complex ; Pyruvic Acid* ; Risk Factors ; Up-Regulation

BACKGROUNDIt has been demonstrated that only 10%-20% cigarette smokers finally suffer chronic obstructive pulmonary disease (COPD). The underlying mechanism of development remains uncertain so far. Nitric oxide (NO) has been found to be closely associated with the pathogenesis of COPD, the alteration of NO synthase (NOS) expression need to be revealed. The study aimed to investigate the alterations of NOS isoforms expressions between smokers with and without COPD, which might be helpful for identifying the susceptibility of smokers developing into COPD.

METHODSPeripheral lung tissues were obtained from 10 nonsmoker control subjects, 15 non-COPD smokers, and 15 smokers with COPD. Neuronal NOS (nNOS), inducible NOS (iNOS), and endothelial NOS (eNOS) mRNA and protein levels were measured in each sample by using real-time polymerase chain reaction and Western blotting.

RESULTSINOS mRNA was significantly increased in patients with COPD compared with nonsmokers and smokers with normal lung function (P < 0.001, P = 0.001, respectively). iNOS protein was also higher in COPD patients than nonsmokers and smokers with normal lung function (P < 0.01 and P = 0.01, respectively). However, expressions of nNOS and eNOS did not differ among nonsmokers, smokers with and without COPD. Furthermore, there was a negative correlation between iNOS protein level and lung function parameters forced expiratory volume in 1 s (FEV₁) (% predicted) (r = -0.549, P = 0.001) and FEV₁/forced vital capacity (%, r = -0.535, P = 0.001).

CONCLUSIONSThe expression of iNOS significantly increased in smokers with COPD compared with that in nonsmokers or smokers without COPD. The results suggest that iNOS might be involved in the pathogenesis of COPD, and may be a potential marker to identify the smokers who have more liability to suffer COPD.

Adult ; Aged ; Blotting, Western ; Female ; Humans ; Isoenzymes ; genetics ; metabolism ; Lung ; enzymology ; pathology ; Male ; Middle Aged ; Nitric Oxide Synthase ; genetics ; metabolism ; Pulmonary Disease, Chronic Obstructive ; enzymology ; pathology ; Real-Time Polymerase Chain Reaction