A total of 189 male subjects with normal and abnormal semen quality were divided into seven groups according to WHO semen analysis criteria : Group I was composed of 62 subjects with normal semen parameters. They were subdivided into 4 sub-groups based on the sperm motility, such as subgroups I-1 with sperm motility between 41-50%, I -2 with motility between 51-60%, I -3 with motility 61-70%, and I-4 with motility over 71%. Group II was composed of 29 subjects with oligozoospermia of less than 20 X 106/ml of sperm density. Group III was composed of 9 subjects with asthenozoospermia of less than 40% of sperm motility. Group IV was composed of 13 subjects with teratozoospermia of more than 40% of abnormal morphology. Group V was composed of 31 subjects with testicular secretory azoospermia. Group VI was composed of 39 subjects with post-testicular obstructive azoospermia. Group VII was composed of 6 subjects with successful vasovasostomy and epididymovasostomy without pregnancy. Seminal plasma was separated from spermatozoa by centrifugation of fresh semen. The contents of protein and fructose and enzyme activities of hyaluronidase, a-amylase, b-glucuronidase, arylsulfatase, azocoll proteinase and acrosin were measured in each seminal plasma of the study groups. Enzyme activities and fructose content were compared based on their sperm motility and abnormalities of spermatozoa. The protein contents of seminal plasma of each group were nearly same. The fructose contents of both sub-group I-4 and group III were found to be very low compared with normal sperm group. Hyaluronidase originated from sperm acrosome and other accessory sex glands. It is related to increasing sperm motility by lowering seminal viscosity. The hyaluronidase activities of the sub-group I -4 and Group VII showed less than those of other groups. Alpha-amylase which cleaved long chain of carbohydrate was measured in each seminal plasma of the study groups. It did not affect lowering the viscosity of seminal plasma and sperm motility. Beta-glucuronidase originated from sperm acrosome and other accessory sex glands. It is related to increasing sperm motility by lowering seminal viscosity. The activity of b-glucuronidase was lowest in the sub-group I -4 and highest in the sub-group I-2 and Group III. Arylsulfatase activity was nearly same in all study groups. Azocoll proteinase activities, a non-specific neutral proteolytic enzyme, were negligible throughout the groups investigated in this study. This may indicate that lowering the viscosity of seminal plasma did not related to the proteolytic process. Acrosin was not contained in the seminal plasma. Disintegration of mucopolysaccharide is somewhat known to influence lowering. the viscosity in the seminal plasma. These findings suggest that sperm motility is affected by the enzyme activities and viscosity in the seminal plasma.
Acrosin ; Acrosome ; alpha-Amylases ; Asthenozoospermia ; Azoospermia ; Centrifugation ; Fructose ; Glucuronidase ; Humans ; Hyaluronoglucosaminidase ; Male ; Oligospermia ; Pregnancy ; Semen Analysis ; Semen* ; Sperm Motility* ; Spermatozoa* ; Vasovasostomy ; Viscosity

BACKGROUND: Gut microbiota is closely associated with development and exacerbation of inflammatory bowel diseases (IBD). The aim of this study was to investigate differences in gut microbiota depending on sex and changes of gut microbiota during IBD developments. METHODS: 16s rRNA metagenomic sequencing was performed for fecal materials from 8-week-old wild type (WT) and interleukin 10 (IL-10) knockout (KO) C57BL/6 mice of both sexes. Diversity indices, relative abundance of microbiota, and linear discriminant analysis effect size were examined to compare microbial communities between groups. Clustering of groups was performed by principal coordinates analysis (PCoA) and unweighted pair group method with arithmetic mean (UPGMA). Functional capabilities of microbiota were estimated using phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt) based on Kyoto Encyclopedia of Genes and Genomes database. RESULTS: PCoA and UPGMA tree analysis of beta-diversity demonstrated significant differences in gut microbiota between male and female groups of WT mice, but not of IL-10 KO mice. Firmicutes to Bacteroides ratio was higher in male group than that in female group in both WT mice and IL-10 KO mice. Phylum Proteobacteria significantly increased in female IL-10 KO mice than that in female WT mice. At species level, Lactobacillus murinus, Bacteroides acidifaciens, and Helicobacter hepaticus significantly increased in IL-10 KO mice than in WT mice. The relative abundance of beta-glucuronidase (K01195) was higher in female IL-10 KO mice than that in female WT mice by PICRUSt. CONCLUSIONS: Our results suggest that microbiota-host interactions might differ between sexes during development of IBD.
Animals ; Bacteroides ; Female ; Firmicutes ; Gastrointestinal Microbiome ; Genome ; Glucuronidase ; Helicobacter hepaticus ; Humans ; Inflammatory Bowel Diseases ; Interleukin-10 ; Lactobacillus ; Male ; Metagenomics ; Methods ; Mice ; Microbiota ; Proteobacteria ; Sequence Analysis ; Sex Characteristics ; Trees

The increasing demand of Chinese materia medica could not be supplied by wild resource, and the cultivated medicinal materials become popular, which led to decreased quality of many medicinal materials due to the difference of the circumstance between the wild and the cultivated. How to improve quality becomes key points of Chinese medicine resource. The leaves of Scutellaria baicalensis were sprayed with H₂O₂, the activities of superoxide dismutase (SOD) and catalase (CAT) changed little, but there had been a marked decrease of peroxidase (POD) and ascorbic oxidase (APX), which showed that the antioxidase system declined. Meanwhile, H₂O₂, as enhanced the expression of phenylalnine ammonialyase (PAL) and β-glucuronidase (GUS) as well as activity of PAL, promoted the biosynthesis and biotransformation of flavonoids. At the day 2 after treated, H₂O₂ of 0.004 μmol·L⁻¹ the contents of the baicalin and the wogonoside decreased slightly, but the contents of the baicalein and the wogonin increased significantly, the baicalein from 0.094% to 0.324%, the wogonin from 0.060% to 0.110%, i. e. increased 246% and 83.3%, respectively.
Ascorbate Oxidase ; metabolism ; Catalase ; metabolism ; Drugs, Chinese Herbal ; chemistry ; Flavanones ; analysis ; Flavonoids ; analysis ; Glucosides ; analysis ; Glucuronidase ; metabolism ; Hydrogen Peroxide ; Peroxidase ; metabolism ; Phenylalanine Ammonia-Lyase ; metabolism ; Scutellaria baicalensis ; metabolism ; Secondary Metabolism ; Superoxide Dismutase ; metabolism

β-Glucosidase activity assays constitute an important indicator for the early diagnosis of neonatal necrotizing enterocolitis and qualitative changes in medicinal plants. The drawbacks of the existing methods are high consumption of both time and reagents, complexity in operation, and requirement of expensive instruments and highly trained personnel. The present study provides a simplified, highly selective, and miniaturized glucometer-based strategy for the detection of β-glucosidase activity. Single-factor experiments showed that optimum β-glucosidase activity was exhibited at 50 °C and pH 5.0 in a citric acid-sodium citrate buffer when reacting with 0.03 g/mL salicin for 30 min. The procedure for detection was simplified without the need of a chromogenic reaction. Validation of the analytical method demonstrated that the accuracy, precision, repeatability, stability, and durability were good. The linear ranges of β-glucosidase in a buffer solution and rat serum were 0.0873-1.5498 U/mL and 0.4076-2.9019 U/mL, respectively. The proposed method was free from interference from β-dextranase, snailase, β-galactosidase, hemicellulase, and glucuronic acid released by baicalin. This demonstrated that the proposed assay had a higher selectivity than the conventional dinitrosalicylic acid (DNS) assay because of the specificity for salicin and unique recognition of glucose by a personal glucose meter. Miniaturization of the method resulted in a microassay for β-glucosidase activity. The easy-to-operate method was successfully used to detect a series of β-glucosidases extracted from bitter almonds and cultured by Aspergillus niger. In addition, the simplified and miniaturized glucometer-based assay has potential application in the point-of-care testing of β-glucosidase in many fields, including medical diagnostics, food safety, and environmental monitoring.
Animals ; Aspergillus niger ; Calibration ; Cellulase/analysis* ; Chemistry, Clinical/methods* ; Dextranase/analysis* ; Enterocolitis, Necrotizing/diagnosis* ; Equipment Design ; Flavonoids/analysis* ; Glucose/analysis* ; Glucuronic Acid/analysis* ; Glucuronidase/analysis* ; Glycoside Hydrolases/analysis* ; Hydrogen-Ion Concentration ; Linear Models ; Multienzyme Complexes/analysis* ; Plants, Medicinal ; Polygalacturonase/analysis* ; Rats ; Reproducibility of Results ; beta-Galactosidase/analysis* ; beta-Glucosidase/analysis*

OBJECTIVETo provide evidence for three-level prevention of cholelithiasis by means of observing the effects of some choleretics on bile compositions drained from patients with pigment gallstone.

METHODSTwenty-seven patients suffering from primary pigment gallstones and having received treatment of choledochostomies plus T-tube or endoscopic nasal bile drainage (ENBD) were divided equally into three groups, and administered respectively with Lidanling (the LDL group), ursodesoxycholic acid (the UDA group) and combination of LDL and UDA (the LDL + UDA group) through oral intake (7 patients in each group). Besides, 6 post-operational patients got no treatment with any drug were allocated in the control group. Bile of all the patients was collected before treatment and on the 1, 3, 5, 7 th day after the treatment started to detect levels of total bile acid (TBA), glycocholic acid (GCA), taurocholic acid (TCA), glycocholic cheno-desoxycholic acid (GCDCA), total bilirubin (TBIL), uncombined bilirubin (UCB), concentration of calcium ion (Ca(2+)) as well as the bacterio-genetic and endogenous beta-glucuronidase activity for comparing.

RESULTSLevels of TBA, GCA, TCA and GCDCA got gradually increased in the UDA group and the LDL + UDA group after treatment (P < 0.05), while those in the LDL group remained unchanged, showing an insignificant difference as compared with those in the control group. In the LDL group and the LDL + UDA group, TBIL gradually increased while UCB gradually decreased in the course of treatment (P < 0.05). Moreover, levels of Ca(2+) and endogenous beta-glucuronidase activity got significantly lowered (P < 0.05).

CONCLUSIONCombined use of LDL and UDA could elevate levels of TBA, GCA, TCA, GCDCA, enhance the excretion of TBIL in patients with pigment gallstone after bile drainage, lower levels of UCB and Ca(2+) and the activity of endogenous beta-glucuronidase in the bile, so as to reduce the possibility of stone formation of bile, and therefore, it could be used to prevent the production of pigment gallstone, especially to prevent post-operative recurrence of stones.

Adult ; Bile ; chemistry ; Bile Acids and Salts ; analysis ; Bilirubin ; analysis ; Calcium ; analysis ; Cholagogues and Choleretics ; pharmacology ; Choledochostomy ; Cysteic Acid ; analogs & derivatives ; pharmacology ; Drainage ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gallstones ; metabolism ; Glucuronidase ; analysis ; Glycocholic Acid ; analysis ; Humans ; Male ; Middle Aged ; Taurocholic Acid ; analysis ; Ursodeoxycholic Acid ; analogs & derivatives ; pharmacology

OBJECTIVETo detect the expressions of heparanase and nuclear factor kappa B p65 (NF-kappaB p65) in pancreatic adenocarcinomas and analyze their relation to patients' prognosis and the regulatory mechanism of NF-kappaB on heparanase expression.

METHODSHeparanase and NF-kappaB p65 proteins in the tumor and adjacent tissues were detected by immunohistochemistry in 48 patients with pancreatic adenocarcinoma and analyzed for their clinicopathological significance.

RESULTSHeparanase and NF-kappaB p65 proteins were found in 30 (62.5%) and 22 (45.9%) tumor specimens, respectively, a rate significantly higher than that in the adjacent tissues. High heparanase expression was closely related to advanced TNM stage (P=0.031), lymph node metastasis (P=0.003) and decreased 3-year postoperative survival (20.0% vs 0%, P=0.001). NF-kappaB p65 expression was associated with lymph node metastasis (P=0.017) and distant metastasis (P=0.031), but had a higher positive rate in heparanase-positive cases than in heparanase-negative cases (P=0.018). Multivariate analysis showed that neither heparanase nor NF-kappaB p65 was the independent prognostic factors.

CONCLUSIONHeparanase is overexpressed in pancreatic adenocarcinomas in association with decreased postoperative survival. NF-kappaB may up-regulate heparanase expression and promote heparanase-dependent tumor invasion and metastasis.

Adenocarcinoma ; diagnosis ; genetics ; Adult ; Aged ; Animals ; Female ; Gene Expression Regulation, Neoplastic ; Glucuronidase ; metabolism ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Pancreatic Neoplasms ; diagnosis ; genetics ; Prognosis ; Risk Factors ; Transcription Factor RelA ; metabolism

BACKGROUND/AIMS: The potential physiologic roles of Klotho in acute kidney injury (AKI) have recently been demonstrated in animal models. However, to date, there have been no human studies investigating the expression of renal Klotho in AKI. METHODS: We retrospectively collected biopsy specimens and clinical data of AKI patients between January 2001 and December 2012. Klotho expression was determined by immunohistochemical staining, and the clinical-pathological correlation was examined. RESULTS: Among the 34 patients diagnosed with acute tubular necrosis or acute tubulointerstitial nephritis, 21 patients without chronic histological lesions were included. The mean age was 37.3 +/- 18.5 years and the mean peak creatinine level was 8.2 +/- 5.5 mg/dL. In total, 10 patients (47.6%) received temporary renal replacement therapy (RRT); however, 17 patients (81%) showed functional recovery with creatinine levels of < 1.3 mg/dL after 1 month. The intensity of Klotho expression was scored as a percentage of Klotho-positive area. The renal Klotho score showed a significant negative correlation with the initial or peak creatinine level. When the patients were divided into three groups according to the Klotho score (low, middle, high), the low group had a significantly higher peak creatinine level and a more frequent requirement for RRT. However, the Klotho score was not a significant predictor of renal recovery. CONCLUSIONS: The results demonstrated that renal Klotho expression in humans decreased significantly according to the severity of AKI, regardless of the etiology, and that low expression was associated with a poor short-term outcome.
Acute Kidney Injury/diagnosis/etiology/*metabolism/physiopathology/therapy ; Adolescent ; Adult ; Biomarkers/analysis ; Biopsy ; Down-Regulation ; Female ; Glucuronidase/*analysis ; Humans ; Immunohistochemistry ; Kidney/*chemistry/pathology/physiopathology ; Kidney Tubular Necrosis, Acute/diagnosis/etiology/*metabolism/physiopathology/therapy ; Male ; Middle Aged ; Necrosis ; Predictive Value of Tests ; Recovery of Function ; Renal Replacement Therapy ; Retrospective Studies ; Risk Factors ; Severity of Illness Index ; Time Factors ; Treatment Outcome ; Young Adult

OBJECTIVETo study the developmental phase on the growth and active compounds in Scutellaria baicalensis.

METHODSeeds of wild plants were collected from Laiwu and sowed in Fangshan (Beijing) and Laiwu (Shandong). Samples of aerial and underground parts were collected in five growth periods of sprouts, seedlings, flowering, seed drop and withered periods respectively. The length of taproot, fresh weight of root, diameter of taproot and the length of stem were determined. The content of active compounds and total flavonoids were determined by HPLC and ultraviolet spectrophotometry respectively. The transcripted level of PAL1, PAL2, PAL3, C4H, 4CL, CHS, GUS and UBGAT were analyzed with RT-PCR.

RESULTThe results showed that the aerial part of S. baicalensis grew quickly before flowering stage, and the underground part grew mostly between the periods of flowering and withered. In the whole growing developmental periods, the content of total flavonoids was not changed significantly, the content of baicalin was increased gradually and the content of baicalein was decreased gradually. Expression level of PAL and 4CL was the highest in withered period, CHS was increased between flowering and seed drop and decreased in withered period.

CONCLUSIONSeedlings and withered periods may be the key phase affecting the growth and active compounds in S. baicalensis.

Acyltransferases ; genetics ; metabolism ; Chromatography, High Pressure Liquid ; Coenzyme A Ligases ; genetics ; metabolism ; Flavonoids ; analysis ; metabolism ; Flowers ; genetics ; growth & development ; metabolism ; Gene Expression Regulation, Developmental ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Plant ; Glucuronidase ; genetics ; metabolism ; Glucuronosyltransferase ; genetics ; metabolism ; Phenylalanine Ammonia-Lyase ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plant Roots ; genetics ; growth & development ; metabolism ; Plant Stems ; genetics ; growth & development ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Scutellaria baicalensis ; genetics ; growth & development ; metabolism ; Seedlings ; genetics ; growth & development ; metabolism ; Spectrophotometry, Ultraviolet ; Time Factors ; Trans-Cinnamate 4-Monooxygenase ; genetics ; metabolism