A PCR method was used to amplify the sequence encoding the mature peptide of?-mannanase of Bacillus subtilis. The gene was inserted into the Pichia pastoris vector pPIC9K, downstream of?-factor signal peptide sequence. The resultant recombinant plasmid pPIC9K-MAN was lineared by BglII digestion and introduced into the host Pichia pastoris GS115 by PEG method. After screen, the recombinant P. pastoris strain MAN22 was obtained and fermented in large scale 5L fermenter. The recombinant mannanase activity could reach to 1102IU/ml. The properties of the recombinant mannanase were characterized.

Objective The change of thyroid hormones affects male sexual function, but the underlying physiological mechanism is not yet clear.The aim of this study was to explore the association of thyroid hormones with abnormal penile neuro-electrophysiology in patients with premature ejaculation (PE).Methods This retrospective study included 52 acquired PE (APE) patients aged 35.36 ± 8.39 years and 24 healthy males aged 32.87±7.45 years.We measured the levels of thyroid hormones in all the subjects and, based on the latency time of penile sympathetic skin responses (PSSR), classified the APE patients into a normal (n=31) and an abnormal PSSR type (n=31).Results The concentration of total tetraiodothyronine (TT4) in the serum was significantly higher in the APE patients than in the normal control ([102.81±18.37] vs [92.68±11.56] nmol/L, P=0.018).The levels of serum TT4 and free tetraiodothyronine (FT4) were both markedly increased in the APE patients of the abnormal PSSR type as compared with those of the normal PSSR type ([113.28±20.89] vs [95.72±12.42] nmol/L, P<0.001 and [12.02±0.88] vs [10.81±1.63] nmol/L, P=0.003).The FT4 concentration was correlated negatively with the latency time of PSSR (r=-0.363, P=0.008) and positively with the Premature Ejaculation Diagnostic Tool (PEDT) scores (r=0.455, P=0.001).Conclusion Abnormal PSSR may be associated with elevated levels of thyroid hormones in APE patients, and regulation of thyroid hormone levels may help improve the symptoms of PE with abnormal PSSR.

Objective To investigate role of interleukin-1?(IL-1?),interleukin-8(IL-8),tumor necrosis factor-?((TNF-?)) and insulinlike growth factor-1(IGF-1) in meconium aspiration syndrome(MAS) in newborns.Methods The concentrations of(IL-1?),IL-8,TNF-? and IGF-1 in peripheral serum were detected in 32 newborns with MAS and 15 normal newborns by radio-immunity at the first,the third and seventh day after being born.Correlations between IGF-1 and changes of IL-1?,IL-8,TNF-? was analyzed.Results Compared with control group,the concentration of IL-1?,IL-8 and TNF-? increased significantly in newborns with MAS in three days after being born(all P0.05).On the other hand, the concentration of IGF-1 in newborns with MAS was obviously lower than that of normal newborns one week after birth.The concentration of IGF-1 showed negative correlations with that of IL-1?,IL-8 and TNF-?(r=-0.67,-0.61,-0.73 all P

Animals ; Animals, Newborn ; Cell Hypoxia ; Cell Lineage ; Cell Proliferation ; Cells, Cultured ; Oligodendroglia ; cytology ; Rats ; Rats, Sprague-Dawley

The efficacy and safety of uric-acid-lowering therapy (UALT) on slowing the progression of chronic kidney disease (CKD) accompanied by hyperuricemia were assessed. We searched Cochrane Library, PubMed, EMbase, CNKI, Wanfang and Vip databases up to November 15, 2012 for randomized controlled trials (RCTs) which compared the effect of UALT to control therapy in hyperuricemic patients secondary to CKD, and then performed quality evaluation and meta-analysis on the included studies. Seven RCTs involving 451 cases were included. UALT delayed the increase of serum creatinine (MD=-62.55 μmol/L, 95% CI: -98.10 to -26.99) and blood urea nitrogen (MD= -6.15 mmol/L, 95% CI: -8.17 to -4.13) as well as the decrease of glomerular filtration rate [MD=5.65 mL/(min·1.73 m2), 95% CI: 1.88 to 9.41], decreased systolic blood pressure (SBP) (MD= -6.08 mmHg, 95% CI: -11.67 to -0.49), and reduced the risk of the renal disease progression (RR=0.30, 95% CI: 0.19 to 0.46). However, there was no statistically significant difference in 24-h urinary protein quantity and diastolic blood pressure (P>0.05). We identified that UALT could delay the progression of CKD with secondary hyperuricemia. And this also indirectly proved that hyperuricemia was a risk factor for the CKD progression.

The identity of higher-order neurons and circuits playing an associative role to control renal function is notwell understood.We identified specific neural populations of rostral elements of brain regions that project multisynaptically to the kidneys in 3~ days after injecting a retrograde tracer pseudorabies virus (PRV)-614 into kidney of 13 adult male C57BL/6J strain mice.PRV-614 infected neurons were detected in a number of mesencephalic (e.g.central amygdala nucleus),telencephalic regions and motor cortex.These divisions included the preoptic area (POA),dorsomedial hypothalamus (DMH),lateral hypothalamus,arcuate nucleus (Arc),suprachiasmatic nucleus (SCN),periventricular hypothalamus (PeH),and rostral and caudal subdivision of the paraventricular nucleus of the hypothalamus (PVN).PRV-614/Tyrosine hydroxylase (TH) double-labeled cells were found within DMH,Arc,SCN,PeH,PVN,the anterodorsal and medial POA.A subset of neurons in PVN that participated in regulating sympathetic outflow to kidney was catecholaminergic or serotonergic.PRV-614 infected neurons within the PVN also contained arginine vasopressin or oxytocin.These data demonstrate the rostral elements of brain innervate the kidney by the neuroanatomical circuitry.

To investigate the effects of carbamazepine (CBZ) on the plasma concentrations of valproic acid (VPA) and its toxic metabolite 2-propyl-4-pentenoic acid (4-ene VPA) in epileptic patients, the plasma concentrations of VPA and 4-ene VPA were determined, and the effect of CBZ on pharmacokinetics of VPA was evaluated. All patients had been divided into two groups (VPA group, n = 87; and VPA+CBZ group, n = 19). As compared to VPA group, the combination of CBZ significantly (P < 0.01) decreased the trough concentration of VPA [VPA group, (69.5 +/- 28.8) microg x mL(-1); VPA+CBZ group, (46.3 +/- 25.6) microg x mL(-1)] and does-adjusted VPA trough concentration [VPA group, (4.89 +/- 2.21) microg x mL(-1) x mg(-1) x kg(-1); VPA+CBZ group, (3.14 +/- 1.74) microg x mL(-1) x mg(-1) x kg(-1)]. However, the addition of CBZ did not influence the concentration of 4-ene VPA. The present study revealed that coadministration of CBZ can reduce VPA plasma concentration and may impact VPA clinical effect, therefore therapeutic drug mornitoring of VPA should be used when combined use of CBZ and VPA.
Adolescent ; Adult ; Anticonvulsants ; blood ; pharmacokinetics ; therapeutic use ; Carbamazepine ; blood ; pharmacokinetics ; therapeutic use ; Drug Interactions ; Drug Therapy, Combination ; Epilepsy ; blood ; drug therapy ; Fatty Acids, Monounsaturated ; blood ; Female ; Humans ; Male ; Valproic Acid ; blood ; pharmacokinetics ; therapeutic use ; Young Adult

OBJECTIVE: To investigate the polymorphism of the TPH gene T3792A locus in Han ethnic group of north China and its application value in forensic science. METHODS: The polymorphism of T3792A locus of the TPH gene was analyzed by the ASPCR of blood samples from 173 unrelated individuals of north Chinese Han population. RESULTS: The distribution of the T3792A locus polymorphism of the TPH gene in Han ethnic group of north China followed the Hardy-Weinberg law, with the allele A and T gene frequency of 0.486 and 0.514, respectively. CONCLUSION The TPH gene T3792A locus shows a very good genetic polymorphism, and may be applied to individual identification and paternity testing.
Asian People/genetics* ; China/ethnology* ; Forensic Genetics ; Gene Frequency ; Humans ; Paternity ; Polymorphism, Genetic ; Tryptophan Hydroxylase/genetics*

OBJECTIVETo explore the correlation between single-nucleotide polymorphisms (SNPs) of telomerase reverse transcriptase (TERT) rs2736098 and rs2736100 and the susceptibility to hepatocellular carcinoma (HCC).

METHODSThis case-control study design included 1300 diagnosed HCC patients with HBsAg positive and 1344 HBsAg positive people as control-group.rs2736098 and rs2736100 on TERT were selected as research sites, whose polymorphisms were detected by TaqMan allelic discrimination assay. The OR values (95%CI) were calculated by logistic regression to compare the correlation between different genotype and susceptibility to HCC.

RESULTSThe distribution frequencies of three genotypes as GG, AG and AA on rs2736098 were separately 39.3% (500/1273), 44.2% (563/1273) and 16.5% (210/1273) in case group; while respectively 39.6% (526/1328), 45.5% (604/1328) and 14.9% (198/1328) in control group. The distribution frequencies of three genotypes as AA, AC and CC on rs2736100 were separately 33.7% (428/1269), 49.9% (633/1269) and 16.4% (208/1269) in case group; while respectively 34.0% (449/1322), 49.2% (651/1322) and 16.8% (222/1322) in control group. The multi-variates logistic regression analysis showed that there was no significant difference between rs2736098 mutated A carriers and genotype GG carriers in the susceptibility to HCC after adjusting by age, sex, smoking and drinking factors (rs2736098, AA + AG vs GG: adjusted OR = 1.00 (95%CI: 0.86 - 1.18)); and there was no significant different between rs2736100 mutated C carriers and genotype AA carriers in the susceptibility to HCC either (AC + CC vs AA: adjusted OR = 1.03 (95%CI: 0.87 - 1.22)).

CONCLUSIONThe polymorphisms of rs2736098 and rs2736100 on TERT may not play a landmark role in susceptibility to HCC among Chinese population.

Adult ; Asian Continental Ancestry Group ; genetics ; Carcinoma, Hepatocellular ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Liver Neoplasms ; genetics ; Logistic Models ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Risk Factors ; Telomerase ; genetics

AIMTo study the relationship of neurotoxicity of 6-hydroxydopamine (6-OHDA) and the function of glutamate transporter.

METHODSUsing in vivo microdialysis together with high performance liquid chromatography (HPLC) to detect the alteration of glutamate in the striatum and extracellular fluid of the PC12 cell. The rate of apoptosis and the activity of PC12 cells are read in a flow cytometer and a photometer for enzyme-labeled assays. The function of glutamate transporter is decided by detecting the ability of L-[3H]-glutamate uptake.

RESULTS6-OHDA was shown to induce apoptosis and decrease the activity of PC12 cells. Increased release of glutamate was also found in PC12 cells and the injured striatum of the PD rats. But glutamate uptake in PC12 cells and rat striatum synaptosomes are inhibited obviously.

CONCLUSIONThe neurotoxicity of 6-hydroxydopamine is associated with declined function of glutamate transporters, which may be one important pathogenesis mechanisms of Parkinson's disease.

Amino Acid Transport System X-AG ; drug effects ; Animals ; Apoptosis ; drug effects ; Corpus Striatum ; metabolism ; Glutamic Acid ; metabolism ; Male ; Oxidopamine ; toxicity ; PC12 Cells ; Parkinson Disease ; metabolism ; Rats ; Rats, Sprague-Dawley