Objective To investigate the clinical value of observing the differences of GSK‐3βprotein expression and activity lev‐el in the peripheral blood mononuclear cells between type 2 diabetes mellitus patients and healthy people .Methods Using ELISA to detect the GSK‐3βprotein expression and activity levels of people who contained 30 patients with type 2 diabetes whose blood glu‐cose were not controlled (test group 1) ,30 cases of patients with type 2 diabetes whose blood glucose were controlled (test group 2) and 30 cases of healthy human and all the data were analyzed .Results The GSK‐3βprotein content of Type 2 diabetes mellitus group was higher than the control group(P<0 .05) ,the activity level of GSK‐3βin test group 1 was higher than the control group and test group(P<0 .05) .The GSK‐3β protein content of test group 2 was higher than the control group(P< 0 .05) .The male GSK‐3βactivity level in the test group 1 was higher than the men′s in the control group(P<0 .05) .Conclusion The GSK‐3βprotein content has a high expression in the type 2 diabetes mellitus patients .After drug treatment ,the patients ,who have controlled the blood glu‐cose effectively ,still showes high expression of the GSK‐3βprotein content ,but the activity levels showes a significant decline .In male pa‐tients with type 2 diabetes blood sugar in the control group of GSK‐3βshowes significantly higher activity level .

OBJECTIVETo determine the content of linarin in Yuye Jiedu granule.

METHODA HPLC method was developed. The chromatographic conditions are as follows Luna C18 column and acetonitrile-0.05 mol x L(-1) phosphate buffer-phosphoric acid (30:70:0.06) as mobil phase, detection wavelenth at 327 nm.

RESULTThe linear range of linarin was 0.025-0.50 microg. The average recovery was 98.7% and RSD 2.7%.

CONCLUSIONThe method is simple and accurate, with good repeatability, and can be used for determination of linarin in Yuye Jiedu granule.

Chromatography, High Pressure Liquid ; methods ; Chrysanthemum ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Glycosides ; analysis ; Lonicera ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control

Purpose To investigate the serum prostate specific antigen( PSA)feature of high grade prostatic intraepithe1ia1 neop1asia ( HGPIN)patients,and the association of the number of cores positive for HGPIN on initia1 biopsy and the risk of cancer deve1opment in second biopsy. Methods 492 cases of patients with suspicious prostate cancer were schedu1ed for transrecta1 u1trasound prostatic biopsy with an 8-core temp1ate. In the first biopsy,186 cases of patients with PCa,34 cases of patients with iso1ated HGPIN( on1y one core invo1ved with HGPIN)and 13 cases of patients with extensive HGPIN( two or more cores invo1ved with HGPIN),64 cases of pa-tients with LGPIN,195 cases of patients with BPH. The va1ues of PSA were ana1yzed and compared within these groups. In patients with extensive HGPIN or iso1ated HGPIN we proposed a repeat 8-core biopsy after 6 months independent of serum PSA 1eve1. The same measure was app1ied for patients diagnosed as LGPIN or BPH in the first biopsy with accompanying increase or persistent e1evation of serum PSA 1eve1. The incidence of PCa was ana1yzed and compared within these groups. Results The serum PSA 1eve1s were no sig-nificant1y different between LGPIN and BPH(P>0. 05),between iso1ated HGPIN and LGPIN(P>0. 05),and between iso1ated HG-PIN and BPH(P>0. 05). The serum PSA 1eve1s were significant1y different between extensive HGPIN and LGPIN(P<0. 05),be-tween extensive HGPIN and BPH(P<0. 05),and between extensive HGPIN and iso1ated HGPIN(P<0. 05). In the second biopsy, the incidence of PCa in patients with extensive HGPIN was 38. 48%,that in patients with iso1ated HGPIN was 9. 68%,that in patients with LGPIN was 12. 50%,and that in patients with BPH was 12. 20%. Conclusions The features of PSA in patients with iso1ated HGPIN are simi1ar to BPH,PSA 1eve1 in patients with extensive HGPIN were between PCa and BPH,and patients with extensive HG-PIN have a higher incidence of PCa in second biopsy than iso1ated HGPIN and BPH.

The expression of Attractin mRNA and protein in testis and semen of human and male mice was investigated. Human testis and semen samples were all collected from Reproductive Center of Renmin Hospital, Wuhan University in December, 2012. Testis samples were collected from 7 cases of obstructive azoospermias when they were subjected to diagnosed testis biopsy, and 30 normal human semen samples were obtained from those cases of semen analysis. Adult mice testis tissues were obtained from 10 2-month-old male BALB/c mice, and 60 male mice at different ages were classified into 10 groups (day 1, 5, 10, 15, 21, 28, 35, 42, 56, and 120 respectively, n=6 each). The expression of Attractin mRNA and protein in testis was detected by RT-PCR and Western blotting respectively. Human semen samples were centrifuged into sperm plasma (SP) and sperm extract (SE), and mice sperm samples were collected from the epididymis of 10 adult male BALB/c mice. Western blotting was used to determine the Attractin protein expression level. Attractin mRNA and protein were expressed in the testis of both patients with obstructive azoospermias and adult Bcl/B mice. Quantitative RT-PCR revealed that no Attractin mRNA was detectable in day 1 male BALB/c mice group. The Attractin mRNA and protein levels were low on the day 10, and increased with age until day 56. On the day 120, the expression levels of Attractin were decreased. As for human semen samples, Attractin protein was expressed in both SP and SE, but didn't exist in samples from the epididymis of male BALB/c mice. It was suggested that Attractin acted as a novel active substance and was involved in male reproduction in both human and BALB/c mice, but it exerted a different expression profile in different mammal species.

A high activity isoflavone-glucosidase, which hydrolysis glycosides, was obtainde using liquid fermentation from Absidia sp. R strain. The isoflavone-glucosidase was purified 11 folds with yielding rate of 10.9% after ammonium sulfate precipitation and DEAE-Cellocuse (DE-52) ion exchange chromatography. SDS-PAGE results showed that the molecular weight is 53kD. And the optimum temperature, the optimum pH, Km and pI of the enzyme are 50 deegrees C, 5.0, 1.3 x 10(-2) mol/L and 3.2, respectively. The isoflavone-glucosidase is also rather stable under 60 degrees C and in pH range from 5.0 to 7.0. The enzyme can be activated by Co2+ and Ca2+, and be inhibited by Ag+ and Cu2+.
Absidia ; enzymology ; Glucosidases ; isolation & purification ; metabolism ; Hydrogen-Ion Concentration ; Isoflavones ; metabolism ; Temperature

In order to study the effects of phosphorothioated antisense oligodeoxynucleotides (ASODN) on the expression of VEGF in human lymphoma cell line Namalwa cells, human lymphoma cell line Namalwa cells were incubated with VEGF ASODN (the final concentrations of VEGF ASODN were 5, 10, 20 micromol/L respectively), or scrambled sequence for 24 or 48 hours. The expressions of VEGF mRNA and VEGF protein were detected by reverse transcriptase-polymerase chain reaction and streptavidin peroxidase (SP) immunohistochemistry respectively. The results showed that the expression levels of VEGF mRNA in Namalwa cells treated with three concentration levels (5, 10, 20 micromol/L of ASODN) were 1.38, 0.96 and 0.57 respectively. Those in PBS-treated cells and scrambled sequence treated cells were 1.79 and 1.84. When treated with 20 micromol/L VEGF ASODN for 48 hours, VEGF protein of Namalwa cells decreased greatly. Meanwhile, there was no obvious change in the scrambled sequence treated group. It is concluded that VEGF ASODN can suppress the VEGF expression in Namalwa cells in vitro.
Cell Line, Tumor ; Humans ; Lymphoma ; metabolism ; pathology ; Oligonucleotides, Antisense ; pharmacology ; RNA, Messenger ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; metabolism

AIMTo study the relationship between resonance Rayleigh scattering(RRS) and the formation of association nanoparticle, and to develop a new and sensitive RRS method for the determination of trace berberine (BB).

METHODSThe association nanoparticle between BB and tetraphenylboron (TPB) was investigated by means of RRS, absorption spectral method and transmission electron microscope (TEM).

RESULTSIn pH 5.0 NaAc-HAc buffer solution, BB and TPB combine to BB-TPB association complex. The association complexes aggregate to (BB-TPB)n association nanoparticles, due to the strong hydrophobic force and Van der Waals force. It exhibits a resonance Rayleigh scattering peak at 470 nm, a maximum absorption peak at 368 nm. A new RRS method was proposed for the determination of 0.06 to 5.28 mg.L-1 BB in real samples with satisfactory results. The detection limit is 26 micrograms.L-1.

CONCLUSIONThe TEM of (BBjAgp-TPBj + p)h composite association nanoparticles was observed by means of composite association nanoreaction both TPB- and BB+ or Ag+. The results indicated that the formation of (BB-TPB)n association nanoparticles and interface of solid-liquid results in the enhanced resonance light-scattering. This new RRS method showed high sensitivity, and had been successfully applied to the determination of BB in real samples.

Berberine ; administration & dosage ; analysis ; Drug Carriers ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; Nanotechnology ; Scattering, Radiation ; Spectrum Analysis ; Tablets ; Technology, Pharmaceutical ; methods ; Tetraphenylborate ; chemistry

OBJECTIVETo study the volatile regularity of menthol and borneol in granule and lozenge at different temperature.

METHODKinetic method with GC being detection technique.

RESULTThe volatility of menthol and borneol acts as a pseudo first-order reaction in open system. Under the same condition, the volatile rate of menthol and borneol in granule is four times as fast as that in lozenge, and the volatile rate of borneol is faster than that of menthol.

CONCLUSIONThis study can be applied to improve the quality of lozenges containing menthol or/and borneol.

Bornanes ; administration & dosage ; chemistry ; Cyclodextrins ; Drug Carriers ; Drug Stability ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Menthol ; administration & dosage ; chemistry ; Tablets ; Temperature ; Volatilization ; beta-Cyclodextrins

A variety of natural and artificial cryoprotectant extenders have been explored to enhance sperm recovery following cryopreservation-thawing process.The current investigation is aimed at evaluating the effect of acetyl-L-carnitine on human spermatozoa and reactive species oxygen (ROS) level after freezing-thawing process.The spermatozoa were collected from 35 male patients diagnosed as having asthenospermia.The cryopreservation of human spermatozoa treated with acetyl-L-camitine at different concentrations (group B:2.5 mmol/L,group C:7.5 mmol/L,group D:15 mmol/L) was compared with control (group A:no acetyl-L-carnitine given).For the frozen-thawed spermatozoa,the viability,motility and DNA integrity were measured by comet assay,acrosome integrity by FITC-PNA staining and ROS level was determined in each group.The results showed that there were no significant differences in motility and viability between group A and group B,while the motility and viability of spermatozoa in group C and group D were significantly increased as compared with those in group A.As compared with group A,the values for DNA integrity parameters including comet rate (CR),tail DNA percentage (TD),tail length (TL) and Oliver tail moment (OTM) were significantly reduced in group C and group D.Group C and group D also displayed a higher proportion of intact acrosome than group A.No significant difference in ROS level was found between group A and group B,while with the increase in acetyl-L-camitine concentration,the ROS level in groups C and D was significantly reduced as compared with that in group A.In conclusion,acetyl-L-camitine at a concentration of 7.5 mmol/L is an effective antioxidant against cryo-damage on post-thawed human spermatozoa.

AIMTo investigate the chemical constituents from Picria fel-terrae Lour.

METHODSColumn chromatography techniques were used to isolate the chemical constituents, physico-chemical constants and spectroscopic analysis were employed for structural elucidation. Results Two triterpenoids named picfeltarraenone I (1) and picfeltarraenin XI (2) were isolated, and their structures were established to be 3,11,22-trioxo-16alpha-hydroxy-(20S,24)-epoxy-cucurbit-5,23-diene (1) and 3,11,22-trioxo-16alpha-hydroxy-(20S,24)-epoxy-cucurbit-5, 23-diene-2beta-O-beta-D-glucopyranoside (2), respectively.

CONCLUSIONCompound 2 is a new compound, the 13CNMR data of compound 1 is reported for the first

Glucosides ; chemistry ; isolation & purification ; Molecular Conformation ; Molecular Structure ; Plants, Medicinal ; chemistry ; Scrophulariaceae ; chemistry ; Triterpenes ; chemistry ; isolation & purification