This study is to investigate the effect of Euphorbia humifusa effective fraction (EHEF) on the CYP51 enzyme activity, the lanosterol content and the MEP, SUB gene expression of Trichophyton rubrum. Trichophyton rubrum was treated by EHEF for 7 days at 26 degrees C. The activity of CYP51 enzyme of Trichophyton rubrum in the cell membrane was determined by using ELISA kit, and the lanosterol content was investigated by using high performance liquid chromatography (HPLC), and the MEP, SUB gene expression of Trichophyton rubrum was detected with the reverse transcription polymerase chain reaction (RT-PCR) method. Results showed that EHEF can decrease the membrane CYP51 enzyme activity, and it also can accumulate the fungal lanosterol in a dose-dependent manner, and it also can decrease the gene expression of MEP and SUB. The antifungal mechanism of EHEF may be related to the inhibition on CYP51 enzyme activity, and to the effects on fungal cell membrane ergosterol biosynthesis. It may also play an antifungal effect by inhibiting the MEP, SUB gene expression of fungal proteases.
Antifungal Agents ; isolation & purification ; pharmacology ; Cell Membrane ; drug effects ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Enzyme Activation ; drug effects ; Euphorbia ; chemistry ; Gene Expression Regulation, Fungal ; Lanosterol ; metabolism ; Metalloproteases ; metabolism ; Plants, Medicinal ; chemistry ; Sterol 14-Demethylase ; metabolism ; Subtilisins ; metabolism ; Trichophyton ; drug effects ; genetics ; metabolism

AIM:To investigate the expression and the significance of VEGF-C/D in rat cornea after alkali burning as well as the role of lymphangiogenesis in the high-risk corneal transplantation rejection.
●METHODS:The model of alkali burn corneal was made. Different times corneas were taken to electron microscope for vascularization, and examined the expression of VEGF-C/D and VEGFR-3 in l, 3, 5, 7, 14, 28d. The other rat cornea after alkali burn were divided into four parts to penetrate keratoplasty, containing only blood vessels in the cornea ( group A ) , angiogenesis and lymphangiogenesis ( group B ) , lymphangiogenesis degenerating period ( group C ) , angiogenesis degenerating period ( group D ) . ln addition, there are also normal groups ( group N ) to compare the Rl values and survival time of corneal graft.
●RESULTS: Electron microscopy showed that, when the first 7d rat cornea appeared neovascularization after alkali burn, but not lymphangiogenesis. The occurrence of new blood vessels and lymphatic in 2wk. There were no obvious lymphangiogenesis in 5wk and the angiogenesis gradually subside in 8 wk. The expression of VEGF-C/D and VEGFR-3 in the corneas of rats were up-regulated in the third days after the injury, and reached its peaks at 5d. The average survival time of group N, A, B, C, D were (14.25±0.62)d, (9.35±1.02)d, (5.06±1.13)d, (8.71±0.83) d, (9. 44±1. 05)d after transplant cornea. Compared to the rest of the group, group B plant average survival time significantly shortened (P<0. 05), while compared with group B, the survival time of A, C, D groups were significantly longer (P<0. 05).
● CONCLUSION: VEGF - C/D and VEGFR - 3 are expressed significantly after corneal alkali burn. New lymphatic vessels can accelerate high - risk corneal transplantation immune rejection.

This paper is discussed about course system construction of Microbiology, teaching method, in- struction means and experimental teaching mode. Teaching practice indicated that reform the pattern of Mi- crobiology educational mode can stimulate students’ interest in studying the course, cultivate their inde- pendent ability to solve questions, develop their creative thinking. It is an important way to train high-caliber talents.

Animals ; Chemokine CCL11 ; metabolism ; Guinea Pigs ; Respiratory Mucosa ; metabolism ; Rhinitis, Allergic, Seasonal ; metabolism

OBJECTIVETo study the characteristics and regular pattern of the medicine in common use and combined medication in patients of acute pancreatitis in real world.

METHODCollect the information of 5 433 acute pancreatitis patients in 19 grade IIIA general hospitals in China, analysis by descriptive statistics and association rule.

RESULTIn the 5 433 patients of acute pancreatitis, the glycyrrhizic acid injection and somatostatin are the frequency top used Chinese traditional and western medication. Glycyrrhizic acid injection, somatostatin and insulin are the frequency top used drug combination pattern.

CONCLUSIONThe Chinese and western integrative medicine drug use pattern are accord with the clinical guideline of acute panceatitis. The hepatic and renal function, blood routine and coagulation function should be monitored when the medicines are used.

Adolescent ; Adult ; Aged ; Anti-Inflammatory Agents ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glycyrrhizic Acid ; therapeutic use ; Humans ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Pancreatitis ; drug therapy ; Somatostatin ; therapeutic use ; Young Adult

Objective To obtain the fusion genes of several human orphan G protein coupled receptors (oGPCRs) with Gi1α subtype of G protein and their expression system. Methods The whole open reading frames of GPR45, GPR85, GPR174 and Gilα were cloned by RT-PCR from HepG2 cDNA separately,and the corresponding fusion genes were amplified by overlap extension PCR. Then, the fusion genes-containing pBacmids were successfully constructed with the Bac-to-Bac baculovirus expression system indicated by specific transposition and virus recombination. The insect Sf9 cells were transfected with pBacmid-oGPCRs-Gi1α, and the supernatant containing recombinant virus was harvested. With the supernatant, insect Sf9 cells were infected under an optimized condition (MOI=5, infection time=72 h) and the fusion proteins were prepared and detected by Western blotting.Results The three fusion genes of GPCR45, GPR85 or GPR174 with Gi1α were obtained. The corresponding fusion proteins could be properly prepared in Sf9 cells.Conclusion Human oGPCRs could be fused with Gilα, and the fusion genes could be expressed using the Bac-to-Bac baculovirus expression system in insect Sf9 cells.

OBJECTIVETo investigate whether resveratrol (RES) plays a protective role in hypothermic preserved isolated rat hearts and whether it is mediated by regulation of silent information regulator protein-1 (Sirt-1) expression.

METHODSThe Langendorff model of isolated rat heart was used. After stored in different Celsior solution at 4 degrees C for 9 h, SD rat hearts were randomly divided into 7 groups: blank control group;9 h group (soley hypothermic preservation for 9 h); RES group (3, 10, 30 micromol/L RES treatment plus hypothermic preservation for 9 h ), niacinamide (NAM) group (40 micromol/L NAM added in Celsior solution plus hypothermic preservation for 9 h), RES + NAM group (30 micromol/L RES and 40 micromol/L NAM were added in Celsior solution plus hypothermic preservation for 9 h). The morphological changes of cardiomyocytes were detected by the HE staining with the light microscope. The mRNA and protein expression levels of Sirt-1 were detected by Real-Time PCR and Western blot respectively.

RESULTS(1) Compared with the blank control group, myocardiocytes were injured remarkably in the 9 h group and the Sirt-1 mRNA and protein expression levels were decreased significantly (P < 0.01); (2) Compared with the 9 h group, rat myocardial injury was alleviated gradually in 3, 10, 30 micromol/L RES group and the Sirt-1 mRNA and protein expression levels were increased in a dose-dependent manner (P < 0.05); (3) The above protective effects of RES were attenuated by Sirt-1 inhibitor NAM.

CONCLUSIONRES can protect myocardiocytes from injury caused by long range hypothermic preservation and this protective effect maybe mediated by upregulation of Sirt-1 expression.

Animals ; Cryopreservation ; Heart ; drug effects ; Male ; Organ Preservation ; Rats ; Rats, Sprague-Dawley ; Sirtuin 1 ; metabolism ; Stilbenes ; pharmacology

Objective To explore the influence of obstructive sleep apnea hypopnea syndrome(OSAHS)in children on the secretion of antidiuretic hormone(ADH). Methods Thirty pediatric patients with OSAHS were examined with polysomnography(PSG) and urinary volume was recorded during sleep,and vein blood was sampled in deep sleep to detect the level of ADH in serum using radioimmunoassay technique,which were performed before and after adenotonsillectomy. Among twenty heath children were also detected the secretion of ADH as normal controls. Results After surgery, apnea-hypopnea index(AHI)decreased(from 17.4±2.6 to 3.3 ±1.4,t=27.68,P＜0.001),lowest SaO2 increased(from 0.783 ±0.134 to 0.954 ±0.062, t=6.45, P＜0.001). The level of ADH in OSAHS patients(63.1±35.2)ng/L was much lower than that in health children(85.1±22.2)ng/L(t=2.75,P＜0.01).The serum ADH level in postoperative patients(83.1±21.2)ng/L was increased significantly compared with that of preoperative(t=2.56, P＜0.05), and no statistical difference versus that of health children(t=0.17, P＞0.05). Nycturia volume of preoperative OSAHS children(492±90)ml was significant higher than that of postoperative(332±56)ml or normal controls(346±62)ml(t was 7.85 and 6.43,both P＜0.001). There was no significance in nycturia volume between postoperative group and control group(t=0.77,P＞0.05). Conclusions After adenotonsillectomy in children with OSAHS caused by adenotonsillan hypertrophy. The sleep pattern and ADH secretion could become normal.

OBJECTIVE: To seek a exact method of estimating postmortem interval (PMI). METHODS: This study detected the concentration of magnesium (Mg) and iron (Fe) in vitreous humor of rabbit at 96 h after death and explored the relationship between their concentration and PMI using a method ICP-MS. RESULTS: The concentra-tion of Mg in vitreous humor of rabbit at 48 h after death and Fe in vitreous humor of rabbit at 6-48 h after death were related to PMI significantly. The formulae of the relationship between PMI and Mg concentrations is y = 0.0738x2 + 0.6997x + 11.45 (within 48 h, R2 = 0.9119). The formulae of the relationship between PMI and Fe concentrations is y = 0.0411x2 - 0.3148x + 1.4113 (within 6-48 h, R2 = 0.9594). CONCLUSION The concentration of Mg in vitreous humor of rabbit at 48 h after death and Fe in vitreous humor of rabbit at 6-48 h after death may be as reference indicator to estimate PMI.
Animals ; Iron/analysis* ; Magnesium/analysis* ; Mass Spectrometry/methods* ; Postmortem Changes ; Rabbits ; Time Factors ; Vitreous Body/chemistry*

Objective To investigate the type 2 diabetes mellitus(T2DM)prevalence and related risk factors in adult population with obesity in Tianjin. Methods With stratified cluster randomized sampling, 2888 obese people with BMI≥28 kg/m2, aged 18 years old and over were selected from three urban and three rural regions of Tianjin, in 2006. Information on risk factors was collected with questionnaire through face-to-face interview by trained workers and data on fasting blood glucose(FBG)was collected at the same time. 2hrPPG was tested among the people who' s FBG ≥6.1 mmol/L at the hospital. Prevalence of T2DM was calculated and the distribution of T2DM in the described subgroups and the risk factors analyzed with SPSS software. Results The prevalence of T2DM in adult population with obesity was 11.74%, with females(13.90%)higher than males (8.75%). The prevalence rates of T2DM were statistically different among different groups, classified by age, education, occupation, district and BMI. Results from the univariate and multivariate logistic regression analysis showed that the risk factors of T2DM were age(OR=1.383, 95% CI: 1.254-1 .525)and sex(OR= 1.591,95% CI: 1.230-2.059)while the protective factor was fruit intake(OR=0.867, 95% CI: 0.774-0.971). Conclusion The prevalence of T2DM in adult with obesity was considered to be high. The distribution of T2DM in different subgroups and affecting factors of T2DM in obese adults were different from general population.