Gindenosides are the active ingredients of Panax ginseng. 20 (S) -protopanaxadiolocotillol type epimers are the main metabolites of 20 (S) -protopanaxadiol. The previous studies showed that there are stereoselectivity difference in pharmacodynamics and pharmacokinetics between 24R-epimer and 24S-epimer. The purpose of this study was to explore the excretion of the epimers in bile, feces and urine of rat. Liquid chromatography tandem mass spectrometry method has been performed for determination of 24R-epimer and 24S-epimer in bile, feces and urine. 24R-epimer or 24S-epimer was intragastric administered to rats at a single dose of 10 mg x kg(-1). Results showed that after administration the recovery of 24R-epimer and 24S-epimer in feces was 17.69% and 17.09%, respectively, while both of the two epimers were hardly detected in urine. The 48 h cumulative biliary excretion rate of 24R-epimer was 8.01% after administration, while only 1.47% for 24S-epimer. It indicated that there are stereoselectivity in biliary excretion of the epimers with intragastric administration.
Animals ; Bile ; chemistry ; metabolism ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Feces ; chemistry ; Ginsenosides ; chemistry ; pharmacokinetics ; Male ; Mass Spectrometry ; Panax ; chemistry ; Rats ; Rats, Sprague-Dawley ; Stereoisomerism ; Urine ; chemistry

· AIM: To investigate the expression and the possible implication of CD40/CD40L costimulatory molecules in erythema nodosum of patients with Beh(c)et's disease.· METHODS: Sampling was done from erythema nodosum of 5 patients with Beh(c)et's disease and normal skin of 2 healthy individuals. Immunohistochemical staining was performed to examine the expression of CD4, CD8, CD19, CD68, HLA-DR,CD40 and CD40L molecules in the obtained tissues.· RESULTS: Approximately 90% of epidermic cells in erythema nodosum expressed CD40 molecule. In the dermis and subcutaneous tissue, a significantly increased number of CD4+Tcells, CD8+Tcells, CD19+cells, CD68+cells, HLA-DR+cells,CD40L+cells, and CD40+cells were observed in the erythema nodosum as compared with that in normal skin. Double staining showed that CD40L molecules were expressed on 45% of CD4+T cells. CD40 molecules were expressed on 100% CD68+ cells and 59.2% of HLA-DR+cells respectively.· CONCLUSION: A number of CD40/CD40L costimulatory molecules are upreguiated in the erythema nodosum of patients with Behcet's disease.

Along with the increase in fungal infections, Candida albicans prevention and control become the focus of anti-fungal infection at present. This study aims to discuss the effect monomer andrographolide (AG) on C. albicans biofilm dispersion. In the experiment, micro-well plates and medical catheter pieces were used to establish the C. albicans biofilm model. It was discovered by XTT assay and flat band method that 1 000, 500, 250 mg x L(-1) AG could impact the activity of C. albicans biofilm dispersion cells. The morphological structures of residual biofilms on catheter pieces were observed with scanning electron microscopy, which showed that 1 000, 500, 250 mg x L(-1) AG could induce C. albicans biofilm dispersion in a dose-dependent manner, and the dispersed cells were dominated by the yeast phase. According to the real-time fluorescence quantification PCR (qRT-PCR) test, AG could up-regulate HSP90 expression and down-regulate UME6 and PES1 expressions. This study demonstrates that AG could induce C. albicans biofilm dispersion to some extent.
Anti-Inflammatory Agents ; pharmacology ; Biofilms ; drug effects ; Candida albicans ; genetics ; physiology ; ultrastructure ; Diterpenes ; pharmacology ; Dose-Response Relationship, Drug ; Fungal Proteins ; genetics ; Gene Expression Regulation, Fungal ; drug effects ; HSP90 Heat-Shock Proteins ; genetics ; Microscopy, Electron, Scanning ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors

OBJECTIVETo investigate the effects of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) , alone and in combination with fluconazole (FLZ) on FLZ-resistant Candida albicans.

METHODThe minimum inhibitory concentrations (MIC) and sessile MIC80 (SMIC80) of EAHD and FLZ to FLZ-resistant C. albicans were determined by CLSI M27-A3 microdilution method, and the synergy of EAHD combined with FLZ were examined by the checkerboard microdilution assay. Agar plate-method was adopted to observe the rate of antifungal activity according to time-kill curve. HPLC and qRT-PCR were utilized to evaluate the changes of ergosterol content and expressions of related genes, respectively.

RESULTMICs of EAHD ranged from 156 to 1,250 mg · L(-1), those of FLZ from 256 to above 2,048 mg · L(-1) with FICI approximate 0.066 in combination; SMIC80 of EAHD were higher than 1,250 mg · L(-1), SMIC80 of FLZ were higher than 512 mg · L(-1) and up to above 2,048 mg · L(-1). Combination group also showed synergy effect except one group showing addition effect. The results of T-K experiment also confirmed obviously fungicidal effect when treated for 12 h. When compared with control groups, the ergosterol was reduced 85% and 50% in the treatments of combination and EAHD alone by HPLC, respective- ly. The expressions of ERG1, ERG2, ERG6, ERG7 and ERG11 were upregulated, and ACS1, ACS2, MET6 were downregulated when exposed to FLZ. The expressions of the above genes were downregulated by treatment of EAHD. The expressions of ERG2, ERG6, ERG11 were upregulated, while ERG1, ERG7, ACS1, ACS2, MET6 were downregulated in combination group.

CONCLUSIONThe combination of EAHD and FLZ exhibited synergy against FLZ-resistant C. albicans through decreasing the synthesis of ergosterol, and resulting in the breakage of cell membrane.

Antifungal Agents ; pharmacology ; Candida albicans ; drug effects ; growth & development ; metabolism ; Drug Resistance, Fungal ; drug effects ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Ergosterol ; biosynthesis ; Fluconazole ; pharmacology ; Microbial Sensitivity Tests

OBJECTIVETo investigate anti-attachment effect of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) on Candida glabrata.

METHODSerial 2-fold dilution assay was used to determine the minimum inhibitory concentrations MICs of EAHD to C. glabrata. XTT assay was used to evaluate the effect of EAHD against adhesion of C. glabrata. Inverted microscope, scanning electron microscope (SEM) and fluorescein diacetate (FDA) staining were applied to observe the morphological changes of C. glabrata in adhesion. PCR was adopted to inspect the expression of attachment-related genes such as EPA1, EPA6 and EPA7.

RESULTThe MIC of EAHD and fluconazole to C. glabrata were 320 mg · L(-1) and 1 mg · L(-1) respectively. The total cells including budding cells decreased in a dose-dependent manner following EAHD treatment. The expressions of EPA1, EPA6 and EPA7 were downregulated dramatically after EAHD treatment.

CONCLUSIONEAHD could effectively inhibit adherence of C. glabrata.

Acetates ; Candida glabrata ; drug effects ; physiology ; Drugs, Chinese Herbal ; pharmacology ; Fungal Proteins ; genetics ; Lectins ; genetics ; Plant Extracts ; pharmacology

OBJECTIVETo investigate the effect of andrographolide derivative Yanhuning (YHN) on Candida albicans biofilms in rats.

METHODThe rat C. albicans biofilms subcutaneous catheter model was established by intraperitoneally injecting YHN (40, 20, 10, 5, 2.5 mg x kg (-1)), with the FLC (80 mg x kg(-1)) positive group as the control group. After 7 d, CFU counting and XTT assay were used to evaluate the effect of YHN on C. albicans biofllms in vivo. Scanning electron microscopy (SEM) was applied to observe the morphological changes in rat biofilms intervened by YHN. The real-time fluorescence quantification PCR was adopted to detect expressions of C. albicans adhesion-related genes, such as ALS1, ALS3, HWP1, EAP1 and MP65.

RESULTThe YHN group showed much less CFUs on catheter pieces and lower XTT metabolic activity than the blank group, with dosage dependence. SEM also showed that YHN could obviously decrease C. albicans adhesion on subcutaneous catheters in rats. According to qRT-PCR's results, YHN can down-regulate expressions of ALS1, ALS3, HWP1, EAP1 and MP65.

CONCLUSIONYHN could inhibit C. albicans biofilms in rats.

Animals ; Biofilms ; drug effects ; growth & development ; Candida albicans ; cytology ; drug effects ; physiology ; Catheters ; microbiology ; Cell Adhesion ; drug effects ; Diterpenes ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Rats

OBJECTIVETo study the hepatitis B virus (HBV) genotype and its relation to clinical degree and responsiveness to antiviral therapy on hepatitis in order to guide the clinical therapy.

METHODSWe amplified HBV S gene by polymerase chain reaction (PCR), using the second-round PCR product, which was digested by restriction fragment length polymorphism (RFLP). This genotype method was designed under the analysis of the restriction fragment length polymorphism and using the restriction enzymes that identified the genotype-specific sequences. Five restriction enzymes, Hph I , Nci I , Alw I, Ear I and NlaIV, were identified in genotype-specific RFLP from the S gene region. Representative sequences from the S genome region of each HBV genotype were aligned to show the restriction sites by the five restriction enzymes. The amplified S gene nucleotide sequences were sequenced by dideoxy-chain-termination method and the corresponding amino acid sequence was deduced using DNASIS software. Later, they were genotyped by comparing to representative S gene sequences obtained from GenBank. This confirmed the results of RFLP HBV genotyping methods, coincident with that of S gene sequence.

RESULTSGenotypes A, B, C, D were classified in 216 patients with HBV and DNA positive. The results showed that: 1 case (0.46%) of genotype A, 19 cases genotype B (8.8% ), 175 genotype C (81.02%) and 21 genotype D (9.72%). A total of 86 patients in the hospital were divided into either genotype C cases (69) or non-genotype C cases (17).

CONCLUSIONGenotype C was the major genotype in Changchun. Among HBV patients, type C was 80.95%, followed by genotypes B and D. Both hepatocellular carcinoma and liver cirrhosis showed relations with genotype C.

Antiviral Agents ; pharmacology ; Carcinoma, Hepatocellular ; virology ; Drug Resistance, Viral ; Genotype ; Hepatitis B ; drug therapy ; Hepatitis B virus ; classification ; drug effects ; genetics ; Humans ; Liver Cirrhosis ; virology ; Liver Neoplasms ; virology ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length

This study was aimed to investigate the effects of extract from Tibetan medicine Berberis Cortex (TMBC) on expressions of protein kinase C (PKC-β), vascular endothelial growth factor (VEGF) and hypoxia-inducible factor (HIF-1α) in the retina of diabetic rats. Diabetic rats were established by one time intraperitoneal injection of strepto-zocin (STZ). Rats were randomly divided into the model group, low-dose TMBC group, medium-dose TMBC group, high-dose TMBC group, metformin group, calcium dobesilate group, berberine group and the normal control group. Intragastric administration was given. The medication amounts of TMBC in the low-dose, medium-dose and high-dose group were 5, 10, and 20 times to the adult medication dose. In the metformin group and the berberine group, 10 times of the adult medication dose were given. Distilled water was given in the model group and the normal con-trol group. After 6-week intragastric administration, all experimental rats were sacrificed. The expressions of PKC-β, VEGF and HIF-1α were examined by real-time fluorescence quantitative PCR assay and western blot. HIF-1α ex-pression was detected by immunohistochemistry. The results showed that compared with the normal control group, the mRNA and protein expression of PKC-β, VEGF and HIF-1α increased obviously in the retina of diabetic rats ( P<0.01). Compared with the model group, the mRNA expression of PKC-β, VEGF and HIF-1α in the high-dose and medium-dose TMBC group reduced obviously (P< 0.01). The protein expression levels of PKC-β, VEGF and HIF-1α were also obviously reduced (P< 0.05). The expressions of PKC-β, VEGF and HIF-1α in the low-dose TMBC group were obviously reduced (P < 0.05). It was concluded that TMBC can depress the expressions of HIF-1α, PKC-β and VEGF in the retina of diabetic rats, which can be served as a protective effect to prevent progress of di-abetic retinopathy.

The self-designed experimental teaching method is introduced in detail in this paper,including the preparative work before class,discussion of experimental designing proposal,and accomplishment of specific experiment and so on.The teaching method innovations on pathophysiology experiment are very helpful to cultivate the students' ability to solve practical problem and lay the foundation to cultivate talented medical science personal.

Objective To study the effect of background music on spatial long-term learning and memory in rats.Methods Morris' water maze was used to examine the mean latencies and strategies of the rats to the target while playing rock music(RM) and light music(LM).The long-term memory was tested on day 7 after the training sessions.Results During Morris' water maze training,no significant difference was found in the mean latencies and the strategies to the target between RM and LM groups as well as LM and the normal control(NC) groups.But the mean latency in RM group was significantly longer than that in the NC group.The average percentage of straight strategy in RM group was significantly smaller than that in the NC group.The examination of memories,when tested on day 7 after the end of training,indicated that the mean latencies and the strategies to the target as well as the time taken to cross the target in the three groups were not significantly different.Conclusion Background rock music can influence the spatial learning ability,but not the spatial long-term memory.On the other hand,there is no effect in the spatial long-term learning and memory when light background music is played.