Objective: The aim of this study was to explore the pharmacokinetics of ginsenoside Rg3 and its deglycosylated metabolite, ginsenoside Rh2 in lincomycin-induced gut microbiota dysbiosis rats after ig administration of ginsenoside Rg3. Methods: An LC-MS/MS analytical method was developed and validated to detect ginsenoside Rg3 and Rh2 in plasma of rats. The method was validated by specificity, linearity, lower limits of quantification (LLOQ), precision, accuracy, matrix effect, recovery, and stability. Lincomycin (orally, 5 000 mg/kg, 7 d) was selected to induce gut microbiota dysbiosis. The fecal moisture contents and the β-D-glucosidase activity were also assessed in this study. And the plasma samples were collected and analyzed after ig administration of ginsenoside Rg3 (20 mg/kg). Results: The results indicated that this method could be used for the determination of the concentration of ginsenoside Rg3 and Rh2 in plasma of rats. The fecal moisture content in rats treated with lincomycin was significantly increased (P < 0.01) and the β-D-glucosidase activity was decreased (P < 0.01) compared with the control rats. The AUC0~∞ and Cmax in gut microbiota dysbiosis rats were increased, while the AUC0~t and Cmax of its active metabolite, ginsenoside Rh2 were significantly decreased (P < 0.01) compared with normal rats. Conclusion: The pharmacokinetic profile of ginsenoside Rg3 and Rh2 is changed in gut microbiota dysbiosis rats, which partly relates to the decreased β-D-glucosidase activity.

In order to find the characteristics of two members of gene family of squaleneexpoxidase (SE) , a quantitative real time PCR method was developed to analyze the expression of Eleutherococcus senticosus SE1 and SE2 gene from different growth periods and in different organs. The result indicated that all the expression of SE2 more than SE1 in the whole growth period and organs of E. senticosus. And in the whole growth period, expression of SE1 showed a low-high-low characteristic. Both expression of SE2 and growth period showed the same trend. The lowest content of the expression was in the roots. SE1 expression have been improved more than SE2 when treated with MeJA. The expression of E. senticosus SE1 and saponins content had significantly positive correlation (P < 0.05) and the correlation coefficients was 0. 858, while the correlation was not significant for SE2. That indicated that SE1 played a key enzyme gene in the biosynthesis of triterpenoidsaponins
Eleutherococcus ; chemistry ; enzymology ; genetics ; growth & development ; Gene Expression Regulation, Plant ; Peroxidase ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Saponins ; analysis ; metabolism ; Transcriptome

OBJECTIVETo investigate the gene mutation in the areas of pre core/core (Pre C/C) and basic core promotor (BCP) of HBV DNA and its clinical significance.

METHODSThe nt 1 735-1 965 segment of HBV DNA was amplified with PCR in 54 cases with chronic hepatitis B and 10 cases with post-hepatitis cirrhosis. Then the PCR product was sequenced.

RESULTSThere were 168 site mutations in 48.5% (33/68) cases with hepatitis B. The first ten mutation sites were nt 1 764 (58.8%), 1 762 (48.5%), 1 799 (21.0%), 1 766 (14.7%), 1 896 (13.2%), 1 754 (8.8%), 1 899 (8.8%), 1 768 (7.4%), 1 814 (7.4%) and 1 913 (7.4%). Three rare mutations of nt 1907, 1 922 and 1 923 were also detected. The mutations of nt 1 896, 1 764 and 1 762 were found in 16.7%, 35.2% and 35.2% of chronic hepatitis, and in 30.0%, 60.0% and 60.0% respectively of post-hepatitis cirrhosis cases. There was statistical significance between the two groups (P < 0.01).

CONCLUSIONThe mutations in the areas of Pre C/C and BCP of HBV DNA might possibly be associated with liver fibrosis. There are many mutation sites in HBV DNA and mutation occurs frequently, therefore gene sequencing is helpful to the design of gene chip and to clinical application.

DNA, Viral ; blood ; genetics ; Gene Frequency ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; blood ; pathology ; virology ; Humans ; Mutation ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length

OBJECTIVE The aim of this study was to develop and validate a LC-MS∕MS method to simultaneously determi-nation of Scutebarbatine A and Scutebarbatine B in rat plasma and applicated this method to pharmacokinetic study of Scutebar-batine A and Scutebarbatine B after oral administration of Scutellaria barbata extract.METHODS Plasma samples were pre-pared with ethyl acetate using liquid-liquid extraction.Then samples were separated by using BDS Hypersil C1 8(50 mm×2.1 mm,2.4 μm)with gradient elution program and oven temperature was set at 40 ℃.And the mobile phase was consisted with acetonitrile∕water(0.05% formic acid) at a flow rate of 0.25 mL∕min.Determination was carried out on a tandem mass spec-trometer in positive ion mode using a multiple reaction monitoring(MRM)via a electrospray ionization(ESI)interface.After oral administration of 500 mg∕kg Scutellaria barbata extract,plasma samples were collected and analyzed by using a DAS soft-ware to analyze to the pharmacokinetic parameters.RESULTS Scutebarbatine A and Scutebarbatine B were liner at the range of 0.5 12~258.0 ng∕mL,0.482~241.0 ng∕mL,respectively with a r 2 larger than 0.994.Accuracy,precious,extraction effi-ciency,matrix effects and stability study were all meet the requirement of the bioanalytical method.The AUC(0-t) for Scutebar-batine A and Scutebarbatine B were(88.69±12.4)μg∕L.h,(57.09±9.84)μg∕L.h,respectively.And the Cmax for Scutebar-batine A and Scutebarbatine B were(10.22±1.31)ng∕mL and (6.27 ±0.80)ng∕mL.CONCLUSION The method was sim-ple,accurate,and sensitive,which could be used for the pharmacokinetic study of Scutebarbatine A and Scutebarbatine B after oral administration of Scutellaria barbata extract.

OBJECTIVETo establish a reliable model for drug screening and therapy by culturing rat femoral head and inducing cartilage degeneration quickly in vitro.

METHODSThe femoral heads from the same SD rats of two-month old were divided into control group and experimental group respectively. They were cultured with DMEM medium plus 10% fetal bovine serum or DMEM medium plus 10% fetal bovine serum plus 50 ng/ml IL-1β for three days. Femoral heads were fixed in 4% paraformaldehyde, decalcified, dehydrated, embedded in paraffin and cut into slices. Specimens were stained with Toluidine blue and Safranine O-Fast Green FCF. The protein expression levels of type II collagen, MMP13, Sox9 and ADAMTS5 were analyzed by immunofluorescence.

RESULTSBoth the Toluidine blue and Safranine O staining were pale in the margin of femoral heads which were stimulated with IL-1β for three days compared to that in control group. The Fast Green FCF staining was positive at the edge of the femoral head in experimental group, which indicated that cartilage became degenerated. The expression levels of both type H collagen and Sox9 were decreased significantly while the expression levels of MMP13 and ADAMTS5 were increased in experimental group.

CONCLUSIONThe model of cartilage degeneration is established by culturing and inducing the degeneration of the femoral heads quickly in vitro.

Animals ; Cartilage Diseases ; genetics ; metabolism ; Collagen Type II ; genetics ; metabolism ; Disease Models, Animal ; Femur Head ; metabolism ; Humans ; In Vitro Techniques ; Interleukin-1beta ; genetics ; metabolism ; Male ; Matrix Metalloproteinase 13 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; SOX9 Transcription Factor ; genetics ; metabolism

OBJECTIVETo study the effects of different fertilization, measure of breeding seedling, breeding and sowing density on seedlings of Psoralea corylifolia.

METHODThe combination of field sowing and indoor test was applied.

RESULTAll agricultural measures showed positive effect on the growth and development of seedling of P. corylifolia.

CONCLUSIONThe suitable sewing time should be the first and second ten days in March. The suitable sowing density is 150 kg x hm(-2). The combined of application of organic manure and chemical fertilizer is better than using one of them alone.

Agriculture ; methods ; standards ; Animals ; Breeding ; Fertilizers ; Manure ; Plants, Medicinal ; anatomy & histology ; growth & development ; Psoralea ; anatomy & histology ; growth & development ; Quality Control ; Seasons ; Seedlings ; anatomy & histology ; growth & development

Objective:To investigate the effects of estradiol on ~(60)Co?-ray induced apoptosis of bone marrow hematopoietic cells of mice,and to discuss the related anti-irradiation mechanism.Methods:KM mice were randomly divided into 3 groups(15 mice/each group):control group(without radiation),pure radiation group and estradiol+radiation group(ER group).The pure radiation group was irradiated by 4.0 Gy?-ray at a dose rate of 1.15Gy/min;the ER group was administered with 0.1 mg estradiol(IM)at 10 days before 4.0 Gy?-ray radiation;and the control group received no special treatment.The apoptotic DNA segments of bone marrow hematopoietic cells were analyzed by DNA agarose gel electrophoresis;flow cytometry was used to examine the apoptosis rate of cells and expression of Fas and Bcl-2 at 4 h,8 h,and 12 h after irradiation.Results:Eight hours after radiation,the apoptotic DNA segments were obviously increased and apoptotic DNA ladder appeared,which was not seen in the other 2 groups.The apoptosis rate of bone marrow hematopoietic cells in ER group was significantly lower than that in the pure radiation group at 4,8,and 12 h after irradiation(P

BACKGROUND:Our previous work has demonstrated that bone marrow mesenchymal stem cels (BMSCs) transplantation can improve the heart function of rats with myocardial infarction. However, the overal efficacy is not satisfactory. OBJECTIVE: To adopt pioglitazone as a peroxisome proliferator-activated receptor gamma (PPAR-γ) agonist combined with BMSCs transplantation therapy, thereby further improving cardiac function of rats with myocardial infarction as wel as investigating the relevant mechanisms. METHODS:Twenty Sprague-Dawley rats with myocardial infarction were induced by the left anterior descending coronary artery ligation. The animals were randomized into two groups: BMSCs and BMSCs+pioglitazone. Two weeks later, al the animals received the injection of BMSCs labeled with PKH26 in PBS into the local infarct zone, and then pioglitazone (3 mg/kg/d) was given by the oral gavage for 2 weeks in the BMSCs+pioglitazone group after the cel transplantation. After 2 weeks of cel transplantation, cardiac functions were evaluated by echocardiography. The expressions of PPAR-γ, Connexin 43 and molecules in TGF-β1/SMAD signaling pathway were examined in different areas of the left ventricle from each harvested heart using immunofluorescent staining, western blot assay and qRT-PCR. RESULTS AND CONCLUSION:There were no differences in the baseline parameters of cardiac function between the two groups. At 2 weeks after cel transplantation, the left ventricular internal diameter at end-diastole, left ventricular internal diameter at end-systole and left ventricular ejection fraction were significantly improved in the BMSCs+ pioglitazone group; the expressions of PPAR-γ and Connexin 43 were distinctly increased in different zones of the left ventricle; the levels of TGF-β1, SMAD2 and SMAD3 were obviously attenuated in the infarct zone and border zone. The above-mentioned findings suggest that pioglitazone, a PPAR-γ agonist, can enhance BMSCs potential in improvingthe heart function after myocardial infarction, and PPAR-γ may elevate the expression of Connexin 43via the blockade of the TGF-β1/SMAD signaling pathway in the procedure.

Global Health ; History, 20th Century ; Humans ; Influenza A Virus, H2N2 Subtype ; genetics ; isolation & purification ; Influenza A Virus, H3N2 Subtype ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; history ; mortality ; virology

OBJECTIVETo clarify the relationship between hepatitis B virus (HBV) infection and IgA nephropathy (IgAN).

METHODSHBV antigen (HBAg) in renal tissues of the patients with IgAN was detected by immunohistochemical technique, the carrier status and localization of HBV DNA in renal tissues were determined by Southern blot analysis and in situ hybridization.

RESULTSSerum HBsAg was detected in 18 of the 100 patients with IgAN (18%), HBAg was detected in 31 of 100 patients (31%) in their renal tissue and in 20 of 31 patients (65%) in their glomeruli, and both HBsAg and HBcAg were detected in 10 of 31 patients (32%), respectively. HBcAg was also found in tubular epithelia (45%, 14/31) and renal interstitium (6%, 2/31), respectively. Five of six cases were proved to be positive of integrated-form HBV DNA in their renal tissue by Southern blot analysis. In situ hybridization demonstrated that HBV DNA was 8/8 and 6/8 positive in their renal tubules and glomeruli of all eight specimens, localized in the nucleus of tubular epithelial cells, glomerular mesangial cells, as well as infiltrated interstitial lymphocytes.

CONCLUSIONHBV infection closely related with IgAN and HBV infection might be involved in pathogenesis of IgAN.

Adolescent ; Adult ; Female ; Glomerulonephritis, IGA ; complications ; virology ; Hepatitis B ; complications ; Hepatitis B Antigens ; analysis ; Hepatitis B virus ; isolation & purification ; Humans ; Male ; Middle Aged