In this study, the molecular marker technology of SRAP and ISSR were applied in rapid identification of seeds from eight species of Brassica oleracea L. Firstly, using the genomic DNA of cabbage as template, SRAP and ISSR reaction systems were optimized through testing every factor, respectively, that affects PCR amplification. Then, using the optimized reaction systems, 30 SRAP primer pairs and 15 ISSR primers were applied to amplify genomic DNA of cabbage, savoy, purple cabbage, borecole, cauliflower, broccoli, Brussels sprouts, and kohlrabi The results showed that high polymorphisms were exhibited among the eight species of Brassica oleracea L. by SRAP primer pairs of M3-E5 and M4-E5, as well as ISSR primers of 844 and 888, especially primer 844 which can identify all eight materials efficiently.
Brassica ; genetics ; Nucleic Acid Amplification Techniques ; methods ; Polymorphism, Genetic ; Repetitive Sequences, Nucleic Acid ; Seeds ; genetics

OBJECTIVETo investigate the effect of lipid peroxidation on testosterone (T) in the serum and bcl-2 expression in the Leydig cells of aging male rats.

METHODSThe D-galactose-induced subacute aging male rat model was established and 20 SD rats were randomly divided into two groups of equal number: a D-galactose (D) group and a control (C) group. The activity of superoxide dismutase(SOD) and the level of malondialdehyde (MDA) were examined by spectro-absorptiometer, the expression of bcl-2 by immunohistochemical method, and the concentration of serum T by radio-immunity technique.

RESULTS(1) The activity of SOD in the testis of the D group was (116 +/- 18.09) U/ mg x prot, significantly lower than in the C group [(156 +/- 31.02) U/mg x prot (P < 0.01)]. (2) The level of MDA in the testis of the D group was (1.77 +/- 0.41) nmol/mg x prot, significantly higher than in the C group [(1.19 +/- 0.15) nmol/mg x prot (P < 0.05)]. (3) Serum T in the D group was (2.39 +/- 0.90) nmol/L, significantly lower than in the C group [(8.95 +/- 2.53) nmol/L (P < 0.01)]. (4) The expressions of bcl-2 in the leydig cells of the D and C groups were (35.1 +/- 3.6)% and (49.6 +/- 7.4)% respectively, with statistical difference between them (P < 0.01).

CONCLUSIONLipid peroxidation affects the concentration of serum T and the expression of bcl-2 in the Leydig cells of aging male rats.

Aging ; metabolism ; Animals ; Gene Expression ; Leydig Cells ; metabolism ; Lipid Peroxidation ; Male ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Testosterone ; blood

OBJECTIVETo study the effect of CACNA1H gene mutation G773D on calcium channel function.

METHODSBy the overlap extension PCR we introduced G773D mutation into a human Cav3.2acDNA for constructing the mutant. And then using whole cell clamp technique, we studied the alterations of channel behavior in transfected HEK-293 cells.

RESULTSThere were no difference in kinetics of activation and inactivation of calcium channel between wild type and mutant. However comparing with the wild-type Cav3.2 channel, G773D mutant could increase the calcium current density significantly.

CONCLUSIONCACNA1H gene G773D mutation is able to increase calcium current and neuronal excitability.

Base Sequence ; Calcium Channels, T-Type ; genetics ; physiology ; Cell Line ; Child ; Child, Preschool ; DNA Mutational Analysis ; Epilepsy, Absence ; genetics ; pathology ; physiopathology ; Family Health ; Female ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Patch-Clamp Techniques ; Polymorphism, Single Nucleotide

This study was purposed to investigate the angiogenesis effect of platelet-derived membrane microparticles (PMPs) in chick chorioallantoic membranes (CAM). Thrombin were adopted to activate the platelets and then PMPs were obtained. PMPs were isolated by high speed centrifugation. Flow cytometry (FCM) was adopted to evaluate the efficiency of thrombin to produce PMPs and BCA method was adopted to evaluate the content of PMPs. PMPs were put into CAM and the effects of PMPs on angiogenesis in CAM were observed. The results indicated that after incubation for 72 hours at the concentration of 80 microg/ml PMPs, the vessel nets in a 'spoked-wheel' pattern were shown around mixed fibrous filter membranes, number of vessel ramification was 112.5 +/- 11.31 and ratio of vessel area/CAM area was 6.19 +/- 1.29%, but there were not localized allantoic vessels developing in the control group, the number of vessel ramification and ratio of vessel area/CAM area in control group were 82.6 +/- 8.05 and 1.78 +/- 0.33 respectively, so there was significant difference between PMP and control groups. In above mentioned conditions, the number of vessel ramification and ratio of vessel area/CAM area in VEGF group were 128.4 +/- 10.02 and 7.44 +/- 1.36 respectively, so there was no difference between PMP and VEGF groups. It is concluded that PMPs show promotive effect on the formation of capillaries in chick chorioallantoic membranes.
Animals ; Blood Platelets ; physiology ; Cell-Derived Microparticles ; physiology ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; Humans ; Neovascularization, Physiologic ; drug effects ; Particle Size

OBJECTIVETo study the effect of Songling Xuemaikang Capsule (SXC) on blood pressure of spontaneously hypertensive rats (SHR) and regulatory mechanisms for peroxisome proliferator activated receptor-gamma (PPARgamma).

METHODSTotally 24 10-week-old SHR rats were randomly divided into the blank control group, the Chinese medicine (CM) group, and the Western medicine (WM) group, 8 in each group. Rats in the CM group were administered with SXC at the daily dose of 20 mg/kg by gastrogavage. Those in the WM group were administered with ramipril at the daily dose of 1 mg/kg by gastrogavage. Those in the blank control group were administered with equal volume of normal saline. The blood pressure was measured once per week. The cardiac ultrasound was performed 4 weeks later. Rats were killed and then blood was sampled from abdominal aorta. mRNA expressions of liver PPARgamma and angiotensin II type 1 receptor (AT1R) were detected by fluorescence real-time quantitative PCR. Protein expressions of PPARgamma and AT1R were detected using immunohistochemical assay (SP). The contents of PPARgamma and AT1R were quantitatively analyzed by Western blot.

RESULTSAfter 4 weeks of treatment, the blood pressure decreased in the CM group, showing statistical difference when compared with the blank control group (P < 0.01). CM was inferior to WM in lowering blood pressure. But as a whole, CM was more stable and could maintain blood pressure at a relatively stable level. The cardiac ejection fraction increased in the CM group, showing statistical difference when compared with the blank control group (P < 0.05, P < 0.01). The mRNA and protein expressions of liver PPARgamma were up-regulated in the CM group, showing statistical difference when compared with the blank control group (P < 0.05, P < 0.01). CM could obviously inhibit the AT1R mRNA expression, and down-regulate the protein expression of AT1R, showing statistical difference when compared with the blank control group and the WM group respectively (P < 0.01).

CONCLUSIONSXC decreased blood pressure and improved the cardiac ejection fraction, which might be partially achieved by up-regulating the PPARgamma mRNA expression and protein synthesis, and inhibiting the AT1R mRNA expression and AT1R protein synthesis.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hypertension ; drug therapy ; metabolism ; physiopathology ; Male ; PPAR gamma ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Inbred SHR ; Receptor, Angiotensin, Type 1 ; metabolism

OBJECTIVE: To detect the relationship between leukocytes derived microparticle (CD45 METHODS: The expression of CD45 RESULTS: The percentages of CD45 CONCLUSION High level of CD45
Flow Cytometry ; Humans ; Leukemia, Myeloid, Acute ; Leukocytes ; Neoplasm, Residual ; Prognosis

OBJECTIVE: To investigate the value of CD44 METHODS: Flow cytometry was used to detected the proportion of CD44 RESULTS: The percentage of CD44 CONCLUSION HCD44
Flow Cytometry ; Humans ; Hyaluronan Receptors ; Induction Chemotherapy ; Leukemia, Myeloid, Acute/drug therapy* ; Neoplasm, Residual ; Prognosis ; Spleen

BACKGROUND: Gestational diabetes mellitus (GDM) is usually diagnosed between 24th and 28th gestational week using the 75-g oral glucose tolerance test (OGTT). It is difficult to predict GDM before 24th gestational week because fast plasma glucose (FPG) decreases as the gestational age increases. It is controversial that if FPG ≥5.1 mmol/L before 24th gestational week should be intervened or not. The aim of this study was to evaluate the value of FPG to screen GDM before 24th gestational week in women with different pre-pregnancy body mass index (BMI). METHODS: This was a multi-region retrospective cohort study in China. Women who had a singleton live birth between June 20, 2013 and November 30, 2014, resided in Beijing, Guangzhou and Chengdu, and received prenatal care in 21 selected hospitals, were included in this study. Pre-pregnancy BMI, FPG before the 24th gestational week, and one-step GDM screening with 75 g-OGTT at the 24th to 28th gestational weeks were extracted from medical charts and analyzed. The pregnant women were classified into four groups based on pre-pregnancy BMI: Group A (underweight, BMI < 18.5 kg/m), Group B (normal, BMI 18.5-23.9 kg/m), Group C (overweight, BMI 24.0-27.9 kg/m) and Group D (obesity, BMI ≥28.0 kg/m). The trend of FPG before 24th week of gestation was described, and the sensitivity and specificity of using FPG before the 24th gestational week to diagnose GDM among different pre-pregnancy BMI groups were reported. Differences in the means between groups were evaluated using independent sample t-test and analysis of variance. Pearson Chi-square test was used for categorical variables. RESULTS: The prevalence of GDM was 20.0% (6806/34,087) in the study population. FPG decreased gradually as the gestational age increased in all pre-pregnancy BMI groups until the 19th gestational week. FPG was higher in women with higher pre-pregnancy BMI. FPG before the 24th gestational week and pre-pregnancy BMI could be used to predict GDM. The incidence of GDM in women with FPG ≥5.10 mmol/L in the 19th to 24th gestational weeks and pre-pregnancy overweight or obesity was significantly higher than that in women with FPG ≥5.10 mmol/L and pre-pregnancy BMI <24.0 kg/m (78.5% [62/79] vs. 52.9% [64/121], χ = 13.425, P < 0.001). CONCLUSIONS FPG decreased gradually as the gestational age increased in all pre-pregnancy BMI groups until the 19th gestational week. Pre-pregnancy overweight or obesity was associated with an increased FPG value before the 24th gestational week. FPG ≥5.10 mmol/L between 19 and 24 gestational weeks should be treated as GDM in women with pre-pregnancy overweight and obesity.
Adult ; Blood Glucose ; analysis ; Body Mass Index ; Diabetes, Gestational ; blood ; diagnosis ; epidemiology ; Fasting ; blood ; Female ; Gestational Age ; Glucose Tolerance Test ; Humans ; Incidence ; Pregnancy ; Prevalence ; ROC Curve ; Retrospective Studies

Four new prenylflavonoid glycosides, namely koreanoside H-K (1-4), together with eleven known ones (5-15) were isolated from the leaves of Epimedium koreanum Nakai. Their structures were elucidated by 1D NMR, 2D NMR, HR-ESI-MS, IR and UV. The identification of the sugar moieties was carried out by means of acid hydrolysis and HPLC analysis of their derivatives. It is worth noting that compound 3 and compound 4 were elucidated to contain fucose and quinovose moieties, which were two extremely rare sugar units from the genus Epimedium. The anti-pulmonary fibrosis activity of the new compounds was evaluated using A549 cell line. Compounds 1, 2 and 4 showed significant anti-pulmonary fibrosis activities.
Chromatography, High Pressure Liquid ; Epimedium/chemistry* ; Glycosides/pharmacology* ; Plant Extracts/pharmacology* ; Plant Leaves/chemistry*

Aged ; Biliary Tract Diseases ; surgery ; China ; Humans