Objective: To develop a method for separation and purification of tanshinone from Salvia miltiorrhiza by combination of silica gel and high-speed counter-current chromatography (HSCCC). Methods: The crude extract of S. miltiorrhiza was separated by silica gel chromatography and F1 and F2 were obtained. Then, F1 and F2 were separated by HSCCC with a two-phase solvent system composed of petroleum ether-methanol-water (4:3:4:2 and 8:5:8:3), respectively. The lower phase was used as the mobile phase with a flow rate of 2.0 mL/min, while the apparatus rotated at 850 r/min and the eluates were detected at 254 nm. The structures of the target compounds were identified by ESI-MS and NMR. Results: From 80 mg of F1, three compounds with tanshinone I (14 mg), dihydrotanshinone I (22 mg), and tanshinone IIA (26 mg) were obtained. And from 80 mg of F2, dihydrotanshinone (11 mg), trijuganone B (15 mg), and cryptotanshinone (30 mg) were obtained. The purities of these six compounds determined by HPLC were all over 96%, respectively. Conclusion: Combination of silica gel and HSCCC is an efficient method for separation of tanshinone from S. miltiorrhiza.

Objective:To explore the association between the CYP21A2 genotype and the virilization severity in girls with classic 21-hydroxylase deficiency (21-OHD), so as to further the understanding of virilization in females and provide guidance for prenatal diagnosis and genetic counseling. Methods:A total of 23 patients with two X chromosomes (46, XX) who were newly diagnosed with classic 21-OHD in Wuhan Children′s Hospital from August 2010 to March 2019 were included.These patients were divided into 3 groups according to the Prader grades of the degree of external genitalia masculinization.The 17-hydroxyprogesterone (17-OHP) level, androstenedione (AND) level, testosterone (T) level, dehydroepiandrosterone sulphate (DHEAS) level and genotypes were recorded.The gene mutations were divided into the Null group(the enzyme activity was completely impaired), group A(1% of the normal enzyme activity was retained), group B(2% of the normal enzyme activity was retained) and group C(20%-60% of the normal enzyme activity was retained). The correlation between the gene variation of different Prader grades and the corresponding gene groups were analyzed.Results:All 23 girls showed different degrees of external genitalia masculi-nization.There was a significant positive correlation between the Prader grades and the type of gene variation ( rs=0.696, P<0.001). The gene group A and Null group were highly matched with the Prader Ⅳ(the matching rate: 77.8%). Eighty percent of girls with Prader Ⅱ and Prader Ⅲ were classified as the gene group B. The testosterone le-vel of girls with Prader Ⅳ at first diagnosis was significantly higher than that of girls with Prader Ⅱ [4.6(4.0, 15.0)μg/L vs.0.63(0.40, 1.39)μg/L]( χ2=15.117, P<0.05). Conclusions:There is a significantly positive and strong correlation between the degree of external genitalia masculinization and the degree of deficiency of enzyme activity caused by gene variation in girls with typical 21-OHD.It can provide reference for both parents carrying CYP21A2 gene in prenatal diagnosis, genetic counseling and individualized diagnosis and treatment.

OBJECTIVE: To establish an HPLC method for the determination of the contents of the related substances in metoclopramide.METHODS: The sample was separated on C18 column with the mobile phase consisted of acetonitrile-0.02 mol?L-1 phosphonic acid solution(19∶81) at a flow rate of 1.0 mL?min-1.The detection wavelength was set at 275 nm;the column temperature was kept under room temperature and the injected volume was 20 ?L.The contents of the related substances in metoclopramide were computed by self-control method of main constituent.RESULTS: Under the above described chromatographic conditions,metoclopramide was completely separated from its impurities.A good linearity between impurities' peak area and metoclopramide(contrast solution) concentration was achieved when the concentrations of impurities were over the range of 0.05%～4.0%.The lowest detectable limit of metoclopramide was 0.3 ng,and the contents of the related substances were all less than 0.26%.CONCLUSION: The method is convenient,accurate,sensitive and,specific,and it can be used for the determination of the related substances in metoclopramide.

Objective To explore the association of Gly71Arg mutation in gene of bilirubin uridine 5'-diphosphate-glucuronosyltransferase(UGT1A1)and neonatal jaundice in Beijing city Han population.Methods The genotypes and alleles of the Gly71 Arg polymorphism for UGT1A1 gene were identified by polymerase chain reaction-restricted fragment length polymorphism assay in infants of Beijing city Han population of China,including 96 infants with neonatal jaundice[serum bilirubin(307.06?38.5)?mol/L,indirect bilirubin(292.9?35.9)?mol/L] and 101 healthy control infants [serum bilirubin(131.2?42.1)?mol/L,indirect bilirubin(126.3?39.7)?mol/L].The genotypes and allele frequencies of the polymorphism were compared between infants with neonatal jaundice group and healthy infant group(control group).The effect of polymorphism in infants with neonatal jaundice group on serum bilirubin level were analyzed.Results There were significant differences in genotypes distribution in Gly71Arg polymorphism for UGT1A1 gene between the 2 groups(?2=9.47 P=0.002).Compared with control group,neonatal jaundice group had significantly higher Arg allele frequency in the polymorphism for UGT1A1 gene(?2=10.34 P=0.001).There were independent effects of Gly71Arg mutation in the gene on serum bilirubin level in neonatal jaundice group,at the carriers of homozygote of the Arg allele of Gly71Arg polymorphism had higher serum bilirubin levels compared to carriers of heterozygote of the Arg allele of the polymorphism and non-carriers of the Arg allele of the polymorphism(Pa

OBJECTIVETo study the effect of electro-acupuncture (EA) at points along Foot Yangming Channel on metabolite of ulcerative colitis (UC) rats' cerebral cortex and to identify key metabolites by referring to Pi/Wei-brain related theory in Chinese medicine (CM).

METHODSThe UC rat model was set up by dextran sulfate sodium (DSS) method. Male SD rats were randomly divided into the model group and the EA group, 13 in each group. Another 13 rats were recruited as the blank control group. Rats in the blank control group and the model group received no EA. EA was performed at Zusanli (ST36), Shangjuxu (ST37), and Tianshu (ST25) for 5 days by using disperse-dense wave. Then all rats were sacrificed. Their recto-colon and the ileocecal junction were pathomorphologically observed by light microscope and transmission electron microscope (TEM). Cerebral cortexes were extracted. Water-soluble and lipid-soluble brain tissue metabolites were respectively extracted for metabolic research using 1H nuclear magnetic resonance (1H-NMR).

RESULTSEA could obviously improve the general condition of UC model rats, decrease the value of DAI, reduce the infiltration of inflammatory cells in the intestinal tract, stabilize structures such as mitochondria, endoplasmic reticulum and so on (P <0.05). 1HNMR analysis showed that in the model group, contents of glutamic acid, cholesterol, very low density lipoproein (VLDL) in the pallium obviously decreased, while alanine and low density lipoprotein (LDL) significantly increased. After EA, levels of lactic acid, glutamic acid, total cholesterol (TC), and VLDL all increased, and levels of alanine and LDL decreased. All indices were approximate to those of the blank control group.

CONCLUSIONEA at Foot Yangming channel was found to have some effect on metabolites in the brain tissue of UC model rats, which had specific metabonomic material basis and mechanism based on the Pi/Wei-brain related theory.

Acupuncture Points ; Animals ; Cerebral Cortex ; metabolism ; Colitis, Ulcerative ; Electroacupuncture ; Lipids ; Male ; Rats, Sprague-Dawley

Diagnosis, Differential ; Endothelium, Vascular ; metabolism ; pathology ; Female ; Humans ; Hyperplasia ; Jejunal Diseases ; metabolism ; pathology ; surgery ; Jejunum ; blood supply ; pathology ; surgery ; Middle Aged ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism

BACKGROUND:Previous studies have confirmed that ethanol can promote adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), and up-regulate the expression of PPARγ and aP2 in the tumor necrosis factor α (TNF-α) signaling pathway. As a member of the ZIP protein family, Zrt/Irt-like protein 1 (ZIP1) is closely related to bone metabolism and osteogenic differentiation.OBJECTIVE:To study the effect of BMSCs transfected by ZIP1 on TNF-α signaling pathway in the process of adipogenic differentiation.METHODS:The BMSCs from rabbits were isolated and cultured under different concentrations of alcohol (0.03, 0.09,0.15, 0.21 mol/L), followed by transfection by ZIP1 siRNA and ZIP1 expression vector.RESULTS AND CONCLUSION:After culture in alcohol, the expression levels of aP2 and PPARγ proteins were both significantly increased (P < 0.05), and the level of triglyceride was increased in all alcohol groups except for 0.03 mol/L alcohol group (P < 0.05). After siRNA transfection, the expression levels of aP2 and PPARγ as well as the level of triglyceride were increased significantly in all the alcohol groups (P < 0.05); however, ZIP1 transfection decreased the expression levels of aP2 and PPARγ proteins (P < 0.05). To conclude, ZIP1 siRNA could promote the adipogenic differentiation of BMSCs through the activation of TNF-α signaling pathway.

A quick HPLC-ESI-MS/MS method was established for simultaneous determination of three chemical compositions in Usnea, including usnic acid, diffractaic acid, and ramalic acid. The separation was performed on a chromatographic column of Agilent ZORBAX SB-C, (4.6 mm x 250 mm, 5 µm), and the mobile phase was methanol (0.05% formic acid)-0.05% formic acid solution (4 mmol ammonium acetate), with an isocratic elution at a flow rate of 0.8 ml · min⁻¹. Multiple reaction monitoring scanning mode (MRM) was performed combined with the ion switching technology in positive and negative ion switching mode to apply for the quantitative determination. The calibration curves for the above three compounds were linear in corresponding injection amount. Their average recoveries were 95.0%-105.1%, with RSDs of 1.1%-5.2%. The method was simple, rapid, accurate with high repeatability, which could provide a reference for overcalling evaluation the quality of Usnea.
Chromatography, High Pressure Liquid ; methods ; Hydroxybenzoates ; analysis ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tandem Mass Spectrometry ; methods ; Usnea ; chemistry

OBJECTIVETo study the epidemiolgy of respiratory syncytial virus (RSV) infection in children and its relations to climate factors in Hangzhou.

METHODSMonthly positive rate of RSV in pneumonia inpatients and climate factor including mean air temperature, mean relative humidity and rainy days per month were continuously observed for 3 years. Correlation analysis for RSV positive rate and these three climate factors were performed using partial correlation, and regression methods between the positive rate and significant factor was done.

RESULTS13 642 cases were detected and 25.8% showed positive of RSV. The positive rate of RSV in children < or =1 years old, 1-3 years old, > 3 years old were 33.1%, 19.7% and 5.1% respectively with significant difference (chi2 = 763.7, P = 0.000). Rate of RSV infection was increased from December and kept in high level until May or April next year, but were varied at different years. Partial correlations between positive rate and rainy days, mean relative humidity, and mean air temperature per month were 0.32 (P= 0.066), -0.27 (P = 0.117) and -0.83 (P = 0.000) respectively. The regression equation of RSV positive rate and mean air temperature was: RSV positive rate (%) = 52.933 - 1.914 x mean air temperature (degrees C).

CONCLUSIONRSV was one of the main factors causing of pneumonia in children while the highest infectious rate was in children < or =1 year old and infectious rate reduced along with the increase of age. Low air temperature was the main factor affecting the epidemiology of RSV. RSV was prevalent both in spring and winter in Hangzhou area.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Climate ; Female ; Humans ; Humidity ; Infant ; Logistic Models ; Male ; Respiratory Syncytial Virus Infections ; epidemiology ; Seasons ; Temperature

OBJECTIVETo evaluate the viral pathogen of pneumonia in children.

METHODSA total of 13 642 cases of children pneumonia in 3 years were enrolled in this study. Antigens of viral pathogen in respiratory excretion, including respiratory syncytial virus (RSV), type 1, 2 and 3 parainfluenza virus, type A and B influenza virus, and adenovirus were detected by direct immunofluorescence method.

RESULTSViral pneumonia accounted for 34.3% of all cases, including 25.8% cases of RSV, 4.7% of parainfluenza virus, 2.4% of type A influenza virus, 0.2% of type B influenza virus and 1.3% of adenovirus. Coinfection was found in 20 cases, in which 17 cases (85%) were infected with RSV and another virus. Positive rates of RSV in children < or = 1 year, 1 to 3 years, and >3 years were 33.1%, 19.7% and 5.1% with a significant difference (chi(2)(trend)=763.4, P < 0.001). The positive rate of adenovirus in children < or =1 year (0.7%) was significantly lower than that in children aged 1 to 3 years and in children >3 years (2.3% and 2.5%) (all P<0.01). The positive rate of type A influenza virus in children aged 1 to 3 years was higher than that in children < or =1 year (chi(2)=18.2, P<0.01). Type 1 parainfluenza virus was found in 1.2% children aged 1 to 3 years with most prevalence (P<0.05). Infection rates of type 3 parainfluenza in children < or =1 year, 1 to 3 years, and >3 years were 4.7%, 3.2% and 1.4% respectively with a significant difference (chi(2)(trend)=52.4, P<0.01). Although there were some differences of infection rate of RSV in different years, it tended to increase from November to next April with a highest rate of 62.8%. Type 3 parainfluenza virus and Type A influenza virus were almost sporadic while type A influenza virus was epidemic in August 2003 with an infection rate of 15.7%.

CONCLUSIONThe highest infection rate of viral pathogen of pneumonia in children is RSV and the follows are parainfluenza, influenza and adenovirus in turn.

Adenoviridae ; isolation & purification ; Adenovirus Infections, Human ; virology ; Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Orthomyxoviridae ; isolation & purification ; Orthomyxoviridae Infections ; virology ; Paramyxoviridae ; isolation & purification ; Paramyxoviridae Infections ; virology ; Pneumonia, Viral ; virology ; Respiratory Syncytial Virus Infections ; virology ; Respiratory Syncytial Viruses ; isolation & purification