This study is to establish an HPLC-DAD-ELSD method for simultaneous determination of 5 flavones and saponins in Rhizoma Anemarrhenae including neo-mangiferin, mangiferin, timosaponin B II, timosaponin B III and timosaponin A III. Samples were analyzed on a Merck Purospher STAR column(4.6 mm x 250 mm, 5 μm). The mobile phase consisted of acetonitrile( A) and 0. 1% formic acid (B) with gradient elution at a flow rate of 1.0 mL · min(-1). The column temperature was set at 40 °C. The DAD detector wavelength was set at 254 nm. The ELSD conditions were as follows: the nebulizing gas flow rate was 2.0 L · min(-1) and temperature of drift tube was 105 °C. The volume was 10 μL. The five compounds were well separated with good linear correlations. The mean recoveries were between 102.0%-104.0%. This method was quick and reliable which provides a foundation for quality control of R. Anemarrhenae.
Anemarrhena ; chemistry ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Drugs, Chinese Herbal ; analysis ; Flavones ; analysis ; Rhizome ; chemistry ; Saponins ; analysis

Objective To learn the quality of life (QOL) and mental health status of the physically disabled people in Zhejiang Province,in order to provide evidence for improving the quality of life and mental health of the physically disabled people.Methods Using questionnaires and interviews to make household surveys of 950 physically disabled people.The questionnaire included the brief quality of life scale (SF-36) and the general health questionnaire (GHQ-28) in the anxiety and depression scale,physical and mental health status.The basic information and other living conditions of physically disabled people were collected through interviews.Results A total of 907 valid questionnaires were recovered,and the effective response rate was 95.47％.Scores of QOL of 907 physically disabled people was 60.89,and anxiety and depression scores were more than 4 points and there was a high score on anxious and depression.The scores of quality of life,anxiety and depression were not statistically significant between males and females(P ＞0.05).There were significant differences in all dimensions of SF-36 and anxiety and depression in different levels of physically disabled people(P ＜ 0.01).There was a significant negative correlation between the quality of life and anxiety and depression in physically disabled people(rs =-0.626、-0.639,P ＜ 0.01).The multiple linear regression showed that physiological function,role physical,physical pain and general health were the influencing factors of anxiety of the physically disabled people,and role physical,physical pain and general health were the influencing factors of depression of the physically disabled people.Condusion The quality of life of physically disabled people were comparably poor,and presented anxiety,depression and other psychological problems.Physical function status directly affects the quality of life and mental health status.

In order to investigate whether cytokine-induced killer (CIK) cells can induce apoptosis of bcr-abl(+) K562 cells, apoptosis of K562 cells and CEM cells induced by CIK cells, etoposide or camptothecin was detected with flow cytometry DNA assay. RT-PCR showed that K562 cells expressed the bcr-abl fusion gene, K 562 cells, K562 cells/etoposide or K562 cells/camptothecin groups showed no sub-G(1) peak. K562 cells/CIK cells group showed sub-G(1) peak (38.1%). CEM cells showed no sub-G(1) peak. CEM cells/camptothecin or CEM cells/etoposide groups showed sub-G(1) peak (23.5% or 32.3% respectively). CEM cells/CIK cells group showed sub-G(1) peak (45.4%). Etoposide or camptothecin did not induce apoptosis of K562 cells. CIK cells induce apoptosis of K562 cells. Bcr-abl fusion gene prevented apoptosis induced by etoposide or camptothecin, but did not prevent apoptosis induced by CIK cells. This property may support the observed adoptive immunologic effect of allogeneic bone marrow transplantation and donor lymphocyte transfusions of CML case relapsing after allogeneic bone marrow transplantation.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; immunology ; Camptothecin ; pharmacology ; Coculture Techniques ; Cytotoxicity, Immunologic ; Etoposide ; pharmacology ; Flow Cytometry ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; K562 Cells ; drug effects ; immunology ; metabolism ; Killer Cells, Lymphokine-Activated ; cytology ; immunology

BACKGROUNDAspermia caused by exogenous testosterone limit its usage in late-onset hypogonadism (LOH) patients desiring fertility. Saikokaryukotsuboreito (SKRBT) is reported to improve serum testosterone and relieve LOH-related symptoms. However, it is unclear whether SKRBT affects fertility. We aimed to examine the effects of SKRBT on spermatogenesis and fertility in aging male mice.

METHODSThirty aging male mice were randomly assigned to three groups. Mice were orally administered with phosphate-buffer solution or SKRBT (300 mg/kg, daily) or received testosterone by subcutaneous injections (10 mg/kg, every 3 days). Thirty days later, each male mouse was mated with two female mice. All animals were sacrificed at the end of 90 days. Intratesticular testosterone (ITT) levels, quality of sperm, expression of synaptonemal complex protein 3 (SYCP3), and fertility were assayed.

RESULTSIn the SKRBT-treated group, ITT, quality of sperm, and expression of SYCP3 were all improved compared with the control group (ITT: 85.50 ± 12.31 ng/g vs. 74.10 ± 11.45 ng/g, P = 0.027; sperm number: [14.94 ± 4.63] × 106 cells/ml vs. [8.79 ± 4.38] × 106 cells/ml, P = 0.002; sperm motility: 43.16 ± 9.93% vs. 33.51 ± 6.98%, P = 0.015; the number of SYCP3-positive cells/tubule: 77.50 ± 11.01 ng/ml vs. 49.30 ± 8.73 ng/ml, P < 0.001; the expression of SYCP3 protein: 1.23 ± 0.09 vs. 0.84 ± 0.10, P < 0.001), but fertility was not significantly changed (P > 0.05, respectively). In the testosterone-treated group, ITT, quality of sperm, and expression of SYCP3 were markedly lower than the control group (ITT: 59.00 ± 8.67, P = 0.005; sperm number: [4.34 ± 2.45] × 106 cells/ml, P = 0.018; sperm motility: 19.53 ± 7.69%, P = 0.001; the number of SYCP3-positive cells/tubule: 30.00 ± 11.28, P < 0.001; the percentage of SYCP3-positive tubules/section 71.98 ± 8.88%, P = 0.001; the expression of SYCP3 protein: 0.71 ± 0.09, P < 0.001), and fertility was also suppressed (P < 0.05, respectively).

CONCLUSIONSKRBT had no adverse effect on fertility potential in aging male mice.

Aging ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Fertility ; drug effects ; Hypogonadism ; drug therapy ; Male ; Mice ; Nuclear Proteins ; analysis ; Sperm Count ; Sperm Motility ; drug effects ; Spermatogenesis ; drug effects ; Testis ; drug effects ; pathology ; Testosterone ; blood

OBJECTIVETo study the mechanism of how iron-regulatory protein (hepcidin) affect iron overload in alcoholic liver disease (ALD).

METHODSThirty male wistar rats were randomly divided into 3 groups: Lieber-Decarli liquid without alcohol group (control group), Lieber-Decarli liquid with alcohol (alcohol group) and hepcidin intraperitoneally injected group (hepcidin group), each rat was fed for 6 weeks. The Serum concentration of Alanine Aminotransferase (ALT), Aspartate Amino Transferase (AST), Iron, Total Iron Binding capacity (TIBC), Ferritin, Malonyl Dialdehyde (MDA) and Hepcidin were determined. Hepatic tissue was examined by hematoxylin and eosin staining, prussian blue iron staining and immunohistochemistry staining.

RESULTS(1) Serum concentration of ALT in control group, alcohol group and hepcidin group were (25.2 ± 4.6) U/L, (37.9 ± 14.3) U/L and (40.9 ± 14.1) U/L (F = 4.907, P < 0.05), respectively. Serum AST among three groups were (32.3 ± 13.4) U/L, (55.0 ± 18.6) U/L and (48.3 ± 26.0) U/L (F = 3.742, P < 0.05), respectively. The secretions of ferritin were (224.72 ± 85.49) ng/ml, (345.59 ± 124.75) ng/ml and (339.47 ± 138.47) ng/ml (F = 3.539, P < 0.05). The serum concentrations of TIBC were (147.30 ± 31.98) μmol/L, (148.04 ± 58.74) μmol/L and (143.28 ± 37.38) μmol/L (F = 1.209, P > 0.05), respectively. The serum concentrations of iron were (55.64 ± 13.32) μmol/L, (60.37 ± 25.89) μmol/L and (49.77 ± 17.64) μmol/L (F = 0.651, P > 0.05), respectively. The serum concentration of MDA were (5.84 ± 2.17) nmol/ml, (6.51 ± 2.23) nmol/ml and (4.27 ± 2.68) nmol/ml (F = 2.782, P > 0.05), respectively. The serum concentration of Hepcidin were (155.96 ± 44.91)ng/ml, (124.11 ± 31.98) ng/ml and (114.96 ± 25.81) ng/ml (F = 3.839, P < 0.05), respectively. (2) Significant fat change observed in the liver of alcohol group. The positive granulations of iron staining were (0.8 ± 1.0), (1.2 ± 1.6) and (1.1 ± 1.1) (F = 0.254, P > 0.05), respectively. No differences found of liver iron express among the three groups. Intraperitoneal injection of hepcidin increased hepcidin expression in liver which was inhibited by alcohol (F = 4.139, P < 0.05).

CONCLUSIONSALD rats with lower hepcidin expression in liver can result in iron metabolism disorder. Ectogenic hepcidin can protect liver against alcohol damage by inhibiting lipid peroxidation.

Alanine Transaminase ; blood ; Animals ; Antimicrobial Cationic Peptides ; metabolism ; Hepcidins ; Iron-Regulatory Proteins ; metabolism ; Liver ; metabolism ; pathology ; Liver Diseases, Alcoholic ; metabolism ; pathology ; Male ; Rats ; Rats, Wistar

OBJECTIVETo explore the molecular mechanism of exocrine immune inflammatory injury of Sjögren's Syndrome and the intervention of Banxia Qinlian Decoction (BQD).

METHODSTotally 18 female NOD mice were randomly divided into the model group, the positive drug group, and the BQD group, 6 in each group. Six female BALB/c mice were recruited as a blank control group. Mice in the blank control group and the model group were gavaged with deionized water at the daily dose of 0.1 mL/10 g body weight. Tripterygium Tablet was administered by gastrogavage to mice in the positive group at the daily dose of 10 mg/kg. BQD was administered by gastrogavage to mice in the BQD group at the daily dose of 60 g crude drugs/kg. After 12 weeks of medication, mice were sacrificed. Their eyeballs were excised and blood collected. Tissues of bilateral parotids and submandibular glands were kept. mRNA transcriptional levels of IL-17, IL-6, type 3 muscarinic acetylcholine receptors (M3R), aquaporin protein-5 (AQP5) were detected by RT-PCR. Expression levels of M3R and AQP5 protein were detected by Western blot. Protein expression levels of IL-17 and IL-6 were detected by ELISA.

RESULTSCompared with the normal group, mRNA transcriptional levels and protein expression levels of IL-17, IL-6, M3R, and AQP5 were significantly up-regulated in the model group (P < 0.01). Compared with the model group, mRNA transcriptional levels and protein expression levels of IL-17, IL-6, M3R, and AQP5 were significantly down-regulated in the positive drug group and the BQD group with statistical difference (P < 0.01, P < 0.05). Compared with the BQD group, mRNA-transcriptional levels of IL-17, IL-6, and M3R, as well as M3R and AQP5 protein expression levels were significantly down-regulated in the positive drug group (all P < 0.01).

CONCLUSIONThe molecular mechanism of BQD in inhibiting SS exocrine neurotoxic injury might be possibly related to regulating Th17/IL-17 immune inflammatory way.

Animals ; Aquaporin 5 ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Interleukin-17 ; metabolism ; Interleukin-6 ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Inbred NOD ; Sjogren's Syndrome ; drug therapy ; immunology ; Submandibular Gland ; Th17 Cells ; Up-Regulation

A new aporphine alkaloid (1), together with five known analogues (2-6), has been isolated from the branch of Litsea greenmaniana by using various chromatographic techniques. Their structures were identified by spectroscopic data analysis ( MS, IR, 1D and 2D NMR) as 2,9-dihydroxy-1,10-dimethoxy-4,5-dihydro-7-oxoaporphine (1), laurotetanine (2), N-methyllaurotetanine (3), isodomesticine (4), isocorydine (5), and norisocorydine (6). Compound 1 was a new compound, and compounds 2-6 were obtained from this plant for the first time.
Alkaloids ; chemistry ; Aporphines ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Litsea ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo study the expression level and significance of glucose transporter 1 (Glut-1) in normal breast tissue, adenosis, adenoma and breast carcinoma.

METHODSA total of 147 cases of female breast tissue samples, including 92 cases of invasive ductal carcinoma, 26 cases of breast fibroadenoma, 24 cases of breast adenosis and 5 cases of normal breast tissues, were collected for quantitative detection of the expression of Glut-1 protein by immunohistochemistry (EnVision method) and Western blot, and its mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSIn normal breast tissue and benign lesions of the breast, Glut-1 was undetectable or only weakly detectable in cytoplasm of ductal and acinar epithelia. In contrast, the intensity of Glut-1 staining was significantly higher in invasive ductal carcinomas (P = 0.0002) with protein expression predominantly in cellular membrane and lesser in cytoplasm. Western blot and RT-PCR analyses showed that the expression of Glut-1 protein and mRNA were significantly increased in invasive ductal carcinoma than fibroadenoma (P =0.001 for protein; P <0.05 for mRNA) and adenosis (P =0.001 for protein; P < 0.05 for mRNA). There was a significant difference among groups (P = 0.0002 for protein; P = 0.0001 for mRNA).

CONCLUSIONSGlucose transport activity, as indicated by Glut-1 protein and its mRNA expression, significantly increases in breast carcinoma than non-cancerous lesions. The over-expression of Glut-1 in breast carcinoma is tightly coupled with tumor cell proliferation, invasion and metastasis, implying that Glut-1 may serve as a new marker in the early diagnosis and prognostication of breast malignancy as well as a new therapeutic target.

Breast Neoplasms ; metabolism ; Carcinoma, Ductal, Breast ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Glucose ; physiology ; Glucose Transport Proteins, Facilitative ; genetics ; metabolism ; Glucose Transporter Type 1 ; genetics ; metabolism ; Humans ; Prognosis

OBJECTIVETo construct the body mass index (BMI) reference data and curves for Chinese children and adolescents from birth to 18 years of age.

METHODSData from two national representative cross-sectional surveys which were The National Growth Survey of Children under 7 years in the Nine Cities of China in 2005 and The Physical Fitness and Health Surveillance of Chinese School Students in 2005. Height (length was measured for children under 3 years) and weight data of 93,702 urban healthy children from nine cities/provinces used to calculate the BMI. The LMS method was used to smooth the BMI, with estimates of L, M, and S parameters, values of percentile and Z-score curves which were required were calculated, and then standardized growth charts were generated. Adult cut-offs for overweight and obesity at 18 years was used to study the cut-offs for children 2 to 18 years of age.

RESULTSThe smoothed percentiles and Z-scores reference data and curves of BMI-for-age for boys and girls aged 0-18 years were made out respectively. BMI cut-off values for overweight and obesity for children from 2 to 18 years of age were also defined. At 18 years, the BMI values are equivalent to the overweight cut-off (> or =24 kg/m2) and obesity cut-off (> or =28 kg/m2) for Chinese adults. Comparison with the reference of the WHO and 2000 CDC for the United States, there were some difference among them, at the 97th percentile curve there was a big difference between Chinese and U. S. adolescents. On the whole, the China BMI curve for boys was higher than the new WHO curve and lower than 2000 CDC at 97 percentile curve, but the China BMI curve for girls was lowest among the three curves. There was also significant difference between China and Japan BMI values at 97 percentile curve.

CONCLUSIONBMI growth curves are very useful in child growth monitoring and nutritional surveillance, discovering overweight and obesity. The BMI growth charts are recommended for use in pediatric clinic and public health service.

Adolescent ; Body Height ; Body Mass Index ; Body Weight ; Body Weights and Measures ; standards ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Obesity ; epidemiology

OBJECTIVETo construct the height and weight growth charts for Chinese children and adolescents from birth to 18 years for both clinical and preventive health care uses.

METHODSData from two national representative cross-sectional surveys which were The National Growth Survey of Children under 7 years in the Nine Cities of China in 2005 and The Physical Fitness and Health Surveillance of Chinese School Students in 2005. The data from 94,302 urban healthy children were used to set up the database of length/height (length was measured for children under 3 years) and weight. The LMS method was used to smooth the growth curves, with estimates of L, M, and S parameters, values of percentile and Z-score curves which were required were calculated, and then generated standardized growth charts.

RESULTSThe 3rd, 10th, 25th, 50th, 75th, 90th, 97th smoothed percentiles curves and -3, -2, -1, 0, +1, +2, +3 Z-scores curves of weight-for-age, length/height-for-age for boys and girls aged 0-18 years were made out respectively. Comparison with the new WHO growth charts and 2000 CDC growth charts for the United States, the results showed that there was some big difference in weight and height among the three growth charts. For boys under 15 years of age and girls under 13 years of age, the China curves are slightly higher than WHO and CDC curves, but after those ages, the China curves fall behind and the difference became larger as age progresses. At the age of 18 years, the Chinese children are 3.5 cm shorter in boys and 2.5 cm shorter in girls as compared with the U. S. children. The difference in weights are very large for the school children, especially in girls. The weight of Chinese boys was 5.9 kg less than that of the U. S. boys at 18 years, and the difference was much bigger in girls, the weight of U.S. girls between 8 to 18 years was 4.1-20.5 kg more than that of Chinese girls at the same age range.

CONCLUSIONThe new growth charts of height and weight were based on national survey data and therefore are recommended as the China national growth standards for use in pediatric clinics and public health service. Application of the charts will promote child growth monitoring, discovering early growth disorder, and will be useful to diagnosis of diseases and assessment of therapeutic effects.

Adolescent ; Body Height ; Body Mass Index ; Body Weight ; Body Weights and Measures ; standards ; Child ; Child, Preschool ; China ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Reference Standards