OBJECTIVETo extract RNA from Pinellia ternata and lay a foundation for studying the formation mechanism of P. ternata.

METHODBy modifying the method recommended by Guanidinium for extracting total RNA from plant tissues rich in phenolic and polysaccharidic compounds, a simple and convenient method for extraction of total RNA from the tubers, stems and leaves of P. ternate containing abundant polyphenols and polysaccharides was established. High concentrated p-mercaptoethanol was added in the RNA extracted buffer to remove polyphenols, phenol and chloroform were used to eliminate proteins, and isopropanol and sodium acetate were used to precipitate polysaccharides.

RESULTThe A260/A230 value of RNA extracted with improved method were all over 2.0 and the values of A260/A280 were between 1.7 and 2.0. The electrophoresis bands were cleared on agarosegel and integrity of RNA was good.

CONCLUSIONThe results showed that RNA obtained from the tubers, stems and leaves of P. ternate with this method had high purity and quality and could be used in molecular biological research, as DDRT-PCR and reverse Northern blotting analysis directly. This method is simple, economic, stable performance, and has a good repeatability as well as is suitable for extracting total RNA of medicinal plants with high concentrations of phenolics and polysaccharides.

Blotting, Northern ; Pinellia ; genetics ; Plant Leaves ; genetics ; Plant Stems ; genetics ; Plant Tubers ; genetics ; Polymerase Chain Reaction ; RNA, Plant ; isolation & purification

OBJECTIVETo observe the anti-viral therapy effect on HBV reactivation in malignant tumor patients and hepatitis B virus carriers after their cancer chemotherapy.

METHODSThirteen cancer patients but also chronic hepatitis B virus carriers were enrolled in this study. They were randomly put into two groups. Eight patients were put in the therapeutic group. They all had abnormal liver functions induced by the reactivation of HBV after their cancer chemotherapy. Then they were treated with lamivudine. The other 5 cases were treated with lamivudine before their cancer chemotherapy when their serum HBV DNA levels were less than 10(3) copies/ml (preventive therapeutic group). The two groups were followed-up with liver function tests and serum HBV DNA level measurements.

RESULTSAmong the 8 cases of the therapeutic group, 5 cases died of liver failure; cancer chemotherapy was postponed or even terminated in 3 patients due to liver function abnormality and anti-virus treatment was started. In the preventive therapy group, no HBV reactivation was observed in any of the 5 cases.

CONCLUSIONFor HBV carrier cancer patients, an anti-viral therapy before their cancer chemotherapy seems to be very important.

Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; Antiviral Agents ; therapeutic use ; Carrier State ; virology ; Female ; Hepatitis B ; virology ; Hepatitis B virus ; drug effects ; Humans ; Lamivudine ; therapeutic use ; Male ; Neoplasms ; drug therapy ; virology ; Virus Activation ; drug effects

To study the effect of Siwu decoction on the function and expression of P-glycoprotein (P-gp) in Caco-2 cells. The Real-time quantitative poly-merase chain reaction (Q-PCR) was used to analyze the mRNA expression of MDR1 gene in Caco-2 cells. Flow cytometer was used to study the effect of Siwu decoction on the uptake of Rhodamine 123 in Caco-2 cells, in order to evaluate the efflux function of P-gp. Western blotting method was used to detect the effect of Siwu decoction on the P-gp protein expression of Caco-2 cells. Compared with the blank control group, after Caco-2 incubation with Siwu decoction at concentrations of 3.3, 5.0, 10.0 g x L(-1) for 24, 48, 72 h, the mRNA expression of MDR1 was up-regulated, suggesting the effect of Siwu decoction in inducing the expression of MDR1. After the administration with Siwu decoction in Caco-2 cells for 48 h, the uptake of Rhodamine 123 in Caco-2 cells decreased by respectively 16.6%, 22.1% (P < 0.05) and 45.4% (P < 0.01), indicating that the long-term administration of Siwu decoction can enhance the P-gp efflux function of Caco-2 cells. After the incubation of Caco-2 cells with Siwu decoction for 48 h, the P-gp protein expression on Caco-2 cell emebranes, demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp.
ATP Binding Cassette Transporter, Sub-Family B ; genetics ; metabolism ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Caco-2 Cells ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Up-Regulation ; drug effects

Soxhlet extraction is a common method of sample preparation. However, there has been no discussion about the efficiency of Soxhlet extraction from different batches and the factors that cause content fluctuation. In this study, Panax ginseng was selected as a model sample. Soxhlet extraction by means of a water bath, which has always been neglected, was identified as a novel key factor in the poor repeat-ability in different batches of Soxhlet extraction, as it can affect the siphon times and reflux time, which have been positively correlated with the ginsenoside contents. By substituting round bottom flasks in the same column, the relative standard deviation of the most fluctuated compound, ginsenoside Rb1, was decreased from 24.6% to 5.02%. Scanning electron microscopy analysis confirmed that the breakdown of the surface of the ginseng powder in the Soxhlet extraction led to a better dissolution of ginsenosides, indicating that chloroform may promote the extraction of ginsenosides by disrupting the cell structure. Moreover, 70% methanol was regarded as the better solvent for extracting the ginsenosides. Overall, this work offers a practical and effective protocol for improving the accuracy and repeatability of Soxhlet extraction methodology for ginsenosides and other analytes.

Objective To explore the effect of nursing leadership on health education in patients after stroke.Methods Inpatients after first stroke received health education provided by nurses.When the patients went back to out-patient department after discharge, 64 of them were selected as subjects.The Stroke Knowledge Questionnaire (SKQ) and Health Promoting Lifestyle Profile Ⅱ (HPLP Ⅱ) were used.Results Except "Spiritual Growth", the scores of SKQ and other subscales of HPLP Ⅱ were higher than those before stroke.But the standard score of SKQ after stoke was only (67.14±17.18).Conclusions Our health education can effectively enhance the health knowledge and behavior in patients after stroke, but it is not satisfied, it is important to enhance nurses' leadership to improve our health education.

OBJECTIVETo investigate the positive ratio and clinical significance of PreS1Ag and anti-PreS1 in patients with chronic hepatitis B.

METHODS428 patients with chronic HBV infection were collected, these patients were divided into e antigen-positive CHB group, e antigen-negative CHB group, inactive HBsAg carrier group and HBsAg serum conversion group. The difference of positive ratio of PreS1Ag and anti-PreS1 among all groups or between every two groups were analyzed; The relationship of PreS1Ag and anti-PreS1 with HBV M and HBV DNA were also analyzed. SPSS13.0 software was used for statistical treatment. Fourfold table chi-square test or matched-pairs chi-square test was used for enumeration data, and independent sampler t test or rank-sum test was used for measurement data.

RESULTSThe differences of PreS1Ag among four groups were statistically significant (X2=141.7, P<0.05). The positive ratio of PreS1Ag in e antigen-positive CHB group was 95.7%, followed by 82.8% in e antigen-negative CHB group, 13.2% in inactive HBsAg carrier group and 2.2% in HBsAg serum conversion group. The difference of positive ratio of anti-PreS1 between HBsAg seroconversion group and HBsAg positive group was statistically significant (X2=6.919, P<0.05), which indicated that anti-PreS1 had good correlation with HBsAg seroconversion. The average absorbance ratio of PreS1Ag in high viral replication group (179.30) was higher than that in low viral replication group (133.87), statistical significance appeared (Z=-3.86, P<0.05). Though the difference of absorbance ratio of anti-PreS1 between two groups had no statistical significance (P>0.05), descent trend was apparent with virus replication level ascending. We analyzed the concordance of anti-HBs and anti-PreS1 by matched-pairs chi-square test, result showed no statistical significance of detection rate between them, X2=0.262, P>0.05. Serum PreS1Ag, HBeAg or HBcAg in liver tissue in reflecting hepatitis B replication had correlation with HBV DNA (X2=33.840, 24.159, 4.854 in order, P<0.05). Correlation coefficient between PreS1Ag and HBV DNA was higher (r=0.628) than that between HBeAg and HBV DNA (r=0.563).

CONCLUSIONPreS1Ag was more sensitive than HBeAg in diagnosing viral replication in patients with chronic hepatitis B. Anti-PreS1 as protective antibody may be involved in clearance of hepatitis B, positive result indicated recovery of chronic hepatitis B.

Adult ; Female ; Hepatitis B Antibodies ; blood ; immunology ; Hepatitis B Surface Antigens ; blood ; immunology ; Hepatitis B, Chronic ; blood ; immunology ; Humans ; Male ; Middle Aged ; Protein Precursors ; blood ; immunology

OBJECTIVETo understand the current epidemic situation and high risk factors of human immunodeficiency virus (HIV) infection among paid blood donors living in Shandong provincial China Comprehensive Response (CARES) Project Areas.

METHODSAll residents between 20 to 60 years old were selected from one or several counties in August 2003.

RESULTSThere were 19 HIV(+) infections among 661 subjects interviewed. HIV prevalence rate among paid blood donors was higher (3.98%) than that of others (0.48%) while HIV prevalence rate was higher in plasma donors (7.24%) than that in both plasma and full blood donors (2.90%). There was no infections identified in full blood donors. Donors who donated blood during 1994 - 1995 had a higher prevalence rate (7.07%) than those who started donation after 1995 and those began donation before 1993 (0.99%). The rate of condom use was lower among paid blood donors (13.72%) than that of others. There was a lower rate of voluntary HIV testing among paid blood donors than that of others.

CONCLUSIONThe main high risk population of HIV infection were paid donors in CARES Project Areas, whose risk factor was plasma donation and were infected mainly before 1995. Both the rate of condom use and HIV testing were all very low, indicating that they knew little on HIV/AIDS and paid less attention to self-proctection. It is necessary to enhance HIV/AIDS health education and behavior intervention on paid blood donors.

Adult ; Blood Donors ; China ; epidemiology ; Female ; HIV Infections ; epidemiology ; HIV Seroprevalence ; HIV-1 ; Humans ; Incidence ; Male ; Middle Aged ; Plasma Exchange ; adverse effects ; Risk Factors ; Seroepidemiologic Studies

In order to construct an eukaryotic expression vector for gene of multiple myeloma mucin1 (muc1-2vntr) gene and to express it in COS-7 cells in vitro, so to provide the basic material for further research of multiple myeloma DNA vaccine. muc1-2vntr coding gene was used as a research gene and a KOZAK sequence was inserted before the gene Hind III and XbaI restriction sites were inserted before and after the coding gene. Then the whole sequence was synthesized and inserted into pcDNA3.1/myc-his B vector, and the resulted recombinant vector was transformed into E.coil competent cells to get an engineering strain, the recombinant plasmid pcDNA3.1-2vntr/myc-his B identified by restriction analysis and DNA sequencing were transfected into COS-7 cells by liposome-mediated gene transfer method. Finally, fluorescent microscopy was used to assess GFP expression and Western blot analysis using muc1 monoclonal antibody was used to recognize vntr, confirming the expression of vntr. The results showed that the full length of synthesized muc1-2vntr gene, as expected, was 140 bp. Both restriction analysis and DNA sequencing demonstrated that pcDNA3.1-2vntr/myc-his B included the whole translation frame region and muc1-2vntr gene. Furthermore, the fluorescence microscopy proved that the recombinant plasmid had been successfully transfected into COS-7 cells. The expression of mucin-1 protein was observed both in the transfected cell and the cell supernatant by Western blot. It is concluded that the pcDNA3.1-2vntr/myc-his B has been successfully constructed and expressed in COS-7 cells in vitro, which provides the basic material for further researches of mucin-1 function and possible multiple myloma DNA vaccine.
Animals ; Base Sequence ; COS Cells ; Cercopithecus aethiops ; Genetic Vectors ; Molecular Sequence Data ; Mucin-1 ; genetics ; Multiple Myeloma ; genetics ; Plasmids ; Transfection

OBJECTIVEThis study estimated the investment in child development from three aspects-public health, public education, and family investment to establish the level of investment, to provide reference information for government decision making and to provide international comparisons.

METHODSPublic investment in health was measured with macro data related to public health spending and child development in government expenditure. Public education investment was based on basic education data. Family investment evaluation was based on per capita family consumer spending data in different age groups to estimate the input for child development.

RESULTSBoth public health investment level and the proportion of GDP rose for all age groups over time, but the overall investment level was still insufficient. Public investment in children's education has increased year by year, but the trends in all age groups are unbalanced with much lower investment in early childhood education. Private investment in children has increased over the period, but has declined as a percentage of GDP. International comparisons show that Chinas investment in child development is much lower than OECD countries.

CONCLUSIONThe private investment in child development was the main way in China, with public finance contributing only a small proportion. Given the poor international comparisons, the government needs to review the balance of public investment to redirect more towards the development of children under the age of six to their health and education.

Adolescent ; Child ; Child Development ; Child, Preschool ; China ; Female ; Health Expenditures ; Humans ; Infant ; Infant, Newborn ; Male ; Young Adult

OBJECTIVETo establish and characterize imatinib-resistant gastrointestinal stromal tumor (GIST) xenografts. Further provided an ideal experimental platform through the imatinib-resistant GIST xenografts to investigate the mechanism of resistance to imatinib.

METHODSImatinib-resistant GIST cells were injected under the skin of athymic nude mice to establish animal models of human imatinib-resistant GIST. The molecular and histopathologic features of GIST xenografts were also analysed and compared with their counterpart of cell lines.

RESULTSThe xenograft tumor models had been established by subcutaneously injection of GIST cells into nude mice. Immunohistochemistry results showed CD117 expression was positive in GIST-PR2 xenograft tumor, but negative in GIST-R. In GIST-PR1, tumor areas showing rhabdomyoblastic differentiation were presented next to areas with classic GIST morphology. The rhabdomyoblastic component demonstrated consistently positivity for desmin and myogenin, whereas CD117 was completely negative. The mutation profiles of these xenograft tumors were the same as their counterpart of cell lines.

CONCLUSIONSHuman GIST xenografts with mutation in c-kit have been established from imatinib-resistant GIST lines. Those models will enable further studies on mechanisms of resistance, combination therapies and allow testing of novel targeted therapies.

Animals ; Antineoplastic Agents ; pharmacology ; Benzamides ; Cell Differentiation ; Cell Line, Tumor ; Desmin ; metabolism ; Drug Resistance, Neoplasm ; Female ; Gastrointestinal Neoplasms ; genetics ; metabolism ; pathology ; Gastrointestinal Stromal Tumors ; genetics ; metabolism ; pathology ; Humans ; Imatinib Mesylate ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Mutation ; Myogenin ; metabolism ; Piperazines ; pharmacology ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; Pyrimidines ; pharmacology ; Rhabdomyosarcoma ; metabolism ; pathology ; Xenograft Model Antitumor Assays