OBJECTIVEDetection HBV DNA among HBsAg negative children who have been vaccined at birth, in order to improve the evaluation of the indicator for HBV DNA infection.

METHODSSelection HBsAg negative children who have been vaccined at birth and then detection HBV DNA from sera using QIAamp Viral DNA Mini Kit, HBV DNA s region was obtained by nested PCR and sequencing.

RESULTS12 of the 140 children were HBV DNA detected were positive and the infectious rate was 8.6% . No mutant of the 12 HBV DNA in "a" determinant.

CONCLUSIONTo evaluate the effection of the prevention of HBV mother-to-child transmission, the standard method should be established. The detection of HBV DNA should be included in the future.

Child ; Child, Preschool ; DNA, Viral ; blood ; genetics ; Female ; Hepatitis B ; blood ; prevention & control ; transmission ; Hepatitis B Surface Antigens ; blood ; genetics ; Hepatitis B virus ; genetics ; Humans ; Infectious Disease Transmission, Vertical ; prevention & control ; Male ; Mutation

Objective:To prepare compound fluconazole gel and to establish a method for its quality control. Methods: Flucon-azole and baicalin were the main ingredients in the gel, and their contents were determined by HPLC. Moreover, the drug release in vitro, irritation and stability of the gel were tested. Results:The fluconazole and baicalin had a good linear relationship within the range of 12. 5-150. 0 μg·ml-1 and 25. 0-175. 0 μg·ml-1, respectively. The average recovery rate was 99. 45%(RSD=0. 40%, n=9) and 99. 31%(RSD=0. 31%, n=9), respectively. The in vitro drug release of the gel was accordance with the first order release e-quation, and the cumulative release rate of fluconazole and baicalin in 12 h was 87. 6% and 80. 03%, respectively. The gel was stable without irritation. Conclusion:The formula, preparation technology and stability of compound fluconazole gel are promising and the quality standard is controllable.

Objective To explore whether CD44 +/CD24 + expressing cervical cancer cells are resistant to X-ray irradiation and investigate the underlying mechanism.Methods Cervical cancer cell line (Siha) was cultured in vitro and the CD44 +/CD24 + expressing cells were sorted with a flow cytometer.The cells were irradiated with 8,16 and 30 Gy of 6 MV X-rays.Colony formation test was used to evaluate the radiosensitivity of CD44 +/CD24 + expressing cervical cancer cells.Cell morphology was observed by electronmicroscopy,cell apoptosis was analyzed with a flow cytometer and also verified with a DNA ladder assay.Gene expression was determined by RT-PCR.Results After radiation,the ratio of CD44 +/CD24 + cells significantly increased.Compared to Siha cells,the radiosensitivity of CD44 +/ CD24 + cells decreased (t =93.99-400.45,P ＜0.05),and the expressions of bcl-2,survivin and Oct4 mRNA increased in CD44 +/CD24 + cells (t =221.35,941.65,82.27,P ＜0.01).Both apoptotic body and specific DNA ladder pattern were observed in cells but not in the CD44 +/CD24 + Sihacells which had no obvious morphological changes of apoptosis.Conclusions The CD44 +/CD24 + expressing cervical cancer cells are resistant to X-rays due to expression of anti-apoptosis factors.

Background and purpose:One of the reasons why cancer cells are resistant to chemotherapy is the existence of cancer stem cells. The purpose of this study was to investigate the chemoresistance of CD44+/CD24+ Siha cells to cisplatin and its mechanisms.Methods:Siha cells were cultivatedin vitro. The CD44+/CD24+ Siha cells were sorted out by fluorescence activated cell sorter (FACS) andin vitro proliferation was detected by MTT assay after treatment with the different concentrations of cisplatin. The cell apoptosis rate was detected by flow cytometry after 10 μg/mL cisplatin acted on CD44+/CD24+ Siha cells for 24, 48 and 72 h. The relative mRNA and protein expressions of Bcl-2, Oct-4 and ABCG2 were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. Results:The survival rates of CD44+/CD24+ Siha cells treated with different concentrations of cisplatin (0.1, 1, 5, 10, 15 and 20 μg/mL) were higher than those of their parental Siha cells [(88.42±1.51)%vs (92.87±1.5)%, (79.94±1.05)%vs (84.72±1.09)%, (69.78±0.81)%vs (75.13±2.86)%, (58.97±0.70)%vs (65.79±2.71)%, (49.60±0.88)%vs (52.10±0.52)%, (45.13±0.69)%vs (48.84±1.02)%,P<0.05]. Compared with their parental Siha cells, the apoptosis rates of CD44+/CD24+ Siha cells were lower after 10 μg/mL of cisplatin acting on them for 24, 48 and 72 h, respectively [(3.05±0.16)%vs (5.17±0.27)%, (17.94±2.02)%vs (32.60±4.28)% and (40.14±3.01)%vs (56.62±5.32)%,P<0.05]. The results from both qRT-PCR and Western blot indicated that Oct-4, ABCG2 and Bcl-2 were highly expressed on CD44+/CD24+ Siha cells. A significant difference was found in Oct-4, ABCG2 and Bcl-2 expression between CD44+/CD24+Siha cells and their parental cells (P=0.015<0.05).Conclusion:CD44+/CD24+ Siha cells could be resistant to apoptosis induced by cisplatin and expressed high levels of cancer stem cell markers such as Oct-4 and ABCG2. This study lays the basis for useful isolation and further targeted therapy of cervical cancer stem cells.

Based on microfabrication technology and electrochemical modification method, a micro electrochemical sensor for nitrate ( NO-3 ) determination was developed. A micro sensor chip with working electrode and counter electrode was used as the signal convertor of the sensor. The area of the micro working_electrode was only 1 mm2 . As an electrocatalysis sensitive material, copper was electrodeposited onto the working electrode by square_wave pulse current electrodeposition method. The morphologies and components of freshly deposited materials were examined by scanning electron microscopy ( SEM ) and X_ray diffraction ( XRD) to explore key factors that affected the electrocatalytic ability of the deposited copper layer for reducing nitrate ions. The experimental results revealed that under the optimal conditions, the deposited copper layer was macroporous and had a larger effective surface area that could serve as a more effective electrocatalyst in facilitating nitrate reduction. Electrochemical response of the macroporous copper layer was characterized by linear sweep voltammetry in acidic supporting electrolytes ( pH=2 ) . The electroanalytical results showed that the modified microsensor had marked sensitivity for standard nitrate samples within the concentration range from 12. 5 to 3000 μmol/L (in the range of 12. 5-200 μmol/L yielded straight line:y1=-0. 1422x-10. 326, R12=0. 9976, while in the range of 200-3000 μmol/L yielded straight line: y2=-0. 0984x-22. 144, R22=0. 9927) with a detection limit of 2 μmol/L (S/N=3). The developed electrochemical microsensor was also employed for nitrate determination in water samples collected from lakes and rivers near the city of Beijing. The results were in good agreement with the data given by qualified water quality detection institute, with the deviations from 3 . 9% to 15 . 4%.

Objective To investigate a dose response curve based on a genetic workstation with automatic analysis system of dicentric chromosome assay (DCA) for establishing a high speed dose estimation method.Methods Peripheral blood from three healthy volunteers was irradiated in vitro using 60Co γ-rays,and then lymphocytes were cultured and fixed on slides using the standard protocol for DCA.Dicentric chromosome in metaphase cells was analyzed automatically with the genetic workstation and confirmed manually,and the dose response curve of automated dicentric chromosome was fitted.Dicentric chromosome of another peripheral blood sample irradiated with different doses was manually analyzed to verify the accuracy of the above automated DCA.Results The yield of automated DCA was well fitted by an equation Y =0.018 06D2 + 0.012 79D + 0.000 489 1 with a correlation coefficient R2 =0.961.The biological dose of radiation could be accurately estimated by this dose response curve within a few minutes.Conclusions We had successfully established a new dosimetry method by analyzing dicentric chromesome automatically,which can save a lot of manual analysis time and hence has important significance for emergency rescue in nuclear accidents.

Cinobufacino injection is a significant anti-tumor medicine for the treatment of various tumors in clinic, which was made from water extraction of the skin of Bufo bufo gargarizans. In present paper, HPLC-DAD-FT-ICR-MS method was used to identify the major bufadienolides in cinobufacino for the first time. Solid-phase extraction with dichloromethane and silica was used to enrich the total bufadienolides in cinobufacino. Based on the UV and high resolution MS/MS data, 33 bufadienolides were analyzed and characterized. Among them, eight compounds were identified by comparing with standard references unambiguously. This study elucidated the major bufadienolides in cinobufacino, which provided material foundation of cinobufacino and will be benefit for the further pharmacological research.
Amphibian Venoms ; chemistry ; Animals ; Bufanolides ; analysis ; chemistry ; Bufo bufo ; Chromatography, High Pressure Liquid ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

To obtain chemical constituent information of rat plasma after oral administration of Huang-Lian-Jie-Du Decoction (HLJDD), a LC-FT-ICR-MS method has been established, and both positive and negative ions scan modes were include in the analysis. By comparing their retention time, high resolution mass data of HLJDD extracts, blank plasma and dosed plasma, 38 constituents, including 22 prototype compounds and 16 metabolites, were detected in rat plasma after oral administration of HLJDD. In the 22 prototype compounds, 16 constituents were determined unambiguously by comparing with references. In the analysis of metabolites, phase II reactions like glucuronidation and sulfation were the major biotransformation pathways of HLJDD. M11 was observed as the only phase I metabolite in present experiment. The results will be beneficial for the further pharmacokinetics and pharmacological evaluations of HLJDD.
Administration, Oral ; Alkaloids ; blood ; Animals ; Biotransformation ; Chromatography, High Pressure Liquid ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; metabolism ; Flavonoids ; blood ; Iridoids ; blood ; Male ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

Objective To identify the species of cultured cells and to detect inter-species cross contamination by polymerase chain reaction (PCR).Methods From references and NCBI database,we outsourced 32 pairs of species-specific primers targeted to genomic DNA of ten common species of cultured cells.Then we screened for optional primers with high specificity and high sensitivity.PCR amplification with these primers,the genomic DNAs isolated from the tested cell line and agarose gel electrophoresis of the PCR products were done.Mixed DNA of 10 species was used as positive-control template,and water as negative-control template.Results Ten pairs of species-specific and highly sensitive primers were selected.By PCR amplification with these primers and agarose gel electrophoresis,we may easily identify the origin of cell lines and find whether the tested cell lines are contaminated by cells of other species.Conclusion This PCR assay provides a simple,rapid,sensitive,and cost-effective method to identify cell species and detect interspecies cross-contamination.

Objective To compare the applicability of different occupational health risk assessment methods in small furniture manufacturing industry. Methods American EPA inhalation risk model, Singapore semi-quantitative risk assessment model of occupational exposure to chemical substances and Australian Occupational Health and Safety Risk Assessment model, were used to assess the occupational health risk in a small furniture manufacturing industry from a city of Zhejiang Province. Results The results of American EPA model showed that the workers who exposed to benzene and formaldehyde had low risk of carcinogens, and who exposed to benzene and xylene had very high risk of non-carcinogens. According to Singapore semi-quantitative risk assessment model, there were low and medium health risk caused by toluene and xylene, and high risk caused by wood dust in preparation and polishing jobs. Similar to the results of other models, Australia qualitative risk assessment model showed that there were medium health risk caused by toluene and xylene, and high risk caused by benzene, wood dust and noise. All of the three methods could found the key risk control point in furniture manufacturing industry. The risk ratios of the three methods were higher than the toxic work classification ratio (P<0.01), and the risk ratio of EPA model were higher than the results of Singapore model and Australia model (P<0.05) . Conclusion All of the three methods can be applied to assess the occupational health risk in furniture manufacturing industry, and the combined application of multiple risk assessment methods can be used as one of the risk assessment strategies.