Ammonia ; blood ; Humans ; Liver Failure ; blood ; pathology ; therapy ; Liver, Artificial ; Urea ; blood

Objective To evaluate effects of chronic arsenic exposure and arsenic exposure time on oxidative DNA lesions in humans. Methods A cross-sectional study was conducted in 108 subjects exposed to high concentrations of arsenic in drinking water and 75 control subjects. A cohort study was conducted in 64 subjects exposed to high levels of arsenic in drinking water for 7 or 9 years. Urinary 8-oxo-7,8-dihydredeoxygnanine(8-OHdG) levels were analyzed by the enzyme-linked immunosorbent assay kit(ELISA). Urinary arsenic concentration was detected with hydride generation atomic absorption spectroscopy. Results In the cross-sectional study, the median of urinary arsenic concentration was 484.17 mg/kg Cr for the arsenic-exposed group, and 13.80 mg/kg Cr for the control group, and the difference between the two groups was statistically significant (t=32.57, P<0.01). The median of urinary 8-OHdG levels was 16.60 and 21.88 mg/kg Cr for arsenic-exposed children and adults respectively, much higher than control children(10.50 mg/kg Cr) and adults (9.11 mg/kg Cr), and the difference was statistically significant (t=5.049, 6913, all P<0.01). Urinary 8-OHdG levels were signifieandy lower for children than adults in the exposed group(t=-1.997, P<0.05). In the cohort study, the median of urinary arsenic concentration was 461.3 mg/kg Cr for the 7-year-exposed subjects and 422.90 mg/kg Cr for the 9-year-expesed subjects, and no significant difference was observed(t=-0.250, P 0.05). The median of urinary 8- OHdG levels for 9-year-exposed children and adults were 23.46 and 24.30 mg/kg Cr respectively, significantly increased compared with those of 7-year-exposed(14.29 and 18.38 mg/kg Cr), and the difference had statical signhqcanees (t= -2.949,-3.055, all P<0.01). Conclusions Chronic arsenic exposure can lead to oxidative DNA lesions in humans. The arsenic-induced DNA lesions may aggravate with the exposure time in a certain period.

OBJECTIVETo summarize the experience of a single treatment using molecular adsorbent recirculating system (MARS) in patients with acute-on-chronic liver failure.

METHODSTwenty-five eases treated by MARS-artificial liver were followed up and reviewed.

RESULTSThe levels of serum total bilirubin, non-conjugated bilirubin and blood ammonia were significantly decreased from (618.51 200.68) mmol/L to (390.81 146.02) mmol/L (t=2.729, P<0.01), (490.03 163.39) mmol/L to (303.28 113.06) mmol/L (t =2.516, P<0.01), and (152.44 82.62)mmol/L to (84.80 13.30)mmol/L (t=2.174, P<0.05), respectively. Prothrombin activity was significantly increased from 70.55% 32.39% to 93.63% 14.20% (t=1.728, P<0.05) in patients during a single 6 h treatment with MARS. No difference was presented in the markers of liver zymogram, serum protein, kidney function, electrolyte, blood routine and blood gas analysis before and after the MARS. Thirteen of 17 patients have been cured or improved, 4 died, and the survival rate was 76.5%.

CONCLUSIONSMARS is a safe and an effective treatment for patients with liver failure.

Adult ; Aged ; Ammonia ; blood ; Bilirubin ; blood ; Chronic Disease ; Female ; Follow-Up Studies ; Humans ; Kidney Function Tests ; Liver Failure, Acute ; blood ; therapy ; Liver Function Tests ; Liver, Artificial ; adverse effects ; Male ; Middle Aged

AIMTo analyze the chemical components in Danggui (the roots of Angelica sinensis (Oliv.) Diel).

METHODSHPLC-MS/MS was used to identify the main components in Danggui. Furthermore, the MS fragmentation regularity of the phthalides was proposed. The mobile phase of HPLC consisted of 0.5% acetic acid in water and 0.5% acetic acid in acetonitrile, analytical column was Hypersil ODS2 (250 mm x 4.6 mm, 5 microm), flow rate 1.0 mL x min(-1), injected volume 2 microL. The ionization source was ESI in positive ion mode.

RESULTSFerulic acid, nine known phthalides and one unknown phthalide derivative were tentatively identified in chromatograms based on their MS data and the comparison of their UV spectra with those published in the literatures.

CONCLUSIONThe structural information of phthalides was obtained via HPLC-MS/MS, which provides an accurate and fast method to identify the phthalides and provides more scientific information for quality control of Danggui.

4-Butyrolactone ; analogs & derivatives ; analysis ; chemistry ; Angelica sinensis ; chemistry ; Benzofurans ; analysis ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Coumaric Acids ; analysis ; chemistry ; Molecular Structure ; Phthalic Anhydrides ; analysis ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Spectrometry, Mass, Electrospray Ionization ; methods

Danshen,an efficacious agent for cardiovascular diseases,has been found to play an essential role in kidney injury.In the present study,the effect of Danshen on cisplatin-induced renal dysfunction was investigated in a mouse model.Danshen was administered to mice at a dose of 3 g/kg 4 days before and 3 days after cisplatin treatment.A single intraperitoneal injection of 20 mg/kg cisplatin was used to induce nephrotoxicity.The mice were sacrificed 72 h after cisplatin intoxication.Biochemical parameters including serum creatinine and blood urea nitrogen were analyzed.Histopathological changes of kidney tissues were detected using HE staining.Antioxidant enzymes (GSH-Px and SOD) and peroxidative product (MDA) were detected.Protein expressions of Nrf2 and its target genes including HO-1 and NQO1 were measured by Western blotting.The results showed that pretreatment with Danshen significantly reduced serum creatinine and blood urea nitrogen in the cisplatin-treated mice.Histopathological examination showed that Danshen mitigated the renal damage induced by cisplatin.Moreover,Danshen restored the activities of antioxidant enzymes (GSH-Px and SOD) and normalized the MDA contents in renal tissues.Western blotting revealed that Danshen enhanced the expressions of Nrf2 and its target genes in cisplatin-exposed mice.It was suggested that Danshen protects against the cisplatin-induced renal impairment in the mice,which is potentially associated with the upregulation of Nrf2-mediated signaling pathway.

Objective To investigate the problems during using, standard and quality management of the POCT glucose meters in hospital, to analyze the solutions, and to provide reference data for improving the test level of POCT in hospital.Methods The amount, brand and application of portable glucose meters in the hospital were obtained by 3 rounds of surveillance from May to July in 2013.All of those glucose meters were taken part in external quality assessment of Clinical Laboratory Center of National Health and Family Planning Commission.The test results of those glucose meters were compared with that of automatic biochemical analyzer, the comparison results were then analyzed.Results The POCT glucose meters possessed 5 brands in our hospital, and the amount and type of glucose meters in some clinical departments were often changed.When 4 brands which were detected as quality control samples by ministry of health, the accuracy of detection results of 3 concentration of brand Ⅲ were substandard, the CV% of two levels were 11.9%and 10.1%respectively, the remaining 3 brands were in line with the requirements.The qualified percentages of 3 times of comparison were 85.0%, 92.0%and 97.4%.Conclusions The hospital should select the brand of portable glucose meters reasonably, correct use of glucose meters, and it is very necessary to build indoor and interstitial quality evaluation system, at the same time, suggesting the hospital to establish the POCT quality management team, to carry out the instrument comparison periodically, so to guarantee the accurate, reliable results of POCT in hospital.

Objective To evaluate the impact of Shenfushu granules on cisplatin-induced renal dysfunction and explore the possible mechanisms.Methods Male institute of cancer research(ICR) mice were randomly divided into 4 groups:control group (0.9％ NaC1),Shenfushu group (Shenfushu granules 1 g · kg-1),cisplatin group(0.9％ NaC1 for 9 d and a single dose of cisplatin 20 mg · kg-1 was given by intraperitoneal injection on 7th day),Shenfushu + cisplatin group (Shenfushu granules 1 g · kg-1 gavage for 9 d and a single dose of cisplatin 20 mg · kg-1 was given by intraperitoneal injection on 7th day).Animals were euthanized 72 h following cisplatin dosing.Biochemical parametem including serum creatinine and blood urea nitrogen levels were analyzed by automatic biochemical analyzer.Hematoxylin-eosin (HE) staining was performed inhistopathological examination.The peroxidative product malondialdehyde (MDA) and antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were detected according to kits.Protein expressions of nuclear factor-E2-related factor 2 (Nrf2),hemeoxygenase 1 (HO-1),quinine oxidoreductase 1 (NQO1),multidrug resistance protein 2 (MRP2) and multidrug and toxic compound extrusion protein 1 (MATE1) were measured in western blot analysis.Platinum contents in kidney were determined with inductively coupled plasma mass spectrometry (ICP-MS) method.Results The serum creatinine in cisplatin group and Shenfushu + cisplatin group were (12.14 ± 4.64),(7.60 ± 3.49) μmol · L-1;blood urea nitrogen were (34.12 ±9.48),(15.92 ±4.33) mmol · L-1;GSH-Px were (78.26 ±7.02),(95.87 ± 14.08) U· mg-1 protein;SOD were (8.85 ±3.99),(12.68 ±3.62) U · mg-1 protein,MDA were (6.96 ±2.25),(4.33 ± 1.50) nmol · mg-1 protein;platinum contents were (12.56 ± 1.30),(9.67 ± 0.97) ng · mg-1 tissue.Western blotting analysis revealed that Shenfushu granules enhanced expressions of Nrf2-mediated antioxidant pathway and efflux transporter in cisplatin-exposed animals.Conclusion Shenfushu granules exhibited renal protective effects on cisplatin-induced renal impairment,possibly associated with its modulating on Nrf2-mediated antioxidant pathway and efflux transporters.

Objective To provide technical support for market supervision of TCM decoction pieces in Beijing, Tianjin and Hebei districts; To facilitate the medical treatment of people in the three districts. Methods A comparative study and analysis on processing procedures of TCM decoction pieces in Beijing, Tianjin and Hebei districts was conducted. Results There were some differences in TCM decoction pieces in Beijing, Tianjin and Hebei districts. The quality of TCM decoction pieces is easy to be contradictory according to different standards. Conclusion It is badly in need of a unified standard for the preparation of TCM decoction pieces.

This study was aimed to clone the gene coding mouse CXC chemokine receptor 4 (CXCR4), to construct the recombinant lentiviral vector carrying enhanced green fluorescence protein (EGFP) and to explore its expression in eukaryotic cells (293FT cells). The full length CXCR4 gene was cloned by RT-PCR using bone marrow cells from C57BL/6 mouse as template and inserted into PCR-Blunt vector. CXCR4 fragment was generated by digestion with restriction endonuclease and subcloned into a lentiviral vector to generate recombinant lentiviral vector LV-IRES-EGFP-CXCR4, which was co-transfected into 293FT cells together with envelope plasmid and packaging plasmid by lipofectamine 2000. Viruses were gathered and concentrated using ultracentrifuge, and then transfected into 293FT cells. Expression of EGFP was detected by fluorescent microscopy and flow cytometry (FCM). And the expression of CXCR4 protein was detected by Western blot. The results demonstrated that mouse CXCR4 gene was cloned and the lentiviral vector was successfully constructed. The lentiviral particles were correctly packaged, and the virus titers were above 10(8) TU/ml in the supernatant after concentration. Expression of EGFP was detected by fluorescent microscopy in the transfected 293FT cells, and the transfection efficacy > 95% was determined by FCM. Expression of CXCR4 protein detected by FCM and Western blot was significantly higher than that in control group. It is concluded that the CXCR4 gene along with the gene coding EGFP is successfully inserted into a lentiviral vector to construct a recombinant lentiviral vector, which can be expressed in eukaryotic cells.
Animals ; Cell Line ; Gene Expression ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Lentivirus ; genetics ; Mice ; Mice, Inbred C57BL ; Plasmids ; Receptors, CXCR4 ; genetics ; Transfection

OBJECTIVETo construct TIMP-1 siRNA eukaryotic expression vectors and evaluate their effect on TIMP-1 mRNA expression in hepatic stellate cells.

METHODSThe combinant lone DNA with cutting sites of BamH I and Xho I enzyme according to the sequences of 447-465, 552-540 TIMP-1 of rats and nonspecific sequence were selected and cloned to pGEM-T vector and sub-cloned to pRNAT-U6.2. They were then identified by double enzyme digestion analysis and DNA sequencing. Three plasmids were transfected into T6 separately through an oligofectamine package. TIMP-1 mRNA expression was evaluated by RT-PCR.

RESULTSTargeting sequences of TIMP-1 siRNA eukaryotic expression vectors were correct. TIMP-1 mRNA expression was significantly reduced by transfecting them into the T6.

CONCLUSIONWe successfully constructed two TIMP-1 siRNA eukaryotic expression vectors and the transfected cells can significantly suppress the TIMP-1 expression.

Animals ; Cell Line ; Gene Silencing ; Genetic Vectors ; Hepatic Stellate Cells ; Plasmids ; RNA, Small Interfering ; Rats ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; Transfection