OBJECTIVETo study the chemical constituents in roots of Heracleum rapula.

METHODThe constituents were isolated by column chromatography on silica gel and ODS, and identified by spectroscopic methods.

RESULTEight compounds, xanthotoxol (I), 8-geranyloxypsoralen (II), (+)-marmesin (III), beta-sitosterol (IV), stigmasterol (V), oleanolic acid (VI), ferulic (VII), scopoletin (VIII) were isolated and identified.

CONCLUSIONCompounds IV, V, VI, VII, VIII were isolated from this plant for the first time.

Coumaric Acids ; chemistry ; isolation & purification ; Heracleum ; chemistry ; Oleanolic Acid ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Scopoletin ; chemistry ; isolation & purification ; Sitosterols ; chemistry ; isolation & purification ; Stigmasterol ; chemistry ; isolation & purification

Objective To study the prevalence of mental disorders among the Chinese youths aged 15-34 years,in rural areas and to identify risk factors related to suicide.Methods A consecutive sampling strategy was used for suicidal cases in 16 randomly selected counties in Hunan,Liaoning,and Shandong provinces.Between 2005 and 2008,a total of 392 suicide cases were recruited with 416 community controls at the same age range,selected from the same areas one family member together with one close friend of each suicidal case were interviewed,using the psychological autopsy (PA) method.The same method with structured instruments was performed on the two informants for each control in the same community.SCID was used for the diagnosis of mental disease.Results 48.0％ of the suicides were diagnosed as having at least one mental disorder episode,in comparison with only 3.8％ among the controls.It was found that mental disorder was the most important risk factor for the Chinese young suicide cases in the rural areas.Conclusion As seen in the Western countries,mental disorder had also been the number one correlate on suicidal cases in China,with the difference as other social and psychological factors might have played relatively more important roles in China.

Objective To investigate the antimicrobial resistance of nonfermenting gram-negative bacteria in China in 2012.Methods All the clinical isolates of nonfermenting gram -negative bacteria were collected from 557 tertiary hospitals from 1st January 2012 to 31st December 2012 in China.The antimicrobial susceptibilities of the isolates were de-termined by using various methods such as broth microdilution, disk diffusion, or E-test which according to CLSI 2012 guideline.The sus-ceptibility data were processed with WHONET 5.6 software.Results A total of 198334 nonfermenting gram-negative bacteria susceptibility data were obtained.Among all the isolates, the top 3 isolated nonfermenting gram-negative strains were Pseudomonas aeruginosa ( 80964 strains, 40.8%) , followed by Acinetobacter baumannii(67368 strains, 34.0%) , and Stenotrophomonas maltophilia ( 17520 strains, 8.8%).Against Pseudomonas aeruginosa, polymyxin B, tobramycin, ceftazidime, ciprofloxacin, levofloxacin, cefepime, gentamicin and cefoperazone/sulbactam ( CSL) were the only agents to which >70%of isolates were susceptible.No antibiotics except polymyxin B ( 96.9%) , minocycline ( 61.4%) , amikacin (58.7%), CSL (54.3%) and tobramycin (51.3%) were active more than 50% against Acinetobacter baumannii.Susceptibility rates of carbapenem against Acinetobacter baumannii were around 49%, about 9 percentage points higher than 2011 results.There are some differences among the monitoring results from various districts in China.The suscep-tibilities of Pseudomonas aeruginosa and Acinetobacter baumannii to the tested antimicrobial agents in Central China were generally low, whereas the antimicrobial susceptibilities of Acinetobacter baumannii from North China were genera-lly high.Conclusion The multidrug resistance of nonfermenting gram -negative bacteria, especially Acinetobacter baumannii, has become a worldwide problem.There are some differences among the susceptibilities of isolates from various districts.Polymyxin B has the most effective in vitro antimicrobial activity against Pseudomonas aeruginosa and Acinetobacter baumannii isolates, the emerging resistant isolates should cause enough attention.Further surveillance needed to be strengthened to prevent the further development and spread of resistant bacteria.

OBJECTIVETo establish a nasopharyngeal carcinoma (NPC) cell line CNE1-pLVTHM/BART7 with stable ebv-miR-BART7 overexpression.

METHODSThe recombinant lentivirus pLVTHM/BART7 expression plasmid was packaged into mature lentivirus by 293FT cells and used to infect CNE1 cells. Flow cytometry was employed for sorting the GFP(+) cells. The efficiency of ebv-miR-BART7 overexpression was determined using qRT-PCR.

RESULTSThe recombinant lentivirus plasmid pLVTHM/BART7 was successfully constructed and verified by PCR and sequencing. The expression of ebv-miR-BART7 in CNE1 cells infected with the lentivirus pLVTHM/BART7 was significantly increased as compared with the negative control and the blank control cells.

CONCLUSIONThe recombinant lentivirus vector pLVTHM/BART7 results in high and stable expression of ebv-miR-BART7 in infected CNE1 cells, which provides a useful cell model for further studies of the role of ebv-miR-BART7 in nasopharyngeal carcinoma.

Carcinoma ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; MicroRNAs ; Nasopharyngeal Neoplasms ; genetics ; Plasmids

OBJECTIVETo study the chemical constituents in roots of Heracleum rapula.

METHODThe constituents were isolated by column chromatography on silica gel and ODS, and identified by spectroscopic methods.

RESULTNine compounds, osthol ( I ), bergapten (II), xanthotoxin (III), isopimpinellin (IV), imperatorin (V), isoimperatorin (VI), cnidilin (VII), phellopterin (VIII), rivulobirin A (IX) were isolated and identified.

CONCLUSIONCompounds VI, VII, VIII, IX were isolated from this plant for the first time.

Coumarins ; chemistry ; isolation & purification ; Furocoumarins ; chemistry ; isolation & purification ; Heracleum ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

To explore a rapid and easy method to detect labile iron of pool (LIP) in cells, HL-60 and K562 cells were cultured at a concentration 1 x 10(6)/ml in RPMI 1640 containing 10% heat-inactivated fetal bovine serum. The iron deprivation was induced by adding desferrioxamine (DFO) 10 - 100 micromol/L for 0 - 48 hours. The intracellular LIP was measured by probe calcein-AM. Calcein fluorescence was monitored in 1420 multilabel counter. The results indicated that when HL-60 and K562 cells were incubated with different concentrations of DFO, the calcein fluorescence intensity was higher than that of control group at 12, 24 and 48 hours (P < 0.05). Fluorescence value of representing LIP in DFO groups was lower than that in the control group. In conclusion, DFO can decrease LIP in leukemia cells. The approach used in this study may provide a simple and reliable method for detection of intracellular iron homeostasis.
Cation Transport Proteins ; antagonists & inhibitors ; biosynthesis ; metabolism ; Deferoxamine ; pharmacology ; Fluoresceins ; Fluorescent Dyes ; HL-60 Cells ; Humans ; Iron ; metabolism ; Iron Chelating Agents ; analysis ; metabolism ; Iron-Regulatory Proteins ; metabolism ; K562 Cells

OBJECTIVETo investigate the therapeutic efficacy of interferon (IFN) therapy and risk of long-term administration for chronic hepatitis C (CHC) patients who cannot tolerate the standard treatment.

METHODSForty-six CHC patients who had proven intolerant to standard treatments were treated with low-dose IFN (non-pegylated IFN: 60 to 300MIU QOD, or pegylated IFN: 50 to 90 mug/w) plus ribavirin (RBV; 0.6g to 0.9 g/d) for 72 weeks.

RESULTSForty-three (93.5%) of the patients were able to tolerate the long-term treatment with low-dose IFN plus RBV. Only three patients experienced severe side effects (low white blood cell and platelet counts) that required treatment withdrawal. The virology response rates over treatment time were: rapid virologic response (RVR): 10.9%; early virus response (EVR): 30.4%; 24 week virologic response: 45.7%; and, 48 week virologic response: 47.8%. B-sonographic imaging revealed that three patients experienced improved liver morphology through the treatment course. The patients who achieved RVR, EVR, or 24 weeks virologic response also attained higher 48 week virologic response. The 24 week virologic response had the strongest predictive value of good prognosis.

CONCLUSIONSOur study demonstrated that long-term treatment with low-dose interferon plus ribavirin is effective for patients who are otherwise intolerant to standard treatment. In these patients, low-dose IFN plus RBV can obtain a high virologic response rate at 48 week. Furthermore, the 24 week virologic response is sufficiently predictive of treatment success. As with any treatment regimen, it is important for healthcare workers to monitor the disease status and potential side effects throughout the course of therapy.

Adult ; Antiviral Agents ; administration & dosage ; therapeutic use ; Female ; Hepacivirus ; Hepatitis C, Chronic ; drug therapy ; virology ; Humans ; Interferons ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Treatment Outcome

BpHi006A cDNA is 1943 bp in length, and contains one putative open reading frame that is 795 bp long. The expression of BpHi006A was induced by BPH feeding. BpHi006A protein contains a N-terminal domain and a C-terminal domain of glutathione S-transferase, and therefore, it belongs to the superfamily of glutathione S-transferase. BpHi006A protein exhibited 61% amino acid sequence identity to tetrachloro-p-hydroquinone reductive dehalogenase-related protein of Arabidopsis thaliana. Sequence analysis of these two proteins indicates that they belong to a new group of plant GSTs.
Amino Acid Sequence ; Animals ; Cloning, Molecular ; Genes, Plant ; Glutathione Transferase ; genetics ; Hemiptera ; physiology ; Molecular Sequence Data ; Open Reading Frames ; Oryza ; enzymology ; genetics ; Plant Proteins ; genetics

OBJECTIVE12-0-tetradecanoylphorbol-13 acetate (TPA) plays an important role in precipitating cell differentiation for various tumor cells, especially leukemic cells. Changes of many genes may be involved in this process. The purpose of this study was to observe the relationship between the EGR1mRNA expression and cell differentiation during TPA-induced K562 cell differentiation.

METHODSIncubation of human K562 cells in vitro was applied to cultivate K562 cells. The cells were treated in two different ways. K562 cells of experiment group were treated with TPA and those of control group were treated without TPA. Using morphology (Wright's staining and NSE staining) and flow cytometry (FCM), the investigators observed the differentiation characteristics of K562 cells, cell-cycle and the differentiation antigen expressions of CD33 and CD14 on cell membranes. RT-PCR was carried out to assay EGR1 mRNA expression.

RESULTSAfter treated with TPA for 7 d, the morphology of K562 cells obviously tended to mature differentiation, like monocytes. The differentiation rate of induced K562 cells was up to 95% in experiment group and 4.5% in control group, respectively. Using SPSS software, the above result showed statistical significance (P < 0.01). Using NSE staining, K562 cells showed positive reaction. Some of them were densely stained. The positive rate was up to 86%. More than half of the positive cells could be inhibited by NaF. The inhibiting rate of NaF was up to 58.72%, showing statistical difference when compared with that of control group. FCM analysis showed that most of K562 cells stimulated by TPA underwent G1/S phase cell-cycle arrest. The composing rate of cell-cycle in TPA-treated group showed that (53.7 +/- 1.25)% of cells were at G0 + G1 phase and (44.3 +/- 1.32)% were at S phase (P < 0.05). The level of CD33 expression on cell membranes was mildly decreased from 0.997% to 0.893% (P > 0.05). However, the level of CD14 expression was significantly increased from 0.049% to 0.387% (P < 0.05).

CONCLUSIONK562 cells could express EGR1mRNA during TPA-induced differentiation, which suggested that EGR1mRNA might participate in the process of K562 cells differentiating into monocyte/macrophages, and might play an important role in precipitating and maintaining cell differentiation for leukemic cells.

Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Carcinogens ; pharmacology ; Cell Cycle ; drug effects ; genetics ; Cell Differentiation ; drug effects ; genetics ; Cell Division ; drug effects ; genetics ; Cell Membrane ; chemistry ; drug effects ; DNA-Binding Proteins ; genetics ; Early Growth Response Protein 1 ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Immediate-Early Proteins ; genetics ; K562 Cells ; Lipopolysaccharide Receptors ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Sialic Acid Binding Ig-like Lectin 3 ; Tetradecanoylphorbol Acetate ; pharmacology ; Transcription Factors ; genetics

Peroxisome proliferator-activated receptordelta (PPARdelta), as a downstream target of adenomatous polyposis coli (APC) signaling pathway, has been presumed to play some roles in colorectal carcinogenesis. However, the exact role of PPARdelta in colorectal cancer remains unclear. An HIV-1-based lentivirus packaging system was used for the construction of a lentiviral vector (lentivector) mediating RNA interference against PPARB. The direct sequencing demonstrated that the resulting lentivector containing the short-hairpin RNA expression cassette specifically targeting PPARdelta (sh-PPARdelta) was successfully constructed, and designated as pLVshPPARdelta. The control vector was designated as pLVControl. After the transduction, we observed highly efficient transduction (> 90%) of lentivirus in KM12C cells by fluorescent microscopy and fluorescence-activated cell sorting. Quantitative RT-PCR showed that pLVshPPARdelta lentivirus reduced PPARdelta mRNA expression by about 70.0% in KM12C cells as compared with that of the untreated cells (P < 0.05), while pLVControl had no significant effect on the PPARdelta mRNA level (P > 0.05). Western blot revealed an obvious reduction of PPARdelta protein expression in pLVshPPARdelta treated cells and showed no obvious difference between the control group and the untreated group. The results demonstrated that the lentivector mediating RNAi against PPARdelta was successfully constructed, which could stably knock down the PPARdelta expression in KM12C cells. This study finally provided a new cell model for the study of PPARdelta's function in colorectal cancer.
Cell Line, Tumor ; Colonic Neoplasms ; genetics ; pathology ; Gene Knockdown Techniques ; Genetic Vectors ; genetics ; Humans ; Lentivirus ; genetics ; metabolism ; PPAR delta ; biosynthesis ; genetics ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics