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The change of kirenol, darutigenol and darutoside in Siegesbeckia and its first to ninth processed products were studied, and the ten fingerprints were compared, which provided the experimental basis for the study of Siegesbeckia processing tech- nology. The samples were analysed by HPLC on a SunFire-C18 column (4.6 mm x 150 mm, 5 μm) with gradient elution of acetonitrile (0.1% formic acid)-water (0.1% formic acid) at a flow rate of 1.0 mL x min(-1). Column temperaturewas 30 °C and the detected wavelength was 215, 320 nm. The calibration curves of kirenol, darutigenol and darutoside were linear in the range of 2.180-26.16, 2.900-34.80, and 1.012-6.072 mg x L(-1), respectively, and the average recoveries were 96.4%, 97.2% and 96.3% wit RSD 2.2%, 1.7% and 2.4%. This method was simple, the result was stable and had good repeatability, recovery and precision. The re- sult was the basis of the chemical contents variation in the processing of Siegesbeckia Herbs and further clarifying the effect of the changing.
Asteraceae ; chemistry ; Chemistry, Pharmaceutical ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Temperature

OBJECTIVETo present an established human chordoma cell line for chordoma research.

METHODSThe specimens pathologically identified as chordoma were cultured, using primary tissue culture in vitro. The surviving cells were analyzed by morphology, histochemical stain, cell cycling analysis, karyotype analysis, electron microscopic observation, heterotransplantation and study of invasive capacity in vitro.

RESULTSThe newly established cell line CM-319 has been maintained in continual cultures for over 100 generations in two years. Its morphological observation, histochemical staining properties, electron microscopic observation and heterotransplantation showed the common characteristics of chordoma. The doubling time of cells was about 33 hours. Cell cycle analysis showed: G(1) 55.6%, G(2) 21.9% and S 22.5%, G(2)/G(1) = 1.90. Chromosome analysis showed a hypotriploid feature and the success rate of heterotransplantation was 100%. It is capable of invasion in vitro.

CONCLUSIONCM-319, as a cell line derived from human chordoma cells, may serve for further studies of chordoma.

Animals ; Cell Line, Tumor ; Cell Proliferation ; Chordoma ; genetics ; pathology ; Humans ; Mice ; Mice, Inbred BALB C ; Neoplasm Invasiveness

Objective：To investigate the correct expression of dengue 2 virus 43 strain NS1 gene in transfected BHK-21 cell. Methods：The D2-43 DNA fragment coding for signal peptide plus NS1 protein was cloned between KpnⅠ site and EcoR Ⅰ site of expression plamid pcDNA3.1. The obtained recombinant vector pcDNA-NS1 was transfected into BHK-21 cells with electroporation technique. After selection by G418, resistant clones were screened by RT-PCR and Western blotting test. Results：The RT-PCR results of four in five randomly selected cell clones were positive. Western blotting test showed that NS1 gene could be expressed in BHK-21 cells. Conclusions：NS1 protein was capable of being expressed and appropriately processed in pcDNA-NS1 transfected BHK-21 cells. The present results suggest the feasibility of NS1-based DNA immunization.

ObjectiveTo establish the methodology for identification of clinical isolates of Mycobacterium and to identify the distribution of Mycobacterium species in hospitalized patients with tuberculosis in Heilongjiang province.It would provide the basis for accurate diagnosis of infections with Mycobacterium tuberculosis (MTB) and non-tuberculous Mycobacterium (NTM) as well as for effective therapy.Methods Three hundred and thirty Mycobacterium isolates from 330 tuberculosis patients hospitalized and clinically diagnosed in Harbin Chest Hospital from May 2007 to December 2008 were collected.Genomic DNA from the isolates was extracted after inactivation of Mycobacterium.Molecular identification was carried out using PCR,PCR-restriction fragment length polymorphism (RFLP) and sequencing.ResultsAmong 330 clinical isolates,328 were identified as MTB complex (MTBC),accounting for 99.4％ (328/330); 2 were NTM,accounting for 0.6％ (2/330).Among 328 MTBC isolates,326were MTB,one was Mycobacterium Africanum(M.African) and one was Mycobacterium microti(M,microti),accounting for 99.4％ (326/328),0.3％ (1/328) and 0.3％ (1/328),respectively.It was found that the homology between the two NTM isolates and Mycobacterium avium intracellulare (MAC)was 99％ and 93％,respectively,suggesting that the two NTM isolates were MAC.The homology between the two NTM isolates was 93％.ConclusionsPCR plus PCR-RFLP and sequencing provides an ideal method for precise identification of Mycobacterium species.In the clinically diagnosed tuberculosis patients,the predominant Mycobacterium species is MTB,however M.African,M.microti as well as NTM are also found.

BACKGROUNDCarotid artery intima-media thickness (CIMT) and brachial artery flow-mediated dilation percentage (FMD%) are common parameters used for detecting subclinical atherosclerosis. This study compared subclinical atherosclerosis of the carotid and brachial arteries in rheumatoid arthritis (RA) patients and healthy controls using high resolution ultrasonography. We also investigated their correlation with clinical factors and the association between FMD% and CIMT.

METHODSOne hundred and two RA patients and 46 age-gender matched healthy controls were included in the study. FMD of the brachial artery and CIMT were measured ultrasonographically. Patients with diabetes mellitus, hypertension, renal failure, history of cardiovascular or cerebrovascular disease were excluded. Subjects who were receiving or used high dose steroids were also excluded.

RESULTSThe CIMT was significantly higher in patients than that in the control group ((0.697±0.053) vs. (0.554±0.051) mm, P<0.001), whereas brachial artery FMD% was lower in patients than that in the controls ((5.454±2.653)% vs. (8.477±2.851)%, P<0.001). CIMT was related to age, disease duration, tender and swollen joint score, C-reactive protein, systolic blood pressure and high-density lipoprotein. However, FMD% was only association with systolic blood pressure. There was no significant correlation between CIMT and FMD%.

CONCLUSIONSCompared with the healthy control subjects, RA patients without clinically evident cardiovascular disease had subclinical atherosclerosis in terms of impaired FMD% and increased CIMT. FMD% and CIMT may measure a different stage of subclinical atherosclerosis in RA patients.

Adult ; Arthritis, Rheumatoid ; pathology ; physiopathology ; Atherosclerosis ; pathology ; physiopathology ; Brachial Artery ; physiopathology ; Carotid Intima-Media Thickness ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged

Objective To summarize the experiences of diagnosis and treatment for blunt tracheobronchial injuries ( BTI) . Methods From January 1993 to December 2013, 15 patients were diagnosed with BTI and underwent surgical treatment at our hospital.Mean age of the 15 patients (11 men and 4 women) was 26.4 ±4.5 years.All the patients had a history of trauma, which included crushing injury in 8 cases, deceleration injury in 4, fall injury in 2, and traction-type injury in 1.BTI loca-tion: right main bronchus in eight cases, left main bronchus in four cases, upper trachea in one case, lower trachea with the right main bronchus in one case, and cervico-thoracic trachea with left main bronchus in one case .Thoracic computerized tomo-graphy was performed in 15 patients, which showed pneumothorax, subcutaneous emphysema, pneumomediastinum or falling lung sign of Kumpe.Preoperative fiberoptic bronchoscopy examination was performed in 15 cases, which included bronchial atresia in 9 cases, bronchial transection in 3 cases, laceration of trachea in 2 cases, and tracheal transection in 1 case.An e-lective surgical procedure after BTI was performed in 10 cases, and emergency surgery was performed in 5 cases.Tracheo-bron-chial laceration repair were performed in 2 cases, tracheal end-to-end reanastomosis in 1 case, and bronchial end-to-end re-anastomosis in 12 cases.Results There was no operative death, and one case was complicated with anastomotic stenosis.The average operation time was 205.7 ±41.3 minutes, and the average blood loss was 268.4 ±109 ml.The postoperative hospi-tal stay was 11.6 ±3.7 days on average.Follow-up was completed in 15 patients (mean, 29.3 months), and 15 patients were all symptomatic improvement .Conclusion The most common site of BTI was the right main bronchus near Carina parts .Al-though the diagnosis and treatment are often delayed , our findings indicate that chest CT and endoscopic findings could be used for the diagnosis of BTI.Surgical resection and reconstruction are effective methods to repair BTI successfully even many months after they occur.Often they do not require the resection of pulmonary parenchyma .

At present, the tourism industry has risen into a national strategic pillar industry. The development of the TCM industry has been included in the national strategy. The integration of the two major industries, TCM culture and tourism, is of great significance in promoting the development of the tourism industry and structural transformation. Jiangsu Province has a wealth of TCM cultural tourism resources, with unique industrial development advantages. Based on the analysis of the advantages and current situation of TCM cultural tourism in Jiangsu Province, this article put forward some suggestions for the development of TCM cultural tourism in order to provide a reference for promoting the integration and development of TCM culture and tourism industry.

Objective: To evaluate the effects of tube voltage on image quality in coronary CT angiography (CCTA), the estimated radiationdose, and DNA double-strand breaks (DSBs) in peripheral blood lymphocytes to optimize the use of CCTA in the era of lowradiation doses. Materials and Methods: This study included 240 patients who were divided into 2 groups according to the DNA DSB analysismethods, i.e., immunofluorescence microscopy and flow cytometry. Each group was subdivided into 4 subgroups: thosereceiving CCTA only with different tube voltages of 120, 100, 80, or 70 kVp. Objective and subjective image quality wasevaluated by analysis of variance. Radiation dosages were also recorded and compared. Results: There was no significant difference in demographic characteristics between the 2 groups and 4 subgroups in eachgroup (all p > 0.05). As tube voltage decreased, both image quality and radiation dose decreased gradually and significantly.After CCTA, γ-H2AX foci and mean fluorescence intensity in the 120-, 100-, 80-, and 70-kVp groups increased by 0.14, 0.09,0.07, and 0.06 foci per cell and 21.26, 9.13, 8.10, and 7.13 (all p < 0.05), respectively. The increase in the DNA DSB level inthe 120-kVp group was higher than those in the other 3 groups (all p < 0.05), while there was no significant difference inthe DSBs levels among these latter groups (all p > 0.05). Conclusion The 100-kVp tube voltage may be optimal for CCTA when weighing DNA DSBs against the estimated radiationdose and image quality, with further reductions in tube voltage being unnecessary for CCTA.

OBJECTIVETo investigate the differential expression of RPL6/Taxreb107 between drug-resistant gastric cancer cell line SGC7901/ADR and gastric cancer cell line SGC7901 as well as its correlation with multiple-drug resistance (MDR) in gastric cancer cells.

METHODSTotal RNA was extracted from SGC7901 and SGC77901/ADR, with internal control RT-PCR, Northern blot, gene cloning and expression, construction of eukaryotic expression vector, gene transfection by electroporation. The accumulation and retention of ADR in transiently transfected cell was detected by flow cytometry.

RESULTSThe internal control RT-PCR and Northern blot showed high RPL6/Taxreb107 expression in SGC7901/ADR cell line. Sense and antisense eukaryonic expression vectors demonstrated by double enzyme digestion were successfully transfected into gastric cancer cell line SGC7901 and SGC7901/ADR respectively by electroporation. The accumulation and retention of ADR detected 48 hours after transfection showed that RPL6 gene had shown effect on drug resistance in gastric cancer cell.

CONCLUSIONThe high expression of RPL6/Taxreb107 in drug resistant gastric cancer cell shows its correlation with multiple-drug resistance in gastric cancer.

DNA-Binding Proteins ; metabolism ; Drug Resistance, Multiple ; physiology ; Drug Resistance, Neoplasm ; physiology ; Humans ; Statistics as Topic ; Stomach Neoplasms ; pathology ; Tumor Cells, Cultured