Objective To construct the recombinant plasmid pYES6-S and express and purify spike protein of severe acute respiratory syndrome(SANS)coronavirus in Saccharomyces cerevisiae. Methods DNA fragments of SANS coronavirus were obtained by reverse transeription.Four over- lapped fragments of spike protein genes were amplified by polymerase chain reaction(PCR)and ligated into an integral spike protein gene by restriction enzyme digestion.The spike protein gene recombined with pYES6 and cloned into E.coll.The recombinant plasmid pYES6-S was induced and expressed in Saccharomyces cerevisiae(INVScl)by galactose.Results The recombinant plasmid pYES6-S was confirmed that inserted fragment was right in length,direction and base matching by restriction enzyme digestion and sequencing.The purified protein encoded by the whole spike protein gene was about Mr 110?10~3 identified by electrophoresis.Conclusion The whole spike protein gene of SARS coronavirus is cloned into E.coli and the protein is expressed in Saccharomyces cerevisiae successful ly.which can be helpful in SARS vaccine research.

Objectives To investigate the prevalence of animal plague in Horqin Right Front County and provide scientific bases for prevention and treatment of the disease.Methods Plague surveillance was carried out according to the requirements of National Plague Monitoring Programme at 3 monitoring points in Horqin Right Front County in 2012.Squirrel density,the number of small rats,rat fleas,pathogenic and serological surveillance of Yersinia pestis were included in the study.Results A total of 356 dauricus were captured,the average density was 0.88 rats/hm2; female and male ratio was 1 ∶ 1.31 and the female embryo rate was 96.46％.The captured 13 small nocturnal mice belonged to 2 species,which included 10 cricetulus barabensis and 3 cricetulus longicaudatus,and the capture rate of small nocturnal mouse was 4.33％ (13/300).A total of 98 rat fleas were collected from 25 rats; rat flea carrying rate was 100％(25/25) and flea index was 3.92.Eighty rat burrows were detected,and flea index was 0.188.Pathogen test results were negative.Indirect hemagglutination test was used to examine 356 rat sera and 1 serum was positive.Conclusions Detection of positive rat sera has proved the existence of plague epidemic in Horqin Right Front County.Therefore,the work on plague prevention and control has a long way to go.

Objective To evaluate the dyssynchrony of left ventricle in patients with dilated cardiomyopathy by two-dimensional speckle-tracking strain (2DS) and tissue velocity imaging (TVI).Methods Study population consisted of 37 dilated cardiomyopathy patients. High frame rate two-dimensional images were recorded from the left ventricular short-axis views at the levels of mitral annulus,papillary muscles and apex, and the apical four-chamber view, two-chamber view and long-axis of the left ventricle. The time to peak myocardial longitudinal systolic velocity was measured by TVI, 2DS was acquired to measure the time to peak radial strain in the short axis and the time to peak longitudinal strain inthe long axis,left ventricular synchronization index (△T) was defined as the difference of the time to peak value between anterior septum and posterior wall. Results △T measured by 2DS on the long axis and the short axis increased significantly compared with TVI(P<0.01) in the basal segment; 2DS on the short axis had a more significantly increased △T than TVI(P<0.01) in the middle segment. △T measured by 2DS on the short axis significantly increased in the basal and middle segment compared with 2DS on the long axis(P<0.05). Dyssynchrony eases and the detection rate of dyssynchrony measured by 2DS on the long axis were significantly higher than 2DS on the short axis and TVI(P<0.01). Dyssynchrony cases and the detection rate measured by 2DS on the short axis were higher than those measured by TVI, but the difference had no statistical significance. Conclusions The dyssynchrony detection rate measured by longitudinal strain of 2DS is significantly higher than TVI and radial strain.

This paper was aim to optimize the purification technology of Rehmanniae Radix Praeparata extract with macroporous adsorption resin. With the content of manninotriose as index, the absorptive flow and time were investigated, as well as kinds, amount, flow of eluent. D-101 type macroporous adsorption resin was the best choice for the purification of manninotriose. The optimized parameters were as follows: the content of manninotriose at 161.16-53.72 mg x g(-1), absorption time 240 min, eluting solvent of purified water, volume flow at 1.5 BV x h(-1), and eluant volume at 6 BV. D-101 type macroporous adsorption resin could significantly increase the purity of Rehmanniae Radix Praeparata extract with the advantage of high absorption, remove most part of impurity, and the effect of semi-works production was better.
Adsorption ; Chemical Fractionation ; Chromatography, Liquid ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Rehmannia ; chemistry ; Resins, Synthetic ; chemistry

Objective To explore the relationship of expression and the effect of preoperative radiotherapy of endothelin-1 (ET-1) and pyruvate kinase M-2 (PKM2) in rectal carcinoma.Methods Immunohistochemical method was used to detect the expression of ET-1 and PKM2 proteins of rectal cancer tissues in 96 cases.The expressions of ET-1 and PKM2 were analyzed with the effect of preoperative radiotherapy in rectal cancer tissue.Results The high expression of ET-1 protein was 59 cases (61.46％).The high expression of PKM2 proteins was 54 cases (56.25％).The high expressions of ET-1 and PKM2 protein were worsen the effect of tumor regressive grade (TRG) than lower expressions of those after preoperative radiotherapy of rectal cancer tissue (P ＜ 0.05).The protein expression of ET-1 and PKM2 were positively correlated (P =0.006).Conclusions The high expressed ET-1 and PKM2 proteins in rectal cancer are closely related to preoperative radiotherapy resistance.ET-1 and PKM2 proteins are expected to become new targets of radiotherapy sensitivity and radiotherapy sensitization of rectal cancer.

Liver failure is severe liver injury caused by various factors. This article summarizes currently used therapeutic methods for liver failure, and the universally acknowledged therapeutic methods include intensive care and supportive care, etiological treatment, regulation of immune function, prevention and treatment of various complications, supportive treatment with artificial liver, and liver transplantation. It is pointed out that the treatment of liver failure should focus on the multimodality therapy of internal medicine-artificial liver-liver transplantation, and that early diagnosis and treatment should be emphasized in clinical practice. The multimodality therapy should be developed according to the etiology, and complications should be actively prevented and treated. A multidisciplinary team should be established to develop individualized diagnosis and treatment regiments, so as to reduce the mortality rate and improve patients′ clinical outcome.

OBJECTIVETo establish a rapid quantitative analysis method for the content of chlorogenic acid and solid content in the extraction liquid concentration process during the production of Reduning injection by using the near-infrared (NIR) spectroscopy, in order to reflect the concentration state in a real-time manner and really realize the quality control of concentrating process of the extraction and concentration process.

METHODThe samples during the Jinqing extraction liquid concentration process were collected. After the removal of abnormal samples, the spectra pretreatment and the wave band selection, the quantitative calibration model between NIR spectra and chlorogenic acid HPLC analytical value and solid content was established by using PLS algorithm, and unknown samples were predicted.

RESULTThe correlation coefficients between the chlorogenic acid content and the solid content were respectively 0.992 1 and 0.994 0, and the correlation coefficients of the verification model were respectively 0.994 4 and 0.998 4, with the root mean square error of calibration (RMSEC) of 0.814 6 and 2.656 1 and the root mean square error of prediction (RMSEP) of 0.704 6 and 1.876 7 respectively, and the relative standard errors of predictions (RSEP) were 6.01% and 2.93% respectively.

CONCLUSIONThe method is simple, rapid, nondestructive, accurate and reliable, thus could be adopted for the fast monitoring of the chlorogenic acid content and the solid content during the concentration process of Reduning injection extraction liquid.

Ten flavonoids were isolated from the 95% ethanol extract of the seeds of Alpinia galanga Willd. with a combination of various chromatographic techniques, including silica gel, Sephadex LH-20 and preparative HPLC. On the basis of spectroscopic data analysis, they were elucidated as (2R, 3S)-pinobaksin-3-cinnamate (1), (2R, 3R)-pinobaksin-3-cinnamate (2), pinocembrin (3), pinobaksin (4), 3-O-acetylpinobaksin (5), galangin (6), galangin-3-methylether (7), kumatakenin (8), 3-methylkaempferol (9) and (2R, 3R)-3, 5-dihydroxy-7-methoxyflavanone (10). Among them, compound 1 is a new compound, compounds 2, 5 and 10 were isolated from the genus Alpinia for the first time, and others were isolated from this plant for the first time.
Alpinia ; chemistry ; Benzopyrans ; chemistry ; isolation & purification ; Cinnamates ; chemistry ; isolation & purification ; Flavanones ; chemistry ; isolation & purification ; Flavonoids ; chemistry ; isolation & purification ; Kaempferols ; chemistry ; isolation & purification ; Molecular Structure ; Plants, Medicinal ; chemistry ; Seeds ; chemistry

Objective To investigate the methylation situation of let‐7a‐3 promoter in patients with chronic myeloid leukemia (CML) and its clinical significance .Methods The methylation level of let‐7a‐3 promoter in the bone marrow mononuclear cells of 52 CML patients and 25 controls was detected by using the real‐time quantitative methylation‐specific PCR (RQ‐PCR) .Results The non-hypomethylation of let‐7a‐3 promoter was positive in 31 cases(59 .6% ) of 52 CML patients ,while only 1 case(4% ,1/25) in the control group ,the difference between the two groups were statistically significant (P< 0 .01) .The ROC curve analysis showed that the non -hypomethylation of let‐7a‐3 has better specificity for the auxiliary diagnosis of CML .The significantly posi‐tive correlation was found between the non -methylation level of let‐7a‐3 promoter and the BCR/ABL transcription level (r=0 .641 ,P=0 .001) .In contrast ,there was no obvious correlation between the non -methylation level of let‐7a‐3 promoter and the WBC count ,platelet count and hemoglobin levels(P>0 .05) .The non-hypomethylation level of let‐7a‐3 in chronic phase and accel‐erate phase was significantly higher than that in blastic crisis of CML .Conclusion The hypomethylation level of let‐7a‐3 promoter is decreased with disease progression .

Objective To explore role of 64-slices spiral CT in differetiation of acute chest pains.Methods Thirty six patients with acute chest pains were performed 64-slices spiral CT chest angiography.Two-dimensional and three-dimensional reconstruction was performed in all patients by means of multiplanar reconstruction(MPR)(coronal,sgittal oblique),curved planar reformation(CPR), maximum intensity projection(MIP),and volume rendering(VR).All images were blindly reading by two experienced radiologist.DSA were performed at the same time in 16 cases.Results The coronary artery branches,pulmonary artery and aortic artery in all patients were showed clearly,The acute myocardial infarction were showed in 10 cases,The pulmonary artery embolism in 14 cases,The aortic dissection in 6 cases respectively,The Coronary embolism in One case ,pneumothorax In One case The constrictive pericarditis in 1 case respectively.Normal findings in 4 cases.Conclusion 64-slices spiral CT is a useful and noninvasive examination in acute chest pain.