Bupleurum chinense used in the study were cultivated in the experimental fields of Gansu agricultural University for three years. The seeds of B. chinense were collected every 3 days 10 d after the blossom. The result showed that the 1 000-grain fresh weight reached the maximum 43 d after the blossom and then decreased rapidly, at the mature period the fresh weight of seeds were falling to the same level of the dry weight. The dynamic change of the grain dry matter accumulation showed as an S-shape curve, the rapid increase stage was 25-34 d following the flower, and the grain filling was ended 46 d after blossom. Grain filling rate was under the law "fast-slow-fast-slow". And there were two peaks of grain filling rate appeared, after reached the second peak 28 d after the flower the filling rate decreased rapidly and stayed steadily 43 d after flowering. The dehydration rate was also measured at its maximum 43 d following flower. The indexes of seeds all reached the top 52 days following the blossom, when the germination rate reached the peak (34.33%) and water content of seeds was near 10%. The rate of germination and the 1 000-graid weight of seed showed significant positive correlation, while the water content of seeds was found significant negatively correlation with germination percentage. So the best time for harvest should be 52 d after flowering (9 month), the seeds collected at that time showed both high quality and germination rate.
Bupleurum ; chemistry ; growth & development ; Flowers ; growth & development ; Germination ; Seeds ; chemistry ; growth & development ; Water ; analysis

Objective To evaluate the enhancement effect of the thrombus-targeted ultrasound contrast agent on acute thrombus in normal canine femoral vein.Methods Acute thrombi were created in both sides of the femoral vein in ten canine models.Targeted ultrasound contrast agent was given in a dose of(0.06) ml/kg in ten canines via intravenous bolus injection from forelimb of canine,the images of the acute thrombus were taken at 0,2,4,8,12,14 minutes after infusion of ultrasound contrast agent and stored in magneto optical disks.The images were assessed qualitatively by two independent observers and quantitatively using acoustic densitometry(AD) to determine the contrast enhancement effect of acute thrombus in normal canine femoral vein.Results After infusion of targeted ultrasound contrast agent,the thrombus was enhanced considerably and easy to be detected.AD analysis demonstrated that peak indencity(PI) and area under curve(AUC) increased with the changes of time,reaching the peak at 8th minutes after infusion of targeted ultrasound contrast agent,then decreased.PI and AUC increased at the 8th minute after infusion of targeted ultrasound contrast agent than before [((333.21)?(38.56))dB vs((168.18)?(28.18))dB,((884.40)?(94.62))dB vs((439.65)?(98.54))dB,respectively,P

Objective: To chine and express the recombinant human endothelial monocyte-activating polypeptide-Ⅱ(EMAP-Ⅱ)and identify its anti-tumor biological activities.Methods: EMAP-Ⅱ_(147-312)was expressed by the expression vector pMAL-p2x and E.coli BL-21 and the product was purified.The production of tissue factor(TF)in human umbili- cal vein endothelial cell ECV-304 mediated by the recombinant EMAP-Ⅱwas determined by chemiluminescence sub- strate.The promoting effect of recombinant EMAP-Ⅱon TNF?-induced ECV-304 cell.Apoptosis was determined by flow cytometry.Its inhibitory effect on human pancreaic cancer cell SW1990 proliferation was determined by MTT method. Results:DNA sequencing verified that EMAP-Ⅱwas correctly cloned.The molecular mass of the protein identified by SDS-PAGE was consistent with the theoretic value.The productivity of recombinant EMAP-Ⅱwas 500?g per 1 g bacteria (wet mass).The purified product induced expression of tissue factor(TF)in ECV-304 cells;it also enhanced the sensi- tivity of ECV-304 cells to the apoptotic effect of TNF?([16.6?2.5]% vs[25.6?2.3]%,P

Objective:To express Tumstatin_(183-230)-TRAIL fusion protein and to observe its biological functions.Methods: SOE-ing PCR was employed to amplify the recombinant sequence of Tumstatin_(183-230)and TNF-related apoptosis-inducing ligand (TRAIL_(114-281)).An expression vector pMAL-Tu-T was constructed by inserting Tu-T sequence into pMAL-c_2;the vector was used to transfect E.coli BL21(DE3)and expression of MBP-Tu-T fusion protein was induced by IPTG.Amylose Resin columns were employed to purify the fusion protein.The biological functions of MBP-Tu-T protein was examined by inhibitory test of endothelial cell proliferation,standard tumor cell cytotoxic assay,in vitro tube formation inhibition,and electron microscopic observation(apoptosis).Results:The expression rate of MBP-Tu-T fusion protein in E.coli was about 20%. Purified recombinant protein obviously inhibited endothelial cell proliferation(IC_(50)12.5?g/ml),induced apoptosis of pancreatic cancer cells,and inhibited tube formation.Conclusion:Constructed MBP-Tu-T fusion protein is bifunctional,which lays a solid foundation for further investigation of antitumor effect of Tumstatin_(183-230)-TRAIL in vivo.

OBJECTIVETo provide the basis for clinical acquired immunodeficiency syndrome (AIDS) surveillance and to avoid cross infection in hospital, we study the infection status of AIDS in Shandong province.

METHODSThe fourth-generated Akzo's ELISA kit and the fourth generated Immunoluminometric detection reagent were used for HIV antibody screening for 399 303 cases of both inpatients and outpatients from Jan. 2003 to Dec. 2011. Beijing WanTai ELISA kit and Se-marked rapid detection reagent were used for re-detection, and the positive samples were sent to the local CDC for confirming test by Western Blot.

RESULTSThe HIV-1 antibody detection results of 129 (0. 3230 per thousand) patients were confirmed to be positive, including 54 (0. 1352 per thousand) cases of outpatients and 75 (0. 1878 per thousand) cases of inpatients. HIV infection rates in outpatients from 2003 to 2011 were 0.050 per thousand, 0.030 per thousand, 0.111 per thousand, 0.120 per thousand, 0.124 per thousand, 0.113 per thousand, 0.148 per thousand, 0.201 per thousand, 0.2152 per thousand; and that in inpatients were 0. 150 per thousand, 0.089 per thousand, 0.138 per thousand, 0. 144 per thousand, 0. 104 per thousand, 0. 132 per thousand, 0. 197 per thousand, 0. 329 per thousand, 0. 313 per thousand respectively. Among these inpatients, there were 61 cases of medical patients and 14 cases of surgical patients, and most were youths and farmers.

CONCLUSIONSHIV infection rate was increasing year by year. Most inpatients whose HIV-1 antibody was positive were in the phase of AIDS. Therefore, it's very necessary to execute routine testing for inpatients and outpatients who need special examination for early diagnosis of HIV infection

Acquired Immunodeficiency Syndrome ; epidemiology ; immunology ; virology ; Adolescent ; Adult ; Aged ; China ; epidemiology ; Enzyme-Linked Immunosorbent Assay ; Female ; HIV Antibodies ; analysis ; HIV Infections ; epidemiology ; immunology ; virology ; Humans ; Male ; Middle Aged ; Young Adult

Objective To explore the relationship between immune and pathogenesis of amyotrophic lateral sclerosis (ALS) in the investigation of neurofilaments phosphorylation and ultrastructure features in spinal cord ventral horn motor neuron injury mediated by immune.Methods Using transmission electron microscope,we studied the uhrastructure features of abnormal accumulations of neurofilaments (NF) in motoneuron of the spinal cord ventral horn,and immunohistochemically investigated neurofilaments phosphorylation.Results Electron microscope found that there was abnormal accumulation of interwoven NFs in motor neuronal perikarya and proximal axons.Immunohistochemical study revealed that the SMI-32 immunoreactive positive neurons (12.00?1.05),compared with control (18.00?1.83),were reduced (P

OBJECTIVETo compare the efficacy and toxicity of concurrent chemoradiotherapy followed by consolidation chemotherapy (CCRT-CT) and sequential chemoradiotherapy (SCRT) in the treatment of stage III non-small cell lung cancer.

METHODSFrom February, 2007 to June, 2010, 93 patients with unresectable stage III non-small cell lung cancer were treated with SCRT or CCRT-CT. SCRT group (50 cases) received radiotherapy after 2-6 cycles of chemotherapy (median 2 cycles) followed by 0-4 cycles (median 2 cycles) of chemotherapy. CCRT-CT group (43 cases) received 2 cycles of chemotherapy every 3 weeks with concurrent radiotherapy followed by 2-4 cycles (median 2 cycles) of chemotherapy with the same drugs. The chemotherapy consisted of cisplatin plus gemcitabine, docetaxel or vinorelbine. Radiotherapy was administered using two-dimensional conformal irradiation (36-40 Gy/18-20f) followed by three-dimensional conformal boost to 56-70 Gy/28-35f (median DT64Gy) or using three-dimensional conformal irradiation 50-74 Gy/25-37f (median DT62Gy).

RESULTSThe response rates were 76.7% and 54.0% in CCRT-CT and SCRT group, respectively (P<0.05). The median progression-free time in the two groups was 16.0 and 10.0 months, with the overall survival time of 18.0 and 12.5 months, respectively. The 1-, 2- and 3-year overall survival rates were 83.7%, 48.8% and 20.9% in CCRT-CT group and 52.0%, 20.0%, and 2.0% in SCRT group, respectively (P<0.05). CCRT-CT group showed a significantly lower rate of distant metastasis than SCRT group (P<0.05), but the local recurrence rate was similar between the two groups. The main side effects included radiation pneumonitis, radiation esophagitis, nausea/vomiting and anemia/leucopenia/thrombocytopenia. CCRT-CT group had a significantly higher rate of III-IV grade nausea/vomiting and anemia/leucopenia/thrombocytopenia than SCRT group.

CONCLUSIONCompared to SCRT, CCRT-CT can improve the response rate, progression free survival and overall survival and decrease the rate of distant metastasis, but is associated with a higher toxicity.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; radiotherapy ; therapy ; Chemoradiotherapy ; methods ; Combined Modality Therapy ; Consolidation Chemotherapy ; methods ; Female ; Humans ; Lung Neoplasms ; radiotherapy ; therapy ; Male ; Middle Aged ; Neoplasm Staging ; Survival Analysis

OBJECTIVETo develop a new nonisotopic detection method of enzyme-amplified time-resolved fluorescence (EATRF) or enzyme-amplified lanthanide luminescence (EALL) for nucleic acid hybridization assays, which can be applied extensively in clinical diagnosis.

METHODSThe method combines the high affinity of biotin-streptavidin system, amplification of enzyme, and inherent advantage of lanthanide chelate with the background elimination of time-resolved fluorescence detection. The conversion of 5-fluorosalicyl phosphate to 5-fluorosalicylic acid (5-FSA) by alkaline phosphatase. The salicylic acid product forms a luminescent ternary chelate with Tb3+ and EDTA.

RESULTSThe dynamic range of the standard curve of EATRFA for nucleic acid hybridization assay was very wide, the range was more than third order of magnitude. The detection sensitivity was about 10 pg of target sequence. When the known target sequence was 20, 10 and 2 ng, the ratio of measured amount to known amount was 110%, 90% and 115% respectively. The main experimental conditions, for example, the irradiating time of ultraviolet rays, the concentrations of biotinylated probe, AP-SA, 5-FSAP and Tb-EDTA and the methods of washing in the related steps, have been optimized. A new stable technology of fluorescence has been developted.

CONCLUSIONSEATRF detection for nucleic acid hybridization assays is a new sensitive simple method, which has a great prospect.

Alkaline Phosphatase ; analysis ; Blotting, Southern ; DNA ; genetics ; Fluoroimmunoassay ; methods ; Luminescent Measurements ; Metals, Rare Earth ; Nucleic Acid Hybridization ; methods ; Spectrometry, Fluorescence ; Substrate Specificity

OBJECTIVETo investigate the economic burden of patients with acute and chronic hepatitis B, cirrhosis and liver cancer caused by hepatitis B virus (HBV).

METHODSCluster sampling was used on cases consecutively collected during the study period. Questionnaire survey was conducted and information on the expenses during hospitalization was collected from the hospital records and through interviewing those patients.

RESULTSYearly costs related to patients with acute hepatitis B, severe hepatitis B, chronic hepatitis B, cirrhosis, hepatocellular carcinoma were 66.7, 138.1, 127.4, 151.7 and 377.2 thousand Yuan, respectively.

RESULTSfrom multiple linear regression model showed that the type of medical insurance scheme, annual days of hospitalization, classifications of HBV-related diseases and personal income were major influencing factors on the cost.

CONCLUSIONHBV infection caused considerable burden to families and the society, indicating that HBV infection control programs would bring huge potential benefits. The reform of insurance scheme should be administrated to promote social fairness.

Adult ; Carcinoma, Hepatocellular ; economics ; China ; Cost of Illness ; Female ; Health Care Costs ; statistics & numerical data ; Hepatitis B ; economics ; Hepatitis B virus ; Hepatitis B, Chronic ; economics ; Hospitalization ; economics ; Humans ; Liver Cirrhosis ; economics ; Liver Neoplasms ; economics ; Male ; Middle Aged ; Surveys and Questionnaires ; Young Adult

The historical evolution， fermentation technology and key links of Sojae Semen Praeparatum （SSP） were sorted out by consulting ancient books and modern literature， and the influencing factors and control methods of quality were analyzed and summarized in order to provide reference for the quality control of SSP. After analysis， it was found that in the fermentation process of SSP， fermentation strains， miscellaneous bacteria， temperature and humidity were all important factors affecting the quality of SSP. The condition control of "post fermentation" process has been paid more attention to in the past dynasties. In addition， the delicious SSP recognized in ancient times should be made from mold fermentation， and the breeding and application of fermented mold may be the key point to solve the quality problem of SSP. Therefore， based on the evaluation indexes of SSP in the past dynasties， it is of great significance to study and optimize the technological conditions such as strain， temperature and humidity in depth to improve the quality of SSP.