On the basis of the Uncaria transcriptome, specific primers were designed for UrSTR. The full-length cDNA of UrSTR (GeneBank: OL310251) was 1 541 bp, encoding 345 amino acid residues, and the promoter region sequence of UrSTR (GeneBank: OL310252) was 1 179 bp. Phylogenetic tree is revealed that UrSTR had a closest relationship with STR from Ophiorrhiza pumila and Ophiorrhiza japonica. Localization of UrSTR protein is revealed located in the vacuole membrane. Plant-care analysis indicated that the promoter region sequence of UrSTR, covering multiple light, stress and hormone-response cis-regulatory elements, and verified transcriptional activity. The results of SDS-PAGE show that pET-28a-UrSTR recombinant protein was successfully expressed, and the size was anticipated. The UrSTR prokaryotic expression system needs to be optimized in the later stage. The research lays the foundation for further purification to study its structure and functional characterization of the UrSTR protein.

OBJECTIVETo establish a modified plasma protamine paracoagulation test.

METHODSPlasma protamine paracoagulation, modified plasma protamine paracoagulation and D-dimer (D-D) tests were performed for the plasma samples collected from 98 cases of disseminated intravascular coagulation (DIC) and 156 normal subjects. The sensitivity and specificity of the 3 tests were analyzed. The plasma samples from 8 cases of suspected myocardial infarction were detected using modified plasma protamine paracoagulation for diagnostic purpose.

RESULTSThe sensitivity of plasma protamine paracoagulation, modified plasma protamine paracoagulation and D-D tests was 16.33%, 88.76% and 77.56%, and the specificity was 100%, 88.46% and 97.44%, respectively. Positive results occurred earlier in modified plasma protamine paracoagulation test than in plasma protamine paracoagulation and D-D tests in 5 cases of myocardial infarction.

CONCLUSIONThe modified plasma protamine paracoagulation test has a higher sensitivity than plasma protamine paracoagulation test and a higher specificity than D-D test, and can be helpful in early diagnosis of thrombosis and fibrinolysis.

Adult ; Blood Coagulation Tests ; methods ; Female ; Humans ; Male ; Middle Aged ; Protamines ; blood ; Sensitivity and Specificity ; Thrombosis ; blood ; diagnosis

OBJECTIVETo explore the effects of mitochondrial pathways on apoptosis in colon carcinoma cells induced by Tumor necrosis factor related apoptosis inducing ligand and offer evidences for TRAIL application in clinic.

METHODSApoptosis, integration of mitochondria (including DeltaPsim, cardiolipin), activity of Caspase-9 and release of cytochrome c in colon carcinoma cells SW1116 treated with TRAIL, were detected by means of flowcytometry, fluorometer method and western-blot at the different time point.

RESULTSAfter treated with TRAIL for 4 hours, the apoptosis index was 32.98%, and the damage of mitochondria occurred with DeltaPsim, cardiolipin decreased, and the activity of Caspase-9 and cytochrome c increased. The Caspase-9 activity at 24 hour was (48.12+/-2.21)micromol.L(-1).h(-1).mg(-1) protein. Mitochondrial damage induced by TRAIL could be inhibited by Caspase inhibitor Z-VAD. fmk.

CONCLUSIONMitochondrial pathways involved in the apoptosis of colon carcinoma cell induced by TRAIL. Cytochrome c was released and Caspase-9 was activated in the Caspase-dependent manner after the damage of mitochondrial.

Apoptosis ; Cardiolipins ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cytochromes c ; metabolism ; Humans ; Mitochondria ; metabolism ; Mitochondrial Membranes ; metabolism ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; metabolism ; TNF-Related Apoptosis-Inducing Ligand ; metabolism

OBJECTIVETo determine the effects of PCMV-FGEV transfection on the profile of SMMC-7721 hepatocellular in vitro in vivo.

METHODSSMMC-7721 hepatocellular was transfected with PCMV-FGEV antisense, PCMV-VEGF sense and empty vector plasmid encapsulated by lipofectamine. The positive cell clones were selected with G418. The stable transfection and expression of VEGF in the SMMC-7721 hepatocellular were determined by in situ hybridization and immunochemical analysis. The effect of PCMV-FGEV transfection on SMMC-7721 hepatocellular proliferation was observed by MTT colorimetric assay. Flow cytometry was used to determine the effects of PCMV-FGEV transfection on cell apoptosis of SMMC-7721. The growth of transfected cells was also observed in nude mice.

RESULTSThere was reduced VEGF expression in SMMC-7721 transfected with PCMV-FGEV confirmed by in situ hybridization and immunohistochemical analysis. There was no effect of PCMV-FGEV transfection on cell proliferation and cell apoptosis of SMMC-7721 in vitro. The growth of cell with PCMV-FGEV transfected was slow in nude mice (vivo) and accompanied with obvious apoptosis. The latent time of tumor in the antisense mice group was 25.0+/-1.8 days, which was longer than that in sense and control group significantly (F=19.455, P<0.01). On the other hand, the average tumor weight in antisense group (0.96 g+/-0.28 g) was the smallest among the three groups (F=21.501, P<0.01).

CONCLUSIONSThe expression of VEGF was inhibited by PCMV-FGEV. There was no effect on cell proliferation and cell apoptosis of SMMC-7721 by transferring PCMV-FGEV gene into SMMC-7721 cells in vitro. But in vivo it can inhibit tumor growth and induce cell apoptosis.

Apoptosis ; Cell Division ; Cell Line, Tumor ; Humans ; Immunohistochemistry ; Liver Neoplasms ; pathology ; therapy ; RNA, Antisense ; therapeutic use ; Transfection ; Vascular Endothelial Growth Factor A ; analysis ; antagonists & inhibitors ; genetics

Bacillus are well known antibiotic producers. In this study,dozens of Bacillus strains from different sources were screened. Among them,a strain with strong antifungal activity was found. With 16S rDNA test and Biolog assay,this strain was identified to be Bacillus amyloliquefaciens. The fermentation conditions were optimized in small conical flasks. After ammonium sulfate salting out,dialysis,freezing vacuum dehydration,the crude protein extracts were obtained. The thermal stability,pH stability,protease stability,ion stability and antifungal spectrum of this protein were studied further. Scanning electronic microscope was also used to explore the antifungal mechanism.

OBJECTIVETo compare the safety and efficacy between two different surgical approaches for thoracoscopic esophagectomy including left lateral decubitus position and prone position.

METHODSFrom January 2008 to December 2009, 88 patients who underwent thoracoscopic esophagectomy were enrolled in this study. Among them, 52 patients were placed in decubitus position and 36 patients were placed in prone position.

RESULTSNo conversion to thoracotomy occurred in either group. The operative time was shorter in the prone group than that in the decubitus group (70 ± 20 min vs. 82 ± 17 min, P<0.01). Blood loss during operation was less in the prone group(100 ± 52 ml vs. 139 ± 54 ml, P<0.01). More lymph nodes were harvested from chest in the prone group(12.2 ± 6.2 vs. 8.6 ± 4.3, P<0.01). There was no significant difference between the two groups in morbidity.

CONCLUSIONThoracoscopic esophagectomy in prone position is associated with better exposure of surgical filed, shorter operative time, less blood loss, and more extensive lymph node dissection as compared to decubitus position.

Aged ; Esophageal Neoplasms ; surgery ; Esophagectomy ; methods ; Female ; Humans ; Male ; Middle Aged ; Posture ; Prone Position ; Retrospective Studies ; Thoracoscopy ; Treatment Outcome

Background and Objective:Cytokine-induced killer(CIK)cells have high anti-tumor activity for hepatocellular carcinoma(HCC).Whether CIK cell therapy can eradicate residual cancer cells and prevent or postpone tumor relapse after transcatheter arterial chemoembolization(TACE)should be testified.This study was to evaluate the efficacy of CIK cell therapy combined with TACE on HCC.Methods:A total of 146 consecutive patients with unresectable HCC were divided into combination group (72 patients trealted with CIK cell therapy combined with TACE) and TACE group(74 patients treated only with TACE).The progression-free survival(PFS)and overall survival(OS)were analyzed.Results:The 6-month,1-year,and 2-year PFS rales were 72.2%, 40.4%, 25.3%in combination group, and 34.8%,7.7%,2.6%in TACE group.The median time to progression was 11 months[95%confidence interval(CI),8-1 4 months]in combination group and 5 months(95% CI,4-7 months)in TACE group.The estimated 6-month, 1-year, and 2-year OS rates were 90.3%, 71.9%. 62.4%in combination group,and 74.6%,42.8%,18.8%in TACE group.The median OS was 31 months(95%CI,27-35 months)in combination group and 10 months(95% CI,7-13 months)in TACE group.The times of TACE,ECOG performance status, and CIK cell therapy were independent prognostic factors for PFS and OS.Conclusion:Adjuvant immunotherapy with CIK cells could greatly improve the efficacy of TACE on HCC,and plays an important role in prolonging the PFS and OS of HCC patients after TACE.

Objective To evaluate the feasibility, safety and efficacy of intra-arterial thrombolytic therapy on elderly patients (≥ 80 years old) with acute ischemic stroke. Methods The clinical data of 86 patients with acute ischemic stroke, received intra-arterial thrombolytic therapy, were retrospectively analyzed; according to age differences, these patients were divided into advanced age group (≥80 years old, n=21) and common age group (<80 years old, n=65); and control group (≥80 years old, not receiving thrombolytic therapy, n=50) was established. The recanalization rate and early clinical improvement rate, and the incidence, recover rate and death rate of symptomatic intracerebral hemorrhage were evaluated in these patients after treatment. Results No significant differences in the favorite recanalization rate and short-term outcome, and the incidence of symptom intracranial hemorrhage were noted between the advanced age group and common age group (P=0.528, P=0.102,P=0.353). The incidence of symptom intracranial hemorrhage in the advanced age group was obviously higher than that in the control group (P=0.034); the recover rate of symptom ntracranial hemorrhage in the advanced age group (42.9%) was obviously lower than that in the common age group (50.8%), but significantly higher than that in the control group (16%, P=0.042, P=0.017). The mortality of the advanced age group was similar to that of the control group (23.8% versus 28%, P=0.816), but higher than that of common age group (23.8% versus 10.8%, P=0.034). Conclusion Relatively high feasibility, safety and efficacy of intra-arterial thrombolytic therapy are noted in elderly patients (≥80 years old) with acute ischemic stroke, demonstrating that the use of intra-arterial thrombolytic therapy in very elderly patients should not be avoided but pursued advisably.

To provide evidence for Campylobacter jejuni (C.jejuni) vaccine research,the B cell epitope of AhpC protein were predicted,and the antigenicity was analyzed.AhpC was found locating in outer membrane of C.jejuni without transmem brane structure and no signal peptide by bioinformatics software TMHMM Server V2.0,SignalP 4.1.There were seven B-cell linear epitopes in AhpC predicted by IEDB.Then,the AhpC protein and chemically synthesized antigenic epitopes of C.jejuni were used as antigens,and the AhpC antibody of C.jejuni was used as the primary antibody,the antigens of predominant linear B cell epitopes were detected by ELISA and dot blot.Results showed that one epitope of B cell epitopes (AhpC4-16) was able to recognize the antibodies of C.jejuni AhpC and had strong antigenicity,indicating that could be used in the follow-up vaccine research.

AIMTo develop an HPLC-MS assay for determination of donepezil in human plasma and to investigate the pharmacokinetics and bioequivalence of donepezil capsule in healthy volunteers.

METHODSA randomized crossover design was performed in 20 healthy volunteers. In the two study periods, a single 5 mg dose of either capsule or tablet was administered to each volunteer. After spiked with the internal standard (phenoprolamine) and treated with saturated sodium bicarbonate, plasma was extracted with ethyl acetate and separated with a C18 reversed phase column. LC-ESIMS was used in the selected ion monitoring (SIM) mode with target ions at m/z 380 for donepezil and m/z 344 for phenoprolamine. The fragmentor voltage was 120 V. The main pharmacokinetic parameters of donepezil and the bioequivalence of its two preparations were calculated.

RESULTSThe main pharmacokinetic parameters T1/2, Tmax and Cmax were (63 +/- 10) h, (3.3 +/- 0.4) h and (8.5 +/- 0.4) microgram.L-1 for the capsule; (57 +/- 9) h, (3.4 +/- 1.0) h and (8.1 +/- 1.0) microgram.L-1 for the tablet, respectively. The relative bioavailability of the donepezil capsule was 102% +/- 11%.

CONCLUSIONThe assay was shown to be sensitive, accurate and convenient. The two preparations of donepezil were bioequivalent.

Adult ; Area Under Curve ; Biological Availability ; Capsules ; chemistry ; Cholinesterase Inhibitors ; administration & dosage ; pharmacokinetics ; Chromatography, High Pressure Liquid ; Cross-Over Studies ; Humans ; Indans ; administration & dosage ; pharmacokinetics ; Male ; Piperidines ; administration & dosage ; pharmacokinetics ; Spectrometry, Mass, Electrospray Ionization ; Tablets ; chemistry ; Therapeutic Equivalency