This study aimed to identify the related substances of phloroglucinol injection by two-dimensional liquid chromatography quadrupole time-of-flight mass spectrometry (2D-LC-Q-TOF/MS). The first-dimensional separation was carried out on an HSS T3 (250 mm × 4.6 mm, 5 μm) column by gradient elution using 1.36 g·L^-1 potassium dihydrogen phosphate buffer solution (pH adjusted to 3.0 with diluted phosphoric acid) and acetonitrile as the mobile phases. The separated components were then trapped in switch valve tube lines respectively and delivered to the second-dimensional desalting gradient elution which was performed with a BDS C18 (100 mm × 4.6 mm, 2.4 μm) column using 0.1% formic acid and methanol as the mobile phases. After rapid desalting, electrospray-ionization quadrupole time-of-flight high resolution mass spectrometry was used for determining the accurate masses and elemental compositions of the parents and their product ions for both phloroglucinol and its related substance. Structures of the related substances were then figured out by mass spectrometry elucidation, organic reaction mechanism analysis, and/or comparison with reference substances. Under the established analytical conditions, phloroglucinol and its related substances were adequately separated, 17 main related substances were detected and identified in the injection and its stressed samples for the first time. The identification results can provide reference for the quality control of phloroglucinol injection.

Objective To investigate the effect of silencing E6-associated protein(E6AP)on the level of p53 protein in human papilloma virus(HPV)negative cervical cancer cells(C33A cells).Methods The siRNA sequence silencing E6AP(siE6AP)and silencing control disordered siRNA sequence(siControl)were transfected into C33A cells with the mediation of LipofectamineTM2000 transfection reagent respectively.The silencing effect of siRNA on E6AP and the expression of p53and cleaved-caspase-3 proteins were detected by Western blot.Results The levels of E6AP protein in C33A cells of siE6AP group were significantly lower(t =-4.597,P<0.05),while the levels of p53 and cleaved-caspase-3 proteins were significantly higher than those of siControl group(t = 4.533 and 7.099 respectively,each P<0.05).Conclusion Silencing of E6AP significantly increased the expression of p53 protein in C33A cells,indicating that silencing of E6AP may restore the activity and function of p53 protein in C33A cells.

This study aims to investigate the function of two SNPs (rs8904C > T and rs696G >A) in 3' untranslated region (3'UTR) of NFKBIA gene by constructing luciferase reporter gene. A patient's genomic DNA with rs8904 CC and rs696 GA genotype was used as the PCR template. Full-length 3'UTR of NFKBIA gene was amplified by different primers. After sequencing validation, these fragments were inserted to the luciferase reporter vector, pGL3-promoter to construct recombinant plasmids containing four kinds of haplotypes, pGL3-rs8904C/rs696G, pGL3-rs8904C/rs696A, pGL3-rs8904T/rs696G and pGL3-rs8904T/rs696A. Then these plasmids were transfected into LS174T cells and the luciferase activity was detected. Compared with pGL3-vector transfected cells (negative control), the luciferase activity of the four kinds of recombinant plasmids was significantly decreased (P < 0.001). For rs696G > A, the luciferase activity of the recombinant plasmids containing A allele (pGL3-rs8904C/rs696A and pGL3-rs8904T/rs696A) was about 45.1% (P < 0.05) and 56.1% (P < 0.001) lower than those containing G allele (pGL3-rs8904C/rs696G and pGL3-rs8904T/rs696G), respectively. For rs8904C > T, there were no significant differences in the luciferase activity between the recombinant plasmids containing T allele and those with C allele. Together, the luciferase reporter gene vectors containing SNPs in NFKBIA gene 3'UTR were constructed successfully and rs696G > A could decrease the luciferase activity while rs8904C >T didn't have much effect on the luciferase activity.
3' Untranslated Regions ; Genes, Reporter ; Genetic Vectors ; Humans ; I-kappa B Proteins ; genetics ; Luciferases ; NF-KappaB Inhibitor alpha ; Plasmids ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; Transfection

BACKGROUNDTo study apoptosis of peripheral blood cells of children with viral pneumonia, explore immunopathogenesis and the possibility of immunotherapy of patients with viral pneumonia.

METHODSFresh peripheral blood samples were collected from 28 patients with viral pneumonia and 24 healthy children were treated and run through the flow cytometry. The data were acquired using Cell Quest software and the percentage of live cells, viable apoptotic cells, non-viable apoptotic cells and dead cells of neutrophils and lymphocytes were counted. The patients with viral pneumonia were hospitalized at our hospital. The average age of patients was 1.3 years; 24 healthy children were served as control group (age 1.8 years, on average). T-test and variance analysis by SPSS FOR WINDOWS 10.0 software was used for statistical analysis.

RESULTSThe percentage of live neutrophils and lymphocytes in the acute stage and recovery stage in patients were significantly lower than that in control group (P < 0.01). The percentage of viable apoptotic neutrophils and lymphocytes in two stages in patients were significantly higher than that in control group (P < 0.05). Except for the percentage of live cells, non-viable apoptotic cells and dead lymphocytes, others had no difference between the patients and control groups.

CONCLUSIONApoptosis of neutrophils and lymphocytes of peripheral blood cells of children with viral pneumonia increased. Whereas the percentage of live cells decreased. Drugs that can accelerate apoptosis may be helpful in treatment of viral pneumonia.

Apoptosis ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Infant ; Lymphocytes ; pathology ; Male ; Neutrophils ; pathology ; Pneumonia, Viral ; pathology

0.05).Moreover,the growth curves of the two kinds of cells were similar.Con- clusions The cell growth properties of cultured transplanted rabbit SMG are similar to that of normal SMG,the cytobiological charac- teristic of transplanted autologous free rabbit SMG are not changed evidently.

Objective To investigate the effects of hydrogen sulfide(H2S)on vascular inflammation and blood pressure in spontaneously hypertensive rats(SHR).Methods Four weeks old male SHR rats were treated with saline(control,n=7),sodium hydrosulfide(NaHS,a H2S donor,n=7)and propargylglycine(PPG,endogenous H2S production inhibitor,n=6) for 5 weeks.Age-natched male Wistar Kyoto(WKY)rats served as normotensive controls(n=8).Five weeks later,systolic blood pressure (SBP) was measured in conscious and quiet rats by means of the standard tail-cuff method.The protein expressions of intereellular adhesive molecule-1(ICAM-1),nuclear transcriptional factor-κB p65(NF-κB p65)and inhibitor of nuclear transcriptional factor-κB(IκB-α) in thoracic aorta of rats were detected by immunohistochemieal assay.while the expression of ICAM-1 mRNA in thoracic aorta of rats were investigated by in situ hybridization.Results SBP of control SHR rats was significantly hisher than that of WIKY rats(P<0.05)accompanied by significantly upregulated expressions of ICAM-1 mRNA,ICAM-1 protein,NF-κB p65 protein in aortic endothelial cells(all P<0.01),while the expression of IkB-a protein in aortic endothelial cells in SHR control group was significantly lower than that of WKY control group(P<0.01).NaHS treated SHR rats showed significantly reduced SBP and downregulated expressions of ICAM-1 mRNA,ICAM-1,NF-κB p65 in aortic endothelial cells and upregulatod expression of IκB-α protein in aortic endothelial cells compared to untreated control SHR rats(all P<0.05).In SHR rats treated with PPG.the expressions of ICAM-l mRNA,ICAM-1 protein,NF-κB p65 protein in aortic endothelial cells were further increased while the expression of IκB-α protein further decreased compared with control SHR rats(all P<0.05).Conclusion H2S might attenuate the development of hypertension through by attenuating vascular inflammation reactions.

Objective To explore the effect of individual health education on the patients with colostomy.Methods Seventy-eight patients received individual health education.They were investigated with the questionnaire about the exercise of selfvcare-agency(ESCA) and the assessment of self-care agency in third to fifth day after operation,left hospital and three months after discharged.Results The average score of ESCA of patients in third to fifth day after operation,left hospital,three months after discharged was ( 96.63 ± 10.13 ),(106.18 ± 8.94),(134.74 ± 9.54),and different time slot with different score,and the different was statistically different (P ＜ 0.01 ),and the sub-scale score of self-care skill,self-nursing sense,self conception,health knowledge level,colostomy care was significantly different between different time slots ( P ＜ 0.01 ).Conclusions Individual health education can obviously improve the self-care agency of patients with colostomy and effectively reduce the complication of colostomy.

Objective: To study the inhibitory effect of Kangxian Recipe (KXR) on TGF-β1 induced hepatocyte apoptosis. Methods: The in vitro model of hepatocyte apoptosis was established by cell biologic methods, utilizing the characteristics of TGF-β1 to observe the inhibitory effect of KXR on hepatocyte apoptosis. Results: TGF-β1 induced apoptosis of hepatocyte in a dose-dependent manner. The apoptosis rate of 2.2.15 cells was 63% when 500 ng/L TGF-β1 was used, while for HepG2 cell, it was merely 44%. After treatment of 20 ng/L KXR, the apoptosis rate of the two kinds of cell lines lowered to 33% and 24% respectively. The inhibiton rate of both groups was about 50%. Conclusion: KXR had strong inhibitory effect on hepatocyte apoptosis induced by TGF-β1.

OBJECTIVETo analyze the feasibility and the value of resection for lung cancer invading the superior vena cava (SVC).

METHODSBetween 1988 and 2005 the data of 31 patients who underwent resection were analyzed retrospectively. The reconstruction was done using simple suture, pericardial patch or prosthetic replacement. Postoperative morbidity, long-term survival were examined using the Kaplan-Meier method (Log rank test) and the COX model for survival.

RESULTSSeventeen squamous cell carcinomas, 8 adenocarcinomas, and 6 undifferentiated small cell carcinomas were resected. There were 13 partial SVC resection, the reconstruction was done using a simple running in 5 patients, and a pericardial patch in 8 patients. Eighteen patients underwent complete resection of SVC with prosthetic replacement. The time of clamping the SVC system was from 8 to 35 minutes for complete resection patients, while the time was from 3 to 15 minutes for partial resection patients. One patient didn't clamp the SVC. Postoperative morbidity and mortality were 48% and 0%, respectively. One, 3 and 5-year survival rates were 61%, 33% and 21%, respectively, with median survival at 31 months. Survival rate of patients with N2 disease was obviously lower than those with localized (N0/N1) nodal disease (chi2 = 14.3, P = 0.000), the median survival was 42 and 13 months respectively. There were no significant effects on overall survival with pathologic features and surgery methods. Survival rate of patients with induction chemotherapy before operation or intraoperative chemotherapy was higher than those received direct surgery (chi2 = 5.0, P = 0.025), the median survival was 39 and 14 months respectively.

CONCLUSIONSThe resection of the SVC for involvement by lung cancer can be performed in selected patients, especially for those with localized (N0/N1) nodal disease. Induction chemotherapy should be performed.

Adult ; Aged ; Blood Vessel Prosthesis Implantation ; Feasibility Studies ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; mortality ; pathology ; surgery ; Male ; Middle Aged ; Pneumonectomy ; methods ; Prognosis ; Retrospective Studies ; Survival Analysis ; Survival Rate ; Vascular Neoplasms ; mortality ; secondary ; surgery ; Vena Cava, Superior ; pathology

This study was purposed to investigate the angiogenin (ANG) expression in COS-7 cells and its biological activity. The gene of angiogenin was obtained from mononuclear cells of peripheral blood by using RT-PCR and inserted into eukaryotic expression vector of pcDNA3.1. After being transfected into COS-7 cells, the recombinant ANG was identified by Western blot assay. The function of promoting proliferation of ANG to ECV304 cells was detected by MTT method, and its activity of vascularization was analyzed by chick embryo chorioallantois treated by the culture supernatant after transfection with pcDNA3.1-ang. The result showed that recombinant ANG was expressed in COS-7 cells after transfection for 24 to 36 hours. It could specifically react with monoclonal antibody against ANG. The recombinant ANG could obviously promote the proliferation of ECV304 cells. In contrast with the control group, the culture supernatant of pcDNA3.1-ang transfected group could stimulate the angiogenesis in embryo chorioallantois. It is concluded that the ang transiently expresses in COS-7 cells, and its expression product obviously stimulates the cell proliferation and angiogenesis.
Angiogenesis Inducing Agents ; pharmacology ; Animals ; COS Cells ; metabolism ; Cell Line ; Cell Proliferation ; drug effects ; Cercopithecus aethiops ; Endothelial Cells ; cytology ; Genetic Vectors ; genetics ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Ribonuclease, Pancreatic ; biosynthesis ; genetics ; pharmacology ; Transfection