Objective To investigate the clinical value of serum VEGF levels in the diagnosis and treatment of colon cancer.Methods Serum VEGF was detected by ELISA,and CEA and CA199 concentration were detected by CLEIA in 66 patients with colon cancer,55 patients with colon benign diseases and 50 health persons.The value of individual and joint detection for VEGF in colon cancer was evaluated.Analysis had been done on relationships between serum VEGF and pathology,treatment effects and prognosis.Results The levels of serum VEGF in colon cancer group [(318.5±148.6) ng/L] were significantly higher than those in control group [(125.7±49.4) ng/L] and benign colon diseases [(136.9±52.6) ng/L] (t =8.830,8.805,all P ＜ 0.01).There was a positive correlation between serum levels of VEGF and depth of tumor size,tumor invasion,lymph node metastasis and TNM stage (P ＜ 0.01).The susceptibilities of VEGF,CEA,CA199 were 61％,45 ％,53 ％.The sensitivity of detection was improved to 86 ％ when the combined detection of VEGF,CEA and CA199 (x2 =11.237,P ＜ 0.01).The serum levels of VEGF in patients with colon cancer was significantly decreased after treatment in the 3,7,10 day compared with that before operation [(272.3±88.1),(236.8±77.4),(173.1±59.9) vs (318.5±148.6) ng/L,t=2.173,P ＜ 0.05; t =3.961,P ＜ 0.01; t=7.464,P ＜ 0.01],respectively.Conclusion The VEGF was related to the onset and progression and metastasis of colon cancer.It has clinical significancy for diagnosis of colon cancer and judgment of curative effect and prognosis.

Objective: To evaluate the inhibition of human endostatin on tumor growth and metastasis of lung adenocarcinoma LA795 in mice. Methods: Recombinant human endostatin was purified from pCX expressed endostatin clones. Plasminogen was purified from outdated human plasma by affinity chromatography, and human angiostatin was produced from human plasminogen digested by elastase and purified by affinity chromatography. LA795 cells were inoculated subcutaneously into the dorsa of T739 mice, and the mice were randomized into 3 groups. From the 1Oth day, the first group was given 20 mg/kg of recombinant human endostatin s.c. qd, the second was treated daily s. c. of 7.5 mg/kg of human angiostatin, and the third group received daily s.c. with equal volumes of PBS for 14 days. Volumes of the subcutaneous tumors, lung weights, the number of lung surface metastases and mice life span were observed. The results were analyzed by q-test. Results: The tumor volumes of both the 1 st and the 2nd groups increased slowly. From the 8th day after being treated, the tumor volumes were decreasing. However, in the 3rd group, the tumor volumes increased continuously. The lung weight and the number of lung surface metastases of the 1st and 2nd groups were less than that of the 3rd group. The average survival periods of the 1st and 2nd groups were longer than that of the 3rd group. Conclusion: Human endostatin and angiostatin have strong inhibitory effects both on growth of primary tumor and metastasis of lung adenocarcinoma LA795, and prolongs the survival period of the tumor-bearing mice.

Objective To study the diagnosis and treatment of neuroendocrine carcinoma of the nasal cavity and paranasal sinuses, and analyse the influencing factors of suvival and prognosis. Methods We retrospectively analyzed the diagnosis and treatment process of 14 patients with sinonasal neuroendocrine carcinoma (SNEC) admitted in The First Affiliated Hospital of Kunming Medical University from 2007 to 2011. All patients were followed up to learn the survival status of them.Results All patients were followed up for one year up to six years except 2 patients who gave up treatment. Five patients died and six survived with good tumor control in the followed up period. Two patients received only endoscopy surgery, and one of them died from lung metastasis in 21 months after operation, and the other one survived with good tumor control, the disease free survival (DFS) was 9 months . Eight patients were treated by endscopy surgery and /or chemo-radiotherapy, three cases died in following-up period, and five of them survivied with good tumor control, and the disease free survival was 20.25 months.Two patients with transcatheter arterial infusion chemotherapy survivied with good tumor control within the follow up period,and the DFS was 25.5 months.Five patients had moderately differentiated SNEC and DFS was 25.5 months. Seven patients had poorly differentiated SNEC with DFS 14.6 months. Six patients were T4N0M0, four patients were T3N0M0, two patients T2N0M0, and their DFS were 19 months, 12.8 months and 33 month, respectively. Conclusions Surgery with radiotherapy and/or chemotherapy is the current treatment method for sinonasal neuroendocrine carcinoma. Small cell neuroendocrine carcinoma with poor differiation displays highly aggressive and poor prognosis. Diagnosis and treatment in early stage is important for good prognosis.

Objective To investigate the homology of methicillin-resistant Stphylococcusaureus(MRSA)from the neonatal intensive care unit(NICU)of a children's hospital,and evaluate routes and preventive strategies of MRSA healthcare-associated infection(HAI). Methods MRSA strains from neonates and environment of NICU between October and December 2014 were collected,and strains were identified by VITEK-2 microbial analysis system and cefoxitin Kirby-Bauer method,homology of MRSA was analyzed by pulsed-field gel electrophoresis (PFGE ). Results A total of 6 MRSA strains were isolated from NICU between October and December 2014,3 of which (bed-58,70,and 100)were detected MRSA from specimens,MRSA were isolated from neonatal incubator and nurse (nasal swabs and hands)who cared for neonate at bed 58. 5 of 6 MRSA strains were homology,antimicrobial susceptibility testing result showed that No. 1-5 strains were resistant to clindamycin and amoxicillin/clavulanic acid,No. 6 strain was slightly different from No. 1-5 strains,No. 6 strain was susceptible to both clindamycin and amoxicillin/clavulanic acid. PFGE results showed that No. 1-5 strains were of the same type,No. 6 strain was a different type. Conclusion The main route of this MRSA transmission is contact transmission,especially through the hands of health care workers,identification and analysis of epidemic strains by PFGE technique is an effective measures to prevent HAI outbreak and perform epidemiological study.

OBJECTIVETo study the feasibility of supercritical fluid extraction (SFE) for arctiin from the fruits of Arctium lappa.

METHODThe extracts were analyzed by HPLC, optimum extraction conditions were studied by orthogonal tests.

RESULTThe optimal extraction conditions were: pressure 40 MPa, temperature 70 degrees C, using methanol as modifier carrier at the rate of 0.55 mL x min(-1), static extraction time 5 min, dynamic extraction 30 min, flow rate of CO2 2 L x min(-1).

CONCLUSIONSFE has the superiority of adjustable polarity, and has the ability of extracting arctiin.

Arctium ; chemistry ; Chromatography, High Pressure Liquid ; Chromatography, Supercritical Fluid ; methods ; Fruit ; chemistry ; Furans ; analysis ; isolation & purification ; Glucosides ; analysis ; isolation & purification ; Plants, Medicinal ; chemistry

Sugarcane stem is an ideal organ for producing foreign pharmaceutical proteins and chemicals by genetic engineering.A perfect promoter driving foreign gene to express strongly and specifically in sugarcane stem is necessary for this purpose.In order to isolate a Sugarcane stem-specific promoter,a fragment of 1968bp nucleotide sequence(Ppst2a)upstream 5' of sugarcane pst2a gene,which was demonstrated to express specifically in sugarcane stem previously was isolated by using chromosomal walking.Bioinformatical analysis of this sequence shows that the sequence contains some typical elements of a promoter.To identify the stem-specific of this promoter,a construct was derived from pCAMBIA1301,which original CaMV 35S promoter was replaced by the 1968bp nucleotide sequence,and named as pCAMBIA1900.Transformations of pCAMBIA1900 and pCAMBIA1301 to leaves and stem pieces of sugarcane were carried out by using particle bombardment.The transient expression of gus showed that the gus expressed specifically in sugarcane with a little higher level compared with CaMV 35S.It is the first report that pst2a promoter is a potential stem-specific promoter which can further be used in transgenes into sugarcane.

Aim The relative bioavalability of hydrochloride eperisone granule in 10 healthy volunteers was studied. Methods The time-plasma concentrations of hydrochloride eperisone granule, as test drug, and myonal, as reference drug, were determined by GC-MS, with tolperisone senuing as internal standard.The pharmacokinetic parameters of both reference and test drug were calculated and analyzed with two-one side test and confidential interval test. Results The results showed that the AUC0-8, AUC0-∞, Cmax, Tpeak, t1/2(?) and t1/2(?) were (17.9?1.3)ng?h?ml-1 and(18.6?1.6)ng?h?ml-1, (19.1?1.2)ng?h?ml-1 and (20.2?1.6)ng?h?ml-1, (5.2?0.5)ng?ml-1 and (5.4?0.5) ng?ml-1, (1.05?0.18)h and (1.08?0.23)h, (0.78? 0.13)h and ( 0.82?0.14)h,( 1.8?0.3)h and (1.8?0.3)h, respectively. The relative bioavalability of test drug was (105? 5)%. Conclusion It can be concluded that the test and reference are bioequivalented between individuals, preparations and periods.

In order to study the feasibility of E2 gene fragment of hepatitis virus G(HGV) as a component of DNA vaccine against the hepatitis virus G infection, a 559bp DNA fragment encoding HGV E2 was cloned into plasmid pCMV-S from pThioHis-E2 in the same reading frame with HBsAg gene to form a recombinant plasmid named pCMV-S-E2. BALB/c mice of Kunming strain were immunized with purified plasmid DNA of pCMV-S-E2 by intra-muscularly inoculation. The immunizations were boosted twice at an interval of 14 days. The whole blood was collected from mice orbit on the day-8 after the last boost. Mice sera were screened by ELISA to determine the humoral immune response using E2-GST fusion protein as the immobilized antigen and the sera from mice immunized with pCMV-S as control. The result indicated that the immunization with plasmid DNA of pCMV-S-E2 could induce quite strong humoral immune response.
Animals ; Female ; GB virus C ; immunology ; Hepatitis Antibodies ; blood ; Mice ; Mice, Inbred BALB C ; Plasmids ; Recombinant Fusion Proteins ; biosynthesis ; immunology ; Vaccines, DNA ; immunology ; Viral Envelope Proteins ; genetics ; immunology ; Viral Hepatitis Vaccines ; immunology

OBJECTIVETo investigate the morphologic and functional characteristics of the immortalized human liver sinusoidal endothelial cell line (LSEC line).

METHODSImmunofluorescence staining and fluorescence microscopy were used to detect the classic endothelial cell markers in LSEC line, and flow cytometry was used to analyze the purity of the human LSEC line. The morphology (including W-P bodies and surface fenestrations) and phagocytotic capacity of the human LSEC line were observed by transmission and scanning electron microscope. The proliferation curve of the human LSEC line was analyzed by MTT assay. The functional differences between the human LSEC line and human primary LSEC in expression of ELAM-1 and ICAM-1, activities of fibrinolysis (PAI-1, t-PA, u-PA), releasing of IL-6 and IL-8 were compared respectively by enzyme linked immunosorbent assay. Comparison of the susceptibility to hypoxia-reoxygenation induced apoptosis between the human LSEC line and human primary LSEC were investigated by TUNEL.

RESULTSThe established human LSEC line maintained a high proliferative ability and has been passaged for more than 80 times in the absence of any growth factors. Immunofluorescence staining showed that the human LSEC line could express classic endothelial cell marks including von Willebrand Factor (vWF), and could take up acetylated low-density lipoproteins (Ac-LDL). The purity of the human LSEC line was confirmed over 95% by flow cytometric analysis. The W-P bodies and the phagocytosis of Dynabeads was demonstrated by transmission electron microscope. And fenestrations could be found cellular surface with scanning electron microscopy. When compared with human primary LSEC, the human LSEC line has an equivalent responsiveness to tumor necrosis factor in up-regulation of ELAM-1 and ICAM-1. The human LSEC line can also release PAI-1, t-PA, u-PA but can not release IL-6 and IL-8 to TNF-alpha. In contrast, human primary LSEC could release IL-6. The human LSEC line showed higher susceptibility to hypoxia-reoxygenation-induced apoptosis, and the percentage of apoptotic cells was as high as (38.4 +/- 6.7)%, while (28.6 +/- 4.5)% and (7.8 +/- 1.2)% respectively in primary LSEC and in human umbilical vein endothelial cells.

CONCLUSIONSThe established human LSEC line maintains the special phenotypes and the major functional characteristics, and especially maintains the high susceptibility to hypoxia-reoxygenation-induced apoptosis. Therefore it is feasible to use this cell line for the study of liver ischemia-reperfusion injury.

Apoptosis ; Cell Line ; Cell Proliferation ; E-Selectin ; metabolism ; Endothelial Cells ; cytology ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Liver ; cytology

Objective To discuss the combined value of gray-scale ultrasound ( GSUS) and contrast-enhanced ultrasound (CEUS) in the diagnosis of benign and malignant thyroid lesions and to explore the optimal diagnostic point of scoring method .Methods Ultrasound images of 178 thyroid lesions confirmed by pathology were synthetically reviewed by scoring 5 GSUS indicators including shape , orientation, interior echogenicity, halo sign, microcalcification and 6 CEUS indicators including relative arrival time of microbubbles in the periphery and interior, peak periphery and interior echogenicity, peripheral ring-enhancement, and homogeneity of enhancement .One positive indicator scored one point .The optimal diagnostic points and their clinical value were explored according to ROC curves .Results Scores of GSUS, CEUS and the combination of GSUS and CEUS were significantly different (Z =10.188,9.843,10.705,all P <0.001). Areas under ROC curves of GSUS, CEUS and the combination of GSUS and CEUS were 0.936, 0.919 and 0.964, respectively.Three or more positive GSUS indicators of five in a thyroid lesion predicted that the thyroid lesion was malignant , with the sensitivity of 79.6% and the specificity of 91.2%.Two or more positive CEUS indicators of six in a thyroid lesion predicted that the thyroid lesion was malignant , with the sensitivity of 91.8% and the specificity of 81.2%.Five or more positive GSUS and CEUS indicators of eleven in a thyroid lesion predicted that the thyroid lesion was malignant , with the sensitivity of 93.6%and the specificity of 92.3%.The areas under the ROC curve of GSUS and CEUS were 0.936 and 0.919.The area under the ROC curve of the combination of GSUS and CEUS was 0.964, larger than the areas under the GSUS ROC curve and the CEUS ROC curve.Conclusion Ultrasound is valuable in the differential diagnosis of benign and malignant thyroid lesions , and the combination of GSUS and CEUS is the most valuable with 5 points as the optimal diagnostic scoring method .