Objective To establish reverse transcriptase-polymerase chain reaction(RT-PCR) with primers specific for CEA gene and CK20 gene to detect circulating tumor cells in peripheral blood of esophageal cancer pa-tients,and try to find the relationship between the mRNA expression and micrometastasis. Methods The expressions of CEA,CK20 were analyzed by RT-PCR in 53 cases of esophageal tumor tissue and in peripheral blood,compared with 10 patients with benign esophageal disease and 20 healthy volunteers. Results The expressions of CEA-mRNA, CK20-mRNA were 96.23% (51/53), 100% ( 53/53 ) in 53 esophageal tumor tissue and were 52.83% (28/53), 49.06% (26/53) in peripheral blood of 53 esophageal cancer patients. In control group,there was only one expression of CEA-mRNA in peripheral blood of 10 patients with benign esophageal disease,as well as in 20 healthy volunteers. There was no expression of CK20-mRNA in peripheral blood of 10 patients with benign esophageal disease and 20 healthy volunteers. Conclusion CEA-mRNA, CK20-mRNA might be specific and sensitive markers to detect circu-lating tumor cells in peripheral blood and their expression was closely related to TNM stages of the esophageal cancer patients.

Objective To investigate the expressions and clinical significances of MUC1-mRNA and CK20-mRNA in peripheral blood of esophageal cancer patients.Methods MUC1-mRNA and CK20-mRNA were detected in 53 patients with esophageal cancer,10 patients with esophageal benign tumor and 20 healthy volunteers by RT-PCR technique.Results The expressions of MUC1-mRNA,CK20-mRNA and combining group were 35.85 % (19/53),49.06 % (26/53) and 62.26 % (36/53) in peripheral blood of 53 esophageal cancer patients.In control group,there was no expression of MUC1-mRNA and CK20-mRNA in peripheral blood of 10 patients with benign esophageal disease and 20 healthy volunteers.The positive rate increased by combining test(x2 =11.0228,P ＜0.05).Conclusion MUC1-mRNA and CK20-mRNA might be specific and sensitive markers to detect circulating tumor cells in peripheral blood and their expressions are closely related to TNM stages of the esophageal cancer patients.The combining test might be of high value of the diagnosis of micrometastasis.

Background: There is no gold standard for the diagnosis and monitoring of inflammatory bowel disease (IBD).As immune system plays crucial role in the pathogenesis of IBD,some immune-specific serum antibodies are considered to be useful tools for the diagnosis and differential diagnosis of the disease.Aims: To investigate the clinical significance of serum antibodies,including anti-Saccharomyces cerevisiae antibody (ASCA) and perinuclear anti-neutrophil cytoplasmic antibody (pANCA) in IBD.Methods: Serum samples were obtained from 91 consecutive IBD patients from Feb.2015 to May 2016 at the First Affiliated Hospital of Soochow University.Of them,52 were Crohn's disease (CD) and 39 were ulcerative colitis (UC).Serum samples of 36 non-IBD patients were served as controls.ASCA-IgG and ASCA-IgA were detected by ELISA,and pANCA by indirect immunofluorescence assay.Using clinical diagnosis as gold standard,crosstabs statistics was performed to measure the diagnostic accuracy of ASCA and pANCA,and ROC curve,Pearson Chi-square test and Fisher's exact test were employed for analyzing the correlations of these two serum antibodies with IBD,CD,UC,and the location of the disease.Results: Both serum ASCA-IgG and IgA were correlated with CD (AUC=0.626 and 0.614),while UC was correlated with ASCA-IgA only (AUC=0.486).Serum pANCA had relevance to IBD (r=0.342),CD (r=-0.262) and UC (r=0.614);its sensitivity and specificity for IBD and UC were superior to those for CD (P＜0.05).In CD patients,ASCA-IgG was associated with terminal ileal disease (P＜0.05),and pANCA was associated with colonic involvement (P＜0.05).In UC patients,both ASCA-IgG and IgA were correlated with terminal ileal disease (P＜0.05).Conclusions: Serum ASCA and pANCA may be helpful for discrimination between CD and UC when the diagnosis of IBD is established.Furthermore,they are closely associated with the disease location,ASCA is related with the terminal ileal disease and pANCA is related with colonic involvement.

Objective:To investigate the clinical efficacy of intramedullary nailing combined with bone cement in repair of segmental bone defects after tumor resection.Methods:A retrospective analysis was conducted of the 5 patients with malignant bone tumor who had been treated at Department of Orthopaedics, Qiannan People's Hospital from April 2018 to September 2019 for remaining segmental bone defects following limb salvage surgery. They were 4 males and one female, aged from 11 to 55 years (average, 35.4 years). Their defects ranged from 6 to 21 cm (average, 12.3 cm) after tumor resection. By the Karnofsky performance score (KPS) for long-term quality of life, all of them scored less than 50 points. Of them, 3 were treated by interlocking intramedullary nails and bone cement filling, and 2 by elastic intramedullary nails and bone cement filling. In the 2 cases with defects of 21 cm and 13 cm, the fixation was assisted by a plate and an external fixator. Defect length after resection, operation time and intraoperative bleeding were recorded; the efficacy was evaluated by the Enneking functional evaluation of reconstructive procedures after surgical treatment of tumors of the musculoskeletal system, visual analogue scale (VAS), and KPS.Results:All the 5 patients had uneventful surgery, with operation time ranging from 112 to 225 min (average, 154.2 min), intraoperative bleeding from 300 to 500 mL (average, 382 mL), and defect length after resection from 6 to 21 cm. The 5 patients were followed up for 6 to 28 months. Of them, 2 died of disease progression 6 and 7 months after surgery, respectively. According to the Enneking's evaluation, one patient scored 28 points, 2 patients 23 points and 2 patients 21 points, giving a high degree of satisfaction. Their VAS scores 6 months after surgery ranged from 1 to 6, averaging 3.6; their postoperative KPS scores ranged from 60 to 80, averaging 72.Conclusion:In repair of segmental bone defects after tumor resection, intramedullary nailing combined with bone cement filling can relieve pain of patients and lead to satisfactory short-term curative efficacy.

OBJECTIVETo determine whether B7-2 could improve hepatitis B virus (HBV)-specific immunity induced by inoculation of a plasmid encoding HBV preS2+S HBV DNA vaccine.

METHODSB7-2 expression plasmids were inoculated with the DNA vaccine in the gastrocemius muscles. Then the cytolytic T Lymphocyte activity (CTL), the delayed-type hypersensitivity (DTH) response and the titers of anti-HBs were analysed.

RESULTSThe DTH response and CTL activity were significantly enhanced when B7-2 expression plasmids were coinoculated with the DNA vaccine (P<0.01), but the titers of anti-HBs were unaffected (P>0.05).

CONCLUSIONSThe results suggest that B7-2 expression plasmid could improve HBV specific cell-mediated immunity induced by HBV DNA vaccine, and the gene-based coinoculation strategy using HBV viral antigen and B7-2 costimmulating could be a powerful means of combating HBV infection.

Adjuvants, Immunologic ; Animals ; Antigens, CD ; immunology ; B7-2 Antigen ; Hepatitis B Antibodies ; biosynthesis ; Hepatitis B Vaccines ; immunology ; Hypersensitivity, Delayed ; immunology ; Male ; Membrane Glycoproteins ; immunology ; Mice ; Mice, Inbred C57BL ; Plasmids ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccines, DNA ; immunology

AIM: To investigate the inhibitory effect s of a synthetic CRE-transcription factor decoy oligodeoxynucleotide (CRE-decoy ODN) on the upregulation of the expression of cholecystokinin (CCK) and fosB mRN A induced by chronic morphine administration in SK-N-SH cells. METHODS: The CRE cis-element, TGACGTCA, was palindromic, a sy nthetic single-stranded phosphorothioate oligodeoxynucleotide composed of the CR E sequence self-hybridizes to form a duplex/hairpin. The CRE-palindromic decoy a nd control oligodeoxynucleotides were added to the medium (1 h before exposure t o morphine) at 150 nmol/L in the presence of cationic lipid DOTAP. After the cel ls were treated with 100 ?mol/L morphine for 48 h, 10 ?mol/L naloxone was use d for 15 min. The effects of CRE-decoy ODN on the DNA-binding activity of CREB, the expression of CCK and fosB mRNA were detected by electrophoresis mobi lity shift assay (EMSA) and RT-PCR, respectively. The stability of cell-incorpo rated [ 32P]-labeled CRE-decoy ODN was extracted with phenol:chloroform a nd then subjected to 20% nondenaturing polyacrylamide gel electrophoresis and au toradiography. RESULTS: Chronic morphine administration and acute naloxone-prec ipitated withdrawal significantly activated the DNA-binding activity of CREB and the expression of CCK and fosB mRNA in SK-N-SH cells. The CRE-decoy ODN pen etrated into the cells, specifically downregulated these indexes. CONCLUSIONS: CRE-decoy ODN can significantly downregulates the e xpre ssion of CCK and fosB mRNA through specifically suppressing the DNA-binding activity of CREB activated by chronic morphine administration in SK-N-SH cells.

AIM: To investigate the effect of epinephrine on LPS-induced pro-inflammatory mediators (TNF-?, NO and COX-2) and anti-inflammatory mediators (HO-1 and IL-10) production in murine macrophage RAW264.7 cells, and to determine whether these effect is due to the influence of epinephrine on NF-?B activation. METHODS: RAW264.7 cells were cultured in vitro with 10 ?g/L LPS in the absence or presence of epinephrine at variant concentrations (1, 5, 10, 50 ?mol/L) for 24 hours, then the supernatants was collected for measuring TNF-? and IL-10 by ELISA and Griess reagent was used to measure NO (NO_2-/NO_3-) concentration. At the same time point, cells were harvested and COX-2, HO-1 and I?B-? was detected by Western blotting. RESULTS: 10 ?g/L LPS significantly induced the production of TNF-?, NO (NO_2-/NO_3-), COX-2, HO-1 and IL-10. When epinephrine was added into the medium together with LPS, the pro-inflammatory mediators production was decreased in a dose-dependent manner, however, anti-inflammatory mediators HO-1 and IL-10 expression was enhanced by epinephrine. Epinephrine has no significant effect on I?B-? degradation in LPS-activated RAW264.7 cells. CONCLUSION: Epinephrine down-regulates LPS-induced pro-inflammatory mediator expression while promotes anti-inflammatory mediator production in murine macrophages. These effect seems to be independent of NF-?B activation.

OBJECTIVETo prepare and evaluate the injectable temperature responsive in situ gel for sustained release of Glabrous Sarcandra Herb extract in vitro.

METHODThe novel temperature responsive poloxamer 407 was used as the carrier material of Glabrous Sarcandra Herb extract sustained release injection. The release of Glabrous Sarcandra Herb extract from poloxamer 407 based in situ gel in the phosphate buffer (pH 7. 6) was studied at 37 degrees C under agitation. HPLC method was used for the determination of Glabrous Sarcandra Herb extract.

RESULTThe best prescription was composed of by 16% P-407, 6% P-188, 0.2% HPMC, 0.5% benzil alcohol and 4% (g mL(-1)) Glabrous Sarcandra Herb extract. The optimal gelation temperature was around 33 degrees C. The data of release in vitro were analyzed according to the theoretical model of Korsmeyer-Peppas.

CONCLUSIONThe preparation method of the injectable thermosensitive in situ gel of Glabrous Sarcandra Herb extract is appropriate. The release in vitro can reach the expecting request.

Ferns ; chemistry ; Plant Extracts ; analysis ; Poloxamer ; chemistry ; Temperature

OBJECTIVETo study the preparation ofnanostructured lipid carriers loaded with supercritical carbon dioxide fluid extraction of Xionggui powder.

METHODPrepared nanostructured lipid carriers (NLC) loaded with supercritical carbon dioxide fluid extraction of Xionggui powder (XG-CO2-SFE) with a microemulsion ultrasonication method , established the best prescription of XG-CO2-SFE-NLC by orthogonal design methods with entrapment efficiency of nanoparticles as index, and investigated their physicochemical characterizations.

RESULTThe best prescription was m(phospholipid) : m(S-40) : m(lipid carriers) = 5 : 2 : 1, the entrapment efficiency of nanopartices was 86.8%, results revealed that nanoparticles were sphere like with the mean size of 92.6 nm and the mean zeta potential was -20.68 mV.

CONCLUSIONThe prep aration method of the XG-CO2-SFE-NLC is appropriate.

Carbon Dioxide ; chemistry ; Chromatography, Supercritical Fluid ; Drug Carriers ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Emulsions ; Linear Models ; Lipids ; chemistry ; Nanostructures ; chemistry ; Powders