Objective To open out new method,we probe into immune function of??ST and NKT cells in NSCLC.Methods The peripheral blood cells were stained with antibodies labeled with fluorescence in NSCLC,??T,NKT and their set group cells were counted with flow cytometry.Results The absolute counts of??T,CD~+_(56)??T,CD~-_(56)??T,NKT and??T~+ NKT cells in NSCLC was significantly lower than that of normal controls.The relative counts of NKT,??T~+ NKT,??T~-NKT and CD~+_56??T cells in NSCLC was not sig- nificantly lower than that of normal control.The absolute counts of??T cells in NSCLC positively correlated to the number of NKT cells,(r=0.426,P=0.009).Conclusion The absolute counts of??T,NKT and their set ground cells in NSCLC was significantly lower than that of normal controls.In NSCLC the relation of the??T and NKT had positive correlation.

OBJECTIVE: To establish an HPLC method for the determination of Paraquat in biological samples. METHODS: Paraquat in biological samples was extracted by C18 columns which were pre-treated with cetyl-trimethyl ammonium bromide (CTAB) and soudium dodecyl sulphate (SDS), and analysed by HPLC/DAD. RESULTS: The detection limit of the method was 1 ng x mL(-1), and the average recoveries were 81%-94%. CONCLUSION The method can be used to analysis of paraquat in biological samples.
Animals ; Chromatography, High Pressure Liquid/methods* ; Herbicides/chemistry* ; Liver/chemistry* ; Paraquat/analysis* ; Rabbits ; Sensitivity and Specificity ; Sodium Dodecyl Sulfate ; Solvents ; Swine

Objective To investigate in vitro activity of antimicrobial agents against Enterococcus spp . isolated from clinic specimens in a hospital.Methods 188 Enterococcus spp . isolates from specimens sent by clinic depart-ments in June 2013-July 2014 were identified and performed antimicrobial susceptibility testing.Results Of 188 En-terococcus spp . isolates,119 were Enterococcus faecium (E.faecium),60 were E.faecalis ,and 9 were E.avium, these strains were mainly isolated from urine (34.57%)and blood specimens (19.15% ).No daptomycin and linezolid-resistant strain was detected;resistant rates of E.faecium to vancomycin was 1 .68%,to penicillin, ampicillin,high concentration gentamycin,erythromycin,and levofloxacin were all > 70%;except tetracycline, resistant rates of E.faecalis to the other antimicrobial agents were all lower than E.faecium,resistant rates of E. faecalis to penicillin and ampicillin were 16.67% and 13.33% respectively.Conclusion Daptomycin has high activity against Enterococcus spp . in this hospital.

Objective To investigate a method for the purification of the N?terminal peptide fragment(NT)of the myocardial calcium channel Cav1.2,and characterize its interaction with calmodulin(CaM). Methods EscherichiacoliBL?21 cells were transformed with plasmid pGEX?6p?3/NT harboring the NT?GST fusion gene. The cells harboring pGEX?6p?3/NT were cultured and protein expression was induced with isopropyl?β?D?thiogalactoside(IPTG). Then,the GST?NT fusion protein was purified by using glutathione Sepharose 4B(GS?4B)beads. GST was cleaved off with the PreScission protease,and SDS?PAGE was performed to detect the purity and relative molecular weight of the purified peptide. Further, GST pull?down assay was performed to characterize the interaction of the NT peptide with CaM. Results SDS?PAGE analysis showed that the NT peptide was successfully purified,with high purity. Results of the GST pull?down assay showed that the NT peptide could interact with CaM. Conclusion This study establishes a method for the purification of the NT peptide and lays the foundation for further research on the interaction partners and biological functions of NT.

OBJECTIVETo investigate the change of expression level of metastasis suppressor gene Kiss-1 in the colorectal cancer cell line SW480 after radiation, and to determine its association with the proliferation and apoptosis of SW480 cells.

METHODSSW480 cells were divided into control group (0 Gy) and study groups (2, 4, 6, 8 Gy). Cells in the study groups were irradiated by 6-MV X-ray radiation for 48 hours. Immunohistochemistry and real-time PCR methods were used to investigate the influence of radiation on Kiss-1 gene expression of SW480. Colony formation assay was used to detect the proliferation of SW480. Flow cytometry-Annexin- V/PI assay was used to observe the change of the apoptosis rate.

RESULTSCompared with the control group, Kiss-1 protein expression increased after radiation of 6, 8 Gy (P<0.05), but no significant changes were observed after radiation of 2, 4 Gy(P>0.05). Kiss-1 gene mRNA level increased after radiation of 2, 4, 6 Gy, while no obvious change was observed for 8 Gy radiation. The apoptosis rates increased for 4, 6, 8 Gy radiation(P<0.05), however, there was no significant difference for 2 Gy radiation (P<0.05).

CONCLUSIONRadiation may increase Kiss-1 gene expression in SW480 cells, which results in decreases proliferation and increases apoptosis in residual surviving cells.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; metabolism ; pathology ; Humans ; Kisspeptins ; genetics ; metabolism ; radiation effects ; RNA, Messenger ; genetics ; X-Rays

OBJECTIVETo study the gastric function after esophagectomy and cardiectomy with vagus nerve preserved and reconstruction of gastric funds (VPRG)in patients with esophageal cancer (EC) and cardiac cancer (CC).

METHODSSixty-eight patients with early or middle staged EC or CC received esophagectomy and cardiectomy with vagus nerve preserved and reconstruction of gastric funds (VPRG),while other 68 patients esophagectomy and cardiectomy with vagus nerve severed and no reconstruction of gastric funds (VSNG) as control. The symptoms,the pressure of the residual esophagus and thoracic stomach, 24-hour pH monitoring, mean basic gastric acid output, gastric emptying time of the intrathoracic stomach,fasting serum gastrin level, fibreoptic endoscopic results were compared before and after operation between the two groups.

RESULTSThe patients with VPRG had less symptoms after operation than those with VSNG such as anorexia, belch, reflux, heartburn, nausea, diarrhea, postcibal satiety (P< 0.01). In VPRG group,compared with the results before operation,there were no significant differences in 24-hour pH monitoring,the mean basic gastric acid output, the fasting serum gastrin level,the gastric emptying time of intrathoracic stomach one month and one year after operation (both P > 0.05). The pressure of the residual esophagus above the anastomosis in VPRG group was significantly higher than that in VSNG group (both P< 0.05). Fibreoptic endoscopic examination revealed higher incidences of postoperative atrophic gastritis and reflux esophagitis in VPRG group one month and one year after operation than those in VSNG group (P< 0.01).

CONCLUSIONPreservation of the vagus nerve and reconstruction of gastric funds after esophagectomy and cardiectomy for esophageal and cardiac cancer can prevent digestive disorder and improve the life quality of the patients.

Adult ; Esophageal Neoplasms ; surgery ; Esophagectomy ; methods ; Female ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Stomach ; physiopathology ; Vagus Nerve ; surgery

OBJECTIVETo explore the character of inorganic elements in Asari Radix et Rhizoma (Xixin).

METHODThe contents of 53 inorganic elements in Xixin samples from different localities and species were determined by ICP-AES and ICP-MS. The statistical data were made using SAS.

RESULTThe result demonstrated that Xixin has the high contents of Fe, Cr, Li. It has been observed that the content of Cu and Pb of the samples are much higher than the standard level. The results of hierarchical cluster analysis revealed two groups which correspond with the species of the samples. No correlations between the contents of the inorganic elements and the localities of the samples were found. Some characteristic elements were displayed in some specific areas. The difference of the contents of the 53 inorganic elements between root and rhizome of Xixin was reported for the first time. The primary form of inorganic elements in Xixin has been studied for the first time. The result demonstrated that the extraction rate between different elements varied, with the average extraction rate of (22.25 +/- 24.96)%.

CONCLUSIONThe inorganic elements analysis of Xixin can provide evidence of its identification, cultivation and application.

Asarum ; chemistry ; classification ; China ; Drugs, Chinese Herbal ; analysis ; Rhizome ; chemistry ; Trace Elements ; analysis

Objective Radionuclide-labeled low molecular weight polypeptide is reeently advocated for the diagnosis and treatment of malignant tumor. The purpose of this study was to evaluate the anti-tumor effect of 131Ⅰ-Tyr-octreotide in nude mice bearing human non-small cell lung cancer (NSCLC). Methods 131Ⅰ-Tyr-octreotide was prepared by Ch-T method. The radiochemical purity was measured and biodistribution was evaluated. The nude mice models bearing human NSCLC were studied and divided into four groups: group A injected 131Ⅰ-Tyr-octreotide through tail vein, group B injected normal saline, group C injected 131Ⅰ-Tyroctreotide through stroma and group D injected 131Ⅰ through stroma. The radioactivity ratio of tumor to normal tissue (T/NT) was calculated over region of interest (ROI). The tumor cell cycle and cell apoptosis were analyzed by flow cytometry (FCM), terminal deoxynucleotidyl transferase mediated dUTP-biotion nick end labeling (TUNEL) and histopathological analysis. Statistical analysis was performed with SPSS 11.0, and the comparison for difference between groups performed with one-way ANOVA analysis. Results The labeled radiochemical purity was (95.23±1.67)% and specific activity of 3.5×106Bq/ug. The biodistributiou showed high uptake in kidney, and low uptake in liver and spleen. The radioactive uptake in group C was higher than the other groups, and the retention time was longer. The T/NT was 52.74±0.13 after 24 h, which was much higher than that the other groups (group D: 8.90±0.23, group A: 6.42±0.02, q=628.81 and 664.33, all P<0.05). The resuits of tmnor cell cycle determined by FCM showed that the G1 phase was blocked mast remarkably in group C than the other groups [group C: (83.17±6.86)%, group A: (57.02±18.81)%, group D: (49.29±7.80)%, group B: (45.88±5.13)%, q=5.29, 6.86, 7.55, 1.56, 2.26, 0.69, all P<0.05]. Apeptotic cells were observed by TUNEL, and apoptotic body was detected by immuno-histochemical examination. Conclusions 131Ⅰ-Tyr-octreotide was easily labeled by Ch-T. 131Ⅰ-Tyr-octreotide could induce tumor cell apoptosis and inhibit the tumor cell of NSCLC. It might be a potential target-directed agent in NSCLC.

Objective To evaluate the cardio protective effect of astragaloside in rats with acute myocardial infarction and investigate the potential molecular mechanisms.Methods Forty-five Wistar rats were randomly divided into sham operation group,model group and astragaloside group,with 15 rats in each group.The acute myocardial infarction model was prepared by permanently occluding the left anterior descending artery in the model group and the astragaloside group,while the rats in the sham operation group were threaded only.Twenty-four hours after operation,the rats in the astragaloside group received astragaloside 20 mg · kg-1 · d-1 by intraperitoneal injection,once a day for two weeks.The rats in sham operation group and model group received isodose carboxymethylcellulose sodium by intraperitoneal injection,once a day for two weeks.Electrocardiogram was recorded to observe the changes of ST segment before ligation,after ligation immediately,1,3,7,14 d after intervened by astragaloside.Echocardiography was used to detect the left ventricular internal diameter at end-diastole (LVDD),left ventricular internal diameter at end-systole (LVDS),left ventricular end diastohc volume (LVEDV),left ventricular end systolic volume (LVESV),left ventricular fractional shortening (LVFS),left ventricular ejection fraction (LVEF).The levels of tumor necrosis factor alpha (TNF-α),interleukin (IL)-1 β,IL-6,nuclear factor-κB (NF-κB) in serum were measured by enzyme linked immunosorbent assay and the level of nitric oxide(NO) in plasma was measured by reagent kid.The expressions of toll-like receptor(TLR) 2,TLR4,nucleotide binding oligomerization do main(NOD) 1,NOD2,endothelial nitric oxide synthase (eNOS) mRNA were detected by quantitative real-time polymerase chain reaction.Results During the operation,there was 0,4,2 dead rats in the sham operation group,model group and astragaloside group.The ST segment in the model group and astragaloside group was higher than that in the sham operation group after operation immediately,1,3,7,14 d after intervented by astragaloside(P ＜ 0.01).The ST segment in the astragaloside group was lower than that in the model operation group at the time of 1,3,7,14 d after intervented by astragaloside(P ＜ 0.05).Compared with the sham operation group,the LVDD,LVDS,LVEDV,LVESV increased (P ＜ 0.01),LVFS,LVEF decreased (P ＜ 0.01),the levels of TNF-α,IL-1β,IL-6 and the expression of TLR2,TLR4,NOD1,NOD2 mRNA in cardiac muscle tissue were higher in the model group and astragaloside group (P ＜ 0.01),the level of NO was lower (P ＜ 0.05).The level of serum NF-κB in the model group was higher than that in the sham operation group(P ＜0.01),the expression of the eNOS mRNA in cardiac muscle tissue was lower than that in the sham operation group(P ＜ 0.05).There was no significant difference in the level of serum NF-κB and the expression of the eNOS mRNA in cardiac muscle tissue between the astragaloside group and sham operation group (P ＜ 0.05).Compared with the model group,in the astragaloside group the LVDD,LVDS,LVEDV,LVESV decreased (P ＜ 0.05),but LVFS,LVEF increased (P ＜ 0.05),the levels of TNF-o,IL-1 β,IL-6,NF-κB and the expression of TLR2,TLR4,NOD 1,NOD2 mRNA in cardiac muscle tissue were lower(P ＜ 0.05),the level of NO and the expression of the eNOS mRNA in cardiac muscle tissue were higher(P ＜ 0.05).Conclusion Astragaloside has protective effect against to acute myocardial infarction induced injury by decreasing the expression of TLR signal path and activating eNOS signaling.

BACKGROUND:Rapid and complete reperfusion has been widely adopted in the treatment of patients with acute myocardial infarction (AMI), but this process sometimes can cause severe reperfusion injury. This study aimed to investigate different patterns of post-conditioning in acute myocardial ischemia-reperfusion injury, and to detect the role of mitogen activated protein kinase (MAPK) during the injury. METHODS:Rats were randomly divided into five groups:sham group, reperfusion injury (R/I) group, gradually decreased reperfusion group (GDR group, 30/10-25/15-15/25-10/30 seconds of reperfusion/ischemia), equal reperfusion group (ER group, 20/20 seconds reperfusion/ischemia, 4 cycles), and gradually increased reperfusion group (GIR group, 10/30-15/25-25/15-30/10 seconds of reperfusion/ischemia). Acute myocardial infarction and ischemic post-conditioning models were established in the rats. Six hours after reperfusion, 3 rats from each group were sacrificed and myocardial tissues were taken to measure the expressions of phosphorylation of extracellular signal-regulated protein kinase (P-ERK), phosphorylated c-Jun N-terminal kinase (P-JNK), mitogen-activated protein kinase p38 (p38 MAPK), tumor necrosis factor-α (TNF-α), caspases-8 in the myocardial tissue, and cytochrome c in the cytosol using Western blot. Hemodynamics was measured at 24 hours after reperfusion, the blood was drawn for the determination of cardiac enzymes, and the heart tissue was collected for the measurement of apoptosis using TUNEL. One-way analysis of variance and the Q test were employed to determine differences in individual variables between the 5 groups. RESULTS:Three post-conditioning patterns were found to provide cardioprotection (P<0.05) compared with R/I without postconditioning. GIR provided the best cardioprotection effect, followed by ER and then GDR. Apoptotic index and serum marker levels were reduced more significantly in GIR than in ER (P<0.05). The enhanced cardioprotection provided by GIR was accompanied with significantly increased levels of P-ERK 1/2 (1.82±0.22 vs. 1.54±0.32,P<0.05), and lower levels of p-JNK, p38 MAPK, TNF-α, caspase-8, caspase-9 and cytochrome in the cytoplasm (P<0.05), compared with ER. The infarct size was smaller in the GIR group than in the ER group, but this difference was not significant (16.30％±5.22％ vs. 20.57％±6.32％,P<0.05). Allthe measured variables were improved more significantly in the GIR group than in the GDR group (P<0.05). CONCLUSION:Gradually increased reperfusion in post-conditioning could attenuate reperfusion injury more significantly than routine method, thereby the MAPK pathway plays an important role in this process.