Objective: To investigate G-CSF expression and rhG-CSF's effects on cell proliferation and its mechanism in human solid carcinoma cell lines. Methods: G-CSF expression was detected by human G-CSF ELISA Kit. Cell proliferation promoted by rhG-CSF was studied by cell count and MTT. The influence of rhG-CSF on KB cell cycle and apoptosis was analysed by flow cytometry. G-CSF receptor expression was detected by immunocytochemistry. Results: Cell line T-24 produced large amount of G-CSF; rhG-CSF promoted cell line KB proliferation in a dose-dependent manner and reached its maximal effect at a concentration of 100 ng/mL (P < 0.05); rhG-CSF antagonized G0/G1 phase block and reduced apoptosis rate of KB cells. Conclusion: Cell line T-24 expresses G-CSF; rhG-CSF promotes, KB cell proliferation by binding its G-CSF receptor, antagonizing G0/ G1 phase block and inhibiting apoptosis.

Objective To investigate the set-up errors of super and middle part of esophageal cancer patients using cone-beam CT (CBCT) during intensity modulated radiotherapy (IMRT),hence determine various margins from CTV to PTV.The corresponding influence on the normal tissues (lung and spinal cord) was also discussed.Methods From December 2012 to December 2013,12 patients with upper and middle segment of esophageal cancer were chosen.Using their 60 sets of weekly acquired CBCT images prior to the treatment,the lateral,longitudinal,and vertical set-up errors of each patient were obtained.Based on these measured errors and the target motions,we adopted new margins to create new PTV.Then IMRT plans were created for the original PTV (5 mm margin in all directions on CTV) and new PTV respectively.On condition of the same target coverage (V95 ≥ 95％),the doses to lungs (V5,V20,V30,D) and spinal cord (D1 cm3) were compared statistically between the original and new plans.Results According to the 60 CBCT scans,the average left-right (RL),superior-inferior (SI),anterior-posterior (AP) set-up errors were (2.02 ± 1.74),(2.03 ± 1.93),and (2.02 ± 1.89) mm respectively.The margins were 5.6 mm (RL),8.5 mm (SI),and 4.7 mm (AP) for the upper esophagus and 6.2 mm (RL),11 mm (SI),and 5.0 mm (AP) for the middle esophagus.Comparison of both lungs and spinal cord suggested significant differences between the two plans (t =-8.23,-5.55,-4.66,-6.87,-4.67,P ＜0.05).Conclusions The margins from CTV to PTV should be created via CBCT-measured set-up errors and previous reports,which can be helpful for clinical treatment.

Objective To investigate the damage mechanism of fluetuant high glucose on the INS-1 cells (pancreatic β-cell lines).Methods The cells were divided into five groups:the control groups (A group:5.5 mmol/L of glucose),the continuing high glucose group (B group:16.7 mmoL/L of glucose),the fluctuant glucose group ( C group:16.7 mmol/L of glucose for cultivation for 2 h,then the concentration changed to 5.5 mmol/L for cultivation for 3 h,which was repeated 3 times per day;the ceils were kept in the medium containing 5.5 mmol/L of glucose during night time for 9 h),the continuing high glucose plus NAC ( 1.0 mmo/L) group ( D group),the fluctuant glucose plus NAC group ( E group).The intracellular reactive oxygen species (ROS) was observed by the flow cytometry.The activity of glucose-6-phosphate dehydrogenase (G6PD) was estimated by the tetrazolium linked cytochemical method.Results 72 h after intervention,the levels of ROSwere 37.77±2.31,86.97±7.97,124.27±10.04,60.92±2.61 and 51.47±3.36,respectively,in A～E group;the activities of G6PD were 1.25±0.03,1.09±0.02,1.03±0.01,1.12±0.02 and 1.21±0.01,respectively;the levels of NADPH were (0.123±0.003) mmol/mg prot,(0.112±0.004) mmoL/mg prot,(0.099±0.002 ) mmol/mg prot,( 0.116±0.005 ) mmol/mg prot and ( 0.120±0.002) mmol/mg prot,respectively.The level of ROS in the cells of the fluctuant glucose group were significantly higher than that in the continuing high glucose group ( P ＜ 0.01 ).The G6PD activity and NADPH was significantly lower in fluctuant high glucose group than those in the continuing high glucose group (P ＜0.01 ).NAC co-cultivation decreased the extent of cell's change.Conclusions Exposure of INS-1 to high glucose lead to increased oxidative stress, possible mechanism included decreased G6PD activity and subsequent imbalance between oxidation and reduction.

Objective To clone and express prokaryotic recombinant plasmid of nucleoside triphosphate hydrolase (NTPase) gene of Toxoplasma gondii, and analyze its antigenicity. Method NTPase gene was amplified by PCR from RH strain of T. gondii and cloned into pGEM-T Easy vector. Positive clones were screened and identified by BglⅡ, HindⅢ digestion and sequenced. The target gene was then subcloned into prokaryotic expression vector pBAD-HisB and transformed into E. coli BL21(DE3). The expressed recombinant protein was purified with Ni-NTA agarose and further analyzed by sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and Western blotting. Results NTPase-Ⅱ gene was specifically amplified, and the homology of DNA sequence was 100% to that in the GenBank. SDS-PAGE showed that the recombinant NTPase protein with correct molecular weight was expressed highly in E.coli BL21(DE3). Western blotting testified that the purified recombinant protein could be specifically recognized by mouse serum immunized with T. gondii and mouse anti-recombinant protein serum. Conclusion The NTPase-Ⅱ gene has been cloned and expressed in E.coli BL21(DE3), and the purified protein of NTPase-Ⅱ gene displays a specific antigenicity.

OBJECTIVE To establish a simpler method for screening bacterial metallo-?-lactamase(MBL) by comparing Hodge test and double-disk synergy tests(DDSTs),and using 5 inhibitors such as sodium citrate,EDTANa2,EDTAK2,sodium dimercaptopropane sulfonate and mercaptoethanol.METHODS In Hodge test,5 inhibitors of different concentration were added to the imipenem(IMP) disks,the sizes of the inhibition zones were compared.Disk containing IMP and disk containing a metallo-?-lactamase inhibitor were used in DDSTs.The optimal inhibitors concentration and edge-to-edge distance between the two disks were selected.The performances of the Hodge test and the DDSTs were also compared.RESULTS Among the metallo-?-lactamase inhibitors used in this study,1∶15 diluted mercaptoethanol gave the most reproducible and the clearest results when a filter disk containing mercaptoethanol was placed near the IMP disk with the edge distance of 3 mm.CONCLUSIONS Mercaptoethanol-IMP DDSTs are simple,convenient and sensitive methods for screening MBL.

We analyzed the pathogenic characteristics of Salmonella strains isolated from diarrhea patients in Wuxi City,Jiangsu Province,China and compared the differences among pulse field gel electrophoresis (PFGE) patterns of main serotype strains,so as to provid scientific basis for disease control.After biochemical identification of the Salmonella strains isolated from infectious diarrhea patients in Wuxi in 2015,drug susceptibility test,serotyping and PFGE were applied to analyze these strains.Results showed that a total of 32 Salmonella strains were detected from 756 diarrhea specimens with a positive rate of 4.23 ％.The infection occurred more frequently between May and October and adults aged more than 60 years old affected mostly.There was no significant difference between genders in infected population.The drug susceptibility test indicated that the antibiotic resistance rate of these Salmonella strains to ampicillin (56.25 ％) was the highest,and to ciprofloxacin(6.25 ％)and Ceftazidime (6.25％) were the lowest.The 32 Salmonella strains belonged to 11 serotypes,and S.enteritidis(31.25％)and S.typhimurium(21.88％) were the predominant serotypes.PFGE showed that the pattern similarity of all S.enteritidis was more than 85 ％;PFGE patterns of S.typhimurium were different.In conclusion,the infection of Salmonella from diarrhea patients in Wuxi City had obvious season and age specific distribution,and the most prevalent serotype of Salmonella was the S.enteritidis.It is necessary to strengthen the surveillance of Salmonella concurrently in food and environment.

ObjectiveTo evaluate setup errors of image guided radiation therapy (IGRT) for body tumors immbilized with vacuum cushions and localized with final isocenter marked method using kilovoltage cone beam CT (KVCBCT).MethodsA retrospective study has been carried out for 223 patients from March 2009 to April 2011.All patients were immobilized with vacuum cushions and localized with final isocenter marked method in Philips PQS CT or Philips Brilliance CT Big Bore scanner,which were equipped with LAP movable laser systems. The CT images were transferred to a Varian Eclipse workstation for contouring and treatment planning.Before irradiation,a KVCBCT scan was performed and image registration was done on a Varian iX linear accelerator via OBI system.Each set of setup errors in right-left ( RL),superior-inferior (SI),and anterior-posterior (AP) directions were gathered.An independent-samples t-test statistical analysis was conducted with the 758 sets of data using SPSS 16.0.Results The statistical analysis showed that setup errors from the 758 datasets were depicted a Gaussian distribution.The system errors ± random errors in RL,SI and AP were ( -0.5 ±2.8) mm,(0.0±3.0) mm and (0.4±3.4)mm,respectively.Referring to the formula for planning target volume margin calculation,M =2.5Σ + 0.7δ,the margins were calculated as 3.2,2.1 and 3.4 mm,respectively.ConclusionsThe margins derived from this retrospective study have confirmed the premise that the treatment plans were executed in patients with high reliability,thereby created a high sense of confidence for the clinicians.

Objective To investigate the relationship between the steroid-sensitive nephrotic syndrome(SSNS) and interleukin-18(IL-18) and to approach the inhibitive role of dexamethasone(DEX) on expression of IL-18 of peripheral blood mononuclear cells(PBMC) in children with SSNS in vitro.Methods IL-18 levels of serum, urine and supernatants of PBMC cultured in vitro were measured by enzyme linked immunosorbent assay(ELISA) in 23 children with SSNS who were either before or after treatment. Fifteen age-matched healthy children served as normal control group, and another 18 children with respiratory infections as infectious control group.Results There were signi-ficant differences of IL-18 in serum and urine before and after treatment in children with SSNS (t=15.072,16.149 Pa

BACKGROUND: The virulent factors ofEscherichia coli (E.coli) play an important role in the process of pathopoiesis. The study aimed to compare drug-resistant genes and virulence genes between extended spectrum β-lactamases (ESBLs)-producingE.coli and non-ESBLs-producing E.coli to provide a reference for physicians in management of hospital infection. METHODS: From October 2010 to August 2011, 96 drug-resistant strains ofE.coli isolated were colected from the specimens in Qingdao Municipal Hospital, Qingdao, China. These bacteria strains were divided into a ESBLs-producing group and a non-ESBLs-producing group. Drug sensitivity tests were performed using the Kirby-Bauer (K-B) method. Disinfectant gene, qacEΔ1-sull and 8 virulence genes (CNF2, hlyA, eaeA, VT1, est, bfpA, elt, and CNF1) were tested by polymerase chain reaction (PCR). RESULTS: Among the 96E.coli isolates, the ESBLs-producingE.coli comprised 46 (47.9％) strains and the non-ESBLs-producingE.coli consisted of 50 (52.1％) strains. The detection rates of multiple drug-resistant strain, qacEΔ1-sull, CNF2, hlyA, eaeA,VT1, est, bfpA, elt, and CNF1 in 46 ESBLs-producingE.coli isolates were 89.1％, 76.1％, 6.5％, 69.6％, 69.6％, 89.1％, 10.9％, 26.1％, 8.7％, and 19.6％, respectively. In the non-ESBLs-producingE.coli strains, the positive rates of multiple drug-resistant strain, qacEΔ1-sull, CNF2, hlyA, eaeA, VT1, est, bfpA, elt, and CNF1 were 62.0％, 80.0％, 16.0％, 28.0％, 64.0％, 38.0％, 6.0％, 34.0％, 10.0％, and 24.0％, respectively. The difference in the detection rates of multiple drug-resistant strain, hlyA and VT1 between the ESBLs-producingE.coli strains and the non-ESBLs-producingE.coli strains was statistically significant (P<0.05). CONCLUSION: The positive rate of multiple drug-resistant strains is higher in the ESBLs-producing strains than in the non-ESBLs-producing strains. The expression of some virulence genes hlyA and VT1 varies between the ESBLs-producing strains and the non-ESBLs-producing strains. Increased awareness of clinicians and enhanced testing by laboratories are required to reduce treatment failures and prevent the spread of multiple drug-resistant strains.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Liver ; pathology ; Liver Cirrhosis ; complications ; pathology ; Liver Neoplasms ; etiology ; pathology ; Male ; Middle Aged ; Young Adult