Objective To evaluate the ability to kill human urothelial carcinoma T24 cells selectively in vitro by celecoxib combined with ZD55-IL-24 and explore the effectiveness for this combination use.Methods The EGFP expression of cells was observed by fluorescence microscope.The human umbilical vein endothelial cells (HUVEC) were used as crotrols.The expression of IL-24 mRNA was detected by RT-PCRwhen the cells were transfected by ZD55-EGFP or ZD55-IL-24.After transfected by ZD55-IL-24 and treated by celecoxib,the inhibition effect on cells was measured by MTT assay,and the apoptosis rate was examined by flow cytometry.Results The fluorescence in T24 and HUVEC can be observed 24h after ZD55-EGFP transfection and the fluorescence intensity was increased corresponding with the times.Fluorescence intensity in T24 cells showed higher than that in HUVEC group at the same times.The result of RT-PCR showed that the T24 cells expressed higher level of IL-24 mRNA than HUVEC group at the same time when the cells were transfect by ZD55-IL-24 (P ＜ 0.01).The inhibition rate of ZD55-IL-24 combined with celecoxib group was significantly higher than other groups (P ＜ 0.001).The inhibition rate of T24 cells in each group was significantly higher than HUVEC group (P ＜ 0.01).The flow cytometry results indicated that celecoxib combined with ZD55-IL-24 had the highest apoptosis rate on T24 cells than other single use group.Apoptosis rate of T24 cells showed a higher than HUVEC cells (P ＜ 0.01).Conclusion Celecoxib combined with ZD55-IL-24 can inhibit T24 cells proliferation at a greatest degree and this effect may be contributed to apoptosis.

Objective The study was aimed to detect the correlation of B cell activating factor (BAFF) promotor polymorphism-871 C/T and systemic lupus erythematosus (SLE).Methods BAFF promotor polymorphism-871 C/T was detected by the means of allele specific polymerase chain reaction (ASPCR) and agarose gel electrophoresis in 76 cases of SLE and 80 nonthrombosis normal individuals.The data of genotypic frequency and allele genotypic frequency were analyzed statistically with x2 test between the two groups.Results Fifty-one point two percent of normal individuals exhibited C/C.Thirty-five percent were heterozygous for C/T,and 13.8％ were homozygous for T/T.SLE group exhibited a different distribution pattern (30.3％ C/C,43.4％C/T,26.3％T/T).The allele frequency of T in SLE and normal individuals was 48.0％ and 31.2％ respectively.There was significant difference in the BAFF-871 C/T genotypic frequency between the SLE and nonthrombosis normal individuals (P＜0.05).Conclusion The polymorphism-871 C/T of BAFF promoter is correlated with the pathogenesis of SLE.The gene may be a major susceptible gene for SLE in Chinses Han people.Further investigations may be needed.

Phosphoinositide 3-kinase/serine-threonine kinase (PI3K/AKT)has been found playing an important role in the pathogenesis of severe acute pancreatitis (SAP)in recent years,but the underlying mechanism has not been clarified.Aims:To investigate the role of PI3K/AKT in regulating the inflammatory response in SAP by evaluating the effect of insulin-like growth factor-Ⅰ (IGF-Ⅰ)and wortmannin,the agonist and inhibitor of PI3K/AKT on Toll-like receptor 4 (TLR4)signaling pathway in macrophage cell line RAW264.7.Methods:RAW264.7 cells were treated with different concentrations of lipopolysaccharide (LPS ), IGF-Ⅰ and wortmannin, respectively, and cell viability was determined by CCK-8 assay.RAW264.7 cells were divided into blank control group (no treatment),LPS group (LPS 1 μg/mL),IGF-Ⅰ group (IGF-Ⅰ 100 ng/mL +LPS 1 μg/mL),wortmannin group (wortmannin 100 nmol/L +LPS 1 μg/mL)and IGF-Ⅰ +wortmannin group (wortmannin 100 nmol/L +IGF-Ⅰ 100 ng/mL +LPS 1 μg/mL).Protein expressions of tumor necrosis factor-α(TNF-α)and interleukin-6 (IL-6)were detected by ELISA;mRNA expressions of TLR4,myeloid differentiation factor 88 (MyD88),AKT,PI3K,p38 mitogen-activated protein kinase (p38MAPK)and nuclear factor-κB (NF-κB)were determined by real-time PCR.Results:After treated with LPS,IGF-Ⅰand wortmannin, respectively,no differences in cell viability of RAW264.7 cells were found between different concentrations groups (P>0.05).Protein expressions of TNF-αand IL-6 in LPS,IGF-Ⅰ,wortmannin and IGF-Ⅰ +wortmannin groups were significantly higher than those in blank control group (P<0.05 ).Protein expressions of TNF-αand IL-6 in wortmannin group were significantly lower than those in LPS and IGF-Ⅰ groups (P<0.05),and those in IGF-Ⅰ+wortmannin group were significantly lower than those in IGF-Ⅰ group (P<0.05).In LPS group,mRNA expressions of AKT and PI3K as well as TLR4 and its downstream molecules MyD88,p38MAPK and NF-κB were significantly higher than those in blank control group (P <0.05 ).Expressions of all above-mentioned mRNAs in IGF-Ⅰ group were further increased and significantly higher than those in LPS group (P<0.05).Expressions of all above-mentioned mRNAs in wortmannin group were significantly lower than those in LPS and IGF-Ⅰ groups (P<0.05 ),and those in IGF-Ⅰ+wortmannin group were significantly higher than those in wortmannin group (P<0.05),but significantly lower than those in IGF-Ⅰ group (P<0.05).Conclusions:PI3K/AKT might regulate TLR4 signaling pathway and its downstream molecules in macrophages, thereby affects the expressions of inflammatory cytokines and being involved in the pathogenesis of inflammatory response in SAP.

According to the Rules for the Implementation of the Standards for Review of Level 3 General Hospital (2011) of Formor Ministry of Health,the paper analyzes the requirements of review of level 3 general hospital for decision support,and takes the Sixth Affiliated Hospital of Sun Yat-sen University as an example to introduce the decision support solution and the application of Decision Support System (DSS).

Objective To explore the influence of integrated traditional Chinese medicine and western medicine on neurological function in patients with ischemic stroke.Methods One hundred patients with ischemic stroke were randomly divided into a treatment group and a control group.Both groups were given basic treatment.At this basis,sixty patients in the treatment group were given Chinese decoction,Kudiezi injection,traditional Chinese medicine soaking,and comprehensive rehabilitation; forty patients in the control group were given citicoline,modem rehabilitation techniques.Both groups were treated for 21 days.The scores of NIHSS,ADL,and MMSE were assessed at 7th day,14th day,and 21st day after admission of the trial,respectively.Results After 14 days,the scores of NIHSS (22.33± 1.06)in treatment group were decreased significantly (P＜0.05).After 21 days,the scores of NIHSS,ADL,MMSE in treatment group were(18.73±1.23),(20.19±2.17),(38.68±2.85),and(21.65±l.41),(25.37±2.34),(32.12±3.11)in control group,the improvement of scores in treatment group were superior to the control group (P＜0.05).The total effective rate was 91.67％ in treatment group,and 70％ in control group,it was significantly higher in treatment group than that in the control group (x2=7.98,P＜0.05).Conclusion The therapy of integrated traditional Chinese medicine and western medicine can significantly improve ischemic stroke,enhance the therapeutic efficiency of patients with ischemic stroke.

Objective Insulin autoantibody (IAA) is known to exist in sera of type 1 diabetes mellitus (T1DM) patients and pre-T1DM individuals. The aim of this study was to establish a novel microtiter plate radioimmunoassay (RIA) for IAA and evaluate its clinical value. Methods Diluted 125Ⅰ-insulin was mixed with 5 ul serum samples in a 96-well microtiter plate and then incubated for 72 h on an orbital plate shaker (4℃). The immunocomplexes were transferred to another protein a coated Millipore plate, and then the plate was washed with Tri-Buffered Saline Tween-20 (TBT) buffer. Counts per minute (CPM) was measured with liquid scintillation and luminescence counter. The positive cut-off point of IAA index was defined as ≥0.06 based on the 99-percentile of the distribution in 317 healthy individuals. The specificity and sensitivity of the assay were calculated from the samples provided by the fourth Diabetes Autoantibodies Standardization Program (DASP 2005). The IAA levels were determined in 71 T1 DM and 551 newly diagnosed type 2 diabetes (T2DM) patients, and 317 healthy controls. The t test, non-parametric test, x2 test and linear correlation analysis were performed on the data using SPSS 11.5 software. The concordance rate was estimated with Kappa value. Results (1) The optimized testing condition was described as 2×104 CPM of 125Ⅰ-insulin, 5 ul serum sample and slowly horizontal shaking for 72 h. (2) The intra-assay CV was 4.8%-8.9% and inter-assay CV was 6.4%-10.5%. Based on DASP 2005 samples, the specificity and sensitivity of the assay were 97% (97/100) and 50% (25/50), respectively. Ninety-six serum samples with different IAA levels were selected and tested to compare between our new method and a domestic IAA RIA kit. The results showed that the IAA indices from the two methods were positively correlated (r= 0.678, P<0.001). The concordance rate was 72.9 %(Kappa value=0.402). There were 25 samples with discordant results, which were positive for IAA titer using the corresponding microtiter plate RIA but negative using the novel RIA kit. (3) In TIDM group the positive rate of IAA was 19.7% (16/71), higher than the healthy controls (0.9%, x2=54.36, P<0.001). The subgroup of T1DM children (with 0-9 years) showed the highest IAA positive rate (55.6% ,x2=4.85, P<0.05). In T2DM group the frequency of IAA was 1.5% (8/551), which had no significant difference comparing with that of healthy controls (x2= 0.95, P >0.05). Conclusions Our proposed microtiter plate RIA method for IAA is highly sensitive and specific, likely to be feasible for clinical application. The frequency of IAA is high in children with T1DM.

Withanolide A is a biologically active secondary metabolite occuring in roots and leaves of Withania somnifera. In the present study, adventitious roots from leaf explants of W. somnifera were induced for the production of withanolide-A by Agrobacterium tumefaciens strain C58C1 to obtain hair roots. Hair roots induction rate reached 30%. The withanolide A was determined by HPLC in different hair roots lines and different parts of W. somnifera. The average content of withanolide A in all hair roots lines were 1.96 times as high as that in wild-plant, the concentration of withanolide A in hair roots (1.783 mg x g(-1) dry weight) were 1.51 times as high as the roots of wild W. somnifera (1.180 mg x g(-1) dry weight), respectively. It is possible to obtain withanolide A from hair roots culture of W. somnifera.
Agrobacterium tumefaciens ; physiology ; Plant Extracts ; analysis ; biosynthesis ; Plant Roots ; chemistry ; growth & development ; metabolism ; microbiology ; Withania ; chemistry ; growth & development ; metabolism ; microbiology ; Withanolides ; analysis ; metabolism

Objective To investigate the effect of granulocyte colony-stimulating factor(G-CSF)and its effect on the cognation in the PDGF hAPPV717I transgenic mice of Alzheimer's disease model.Methods Totally 36 PDGF hAPPV717I transgenic mice were randomly divided into two groups:G-CSF group and control group.The G CSF group was subcutaneously injected with 50 μg · kg-1 · d-1 of G-CSF for 7 days.The control group was injected subcutaneously with an equal volume of PBS in parallel.The animals were tested in Morris water maze on the 7th,14th,and 28th days after the last day of the injection,and the escape latency was recorded.Once the test was completed,the peripheral blood was taken to evaluate the effect of G-CSF to induce hematopoietic stem cells (HSCs) via flow cytometry.Then the mice were killed,their brains were quickly removed and frozen on dry ice.With the immunohistochemical staining and double immunofluorescence staining,the neurogenesis could be observed in the model mice.Results We found that G-CSF significantly shortened the escape latencies in PDGF-hAPPV717I transgenic mice compared to controls on the 7th,14th,and 28th day after G-CSF treatment [7 d:(27.19±4.07) s and (46.07±7.21) s,P＜0.000; 14 d:(26.48±5.29) sand (42.99±11.70) s,P＜0.010; 28 d:(24.97±3.61) s and (45.54±9.55) s,P＜0.002)].At the same time,we found that the percentage of CD34+/CD45+ cells in the peripheral blood was 0.358±0.161,0.223±0.038,0.168±0.049 on the 7th,14th,and 28th day after G-CSF treatment,respectively.Compared with the control group (0.073±0.026,0.067±0.034,0.072± 0.037),the percentage of CD34′ /CD45+ cells in the peripheral blood were increased (P＜0.001).BrdU+ cells were found in dentate gyrus (DG) of hippocampus and the cortex of the G-CSF group,where the BrdU+ /Ncstin- and BrdU-/MAP-2+ cells were also detected positively.Conclusions Subcutaneous administration of G- CSF may improve the cognition in APP transgenic mouse model of AD.G-CSF may mediate the proliferation,differentiation of hcmatopoietic stem cells (HSCs).and may induce the neural stem cells into the brain.

Objective To explore the influence of Danhong injection on neurological function and concentration level of high-sensitivity C-reactive protein (hs-CRP),tumor necrosis factor alpha(TNF-α),interleukin 6 (IL-6) in patients with acute cerebral infarction.Methods According to random number table method,eighty patients with acute cerebral infarction were randomly divided into a control group and a treatment gruop.Both groups were treated by aspirin,vitamine E,C and symptomatic treatment.At the same time,the treatment group was added Danhong injection,the control group was added low molecular dextran and citicoline,once a day,for 14 days.Results After the treatment,the concentration levels of hs-CRP [treatment group (2.11 ± 0.31) mg/L,control group (2.72 ± 0.23) mg/L],TNF-α [treatment group (0.35 ± 0.07) μg/L,control group (0.51 ± 0.06) μg/L],IL-6[treatment group (18.17 ± 3.15) pg/ml,control group (28.13 ± 4.97) pg/ml]was significantly lower than those before the treatment in both groups (P＜0.05).However,the concentration level of hs-CRP,TNF-α,IL-6 in treatment group was significantly lower than that in control group (P＜0.05).ADL scores [treatment group (20.26±2.17) scores,control group (28.83 ± 2.93) scores] and NIHSS scores [treatment group (18.34± 1.25) scores,control group s (22.52 ± 1.09) cores] were significantly lower after treatment than those before the treatment in both groups (P＜0.05),but all scores in treatment group were significantly lower than those in control group (P＜0.05).Conclusion Danhong injection decreased the concentration level of hs-CRP,TNF-α,IL-6 and NIHSS scores,ADL scores significantly.It also alleviated neurological function defect,enhanced the patient's activities of daily living,and reduced the disability rate of ischemic brain damage.

BACKGROUND: Human umbilical cord mesenchymal stem cells (HUMSCs) can differentiate into insulin-producing cellsafter induced by chemical drugs or co-culture methods, but insulin secretion is extremely low. Therefore, to inducemature pancreatic beta cell differentiation from stem cells by adenovirus transfection of specific genes involved in thedevelopment of pancreas is a research hotspot in recent years.OBJECTIVE: To study the differentiation potential of HUMSCs into insulin-producing cells after transfection withmusculoaponeurotic fibrosarcoma oncogene homologue A (MafA).METHODS: Ad-MafA-EGFP was transferred into passage 3 HUMSCs. After 7 days of induction, changes of cellmorphology were observed by inverted phase contrast microscope. Expression of pancreatic cell-specific genes(glucagon, PDX1, Nkx2.2) was detected by PCR technique.RESULTS AND CONCLUSION: After Ad-MafA-EGFP transfection, no significant morphological changes were observedin the HUMSCs under inverted phase contrast microscope. It was confirmed by fluorescence microscope thatAd-MafA-EGFP was transferred into the HUMSCs. After induction, the expression of human pancreatic precursorcell-related genes, including glucagon, PDX1 and Nkx2.2, was increased as detected by PCR. To conclude, thesefindings could provide experimental evidence for further differentiation and maturation of pancreatic β cells fromHUMSCs.