Objective To evaluate the selectively killing effect of adenovirus(Ad)mediated double suicide gene driven by VEGF promoter on pancreatic cancer cell SW1990. Methods VEGFexpressing SW1990 were infected by Ad-VEGFP-CDTK and Ad-null.respectively.The infection rate was observed and the expression of CDTK was detected by RT-PCR and Western blotting.Followed by treatment with 5-FC and GCV killing effects were evaluated and bystander effects were analyzed by MTF.Pathological character of cells was observed by electron microscopy and distribution of cell cycle was detected by flow cytometry.The caspase-3 activity was detected by absorption spectrometry. Results The infection rate of the resultant recombinant Ad to SW1990 cells was not apparently different.RT-PCR and Western blotting demonstrated product of CDTK gene in SW1990 cell infected by Ad-VEGFP-CDTK.Prodrug could inhibit proliferation of SW1990 and the effect was dose-dependent.There was considerable bystander effect as observed by MTF.Apoptotic peak was also shown by flow cytometry.Morphologic features of apoptosis in SW1990 cells were displayed via electron microscopy.Cells at the G0-G1 phase was increasing and the rate at the G2-M and S phase was decreased by prodrug.The caspase-3 activity gradually rised with the increasing concentration of the prodrug. Conclusions The CDTK fusion gene system controlled by VEGF promoter has killing effect on the VEGF-expressing SW1990 cells and inducing the cell apoptosis.

Objective To investigate the diagnostic value of anti-mutated citrullinated vimentin (anti-MCV) and anti-cyclic citrullinated peptide (anti-CCP) antibodies for ankylosing spondylitis (AS), and to establish a new diagnostic method of AS based on anti-citrullinated protein/peptide antibodies (ACPAs) detection.Methods Totally 121 AS patients(AS group), 97 rheumatoid arthritis(RA) patients (RA group)and 103 healthy people(control group) were enrolled for the detection of serum levels of anti-MCV and anti-CCP antibodies using ELISA method as a diagnostic test .HLA-B27 in AS patients was tested by flow cytometry , and RF-IgM in RA and AS patients was detected by immune rate nephelometry .Receiver operating characteristics ( ROC) curve was used to analyze the diagnostic value and to determine the cut-off value.Anti-MCV and anti-CCP antibodies among each group were compared by Mann-Whitney U test, Chi-square test or Fisher′s exact test was used to compare positive rate .Results The anti-MCV levels in AS group [11.60 ( 12.36-25.83 ) U/ml ] were significantly higher than control group [ 11.60 ( 8.41 -13.54)U/ml ],U=2 413,P <0.001, while the levels of anti-CCP had no difference between the two groups [AS group 6.22 (4.30 -8.07) U/ml], and control group [6.01 (3.77 -7.58) U/ml], respectively;U=5 421,P=0.094.The calculated area under the ROC curve of anti-MCV was 0.806,and 14.67 U/ml was the optimal cut-off value with sensitivity of 0.645 and specificity of 0.942.In both HLA-B27 positive and negative AS patients , anti-MCV antibodies levels and positive rate were significantly higher than control group using new cut-off value above (U =133.5, P =0.001; U =2 279.5,P <0.001). Sensitivity of the combination detection of anti-MCV and anti-CCP ( MCV+CCP-) or RF-IgM ( MCV+RF-) were 60.3%(73/121), 62.8% (76/121) and specificity were 89.7% (87/97), 90.7% (88/97) respectively in differential diagnosis of AS and RA , which were significantly higher than anti-MCV detection alone in specificity (16.5%,16/97).Conclusions Anti-MCV could be a new biomarker for the diagnosis of AS .With high sensitivity and specificity , anti-MCV has an equal diagnostic efficiency in HLA-B27 positive and negative AS patients using our new cut-off value.Additionally, the combination detection of anti-MCV and anti-CCP or RF-IgM could be an effective method for differential diagnosis of AS and RA .

Objective To investigate the curative effect of the adenovirus-mediated fusion gene system driven by KDR promoter (AdKDR-CDglyTK) on a model of pancreatic cancer. Methods By using transplantation of the cultivated cells, human pancreatic cell line Capan-2 was injected subcutaneously on the back of nude mice to establish the animal model of the pancreatic cancer. Twenty nude mice were divided randomly and equally into four groups. The mice in group Ⅰ were injected with AdKDR-CDglyTK and 5-FC/GCV, those in group Ⅱ were injected with 5-FC/GCV, those in group Ⅲwere injected with AdKDR-CDglyTK and those in group Ⅳ received no any injection. AdKDR-CDglyTK was injected directly into the tumor and 5-FC/GCV was given by intraperitoneal injection. The observing parameters included common status, tumor bulk, tumor weight, inhibition rate of tumor growth, pathology, immunohistochemistry and treatment effect in each group. Electron microscopy was performed to observe the pathological changes of cells. The apoptotic cells in tumor were detected using the TUNEL assay. The expression of CDglyTK in tumors from each group was examined by RT-PCR. Results Tumor growth was dramatically inhibited in group Ⅰ. Tumor growth has no significant difference among groupⅡ , group Ⅲ and group Ⅳ. The apoptotic rate (34.20±4.60)% was significantly increased in group Ⅰ (F= 243. 22, P= 0. 00) and it had no significant difference among groupⅡ , group Ⅲ and group Ⅳ (P＞0.05). Conclusion AdKDR-CDglyTK with 5-FC/GCV can obviously inhibit the growth of human KDR-expressing pancreatic cell line Capan-2 and induce the cell apoptosis in vivo. The probable molecular mechanism lies in the facts that the system can cause a decline in the level of Bcl-2.

Objective Radiofrequency ablation (RFA) and endoscopic breast conserving surgery (EBCS) has been developed as a potential means for treating breast tumors with minimal disruption to adjacent soft tissues.The purpose of this study was to evaluate the cosmetic outcomes of RFA assisted by EBCS on patients with early breast carcinoma (T1 -2 N0-1 M0).Methods Twenty-three patients with biopsy proven invasive breast carcinoma ( ≤3 cm in diameter),were treated with ultrasound (US)guided percutaneous-RFA for the local tumors followed by sentinel lymph node biopsy and endoscopic auxiliary node dissection and lumpectomy.The reactions to breast cosmetics were observed and compared to the conventional approaches using the breast asymmetry score for EBCS and a self -assessment questionnaire targeting objective and subjective data about treatment related breast change.Results Twenty-three patients were successfully treated with RFA.Complete coagulation necrosis of the tumor was visualized at US and mammography in 22 of 23 patients (95.6 ％).22 patients being per formed RFA and EBCS presented little scars and the breasts preserved more cosmetically acceptable than the reported results of traditional protocols.Self-assessment acceptable rate was 100 ％,aesthetic outcome as good or excellent rated in 95.5 ％(21/22).Within the follow-up of above 36 months,there were no local recurrence,metastasis,wound infection or necrosis,nerve damage,haematoma,as well as upper limb swelling or lymphoedema,no reduced upper limb mobility and tumor seeding.Conclusions RFA with endoscopic breast conservation therapy for the treatment of early breast carcinoma is more effective and cosmetically acceptable than the traditional approaches.

Objective To observe the clinical effect of hypomethylating agents(HMAs)in the treatment of patients with intermediate-and high-risk myelodysplastic syndrome(MDS).Methods A retrospective study was conducted in 58 patients with intermediate-and high-risk MDS.The study group(25 patients)received azacitidine or decitabine for hypomethylating treatment,while the control group(33 patients)received routine symptomatic supportive treatment.The clinical efficacy,hemato-logic parameters,quality of life,and adverse events were observed and compared between the two groups.Results After treatment,the complete remission rate,objective response rate,and disease control rate were higher in the study group than in the control group,while the disease progression rate was lower(P＜0.05).The 1-year survival rate was 92.00％in the study group,which was higher than 75.76％in the control group,but the difference showed no statistically significant(P＞0.05).After treatment,hemoglobin,white blood cell,and platelet levels were higher in both groups than before treatment,and were higher in the study group than in the control group(P＜0.05).The total incidence of adverse events was lower in the study group than that in the control group(P＜0.05).After treatment,the quality of life scores(physical function,cognitive function,emotional function,role function,and social function)were higher in the study group than those in the control group(P＜0.05).Conclusion HMAs treatment is superior to routine symptomatic supportive treat-ment in patients with intermediate-and high-risk MDS,and not only improves the quality of life but al-so has good safety.

Objective To observe the clinical effect of hypomethylating agents(HMAs)in the treatment of patients with intermediate-and high-risk myelodysplastic syndrome(MDS).Methods A retrospective study was conducted in 58 patients with intermediate-and high-risk MDS.The study group(25 patients)received azacitidine or decitabine for hypomethylating treatment,while the control group(33 patients)received routine symptomatic supportive treatment.The clinical efficacy,hemato-logic parameters,quality of life,and adverse events were observed and compared between the two groups.Results After treatment,the complete remission rate,objective response rate,and disease control rate were higher in the study group than in the control group,while the disease progression rate was lower(P＜0.05).The 1-year survival rate was 92.00％in the study group,which was higher than 75.76％in the control group,but the difference showed no statistically significant(P＞0.05).After treatment,hemoglobin,white blood cell,and platelet levels were higher in both groups than before treatment,and were higher in the study group than in the control group(P＜0.05).The total incidence of adverse events was lower in the study group than that in the control group(P＜0.05).After treatment,the quality of life scores(physical function,cognitive function,emotional function,role function,and social function)were higher in the study group than those in the control group(P＜0.05).Conclusion HMAs treatment is superior to routine symptomatic supportive treat-ment in patients with intermediate-and high-risk MDS,and not only improves the quality of life but al-so has good safety.

AIM:The study aimed to explore the effect of curcumin(Cur)on lipopolysaccharide(LPS)-in-duced RAW264.7 cell-derived osteoclasts,together with its underlying mechanism.METHODS:An osteoclast model was established by treating RAW264.7 cells with LPS.The viability of the cells was assessed by CCK-8 assays and osteo-clast formation was evaluated by tartrate-resistant acid phosphatase(TRAP)activity.The levels of reactive oxygen species(ROS),Fe2+,glutathione(GSH),and malondialdehyde(MDA)were examined by biochemical assays.Mitochondrial morphology was assessed by transmission electron microscopy.The mRNA and protein levels of p53,glutathione peroxi-dase 4(GPX4),and solute carrier family 7 member 11(SLC7A11)were determined by RT-qPCR and Western blot,re-spectively.RESULTS:Treatment with LPS successfully induced osteoclasts formation in RAW264.7 cells.The TRAP results showed that compared with the LPS-treated group,the number of osteoclasts and TRAP activity in the curcumin-treated group decreased dose-dependently(P<0.01).Compared with the control group,the LPS+Erastin group showed significantly increased TRAP activity(P<0.01),while after curcumin treatment,the TRAP activity declined in a dose-de-pendent manner(P<0.01).The results of the biochemical tests showed that compared with the control group,the LPS+Erastin group had significantly elevated levels of ROS,Fe2+,and MDA,while the GSH level was significantly reduced(P<0.01),and compared with the LPS+Erastin group,the ROS,Fe2+,and MDA levels in the curcumin group decreased(P<0.01)and GSH levels increased(P<0.01).These effects were all dose-dependent.Transmission electron microscopy showed that compared with the LPS group,the LPS+Erastin group had reduced mitochondrial cristae and increased mem-brane density,while after treatment with curcumin,both these effects were reversed.The RT-qPCR and Western blot re-sults showed that compared with the control group,the mRNA and protein levels of p53 in the LPS+Erastin group were sig-nificantly increased,while those of of SLC7A11 and GPX4 were significantly reduced(P<0.01).After curcumin treat-ment,the p53 mRNA and protein levels were reduced while the levels of SLC7A11 and GPX4 were increased(P<0.01).CONCLUSION:Curcumin can inhibit lipopolysaccharide-induced differentiation of RAW264.7 cells into osteoclasts,and its mechanism may be related to the inhibition of the p53/SLC7A11/GPX4 signaling pathway.