Objective To construct a luciferase gene expression vector containing full-length 3' untranslated region(3'UTR)of mouse vascular endothelial growth factor-C(VEGF-C)gene,and to observe the effects of VEGF-C 3'UTR on luciferase gene expression by a double-fluorescence report system.Methods Polymerase chain reaction(PCR)was used to amplify VEGF-C 3'UTR and a 312bp VEGF-C coding region(CR)fragment from full-length VEGF-C cDNA in mouse Lewis lung cancer cells.The luciferase expression vectors containing VEGF-C 3'UTR or VEGF-C CR were constructed by subcloning the PCR products to luciferase reporter vector pGL3-Promoter using gene engineering technology,and then they were transfected to mouse Lewis lung carcinoma cells by LipofectamineTM 2000.The activities and mRNA expression of luciferase were detected by Dual-Luciferase Reporter System and quantitative RT-PCR,respectively.Results Mouse VEGF-C 3'UTR(429bp)and VEGF-C CR(312bp)were successfully amplified by PCR.The VEGF-C 3'UTR and VEGF-C CR fragments were successfully inserted into pTA2 vector,and then subcloned to pGL3-Promoter vector at Xba Ⅰ site by using restriction endonucleases analysis.The DNA sequences and insertion orientation of PCR products were all correct by sequencing analysis.The resulted luciferase expression plasmids were named pGL3-VEGF-C 3'UTR and pGL3-VEGF-C CR,respectively.Dual-Luciferase Reporter System detection and quantitative RT-PCR showed that in Lewis lung carcinoma cells,the activities of luciferase and expression of luciferase mRNA in the pGL3-Promoter group were higher than that in the pGL3-VEGF-C 3'UTR group,and there was no significant difference between pGL3-VEGF-C CR group and pGL3-Promoter group.Conclusion VEGF-C 3'UTR can inhibit luciferase gene expression.

Objective To explore whether alternatively activated macrophages (aaMphi) can transdifferentiate into lymphatic endothelial cells (LEC) under the inducement of VEGF-C, and to investigate the possible mechanisms involved in aaMphi-induced lymphangiogenesis. Methods An aaMphi model constructed by treating mouse macrophage cells RAW264.7 with mouse recombinant IL-4 for 24h was treated with different concentrations of recombinant mouse vascular endothelial growth factor-C (VEGF-C). After the clustering formed and the tube-like structures were detected in the matrigel, the aaMphi transdifferentiation system was finally decided to be constructed with the VEGF-C in the concentration of 100ng/ml. The mRNA expression of LEC specific markers, VEGFR-3 and Prox1, and aaMphi specific marker, Fizz1, were detected by real time quantitative RT-PCR on the 0, 7th, 14th, and 28th day, respectively, after VEGF-C stimulation. Formation of tube-like structure in the matrigel was observed with inverted phase contrast microscope in 28 consecutive days. Results The VEGF-C induced transdifferention system, incubated in EBM-2 medium and sustained by the matrigel, was successfully established. In this system, it was found that the mRNA expression of VEGFR-3 and Prox1 gradually increased, whilst that of Fizz1 decreased. The mRNA expression of VEGFR-3 and Prox1 reached the peak value, whilst that of the Fizz1 went down to the nadir, on the 14th day. No significant difference in values was found between the 14th day and the 28th day. During the period of the 7th day to the 28th day, distinct tube-like structures were gradually formed in the matrigel and the numbers increased in a time-dependant manner. Conclusion VEGF-C can induce the transdifferentiation of aaMphi into LEC by up-regulating the mRNA expression of VEGFR-3 and Prox1 in aaMphi, which is one of the possible mechanisms involved in aaMphi-induced lymphangiogenesis.

Objective:To investigate the effects of antimalarial artemether on the proliferation of human lung adenocarcinoma cell line A549 in vitro and provide experimental data for the treatment of lung cancer with artemether.Methods:MTT assay was used to observe the inhibitory effects of artemether on the proliferation of A549 cells.Cell growth curve was draw according to the cell counts.The population doubling time was obtained in logarithmic growth phase.Cell cycle detection was observed by flow cytometry.H-E staining and transmission electron microscopy were used to observe the altered morphology of apoptotic cells. Results:Artemether has a significantly inhibitory effect on the proliferation of A549 cells in a dose-dependent manner in vitro,and the IC50 was 1.34 mg/L.The population doubling time in logarithmic growth phase in the artemether treatment group was(20.7?0.5) h compared to(32.2?0.3) h in the control group.The difference between two groups was statistically significant(P

Objective To explore the role of chemokine CXCL12 and its receptor CXCR4 in the directional migration of human prostate cancer(PCa).Methods The expression of CXCL12/CXCR4 in 18 human PCa samples and human PCa cell lines(PC3,DU145 and LNCap)was determined by immunohistochemistry and immunocytochemistry,respectively.Then the effect of CXCL12 on the migration and invasion of human PCa cell lines Was investigated by Matrigel invasion assay.Results Except 1 PCa sample,positive CXCR4 protein expression was detected in 17 clinical PCa samples.On the contrary,in 18 samples determined,only one sample expressed weak CXCL12 protein.CXCR4 rather than CXCL12 protein was exressed in PCa cell lines PC3,DU145 and LNCap.In addition,CXCL12 promoted the migration and invasion of PCa cell lines in a dose dependent manner in viiro,in which experiments PC3,LNCap cells were pretreated by antibody of CXCL12 or CXCR4 and then it was found the migrations of cells stimulated by CXCL12 were inhibited.Conclusion CXCR4 protein is expressed in human PCa and CXCL12/CXCR4 axis may play a significant role in the metastasis of prostate cancer.

The X-ray computed tomography (CT) appearance of 4 cases with pulmonary cryptococcosis (PC) diagnosed by pathological examination in our hospital was retrospectively analyzed. The appearances of PC on CT were various: solitary lesion in 1 case, multiple lesions in single lobe in 2, and multiple lesions in multiple lobes in 1. There were total 52 lesions in 4 cases; the diameter of nodules or masses was 3 - 75 mm. Cavitations were found in 1 case; lesions appeared obviously enhanced and one lesion showed central necrosis. Two cases underwent pulmonary lobectomy; and 2 cases received core cutting needle biopsies, after antifungal therapy for 3 months to 1 year the lesions showed being absorbed. In summary, the CT appearance of PC is non-specific with various modes and forms. PC should be considered when multiple nodules or masses scattered in subpleural zone, accompanied with ground-glass opacity and obviously enhanced. The examination of pathogen and pathology at the beginning is crucial for improving diagnostic accuracy.

AIM: To investigate the effect of glomerular intercellular interaction under high glucose concentration on the production of reactive oxygen species (ROS) and transforming growth factor ?_1 (TGF-?_1) in co-cultured human ECV304 cells, and to study the intervention with tea polyphenols (TPs). METHODS: The endothelial cells were cultured alone or co-cultured with mesangial cells in high glucose media with or without TPs for 0 h, 12 h and 36 h, respectively. The activity of SOD and the content of MDA in the media of the system were detected by spectrophotometry. The expression of TGF-?_1 mRNA in the endothelial cells was measured by using semi-quantitative reverse transcription PCR (RT-PCR). RESULTS: High glucose decreased the activity of SOD, increased the content of MDA and up-regulated the expression of TGF-?_1 mRNA in co-cultured ECV304 cells and the effect became more prominent than the single-cultured cells. TPs interrupted it more effectively. CONCLUSION: These data suggest that there is interaction between mesangial cells and ECV304 cells under high glucose concentration. The interaction may markedly up-regulate the production of ROS and the expression of TGF-?_1 in co-cultured ECV304 cells. TPs may protect ECV304 cells by intervening intercellular interaction.

Objective To evaluate the effects of pregabalin combined with hydrochloric oxycodone on patients with ma-lignant neuropathic pain (MNP). Methods A total of 66 patients with MNP was divided into control group or treatment group randomly. The patients in control group received only hydrochloric oxycodone, and treatment group were treated with the combina-tion of pregabalin and hydrochloric oxycodone. Numeric rating scale (NRS) score was used to evaluate the analgesic effects. Med-ical outcomes study sleep scale (MOS-SS,Chinese version) was used to evaluate the improvement of sleep disorder. The changes of depression or anxiety were investigated by 17-item Hamilton Depression Rating Scale (HAMD-17) or Hamilton Anxiety Scale (HAMA), respectively. Side effects were accessed by Acute and Subacute Toxicity Grading Criteria of Anticancer Drugs (WHO). Results The pain control rate of treatment group was 87. 1% , which was superior to that of control group (58. 6% ) (P<0. 05). The improvement of sleep interference, and the quality and quantity of sleep in treatment group were also superior to that in control group (P<0. 05). After the treatment, depression and anxiety was attenuated in both groups, and the improvement degree in treatment group was higher than that in control group (P<0. 05). No obvious side effects were found in either groups. Conclusion The combination therapy of pregabalin and hydrochloric oxycodone is the better way to treat MNP.

Objective To build an antibody microarray based on direct labeling strategy for microalbnrninuria measurement, and evaluate it's technical potentiality for clinical application. Methods Urine samples of diabetic patients were collected. Antibody microarrays were constructed by preparation of array support, array fabrication, then protein assay and data analysis were performed. Procedure conditions for each step especially the labeling of samples were optimized. The set-ups were evaluated in terms of sensitivity, specificity and reproducibility. Urinary albumin excretion in the samples was detected by fabricated protein array, which was compared to that detected with immunoturbidimetry. Results The signal intensity was best when protein quality ratio of pure albumin or urine sample against NHS-biotin was 2:1. A calibration curve with a correlation coefficient of 0.9995 was established. The lower limit of detection was 0.0617 mg/L. Interehip and intrachip variation studies conducted on patient urine demonstrated CVs as 6.78%-9.22% and 3.35%-7.59%, respectively. Compared with the immunoturbidimetry, the antibody microarray was able to detect the extremely lower grade albumin in urine samples. The correlation coefficient of the results obtained by the two methods was 0.9199 (P <0.01). Conclusion An antibody microarray based on direct labeling strategy for microalbuminuria measurement is successfully established, which is comparable to immunoturbidimctry in its accuracy and will have great potential for clinical use with its high throughput, sensitivity, specifity and reproducibility.

Objective To survey the epidemiology of chronic kidney disease (CKD) among elderly people in two districts of Jiangsu province, China. Methods A total of 1404 residents aged 60 years or older in Huaian and Suzhou city in Jiangsu province were randomly recruited from the community population. All the people were screened for albuminuria (increased morning spot urine albumin-to-creatinine ratio, UACR) and reduced renal function (decreased eGFR by simplified MDRD equation). Urinary creatinine and albumin, serum creatinine, uric acid, cholesterol,triglyceride, low density lipoprotein-cholesterol, high-density lipoprotein cholesterol and fasting blood (1.73 m2) -1] and/or albuminuria (UACR ≥30 mg/g). The associations between healthy characteristics and indicators of kidney damage were examined. Results A total of 1316 (93.7%) elderly people completed the study. The prevalence of CKD was 32.3% and the awareness rate was only 9.6%. Albuminuria and reduced renal function were found in 30.2% and 3.2% of subjects respectively. The Logistic regression model showed that age, gender, hypertension,hypercholesterinemia and hyperuricaemia were independently associated with CKD. Systolic blood pressure (SBP) of 140-159 mm Hg exhibited a lower adjusted OR value (0.675) for CKD, while SBP of 160-179 mm Hg and of at least 180 mm Hg exhibited higher adjusted OR values (1.330 and 1.146). Similarly, FBG of 5.6-6.9 mmol/L exhibited a lower adjusted OR value for CKD as compared to FBG of at least 7.0 mmol/L (0.628 vs 1.941). Conclusions The prevalence of CKD in elderly people of Jiangsu province is quite high. Age, gender, hypertension,hypercholesterinemia and hyperuricaemia are independent risk factors for the development of CKD.

Objective To explore the factors related to the delayed graft function (DGF). Methods Clinical data of 150 recipients were collected and performed by Cox proportional hazards regression analysis . In addition, the glutathione S-transferase (GST) gene polymorphism of 172 donors and 157 healthy persons was analyzed by multiple PCR and SSP-PCR. Results DGF was observed in 24 patients among 150 recipients. Pretranplantation dialysis mode, PR A levels and recipient gender were uncorrelated with the incidence of DGF(P>0. 05). Urinary volume of the second 24 hours after transplantation was an independent predictor of DGF(RR=1. 002, P = 0. 001). The frequency of donor's null GSTM1 in DGF group was significantly higher than that in non-DGF group(P<0. 05). Conclusions Urinary volume of the second 24 hours after transplantation could be a predictor for DGF. The null GSTM1 in donor might be one of the factors related to the EGF.