AIM: To construct recombinant adenoviral vector carrying the LEDGFp52 gene by homologous recombination in bacteria and to detect its expression in vitro.METHODS: The LEDGFp52 gene was cloned to adenoviral shuttle plasmid pAdTrack-CMV. Then, the resultant pAdTrack-CMV-LEDGFp52 was cotransfected into BJ5 183 bacteria with the adenoviral backbone plasmid pAdeasy-1. The adenoviral plasmid carrying LEDGFp52 was generated with homologous recombination in bacteria, and the adenoviruses were produced in 293 cells. These 293 cells were then infected with adenoviruses, and the expression of LEDGFp52 was detected by CPE (cytopathic effect) and western blot.RESULTS: The titer of Ad-LEDGFp52 adenoviruses was up to 5×1012 pfu/L after proliferation in 293 cells. LEDGFp52 was expressed efficiently in 293 cells after infection.CONCLUSION: The recombinant adenoviruses vector expressing LEDGFp52 was constructed successfully and can be used in further gene transfection experiments.

AIM: To construct and identify LEDGFp52 eukaryotic expression vector for RNA interference.METHODS: Recombinants were designed and established by targeting gene LEDGFp52 and plasmid pGensil-1 based on LEDGFp52 cDNA sequences of Genomes. Two pairs of oligonucleotides were synthesized according to the Tuschl principle and inserted into plasmid pGenSil-I to generate siRNA eukaryotic expression vector. DH5α strains were transformed, plasmids were extracted, and recombinant vectors were identified by the restriction map and the sequence analysis. The cultured cells were transfected by the recombinant plasmid (pGensil-1-RNA. LEDGFp52-1). At 48 hours after transfection, the whole cell protein was extracted, and the protein level was detected using Western blotting with mouse anti-human LEDGFp52 monoclonal antibody.RESULTS: Recombinant plasmids completely concord with the designs by the restriction map and the sequence analysis,the proteinlevel of LEDGFp52 was down regulated at 48hours after transfecting pGensil-1- LEDGFp52-1 expression vector into HeLa cells, the recombinant eukaryotic expression vectors were successfully constructed.CONCLUSION: siRNA recombinant can be successfully constructed by RNAi technique to inhibit the expression of LEDGFp52.

AIM: To construct the prokaryotic expression vector of rhLEDGFp52 gene,to obtain the rhLEDGF p52 protein.METHODS: rhLEDGFp52 gene was constructed into a prokaryotic expression vector pET30a (+) by recombinant DNA techniques and was identified by enzymatic digestion and sequence analysis. rhLEDGFp52 protein was induced expression by IPTG in E.coli BL21 (DE3), it was tested by Western blot and was purified by Ni-NTA His. Bind. Resin.RESULTS: We Successfully constructed the prokaryotic expression vector of rhLEDGFp52 gene and obtained its expression in E.coli BL21 (DE3),it was expressed in a soluble form and detected up to 34.63% of the total bacterial protein expressed in E.coli BL21 (DE3).Western blot analysis demonstrated that rhLEDGFp52 protein could spicifically integrate with LEDGF-ab. After purified by Ni-NTA His. Bind. Resin, the ultimate concentration of purified rhLEDGFp52 protein was 520μ g/ml and its purity was 87.93%.CONCLUSIONS: rhLEDGF p52 protein was obtained that provides an experimental basis for the further study of the biological function of rhLEDGFp52 protein.

Objective To investigate the changs of serum and urine vascular endothelial growth factor(VEGF) in children with steroid-sensitive nephrotic syndrome(SSNS) at their active and remission stage.Methods Serum and morning urine levels of VEGF were assayed by Liquichip in 30 patients with SSNS at the active and remission stage and 30 normal age-and sex-matched controls.Results Serum levels of VEGF in patients at the active stage [(186.62?106.21) ng/L] were significantly higher those that at the remission stage [(118.75?73.08) ng/L] and than those in control group [(108.64?54.75) ng/L](P0.05).Morning urine levels of VEGF in patients at the active stage [(201.66?100.46) ng/L] were significantly higher than those at the remission stage [(116.35?55.99) ng/L] and than those in control group [(99.94?42.07) ng/L](P0.05).Conclusions Serum and morning urine levels of VEGF are significantly elevated in patients with SSNS,however they are significantly lower after the treatment of steroid.There is no obvious difference between the patients at the remission stage and healthy controls.VEGF involves pathogenetically and physiologically in SSNS during its onset and growth.

Objective To retrospectively analyze the clinical data of 16 patients with renal cell carcinoma complicatedwith sarcomatoid differentiation, so as to discuss the treatment and prognosis of the condition. Methods A total of 547 patients with renal cell carcinoma were admitted in our department during 2005-2009, and 16(2. 9%) ofthem also had sarcomatoid differentiation, including 7 males and 9 females, aged (52 ± 10) years old, ranging 36-70 years old. Gross hematuria was found in 5 patients, backache in 5, fatigue and weight loss in 2, cough and hemoptysis in 1; and 3 patients were accidentally found with renal occupation during physical examination. All tumors were solitary, with 7 in the left and 9 in right. The maximum tumor diameter was (7. 8±4. 1) cm, ranging 2-15 cm. Full examination was done to confirm the tumor stage preoperatively. Preoperative examination showed that 2 patients had retroperitoneal lymph node enlargement, 1 had right pulmonary hila lymphatic metastasis, 1 had livermetastasis, 2 had renal vein and (or) the inferior vena cava tumor thrombi, 3 had bone metastases, 1 had invasion of ipsilateral adrenal gland, and 1 had invasion of the upper ureter. All the 16 patients underwent surgical treatment,and their specimens were subjected to H-E staining and immunohistochemical examination. Five patients were treated with INF-α after operation and were followed up. Results All patients received radical nephrectomy. Pathological examination showedthat 9 patients had clear cell renal cell carcinoma with sarcomatoid differentiation, 3 had chromophobe renal cell carcinoma with sarcomatoid differentiation, and the other 4 had unclassified renal cell carcinoma with sarcomatoid differentiation. No recurrence or metastasis was seen in the 4 pT1N0M0 patients. All the pT2-T4 patients died, with the progression-free survival time being (5. 6±4. 5) months, ranging 2-16 months; the overall survival time was (8. 3± 5. 6) months, ranging 3-20 months. Conclusion Preoperation staging of patients with renal cell carcinoma complicated with sarcomatoid differentiation has a great impact on patient survival. Operation is the most effective treatment for patients with early stage, and routine follow? up examination is recommended post operatively. Targeted therapy may be an effective way for patients of advanced stag.

To retrospectively analyze the clinical data of 16 patients with renal cell carcinoma complicated with sarcomatoid differentiation, so as to discuss the treatment and prognosis of the condition. Methods A total of 547 patients with renal cell carcinoma were admitted in our department during 2005-2009, and 16(2.9%) of them also had sarcomatoid differentiation, including 7 males and 9 females, aged (52±10) years old, ranging 36-70 years old. Gross hematuria was found in 5 patients, backache in 5, fatigue and weight loss in 2, cough and hemoptysis in 1; and 3 patients were accidentally found with renal occupation during physical examination. All tumors were solitary, with 7 in the left and 9 in right. The maximum tumor diameter was (7.8±4.1) cm, ranging 2-15 cm. Full examination was done to confirm the tumor stage preoperatively. Preoperative examination showed that 2 patients had retroperitoneal lymph node enlargement, 1 had right pulmonary hila lymphatic metastasis, 1 had livermetastasis, 2 had renal vein and (or) the inferior vena cava tumor thrombi, 3 had bone metastases, 1 had invasion of ipsilateral adrenal gland, and 1 had invasion of the upper ureter. All the 16 patients underwent surgical treatment, and their specimens were subjected to H-E staining and immunohistochemical examination. Five patients were treated with INF-α after operation and were followed up. Results All patients received radical nephrectomy. Pathological examination showed that 9 patients had clear cell renal cell carcinoma with sarcomatoid differentiation, 3 had chromophobe renal cell carcinoma with sarcomatoid differentiation, and the other 4 had unclassified renal cell carcinoma with sarcomatoid differentiation. No recurrence or metastasis was seen in the 4 pT1N0M0 patients. All the pT2-T4 patients died, with the progression-free survival time being (5.6±4.5) months, ranging 2-16 months; the overall survival time was (8.3± 5.6) months, ranging 3-20 months. Conclusion Preoperation staging of patients with renal cell carcinoma complicated with sarcomatoid differentiation has a great impact on patient survival. Operation is the most effective treatment for patients with early stage, and routine follow-up examination is recommended postoperatively. Targeted therapy may be an effective way for patients of advanced stage.

Abnormal activation of Wnt signaling pathway plays an important role in the development and progression of prostate cancer; androgen receptor (AR) is the key for the transformation of androgen-independent prostate cancer (AIPC), and AR signaling pathway is now the focus of prostate cancer research. Many signal pathways can affect AR signaling pathway; in this paper we review the Wnt signaling pathway, androgen receptor and their crosstalk in prostate cancer.

Alcohol Drinking ; adverse effects ; Cardiovascular Diseases ; etiology ; prevention & control ; Health Promotion ; Humans ; Risk Assessment

Objective To study whether clinical variables could be used to predict the presence of esophageal varices(EV). Methods One hundred and twenty-six patients with decompensated liver cirrhosis were enrolled. Upper endoscopy was performed to identify the EV. The spleen vein (SV), portal vein(PV), spleen index(SI), ascites was determined by ultrasenography. Platelct count(Pt), prothrombin time(PT) and liver function was determined. Results Ninety-five patients with EV, and 42 patients with severe EV. Patients with EV had significant larger SI and lower Pt. Pt and SI were predictive factors for the presence of EV. When SI≥66.9 cm2 and Pt≤89.0×109/L, they had a positive predictive value of 97.4% and 96.5%, and a negative predictive value of 55.4% and 59.8%, respectively. SI was the only predictive factor for the presence of severe EV. When SI≥82.6 cm2, it had a positive and negative predictive value of 89.2% and 75.4%. Conclusions Pt and SI are predictive factors for the presence of EV. SI is the only predictive factor for the presence of severe EV. Non-invasive factors SI and Pt can be used to predict the EV in patients with decompensated liver cirrhosis.

Age Factors ; Aged ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Female ; Humans ; Male ; Middle Aged