Objective To observe the anti-tumor effect and mechanism of curcumin on triple-negative breast cancer mouse model.Methods Human breast cancer cells MDA-MB-231 were inoculated to the chest subcutaneous fat pad of mice(about 1 million cells in 0.1 mL cell suspension).At day 2 after tumor inoculation,nodules were seen at inoculation site (tumor formation rate being 100％).Forty successfully-modeled mice were randomly divided into low-,middle-,and high-dose curcumin groups (10,20,40 mg/kg) and model control group,10 mice in each group.The medication lasted for 30 continuous days.After medication,the blood was taken out from the orbital venous plexus,and the serum was separated for the detection of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) contents.Spleen was separated for the calculation of organ coefficient,tumor mass was weighed,and the expression levels of Bcl-2 and caspase-3 were detected by Western blotting method.Results Compared with the model control group,caspase-3 expression level was significantly increased in the three curcumin groups(P ＜ 0.05 or P ＜ 0.01),but the contents of TNF-α and IL-6,tumor weight,spleen index and Bcl-2 expression were significantly decreased(P ＜ 0.05 or P ＜ 0.01).Conclusion Curcumin has inhibitory effect on triple-negative breast cancer mouse model through promoting the apoptosis of breast cancer cells.

Objective To study the prevalence and genotypes of rotavirus (RV) among children,< 5 years old hospitalized with viral diarrhea in Tianjin. Methods Stool specimens were collected from hospitalized diarrhea children in Tianjin children's hospital between May 2008 and April 2009. Detection of rotavirus was employed by Colloidal Gold Device. The detected positives were inoculated to MA-104 cells. The total RNA of virus was extracted after CPE which was caused by rotavirus were observed, The VP7 serotypes were determined by using RT-PCR to amplify the VP7 gene and sequencing the RT-PCR products.The clinical data for each patient were also collected. Results Among 837 specimens, the RV antigen positive rate was 26. 3% (220/837). Among all the children with rotavirus diarrhea, 90. 5% (199/220)were < 2 years old. The prevalence of rotavirus diarrhea in children peaked during Oct. 2008 through Apr.2009. Of the 208 rotavirus positive specimens, 95 were successfully identified by RT-PCR Thirty-five positive strains of RV were sequenced, and the sequencing results showed that 32 positive strains were belonged to rotavirus G1 type, 2 positive strains were belonged to rotavirus G3 type and 1 positive strain were belonged to rotavirus C9 type. Conclusion RV was the dominant etiological agent for infantile diarrhea infection in Tianjin, and the predominant serotype was G1.

Objective To study the pathogenic prevalence and genotypes of astrovirus among children under 5 years old hospitalized with diarrhea in Tianjin. Methods A total 837 stool specimens were collected from children with diarrhea hospitalized in Tianjin children's hospital from May 2008 to April 2009. Astrovirus antigens were detected using ELISA and the postive specimens were inoculated in CaCo-2cells. After the CPE caused by virus were observed, the total RNA of virus was extracted, then the genomc fragments of the strains were amplified by using RT-PCR and confirmed by sequencing of the RT-PCR products. Detection of rotavirus was employed by Colloidal Gold Device. Results Astrovirus antigen was found positive in 3.0% of the patients. The coinfection rate of astrovirus and rotavirus was 0. 7% (6/837).Ninety-six persent of children with astrovirus diarrhea were younger than 2 years of age, Forty-eight persent of children with astrovirus diarrhea were younger than 6 months. The astrovirus infections occurred mainly between August 2008 and April 2009. Of the 21 astrovirus positive specimens, 11 cases were successfully identified by RT-PCR and they were all serotype 1. Conclusion Astrovirus is a major cause of nonbacterical diarrhea between 2008 and 2009 in Tianjin, and the predominant serotype is type 1.

Objective To analyze the epidemiographic features of urological de novo malignant tumor in kidney transplant recipients in the General Hospital of Jinan Military Command.Methods The clinical data of 1945 patients who received kidney transplantation between September 1978 and December 2009 were retrospectively studied.Among 1945 recipients,22 cases were diagnosed as having urological de novo malignant tumors ( incidence:1.13％ ),including renal papillary adencaicinoma (n =1 ),papillary renal cell carcinoma (n =1 ),renal hemangiosarcoma (n =1 ) ; pelvic transitional cell carcinoma (TCC) (n =1 ),pelvic and ureter TCC (n =6),ureter TCC (n =7),pelvic and ureter and urinal bladder TCC (n =1 ),4 cases of bladder malignant tumors (including 3 cases of bladder TCC and 1 case of borderline bladder tumor).Of the 22 cases,17 had a main clinical manifestation of gross hematuria and 2 had microscopic hematuria,and the rest 3 had no obvious symptom.The average age at diagnosis of these 22 cases was 54.3 ± 12.3 years,with a mean time of 53 months after kidney transplantation.Ten cases received immunosuppressive treatment by using cyclosporine A (CsA) + azathioprine (Aza) + prednisone (Pred),while the remaining 12 received CsA + MMF + Pred.Surgical treatment was carried out in all cases:radical nephrectomy was conducted for 3 cases of renal carcinoma; total resection of kidney,ureter and sleeve-shaped resection of bladder in affected side were conducted for the 15 cases of pelvis or ureter carcinoma; for the 4 cases of bladder carcinoma,transurethral resection of bladder tumor was conducted for 3 cases while partial cystectomy was conducted for the other one case.Results During a follow-up period of 2 to 97 months,there were 9 deaths 6 to 97 months after toumorectomy.One died of bone metastasis,one pulmonary metastasis,two brain metastasis,two hepatic metastasis,and three extensive metastatic tumor soon after the diagnosis.Thirteen patients survived through the follow-up,with the longest survival time being 92 months in one patient with urinary bladder tumor.Four patients survived longer than 4 years,and 5 cases longer than 1 year.Conclusion Urological de novo malignant tumor is an important complication after renal transplantation with a main clinical manifestation of painless gross hematuria,and surgical resection is the most important treatment.

Objective To summarize the clinical characteristics,diagnosis and treatment of malignant tumor after renal transplantation.Methods The clinical data of 2106 renal transplants in 1945 patients undertaken in our hospital from September 1978 to December 2009 was retrospectively studied.Results Of these 1945 patients, 39 cases were diagnosed as having malignant tumor (incidence: 2.0 ％).The interval between transplantation and clinical diagnosis ranged from 8 to 124 months with a median of 57.0 months.Among the 39 cases of malignancy, there were 22 urinary system carcinomas, 8 digestive system carcinomas,2 lung cancers,2 breast cancers,2 lymphomas,1 dura small cell carcinoma,1 pleura poorly differentiated carcinoma and 1 metastatic carcinoma of liver with unknown primary tumor. Surgery was conducted in 28 patients,of which 16 were survived but the other 12 patients died of metastasis ranged from 3 months to 96 months (median,33 months) after operation.11 cases without operation died within from 3 d to 36 months (median,5 months) after diagnosis.Conclusions The incidence of malignant tumors in renal transplant recipients increased markedly.The most common type of the malignant tumors is urinary system carcinoma.The key measure of success in treating malignancy after renal transplantation is early diagnosis and surgical resection.

AIM: To investigate the effect of silymarin on homocysteine-induced cell viability and apoptosis in human umbilical vein endothelial cells (HUVECs). METHODS: Cell viability was analyzed by using MTT and LDH assay. Apoptotic cells were detected by using DNA fragmentation and flow cytometric analysis. The level of intracellular reactive oxygen species (ROS) and the potential of mitochondrial membrane were determined by flow cytometric assay. The activity of caspase-3, -6 and -9 were measured with microplate spectrofluorometer. Protein levels were examined by Western blotting. RESULTS: Treatment of cultured HUVECs with HCY for 48 h induced a significant decrease in cell viability, and the percentage of apoptosis increased to 76.8%. The level of intracellular ROS and activity of caspase-3, -6 and -9 enhanced, and the red/green ratios of mitochondrial membrane decreased. However, simultaneous treatment with silymarin exhibited cytoprotective effects, reduced formation of the DNA ladder, prevented the levels of Bax and Bcl-2 proteins and the accumulation of ROS as well as caspase-3, -6 and -9 activation, reconverted the potential of mitochondrial membrane, and the percentage of apoptosis/necrosis was significantly decreased to 12.7% in a dose-dependent manner. CONCLUSION: These results demonstrate that silymarin has the protective capacity to antagonize HCY-induced apoptosis in HUVECs. The antiapoptotic action of silymarin may be partially dependent on an anti-oxidative stress effects, inhibition of caspases activity, and maintenance of mitochondria function.

BACKGROUND:Thrombospondin-1 is an important endogenous activator of transforming growth factor beta 1 in this experimental inflammatory kidney disease model. Transforming growth factor beta 1 is considered the major cytokine that causes tissue fibrosis in many different inflammatory disease processes, in particular in renal disease.
OBJECTIVE:To investigate the expression of thrombospondin-1 on renal fibrosis in rats.
METHODS:Healthy male Sprague-Dawley rats were randomly divided into sham surgery group and model group. In themodel group, right ureters of rats were ligated to construct models of renal fibrosis. 3 weeks after surgery, blood and urine were obtained weekly. Enzyme linked immunosorbent assay and Bradford method were used to detect the contents of serum creatinine,blood urea nitrogen and urinary protein. After rats were sacrificed, kidneys were fixed. Western blot assay was utilized to identify the expression of vascular endothelial growth factor, transforming growth factor beta 1 and thrombospondin-1 protein. Hematoxylin-eosin staining was applied to detect the changes in pathological structure of the kidney after surgery.
RESULTS AND CONCLUSION:(1) One week after model induction, urinary protein, serum creatinine and urea nitrogen levels were significantly higher in the model group than in the sham surgery group (P< 0.05). Three weeks later, the difference in each index was significant (P< 0.01), which showed that the injury of the kidney was aggravated. (2) Transforming growth factor beta 1 protein and thrombospondin-1 expression was significantly higher in the model group than in the sham surgery group, but vascular endothelial growth factor protein expression was significantly lower in the model group than in the sham surgery group. (3) Hematoxylin-eosin staining results demonstrated that severe pathological changes of renal tissue in rats were detected after ligation of renal tubule. (4) These results confirmed that thrombospondin-1 expression increased in renal tissue, and its expression was strongly associated with vascular endothelial growth factor protein and transforming growth factor beta 1, which may play an important role in the renal fibrosis.

BACKGROUND:Type 1 diabetes mel itus is an autoimmune disease, which is characterized as the selective destruction of pancreatic beta cel s in the body, resulting in the lack of insulin secretion. Umbilical cord blood stem cel s have the potential to differentiate into islet cel s in vitro and in vivo, which play a certain hypoglycemic effect. OBJECTIVE:To explore the effects of umbilical cord blood stem cel s on blood glucose levels and PDX-1 and MafA levels in the pancreatic tissue of type 1 diabetic rats. METHODS:Thirty Sprague-Dawley rats were randomly divided into three groups, with 10 rats in each group. In treatment and model groups, type 1 diabetes mel itus modes were established. After modeling, the treatment group was given a single tail vein injection of umbilical cord blood stem cel s;the normal control group was given the same volume of normal saline;the model group was given the same volume of umbilical cord blood stem cel buffer solution. Oral glucose tolerance test was adopted to assess the islet function of rats;hematoxylin-eosin staining was used to observe the pancreatic morphology of rats;western blot and PCR methods were employed to detect expressions of PDX-1 and MafA in pancreatic tissues at protein and mRNA levels. RESULTS AND CONCLUSION:(1) Compared with the normal control group, the levels of blood glucose in the model and treatment groups were significantly higher at 0, 30, 60, 90 minutes (P<0.05). At 120 minutes, the blood glucose level in the model group was significantly higher than that in the normal control group (P<0.05), but there was no difference between the treatment and normal control groups (P>0.05). (2) The number of islets in the model group was decreased, and the boundary was unclear and irregular;in the treatment group, the number of islets was decreased, but the boundary was stil clear. (3) The expressions of PDX-1 and MafA in the treatment group were similar to those in the normal control group (P>0.05), but significantly higher than those in the model group (P<0.05). These findings suggest that the umbilical cord blood stem cel transplantation can significantly reduce the blood glucose levels in type 1 diabetic rats, improve the function of islet and morphology of pancreas, and up-reuglate the expressions of PDX-1 and MafA.

BACKGROUND:Common strategies for preventing diabetic nephropathy include effective control of blood sugar and blood pressure, inhibition of the rennin-angiotensin system and lipid-lowering therapy, but it is often difficult to get the desired results. OBJECTIVE:To investigate the effect of transplantation of bone marrow mesenchymal stem cels on levels of blood glucose and urinary total protein in diabetic nephropathy rats. METHODS: Forty-five Sprague-Dawley rats were randomly divided into three groups (n=15 per group): normal control group, diabetic nephropathy group and stem cel transplantation group. Rats in the diabetic nephropathy and stem cel transplantation groups were given single use of 60 mg/kg streptozotocin to make diabetic nephropathy models. The same dose of citric acid-sodium citrate buffer was injected in the normal control group. After modeling, 200μL of bone marrow mesenchymal stem cel solution (2×106) was injected into the left ventricle of rats in the stem cel transplantation group, and then at 7 days after the first transplantation, the cel transplantation was conducted again. The same dose of serum-free L-DMEM was injected intracardialy into the rats in the normal control and diabetic nephropathy groups. Levels of urinary total protein and blood glucose were detected. RESULTS AND CONCLUSION:At 1, 4, 8 weeks after treatment, the urinary total protein and blood glucose levels were significantly higher in the stem cel transplantation group and diabetic nephropathy group than the normal control group (P < 0.05). At 1 week after treatment, the urinary total protein and blood glucose levels were significantly lower in the stem cel transplantation group than the diabetic nephropathy group (P < 0.05). At 4 and 8 weeks after treatment, the total urinary protein and blood glucose levels were slightly higher in the diabetic nephropathy group than the stem cel transplantation group, but there was no significant difference (P > 0.05). These findings indicate that bone marrow mesenchymal stem cel transplantation in diabetic nephropathy rats can get good results in a short period, significantly improve the blood glucose and urinary total protein levels, but the long-term treatment effect is poor.

Objective To get the gene encoding extracellular domains of gB protein of B virus and analyze its expression in the eukaryocyte cell.Methods synthesizing gene fragment encoding extracellular domains of gB protein of B virus was by using synthesis gene, then digested with the restriction endonucleases BamHⅠand NotⅠand inserted into eukaryotic expressing vector pEGFP-N3.pEGFP-N3-GB合 was transfected into 293 cells.After protein extraction, the expression of gene was detcted by western blotting, and the cellular localization of the gene was analyzed by immunofluorescence and laser scanning confocal microscopy.Results pEGFP-N3-GB合were expressed in 293 cells and on the cell membrane.Conclusion eukaryotic expressing system can produce specific antigen recombination protein of B virus gB protein and express on the cell membrane.