Objective To silence human gene Set7/9 and screen out stable transfection cell line in hepatocellular carcinoma cell line HepG2 so as to investigate the impact of down-regulation of Set7/9 in cell line HepG2 and provide experimental foundation for studies on the effect of set7/9 in HepG2.Methods The target oligo was designed and synthesized;shRNA interference vector and the control vector were constructed and transfected into HepG2 cells;the stable transfection cells were screened out.Then Real-time PCR and Western blot were performed to detect the silence of Set7/9 according to both gene expression and protein expression level. Results The shRNA interference vector was constructed and transfected into HepG2 cells successfully.Compared with that in the negative control group,the expression of Set7/9 was dramatically downregulated (P < 0.05 ). Meanwhile,the expression of related protein Sirt1 and Suv39h1 was upregulated 8.4 folds and 1.1 fold, respectively.Conclusion Downregulation of Set7/9 expression can upregulate Sirt1 and Suv39h1,suggesting that Set7/9 may affect the activity of HepG2 cell lines.

Objective To observe the effect of traditional Chinese medicine (TCM) Qingshen granule on signal transduction pathway of fokal adhesion kinase-rat sarcoma-mitogen activated protein kinase (FAK-Ras-MAPK) in renal tissue of rats with renal interstitial fibrosis (RIF). Methods Forty Spargue-Dawley (SD) rats were randomly divided into normal control, model, Bailing capsule and Qingshen granule groups (each, n = 10). The rat model with RIF was reproduced by unilateral ureteral obstruction (UUO) or ligation method. The rats in Bailing capsule group were treated with Bailing capsule dissolved in 4 mL warm water, and the dosage was 0.3 g·kg-1·d-1 for intragastric administration;the rats in Qingshen granule group were treated with Qingshen granule dissolved in 4 mL warm water, its dosage was 6 g·kg-1·d-1 for intragastric administration, and equal volume of normal saline was given to normal control group and model group by gavage. After treatment for 8 weeks, the levels of blood urea nitrogen (BUN), serum creatinine (SCr), fibronectin (FN),α-smooth muscle actin (α-SMA) were determined in each groups. The renal tissue expression levels of FAK, Ras, p38MAPK, FN, α-SMA were determined in various groups by immunohistochemistry staining method. Results Compared with the normal control group, the levels of serum BUN, SCr, FN and α-SMA were significantly increased in the model group. There were no significant differences in the levels of serum BUN and SCr before administration of drugs between Bailing capsule group and Qingshen granule group (both P>0.05). The levels of BUN, SCr, FN, andα-SMA were all significantly lowered in the two treatment groups than those of the model group after administration, the descent in Qingshen granule group being more marked [BUN (mmol/L):13.18±4.91 vs. 18.56±5.59, SCr (μmol/L): 104.80±12.04 vs. 119.02±12.47, FN (mg/L): 29.72±16.75 vs. 46.38±8.63, α-SMA (kU/L):5.49±2.68 vs. 7.13±2.37, all P < 0.05]. The immunohistochemistry staining showed: the kidney tissue expression levels of FAK, Ras, p38MAPK, FN, and α-SMA in the model group were significantly higher than those of normal control group, above indexes were all significantly lower in Bailing capsule group and Qingshen granule group than those of the model group, and the descent in Qingshen granule group was more obvious (FAK: 3.00±1.41 vs. 5.28±2.21, FN: 4.25±1.04 vs. 6.29±2.06, α-SMA: 3.25±1.28 vs. 4.86±1.57, p38MAPK: 2.50±1.31 vs. 4.71±2.50, Ras:3.50±1.41 vs. 4.29±1.38, all P<0.05). Conclusion Qingshen granule can reduce serum BUN and SCr levels in rats with RIF, and inhibit the activation of FAK-Ras-MAPK signal transduction pathway in the kidney, thereby it may reduce the generation of FN andα-SMA and play a role of anti-RIF.

Objective To study a case series of thoracolumbar flexion-distraction fractures combined with compression of anterior and central columns to analyze the causes of its occult symptoms and explore the injury mechanism,imaging diagnosis and operative reduction of the fractures.Methods A retrospective study was carried out on data of 39 patients with thoracolumbar flexion-distraction fractures admitted into our hospital from May 2003 to December 2006.Of all,there were 17 patients with compression of anterior and central columns.The diagnostic value of imaging examinations such as X-ray,CT and MRI was evaluated.All patients were treated with posterior operation,in which simple distraction of the anterior and central columns was followed by compression and reduction of the posterior column from the rear axial direction.The clinical efficacy and safety of operation were assessed.Results The patients with compression of anterior and central columns accounted for 43.6% of all thoracolumbar flexion-distraction fractures.Before operation,six patients were misdiagnosed as simple compression fracture.Various kinds of imaging examinations detected the bone injury and/or ligaments complex injury of the posterior column at different degrees.The positive results on X-ray,CT,spiral CT multi-planar reconstruction (MPR)and MRI were in 8,7,11 and 17 patients respectively.After simple posterior distraction in 17 patients,there occurred over distraction in eight patients who were cured with posterior recompression plus reduction.Conclusions The posterior column injury of thoracolumbar flexion-distraction fractures combined with compression of anterior and central columns is relatively occult and easy to be misdiagnosed as simple compression fracture,when MPR CT and MRI are helpful for diagnosis.The operation with twostep reduction of axial distraction followed by compression can attain satisfactory and safe results.

Objective To investigate the effects of c?myc promoter binding protein 1(MBP?1)gene on the proliferation of human Saos?2 osteo?sarcoma cells in vitro. Methods Saos?2 cells were divided into three groups:blank control group(untransfected cells),negative group(cells transfected with missense sequence)and experimental group(cells transfected with MBP?1 shRNA). Two MBP?1 shRNA sequences and one neg?ative control shRNA sequence were designed ,synthesized and cloned into pSIREN?retroQ plasma. Then the recombinant plasmids were construct?ed and transfected into human Saos?2 osteosarcoma cells by Lipofectamine 2000. The expressions of MBP?1 mRNA and protein in Saos?2 cells were detected by real?time PCR and Western blot ,respectively. The effects of altered expression of MBP?1 on cell proliferation were measured by CCK?8 cell proliferation assay. The expressions of cyclin D1 and cyclin E in Saos?2 were determined by Western blot. Results PCR and sequenc?ing results indicated that the recombinant plasmids pSIREN?retroQ was constructed. The relative expression level of MBP?1 mRNA in the MBP?1 siRNA transfection group was significantly decreased than that in blank control group(P<0.05). Compared with the blank control group,the ex?pression levels of MBP?1 protein in the experimental group also significantly decreased. The proliferation abilities of Saos?2 cells at 48,72,and 96 hours after MBP?1 siRNA transfection were significantly increased than those in the blank control group(P<0.05). Compared with the blank con?trol group,the expression levels of cyclin D1 and cyclin E protein in the experimental group also significantly increased(P<0.05). Conclusion Knockdown of the expression of MBP?1 gene promotes the proliferation of human Saos?2 osteosarcoma cells. MBP?1 gene may become the new tar?get of gene therapy for osteosarcoma.

Objective To observe the improvement effects of Qingshen Granule on the life quality of chronic kidney disease (CKD) patients with damp-heat syndrome. Methods The CKD patients with damp-heat syndrome were randomly divided into experimental group and control group. The control group was treated with Western medicine, and the treatment group was treated with Qingshen Granules additionally for 12 weeks. SCr and eGFR were observed before and after treatment, and the life quality was investigated by KDQOL-SFTM1.3. Results Actually 156 cases were completed, including 77 cases in the treatment group and 79 cases in the control group. The total rate of curative effect was 81.82% (63/77) in teatment group, and (63.29%, 50/79) in control group, with significent difference (P<0.05). Multivariate linear regression analysis indicated that gender, education level, TCM syndrome score, CKD stage, eGFR and Hb level could affect the life quality. The scores of KDQOL-SFTM and its containing the MOS item short from health survey (SF-36) and kidney disease targeted areas (KDTA) were significantly improved after treatment in experimental group (P<0.05), while the scores in control group were with no significant improvement. The improving effects of PCS, and SPL, SLEEP, PS in KDTA in treatment group were significantly superior to those in control group (P<0.05). The scores of life quality in effective and stable cases increased significantly after treatment compared with before treatment (P<0.05). Conclusion Qingshen Granules can effectively improve the life quality of CKD patients with damp-heat syndrome, and the effect is not dependent on the improvement of laboratory indexes (renal function).

OBJECTIVE: To evaluate the therapeutic effect of Qingshen Granules on renal anemia in patients with damp-heat syndrome and explore the mechanisms in light of inflammation/hepcidin axis and iron metabolism. METHODS: Sixty patients with renal anemia and dampness-heat syndrome were randomized into control group (=30) and treatment group (=30). All the patients were given routine treatment, and the patients in the treatment group received additional treatment with Qingshen Granules (3 times a day). After 12 weeks of treatments, the patients were examined for changes in the integral value of TCM syndrome, serum creatinine (Scr), glomerular filtration rate (eGFR), hemoglobin (HGB), hematocrit (HCT), red blood cell (RBC) count, interleukin-6 (IL-6), hypersensitive C-reactive protein (hs-CRP), ferritin, growth differentiation factor-15 (GDF-15), serum iron (SI), total iron binding capacity (TIBC), transferrin saturation (TAST), soluble transferrin receptor (sTfR) and ferritin levels. RESULTS: After the treatment, the scores of TCM syndrome were significantly improved in the treatment group and were better than those in the control group (=0.000). Scr and eGFR were improved in both groups after the treatment. The levels of HGB, HCT and RBC were all improved in the two groups after treatment, and the improvements were more obvious in the treatment group (=0.002, 0.002, and 0.017, respectively). The levels of IL-6, hs-CRP, hepcidine and GDF-15 were all lowered in the two groups after the treatment, and they were all significantly lower in the treatment group than in the control group (all =0.000). The treatments increased the levels of SI and TAST in both of the groups, and compared with those in control group, the levels of TIBC, sTfR and ferritin were significantly lowered in the treatment group after the 12-week treatment (=0.000). CONCLUSIONS Qingshen granules can effectively improve renal anemia in patients with damp-heat syndrome possibly by improving iron metabolism through alleviation of inflammation and reduction of hepcidine level.

OBJECTIVE: To observe the effect of Ronghuang granule on serum fibroblast growth factor 23 (FGF23), fibroblast growth factor receptor (FGFRs) and Klotho protein levels in non-dialysis patients with chronic kidney disease-mineral and bone disorder (CKD-MBD) and kidney deficiency and damp heat syndrome. METHODS: Seventy non-dialysis CKD-MBD patients with kidney deficiency and dampness-heat syndrome were randomized into control group (=35) and treatment group (=35). All the patients were given routine treatment combined with traditional Chinese medicine retention enema, and the patients in the treatment group received additional Ronghuang granule treatment (3 times a day). After the 12-week treatments, the patients were examined for changes of TCM syndromes. Serum levels of Ca, P, parathyroid hormone (iPTH), FGF23, FGFRs and Klotho proteins were detected before and after treatment. These parameters were also examined in 20 healthy volunteers. RESULTS: Sixty-five patients completed the study, including 33 in the control group and 32 in the treatment group. The patients in the treatment group showed significantly better treatment responses than those in the control group ( < 0.05 or 0.01). At 4, 8, and 12 weeks of treatment, the patients in the treatment group had significantly lowered scores of TCM syndromes compared with the score before treatment ( < 0.05 or 0.01), while in the control group, significant reduction of the scores occurred only at 12 weeks ( < 0.05); at each of the time points, the treatment group had significantly greater reductions in the score than the control group ( < 0.01). Significant improvements in serum Ca, P and iPTH levels were observed at 4, 8, and 12 weeks in the treatment group ( < 0.05) but only at 12 weeks in the control group ( < 0.05). The patients in the control and treatment groups all showed elevated serum levels of FGF23, FGFRs and Klotho protein compared with the normal subjects ( < 0.01); FGF23, FGFRs and Klotho levels were significantly reduced in the treatment group ( < 0.05) but remained unchanged in the control group (>0.05), showing significant differences between the two groups. CONCLUSIONS Ronghuang granule improves the clinical symptoms of non-dialysis CKD-MBD patients with kidney deficiency and dampness heat syndrome by reducing serum levels of FGF23, FGFRs and Klotho, improving calcium and phosphorus metabolism disorder, and inhibiting secondary hyperparathyroidism.
Calcium ; blood ; Chronic Kidney Disease-Mineral and Bone Disorder ; blood ; therapy ; Drugs, Chinese Herbal ; pharmacology ; Enema ; Fibroblast Growth Factors ; blood ; Glucuronidase ; blood ; Humans ; Parathyroid Hormone ; blood ; Phosphorus ; blood ; Receptors, Fibroblast Growth Factor ; blood ; Renal Insufficiency, Chronic ; blood ; therapy ; Sweating Sickness ; blood ; therapy ; Syndrome