Background and purpose:MicroRNA (miRNA, miR) plays an important regulatory role in cancer. miR-222 is reported to be up-regulated in various tumors, but its role in renal cell carcinoma (RCC) remains unclear. In this study, we detected the expression of miR-222 in both RCC and adjacent tissue samples. The aim of this study was to investigate the role of miR-222 in RCC. Methods:The expression levels of miR-222 in RCC tissue samples were quantified by quantitative real-time polymerase chain reaction (qRT-PCR). DDIT4 and LC3-Ⅱ protein expressions were determined by Western blot. Dual luciferase assay was performed to verify the target of miR-222. EGFP-LC3 microscopy assay was performed to assess autophagy. Results:Results from qRT-PCR showed that the expression of miR-222 was up-regulated in RCC tissues. Knockdown of miR-222 with speciifc antagomiR decreased the cell viability of 786-O cells, whereas overexpression of miR-222 increased the cell viability (P<0.01). The levels of DDIT4 were up-regulated in 786-O cells transfected with miR-222 antagomiR, whereas overexpression of miR-222 induced the down-regulation of DDIT4 expression. Data from dual luciferase assay indicated that miR-222 directly targeted the expression of DDIT4. Consistently, the expression of DDIT4 in RCC tissues was down-regulated compared with adjacent tissues. Knockdown of miR-222 in 786-O cells induced a signiifcant increase of autophagosome formation and LC3 lipidation.These results supported that miR-222 could inhibit autophagy in RCC cells, which may affect the clinical characteristcs of RCC. Conclusion: miR-222 is up-regulated in RCC and can inhibit the autophagy of RCC cells through down-regulating the expression of DDIT4.

AIM:To observe the effect of Wnt/β-catenin signaling pathway on diabetic ulcer.METHODS:Diabetic animal model was established in the female Wistar rats by intraperitoneal injection of low-dose streptozotocin fol-lowing high-fat diet feeding.A circular wound was made on the dorsum of the rats in both control group and diabetic group. The condition of wound healing was recorded and the structures of the wound tissues were observed by HE staining in the 2 groups at 3, 7 and 14 d after wounding.The expression ofβ-catenin, GSK-3βand Rspo-3 at mRNA and protein levels in the wound tissues was detected by RT-PCR and ELISA.RESULTS:In diabetic group, the wound healing rate was lower (P<0.05), and the inflammatory cells, fibroblast cells and new capillaries in the wound tissues were fewer than those in control group.The expression of β-catenin and Rspo-3 at mRNA and protein levels in the wound tissues in control group was significantly higher than those in diabetic group, and the expression of GSK-3βwas exactly the opposite (P<0.05). CONCLUSION:The down-regulation of Wnt/β-catenin probably resultes from the decreased level of Rspo-3, which may be one of the reasons for delaying the diabetic ulcer healing.

ObjectiveTo explore the relationship between chronic kidney disease (CKD) and cerebral small vessel disease (SVD) in elderly patients. MethodsOne hundred and fifty-two elderly male CKD patients for experimental group and 158 elderly male for control group were recruited. Demographic data and vascular risk factors were recorded. White matter lesion (WML) was semi-quantitatively assessed by cerebral magnetic resonance imaging (MRI), and lacunar infarction (LI) was also calculated. Results(1) The prevalenees of hypertentsion and diabetes mellitus were higher in elderly CKD patients than those in control group (30. 9% vs. 19.0%, 23.7%vs. 14.6%;both P～0. 05). (2) The percentages of grade 2 and grade 3 WMLs were higher in elderly CKD patients than those in control group (34.9% vs. 24.1%, 25.7% vs. 16.5%;both P<0.05). Prevalence of LI was higher in elderly CKD patients than that in control group (45.4% vs.25.3% ,X2= 13. 70, P<0. 05). The similar Resultswere also obtained except for control subjects with hypertension and diabetes mellitus. (3) The logistic regression analysis showed that age, hypertension and low glomerular filtration rate (GFR) were closely associated with SVD in elderly CKD patients. ConclusionsHypertention and diabetes mellitus are risk factors for CKD in elderly patients. SVD is associated with CKD, and age, hypertension and low GFR may be risk factors for SVD in elderly CKD patients.

Objective: To investigate the effects of hinokitiol on the proliferative inhibition and apoptosis induction in human clear cell renal cancer 786-O cells. Methods:CCK-8 assays were performed to analyze the effects of hinokitiol on the proliferation of 786-O cells. The apoptosis rate was determined by flow cytometry. EGFP-LC3 microscopy assays were performed to assess the autoph-agy flux. Cleaved Caspase-3, LC3, and P62 were detected by Western blot. Results: Hinokitiol could inhibit the proliferation of the 786-O cells and could induce cell apoptosis via Caspase pathway. Hinokitiol induced the autophagy of 786-O cells, increased LC3 ex-pression, and downregulated P62 expression. Conclusion: Hinokitiol can inhibit the proliferation of 786-O cells and can induce cell apoptosis via autophagy induction.

BACKGROUND:Cuscuta chinensis is a mature seed of Cuscutachinensis Lam., can warm kidney. Previous studies demonstrated that kidney compound composed of Cuscuta chinensis could apparently inhibit bone loss and improve bone density.
OBJECTIVE:To explore the influence of flavonoids from Cuscuta chinensis on bone mineral density of femur, 1,25(OH)2D3 levels of serum and kidney, the expression of smal intestine CaBp-D9K mRNA in model rats with ovariectomized osteoporosis.
METHODS:A total of 72 Sprague-Dawley female rats were equal y and randomly divided into six groups (n=12):sham surgery group, model group, vitamin D3 group and low-, moderate-and high-dose flavonoids groups. The sham surgery group only received sham operation and the other five groups were ovariectomized respectively. One week after ovariectomy, the rats were given flavonoids from low-, moderate-and high-dose Cuscuta chinensis and vitamin D3 (2 mg/kg) by intragastric administration for 3 consecutive months. Blood was obtained from the abdominal aorta. Serum was isolated. The kidney was obtained. Enzyme linked immunosorbent assay was used to determine 1,25(OH)2D3 contents of renal and serum. Rats were sacrificed at the end of experiment. The thighbone was taken out to determine bone mineral density. The second lumbar vertebra was taken out to measure the expression of lumbar vertebra and renal vitamin D receptor mRNA using real-time fluorescent reverse transcription-polymerase chain reaction. The smal intestine was taken out to measure the expression of smal intestine CaBp-D9K mRNA using real-time fluorescent reverse transcription-polymerase chain reaction.
RESULTS AND CONCLUSION:Compared with the sham operation group, bone mineral density of femur, and 1,25(OH)2D3 levels of serum and kidney and the expression of lumbar vertebra vitamin D receptor mRNA significantly decreased in model group, and the expression of smal intestine CaBp-D9K mRNA significantly decreased in model group. Compared with the model group, bone mineral density of femur, and 1,25(OH)2D3 levels of serum and kidney, and the expression of the second lumbar vertebra vitamin D receptor mRNA, and the expression of smal intestine CaBp-D9K mRNA were increased in moderate-and high-dose flavonoids groups and vitamin D3 group. Results indicated that flavonoids from Cuscuta chinensis could significantly raise bone mineral density of femur, and 1,25(OH)2D3 levels of serum and kidney and the expression of lumbar vertebra vitamin D receptor mRNA and the expression of smal intestine CaBp-D9K mRNA, accelerate intestinal calcium absorption and osteoblast activity, and reinforce quality of the bone.

Objective To study the anatomy basis and biomechanical stability of euthyphoria reduction and percutaneous cannulated screw fixation for sacroiliac dislocation,and to evaluate the primary clinical efficacy of this method.Methods The distances from the anterior branches of the nerve roots at L4,and L5 and obturator nerve on the superior border of sacral ala to the sacroiliac joint were measured on 12 adult cadavers (24 sides) fixed and preserved by formalin.Models of sacroiliac dislocation were made on six pelvic specimens of fresh cadavers.A comparison of stability was made on the six models between the fixation studied here and the traditional fixations by posterior percutaneous sacroiliac screws and by anterior sacroiliac joint plates.At the same time,17 patients with type C Tile fracture were treated with our method.The clinical efficacy was analyzed for the 17 patients.Results The distances from the anterior branches of the nerve roots at L4,and L5 and obturator nerve on the superior border of sacral ala to the sacroiliac joint were 20.24?1.20mm,23.80?1.43mm,and 16.26?2.07 mm respectively. There was no statistically significant difference in stability between our method and the traditional fixation by posterior percutaneous sacroiliac screws,though ours seemed better.Follow-ups for the 17 cases averaged 2.2 years,re- vealing fine functional recovery in all according to Matta scoring.Conclusions Euthyphoria reduction and per- cutaneous cannulated screw fixation can lead to sufficient biomechanical stability for the sacroiliac joint and effec- tively avoid nervous injuries.In addition,our method is simple and clinically effective,It is recommendable for small and middle-sized hospitals.

Objective To determine the activities of 131I for treating differentiated thyroid carcinoma with diffuse pulmonary metastases ( DTC-DPM ) from the perspective of internal radiation dosimetry.Methods According to Medical Internal Radiation Dosimetry (MIRD) schema, the activity constraint,from which the whole bdy retention at 48 h should not exceed 2.96 GBq (2.96 GBq rule), was converted to dose-rate constraint(DRC) to lungs at 48 h ( DRCLU ·48 h ) in 131I therapy for DTC-DPM. Based on the assumption of DRCLU·48 h at 48 h in lung, the fractions of whole body activities ( F48 ), the effective half times of 131I in lungs ( TLL ) and the remainder of body ( TRB ) were 0.6-0.9, 20- 120 h, and 10- 20 h, respectively. The maximum safe activities of 131I for different human phantoms from the Organ Level Internal Dose Assessment (OLINDA) software were calculated. Results According to MIRD schema and 2.96 GBq rule, DRCLU ·48 h should not exceed 46.4 mGy/h in 131I therapy for DTC-DPM. Depending on varying F48 h,TLL and TRB, the maximum safe activities of 131I were 6.77-81.36, 5.29-56.20, 5.08-55.19 and 3.87-40. 52 GBq for the male adult, female adult, 15-year-old, and 10-year-old patients with DTC-DPM, respec tively. Conclusion Dosimetry-guided 131I therapy for DTC-DPM considers adequately the differences of 131I kinetics in individual patients and can adjust administered activities of 131I on the precondition of avoiding radiological pneumonitis and pulmonary fibrosis.

Objective To explore the role of the DACT2 gene in the occurrence and development of renal cell carcinoma(RCC).Methods The samples of RCC tissues and corresponding tumor-adjacent tissues after radical operation and normal kidney tissues were collected.The methylation specific PCR (MSP) and real time fluorescence reverse transcriptase-PCR (RT-PCR) methods were adopted to detect the methylation status and mRNA expression of DACT2.The streptavidin-peroxidase (SP) method labeled by immunohistochemistry peroxidase was used to examine the expression of β-catenin protein.Then the relationship between DACT2 gene methylation status and mRNA expression with the clinicopathologic characteristics was analyzed.The relationship between DACT2 gene methylation with mRNA and β-catenin expression was analysed,as well.Results The DACT2 mRNA relative expression level in RCC tissues was 0.427±0.025,which was significantly lower than (0.801±0.047) in tumor-adjacent tissues and (0.872±0.022) in normal tissue,the positive rate of DACT2 gene methylation in RCC tissues was 45.76%,which was significantly higher than 6.78% in tumor-adjacent tissues and 5.08% in normal tissues,the difference was statistically significant (P<0.05),while the difference between tumor-adjacent tissues and normal tissues had no statistical significance (P>0.05).The DACT2 gene mRNA expression level in RCC tissues and promoter area methylation occurrence rate had no obvious correlation with the clinical data such as patients age,gender,tumor size,clinical stage and Fuhrman grade (P>0.05).The DACT2 gene mRNA relative level in the methylation group was lower than that in the non-methylation group,the difference was statistically significant (P<0.05).The expression rate of β-catenin protein in cytoplasma in RCC tissues was higher than that in the tumor-adjacent tissues and normal tissues,the difference was statistically significant (P<0.05),moreover,DACT2 gen methylation had a positive correlation with β-catenin protein expression (r=0.324,P=0.012).Conclusion The decrease of DACT2 gene promoter area methylation and mRNA relative expression level may participate in the RCC occurrence,but has no relationship with RCC clinical progression.Methylation occurred in DACT2 gene promoter area may be one of reasons causing mRNA relative expression decrase.DACT2 gene methylation occurrence in RCC tissue might be related to the high expression of β-catenin.

Staphylococcus aureus ( S. aureus ) osteomyelitis, a significant complication for patients un-dergoing fracture fixation, is a great challenge for orthopaedic surgeons due to its extreme difficulty in mangae-ment. Animal models play an important role in exploring the pathogenesis of osteomyelitis and determining the efficacy of prophylactics and therapeutic treatment. To help understand current animal models of S. aureus os-teomyelitis, we conduct a systematic search to identify animal experiments that have investigated the management of S. aureus osteomyelitis. Experimental studies are categorized by animal species and are further classified by the setting of infection. Study methods are summarized and the advantages and disadvantages of each species and model are discussed.

Objective To investigate the expression of metastasis associated gene-1 (mag-1) and mammalian target of rapamycin (mTOR) in rectal cancer, and its correlation with clinical pathology. Methods The expressions of mag-1 and mTOR in 60 rectal cancer tissue, 30 adenoma tissues and 10 normal rectal tissues were detected by immunohistochemical method, and the correlation between the expression levels and rectal cancer clinical pathologic characteristics was discussed. Results The positive expression rates of mag-1 and mTOR in rectal cancer tissue were significantly higher than those in normal rectal tissue and adenoma tissue:55%(33/60) vs. 1/10 and 27%(8/30), 58%(35/60) vs. 2/10 and 30% (9/30), and there were statistical differences (P<0.05). The expression levels of mag-1 and mTOR in rectal cancer tissue were correlated with carcinoembryonic antigen on admission, degree of cell differentiation, TNM stage, lymph node metastasis and distant metastasis (P<0.01 or<0.05), but had no correlation with age, gender, neoplasm location and neoplasm appearance (P > 0.05). The correlation analysis result showed that the expressions of mag-1 and mTOR were positively correlated in rectal cancer tissue (r=0.730, P<0.01). Conclusions The mag-1 and mTOR may correlate with invasion and metastasis in rectal cancer, and monitoring mag-1 and mTOR expression has a certain value for determining biological behavior of rectal cancer.