BACKGROUND:WIF-1 is a tumor suppressor gene. Promoter hypermethylation causes WIF-1 down-regulationin most tumors. DNA methylation inhibitor can lead to gene demethylation and restore its expression. OBJECTIVE:To observe the differences of tumor pathology and, WIF-1 mRNAand proteinchanges using WIF-1 or 5-aza-2'-deoxycytidine demethylation in animalmodels of osteosarcoma.
METHODS:Murine osteosarcoma models were established and divided into three groups. In the control group, no treatment was given. In the 5-aza-2'-deoxycytidine group, an appropriate amount of 5-aza-2'-deoxycytidine was injected ineach mouse daily. In the WIF-1 group, an appropriate amount of Wnt/β-catenin signal transduction pathway inhibitor WIF-1 was injected in each mouse daily. Seven days after medication, the weight of nude mouse was weighed every 7 days. Short tumor diameter (a) and the long diameter (b) were measured. Therelative tumor volume was calculated. The relative growth rate of tumor was calculated at 7, 14, 21, 28 and 56 days. Four nude mice from ach group were sacrificed by puling the neck at 7, 14, 21, 28 and 56 days after medication. Tumor tissues were stripped and the weight of them was weighed. Pathological analysis of the tumor was conducted. The expression of WIF-1protein and WIF-1 mRNA was detected in osteosarcoma at 56 days after medication in the three groups.
RESULTS AND CONCLUSION:(1) Compared withthe medication and control groups, the weight of nude mice was increased at 7, 14, 21, 28 and 56 days in the treatment group. No significant difference was found between the medication and control groups. (2) The tumor size was significantly smaler in themedication group than in the control group. WIF-1 mRNA and WIF-1 protein expression was increased in the medication group compared with the control group to different degrees. (3) Results suggested that WIF-1 gene promoter methylation is one of the mechanisms of the development of osteosarcoma. Use of WIF-1 or 5-aza-2'-deoxycytidine demethylation can inhibit tumor growth in animal models of osteosarcoma.

Objective:To development a new method for sensitive detection of Porphyromanus gingivalis (P.gingivalis) based on magnetic encoding nanospheres and upconversion fluorescent encoding nanospheres.Methods:Magnetic and upconversion fluorescent encoding nanospheres were prepared by sol-gel method respectively,combined the monoclonal antibodies specific to P.gingivalis after modifying the surface of nanospheres.The system was used to detect P.gingivalis from mixed bacteria solution of P.gingivalis,F.nucleatum and S.mutans.Fluorescent microscopy with an external 980 nm near-infrared hght pulse laser,scanning and transmission microscope were used to evaluate the effectiveness of the detection system.Results:Magnetic and upconversion encoding nanospheres had better dispersion,particle size uniformity and homogeneous morphology.Besides,the magnetic encoding nanospheres had good magnetic properties and strong fluorescence intensity.P.gingivalis was captured by magnetic and upconversion encoding nanospheres in a mixed solution of the 3 bacteria with a detection limit of 10 CFU/ml.Conclusion:The method designed in this study can capture P.gingivalis sensitively in a mixed bacteria liquid.

Objective: To establish a mouse model of osteoarthritis (OA) and study the bone microarchitecture and bone metabolism of tibial subchondral bone in early stage of OA. Methods: The mouse model of post-traumatic osteoarthritis (PTOA) with anterior cruciate ligament (ACLT) was established by using c57 mice. The Sham operation group served as the control group. All mice were fed with conventional diet. All mice were sacrificed after 4 weeks. The degeneration of knee joint was observed by HE staining and Safranin O-Fast Green staining. The number of osteoclasts was counted by TRAP staining. Micro CT was used to analyze the quantitative parameters of the microstructure of tibia subchondral bone in mice. Serum levels of bone resorption biomarker CTX I and cartilage degeneration marker CTX II were determined. Results: After ACLT 4 weeks, the average score of OARSI in ACLT group was 3.2, which was higher than that in Sham group, and the joint degeneration occurred in mice, presenting the pathological characteristics of early OA. Compared with the sham operation phase, the total subchondral bone volume (TV) of ACLT group was 4.72 mm³, increased by 13.6%; the bone trabecular resolution (Tb.Sp) was 0.130 and 0.154 mm, respectively, and the ACLT group also increased by 18.8%; the bone volume/tissue volume (BV/TV) was 0.470 and 0.294, respectively, and the ACLT group decreased by 48.9%; the bone trabecular thickness (Tb.Th) was 0.162 and 0.083 mm groups, ACLT decreased by 37.5%. Trap staining showed that the number of osteoclasts per unit volume in ACLT group was 72, which was significantly higher than that in sham operation group. The CTX I of mice in the sham operated ACLT group and sham operated group were 20.9 ng/ml and 18.29 ng/ml, with an increase of 48.9% in the ACLT group; the CTX II of mice in the ACLT group and sham operated group were 35.5 ng/ml and 28.6 ng/ml, with an increase of 24.1% in the ACLT group. Conclusion ACLT Mouse model can successfully construct early OA, which confirms the early loss of osteochondral bone and the pathological changes of osteoclast activation in OA, and provides a new specific target for the treatment of OA.

【Objective】 To investigate the current situation concerning volume control of red blood cells in additive solution produced by blood service in Chongqing, and to lay a foundation for promoting the homogenization of preparation process of red blood cells in additive solution. 【Methods】 A questionnaire was designed to investigate the factors related to the preparation of red blood cells in additive solution. The questionnaire was sent by Chongqing Association of Blood Transfusion via E-mail to 18 blood services in the city, and the collected data was sorted, revised and analyzed by research team. 【Results】 A total of 18 blood services(including 1 blood center + 1 sub-center, 6 central blood stations and 11 central blood banks) returned the questionnaires. The results showed that there were differences among blood services across Chongqing, regarding the centrifugal parameters during preparation, the operation mode and monitoring situation of the capacity control during preparation, and the formulation of the capacity standard of red blood cells in additive solution etc. 【Conclusion】 The preparation process of red blood cells in additive solution, produced by Chongqing blood services, should be further standardized, and the capacity control method of this product in Chongqing should be gradually unified to achieve regional homogeneity and to ensure blood safety.

AIM:Bone morphogenetic protein 7(BMP7)reduces the expression of Yes-related protein 1(YAP1)by down-regulating Ajuba level and decreasing extracellular matrix(ECM)deposition.This study aimed to inves-tigate the influence of these factors on modifying the degree of renal fibrosis in rats with diabetic nephropathy.METH-ODS:Eighteen Sprague-Dawley(SD)rats were randomly divided into three groups:the normal control(NC)group,the diabetes mellitus(DM)group,and the DM group treated with BMP7 overexpressing adeno-associated virus(DM+rAAV-BMP7).Each group consisted of six rats.Diabetic kidney disease(DKD)was established in the DM and DM+rAAV-BMP7 groups by injecting 55 mg/kg streptozotocin(STZ)via the tail vein.NRK-52E cells were divided into three groups:the normal glucose(NG)group,the high glucose(HG)group,and the high glucose group treated with recombinant hu-man BMP7(HG+rhBMP7)group.Pathological changes in renal tissues were observed using hematoxylin and eosin(HE)and Sirius red staining.Immunohistochemical staining was performed to examine the expression sites of Ajuba and YAP1 in the renal cortex.Western blot analysis was conducted to determine the expression levels of BMP7,Ajuba,YAP1,colla-gen type Ⅲ(Col-Ⅲ),and fibronectin(FN)in the rat renal cortex and NRK-52E cells.RT-qPCR was used to measure the mRNA levels of Ajuba and YAP1 in the rat renal cortex.RESULTS:Biochemical indices revealed significantly ele-vated levels of blood glucose,serum creatinine,triglycerides,total cholesterol,and 24-hour urinary protein in the DM group compared to the NC group(P<0.05).In the DM+rAAV-BMP7 group,the levels of serum creatinine,24-hour uri-nary protein,triglycerides,and total cholesterol were lower than those in the DM group(P<0.05).Pathological staining demonstrated that the renal interstitium of the DM group exhibited inflammatory cell infiltration,fibrous tissue,collagen fi-ber deposition,disordered renal tubule arrangement,atrophy,and vacuolar degeneration,which were ameliorated in the DM+rAAV-BMP7 group.Immunohistochemistry revealed that Ajuba and YAP1 were mainly expressed in the cytoplasm and nucleus,with high expression in the cytoplasm of the DM group,which was significantly decreased in the DM+rAAV-BMP7 group.Western blot results indicated that the protein levels of FN,Col-Ⅲ,Ajuba,and YAP1 were up-regulated in the DM and the HG groups(P<0.05),but significantly down-regulated in the DM+rAAV-BMP7 group(P<0.05).RT-qP-CR results demonstrated that the mRNA levels of Ajuba and YAP1 were higher in the DM group and significantly lower in the DM+rAAV-BMP7 group(P<0.05).CONCLUSION:The overexpression of BMP7 can ameliorate renal fibrosis in rats with DKD.This effect is likely mediated by the down-regulation of Ajuba,reduction of YAP1 expression,and subse-quent inhibition of ECM deposition.