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Objective:]To investigate the role of Arpin protein in bone repair by mediating migration of host bone marrow mesenchymal stem cells (BMSCs) to the bone defect area after transplantation of tissue engineering bone (TEB).Methods:Immunofluorescence was used to observe the expression and relative localization of Arpin and Arp2/3 proteins in BMSCs. Lentiviruses that ware designed to interfere with Arpin expression were constructed to transfect BMSCs for knockdown Arpin expression. Knockdown efficiency was verified by real-time quantitative reverse transcription PCR ( qRT-PCR) and Western blot. According to different levels of Arpin protein expression, experiments were divided into empty vector control group and an Arpin expression inhibition group in vitro and in vivo. In vitro experiments: the cell migration model was established with a migration chamber, then the cells from both groups were seeded on the up chamber, and the number of migrated cells were detected by fluorescence microscopy. Cells from both groups were seeded on six-well plates. Model of wound healing experiment was established and wound healing ratio was examined by microscopy. In vivo experiments: 8-week-old C57BL/6 mice were selected and assigned to empty vector control group and Arpin expression inhibition group according to the random number table, with 6 rats per group. Diaphysis of 2 mm and periosteum in the middle femur were excised to make a large segment of bone defects. Then, TEB was transplanted into the defect area and fixed.Green fluorescein-labeled BMSCs (1 million cells per mouse) from empty vector control group and Arpin expression inhibition group were injected through the tail vein. Number of BMSCs homing to the bone defect area was detected by immunofluorescence staining at day 2 and 7 after operation. At 4 weeks after operation, the femur was taken for a Micro-CT scan to analyze bone mass density(BMD), bone volume density (BV/TV), trabecular spacing (Tb.Sp) and trabecular thickness (Tb.Th). Then, the specimens were stained with pathological HE and MASSON staining to observe the quality of bone formation. Results:Mouse BMSCs expressed Arpin protein, which was located at the cell edge relative to Arp2/3. After transfection of lentivirus, BMSCs expressed green fluorescent protein, and the expression of Arpin gene and protein in Arpin expression inhibition group were decreased compared to empty vector control group ( P<0. 01). BMSCs migration was enhanced in Arpin expression inhibition group compared to empty vector control group [(76.6±6.6) vs. (105.7±6.5)] ( P<0. 01). Wound healing was accelerated in Arpin expression inhibition group compared to empty vector control group [(43.8±0.19)% vs. (62.6±3.2)%]( P<0.01). At day 2 after operation, immunofluorescence results showed no significant difference in cell migration between the two groups and almost no labeled cells migrated. At day 7 after operation, more cells migrated to the transplanted area in Arpin expression inhibition group compared to empty vector control group [(5.7±1.5) vs. (11.3±1.5)] ( P<0.01). At 4 weeks after operation, Micro-CT results showed that Arpin expression inhibition group had better bone formation quality than empty vector control group [BMD: (172.7±6.0)mg/cm 3vs. (140.0±6.0)mg/cm 3, BV/TV: (28.8±1.3)% vs. (23.4±0.9)%, Tb.Sp: (0.33±0.01)μm vs. (0.28±0.01)μm, Tb.Th: (0.11±0.01)μm vs.(0.15±0. 01)μm]( P<0.05). Pathological staining showed there were more new bone tissue in Arpin expression inhibition group ( P<0.01). Conclusion:Silencing Arpin protein expression promotes BMSCs to migrate to the bone defect area and improves bone repair effect.
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OBJECTIVE: To observe the changes of chest imaging and prognosis in pneumoconiosis patients in tin smelting workers. METHODS: Ten pneumoconiosis patients working with tin smelting were examined by chest X-ray,computed tomography( CT) photography and dynamic observation on pulmonary imaging to analyze their characteristics and prognosis. RESULTS: There were mild clinical manifestations and no tuberculosis in these 10 cases of tin smelting pneumoconiosis patients. There was no obvious change on the pulmonary ventilation function change. The high k V X-ray chest observation results showed that the circular shadow was the primary small shadow of the two lungs,that were mostly distributed in the medium and upper lung zones of both lungs. In most cases,we found lung texture distortion,deformation or increase,blurred,hilar shadow increased thickening,lymph node calcification,individualized eggshell. There was no pleural changes and emphysema changes. There was no significant change found in 5-10 years of dynamic observation except for 1 case of increased small shadow. The chest CT examination in 2011 showed 2-5 mm nodular shadows. Among them,we found 7 cases of small nodules from the upper lobe evenly distributed to the middle of the lobe,the lower back lobe of the lungs,lower basal ganglia lesions decreased,lesions were diffuse distribution of the whole lung,and the small nodules in 3 cases. Interval lobular thickening at varying degrees were found in 5 cases,lobular central or apoptotic pulmonary emphysema were found in 4 cases,and 1 case of pulmonary bullae formation was found. The results of chest CT examination in 2016 showed 2 cases of diffuse pulmonary nodules,3 cases of thickening of lobular septum and 2 cases of pulmonary emphysema compared with the CT result in 2011. CONCLUSION: There was no obvious lung small shadow absorption found in tin smelting pneumoconiosis patients after 5 to 10 years of X-ray dynamic observation,and progress of lesions could be seen. CT examination is helpful for follow-up observation in tin smelting pneumoconiosis.
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Objective To study the relationship between cyclooxygenase 2 (COX-2) polymor phisms and the susceptibility of bladder cancer.Methods Polymerase chain reaction restricted frag ments length polymorphism (PCR-RFLP) and the primer introduced restriction analysis (PIRA-PCR)assay were used to genotype the COX-2-765G/C, 1195G/A and 8473T/C polymorphisms in a case control study of 180 bladder cancer cases and 180 cancer free controls in a Chinese population.Re stilts The distribution of the genotype frequencies of 765G/C and 1195G/A were not statisticallydifferent between the cases and controls (P=0.582 for-765G/C and P=0.270 for-1195G/A).Poly morphisms of COX 2-8473T/C were associated with the susceptibility to bladder cancer.The individ uals with the 8473C allele had a decreased risk of bladder cancer (OR=0.56,95% CI=0.35 0.88).Conclusions Polymorphisms of COX-2-765G/C and-1195G/A are not associated with the suscepti bility to bladder cancer.However,COX-2-8473T/C can reduce the risk of bladder cancer.
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Objective To investigate the relationships between genetic polymorphism of CYP2A6 alone or in combination with smoking and hereditary susceptibility to bladder cancer.Methods Based on case-control study,CYP2A6*4 was determined by the nested polymerase chianreaction(nPCR)in 186 patients with bladder cancer and 192 nontumorous controls.The relations between the genetypes of CYP2A6*4 alone or combinated with smoking and bladder cancer was estimated with the X2 test and logistic regression model.Results In the case subjects,the number of the wil/wil genetype was 168,the number of the wil/del genetype was 13,and the number of the del/del genetype was 5.In the control subjects,the number of the wil/wil genetype was 150,the number of the wil/del genetype was 32,and the number of the del/del genetype was 10.The frequency of CYP2A6 del allele was significantly lower in the case Subjects(9.68%)than the controls(21.88%,P＜0.05,OR:0.383).When eombinated with smoking,the risk of bladder cancer in smokers was significantly higher than nonsmoker(P＜0.05,OR=2.322).In smokers,the frequency of CYP2A6 del allele was significantly lower in cases(7.88%)than controls(28.00%,P＜0.05,OR=0.221).In smoking people,the one with CYP2A6 del genotype had a lower risk of bladder cancer than the one with CYP2A6 wild genotype(OR=0.221,95%CI:0.092,0.534).Conclusions Genetic polymorphisms of CYP2A6 are associated with the susceptibility to bladder cancer and have interaction with smoking in carcinogenesis of bladder cancer.Deficient CYP2A6 activity to genetic polymorphism mayreduee bladder cancer risk in smokers.