BACKGROUND:Modern imaging technology, computer-aided design and processing, rapid prototyping technology and other digital technology have enabled a new era of orthopedic field. Digitalization, individualization, minimal invasion, precision and intel igence are the potential topics in future orthopedics. ＜br> OBJECTIVE:To investigate clinical efficacy of computer-aided design of digital plate in the treatment of hip dislocations associated with posterior wal acetabular fracture. ＜br> METHODS:Sixteen patients of posterior wal acetabular fractures associated with hip dislocations were repaired with customized digital plate fixation. According to Thompson-Epstein fracture type:there were 7 cases with typeⅡ, 5 cases with type Ⅲ, 2 cases with type Ⅳ, and 2 cases with type Ⅴ. Radiographic examination showed the presence of 2-5 mm displacement on the acetabular articular surface, average 3 mm. The time from the injury to hospitalization was 6 hours to 2 weeks, average 1.5 days. After admission, patients underwent femoral condyle bone traction, 12 cases achieved a reduction and three cases of femoral head entrapment were reset during surgery. At 4-10 days after admission, al patients received computer-aided design of digital plate fixation for acetabular fractures. ＜br> RESULTS AND CONCLUSION:Postoperative fracture quality was assessed according to Matta standards, 15 cases had anatomical reduction (displacement＜1 mm) and 1 case had poor reduction (displacement 2-3 mm). The findings indicate that, computer-aided design of digital customized plate has achieved individualized and precise outcomes, as wel as firm fixation in the treatment of hip dislocations associated with posterior wal acetabular fracture. It avoids intraoperative repeated shaping, effectively restores the integrity of posterior wal , al ows early functional exercise, and provides alternative internal fixation for hip dislocations associated with posterior wal acetabular fracture.

Summary: The inhibitory effect of niacinamide on tumor necrosis factor-α (TNF-α) induced annulus fibrous (AF) degradation was assessed, and the mechanism of the inhibition was investigated. Chiba's intervertebral disc (IVD) culture model was established. Forty-eight IVDs from 12 adult Japanese white rabbits were randomly divided into 4 groups (12 IVDs in each group), and various concentrations of niacinamide and TNF-α were added to the medium for intervention: negative control group, niacinamide control group (0.5 mg/mL niacinamide), degeneration group (10 ng/mL TNF-α), and treatment group (0.5 mg/mL niacinamide and 10 ng/mL TNF-α). After one week's culture, AFs were collected for glycosaminoglycan (GS) content measurement, safranin O-fast green staining, and immunohistochemical staining for typeⅠ,Ⅱ collagen and cysteine containing aspartate specific protease-3 (Caspase-3). It was found that the GS content in treatment group was increased by about 48% as compared with degeneration group (t=16.93, P<0.001), and close to that in niacinamide control group (r=0.71, P=0.667). Safranine O-fast green staining exhibited higher staining density and better histological structure of AF in the treatment group as compared with the degeneration group. Immunohistochemical staining for both Type Ⅰ and Ⅱ collagen demonstrated that lameilar structure and continuity of collagen in treatment group were better reserved than in degeneration group. Positive staining rate of Caspase-3 in AFs of negative control group, niacinamide control group, degeneration group and treatment group was 3.4%, 4.3%, 17.9% and 10.3% respectively. The positive rate in treatment group was significantly lower than in degeneration group (P<0.01). It was concluded that niacinamide could effectively alleviate TNF-α induced destruction and synthesis inhibition of matrix ingredients in AFs. The inhibition may be related with reduction of expression of Caspase-3. Thus, niacinamide is of potential for IVD degeneration clinical treatment.

Objective:To explore the effects of K-ras gene on the expressions of oncogenes and cancer suppressor genes in human bronchial epithelial (HBE) cells which were exposed to PM 2.5. Methods:According to the mRNA sequence of K-ras gene provided by GenBank in September 2019, interference sequences were designed and synthesized, and the recombinant lentiviral vector was transfected into HBE cell to construct the K-ras gene-silenced cells. HBE cells and K-ras gene-silenced cells were exposed to 10 μg/ml, 50 μg/ml PM 2.5 suspension and 10 μmol/L Cr 6+. Real-time fluorescent quantitative PCR was used to detect the mRNA expression levels of c-myc, c-fos, N-ras, cyclin-D1, p16 and p53 genes, the expression levels of p53 and c-myc proteins were detected by Western blot. Results:In K-ras silenced cell group, K-ras mRNA expression level decreased (80.5%±3.6%) and K-ras protein level decreased (58.9%±4.7%) when compared with the control group ( P<0.01) . Compared with the correspoding cell control group without exposure, the mRNA expression levels of c-myc, c-fos, N-ras and cyclin-D1 genes in HBE cell group exposed to different concentrations of PM 2.5, K-ras silenced cell group exposed to different concentrations of PM 2.5, HBE cell group exposed to 10 μmol/L Cr 6+ and K-ras silenced cell group exposed to 10 μmol/L Cr 6+ were increased, the mRNA expressions of p16 and p53 genes were decreased ( P<0.01) . Compared with HBE cell group exposed to 10 μg/ml PM 2.5, the mRNA expressions of c-myc, c-fos and p16 genes in K-ras silenced cells exposed to 10 μg/ml PM 2.5 were decreased, and the p53 mRNA level was increased ( P<0.01) . Compared with HBE cell group exposed to 50 μg/ml PM 2.5, the mRNA expression levels of c-fos, N-ras, cyclin-D1, p16 and p53 genes in K-ras silenced cell group exposed to 50 μg/ml PM 2.5 were decreased ( P<0.01) . Compared with the HBE cell group without exposure, c-myc protein increased and p53 protein decreased in HBE cells exposed to 50 μg/ml PM 2.5 ( P<0.05) . Compared with the K-ras silenced cell group without exposure, c-myc protein increased in K-ras silenced cells exposed to 50 μg/ml PM 2.5 ( P<0.05) . Conclusion:PM 2.5 can increase the expression levels of oncogenes in HBE cells, and K-ras gene silencing can inhibit the expression levels of oncogenes in HBE cells treated with PM 2.5.

Objective:To explore the effects of K-ras gene on the expressions of oncogenes and cancer suppressor genes in human bronchial epithelial (HBE) cells which were exposed to PM 2.5. Methods:According to the mRNA sequence of K-ras gene provided by GenBank in September 2019, interference sequences were designed and synthesized, and the recombinant lentiviral vector was transfected into HBE cell to construct the K-ras gene-silenced cells. HBE cells and K-ras gene-silenced cells were exposed to 10 μg/ml, 50 μg/ml PM 2.5 suspension and 10 μmol/L Cr 6+. Real-time fluorescent quantitative PCR was used to detect the mRNA expression levels of c-myc, c-fos, N-ras, cyclin-D1, p16 and p53 genes, the expression levels of p53 and c-myc proteins were detected by Western blot. Results:In K-ras silenced cell group, K-ras mRNA expression level decreased (80.5%±3.6%) and K-ras protein level decreased (58.9%±4.7%) when compared with the control group ( P<0.01) . Compared with the correspoding cell control group without exposure, the mRNA expression levels of c-myc, c-fos, N-ras and cyclin-D1 genes in HBE cell group exposed to different concentrations of PM 2.5, K-ras silenced cell group exposed to different concentrations of PM 2.5, HBE cell group exposed to 10 μmol/L Cr 6+ and K-ras silenced cell group exposed to 10 μmol/L Cr 6+ were increased, the mRNA expressions of p16 and p53 genes were decreased ( P<0.01) . Compared with HBE cell group exposed to 10 μg/ml PM 2.5, the mRNA expressions of c-myc, c-fos and p16 genes in K-ras silenced cells exposed to 10 μg/ml PM 2.5 were decreased, and the p53 mRNA level was increased ( P<0.01) . Compared with HBE cell group exposed to 50 μg/ml PM 2.5, the mRNA expression levels of c-fos, N-ras, cyclin-D1, p16 and p53 genes in K-ras silenced cell group exposed to 50 μg/ml PM 2.5 were decreased ( P<0.01) . Compared with the HBE cell group without exposure, c-myc protein increased and p53 protein decreased in HBE cells exposed to 50 μg/ml PM 2.5 ( P<0.05) . Compared with the K-ras silenced cell group without exposure, c-myc protein increased in K-ras silenced cells exposed to 50 μg/ml PM 2.5 ( P<0.05) . Conclusion:PM 2.5 can increase the expression levels of oncogenes in HBE cells, and K-ras gene silencing can inhibit the expression levels of oncogenes in HBE cells treated with PM 2.5.

Objective:To construct the c-myc gene silenced hepatocytes, study the effect of c-myc gene silence on expression of oncogenes and apoptosis genes in hepatocytes treated with PM2.5.Methods:According to the c-myc gene mRNA sequence provided by GenBank, three interfering sequences were designed and synthesized, the recombinant lentiviral vector was transfected into L02 hepatocytes. The real-time quantitative PCR and western blotting were used to identify the effect of c-myc gene silencing. L02 cells and c-myc gene silenced cells were used as experimental subjects. The normal L02 cells and c-myc silenced cells were treated with 50 μg/ml PM 2.5 water soluble solution, 10 μM positive control Cr 6+ and a blank control, the treatment period was 24 h. The mRNA levels of oncogenes (c-myc, c-fos, k-ras, p53) and apoptotic genes (Caspase-3, Caspase-8, Caspase-9) were detected by real-time PCR. The protein levels of oncogenes and apoptotic genes were detected by western blotting. Results:The mRNA level and protein level of c-myc decreased by 81% and 70% in c-myc silenced cells when compared with the normal L02 hepatocytes, the above results indicate that c-myc gene silenced cells were successfully constructed. After c-myc silenced cells were treated with PM2.5 water soluble solution, The mRNA levels of c-myc, c-fos, and k-ras decreased by 84.1%, 45.4%, and 54.6% ( P<0.05) , p53 increased by 192.9% ( P<0.05) , and the expression of Caspase-3, Caspase-8, and Caspase-9 decreased by 24.4%, 36.1%, 60.9% ( P<0.05) . In the Cr 6+ positive control group, the expression of c-myc, c-fos, and k-ras decreased by 72.1%, 82.2%, and 54.0% ( P<0.05) , p53 increased by 250.0% ( P<0.05) , the expression of Caspase-3, Caspase-8, and Caspase-9 decreased by 34.6%, 36.0%, 68.9% ( P<0.05) , respectively, when compared with the normal L02 hepatocytes ( P<0.05) . Western blotting results showed that the protein levels of c-myc and c-fos increased, p53 decreased after PM 2.5 exposure; the protein levels of Caspase-3, Caspase-8, Caspase-9 increased after PM 2.5 exposure ( P<0.05) . When in comparison with the c-myc silenced group, the protein levels of c-myc and c-fos decreased, p53 protein increased in PM 2.5 exposed group ( P<0.05) . Conclusion:c-myc gene silenced cells were successfully constructed in this paper. PM 2.5 could promote the expression of oncogenes and apoptotic genes in L02 cells, and c-myc gene silencing can inhibit the expression of oncogenes and apoptotic genes after PM 2.5 treatment in L02 cells.

Objective:To study the effect of p38 mitogen-activated protein kinase (MAPK) gene silencing on expression of apoptotic genes and oncogenes in hepatocytes treated with PM 2.5. Methods:From June to September 2019, according to the p38MAPK gene mRNA sequence provided by GenBank, three interfering sequences were designed and synthesized, ligated into PLVX-shRNA2-puro after annealing, and the recombinant lentiviral vector was transfected into L02 hepatocytes. The p38MAPK silencing cells were identified by real-time fluorescent quantitative PCR and western blotting. The normal L02 cells and p38MAPK silencing cells were treated with 50 μg/mL PM 2.5 water soluble solution, 10 μmol/L positive control Cr 6+, and a blank control group was set up, the treatment time was 24 h. The mRNA levels of oncogenes (c-fos, c-myc, k-ras) , tumor suppressor gene (p53) and apoptotic genes (Caspase-3, Caspase-8, Caspase-9) were detected by real-time PCR. The protein levels of oncogenes and apoptotic genes were detected by Western blotting. Results:The expression levels of p38MAPK mRNA and protein in p38MAPK gene silencing cells were significantly lower than those in L02 hepatocytes ( P<0.05) , and the p38MAPK gene silencing cell line was successfully constructed. Compared with the blank control group, the expression levels of the oncogenes c-fos, c-myc, k-ras and the apoptosis genes Caspase-3, Caspase-8 and Caspase-9 increased, the expression level of tumor suppressor gene p53 decreased in the L02 hepatocyte group treated with PM 2.5 water soluble matter, and the differences were statistically significant ( P<0.05) . Compared with the L02 hepatocytes group treated with PM 2.5 water soluble matter, the expression levels of the oncogenes c-fos, c-myc, k-ras and apoptosis genes Caspase-3, Caspase-8 and Caspase-9 decreased, the expression level of tumor suppressor gene p53 increased in the p38MAPK gene silencing cells group treated with PM 2.5 water soluble matter, and the differences were statistically significant ( P<0.05) . Conclusion:PM 2.5 has effects on the expression of oncogenes, tumor suppressor genes and apoptotic genes in L02 hepatocytes, while p38MAPK gene silencing can inhibit the effects of PM 2.5 on L02 hepatocytes.

Objective:To construct the c-myc gene silenced hepatocytes, study the effect of c-myc gene silence on expression of oncogenes and apoptosis genes in hepatocytes treated with PM2.5.Methods:According to the c-myc gene mRNA sequence provided by GenBank, three interfering sequences were designed and synthesized, the recombinant lentiviral vector was transfected into L02 hepatocytes. The real-time quantitative PCR and western blotting were used to identify the effect of c-myc gene silencing. L02 cells and c-myc gene silenced cells were used as experimental subjects. The normal L02 cells and c-myc silenced cells were treated with 50 μg/ml PM 2.5 water soluble solution, 10 μM positive control Cr 6+ and a blank control, the treatment period was 24 h. The mRNA levels of oncogenes (c-myc, c-fos, k-ras, p53) and apoptotic genes (Caspase-3, Caspase-8, Caspase-9) were detected by real-time PCR. The protein levels of oncogenes and apoptotic genes were detected by western blotting. Results:The mRNA level and protein level of c-myc decreased by 81% and 70% in c-myc silenced cells when compared with the normal L02 hepatocytes, the above results indicate that c-myc gene silenced cells were successfully constructed. After c-myc silenced cells were treated with PM2.5 water soluble solution, The mRNA levels of c-myc, c-fos, and k-ras decreased by 84.1%, 45.4%, and 54.6% ( P<0.05) , p53 increased by 192.9% ( P<0.05) , and the expression of Caspase-3, Caspase-8, and Caspase-9 decreased by 24.4%, 36.1%, 60.9% ( P<0.05) . In the Cr 6+ positive control group, the expression of c-myc, c-fos, and k-ras decreased by 72.1%, 82.2%, and 54.0% ( P<0.05) , p53 increased by 250.0% ( P<0.05) , the expression of Caspase-3, Caspase-8, and Caspase-9 decreased by 34.6%, 36.0%, 68.9% ( P<0.05) , respectively, when compared with the normal L02 hepatocytes ( P<0.05) . Western blotting results showed that the protein levels of c-myc and c-fos increased, p53 decreased after PM 2.5 exposure; the protein levels of Caspase-3, Caspase-8, Caspase-9 increased after PM 2.5 exposure ( P<0.05) . When in comparison with the c-myc silenced group, the protein levels of c-myc and c-fos decreased, p53 protein increased in PM 2.5 exposed group ( P<0.05) . Conclusion:c-myc gene silenced cells were successfully constructed in this paper. PM 2.5 could promote the expression of oncogenes and apoptotic genes in L02 cells, and c-myc gene silencing can inhibit the expression of oncogenes and apoptotic genes after PM 2.5 treatment in L02 cells.

Objective:To study the effect of p38 mitogen-activated protein kinase (MAPK) gene silencing on expression of apoptotic genes and oncogenes in hepatocytes treated with PM 2.5. Methods:From June to September 2019, according to the p38MAPK gene mRNA sequence provided by GenBank, three interfering sequences were designed and synthesized, ligated into PLVX-shRNA2-puro after annealing, and the recombinant lentiviral vector was transfected into L02 hepatocytes. The p38MAPK silencing cells were identified by real-time fluorescent quantitative PCR and western blotting. The normal L02 cells and p38MAPK silencing cells were treated with 50 μg/mL PM 2.5 water soluble solution, 10 μmol/L positive control Cr 6+, and a blank control group was set up, the treatment time was 24 h. The mRNA levels of oncogenes (c-fos, c-myc, k-ras) , tumor suppressor gene (p53) and apoptotic genes (Caspase-3, Caspase-8, Caspase-9) were detected by real-time PCR. The protein levels of oncogenes and apoptotic genes were detected by Western blotting. Results:The expression levels of p38MAPK mRNA and protein in p38MAPK gene silencing cells were significantly lower than those in L02 hepatocytes ( P<0.05) , and the p38MAPK gene silencing cell line was successfully constructed. Compared with the blank control group, the expression levels of the oncogenes c-fos, c-myc, k-ras and the apoptosis genes Caspase-3, Caspase-8 and Caspase-9 increased, the expression level of tumor suppressor gene p53 decreased in the L02 hepatocyte group treated with PM 2.5 water soluble matter, and the differences were statistically significant ( P<0.05) . Compared with the L02 hepatocytes group treated with PM 2.5 water soluble matter, the expression levels of the oncogenes c-fos, c-myc, k-ras and apoptosis genes Caspase-3, Caspase-8 and Caspase-9 decreased, the expression level of tumor suppressor gene p53 increased in the p38MAPK gene silencing cells group treated with PM 2.5 water soluble matter, and the differences were statistically significant ( P<0.05) . Conclusion:PM 2.5 has effects on the expression of oncogenes, tumor suppressor genes and apoptotic genes in L02 hepatocytes, while p38MAPK gene silencing can inhibit the effects of PM 2.5 on L02 hepatocytes.