Objective To investigate the effects of aggrecanase-2 knockdown in chondrocytes from rheumatoid arthritis patient by shRNA infection.Methods Cartilage harvested from rheumatoid arthritis patients who underwent total knee arthroplasty was digested by pancreatin and type Ⅱ collagen enzyme to obtain chondrocytes.Then chondrocytes were cultured and passaged to second or third generation.After aggrecanase-2 shRNA5,aggrecanase-2 shRNA6,aggrecanase-2 shRNA7,aggrecanase-2 shRNA8 infection,growth and morphological changes of the chondrocytes were examined.To select the best target sequence,mRNA expression of aggrecanase-2 and aggrecan was detected by RT-qPCR assay on day 2,5,10,represented with Ct (Cycle threshold) value.Expression of aggrecan protein was detected by immunocytochemistry,represented with IOD (integral optical density) value.Results aggrecanase-2 knockdown had no obvious effects on the morphology and growth of the chondrocytes.MOI (multiplicity of infection) was 100,50,25,and infection efficiency was 90％,60％,30％ with the corresponding viral load of 200 μl,100 μl,50 μl after transfection.mRNA expression of aggrecanase-2 was suppressed significantly,especially the group of shRNA5 which suppressed aggrecanase-2 expression from 0.876 3±0.115 6 to 0.069 9±0.015 1 (P ＜ 0.05).mRNA and protein expression of aggrecan were significantly upregulated after infection.mRNA expression of aggrecan increased from 0.992 1±0.201 3 to 3.049 2±0.278 2 (P ＜ 0.05) and protein expression of aggrecan increased from 496.160 5± 225.673 7 to 4 525.433 0±1 131.813 0 (P ＜ 0.05).Conclusion aggrecanase-2 suppression in chondrocytes by lentivirius infection is an effective method to protect the expression of aggrecan.

Objective To investigate the relationship between blood vessel invasion (BVI) and clinicopathologic features and prognosis in patients with gastric cancer,and analyze related factors affecting the prognosis of patients.Methods The clinicopathological data of 206 patients with gastric cancer who were admitted to the Nanjing Hospital Affiliated to Nanjing Medical University from January 2007 to December 2010 were retrospectively analyzed.The BVI of surgical tumor specimens in all patients was detected by immunohistochemical staininng.All the patients were followed up via outpatient examination and telephone interview up to March 2014.The count data were analyzed using the chi-square test.The survival curve was drawn by Kaplan-Meier method.The survival analysis and univariate analysis were done using the Log-rank test,and multivariate analysis was done using the COX regression model.Results The BVI rate of 206 patients was 27.67％ (57/206).The BVI rate of tumor tissues,tumor differentiation,perineural invasion,T stage,N stage and TNM stage in all patients with gastric cancer were compared,showing significant differences (x2=14.396,9.569,15.579,43.453,30.732,P ＜ 0.05).After operation,188 patients were followed up for 6.0-60.0 months (median,34.0 months),with the follow-up rate of 91.26％ (188/206).Among 188 patients with follow-up,the median survival time and 5-year cumulative survival rate in patients with BVI and with negative BVI were 32.4 months and 19.6％,40.7 months and 42.0％,respectively,with a significant difference in the survival of patients (x2 =9.364,P ＜ 0.05).The results of univariate analysis showed that the diameter of tumor,tumor differentiation,perineural invasion,BVI,T stage,N stage and TNM stage were factors affecting the prognosis of patients with gastric cancer (x2=9.241,17.486,11.243,9.364,27.666,216.745,49.887,P ＜ 0.05).The results of multivariate analysis showed that the diameter of tumormore than 5 cm,BVI,stage T4,stage N3 and stage Ⅲ were independent risk factors affecting the prognosis of patients with gastric cancer (HR =0.502,0.456,0.052,0.001,0.735; 95％ confidence interval:0.334-0.754,0.289-0.720,0.004-0.664,0.000-0.006,0.159-3.398,P ＜ 0.05).Conclusions BVI in patients with gastric cancer is associated with the progression of tumors.The diameter of tumor more than 5cm,BVI,stage T4,stage N3 and stage Ⅲ are independent risk factors affecting the prognosis of patients with gastric cancer,and BVI may be a predictor of poor prognosis of patients with gastric cancer.

BACKGROUND:A large number of domestic and international documents have confirmed that elevated interleukin-1β is associated with primary frozen shoulder.Interleukin-1B gene polymorphisms can affect the transcription and protein expression of interleukin 1β-related genes,resulting in altered levels of cytokines in vivo,and thus altering the incidence of primary frozen shoulder.Through the study of interleukin-1B gene polymorphism and susceptibility to primary frozen shoulder,this study aimed to explore new breakthroughs in the pathogenesis of primary frozen shoulder from the perspective of molecular biology,and to search for susceptibility genes of primary frozen shoulder. OBJECTIVE:To explore the association between linkage disequilibrium of three gene loci in interleukin-1B gene and susceptibility to primary frozen shoulder. METHODS:A case-control study was conducted.There were two groups in this study.One group consisted of 184 patients with primary frozen shoulder,while the other group included 260 healthy controls.The genotypes of interleukin-1B gene loci-511C/T(rs16944),+3954C/T(rs1143634),and-31C/T(rs1143627)were detected by polymerase chain reaction and restriction fragment length polymorphism.The correlation between the probability of linkage disequilibrium and haplotypes and the risk of primary frozen shoulder disease was compared and analyzed. RESULTS AND CONCLUSION:Unconditional Logistic regression analysis showed that the proportion of CT genotypes at rs1143634 and rs1143627 sites increased significantly in the primary frozen shoulder.Linkage disequilibrium analysis showed that rs16944,rs1143634 and rs1143627 tended to be balanced in the control group(D'value<0.1),while there was a certain degree of linkage disequilibrium at rs1143627 and rs1143634 sites in the primary frozen shoulder group(D'value=0.595).Haplotype TTT increased the risk of primary frozen shoulder by 6.66 times compared with CCT type(TTT,OR=6.66,95%CI=1.59-27.88,P=0.009 7).To conclude,there is a certain degree of linkage disequilibrium between interleukin-1B gene loci rs1143627and rs1143634 in patients with primary frozen shoulder;haplotype TTT formed by these three gene loci may increase the risk of developing primary frozen shoulder.

Objective:To analyze the clinical characteristics and management strategies of the patients who were injured in winter snow sports in parts of Qingdao in order to provide useful information for "300 million people participating in ice and snow sports" .Methods:A retrospective study was conducted to analyze the data of 128 patients who had been admitted to Department of Orthopaedics, Qingdao Central Hospital for skiing injuries from November 2019 to March 2021. The gender, type, location and cause of injury, and treatment method of the patients were statistically analyzed.Results:Of the 128 patients, 72 were males and 56 females with an age of (32.0±12.1) years. Single injury accounted for 97.66% (125/128) while multiple injuries 2.34% (3/128). 46 injuries (35.11%, 46/131) due to single board skiing occurred in 44 patients (34.38%, 44/128); 85 injuries (64.89%, 85/131) due to double board skiing occurred in 84 cases (65.62%, 84/128). As for injury types, there were 44 cases (34.38%, 44/128) of soft tissue injury and 84 fractures (65.62%, 84/128). The top 3 injured locations were the lower leg (24.43%, 32/131), the peri-wrist part (18.32%, 24/131), and the peri-knee part (15.27%, 20/131). The most vulnerable location was, respectively, the wrist joint (26.09%, 12/46) in single board skiing and the calf (31.76%, 27/85) in double board skiing. The top three causes of injury were accidental fall (39.06%, 50/128), being struck by someone or emergency evacuation (14.84%, 19/128), and mismatched equipment models (12.50%, 16/128). The proportion of skiing beginners was as high as 76.56% (98/128). Surgical treatment was performed in 49 cases (38.28%, 49/128), and conservative treatment in 79 cases (61.72%, 79/128).Conclusions:The clinical characteristics of the patients who were injured in winter snow sports in parts of Qingdao are high proportions of skiing beginners, falls and fractures. Special attention needs to be paid to wrist protection in single board skiing and checking the ski disengagement device before double board skiing. Most patients may need only conservative treatment because although a number of patients required surgical treatment, severe multiple injuries were rare.

Objective:To evaluate the efficacy of percutaneous vertebroplasty (PVP) by percutaneous bi-level bilateral puncture in the treatment of type ⅡA acute symptomatic osteoporotic thoracolumbar fracture (ASOTLF) with dense bone bands.Methods:From March 2017 to March 2018, 65 patients (65 vertebrae) with type ⅡA ASOTLF with dense bone bands were treated at Department of Orthopaedic Trauma, Qingdao Central Hospital. They were 25 males and 40 females, with an age of (71.6±8.4) years. The time from injury to operation was (3.5±0.7) d. They were divided into 2 groups according to different treatments. In the observation group of 31 cases (31 vertebrae), PVP was conducted by percutaneous bi-level bilateral puncture; in the control group of 34 cases (34 vertebrae), PVP was conducted by percutaneous single-level bilateral puncture. The 2 groups were compared in terms of operation time, volume of polymethyl methacrylate injected, incidence of bone cement leakage, incidence of vertebral refracture, diffusive distribution of bone cement in the vertebral body, and visual analogue scale (VAS) and Oswestry dysfunction index (ODI) at postoperative 1 day and 6 months.Results:There was no significant difference in the preoperative general data between the 2 groups, showing comparability ( P>0.05). Operations were completed uneventfully in all the 65 patients. All the 65 patients were followed up for (8.2±1.2) months. There was no significant difference between the 2 groups in operation time, incidence of bone cement leakage or incidence of vertebral refracture for each vertebral body injured ( P>0.05). The volume of polymethyl methacrylate injected [(6.64±0.93) mL] and the excellent and good rate of diffusive distribution of bone cement in the vertebral body [87.1% (27/31)] in the observation group were significantly larger than those in the control group [(4.36±0.79) mL and 64.7% (22/34)] ( P<0.05). The VAS scores at postoperative 1 day and 6 months [2 (1, 2) and 1 (1, 2)] and ODIs at postoperative 1 day and 6 months (23.7%±1.6% and 18.8%±1.4%) in the observation group were significantly lower than those in the control group [2 (2, 3) and 2 (2, 2); 26.9%±4.2% and 22.1%±3.3%] ( P<0.05). The VAS scores and ODIs at postoperative 1 day and 6 months in all patients were significantly lower than those before operation ( P<0.05). Compression symptoms of the spinal cord and nerve root were observed in none of the patients. Conclusion:In the treatment of type ⅡA ASOTLF with dense bone bands, PVP by percutaneous bi-level bilateral puncture is more effective than the traditional PVP by percutaneous single-level bilateral puncture, and will not increase bone cement leakage or vertebral refracture.

Objective To explore the genetic characteristics in a Chinese family with autosomal dominant lateral temporal lobe epilepsy (ADLTE) and analyze the correlation between genotype and phenotype. Methods The natural history,clinical data and peripheral blood sample were collected in all patients and two healthy members of this ADLTE family.Whole exon sequence(WES)analysis strategy was used to explore the underlying mutations. Possible causative genetic variation was further confirmed by direct PCR and Sanger sequencing. The genotype-phenotype features were compared with previously reported cases. Results A novel pathogenetic LGI1 frameshift mutation p.T134fs was identified in this study. The clinical phenotype was different from reported. Conclusion This study reports a pathogenic LGI1 mutation in a Chinese ADLTE family for the first time, which suggests that LGI1 is a new genetic abnormality of ADLTE in Chinese.

Background Heavy metals are widely present in the environment, difficult to degrade, and bioaccumulative. Children's physical development are not mature, and exposure to heavy metals which may cause irreversible harm to them. However, the current research conclusions are still inconsistent. Therefore, it is very important to pay attention to the impact of heavy metal pollutants on physical development. Objective To detect the level of heavy metal exposure of school-age children in Laizhou Wan, Shandong Province, and to explore the relationship between heavy metal exposure level and physical development indexes of school-age children. Methods The subjects of this study were 318 children born to pregnant women recruited from Laizhou Wan Birth Cohort in Shandong Province from September 2010 to December 2013. The school-age children at age 7 were followed up from July to September 2019. Their urine samples were collected and the levels of arsenic (As), cadmium (Cd), lead (Pb), and mercury (Hg) in urine were detected by inductively coupled plasma mass spectrometry (ICP-MS). At the same time, the height, weight, body mass index (BMI), waist circumference, and body fat percentage of the children were evaluate to analyze the relationship between the four heavy metals in children's urine and the indicators related to children's physical development. Results Among the 318 school-age children with an age (\begin{document}$\bar x \pm s $\end{document}) of (7.77±0.67) years, the median (P₂₅, P₇₅) levels of As, Cd, Pb, and Hg in urine were 73.98 (44.81, 124.61), 0.40 (0.27, 0.58), 1.20 (0.71, 1.72), and 2.38 (1.32, 4.80) µg·g⁻¹ (of creatinine), respectively. The children’s urinary Cd level was positively correlated with their weight, BMI, and waist circumference (P<0.05), and corresponding regression coefficients (b) and its 95% confidence intervals (CI) were 0.99 (0.14-1.83), 0.43 (0.06-0.81), and 1.35 (0.27-2.42), respectively; while the other heavy metals were not related to the above physical development indicators (P>0.05). After sex stratification, it was found that boys’ urinary Cd level was positively correlated with their height, weight, BMI, and waist circumference (P <0.05) with associated b (95%CI) of 1.73 (0.53-2.93), 2.03 (0.60-3.47), 0.75 (0.11-1.38), and 2.66 (0.85-4.47), respectively; such associations were not found in girls. After further stratification of boys’ BMI according to normal, overweight, and obesity, it was found that a higher urinary Cd level was correlated with an increased risk of obesity in boys (P<0.05), and the associated b (95%CI) was 2.34 (1.02-5.36). Conclusion The level of urinary Cd exposure of boys in Laizhou Wan, Shandong Province is positively correlated with their height, weight, BMI, and waist circumference, and may be related to obesity in boys.