Objective:To explore if there is premature senescence of chondrocytes in patients with Kashin-Beck disease (KBD) and osteoarthritis.Methods:Five knee cartilage samples of KBD, osteoarthritis and control groups were collected, respectively, from the Second Affiliated Hospital of Xi'an Jiaotong University. DNA was then extracted from cartilage samples and DNA methylation was analyzed by Illumina Infinium HumanMethylation450 BeadChip. At the same time, based on genome-wide methylation data, the online DNA methylation aging clock calculator (https://dnamage.genetics.ucla.edu/home) was used to calculate the DNA methylation age (DNAm age) of samples, and the results were compared with their actual ages.Results:In the comparison between KBD group and control group, 1 212 differentially methylated CpG sites were found, including 497 hypermethylated CpG sites and 715 hypomethylated CpG sites, corresponding to 264 hypermethylated genes and 368 hypomethylated genes, respectively. In the comparison between osteoarthritis group and control group, 656 differentially methylated CpG sites were found, including 343 hypermethylated CpG sites and 313 hypomethylated CpG sites, corresponding to 177 hypermethylated genes and 174 hypomethylated genes, respectively. In the above comparison, 367 overlapped CpG sites (corresponding to 182 genes) were found, which were differentially methylated in both KBD and control groups and osteoarthritis and control groups. The results of DNA methylation aging clock showed that the average age acceleration differences between DNAm age and actual age of KBD, osteoarthritis and control groups were 2.549, 0.017, and - 5.364 years, respectively, the DNAm ages of KBD and osteoarthritis groups were greater than the actual ages.Conclusion:The chondrocytes show premature senescence in both KBD and osteoarthritis.

Objective To study the epidemiology of mycoplasma pneumonia (MP) infection in children and its relationship to meteorological conditions in Guilin. Methods Total of 1 342 hospitalized children with respiratory tract infections from Janu-ary 2011 to December 2012 were recruited, and the serum levels of mycoplasma pneumoniae antibody Ig (MP-Ig) were measured. Meanwhile, monthly meteorological conditions were collected. The relationships between the MP-Ig detection rates and meteoro-logical conditions were analyzed by multiple linear regression analysis. Results The total detection rate of MP-Ig was 23.2%with positive rates as 5.51%, 28.99%, 37.31%and 44.97%for 0-1, 1-3, 3-6 and>6 year-old groups respectively (P<0.01). The positive rate of female was higher than that of male with signiifcant difference (30.08%vs 19.05%, P<0.01). The positive rate of spring, summer, autumn and winter were 18.89%, 17.39%, 23.83%and 33.05%individually (P<0.01). Negative correlation was found be-tween detection rate of MP-Ig and monthly mean air temperature (P=0.001). Conclusion MP is one of the commonest pathogens of respiratory tract infection in children and mostly seen in elder female children with a higher incidence in winter. The main me-teorological factor is the low temperature that affects MP prevalence.

Objective To detect the metabolites of Fusarium poae strain isolated from self-produced grain in a new-onset patient with Kaschin-Beck disease,and to provide a scientific basis for studying the relationship between mycotoxins and Kaschin-Beck disease.Methods The strain of Fusarium poae was isolated from the self-produced spike wheat in a family of patient with new-onset Kaschin-Beck disease in Guide County,Qinghai Province in 2016.The isolated Fusarium poae strain was inoculated into potato dextrose agar (PDA) medium for enrichment culture.The metabolites of the fermentation were separated and purified,and the mycotoxins in the metabolite were detected by liquid chromatography-mass spectrometry (LC-MS).The full-spectrum identification was carried out by using the network database Metlin.Results The metabolites of this strain were identified by LC-MS and Metlin full spectrum,and mycotoxins such as trichothecenes,Fumonisin and Zearalenone (ZEA) were detected.A total of 1 601 material characteristic peaks were detected in positive ion mode,the most abundant mycotoxins were Fumonisin FP3,A2,FP1;and a total of 1 000 material characteristic peaks were detected in the negative ion mode,the most abundant were ZEA and monoacetoxyscirpenol.Conclusion The isolated metabolites of Fusarium poae strain mainly contain the three major classes of mycotoxins including trichothecenes,Fumonisins and ZEA.

Objective:To investigate the role of a disintegrin and metalloprotease 12 (ADAM12) gene in chondrocyte injury in patients with Kashin-Beck disease (KBD) and its impact on genes related to insulin-like growth factor binding protein (IGFBP).Methods:Articular cartilage samples were obtained from 5 patients with KBD and 5 control subjects admitted to Honghui Hospital Affiliated to Xi'an Jiaotong University. Chondrocytes were extracted and cultured in vitro. Quantitative real-time PCR (qRT-PCR) and Western blotting were used to detect the expression levels of ADAM12 mRNA and protein in chondrocytes of patients with KBD and control subjects, respectively. Subsequently, ADAM12 gene overexpression was performed using lentivirus in chondrocytes of patients with KBD. MTT assay was used to detect changes in cell viability after ADAM12 gene overexpression, and qRT-PCR was used to detect the mRNA expression levels of chondrocyte differentiation related genes SRY-box transcription factor 9 (SOX9) and type Ⅱ collagen (COLⅡ), apoptosis-related gene B-cell lymphoma/leukaemia-2-associated X protein (BAX), and anabolic related genes IGFBP3 and IGFBP5. Results:The expression levels of ADAM12 mRNA and protein in chondrocytes of patients with KBD (0.57 ± 0.05, 0.81 ± 0.07) were significantly lower than those of control subjects (1.00 ± 0.00, 1.00 ± 0.00), and the differences were statistically significant ( t = - 24.50, - 3.61, P < 0.05). The results of MTT assay showed that the cell viability of chondrocytes in ADAM12 overexpression group (1.09 ± 0.05) was higher than that in empty vector control group (1.00 ± 0.08), and the difference was statistically significant ( t = 4.12, P = 0.031). The results of qRT-PCR showed that compared with empty vector control group, the mRNA expression levels of IGFBP3 (2.35 ± 0.79 vs 0.96 ± 0.25), IGFBP5 (2.13 ± 0.30 vs 0.98 ± 0.34), SOX9 (2.92 ± 0.51 vs 0.94 ± 0.36) and COLⅡ (6.45 ± 2.81 vs 0.87 ± 0.19) in ADAM12 overexpression group were significantly increased, and the differences were statistically significant ( t = 3.19, 5.16, 6.27, 4.10, P < 0.05); while the expression level of BAX mRNA (0.31 ± 0.06 vs 1.02 ± 0.22) was significantly decreased, and the difference was statistically significant ( t = - 11.16, P < 0.001). Conclusion:The ADAM12 gene may have a role in inhibiting apoptosis and promoting differentiation in chondrocyte injury in patients with KBD, and its overexpression can increase expression of IGFBP3 and IGFBP5.

Objective:To investigate the impact of matrix metalloproteinase 13 (MMP13) and low-density lipoprotein receptor-related protein 1 (LRP1) on autophagy of articular chondrocytes in patients with Kashin-Beck disease (KBD).Methods:Human articular cartilage samples obtained from 4 KBD patients and 4 control subjects were collected from Shaanxi Institute for Endemic Disease Prevention and Control, and the expression levels of MMP13 and LRP1 in cartilage tissue were determined using immunohistochemistry (IHC). Chondrocytes were extracted and cultured in vitro, the mRNA and protein expression levels of LRP1 and the autophagy related genes [Beclin 1 (BECN1), microtubule associated protein 1 light chain 3 (LC3)], cartilage injury related genes [MMP13, caspase-3 (CASP3)], chondrocyte differentiation related genes [collagen type Ⅱ alpha 1 chain (COL2A1), and SRY-box transcription factor 9 (SOX9)] were detected by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot (WB), respectively. Chondrocytes from 3 KBD patients were extracted, and MMP13 gene silencing experiment was performed by RNA interference (RNAi) technology, the mRNA and protein expression levels of the above genes were detected by qRT-PCR and WB, respectively. In addition, the antagonist receptor associated protein (RAP) of LRP1 was used to block the LRP1 of human normal chondrocytes (C28/I2 cells), and qRT-PCR and WB were used to detect the mRNA and protein expression levels of LRP1, chondrocyte autophagy, differentiation and cartilage injury related genes, respectively. Results:The IHC results showed that the expression levels of MMP13 (1.67 ± 0.21, 0.59 ± 0.15, 0.51 ± 0.12) in the surface, middle, and deep layers of cartilage tissue of KBD patients were significantly higher than those of control subjects (0.25 ± 0.03, 0.26 ± 0.04, 0.06 ± 0.01), and the differences were statistically significant ( t = - 11.38, P < 0.001; t = - 3.82, - 6.26, P = 0.019, 0.003). The expression levels of LRP1 (0.10 ± 0.02, 0.03 ± 0.01, 0.17 ± 0.03) were significantly lower than those of control subjects (1.63 ± 0.40, 0.44 ± 0.12, 0.34 ± 0.08), and the differences were statistically significant ( t = 6.61, 5.61, 3.64, P = 0.003, 0.005, 0.022). The mRNA and protein expression levels of MMP13, CASP3, SOX9 in chondrocytes of KBD patients were significantly higher than those of control subjects, and the differences were statistically significant ( P < 0.05). The mRNA expression levels of LRP1, LC3, COL2A1 were significantly lower than those of control subjects, and the differences were statistically significant ( P < 0.05). After silencing the MMP13 gene in chondrocytes of KBD patients, there were no significant differences in the mRNA and protein expression levels of LRP1, BECN1, LC3, CASP3, COL2A1, and SOX9 ( P > 0.05). After blocking LRP1 with RAP, the protein expression levels of LRP1, BECN1, LC3, MMP13, COL2A1 and SOX9 in chondrocytes were significantly lower than those in control group ( P < 0.05). Conclusions:There is no direct correlation between MMP13 and abnormal autophagy of articular chondrocytes in KBD patients. After blocking LRP1, the expression of the autophagy related genes BECN1 and LC3 in chondrocytes is decreased.

Objective Aiming at solving the problems of pain on the anteromedial tibia, tibial component loosening and osteoarthritis progression after unicompartmental knee arthroplasty (UKA), the influence of different geometric shapes of tibial component pegs on stress distributions in tibia was analyzed by finite element method. Methods The finite element models with UKA were established and validated. Geometric shapes of tibial component were designed. Under the same loading condition, the tibial components with double-peg, single-keel, double-keel and cross-star were studied for finite element analysis and compared with intact model, so as to evaluate the influence of tibial component with different shapes on stresses of cortical bone in anteromedial tibia, cancellous bone under tibial component and cartilage in contralateral tibia. Results Compared with the intact model, the peak stress of cortical bone in anteromedial tibia with double-peg, single-keel, double-keel and cross-star tibial components increased by 56.1%, 55.9%, 54.5% and 68-4%, respectively. The peak stress of cancellous bone under tibial component with single-keel and double-keels decreased by 8.1% and 15.6% respectively, while the peak stress of cancellous bone under tibial component with double-peg and cross-star increased by 67-9% and 121-5%, which were higher than the fatigue yield stress of cancellous bone. The peak stress of cartilage in contralateral tibia with double-peg, single-keel, double-keel and cross-star tibial components decreased by 42.1%, 26.6%, 24.2% and 28.5%, respectively. ConclusionsThe load distribution of the medial and lateral tibia changed after UKA operation, and a greater load was observed on the replacement side. Single-keel and double-keel tibial components were more effective in reducing stresses on cortical bone in anteromedial tibia and cancellous bone, while the stress distribution in tibia with single-keel tibial component was closer to that of the intact tibia. The research findings can provide theoretical references for designing single-keel tibial component of unicompartmental knee prosthesis which conforms to better mechanical properties of the knee joint.