Summary Analysis of National Surveillance on Kashin-Beck Disease from 1990 to 2023

Objective To analyze the epidemiological characteristics and epidemic situation of children with Kashin-Beck disease(KBD)in China,and provide the basis for formulating prevention and control measures.Methods Fixed-point monitoring,moving-point monitoring,and full coverage of monitoring were promoted successively from 1990 to 2023.Some children(7-12 years old)underwent clinical and right-hand X-ray examinations every year.According to the KBD diagnosis criteria,clinical and X-ray assessments were used to confirm the diagnosis.Results In 1990,the national KBD detectable rate was 21.01%.X-ray detection decreased to below 10%in 2003 and below 5%in 2007.Between 2010 and 2018,the prevalence of KBD in children was less than 0.4%,which fluctuated at a low level,and has decreased to 0%since 2019.Spatial epidemiological analysis indicated a spatial clustering of adult patients prevalence rate in the KBD areas.Conclusion The evaluation results of the elimination of KBD in China over the last 5 years showed that all villages in the monitored areas have reached the elimination standard.While the adult KBD patients still need for policy consideration and care.

Evaluation of the Sensitivity and Specificity of the New Clinical Diagnostic and Classification Criteria for Kashin-Beck Disease,an Endemic Osteoarthritis,in China

This study aimed to evaluate the sensitivity and specificity of the new clinical diagnostic and classification criteria for Kashin-Beck disease （KBD） using six clinical markers： flexion of the distal part of fingers, deformed fingers, enlarged finger joints, shortened fingers, squat down, and dwarfism. One-third of the total population in Linyou County was sampled by stratified random sampling.

In vitro effects of sodium hyaluronate on the proliferation and the apoptosis in chondrocytes from patients with Kashin-Beck disease and osteoarthritis

Objective：To identify the in vitro effects of sodium hyaluronate（HA） on the proliferation and the apoptosis of chondrocytes from patients with Kashin-Beck disease（KBD） and osteoarthritis（OA）. Methods：Samples of articular cartilages from KBD and OA patients, as well as healthy volunteers（6 subjects in each of the 3 groups） were dissected, digested with collagenase and the cells cultured in monolayers. Chondrocytes from each sample were assigned to an untreated group and two HA-treated groups： H0（no HA）, H100（HA, 0.1 g/L） and H500（HA, 0.5 g/L）. The first passage chondrocytes were used to observe proliferation using the MTT assay, and apoptosis by flow cytometry through Annexin V/PI staining. Results：HA promoted proliferation of chondrocytes in all the three groups, and.in KBD and OA groups, for cells cultured for 4 and 6 days, H500 significantly promoted the cell proliferation. The apoptotic rates of both KBD and OA group chondrocytes were in the order H500 〈 HA100 〈 H0. Conclusion：Sodium hyaluronate administration has a dosedependent in vitro effect to promote proliferation and inhibit apoptosis of chondrocytes from patients with KBD and OA.

Changing Grains for the Prevention and Treatment of Kashin-Beck Disease in Children: a Meta-analysis

To evaluate the efficacy of changing grains on the prevention and treatment of Kashin-Beck Disease （KBD） in children, community-based trials were acquired from seven electronic databases （up to July 2014）. As a result, the methodological quality of the six trials that have been included into our analysis was low. The pooled ORs favoring the prevention and treatment effects of changing grains were 0.15 （95% CI： 0.03-0.70） and 2.13 （95% CI： 1.44-3.16） respectively by meta-analysis. Subgroup analysis demonstrated the pooled OR favoring treatment effect of exchanging grains rather than drying grains both compared with endemic grains. The results showed that changing grains had obvious effects on the prevention and treatment of KBD in children. However, the evidences were limited by the potential biases and confounders. Large and well-designed trials are still needed.

Exploring Genome-wide DNA Methylation Profiles Altered in Kashin-Beck Disease Using Infinium Human Methylation 450 Bead Chips

To understand how differentially methylated genes（DMGs）might affect the pathogenesis of Kashin-Beck disease（KBD）.Genome-wide methylation profiling of whole blood from 12matched KBD and controls pairs was performed using a high-resolution Infinium 450 K methylation array.In total,97 CpG sites were differentially

Serum Metabolomic Indicates Potential Biomarkers and Metabolic Pathways of Pediatric Kashin-Beck Disease

Objective To explore potential serum biomarkers of children with Kashin-Beck Disease(KBD)and the metabolic pathways to which the biomarkers belong.Methods A two-stage metabolomic study was employed.The discovery cohort included 56 patients,51 internal controls,and 50 external controls.The metabolites were determined by HPLC-(Q-TOF)-MS and confirmed by Human Metabolome Databases(HMDB)and Metlin databases.MetaboAnalyst 3.0 and the Kyoto Encyclopedia of Genes and Genomes(KEGG)database were used to analyze the metabolic pathways of the candidate metabolites.The use of HPLC-(Q-TRAP)-MS enabled quantitative detection of the target metabolites which were chosen using the discovery study and verified in another independent verification cohort of 31 patients,41 internal controls,and 50 external controls.Results Eight candidate metabolites were identified out in the discovery study,namely kynurenic acid,N-α-acetylarginine,6-hydroxymelatonin,sphinganine,ceramide,sphingosine-1 P,spermidine,and glycine.These metabolites exist in sphingolipid,glutathione,and tryptophan metabolic pathways.In the second-stage study,five candidate metabolites were validated,including kynurenic acid,N-α-acetylarginine,sphinganine,spermidine,and sphingosine-1 P.Except for spermidine,all substances exhibited low expression in the case group compared with the external control group,and the difference in levels of sphinganine,spermidine,and sphingosine-1 P was statistically significant.Conclusion The direction of change of levels of sphinganine,spermidine,and sphingosine-1 P in the two-stage study cohorts was completely consistent,and the differences were statistically significant.Therefore,these substances can be used as potential biomarkers of KBD.Furthermore,these results raise the possibility that sphingolipid metabolic pathways may be closely related to KBD.

Crosstalk between CpG Methylation and Polymorphisms (CpG-SNPs) in the Promotor Region of DIO2 in Kashin-Beck Disease

Objective This study was designed to determine the methylation profile of four CpGs and the genotypes of two CpG-SNPs located in promoter region of DIO2 in patients with Kashin-Beck disease(KBD).We also analyzed the interaction between the CpGs methylations and CpG-SNPs.Methods Whole blood specimens were collected from 16 KBD patients and 16 healthy subjects.Four CpGs and two CpG-SNPs in the promoter regions of DIO2 were detected using matrix-assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS).The CpGs methylation levels were compared between samples from KBD patients and healthy subjects.The methylation levels were also analyzed in KBD patients with different CpG-SNP genotypes.Results The mRNA expression of DIO2 in whole blood of KBD patients was significnatly lower than in healthy controls(P<0.05).The methylation levels of DIO2-1_CpG_3 in KBD patients were significantly higher than those in healthy controls(P<0.05).The methylation levels of four CpGs were not significantly different between KBD patients and healthy controls.The methylation level of DIO2-1_CpG_3 in the promoter region of DIO2 in KBD patients with GA/AA genotype was significantly higher than that of KBD patients with GG genotype(P<0.05).Conclusion The methylation level of DIO2 increases in KBD patients.Similar trends exist in KBD carriers of variant genotypes of CpG-SNPs DIO2 rs955849187.

Hyaluronic acid inhibited the upregulation of heat-shock protein 70 in human chondrocytes from osteoarthritis and Kashin-Beck disease

This study aimed to investigate the effect of hyaluronic acid(HA)on the expression of heat-shock protein 70(HSP70)in chondrocytes isolated from patients with osteoarthritis(OA)and Kashin-Beck disease(KBD).The chondrocytes were collected from OA and KBD patients,and chondrocytes isolated from patients of accident injuries were used as the control.The chondrocytes were treated with HA at different doses.HSP70 expression in chondrocytes at both mRNA and protein levels was tested by PCR and Western blot analysis.Compared with control,both mRNA and protein levels of HSP70 were higher in chondrocytes from KBD and OA.However,HA at the dose of 500μg/mL significantly inhibited HSP70 expression levels in both KBD and OA groups(P<0.05).In conclusion,HSP70 is highly expressed in chondrocytes of patients of OA and KBD.HA intervention inhibits the upregulation of HSP70 in chondrocytes of OA and KBD patients and could be a promising agent for treatment of OA and KBD.

EFFECT OF SELENIUM ON IMMUNE FUNCTION OF ERYTHROCYTE IN KASHIN-BECK DISEASE

Objective To investigate the relationship between erythrocyte immune function and selenium (Se) level. Methods Forty-nine Kashin-Beck patients in endemic area aged 13-16 years were divided into two groups and were orally given either selenized yeast or sodium selenite to provide 200 μ g selenium per day for 12 weeks. Erythrocyte selenium level, glutathione peroxidase activity, the rosette formation rates of red blood cells complement receptor typeⅠ(CR1), the immune function of red blood cells, and circulating immune complexes(CIC) were determined. Results After supplementing with selenium for 12 weeks, erythrocyte selenium level, glutathione peroxidase activity, the rosette formation rates of red blood cells CR1 were significantly increased. But the difference in rosette formation rates of IC and CIC content was not significant between before and after Se supplementation. Conclusion The increase of the immune function of the erythrocyte by selenium-supplement may be one of the effective mechanisms for the prevention of Kashin-Beck disease.

Cold weather and Kashin-Beck disease

Kashin-Beck disease(KBD)is an endemic osteoarthropathy.Its distribution region covers a long and narrow belt on the Pacific side and belongs to continental climate with short summer,long frost period,and large temperature differences between day and night.In particular,KBD patients are typically scattered in the rural areas with seasonal features such as cold winters and rainy autumns.Etiological studies have demonstrated that the carrier of pathogenic factors is the grains produced in endemic areas.Risk factors for KBD include fungal contamination of grains due to poor storage conditions associated with cold weather.The epidemiological characteristics of KBD include agricultural area,early age of onset,gender equality,family aggregation,regional differences,and annual fluctuations.A series of preventive measures have been successfully taken in the past decades.National surveillance data indicate that the annual incidence of KBD is gradually declining.

STUDIES ON BLOOD GLUTATHIONE PEROXIDASE PROTEIN LEVEL OF CHILDREN IN KESHAN DISEASE AND KASHIN-BECK DISEASE AREAS

Objective To oberve the change in blood glutathione peroxidase (GSH-Px) protein levels of residents in the low-selenium (Se) area by contrasting the blood GSH-Px protein level of the children in the Keshan disease area with those in the Kashin-Beck disease and non-endemic areas. Methods GSH-Px protein levels were measured by enzyme-linked immunosorbent assays (ELISA). The Se content and GSH-Px activity were assayed by the 2,3-diaminonaphthalene spectrofluorimetric method and glutathione reductase-coupled method respectively. Results ①The blood Se content and GSH-Px protein level of children in Keshan disease area (Moding) were significantly lower than those in Xi’an non-endemic area, however, there was no significant difference when compared with the low-Se non-endemic area; ②The blood Se content, GSH-Px activity and GSH-Px protein level of children in the Kashin-Beck disease area (Yulin) were significantly lower than those of children in two non-endemic areas and in the Keshan disease area; ③The blood Se content and GSH-Px activity were positively correlated to the GSH-Px protein level respectively. Conclusion These results indicate that the blood GSH-Px protein level is decreased in the low-Se residents. The Se status not only affects the GSH-Px activity but also regulate the GSH-Px protein level.

X-ray image characteristics and related measurements in the ankles of 118 adult patients with Kashin-Beck disease

Background Kashin-Beck disease （KBD） is a chronic,degenerative osteoarthropathy that causes severe skeletal deformation.Although many researchers have proven that almost all KBD patients who showed an increaseing proximo-distal gradient had radiographic abnormalities of the ankle,few detailed description of radiographic changes in the ankles of patients with KBD has been reported,especially for variable measurements of ankle changes.The purpose of this study was to demonstrate the radiographic characteristics of the ankles of adult KBD patients.Methods One hundred and eighteen adult KBD patients from september to October 2010 in Rongtang county in China were examined with lateral radiographs of the right ankle.The morphological abnormalities in the talus,calcaneus,navicular bone,distal tibia,and joint space were analyzed,and the calcaneus length,height,length-height ratio,tuber angle,front angle,plantar angle,and distal tibia anteroposterior （AP） length were measured using Riepert＇s method.Results Eighty-one patients （68.6％） had abnormal ankle radiographs; 72 （88.9％） patients had talus changes,69 （85.2％） patients had calcaneus changes,28 （34.6％） patients had navicular bone changes,and 48 （59.2％） patients had distal tibia changes.For 118 KBD patients,the average calcaneus length was 7.4 cm,height was 4.3 cm,and the length-height ratio was 1.7.The calcaneus tuber angle was 28.2°,front angle was 38.0° and the plantar angle was 74.2°.The distal tibia anteroposterior length was 4.05 cm.Compared with 50 normal adults （control group）,significant differences were found for the calcaneus length,the calcaneus length-height ratio,and the distal tibia AP length.Conclusions Patients with KBD have characteristic abnormalities on ankle radiographs; talus depression and deformity,calcaneus shortening deformity,and distal tibia deformity with AP length widening were the most typical changes.

Soil selenium concentration and Kashin-Beck disease prevalence in Tibet,China

In order to investigate the correlation between the prevalence of Kashin-Back disease(KBD)and Se concentrations,natural soil samples and cultivated soil samples were collected from southeastern Tibet,China;and the soil Se concentrations were measured by atomic fluorescent spectrophotometer.It was found that the mean Se concentrations of natural soil samples in KBD areas,from the first layer to the third layer,were 0.17 mg/kg,0.11 mg/kg,and 0.10 mg/kg,respectively,and in nondisease areas were 0.21 mg/kg,0.24 mg/kg,and 0.13mg/kg,respectively.The mean Se concentrations of cultivated soil samples were 0.10 mg/kg in KBD areas and 0.23 mg/kg in non-disease areas,respectively.Soil Se concentrations in KBD areas were lower than that in non-disease areas,and the mean concentrations of soil Se in Tibet were lower than the average of China(0.29 mg/kg).Therefore,there is a close relationship between soil Se concentrations and KBD in Tibet.More studies should be concentrated on the impacts of Se deficiency in soils and its relationship with Se concentrations in food-grain and the human body in Tibet areas.

Relationship between selenium concentration in child hair and the distribution of Kashin-Beck Disease in Tibet, China

The focus of this study is to determine the relationship between selenium(Se)concentrations in child hair and the distribution of Kashin-Beck disease(KBD)in Tibet.The hair of children in typical KBD-affected areas and non-disease areas was collected,and its Se concentrations were measured by atomic fluorescent spectrometry.The Se concentrations in child hair were(0.13±0.04)and(0.18±0.07)μg/g respectively in KBD areas and non-disease areas respectively,which are significantly different.In areas supplemented with Se in Tibet,the Se concentration of hair in KBD-affected areas was higher than that in non-diseased areas,being(0.28±0.03)and(0.18±0.04)μg/g respectively.There is a close relationship between the Se concentrations in hair and KBD distribution.Se supplement increase the Se concentrations in the human body.Appropriate Se supplementation is important to prevent KBD in Tibet.

Effects of moniliformin and selenium on human articular cartilage metabolism and their potential relationships to the pathogenesis of Kashin-Beck disease

Objective： To investigate the effects of mycotoxin moniliformin （MON） on the metabolism of aggrecan and type 11 collagen in human chondrocytes in vitro and the relationship between MON and Kashin-Beck disease （KBD）. Methods： Human chondrocytes were isolated and cultured on bone matrix gelatin to form an artificial cartilage model in vitro with or without MON toxin. Cell viability was determined by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assay. The expression of aggrecan and type II collagen in the cartilage was determined using immunocytochemical staining. Results： MON toxin inhibited chondrocyte viability in dose-dependent and time-dependent manners. MON reduced aggrecan and type Ⅱ collagen syntheses in the tissue-engineered cartilage. MON also increased the expression of matrix metalloproteinase-1 （MMP-1）, MMP-13, BC4 epitopes, and CD44 in cartilages. However, the expression of 3B3（-） epitopes in cartilages was inhibited by MON. Selenium partially alleviated the damage of aggrecan induced by MON toxin. Conclusion： MON toxin promoted the catabolism of aggrecan and type II collagen in human chondrocytes.

Spatial Analysis and Biogeochemical Cycles: A Comparative Study of Kashin-Beck Disease Villages and Non-disease Villages in Linzhou County, Tibet

This study investigated Linzhou County in Tibet, which currently hosts the most serious outbreak of Kashin-Beck disease(KBD) in China. This study uses the geographical detector(GeoDetector) algorithm to measure the influences that several risk factors have on KBD prevalence and validates the spatial analysis results with environmental chemistry. Based on a comprehensive examination of 10 potentially related spatial factors and an environmental chemistry analysis of the soil-water-grain-human biogeochemical cycle in the local KBD and non-KBD villages, four main conclusions are drawn.(1) KBD in Linzhou County is a consequence of multiple interrelated environmental factors, of which the most important controlling factor is the stratum factor.(2) The concentrations of selenium(Se) in all environmental media(soil, water, and food) and human tissue in the KBD villages in Linzhou County are lower than those of the non-KBD villages.(3) The intake of Se and chromium(Cr) by local residents is seriously insufficient, especially the average daily dose by ingestion(ADD) for Se in the KBD village, which is only about 4% of the World Health Organization(WHO) recommended lower limit for adult elemental intake.(4) We speculate that the main cause for the local KBD outbreak is a lack of Se in the stratum. This absence leads to a serious Se deficiency in the local population through ecosystem migration and transformation, which will eventually lead to an endemic biogeochemical Se deficiency.

STUDIES ON LYMPHOCYTE SUBSETS, HUMORAL IMMUNITY AND THEIR RELATIONSHIP WITH SELENIUM IN PATIENTS WITH KASHIN-BECK DISEASE

Objective To study the humoral immunity status and distribution pattern of lymphocyte subgroups of peripheral blood mononuclear cell (PBMC) in patients with Kashin-Beck Disease (KBD), and their relationship with erythrocyte selenium. Methods 23 X-ray diagnosed patients, 22 age- and sex- matched healthy children in KBD affected area (KAA), and 25 in KBD non-affected area (KNAA) were randomly selected. Immunohistochemistry with monoclonal antibodies anti-CD4, anti-CD8, anti-CD20 was conducted to analyze the lymphocyte subsets. Serum IgM, IgA, IgG, Complement C3 and C4 were assayed using rate nephelometry (Array 360 System, USA). The contents of erythrocyte selenium was determined by 2,3-diaminonaphthalene fluorescence assay. Results CD4+ and CD8+ cells percentage in PBMCs and serum IgA were significantly lower in KAA than those in KNAA(P< 0.05). CD20+ percentage in KAA displayed a decreasing trend compared to KNAA, although not statistically significantly. No statistical differences were found in CD4/CD8 ratio, serum IgG, IgM, C3 and C4 levels. Erythrocyte selenium level in KAA still showed a pronounced decrease compared to that in KNAA. Correlation analysis showed that erythrocyte selenium contents had a strong association with the CD4 cell percentage (r= 0.625, P< 0.05), as well as serum IgA (r= 0.462, P< 0.05). In addition, a moderate correlation between the serum IgA and CD4+ percentage (r= 0.130, P> 0.05) was found. Conclusion These results suggested that children in KAA had a comparably low cellular immunity level and their humoral immunity status was also in a state of moderate immune suppression. Of this immune disorder in Kashin-Beck disease patients, selenium deficiency probably played a critical role via affecting the distribution pattern of peripheral blood lymphocyte. Selenium-deficiency and immune impairment maybe both have something to do with the cause-effect chain of KBD.

Application of Finite Element Analysis in Biomechanical Research of Degenerative Diseases of Lumbar Spine

As the elderly population continues to grow, the number of patients with low back pain is gradually increasing. Among them, Lumbar Degenerative Diseases (LDD) is one of the major contributors to low back pain. Biomechanical in vivo studies of the lumbar spine are mainly performed by implants or imaging data to record the real-time changes of form and stress on the intervertebral disc during motion. However, the current developments are slow due to the technological and ethical limitations. In vitro experiments include animal experiments and cadaver experiments, which are difficult to operate or differ greatly from normal human structures, and the results still need to be verified repeatedly to test their accuracy. As for finite element method, it is relatively low cost and can repeat the experimental results. Therefore, we believe that finite element analysis plays an extremely important role in biomechanical research, especially in analyzing the relationship between different surgical models and the degeneration caused by different mechanics.

Mechanopathology of red blood cell diseases—Why mechanics matters

During the onset of a disease a cell may experience alterations in both the composition and organization of its cellular and molecular structures.These alterations may eventually lead to changes in its geometrical and mechanical properties such as cell size and shape,deformability and adhesion.As such,knowing how diseased cells respond to mechanical forces can reveal ways by which they differ from healthy ones.Here,we will present biomechanistic insights into red blood cell related diseases that manifest mechanical property changes and how they directly contribute to the pathophysiology of diseases.By conducting cell and molecular mechanics studies,not only can we elucidate changes in the structure-property-function relationship of diseased cells,we can also exploit the new knowledge gained to develop biomechanics based devices that may better detect and diagnose these diseases as well as help identify important biomechanical targets for possible therapeutic interventions.