Objective:To observe and compare the efficacy and safety of glucosamine sulfate (GS) and chondroitin sulfate (CS) in the treatment of adult Kashin-Beck disease (KBD), so as to provide effective medical evidence for the standardized treatment of adult KBD.Methods:A clinical randomized controlled trial was conducted in Fuyu County and Shangzhi City, KBD historical seriously ill areas in Heilongjiang Province. A total of 247 patients were selected according to the standard of "Diagnosis of Kashin-Beck Disease" (WS/T 207-2010). According to gender, age and KBD condition, they were randomly divided into GS and CS groups, 124 and 123 respectively. Follow up once a month to investigate the medication and clinical symptoms of patients, and distribute drugs for the next stage. Fasting blood and urine samples were collected before, during and at the end of treatment (0, 90 and 180 d). Serum interleukin (IL)-1β content and urine pyridine (PYD) level were measured by enzyme-linked immunosorbent assay (ELISA). The visual analogue scale (VAS) score, affected joints, self-evaluation of curative effect and side effects were evaluated through the questionnaire, joint dysfunction and drug efficacy were evaluated according to the criteria of "Evaluation of Therapeutic Effect of Kashin-Beck Disease" (WS/T 79-2011).Results:Expression of cytokines related to cartilage metabolism: at 180 d of treatment, serum IL-1β contents and urinary PYD levels in GS and CS groups were lower than those at 0 d of treatment ( Z = - 2.461, - 2.160, - 5.075, - 5.471, P < 0.05). VAS score: at 90 and 180 d of treatment, the scores of knee pain, stiffness and function in GS and CS groups were lower than those at 0 d of treatment ( P < 0.05); and at 180 d of treatment, the scores of knee stiffness and function in GS group were lower than those in CS group ( P < 0.05). Evaluation of affected joints: at 90 and 180 d of treatment, the scores of joint pain, swelling and stiffness in GS and CS groups were lower than those at 0 d of treatment ( P < 0.05). Self-evaluation of curative effect: at 180 d of treatment, the self-evaluation of curative of CS group were better than that at 90 d of treatment (χ 2 = 9.376, P < 0.05). Evaluation of side effects: at 90 and 180 d of treatment, the side effects in GS and CS groups were mainly gastrointestinal symptoms. Joint dysfunction score: at 90 d of treatment, the sum of effective rate and markedly effective rate in GS group was higher than that in CS group (χ 2 = 4.042, P < 0.05), but there was no significant difference between the two groups at 180 d of treatment (χ 2 = 0.869, P > 0.05). Conclusion:GS and CS have certain therapeutic effects on adult KBD, which can improve symptoms and reduce serum IL-1β content and urinary PYD level, but GS takes effects quickly, and its effect on improving joint stiffness and function are better than CS.

Objective:To explore the effects of HT-2 toxin on expressions of silent information regulator of transcription 1 (SIRT1) and autophagy and apoptosis pathway related proteins in cultured chondrocytes in vitro. Methods:The third-generation chondrocytes of SD neonatal rats aged 1 to 2 days were cultured in vitro and identified by toluidine blue staining and type Ⅱ collagen immunofluorescence staining. CCK-8 method was used to detect the proliferation of chondrocytes. According to the cell survival rate, 2, 4 and 8 ng/ml HT-2 toxin were selected for subsequent experiments, and the exposure time was 48 h. At the same time, a negative control group and a solvent (absolute ethanol) control group were set up. Western blotting was used to detect the expressions of SIRT1 and autophagy and apoptosis pathway related proteins [microtubule-associated protein 1 light chain 3 (LC3)-Ⅱ, LC3-Ⅰ, p62, Beclin1, Caspase-3, B-cell lymphoma-2 (Bcl-2) and Bcl-2-Associated X protein (Bax)] in each group. Results:After staining, the cells were identified as chondrocytes with high purity. The expression levels of SIRT1 protein in 2, 4, 8 ng/ml HT-2 toxin groups (0.69 ± 0.18, 0.46 ± 0.13, 0.35 ± 0.19) were significantly lower than that in negative control group (1.00 ± 0.39, P < 0.05). In 2, 4 and 8 ng/ml HT-2 toxin groups, the ratios of autophagy pathway related proteins LC3-Ⅱ and LC3-Ⅰ expressions (LC3-Ⅱ/LC3-Ⅰ, 1.47 ± 0.15, 1.37 ± 0.13, 1.81 ± 0.34) were higher than that in negative control group (1.00 ± 0.21, P < 0.05), and the expression levels of p62 protein in 4, 8 ng/ml HT-2 toxin groups (0.70 ± 0.04, 0.57 ± 0.01) were lower than that in negative control group (1.00 ± 0.15, P < 0.05). In 2, 4, 8 ng/ml HT-2 toxin groups, the expression levels of apoptosis pathway related protein Bcl-2 (0.61 ± 0.06, 0.54 ± 0.16, 0.47 ± 0.06) were significantly lower than that in negative control group (1.00 ± 0.14, P < 0.05), and the ratio of Bax to Bcl-2 protein expressions in 8 ng/ml HT-2 toxin group (Bax/Bcl-2, 3.27 ± 0.18) was higher than that in negative control group (1.00 ± 0.27, P < 0.05). The expression level of SIRT1 protein was significantly negatively correlated with the expression level of autophagy pathway related protein LC3-Ⅱ ( r = - 0.819, P < 0.01), and was significantly positively correlated with the expression level of p62 protein( r = 0.772, P < 0.01), but not with the expression level of Beclin1 protein ( r = 0.399 , P > 0.05); there was no correlation between SIRT1 protein expression and apoptosis pathway related protein Caspase-3 and Bax expressions ( r = - 0.297、- 0.284, P > 0.05), but there was a significant positive correlation with Bcl-2 protein expression ( r = 0.755, P < 0.01). Conclusion:HT-2 toxin may increase the expression of autophagy pathway related protein LC3-Ⅱ/LC3-Ⅰ, decrease the expression of p62 protein, and increase the apoptosis pathway related protein Bax/Bcl-2 by inhibiting the expression of SIRT1 protein in chondrocytes, resulting in abnormal autophagy and apoptosis, and finally leads to the injury of chondrocytes.

Objective:To observe and compare the therapeutic effects of glucosamine sulfate (GS) and diacerein (DCN) on adult Kashin-Beck disease (KBD).Methods:A clinical randomized controlled trial was conducted in the historical severe KBD areas Fanrong Township, Fulu Town, Long'anqiao Town, Lianghe Town, Shaowen Township of Heilongjiang Province, and 240 patients were selected according to the criteria of "Diagnosis of Kashin-Beck Disease" (WS/T 207-2010), then divided into GS and DCN groups (gender, age, and KBD condition balanced) via the random number table method, with 120 patients in each group. Followed up once a month to investigate the patient's medication and clinical symptoms, and distributed drugs for the next stage. Fasting blood samples and urine samples were collected before, during, and at the end of treatment (0, 90, and 180 days). Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum interleukin (IL)-1β level and urine pyridinol (PYD) level. Visual analog scale (VAS) scores, evaluation of affected joints, self-evaluated efficacy, and evaluation of adverse reactions were carried out through questionnaires. Joint dysfunction scores and medications efficacy determination were performed according to the "Judgment of Kaschin-Beck Disease Treatment Effect" (WS/T 79-2011).Results:Expression of cytokines related to cartilage metabolism: after 180 days of treatment, serum IL-1β levels, urine PYD levels in GS group and urine PYD levels in DCN group were lower than those in the same group at 0 day of treatment ( Z = - 2.332, - 5.420, - 5.204, P < 0.05). VAS scores: after 90 days of treatment, the pain, stiffness scores of patients in GS group and the pain, stiffness, and function scores in DCN group were lower than those in the same group at 0 day of treatment ( Z = - 2.612, - 2.359, - 3.637, - 2.881, - 2.238, P < 0.05); after 180 days of treatment, the pain, stiffness and function scores of patients in GS and DCN groups were significantly lower than those of the same group at 0 day of treatment ( Z = - 6.738, - 9.530, - 7.781, - 5.428, - 3.761, - 3.587, P < 0.01). Evaluation of affected joints: after 90 and 180 days of treatment, except for pain of weather changes in DCN group, the scores of symptomatic joints in the two groups were lower than those at 0 day of treatment ( P < 0.05). Efficacy self-evaluation: after 180 days of treatment, the self-evaluated efficacy ratio of DCN group was higher than that of GS group and the same group after 90 days of treatment (χ 2 = 4.165, 4.022, P < 0.05). Evaluation of adverse reactions: after 90 and 180 days of treatment, the main adverse reactions of patients in GS and DCN groups were gastrointestinal symptoms. Joint dysfunction scores: after 90 days of treatment, the sum of the effective rate and the markedly effective rate of GS group was higher than that of DCN group (χ 2 = 4.993 , P < 0.05); while after the 180 days of treatment, there was no significant difference between the two groups (χ 2 = 0.417 , P > 0.05). Conclusions:Both GS and DCN have a certain therapeutic effect on adult KBD and can improve clinical symptoms. The GS takes effect quickly, and long-term use can protect cartilage from inflammatory factors to a certain extent.

Objective:To investigate the degree of limb dysfunction in adult patients with Kashin-Beck disease (KBD), and the correlation between clinical grade of KBD and physical disability classification.Methods:Based on the monitoring data, using typical survey methods, 10 natural villages were selected as survey sites in the historical critical area of KBD in Heilongjiang Province in 2015. Patients over 40 years old with KBD were investigated by questionnaire, joint range of motion(ROM) examination, and X-ray film were performed. The degree of physical disability of the surveyed patients was evaluated according to the national standard of "Classification and Grading of Disability of the Disabled" (GB/T 26341-2010). The correlation between clinical classification of KBD and limb disability classification was analyzed.Results:A total of 137 adult patients with KBD were investigated, the age was (57.4 ± 9.9) years old. Among them, 84 were males and 53 were females; 95 were grade Ⅰ, 30 were grade Ⅱ and 12 were grade Ⅲ. The most common joint pain of upper limb was interphalangeal joint(126 cases, 126/137), followed by elbow joint (116 cases, 116/137); the lower limbs were mainly ankle joint (118 cases, 118/137) and knee joint (107 cases, 107/137). There were significant differences of detection rates in elbow, knee, ankle, hip and wrist joints dysfunction among different age groups ( P < 0.05). The detection rate increased with age. There was no correlation between the clinical grade of KBD and the classification of physical disability ( rs = - 0.142, P > 0.05). KBD patients accounted for the highest proportion of tertiary disability (60 cases, 60/137). The physical disability of male patients was more serious than that of female patients (χ 2 = 22.610, P < 0.01). Conclusions:In adults with KBD, interphalangeal joint pain is the most common in the upper limbs, and the ankle and knee joints are the most common in the lower limbs. There is no correlation between clinical grade of KBD and the level of physical disability. The degree of physical disability in male patients is higher than that in female patients.

Background Kashin-Beck disease (KBD) is an endemic osteoarthropathy, and its pathogenesis is still not entirely clear. Pathologically, many KBD changes are similar to those of osteoarthritis (OA). Therefore, this study aimed to identify changes in the levels of potential urinary biomarkers for OA, including C-telopeptide of type II collagen (uCTX-II), type II collagen cleavage neoepitope (uC2C), pyridinoline (uPYD), and uHelix-II, among adults with KBD. Methods Urinary samples of 83 external control (EC) subjects, 91 KBD patients, and 86 internal control (IC) subjects were tested by ELISA after the subjects completed a questionnaire and X-ray examination. Results The medians of the four markers in the KBD group were higher than those in the EC group and those in the IC group. The medians in the grade II KBD group were higher than those in the grade I group but were not statistically significant (P = 0.301, P = 0.408, P = 0.204, and P = 0.898 for uCTX-II, uC2C, uPYD, and uHelix-II, respectively). The area under the curve (AUC) of uCTX-II (0.775) was higher than that of the others (0.672, 0.639, and 0.628 for uC2C, uPYD, and uHelix-II, respectively). Conclusion The levels of uCTX-II, uC2C, uPYD, and uHelix-II were elevated in adults with KBD and showed an increasing trend as the severity of KBD increased. The prediction accuracy of uCTX-II was more useful than that of the others for assisting in the diagnosis of KBD.

Atrial Fibrillation (AF) is common in the elderly and Sestrins (Sesns) have been suggested to prevent age-related pathologies. The aim of this study was to investigate the effects of Sesns in AF. Clinical data were collected and a small sample of atrial appendage and atrium was obtained from patients undergoing valve repairment. The expression of Sesn1, Sesn2, and Sesn3 was significantly higher in patients with permanent atrial fibrillation (PmAF) than that in sinus rhythm (SR), and further greater in the left atrium than the right in PmAF patients. Superoxide anion and malondialdehyde were enhanced and positively correlated to the protein expression of Sesn1/2/3. Reactive oxygen species (ROS) production and Ca2+ overload were significantly decreased and cell survival was enhanced by overexpression of Sesns 1/2/3 in cultured HL-1 cells. Conversely, knockdown of Sesn1/2/3 resulted in significantly increased ROS and Ca2+ overload. In addition, the overexpression of Sesn1/2 significantly reduced the proliferation of fibroblasts, as well as decreased the protein expression of collagen and fibronectin1 in angiotensin II-stimulated cardiac fibroblasts. Our study demonstrated for the first time that Sesns expression is significantly up-regulated in AF, which therefore may protect hearts against oxidative damage and atrial fibrosis.