Hyaluronic acid and glucosamine sulfate for adult Kashin-Beck disease: a cluster-randomized, placebo-controlled study.

To evaluate the efficacy and safety of hyaluronic acid (HA) and glucosamine sulfate (GS) in alleviating symptoms and improving function of Kashin-Beck disease (KBD). A cluster-randomized, placebo-controlled trial was conducted in 150 patients with KBD. Participants were randomly allocated to receive intra-articular injection hyaluronic acid (IAHA) for 4 weeks, oral GS for 12 weeks, or oral placebo for 12 weeks. The primary outcome measures were 20 % and 50 % reductions in pain from baseline measured by the Western Ontario and McMaster Universities Osteoarthritis (WOMAC) index. Secondary outcome measures included WOMAC index parameters of pain, stiffness, and physical function. The third outcome measure was mean change in Lequence score. HA and GS were effective in reducing WOMAC pain by 20 % (differences of 43.5 % and 25.4 %) and 50 % (differences of 43.4 % and 26.9 %). Both HA and GS significantly reduced WOMAC pain, WOMAC stiffness, and WOMAC normalized score compared with placebo group (all P < 0.05). IAHA was significantly more effective than oral GS in improving WOMAC normalized score (P = 0.034), pain (P = 0.002), stiffness (P = 0.018), and function (P = 0.044). The results indicate that HA and GS were more effective than placebo in treating KBD and HA was more effective than GS.

[Transmission disequilibrium test for 15 short tandem repeat loci in Kashin-Beck disease and their interaction with low selenium].

OBJECTIVE: To identify the genetic susceptibility to Kashin-Beck disease (KBD) and explore the interaction between low selenium (Se) and the susceptibility gene loci in KBD. METHODS: The DNA samples collected from 23 KBD nuclear families were analyzed using PCR and GeneScan Analysis 3.7 and Genotyper3.7 software. The haplotype relative risk (HRR) and transmission disequilibrium test (TDT) were used to test the data of the genotypes. The serum selenium (Se) concentration was measured by atomic fluorescence spectrometry, and the interaction between low Se and the susceptibility loci was calculated using a binary logistic regression. RESULTS: In the 23 nuclear families, the alleles of D2S151 (248 bp), D2S305 (320 bp), and D11S4094 (194 bp) showed significant correlation to KBD (P<0.05). Serum Se concentrations in the studied individuals was 0.037 µg/ml. No significant statistical interaction was observed between low Se exposure and the susceptibility loci (P>0.05). CONCLUSION: The polymorphisms in the STR loci D2S305, D2S151, and D11S4094 or the polymorphism loci near them might been related to KBD susceptibility. Low Se exposure shows no significant interaction with the susceptibility loci.

[Association of genetic polymorphism of 14 STR loci on chromosome 2 with Kashin-Beck disease].

OBJECTIVE: To analyze the differences of genetic polymorphism of 14 STR loci on chromosome 2 between KBD patients and controls living in and outside of KBD catchment areas. METHODS: Blood samples anticoagulated with EDTA were collected from 135 unrelated individuals of Han population in Shaanxi Province, which included 45 samples from KBD patients, 45 from normal residents living in the KBD catchment areas, and 45 from normal residents outside of the KBD catchment areas. The DNA was extracted from the blood samples for PCR amplification of relevant fragments. The amplified products were analyzed using the ABI 3730 Genetic Analyzer. RESULTS: The allele numbers for 14 STR loci (D2S286, D2S165, D2S160, D2S2211, D2S367, D2S125, D2S206, D2S117, D2S142, D2S2333, D2S126, D2S325, D2S364, D2S337) in the KBD patients were 8, 11, 7, 7, 9, 10, 11, 11, 8, 10, 11, 10, 6 and 9, respectively. Different allele numbers for 14 STR loci were found in the normal residents in the KBD catchment areas (7, 10, 6, 7, 7, 8, 10, 10, 8, 8, 11, 8, 7 and 7) and those outside of the KBD catchment areas (8, 11, 7, 7, 10, 9, 11, 11, 7, 9, 11, 7, 8 and 7). There were significant differences in the allele frequencies in the D2S165 and D2S2333 locus between the KBD patients and the normal residents living in and outside of the KBD catchment areas (P > 0.05). Significant difference in the allele frequencies in the D2S160 loci was found between the KBD patients and the normal residents living in the KBD catchment areas (P = 0.046). Significant difference in the allele frequencies in the D2S364 loci was found among the three groups of participants (P = 0.046). CONCLUSION: The allele distribution patterns of the D2S165 and D2S2333 locus in KBD patients are different from normal people.

[Effects of selenium and/or iodine deficiency on chondrocyte apoptosis in rats].

OBJECTIVE: To explore the effects of selenium and/or iodine deficiency on chondrocyte apoptosis in articular cartilage in rats. METHODS: Forty-eight Sprague-Dawley rats were randomly divided into selenium deficiency group, iodine deficiency group, combined selenium and iodine deficiency group, and control group. Chondrocyte apoptosis was detected by terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) method, and Bcl-2 and Bax in articular cartilage were stained by immunohistochemistry in F3 generation of rats. RESULTS: In articular cartilage, the positive rate of apoptotic chondrocytes stained by TUNEL in the upper and middle zones in selenium deficiency group, iodine deficiency group, and combined selenium and iodine deficiency group (all P < 0.05) were significantly higher than that in control group. The apoptotic chondrocytes were prominent in the middle zone. The positive percentage of chondrocytes apoptosis was not significantly different among these three groups (P > 0.05). Compared with the control group, the expressions of both Bcl-2 and Bax were significantly higher in the upper and middle zone in the selenium deficiency group, iodine deficiency group, and combined selenium and iodine deficiency group (all P < 0.05); however, the expressions of Bcl-2 and Bax were not significantly different among these three groups (P > 0.05). CONCLUSION: Selenium and/or iodine deficiency may induce chondrocyte apoptosis.

Features of intervertebral disc degeneration in rat's aging process.

OBJECTIVE: The age-related change is important part of degenerative disc disease. However, no appropriate animal model or objective evaluation index is available. This study aimed to investigate the features of intervertebral disc degeneration in aging process of rats. METHODS: 22-month-old Sprague-Dawley (SD) rats were used as spontaneously occurring intervertebral disc degeneration models and 6-month-old rats as young controls. Expression of collagen types II and X was measured by immunohistochemistry. Degenerations of intervertebral discs were scored according to Miyamoto's method. Numbers and areas of afferent vascular buds were measured. The thicknesses of non-calcified and calcified layers were measured and statistically analyzed. RESULTS: There were less collagen type II expression and more collagen type X expression in the calcified layer of the cartilage endplates and nucleus pulposus in the rats of the aged group than in the young control. There were fewer and smaller afferent vascular buds in the rats of the aged group than in the young control group. The ratio of the non-calcified to the calcified layers in the rats of the aged group significantly decreased, compared with that of the young control group (P<0.01). CONCLUSION: Rats can spontaneously establish intervertebral disc age-related degeneration. The expression of collagen types II and X, numbers and areas of afferent vascular buds, the ratio of the non-calcified to the calcified layers, and water and glycosaminoglycan contents in the nucleus pulposus are sensitive indexes of intervertebral disc degeneration.

[Genetic polymorphism of 15 STR on chromosome 2 and 11 in Shaanxi Han people in China].

OBJECTIVE: To investigate the genetic polymorphism of 15 short tandem repeat(STR)loci on chromosome 2 and chromosome 11 in Shaanxi Han people in China. METHODS: Fluorescence-based gene scan technique was used to examine the genetic polymorphism of 15 STR loci in 175 unrelated individuals from Chinese Han population in Shannxi province. RESULTS: The number of alleles D2S335, D2S396, D2S338, D2S2382, D2S305, D2S151, D2S2368, D2S391,D11S912, D11S4090, D11S4147, D11S4190, D11S4149, D11S4126, and D11S4094 was 11,11,11,10,8,8,9,12 ,7,11,8,10,5,5, and 6. The distribution of allele frequencies of the 15 STR was consistent with Hard-Weinberg equilibrium (P > 0.05). Heterozygosity (H) value was 0.4216 to approximately 0.8517, the average power of discrimination (DP) was 0.6568 to approximately 0.9598, polymorphism information content (PIC) was 0.4078 to approximately 0.8366, and probability of paternity exclusion (EPP) was 0.3135 to approximately 0.8537. CONCLUSION: The 15 STR loci have relatively high genetic polymorphism in Shaanxi Han population, which provides the genetic structure of Chinese Han groups, and is also useful in anthropology and forensic science.

[Familial aggregation and sibling heritability in Kashin-Beck disease].

OBJECTIVE: To explore the family aggregation and the role of hereditary factors in the pathogenesis of Kashin-Beck disease (KBD). METHODS: With a stratified sampling method, the general population of 14 villages of Linyou County were studied, from whom 225 KBD probands were selected using systematic sampling at the rate of (1/2). A total of 304 siblings of the probands were ascertained, and in these sibling pairs, the segregation ratio, heritability in different age groups and weighted mean heritability of the siblings were estimated using the methods of Li-Mantel-Grart and Falconer. RESULTS: The KBD distribution scope in the KBD families exceeded the scope of binomial distribution (P<0.001), suggesting obvious family aggregation. The prevalence rate in the siblings of the KBD pedigree was 19.41% (59/304), significantly higher than that in the 14 KBD villages [10.90% (1180/10823), chi2=21.62, P<0.001]. The segregation ratio and heritability in the siblings of the KBD pedigrees were 0.061 and 28.61%, respectively. CONCLUSION: As a polygenetic inheritance disease, KBD exhibits obvious familial aggregation, and genetic susceptibility accounts for (1/4) of the risk factors for KBD.

[Analysis of genetic polymorphisms of 10 STR loci on chromosome 11 in a Shaanxi Han population].

D11S1760, D11S4102, D11S4116, D11S4207, D11S4162, D11S914, D11S4127, D11S917, D11S4146 and D11S915 of 10 short tandem repeat (STR) loci on chromosome 11 were analyzed by fluorescence-based gene scan technique to understand the genetic polymorphisms of those STR loci on chromosome 11 in a Han population in Shaanxi province. The results showed that the number of alleles and genotypes observed at loci D11S1760, D11S4102, D11S4116, D11S4207, D11S4162, D11S914, D11S4127, D11S917, D11S4146 and D11S915 were 17, 11, 15, 11, 4, 6, 7, 12, 11 and 13 for alleles and 41, 18, 36, 30, 8, 10, 16, 30, 29 and 32 for genotypes, respectively. All the 10 loci were in Hardy-Weinberg equilibrium. The heterozygosities of each STR locus was 86.72%, 67.95%, 83.90%, 85.96%, 58.18%, 57.63%, 72.60%, 72.73%, 77.87% and 86.40%. It concluded the 10 loci on chromosome 11 were relatively highly genetic polymorphic in Han populations and could provide useful markers for genetic studies.

[Genetic polymorphism analysis of 8 short tandem repeat loci on chromosome 2 in Chinese Han population in Shaanxi Province].

OBJECTIVE: To analyze the genetic polymorphism of 8 short tandem repeat (STR) loci on human chromosome 2 in Chinese Han population in Shaanxi Province. METHODS: Blood samples anticoagulated with EDTA were collected from 176 unrelated Chinese Han individuals in Shaanxi Province. The DNA was extracted for PCR amplification of the relevant fragments, and the amplified products were analyzed using the ABI 3730 Genetic Analyzer. RESULTS: On human chromosome 2, the loci D2S112, D2S162, D2S2330, D2S2216, D2S347, D2S259, D2S319 and D2S168 had 7, 11, 9, 8, 9, 9, 8 and 13 alleles, respectively, with 15, 33, 23, 18, 13, 12, 25 and 33 genotypes for the corresponding alleles. The genotype distribution of all the 8 loci met Hardy-Weinberg equilibrium. The heterozygosities for the 8 STR loci were 0.6985, 0.8274, 0.8042, 0.6816, 0.6541, 0.5213, 0.8432 and 0.8091, with polymorphic information content of 0.6911, 0.8199, 0.7891, 0.6809, 0.6388, 0.5187, 0.8372 and 0.8049, respectively. CONCLUSION: The 8 loci on chromosome 2 have high heterozygosity and polymorphic information content in Chinese Han population, suggesting their value as useful genetic markers.

[Butenolide induces apoptosis of cultured chondrocytes: study of its mechanism].

OBJECTIVE: To observe cell apoptosis and Bcl-2 and Bax expression changes of chondrocytes induced by butenolide (BUT) and the inhibitory effect of selenium against BUT-induced chondrcyte apoptosis, to gain insights into the mechanism by which BUT induces chondrcyte apoptosis. METHODS: Cartilage tissue reestablished from human fetal articular chondrocytes in vitro were treated with BUT at the concentrations of 0.1, 1.0 and 5.0 microg/ml and with the protective factor selenium. TUNEL method was used to detect chondrocyte apoptosis, which was quantified by flow cytometry. Immunohitochemistry was performed to analyze the expression of Bcl-2 and Bax in the reestablished cartilage tissue. RESULTS: BUT exposure induced chondrocyte apoptosis, and the apoptosis rate increased with the concentration increment of BUT from 0 to 1.0 mg/ml, resulting also increased positive expression rate of Bcl-2 and Bax(P<0.05). The apoptosis rate of chondrocytes in BUT+ selenium group was significantly lower than that of BUT groups (P<0.05), as was the positivity rate of Bcl-2 and Bax expression (P<0.05). CONCLUSION: BUT induces chondrocyte apoptosis in positive relation with BUT concentration (from 0 to 1.0 mg/ml) and causes increased expressions of Bcl-2 and Bax. Selenium can inhibit the chondrocyte apoptosis induced by BUT.

[Mechanism of chondrocyte apoptosis in Kashin-Beck disease and primary osteoarthritis: a comparative study].

OBJECTIVE: To investigate chondrocyte apoptosis and the expressions of Bcl-2, Bax, Fas and iNOS in the articular cartilage between Kashin-Beck disease (KBD) and primary osteoarthritis (OA) and explore the difference in pathogenesis between the two diseases. METHODS: The articular cartilage specimens were collected from 15 normal human subjects, 15 adult patients with KBD and 15 with OA. Chondrocyte apoptosis was detected by TUNEL method, and the expressions of Bcl-2, Bax, Fas and iNOS in articular cartilage were examined with B-SA immunohistochemistry. RESULTS: The percentages of apoptotic chondrocytes positive for TUNEL staining in the articular cartilage were significantly higher in patients with KBD and OA than in normal control subjects (F=20.90-53.16, df=42, P<0.01), and the erosive areas of the articular cartilage contained greater percentage of apoptotic chondrocytes than the non-erosive areas in the same patient with KBD (t=4.154, df=28, P<0.01) or OA (t=6.004, df=28, P<0.01). No significant difference was noted in the positive apoptotic chondrocytes between KBD and OA (t=1.329-1.362, df=28, P>0.05). The percentage of chondrocytes positive for Bcl-2, Bax, Fas and iNOS were significantly higher in KBD and OA patients than in the control subjects (F=25.46-215.31, df=42, P<0.01), and significant differences were observed in Bcl-2, Bax, Fas and iNOS expressions between the erosed areas and non-erosed areas in articular cartilage in patients with KBD (t=2.608-7.77, df=28, P<0.05) and OA (t=2.278-5.413, df=28, P<0.05), but their expressions showed no significant difference between the two diseases (t=0.284-1.590, df=28, P>0.05). CONCLUSION: There was no significant difference in apoptotic chondrocytes and Bcl-2, Bax, Fas and iNOS expressions in the cartilage between adult patients with KBD and OA.

[Comparison of apoptosis of articular chondrocytes in the pathogenesis of Kashin-beck disease and primary osteoarthritis].

OBJECTIVE: To investigate chondrocyte apoptosis and expression of Fas and inducible nitric oxide synthase (iNOS) in articular cartilage in the pathogenesis of Kashin-beck disease (KBD) and primary osteoarthritis (OA). METHODS: The collected samples of articular cartilage were divided into three groups: normal control (15 cases), KBD adults (15 cases) and OA (15 cases). Chondrocyte apoptosis was detected by terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling method, and Fas and iNOS in articular cartilage were stained by immunohistochemistry. RESULTS: The positive percentages of chondrocyte apoptosis stained in articular cartilage of KBD and OA were significantly higher than that of the control (P < 0.01), and the positive percentage of chondrocytes apoptosis in the eroded areas of articular cartilage were significantly higher than in the non-eroded areas in articular cartilage of the same patient with KBD and OA (P < 0.05). There was no significant difference in positive percentage of chondrocytes apoptosis between KBD and OA. The positive percentages of Fas and iNOS in chondrocytes were significantly higher in KBD and OA than in control (P < 0.01). Significant differences in Fas and iNOS expression between the eroded areas and non-eroded areas were seen in articular cartilage of patients with KBD and OA (P < 0.05), but such difference did not exist between KBD and OA. CONCLUSION: Cell apoptosis seems to be associated with the pathogenesis of both KBD and OA. Fas and iNOS might mediate chondrocyte apoptosis.