Objective We evaluated the association between monocyte chemotactic protein-1 (MCP-1) and osteoarthritis. Methods We searched PubMed, Cochrane Library, Embase, Web of Science, China National Knowledge Infrastructure (CNKI), VIP (Chinese database), and Wan Fang (Chinese database) (before May 10, 2020), with no language limitations. STATA version 12.0 and Revman version 5.3 were used for data analysis. The standard mean difference (SMD) and corresponding 95% confidence intervals (95% CIs) were calculated. Nine clinical studies, including 376 patients with osteoarthritis and 306 healthy controls, were evaluated. Results The combined SMDs of MCP-1 expression levels suggested that MCP-1 expression was significantly higher in patients with osteoarthritis than healthy controls (SMD = 1.97, 95% CI = 0.66–3.28, p = 0.003). Moreover, subgroup analysis implied that osteoarthritis patients from both Asians and mixed populations had higher MCP-1 expression levels than controls, whereas Caucasians did not (p > 0.05). Serum MCP-1 levels (SMD = 2.83, 95% CI = 1.07–4.6, p < 0.00001) were significantly higher in patients with osteoarthritis than in controls; however, this difference was not significant in synovial fluid and cartilage tissue. Subgroup analysis for ethnicity showed that MCP-1 levels were significantly higher in Chinese, Dutch, and Brazilian patients with osteoarthritis than in control groups, although significant differences were not observed for American and Italian subgroups. Conclusions Our meta-analysis demonstrated that MCP-1 expression levels were higher in patients with osteoarthritis than in healthy controls and that MCP-1 may play important roles in the progression of osteoarthritis. Serum MCP-1 levels may serve as a potential biomarker for the diagnosis of osteoarthritis.

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Sox8 genes, as members of the Sox family, have been studied widely in mammals. However, regulation of sox8 genes in teleosts has rarely been studied, and functional analysis of these genes in teleosts has rarely been performed. Here, two duplicates of sox8 genes were identified in Japanese flounder, Posox8a and Posox8b. The analysis of expression showed that Posox8a and Posox8b were expressed in Sertoli cells of the testis, indicating that they play important roles in development and functional maintenance of the testis. Positive selection and phylogenetic analysis found that both Posox8a and Posox8b underwent the purification selection during evolutionary and that sox8 was most likely to be the ancestor sox8a. These results suggested that both Posox8a and Posox8b had important biological functions after generation from three rounds of whole-genome duplication in Japanese flounder. The functional differentiation of Posox8a and Posox8b was verified using cell transfection and dual-luciferase reporter assays; Posox8a overexpression-promoted 3β-hydroxysteroid dehydrogenase expression and Posox8b overexpression-promoted cytochrome P450 aromatase (cyp19a1; P450arom) expression. Finally, combined with Posox8a and Posox8b expression analysis from 30 to 100 days after hatch, we speculated that Posox8a and Posox8b might participate in the process of sex differentiation and gonadogenesis by regulating sex hormone biosynthesis in the Japanese flounder. Our study is the first to demonstrate the possible mechanism of Posox8a and Posox8b in Japanese flounder sex differentiation and gonadogenesis, laying a solid foundation for functional studies of sox8 genes in teleosts. K E Y W O R D S dimorphic expression, evolutionary analysis, Japanese flounder, Posox8a and Posox8b, sex differentiation

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The P450 side-chain cleavage enzymes P450scc (Cyp11a) and 11β-hydroxylase (Cyp11b) play important roles in sex steroid and cortisol production. Here, two duplicates of cyp11 genes were identified in Japanese flounder (Paralichthys olivaceus): Pocyp11a and Pocyp11b, respectively. Phylogenetic analysis and amino acid sequence alignment revealed that Pocyp11a and Pocyp11b shared significant identity with sequences of other teleost fish species. The quantitative real-time polymerase chain reaction (qRT-PCR) results indicated that among the studied tissues, brain tissue showed the highest expression of Pocyp11a, followed by kidney and testis tissues, whereas Pocyp11b expression was highest in the testis. The expression patterns of these two genes showed sexual dimorphism, with both genes showing higher expression in the testis than in the ovary. In-situ hybridization analysis demonstrated that Pocyp11a and Pocyp11b mRNA were both detected in oocytes, spermatocytes, and Sertoli cells, indicating that they might be involved in hormone synthesis. The expression levels of Pocyp11a and Pocyp11b were significantly downregulated by treatment with 17α-methyltestosterone (17α-MT) in the testis and ovary in both in vivo and studies. In vivo studies showed that Pocyp11a and Pocyp11b transcripts were suppressed by 17β-estradiol (E 2 ) treatment in both the testis and ovary. In addition, in vitro studies showed that the expression level of Pocyp11b was decreased by treatment with E 2 , whereas that of Pocyp11a was largely unaffected.Moreover, the expression levels of Pocyp11a and Pocyp11b in the testis cell line were significantly upregulated after NR0b1 and NR5a2 (p < .05) treatment. These results indicate that Pocyp11a and Pocyp11b might play important roles in sex hormone biosynthesis. Our research can assist future studies of the mechanisms of steroid biosynthesis and functional differences between cyp11a and cyp11b in Japanese flounder. K E Y W O R D SJapanese flounder, sex hormones, NR5a2/NR0b1, Pocyp11a and Pocyp11b