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The Zhuanggu Guanjie herbal formula has been a famous Chinese prescription for treating bone diseases since time immemorial. The anti-osteoarthritis (OA) properties of this botanical prescription are well documented in the Chinese Pharmacopoeia. However, the detailed mechanisms behind the phenomenon have not been elucidated. Hence, we aimed to investigate the anti-OA efficacy of the Zhuanggu Guanjie herbal formula and its underlying mechanism. The anti-OA properties of Zhuanggu Guanjie capsule (ZGC) were determined by the cytokine contents and inflammatory-related proteins, which were measured by RT-PCR, flow cytometry, Western blot, and laser confocal assay in ATDC5 cells. The levels of interleukin-6, tumor necrosis factor-α, inducible nitric oxide synthase, cyclooxygenase-2, and prostaglandin synthesis E2 have been markedly reduced after being treated with ZGC for 48 h in a dose-dependent manner. Furthermore, ZGC prevented the translocation of NF-κB from the cytosol to the nucleus. On the other hand, we used the mono-iodoacetate (MIA)-induced OA model to confirm the in vivo efficacies of this herbal formula. Oral administration of ZGC attenuated MIA-induced OA damage through changes in histopathological and knee joint volumes. The serum matrix metalloproteinase-13 contents in the ZGC treatment group declined as compared to those in the MIA model group. Through our in vitro and in vivo studies, we confirmed the anti-OA efficacy of ZGC and uncovered its detailed mechanism, and this treatment shed light on OA pathophysiology.

Toll‐like receptors (TLRs) are an important family of pattern‐recognition receptors that recognize a series of pathogen‐associated molecular patterns to trigger the host immune response. In the present study, we identified three TLR13 genes (i.e., LcLTR13a, LcTLR13b, and LcTLR13c) in American bullfrogs, Lithobates catesbeianus, and explored their constitutive and pathogen‐induced expression profile in different tissues. The open reading frames of LcLTR13a, LcTLR13b and LcTLR13c are 2811, 2844, and 2814 bp long, encoding 955, 947, and 933 amino acids, respectively. Phylogenetic analysis showed that three LcTLR13 genes grouping in different subclades indicate the gene duplication of bullfrog TLR13 as in teleost. The three LcTLR13 genes were expressed in all the tissues examined, with the liver and spleen expressing the higher levels. Moreover, the expression levels of LcLTR13a, LcTLR13b, and LcTLR13c were significantly induced in the liver and spleen after bullfrogs were challenged with bacterial (i.e., Citrobacter freundii, Aeromonas hydrophila, and Edwardsiella tarda) pathogens. These results indicate that LcTLR13 genes play an essential role in the bullfrog antibacterial innate immune response, providing new insights into immune mechanisms of bullfrogs. [ABSTRACT FROM AUTHOR]

In recent years, bullfrog (Lithobates catesbeianus) has become one of the most commercially important amphibians for aquaculture in China, and its annual production has grown rapidly. However, bacterial diseases caused by Citrobacter freundii infection have resulted in enormous economic losses. The present study used RNA sequencing technology to analyze the transcriptomic profile and differentially expressed genes (DEGs) in the bullfrog liver and spleen post C. freundii infection. De novo assembly resulted in the generation of 77,556 L. catesbeianus transcripts, and 49,421 of these genes were successfully annotated to the databases. Overall, 1487 DEGs were identified in the liver group, including 788 upregulated and 709 downregulated DEGs. A total of 799 DEGs, including 311 upregulated and 488 downregulated genes, were detected in the spleen groups. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses revealed significantly enriched pathways, including pathways involved in metabolism and immune responses. The processes in which the DEGs were enriched were those related to metabolic activities, including the lipid metabolism pathway, amino acid pathway, immune‐related pathway, and inflammatory responses. The results of this study provide insight into the response of bullfrogs against C. freundii at the transcriptome level and reveal the mechanism of the immune response against C. freundii. [ABSTRACT FROM AUTHOR]

Background: Although glucose‐6‐phosphate dehydrogenase (G6PD) deficiency is the most common inherited disorder in the Chinese population, there is scarce evidence regarding the epidemiology, evolutionary origin, and malaria‐induced positive selection effects of G6PD‐deficient alleles in various Chinese ethnic populations. Methods: We performed a large population‐based screening (n = 15,690) to examine the impact of selection on human nucleotide diversity and to infer the evolutionary history of the most common deficiency alleles in Chinese populations. Results: The frequencies of G6PD deficiency ranged from 0% to 11.6% in 12 Chinese ethnic populations. A frequency map based on geographic information showed that G6PD deficiency was highly correlated with historical malaria prevalence in China and was affected by altitude and latitude. The five most frequently occurring G6PD gene variants were NM_001042351.3:c.1376G>T, NM_001042351.3:c.1388G>A, NM_001042351.3:c.95A>G, NM_001042351.3:c.1311T>C, and NM_001042351.3:c.1024C>T, which were distributed with ethnic features. A pathogenic but rarely reported variant site (NM_001042351.3:c.448G>A) was identified in this study. Bioinformatic analysis revealed a strong and recent positive selection targeting the NM_001042351.3:c.1376G>T allele that originated in the past 3125 to 3750 years and another selection targeting the NM_001042351.3:c.1388G>A allele that originated in the past 5000 to 6000 years. Additionally, both alleles originated from a single ancestor. Conclusion: These results indicate that malaria has had a major impact on the Chinese genome since the introduction of rice agriculture. [ABSTRACT FROM AUTHOR]

• Development of a Rapid and Efficient Detection System: We have developed an efficient RPA- Pf Ago detection system for the MTHFR C677T polymorphism, streamlining the genotyping process. • RPA Reaction, gDNA, and RPA- Pf Ago Optimization: Through meticulous optimization, we enhanced the efficiency and sensitivity of the MTHFR C677T detection system. • Clinical Validation: Our method has been clinically validated on samples and showed 100% concordance with Sanger sequencing, confirming the precision of the method. • Potential for Rapid, On-Site Genotyping: The RPA- Pf Ago system we have developed offers a tool for rapid on-site genotyping. This study introduces an efficient RPA- Pf Ago detection system for the MTHFR C677T polymorphism, proposing a potential strategy to simplify the genotyping process. By optimizing recombinase polymerase amplification (RPA) with Pyrococcus furiosus Argonaute (Pf Ago) nucleases, we achieved DNA amplification at a constant temperature. The assay was fine-tuned through meticulous primer and guide DNA selection, with optimal conditions established at 2.0 µL of MgAc, a reaction temperature of 42 °C, and a 10-minute reaction time for RPA. Further optimization of the Pf Ago cleavage assay revealed the ideal concentrations of MnCl 2 , guide DNA, molecular beacon probes, the Pf Ago enzyme, and the RPA product to maximize sensitivity and specificity. Clinical validation of 20 samples showed 100% concordance with Sanger sequencing, confirming the method's precision. The RPA- Pf Ago system is a promising tool for on-site genotyping, with broad applications in personalized medicine and disease prevention. [ABSTRACT FROM AUTHOR]