ObjectiveTo investigate the effect and underlying mechanism of the Wulao Qisun prescription on pathological new bone formation in ankylosing spondylitis （AS）. MethodsSynovial fibroblasts were isolated from the hip joints of AS patients and observed under a microscope to assess cell morphology. The cells were identified using immunofluorescence staining. The isolated AS fibroblasts were divided into blank group， low drug-containing serum group， medium drug-containing serum group， high drug-containing serum group， and positive drug group. After drug intervention， cell proliferation was measured using the cell counting kit-8 （CCK-8） assay to observe fibroblast growth and determine the optimal intervention time. Alkaline phosphatase （ALP） activity was measured using the alkaline phosphatase assay. Protein expression of osteocalcin （OCN）， osteopontin （OPN）， and runt-related transcription factor 2 （Runx2） was detected by Western blot. The mRNA expression levels of Wnt5a， β-catenin， and Dickkopf-1 （DKK-1） were measured by real-time quantitative polymerase chain reaction （Real-time PCR）. ResultsCompared with the blank group， each drug-containing serum group of Wulao Qisun prescription and the positive drug group inhibited the proliferation of AS fibroblasts and reduced ALP expression （P<0.01）. Compared with the blank group， the low drug-containing serum group of Wulao Qisun prescription downregulated β-catenin mRNA expression （P<0.05）. The medium and high drug-containing serum groups and the positive drug group significantly downregulated Wnt5a and β-catenin mRNA expression （P<0.05， P<0.01）， with the positive drug group showing the most pronounced effect （P<0.01）. The high drug-containing serum group and the positive drug group significantly upregulated DKK-1 mRNA expression （P<0.01）. Compared with the blank group， the low drug-containing serum group of Wulao Qisun prescription inhibited the expression of OPN and Runx2 proteins （P<0.05， P<0.01）， while the medium and high drug-containing serum groups and the positive drug group inhibited the expression of OCN， OPN， and Runx2 proteins （P<0.05， P<0.01）. ConclusionThe Wulao Qisun prescription can inhibit the proliferation and osteogenic differentiation of AS fibroblasts， thereby delaying the formation of pathological new bone in AS. The possible mechanism involves the regulation of Wnt/β-catenin-related gene expression， further inhibiting the transcription of downstream target genes.

BACKGROUND:Rev-erbα is involved in the regulation of inflammation,but pharmacological activation of Rev-erbα increases the risk for cardiovascular diseases.To reduce the relevant risk,an exploration on SR9009,a Rev-erbα agonist,combined with other drugs to relieve inflammation in skeletal myoblasts was conducted,laying the theoretical foundation for the treatment of inflammation-associated skeletal muscle atrophy. OBJECTIVE:To investigate the relationship of SR9009,indolepropionic acid and nuclear factor-κB signaling pathways in lipopolysaccharide-induced C2C12 myoblasts. METHODS:(1)C2C12 myoblasts were induced to differentiate in the presence of lipopolysaccharide(1 μg/mL).RNA-seq and KEGG pathway analysis were used to study signaling pathways.(2)C2C12 myoblast viability was assessed using the cell counting kit-8 assay to determine optimal concentrations of indolepropionic acid.Subsequently,cells were categorized into control group,lipopolysaccharide(1 μg/mL)group,SR9009(10 μmol/L)+lipopolysaccharide group,indolepropionic acid(80μmol/L)+lipopolysaccharide group,and SR9009+indolepropionic acid+lipopolysaccharide group.ELISA was employed to measure protein expression levels of interleukin-6 in the cultured supernatant.Real-time quantitative PCR were employed to measure mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Western blot assay were employed to measure protein expression levels of NF-κB p65 and p-NF-κB p65.(3)After Rev-erbα was knocked down by siRNA,knockdown efficiency was assessed by RT-qPCR.And mRNA levels of interleukin-6 and tumor necrosis factor α were also measured. RESULTS AND CONCLUSION:Compared with the blank control group,lipopolysaccharide time-dependently inhibited myofibroblast fusion to form myotubes,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were elevated,and the level of interleukin-6 in the cell supernatant was significantly increased.The results of KEGG pathway showed that the nuclear factor-κB signaling pathway was activated by lipopolysaccharide.Indolepropionic acid exhibited significant suppression of C2C12 myoblasts viability when its concentration exceeded 80 μmol/L.Indolepropionic acid and SR9009 inhibited the activation of NF-κB signaling pathway,thereby played an anti-inflammatory role,and suppressed the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Compared with the lipopolysaccharide group,the ratio of p-NF-κB p65/NF-κB p65 protein expression were downregulated.SR9009 combined with indolepropionic acid notably reduced lipopolysaccharide-induced inflammation,further downregulated the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.The ratio of p-NF-κB p65/NF-κB p65 protein expression was significantly lower than that in the SR9009+lipopolysaccharide group or indolepropionic acid+lipopolysaccharide group.Rev-erbα increases time-dependently with lipopolysaccharide induction.The knockdown efficiency of Rev-erbα by siRNA reached over 58%,and lipopolysaccharide was added after Rev-erbα was successfully knocked down.Compared with the lipopolysaccharide group,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were significantly up-regulated.These results conclude that Rev-erbα may act as a promising pharmacological target to reduce inflammation.SR9009 targeted activation of Rev-erbα combined with indolepropionic acid significantly inhibits the nuclear factor-κB signaling pathway and attenuates the inflammatory response of C2C12 myofibroblasts.Moreover,the combined anti-inflammatory effect is superior to that of the intervention alone.

Early correction of childhood malocclusion is timely managing morphological, structural, and functional abnormalities at different dentomaxillofacial developmental stages. The selection of appropriate imaging examination and comprehensive radiological diagnosis and analysis play an important role in early correction of childhood malocclusion. This expert consensus is a collaborative effort by multidisciplinary experts in dentistry across the nation based on the current clinical evidence, aiming to provide general guidance on appropriate imaging examination selection, comprehensive and accurate imaging assessment for early orthodontic treatment patients.
Humans ; Malocclusion/diagnostic imaging* ; Child ; Consensus

During the Jin dynasty,the two major academic schools of Hejian and Yishui emerged in northern China.At this time,the Song dynasty migrated southward,accompanied by the southward movement of the Han ethnic culture and economic center.The dissemination of medicine also showed a trend of spreading from the north to the south.The late Yuan dynasty was an important period for the academic dissemination of the Yishui school to the south.On the one hand,ZHU Danxi,GE Yinglei,HUA Shou,and other clinicians studied the academic works of LI Dongyuan,comprehended his academic ideas,and further disseminated them through their disciples and Confucian scholars.On the other hand,academic works such as Jisheng Bacui and Weisheng Baojian carrying the study of Yishui were published in the south,playing an essential role in disseminating Yishui's study in the south.The dissemination of Yishui's learning to the south adopted a combination of book learning and mentorship,effectively breaking down the academic barriers between the Hejian and Yishui schools.The network formed by the interaction between medical scholars and Confucian scholars was an essential medium for academic dissemination.The study of Yishui was transmitted to the south and integrated with the original spleen and stomach theory in the south,promoting the further development of traditional Chinese medicine spleen and stomach theory.

During the Jin dynasty,the two major academic schools of Hejian and Yishui emerged in northern China.At this time,the Song dynasty migrated southward,accompanied by the southward movement of the Han ethnic culture and economic center.The dissemination of medicine also showed a trend of spreading from the north to the south.The late Yuan dynasty was an important period for the academic dissemination of the Yishui school to the south.On the one hand,ZHU Danxi,GE Yinglei,HUA Shou,and other clinicians studied the academic works of LI Dongyuan,comprehended his academic ideas,and further disseminated them through their disciples and Confucian scholars.On the other hand,academic works such as Jisheng Bacui and Weisheng Baojian carrying the study of Yishui were published in the south,playing an essential role in disseminating Yishui's study in the south.The dissemination of Yishui's learning to the south adopted a combination of book learning and mentorship,effectively breaking down the academic barriers between the Hejian and Yishui schools.The network formed by the interaction between medical scholars and Confucian scholars was an essential medium for academic dissemination.The study of Yishui was transmitted to the south and integrated with the original spleen and stomach theory in the south,promoting the further development of traditional Chinese medicine spleen and stomach theory.

During the Jin dynasty,the two major academic schools of Hejian and Yishui emerged in northern China.At this time,the Song dynasty migrated southward,accompanied by the southward movement of the Han ethnic culture and economic center.The dissemination of medicine also showed a trend of spreading from the north to the south.The late Yuan dynasty was an important period for the academic dissemination of the Yishui school to the south.On the one hand,ZHU Danxi,GE Yinglei,HUA Shou,and other clinicians studied the academic works of LI Dongyuan,comprehended his academic ideas,and further disseminated them through their disciples and Confucian scholars.On the other hand,academic works such as Jisheng Bacui and Weisheng Baojian carrying the study of Yishui were published in the south,playing an essential role in disseminating Yishui's study in the south.The dissemination of Yishui's learning to the south adopted a combination of book learning and mentorship,effectively breaking down the academic barriers between the Hejian and Yishui schools.The network formed by the interaction between medical scholars and Confucian scholars was an essential medium for academic dissemination.The study of Yishui was transmitted to the south and integrated with the original spleen and stomach theory in the south,promoting the further development of traditional Chinese medicine spleen and stomach theory.

During the Jin dynasty,the two major academic schools of Hejian and Yishui emerged in northern China.At this time,the Song dynasty migrated southward,accompanied by the southward movement of the Han ethnic culture and economic center.The dissemination of medicine also showed a trend of spreading from the north to the south.The late Yuan dynasty was an important period for the academic dissemination of the Yishui school to the south.On the one hand,ZHU Danxi,GE Yinglei,HUA Shou,and other clinicians studied the academic works of LI Dongyuan,comprehended his academic ideas,and further disseminated them through their disciples and Confucian scholars.On the other hand,academic works such as Jisheng Bacui and Weisheng Baojian carrying the study of Yishui were published in the south,playing an essential role in disseminating Yishui's study in the south.The dissemination of Yishui's learning to the south adopted a combination of book learning and mentorship,effectively breaking down the academic barriers between the Hejian and Yishui schools.The network formed by the interaction between medical scholars and Confucian scholars was an essential medium for academic dissemination.The study of Yishui was transmitted to the south and integrated with the original spleen and stomach theory in the south,promoting the further development of traditional Chinese medicine spleen and stomach theory.

During the Jin dynasty,the two major academic schools of Hejian and Yishui emerged in northern China.At this time,the Song dynasty migrated southward,accompanied by the southward movement of the Han ethnic culture and economic center.The dissemination of medicine also showed a trend of spreading from the north to the south.The late Yuan dynasty was an important period for the academic dissemination of the Yishui school to the south.On the one hand,ZHU Danxi,GE Yinglei,HUA Shou,and other clinicians studied the academic works of LI Dongyuan,comprehended his academic ideas,and further disseminated them through their disciples and Confucian scholars.On the other hand,academic works such as Jisheng Bacui and Weisheng Baojian carrying the study of Yishui were published in the south,playing an essential role in disseminating Yishui's study in the south.The dissemination of Yishui's learning to the south adopted a combination of book learning and mentorship,effectively breaking down the academic barriers between the Hejian and Yishui schools.The network formed by the interaction between medical scholars and Confucian scholars was an essential medium for academic dissemination.The study of Yishui was transmitted to the south and integrated with the original spleen and stomach theory in the south,promoting the further development of traditional Chinese medicine spleen and stomach theory.

During the Jin dynasty,the two major academic schools of Hejian and Yishui emerged in northern China.At this time,the Song dynasty migrated southward,accompanied by the southward movement of the Han ethnic culture and economic center.The dissemination of medicine also showed a trend of spreading from the north to the south.The late Yuan dynasty was an important period for the academic dissemination of the Yishui school to the south.On the one hand,ZHU Danxi,GE Yinglei,HUA Shou,and other clinicians studied the academic works of LI Dongyuan,comprehended his academic ideas,and further disseminated them through their disciples and Confucian scholars.On the other hand,academic works such as Jisheng Bacui and Weisheng Baojian carrying the study of Yishui were published in the south,playing an essential role in disseminating Yishui's study in the south.The dissemination of Yishui's learning to the south adopted a combination of book learning and mentorship,effectively breaking down the academic barriers between the Hejian and Yishui schools.The network formed by the interaction between medical scholars and Confucian scholars was an essential medium for academic dissemination.The study of Yishui was transmitted to the south and integrated with the original spleen and stomach theory in the south,promoting the further development of traditional Chinese medicine spleen and stomach theory.

During the Jin dynasty,the two major academic schools of Hejian and Yishui emerged in northern China.At this time,the Song dynasty migrated southward,accompanied by the southward movement of the Han ethnic culture and economic center.The dissemination of medicine also showed a trend of spreading from the north to the south.The late Yuan dynasty was an important period for the academic dissemination of the Yishui school to the south.On the one hand,ZHU Danxi,GE Yinglei,HUA Shou,and other clinicians studied the academic works of LI Dongyuan,comprehended his academic ideas,and further disseminated them through their disciples and Confucian scholars.On the other hand,academic works such as Jisheng Bacui and Weisheng Baojian carrying the study of Yishui were published in the south,playing an essential role in disseminating Yishui's study in the south.The dissemination of Yishui's learning to the south adopted a combination of book learning and mentorship,effectively breaking down the academic barriers between the Hejian and Yishui schools.The network formed by the interaction between medical scholars and Confucian scholars was an essential medium for academic dissemination.The study of Yishui was transmitted to the south and integrated with the original spleen and stomach theory in the south,promoting the further development of traditional Chinese medicine spleen and stomach theory.