Objective:To explore the therapeutic mechanism of moxibustion in Crohn disease(CD)-associated intestinal fibrosis by observing its effects on the angiotensin-converting enzyme(ACE)/angiotensin Ⅱ(Ang Ⅱ)/angiotensin Ⅱ type 1 receptor(AT1R)axis in CD mouse models.Methods:Six randomly selected male C57BL/6 mice were assigned to a normal group,while the remaining mice were administered 0.1 mL of 2,4,6-trinitrobenzene sulfonic acid via enema to establish a CD intestinal fibrosis model.After successful modeling,the mice were randomly divided into a model group,a moxibustion group,and a Western medication group,with 6 rats in each group.The normal group and the model group only received grabbing without intervention.In the moxibustion group,mild moxibustion was applied to Qihai(CV6)and bilateral Tianshu(ST25)once a day for 10 min each time over 7 consecutive days.The Western medication group was administered mesalazine suspension via oral gavage once a day for 7 consecutive days.At the end of the intervention,the general condition,disease activity index(DAI)score,and gross colon score of mice in each group were evaluated.Hematoxylin-eosin staining was used to observe and score the histological changes in the colon tissue in each group.Masson staining was used to observe colonic fibrosis and the ratio of collagen-positive areas was analyzed;the expression of Ang Ⅱ in the colon tissue was detected by the enzyme-linked immunosorbent assay;immunohistochemistry and real-time quantitative reverse transcription polymerase chain reaction were used to detect the protein and mRNA expression of ACE and AT1R in the colon tissue,respectively;Western blotting was used to detect the expression of transforming growth factor(TGF)-β1 and connective tissue growth factor(CTGF)in the colon tissue.Results:Compared to the normal group,the DAI score,gross colon score,colonic histological score,collagen-positive area ratio,ACE protein and mRNA,Ang Ⅱ protein,AT1R protein and mRNA,TGF-β1 protein,and CTGF protein in the colon tissue in the model group increased significantly(P<0.01).In contrast,the above indicators in both the moxibustion group and the Western medication group reduced significantly compared to the model group(P<0.01 or P<0.05).There was no statistical difference in these indicators between the moxibustion group and the Western medication group(P>0.05).Conclusion:Moxibustion can alleviate intestinal fibrosis in CD mice,and its therapeutic mechanism may be associated with the regulation of colonic ACE/AngⅡ/AT1R axis.

Objective:To explore the therapeutic mechanism of moxibustion in Crohn disease(CD)-associated intestinal fibrosis by observing its effects on the angiotensin-converting enzyme(ACE)/angiotensin Ⅱ(Ang Ⅱ)/angiotensin Ⅱ type 1 receptor(AT1R)axis in CD mouse models.Methods:Six randomly selected male C57BL/6 mice were assigned to a normal group,while the remaining mice were administered 0.1 mL of 2,4,6-trinitrobenzene sulfonic acid via enema to establish a CD intestinal fibrosis model.After successful modeling,the mice were randomly divided into a model group,a moxibustion group,and a Western medication group,with 6 rats in each group.The normal group and the model group only received grabbing without intervention.In the moxibustion group,mild moxibustion was applied to Qihai(CV6)and bilateral Tianshu(ST25)once a day for 10 min each time over 7 consecutive days.The Western medication group was administered mesalazine suspension via oral gavage once a day for 7 consecutive days.At the end of the intervention,the general condition,disease activity index(DAI)score,and gross colon score of mice in each group were evaluated.Hematoxylin-eosin staining was used to observe and score the histological changes in the colon tissue in each group.Masson staining was used to observe colonic fibrosis and the ratio of collagen-positive areas was analyzed;the expression of Ang Ⅱ in the colon tissue was detected by the enzyme-linked immunosorbent assay;immunohistochemistry and real-time quantitative reverse transcription polymerase chain reaction were used to detect the protein and mRNA expression of ACE and AT1R in the colon tissue,respectively;Western blotting was used to detect the expression of transforming growth factor(TGF)-β1 and connective tissue growth factor(CTGF)in the colon tissue.Results:Compared to the normal group,the DAI score,gross colon score,colonic histological score,collagen-positive area ratio,ACE protein and mRNA,Ang Ⅱ protein,AT1R protein and mRNA,TGF-β1 protein,and CTGF protein in the colon tissue in the model group increased significantly(P<0.01).In contrast,the above indicators in both the moxibustion group and the Western medication group reduced significantly compared to the model group(P<0.01 or P<0.05).There was no statistical difference in these indicators between the moxibustion group and the Western medication group(P>0.05).Conclusion:Moxibustion can alleviate intestinal fibrosis in CD mice,and its therapeutic mechanism may be associated with the regulation of colonic ACE/AngⅡ/AT1R axis.

Bones possess metabolic activity, with their homeostasis maintained by bone resorption and bone formation mediated by osteoclasts and osteoblasts. By measuring bone metabolism markers, the overall state of bone metabolism and dynamic changes in systemic bone tissue can be reflected. Traditional bone turnover markers, including alkaline phosphatase, bonespecific alkaline phosphatase, procollagen type 1 N-terminal propeptide, procollagen type 1 C-terminal propeptide, osteocalcin, c-terminal telopeptides of type 1 collagen(CTX) and its subtype β-CTX, n-terminal telopeptides of type 1 collagen, have been widely used in clinical practice but still have limitations in terms of stability, diagnostic reliability, and specific reflection of bone sites. Recently, novel bone turnover markers like microRNA, C-X-C chemokine ligand 12, Gelsolin, Annexin A2, sclerostin, Dickkopf-related protein 1, and citrate have garnered significant attention. This article endeavors to conduct a review of the production, mechanism of action, detection methods, diagnostic value, and application prospects of new bone turnover markers in osteoporosis, thereby offering novel ideas for the prevention, diagnosis, and treatment of osteoporosis.

Bones possess metabolic activity, with their homeostasis maintained by bone resorption and bone formation mediated by osteoclasts and osteoblasts. By measuring bone metabolism markers, the overall state of bone metabolism and dynamic changes in systemic bone tissue can be reflected. Traditional bone turnover markers, including alkaline phosphatase, bonespecific alkaline phosphatase, procollagen type 1 N-terminal propeptide, procollagen type 1 C-terminal propeptide, osteocalcin, c-terminal telopeptides of type 1 collagen(CTX) and its subtype β-CTX, n-terminal telopeptides of type 1 collagen, have been widely used in clinical practice but still have limitations in terms of stability, diagnostic reliability, and specific reflection of bone sites. Recently, novel bone turnover markers like microRNA, C-X-C chemokine ligand 12, Gelsolin, Annexin A2, sclerostin, Dickkopf-related protein 1, and citrate have garnered significant attention. This article endeavors to conduct a review of the production, mechanism of action, detection methods, diagnostic value, and application prospects of new bone turnover markers in osteoporosis, thereby offering novel ideas for the prevention, diagnosis, and treatment of osteoporosis.

Objective: To explore the effects of lncRNA HULC on hepatocellular carcinoma (HCC) growth by down-regulating miR-29. Methods: The expression levels of HULC and miR-29 in HCC tissues and cells were detected by real-time quantitative PCR (RT-qPCR), and the correlation analysis was performed. After HCC cells were transfected with HULC overexpressed plasmid or siRNA, the expressions of miR-29 and its target gene SETDB1 were determinate by RT-qPCR. According to the bioinformatic prediction of the miR-29 binding site in the HULC sequence, the report gene plasmids were constructed. The HCC cells were co-transfected with miR-29 mimics or miR-29 inhibitor, and the HULC targeted regulation of miR-29 was verified by dual luciferase reporter assay. The effect of miR-29 on the HULC-mediated proliferation in HCC cells was detected by cell count kit 8 (CCK-8) experiment. Expression of tumor proliferation antigen Ki-67 was detected by RT-qPCR.The Hep3B cells were inoculated in mice and miR-29 mimics and miR-29 negative control (NC) further injected into the lesions. The tumor volume was observed, and the expressions of tumor proliferation antigen ki-67 in tumor tissues were detected by immunohistochemical staining. Results: The expression of HULC was significantly up-regulated while the expression of miR-29 was significantly down-regulated in HCC tissues and cells (P<0.01). The level of HULC was negatively correlated with miR-29 in tumor tissues (r=-0.754, P<0.01) and HCC cells (r=-0.865, P<0.05). The in vitro experiments showed that, compared with the blank control group, the expression of miR-29 in HULC overexpressed Huh7 cells was significantly reduced, while the mRNA level of miR-29 target gene SETDB1 was increased (P<0.01). The expression of miR-29 was significantly increased in HULC deleted Hep3B cells, while the mRNA expression of SETDB1 was decreased (P<0.01). Double luciferase reporter gene assay showed that miR-29 mimics significantly inhibited the luciferase activity of Hep3B cells transfected with HULC wide type (psi-HULC-WT) plasmid but had no effect on Hep3B cells transfected with mutant plasmid (psi-HULC-Mut). However, the miR-29 inhibitor antagonized the inhibitory effect of miR-29 mimics on luciferase activity of psi-HULC-WT (P<0.01). Cell proliferation experiments showed that, compared with the control group, the proliferation ability of miR-29 mimics overexpressed Huh7 cells was significantly reduced.After 24, 48 and 72 hours of treatment, the proliferation rates of Huh7 cells in the HULC overexpressed group were (43.87±3.82)%, (83.45±7.46)% and (123.34±8.67)%, respectively, significantly higher than (13.45±1.77)%, (23.54±1.37)% and (38.21±2.09)% of control group (P<0.05). After treatment for 48 and 72 hours, the proliferation rates of miR-29 mimics transfected Huh7 cells were (57.10±1.94)% and (73.76±3.46)%, respectively, significantly lower than (83.45±7.46)% and (123.34±8.67)% of control group (P<0.05). After treatment for 48 and 72 hours, the proliferation rates of Huh7 cells transfected with miR-29 mimics and miR-29 inhibitor group were (76.45±3.24)% and (89.37±4.37)%, respectively, significant higher than (57.10%±1.94)% and (73.76±3.46)% of the control group (P<0.05). After 48 h transfection, the expression of Ki-67 in Huh7 transfected with miR-29 mimics was significantly inhibited compared with the control group (P<0.01). However, the expression of Ki-67 mRNA was increased in Huh7 cells transfected with miR-29 inhibitor (P<0.01). The results of in vivo experiments showed that the tumor volumes of the control group, miR-29 mimics group and miR-29 mimics + miR-29 inhibitors group were (504.0±19.6) mm³, (310.0±24.3) mm³ and (483.7±21.2) mm³, respectively. Injection of miR-29 mimics reduced while miR-29 inhibitor promoted tumorigenesis ability of Huh7 in nude mice (P<0.01). The immunohistochemical staining showed that the average optical density values of Ki-67 protein in tumor tissues of the control group, miR-29 mimics group and miR-29 analogue+ miR-29 inhibitor group were 0.65±0.08, 0.36±0.07 and 0.56±0.06, respectively. The expression level of Ki-67 protein in miR-29 mimics group was significantly reduced (P<0.01) while increased in the miR-29 mimics+ miR-29 inhibitor group (P<0.01). Conclusion LncRNA HULC promotes HCC growth by down-regulating miR-29.

Objective To explore the effects of lncRNA HULC on hepatocellular carcinoma (HCC) growth by down?regulating miR?29. Methods The expression levels of HULC and miR?29 in HCC tissues and cells were detected by real?time quantitative PCR ( RT?qPCR ), and the correlation analysis was performed. After HCC cells were transfected with HULC overexpressed plasmid or siRNA, the expressions of miR?29 and its target gene SETDB1 were determinate by RT?qPCR. According to the bioinformatic prediction of the miR?29 binding site in the HULC sequence, the report gene plasmids were constructed.The HCC cells were co?transfected with miR?29 mimics or miR?29 inhibitor, and the HULC targeted regulation of miR?29 was verified by dual luciferase reporter assay. The effect of miR?29 on the HULC?mediated proliferation in HCC cells was detected by cell count kit 8 ( CCK?8) experiment. Expression of tumor proliferation antigen Ki?67 was detected by RT?qPCR.The Hep3B cells were inoculated in mice and miR?29 mimics and miR?29 negative control (NC) further injected into the lesions. The tumor volume was observed, and the expressions of tumor proliferation antigen ki?67 in tumor tissues were detected by immunohistochemical staining. Results The expression of HULC was significantly up?regulated while the expression of miR?29 was significantly down?regulated in HCC tissues and cells (P＜0.01). The level of HULC was negatively correlated with miR?29 in tumor tissues (r＝－0.754, P＜0.01) and HCC cells ( r＝－0.865, P＜0.05).The in vitro experiments showed that, compared with the blank control group, the expression of miR?29 in HULC overexpressed Huh7 cells was significantly reduced, while the mRNA level of miR?29 target gene SETDB1 was increased ( P＜0.01). The expression of miR?29 was significantly increased in HULC deleted Hep3B cells, while the mRNA expression of SETDB1 was decreased (P＜0.01). Double luciferase reporter gene assay showed that miR?29 mimics significantly inhibited the luciferase activity of Hep3B cells transfected with HULC wide type ( psi?HULC?WT) plasmid but had no effect on Hep3B cells transfected with mutant plasmid ( psi?HULC?Mut). However, the miR?29 inhibitor antagonized the inhibitory effect of miR?29 mimics on luciferase activity of psi?HULC?WT (P＜0.01).Cell proliferation experiments showed that, compared with the control group, the proliferation ability of miR?29 mimics overexpressed Huh7 cells was significantly reduced.After 24, 48 and 72 hours of treatment, the proliferation rates of Huh7 cells in the HULC overexpressed group were (43.87± 3.82)%, (83.45±7.46)% and (123.34±8.67)%, respectively, significantly higher than (13.45±1.77)%, (23.54±1.37)% and (38.21±2.09)% of control group (P＜0.05).After treatment for 48 and 72 hours, the proliferation rates of miR?29 mimics transfected Huh7 cells were ( 57.10 ± 1.94)% and ( 73.76± 3.46)%, respectively, significantly lower than (83.45±7.46)% and ( 123.34±8.67)% of control group ( P＜0.05). After treatment for 48 and 72 hours, the proliferation rates of Huh7 cells transfected with miR?29 mimics and miR?29 inhibitor group were (76.45± 3.24)% and ( 89.37± 4.37)%, respectively, significant higher than (57.10%±1.94)% and ( 73.76 ± 3.46)% of the control group ( P＜0.05). After 48 h transfection, the expression of Ki?67 in Huh7 transfected with miR?29 mimics was significantly inhibited compared with the control group (P＜0.01). However, the expression of Ki?67 mRNA was increased in Huh7 cells transfected with miR?29 inhibitor (P＜0.01). The results of in vivo experiments showed that the tumor volumes of the control group, miR?29 mimics group and miR?29 mimics + miR?29 inhibitors group were ( 504.0± 19.6) mm3, (310.0±24.3) mm3 and (483.7±21.2) mm3, respectively. Injection of miR?29 mimics reduced while miR?29 inhibitor promoted tumorigenesis ability of Huh7 in nude mice (P＜0.01). The immunohistochemical staining showed that the average optical density values of Ki?67 protein in tumor tissues of the control group, miR?29 mimics group and miR?29 analogue+miR?29 inhibitor group were 0.65±0.08, 0.36±0.07 and 0.56±0.06, respectively. The expression level of Ki?67 protein in miR?29 mimics group was significantly reduced ( P＜0.01) while increased in the miR?29 mimics+miR?29 inhibitor group ( P＜0.01). Conclusion LncRNA HULC promotes HCC growth by down?regulating miR?29.

Objective To explore the effects of lncRNA HULC on hepatocellular carcinoma (HCC) growth by down?regulating miR?29. Methods The expression levels of HULC and miR?29 in HCC tissues and cells were detected by real?time quantitative PCR ( RT?qPCR ), and the correlation analysis was performed. After HCC cells were transfected with HULC overexpressed plasmid or siRNA, the expressions of miR?29 and its target gene SETDB1 were determinate by RT?qPCR. According to the bioinformatic prediction of the miR?29 binding site in the HULC sequence, the report gene plasmids were constructed.The HCC cells were co?transfected with miR?29 mimics or miR?29 inhibitor, and the HULC targeted regulation of miR?29 was verified by dual luciferase reporter assay. The effect of miR?29 on the HULC?mediated proliferation in HCC cells was detected by cell count kit 8 ( CCK?8) experiment. Expression of tumor proliferation antigen Ki?67 was detected by RT?qPCR.The Hep3B cells were inoculated in mice and miR?29 mimics and miR?29 negative control (NC) further injected into the lesions. The tumor volume was observed, and the expressions of tumor proliferation antigen ki?67 in tumor tissues were detected by immunohistochemical staining. Results The expression of HULC was significantly up?regulated while the expression of miR?29 was significantly down?regulated in HCC tissues and cells (P＜0.01). The level of HULC was negatively correlated with miR?29 in tumor tissues (r＝－0.754, P＜0.01) and HCC cells ( r＝－0.865, P＜0.05).The in vitro experiments showed that, compared with the blank control group, the expression of miR?29 in HULC overexpressed Huh7 cells was significantly reduced, while the mRNA level of miR?29 target gene SETDB1 was increased ( P＜0.01). The expression of miR?29 was significantly increased in HULC deleted Hep3B cells, while the mRNA expression of SETDB1 was decreased (P＜0.01). Double luciferase reporter gene assay showed that miR?29 mimics significantly inhibited the luciferase activity of Hep3B cells transfected with HULC wide type ( psi?HULC?WT) plasmid but had no effect on Hep3B cells transfected with mutant plasmid ( psi?HULC?Mut). However, the miR?29 inhibitor antagonized the inhibitory effect of miR?29 mimics on luciferase activity of psi?HULC?WT (P＜0.01).Cell proliferation experiments showed that, compared with the control group, the proliferation ability of miR?29 mimics overexpressed Huh7 cells was significantly reduced.After 24, 48 and 72 hours of treatment, the proliferation rates of Huh7 cells in the HULC overexpressed group were (43.87± 3.82)%, (83.45±7.46)% and (123.34±8.67)%, respectively, significantly higher than (13.45±1.77)%, (23.54±1.37)% and (38.21±2.09)% of control group (P＜0.05).After treatment for 48 and 72 hours, the proliferation rates of miR?29 mimics transfected Huh7 cells were ( 57.10 ± 1.94)% and ( 73.76± 3.46)%, respectively, significantly lower than (83.45±7.46)% and ( 123.34±8.67)% of control group ( P＜0.05). After treatment for 48 and 72 hours, the proliferation rates of Huh7 cells transfected with miR?29 mimics and miR?29 inhibitor group were (76.45± 3.24)% and ( 89.37± 4.37)%, respectively, significant higher than (57.10%±1.94)% and ( 73.76 ± 3.46)% of the control group ( P＜0.05). After 48 h transfection, the expression of Ki?67 in Huh7 transfected with miR?29 mimics was significantly inhibited compared with the control group (P＜0.01). However, the expression of Ki?67 mRNA was increased in Huh7 cells transfected with miR?29 inhibitor (P＜0.01). The results of in vivo experiments showed that the tumor volumes of the control group, miR?29 mimics group and miR?29 mimics + miR?29 inhibitors group were ( 504.0± 19.6) mm3, (310.0±24.3) mm3 and (483.7±21.2) mm3, respectively. Injection of miR?29 mimics reduced while miR?29 inhibitor promoted tumorigenesis ability of Huh7 in nude mice (P＜0.01). The immunohistochemical staining showed that the average optical density values of Ki?67 protein in tumor tissues of the control group, miR?29 mimics group and miR?29 analogue+miR?29 inhibitor group were 0.65±0.08, 0.36±0.07 and 0.56±0.06, respectively. The expression level of Ki?67 protein in miR?29 mimics group was significantly reduced ( P＜0.01) while increased in the miR?29 mimics+miR?29 inhibitor group ( P＜0.01). Conclusion LncRNA HULC promotes HCC growth by down?regulating miR?29.