Objective To explore the effects of eugenol combined with traditional Chinese medicine Wuxing music(EUG+MUSIC)on skin inflammation,behavioral characteristics,hormone and neurotransmitter levels,and the intestinal microbiota in a mouse model of 2,4-dinitrofluorobenzene(DNFB)-induced atopic dermatitis(AD).Methods A mouse model of AD was established by induction with DNFB for 21 consecutive days;throughout the study,changes in dorsal skin lesions and scratching behavior were recorded in mice and changes in dorsal skin lesions and pruritus were recorded.Mice underwent open-field,elevated 0 maze,tail-suspension,and forced-swimming tests on days 19 and 20.The mice were then sacrificed on day 21 and the organ index was calculated.Serum levels of immunoglobulin E,interleukin(IL)-4,and IL-10 were determined by enzyme-linked immunosorbent assay.Pathological changes in back-skin sections were detected by hematoxylin-eosin(HE)and toluidine blue staining,and brain levels of corticosterone,dopamine,and serotonin were measured.The composition of the intestinal microbiota was analyzed by 16S rRNA gene sequencing.Results The skin lesion score and skin inflammatory cell infiltration were significantly reduced in the EUG+MUSIC group.Regarding the behavioral tests,mice in the EUG+MUSIC group showed more active locomotion,an increased desire to explore unfamiliar environments,and increased struggling time in stressful situations.Brain levels of serotonin,dopamine,and corticosterone were also increased in the EUG+MUSIC group.16S rRNA gene sequencing showed that DNFB affected the diversity of the intestinal microbiota in AD mice,while EUG+MUSIC treatment remodeled the gut microbial composition,including increasing the relative abundance of gut microbes such as Ligilactobacillus and Lachnospiraceae and decreasing Parabacteroides and Bacteroides spp.Spearman's analysis showed significant correlations among altered gut microbiota,AD-associated skin manifestations,and anxiety-and depressive-like behaviors arising from comorbidities.Conclusions Eugenol combined with traditional Chinese medicine five-elements music therapy may have a positive effect on comorbid anxiety-and depression-like behaviors by regulating the intestinal microbiota and improving the behavioral performance in mice with AD,thus providing new ideas and method for improving AD and its associated anxiety-and depressive behaviors.

Objective:To study the methylation level of miR-5194 promoter in bladder cancer tissues, and explore the effects of miR-5194 on the proliferation and migration of bladder cancer cells by targeting p21-activated protein kinase 2 (PAK2).Methods:The methylation level of miR-5194 promoter in bladder cancer tissues was analyzed using MethHC database. Real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of miR-5194 in bladder cancer MGH-U3, EJ, J82, and UMUC3 cells. 5-aza-2′-deoxycytidine (5-Aza-CdR) was used to treat bladder cancer cell lines, and RT-qPCR was used to detect the changes in the expression of miR-5194 in bladder cancer cell lines after 5-Aza-CdR treatment. UMUC3 cells were divided into miR-5194 group and NC group, and miR-5194 or miR-NC were transfected into UMUC3 cells, respectively. Colony formation assay and scratch assay were used to detect the effect of overexpression of miR-5194 on the proliferation and migration of UMUC3 cells. The bioinformatics tool miRGator and dual-luciferase reporter gene experiments verified the targeting relationship between miR-5194 and PAK2. The effect of overexpression of miR-5194 on the expression of PAK2 mRNA in UMUC3 cells was detected by RT-qPCR. The effect of overexpression of miR-5194 on the expression of PAK2 protein, proliferation-related proteins (CDK1, Cyclin B) and migration-related proteins (FOXC2, E47) in UMUC3 cells was detected by Western blotting. The measurement data were expressed as mean ± standard deviation ( ± s), the independent sample t-test was used for comparison between two groups, and one-way analysis of variance was used for comparison among multiple groups. Results:The methylation level of miR-5194 promoter in bladder cancer tissues was significantly higher than that in adjacent tissues ( P<0.01). Compared with the immortalized bladder epithelial cells SV-HUC-1, the expression of miR-5194 in bladder cancer cells was significantly down-regulated ( P<0.01). After 5-Aza-CdR treatment, the expression of miR-5194 in bladder cancer cells was significantly increased ( P<0.01). The number of colonies in miR-5194 group and NC group were 31.30 ± 8.09 and 99.98 ± 10.53, respectively, and the proliferation ability of UMUC3 cells in miR-5194 group was weakened ( P<0.01). The migration rates of UMUC3 cells in miR-5194 group and NC group were (31.50 ± 7.17)% and (76.06 ± 4.86)%, respectively, and the migration ability of UMUC3 cells in miR-5194 group was weakened ( P<0.01). miR-5194 can target bind PAK2 gene ( P<0.01). The relative expression of PAK2 mRNA in UMUC3 cells of miR-5194 group and NC group were 1.02 ± 0.34 and 5.43 ± 0.76, respectively, and miR-5194 could negatively regulate the expression of PAK2 mRNA ( P<0.01). Compared with the NC group, the expression of PAK2 protein, the expression of proliferation-related proteins CDK1 and Cyclin B, and the expression of migration-related proteins FOXC2 and E47 were down-regulated in UMUC3 cells with miR-5194 overexpression. Conclusion:The methylation level of miR-5194 promoter in bladder cancer tissue was significantly increased, and miR-5194 inhibited the proliferation and migration of bladder cancer cells by targeting down-regulation of PAK2 expression in bladder cancer UMUC3 cells.

Objective:To compare the burden of benign prostatic hyperplasia（BPH）between China and the United States from 1990 to 2021.Methods:The prevalence，incidence，years lived with disability（YLD），and their age-standardized rates for BPH in China and the United States from 1990 to 2021 were obtained from the Global Burden of Disease Study 2021（GBD 2021）. The average annual percentage change（AAPC）of the age-standardized incidence rate（ASIR）and the age-standardized YLD rate（ASYR）was calculated using Joinpoint regression analysis. In addition，the YLD burden of BPH，prostate cancer，kidney cancer，bladder cancer，and three other urological diseases were compared between the two countries.Results:From 1990 to 2021，the number of BPH cases in China increased from 1.460 4 million to 3.244 5 million，the number of prevalent cases rose from 9.940 5 million to 23.111 2 million，and YLDs grew from 0.2 million person-years to 0.460 2 million person-years，with AAPCs of 2.63%，2.78%，and 2.75%，respectively. In 2021，the numbers of incident cases，prevalent cases，and YLDs were 0.577 9 million，4.930 3 million，and 0.095 9 million person-years in the United States，and 13.787 6 million，112.502 million，and 2.235 7 million person-years globally. China’s ASIR decreased from 363.07/100 000 to 299.14/100 000（AAPC -0.60%），and ASYR from 57.33/100 000 to 45.84/100 000（AAPC -0.70%），both of which were higher than those in the United States but lower than the global level. Age-specific analyses showed declining incidence and YLD rates across all age groups in China，while certain age groups in the United States demonstrated increasing trends. From 1990 to 2021，the proportion of YLDs attributable to BPH among seven urological diseases in China rose from 61.4% to 69.2%. In 2021，YLDs due to prostate cancer accounted for the highest proportion among seven urinary system diseases in the United States，reaching 54.5%. Projections indicate that although ASIR and ASYR in China will decline from 2022 to 2040，the absolute numbers of incident cases and YLDs are projected to continue to rise，reaching 4.97 million and 0.78 million，respectively，by 2040.Conclusions:Between 1990 and 2021，the number of incidence cases，prevalence cases，and YLDs of BPH in China increased markedly，while ASIR and ASYR declined. The disease burden of BPH remains substantial，with a higher proportion of YLDs among urological diseases compared with the United States. By 2040，the number of BPH cases and YLDs in China is projected to further increase，underscoring the need for greater public health attention.

Objective To explore the effects of eugenol combined with traditional Chinese medicine Wuxing music(EUG+MUSIC)on skin inflammation,behavioral characteristics,hormone and neurotransmitter levels,and the intestinal microbiota in a mouse model of 2,4-dinitrofluorobenzene(DNFB)-induced atopic dermatitis(AD).Methods A mouse model of AD was established by induction with DNFB for 21 consecutive days;throughout the study,changes in dorsal skin lesions and scratching behavior were recorded in mice and changes in dorsal skin lesions and pruritus were recorded.Mice underwent open-field,elevated 0 maze,tail-suspension,and forced-swimming tests on days 19 and 20.The mice were then sacrificed on day 21 and the organ index was calculated.Serum levels of immunoglobulin E,interleukin(IL)-4,and IL-10 were determined by enzyme-linked immunosorbent assay.Pathological changes in back-skin sections were detected by hematoxylin-eosin(HE)and toluidine blue staining,and brain levels of corticosterone,dopamine,and serotonin were measured.The composition of the intestinal microbiota was analyzed by 16S rRNA gene sequencing.Results The skin lesion score and skin inflammatory cell infiltration were significantly reduced in the EUG+MUSIC group.Regarding the behavioral tests,mice in the EUG+MUSIC group showed more active locomotion,an increased desire to explore unfamiliar environments,and increased struggling time in stressful situations.Brain levels of serotonin,dopamine,and corticosterone were also increased in the EUG+MUSIC group.16S rRNA gene sequencing showed that DNFB affected the diversity of the intestinal microbiota in AD mice,while EUG+MUSIC treatment remodeled the gut microbial composition,including increasing the relative abundance of gut microbes such as Ligilactobacillus and Lachnospiraceae and decreasing Parabacteroides and Bacteroides spp.Spearman's analysis showed significant correlations among altered gut microbiota,AD-associated skin manifestations,and anxiety-and depressive-like behaviors arising from comorbidities.Conclusions Eugenol combined with traditional Chinese medicine five-elements music therapy may have a positive effect on comorbid anxiety-and depression-like behaviors by regulating the intestinal microbiota and improving the behavioral performance in mice with AD,thus providing new ideas and method for improving AD and its associated anxiety-and depressive behaviors.

Objective:To compare the burden of benign prostatic hyperplasia（BPH）between China and the United States from 1990 to 2021.Methods:The prevalence，incidence，years lived with disability（YLD），and their age-standardized rates for BPH in China and the United States from 1990 to 2021 were obtained from the Global Burden of Disease Study 2021（GBD 2021）. The average annual percentage change（AAPC）of the age-standardized incidence rate（ASIR）and the age-standardized YLD rate（ASYR）was calculated using Joinpoint regression analysis. In addition，the YLD burden of BPH，prostate cancer，kidney cancer，bladder cancer，and three other urological diseases were compared between the two countries.Results:From 1990 to 2021，the number of BPH cases in China increased from 1.460 4 million to 3.244 5 million，the number of prevalent cases rose from 9.940 5 million to 23.111 2 million，and YLDs grew from 0.2 million person-years to 0.460 2 million person-years，with AAPCs of 2.63%，2.78%，and 2.75%，respectively. In 2021，the numbers of incident cases，prevalent cases，and YLDs were 0.577 9 million，4.930 3 million，and 0.095 9 million person-years in the United States，and 13.787 6 million，112.502 million，and 2.235 7 million person-years globally. China’s ASIR decreased from 363.07/100 000 to 299.14/100 000（AAPC -0.60%），and ASYR from 57.33/100 000 to 45.84/100 000（AAPC -0.70%），both of which were higher than those in the United States but lower than the global level. Age-specific analyses showed declining incidence and YLD rates across all age groups in China，while certain age groups in the United States demonstrated increasing trends. From 1990 to 2021，the proportion of YLDs attributable to BPH among seven urological diseases in China rose from 61.4% to 69.2%. In 2021，YLDs due to prostate cancer accounted for the highest proportion among seven urinary system diseases in the United States，reaching 54.5%. Projections indicate that although ASIR and ASYR in China will decline from 2022 to 2040，the absolute numbers of incident cases and YLDs are projected to continue to rise，reaching 4.97 million and 0.78 million，respectively，by 2040.Conclusions:Between 1990 and 2021，the number of incidence cases，prevalence cases，and YLDs of BPH in China increased markedly，while ASIR and ASYR declined. The disease burden of BPH remains substantial，with a higher proportion of YLDs among urological diseases compared with the United States. By 2040，the number of BPH cases and YLDs in China is projected to further increase，underscoring the need for greater public health attention.

Objective:By observing the effects of Calycosin on the proliferation and migration of human renal cancer 769-P cell, to explore the possible molecular mechanism of Calycosin against renal cancer.Methods:769-P cell were cultured with different concentrations of Calycosin [0, 12.5, 25, 50, 100, 200 μmol/L, dissolved in Dimethyl sulfoxide (DMSO)], and the effects of different concentrations of Calycosin on the viability of 769-P cell was detected by CCK8 method. The 769-P cell treated with 200 μmol/L Calycosin were used as the Calycosin group, and the 769-P cell treated with DMSO were used as the control group. The cell colony formation assay and cell scratch assay were used to detect the effects of Calycosin on the proliferation and migration of 769-P cell, respectively. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the effect of Calycosin on the expression of miRNA-1246 and chemokine receptor-4 (CXCR4) in 769-P cell. Western blotting method was used to detect the effects of Calycosin on the expression of CXCR4 and extracellular signal-regulated kinase (ERK) pathway proteins in 769-P cell. Measurement data were expressed as mean ± standard deviation ( ± s), and one-way ANOVA was used for comparison between multiple groups, while t-test was used for comparison between two groups. Results:After cultured with 0, 12.5, 25, 50, 100, and 200 μmol/L of Calycosin, the absorbance values of renal cancer 769-P cell were 0.99 ± 0.06, 0.74 ± 0.07, 0.60 ± 0.03, 0.55 ± 0.05, 0.40 ± 0.06, 0.21 ± 0.04, respectively; compared with 0 μmol/L, the Calycosin could reduce the survival rate of 769-P cell ( P<0.05). The number of clones of 769-P cell in the control group and the Calycosin group was 109.80 ± 13.19 and 60.66 ± 11.22, respectively, and the number of clones of the 769-P cell in the Calycosin group was decreased, the difference was statistically significant ( t=5.67, P<0.01). The relative migration rates of 769-P cell in the control group and the Calycosin group were (43.13 ± 3.82)% and (14.27 ± 3.25)%, respectively, after the 769-P cell were treated with Calycosin, the cell migration ability was weakened ( t=5.71, P<0.05). The relative expression levels of miRNA-1246 in 769-P cell of the control group and the Calycosin group was 1.03 ± 0.12 and 6.99 ± 1.84, respectively, and the relative expression levels of CXCR4 mRNA was 7.17 ± 2.96 and 0.98 ± 0.06, respectively, showed that Calycosin can up-regulate the expression of miRNA-1246 in 769-P cell ( t=3.24, P<0.01), and down-regulate the expression of CXCR4 mRNA ( t=4.18, P<0.01). Compared with the control group, the Calycosin could down-regulate the expression of CXCR4 protein and ERK pathway protein in 769-P cell. Conclusion:Calycosin can inhibit the proliferation and migration of renal cancer 769-P cell, and its mechanism may be related to up-regulating the expression of miRNA-1246 and blocking the CXCR4/ERK pathway.

During orthodontic treatment,irregular and varying sized nodular bony protrusions may sometimes appear on the labial side of the patient's anterior alveolar bone,which is closely related to the differential bone remodeling on the inner and outer surfaces of the alveolar bone.Labial protuberances not only affect the aesthetic results of orthodontic treatment,but also pose potential risks to periodontal health.Currently,it is believed that the influencing factors of the formation of the labial protuberances may be related to the patient's gender and age,tooth movement speed,and extent of anterior teeth retraction.Labial protuberances typically resolve spontaneously,however,if persistent,alveoloplasty may be necessary for treatment.This review provides a summary on the occurrence,influencing factors of formation,potential biological mechanisms,and corresponding treatment methods of labial protuberances during orthodontic treatment.

Objective:To explore the relationship between the relative expression of miRNA-676-3p and the survival of renal cancer patients, and its effect on the proliferation and invasion of renal cancer by targeting and regulating prefoldin 1 (PFDN1).Methods:OncoRank online software was selected to analyze the relationship between the relative expression of miRNA-676-3p and the survival rate of renal cancer patients. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the relative expression of miRNA-676-3p in renal cancer cell lines. Renal carcinoma CAKI1 cells were resuscitated, and the transfected miRNA-NC was used as the control group, and the transfected precursor miRNA-676-3p was used as the overexpression group. The relative expression of miRNA-676-3p was detected by RT-qPCR. The cell absorbance and invasion number of the two groups were measured by CCK-8 and Transwell invasion assays, respectively. The target gene of miRNA-676-3p was predicted and verified by referring to the TargetScan Release 8.0 website and dual-luciferase reporter gene experiment. The expression of PFDN1 gene and Wnt/β-catenin molecular pathway protein in the two groups of cells were determined by RT-qPCR and Western blotting, respectively. Measurement data were expressed as mean ± standard deviation ( ± s), t-test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups. Results:The survival rate of renal cancer patients with high expression of miRNA-676-3p was significantly higher than that of renal cancer patients with low expression of miRNA-676-3p, the difference was statistically significant ( P<0.01). The relative expression of miRNA-676-3p in renal cancer cell lines was significantly lower than that in normal renal tubular epithelial cells, the difference was statistically significant ( P<0.01), and the relative expression of miRNA-676-3p in CAKI1 cells was the lowest, the difference was statistically significant ( P<0.01). The relative expression levels of miRNA-676-3p in the control and overexpression groups were 1.04±0.59 and 15.90±1.70, respectively, and the overexpression group was significantly higher than the control group, the difference was statistically significant ( P<0.01). After 24, 48, 60, and 72 h of culture, the absorbance of cells in the overexpression group was lower than that in the control group, the difference was statistically significant ( P<0.05). The number of invasion cells in the control group and the overexpression group were (115.90 ± 24.73) and (43.83 ± 21.94) cells, respectively, and the number of cell invasion in the overexpression group was significantly lower than that in the control group, the difference was statistically significant ( P<0.01). PFDN1 was the downstream target gene of miRNA-676-3p ( P<0.01). The relative expression of PFDN1 gene in the overexpression group was significantly lower than that in the control group, the difference was statistically significant ( P<0.01). The expression of Wnt/β-catenin molecular pathway proteins in the overexpression group was lower than that in the control group. Conclusions:Renal cancer patients with high expression of miRNA-676-3p had a higher survival rate. miRNA-676-3p inhibited the proliferation and invasion of renal cancer CAKI1 cells by significantly down-regulating the expression of PFDN1, thereby inhibiting the development of renal cancer.

Objective:To analyze the expression of long non-coding RNA (lncRNA) ZFPM2-AS1 in bladder cancer tissues and cell lines, and to observe the effect of down-regulating ZFPM2-AS1 on the migration and proliferation of bladder cancer cells and explore its molecular mechanism.Methods:Real-time quantitative fluorescent polymerase chain reaction (qRT-PCR) was used to detect the expression of ZFPM2-AS1 in 51 pairs of bladder cancer tissues and adjacent tissues, bladder cancer cell lines (J82, 5637, BIU-87, T24) and human normal bladder epithelial cells SV-HUC-1. The bladder cancer cells with the highest ZFPM2-AS1 expression were selected and transfected with the small interfering siRNA-ZFPM2-AS1 plasmid and the negative control plasmid, respectively, and defined as the experimental group and the control group. qRT-PCR was used to detect the expression of ZFPM2-AS1 in two groups of cells. Transwell migration test and tetramethylazozole blue (MTT) method were used to detect the cell migration ability and proliferation ability of the two groups. qRT-PCR was used to detect the expression of Up-frameshift mutant 1 (UPF1) mRNA in two groups of cells. Western blot was used to detect the expression of UPF1 and mTOR signaling pathway proteins in the two groups of cells.Results:The expression of ZFPM2-AS1 in bladder cancer tissues was significantly higher than that in adjacent tissues ( P<0.01). The expression of ZFPM2-AS1 in bladder cancer cell lines was significantly higher than that in human normal bladder epithelial cells ( P<0.01), and ZFPM2-AS1 had the highest expression in BIU-87 cells ( P<0.01). Compared with the control group, the expression of ZFPM2-AS1 in BIU-87 cells in the experimental group was significantly reduced [(1.01±0.06) vs (0.16±0.04), t=12.28, P<0.01]. Compared with the control group, the migration ability of BIU-87 cells in the experimental group was decreased ( P<0.05), and the proliferation ability of BIU-87 cells was significantly decreased from the second day ( P<0.05). Compared with the control group, UPF1 mRNA expression in BIU-87 cells in the experimental group was significantly decreased [(1.00±0.02) vs (0.28±0.04), t=15.49, P<0.01]. Western blot results showed that UPF1 protein expression and mammalian rapamycin target protein (mTOR), GRB2, IRS1 and p-PI3K signal pathway protein expression were decreased in BIU-87 cells. Conclusions:ZFPM2-AS1 is highly expressed in bladder cancer tissues and cell lines. Down-regulating ZFPM2-AS1 can inhibit the migration and proliferation of BIU-87 cells. The molecular mechanism may be related to the inhibition of UPF1 gene expression.

Objective:To observe the expression of microRNA (miRNA, miR) -7850 in renal cancer tissues, and to explore the effect of miR-7850 on the proliferation and migration of renal cancer cells and on the regulation of serine proteinase inhibitor B3 (SERPINB3) gene expression.Methods:Real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-7850 in renal cancer tissues and renal cancer cell lines. The renal cell carcinoma cell line with the lowest expression of miR-7850 was selected, and the negative control sequence (miR-NC) and miR-7850 mimics were transfected into renal cell carcinoma cells by Lipofectamine 2000 transfection reagent, respectively, which were defined as miR-NC group and miR-7850 group. qRT-PCR was used to detect the expression of miR-7850 in transfected renal cancer cells. The cell proliferation and migration ability after transfection were detected by cell counting kit-8 (CCK-8) method and transwell experiment. Bioinformatics prediction and dual luciferase reporter gene experiments were used to verify the target gene of miR-7850. qRT-PCR and Western blot were used to detect the expression of target genes in renal cancer cells after transfection.Results:Compared with adjacent tissues (5.95±0.44), the expression of miR-7850 in kidney cancer tissues (1.19±0.33) was lower ( P<0.01). Compared with immortalized proximal renal tubular epithelial cells (1.01±0.07), the expression of miR-7850 was lower in renal cancer cell lines ( P<0.05), and the lowest in A498 cells (0.13±0.01) ( P<0.01). The expression of miR-7850 in the miR-7850 group (7.46±0.93) was significantly higher than that in the miR-NC group (1.01±0.08) ( P<0.01), indicating successful transfection. Compared with the miR-NC group, the cell proliferation ability of the miR-7850 group was significantly reduced ( P<0.05). The number of migrating cells in miR-NC group and miR-7850 group were (139.50±12.31) and (75.09±16.05) cells, respectively, and the cell migration ability in miR-7850 group decreased significantly ( P<0.01). Bioinformatics technology shows that the target gene of miR-7850 was SERPINB3. The dual luciferase reporter gene experiment confirmed that miR-7850 can target the SERPINB3 gene ( P<0.05). Compared with the miR-NC group, the expression of SERPINB3 in cells of miR-7850 group was significantly reduced ( P<0.05), as well as the CDK4, CyclinD, Snail and Vimentin. Conclusions:miR-7850 is lowly expressed in renal cancer tissues and cell lines. miR-7850 can inhibit the proliferation and migration of renal cancer A498 cells, which may be related to its inhibition of SERPINB3 gene expression.