BACKGROUND:Human periodontal stem cells have a certain inhibitory effect on the pro-inflammatory function of M1-type macrophages,and it is not clear whether icariin,which has anti-inflammatory and other pharmacological activities,can enhance the inhibitory effect of human periodontal stem cells on M1-type macrophages. OBJECTIVE:To investigate the effect of icariin on M1 macrophages after pretreatment of human periodontal stem cells. METHODS:Primary human periodontal stem cells were isolated,cultured and characterized.THP-1 was induced and M1-type macrophages were identified by immunofluorescence staining and PCR.Human periodontal stem cells were cultured with α-MEM complete medium containing concentrations of 10-7,10-6,10-5,and 10-4 mol/L icariin,and the cytotoxicity of Icariin on human periodontal stem cells was detected by the CCK-8 assay at 1,3,5,and 7 days,respectively.α-MEM complete medium,untreated α-MEM conditioned medium for human periodontal stem cells and α-MEM conditioned medium for human periodontal stem cells pretreated with icariin for 24 hours were conditioned with RPMI-1640 complete medium in a 1:1 ratio for M1-type macrophages in the control,untreated,and pretreated groups,and 24 hours later,the mRNA expression of inflammatory factors in M1 macrophages was detected by RT-PCR.The protein expression of inflammatory factors in M1 macrophages was detected by ELISA.The expression of surface markers and nuclear factor-κB pathway-related proteins in M1/M2 macrophages was detected by western blot assay. RESULTS AND CONCLUSION:(1)CCK-8 assay results showed that 10-7,10-6,10-5,10-4 mol/L icariin was not cytotoxic to the human periodontal stem cells,and from day 5 onwards,all the concentrations increased the cell viability,and promoted the cell proliferation.10-4 mol/L icariin was selected for follow-up experiment.(2)RT-PCR and ELISA results showed that compared with the control group,the untreated group and the pretreated group both decreased the expression and secretion of interleukin-1β,interleukin-6,and tumor necrosis factor-α of M1-type macrophages(P<0.05),and the pretreated group was lower than the untreated group(P<0.05).(3)Western blot assay results showed that compared with the untreated group,the expression of CD86 was significantly lower in the pretreated group(P<0.05);compared with the control group,the expression of CD206,a surface marker of M2-type macrophages,was elevated in both the untreated and pretreated groups(P<0.01),and it was significantly higher in the pretreated group than in the untreated group(P<0.01).In M1-type macrophages after 24 hours of conditioned culture,compared with the control group,the expression of nuclear factor-κB/P65 was decreased in the untreated group and the pretreated group(P<0.01),and the expression of p-IκBα was decreased only in the pretreated group(P<0.01);the expression of both nuclear factor-κB/P65 and p-IκBα was significantly reduced in the pretreated group compared with the untreated group(P<0.05),while the difference of IκBα in the three groups was not statistically significant.(4)These results indicated that icariin enhanced the inhibitory effect of human periodontal stem cells on M1-type macrophages,and this effect may be related to the inhibition of the nuclear factor-κB signaling pathway of macrophages.

ObjectiveTo clarify the protective effect of Danlou tablet， a representative traditional Chinese medicine of the stagnation of phlegm and blood stasis co-treatment method， on vascular endothelial function in patients with atherosclerosis （AS）. MethodsA randomized controlled trial was conducted. From September 2023 to November 2023， a total of 72 patients who were diagnosed at Guang'anmen Hospital， China Academy of Chinese Medical Sciences and met the inclusion and exclusion criteria for carotid atherosclerosis （CAS） combined with coronary atherosclerotic heart disease （CHD） and stable angina pectoris （SAP） were enrolled. The patients were randomly divided into a control group （receiving conventional Western medicine treatment） and an observation group （receiving Danlou tablet combined with conventional Western medicine treatment）， with 36 cases in each group. The intervention lasted for 12 weeks. The frequency of angina pectoris attacks was recorded to evaluate the clinical efficacy of Danlou tablet. Peripheral blood samples were collected from patients， and the expression levels of serum endothelial injury markers before and after treatment were detected by enzyme-linked immunosorbent assay （ELISA）. The nitrate reductase method was employed to evaluate the protective effect of Danlou tablet on vascular function. The expression levels of serum inflammatory factors and lipoproteins were determined by ELISA and an automatic biochemical analyzer （dynamic timed scatter turbidimetry and enzymatic method） to assess the anti-inflammatory and lipid-regulating effects of Danlou tablet. ResultsIn terms of angina pectoris attacks， compared with that in the control group， the frequency of attacks in the observation group was reduced （P<0.05）. In terms of endothelial injury markers， compared with the levels before treatment within the same group， the levels of endothelin-1 （ET-1）， intercellular adhesion molecule-1 （ICAM-1）， and vascular cell adhesion molecule-1 （VCAM-1） in the peripheral blood of the observation group were decreased （P<0.05）， while the levels of nitric oxide （NO） and vascular endothelial growth factor （VEGF） were increased （P<0.05）. Compared with the control group after treatment， the differences in ET-1， NO， ICAM-1， and VCAM-1 were significant （P<0.05）. In terms of serum inflammatory factors， after treatment， the interleukin-6 （IL-6） level in the observation group was decreased significantly （P<0.05）. Compared with the control group after treatment， the IL-6 level in the observation group was decreased significantly （P<0.01）. In terms of serum lipoproteins， after treatment， the level of low-density lipoprotein cholesterol （LDL-C） in the observation group was decreased （P<0.05）. After treatment， the level of high-density lipoprotein cholesterol （HDL-C） in the observation group was significantly higher than that in the control group （P<0.05）. In terms of safety evaluation， no serious adverse events occurred in either group during the intervention period. ConclusionDanlou tablet applied to patients with CAS combined with CHD can improve endothelial function， reduce inflammatory indicators， alleviate symptoms， improve the quality of life of patients， and demonstrate good safety.

ObjectiveTo clarify the protective effect of Danlou tablet， a representative traditional Chinese medicine of the stagnation of phlegm and blood stasis co-treatment method， on vascular endothelial function in patients with atherosclerosis （AS）. MethodsA randomized controlled trial was conducted. From September 2023 to November 2023， a total of 72 patients who were diagnosed at Guang'anmen Hospital， China Academy of Chinese Medical Sciences and met the inclusion and exclusion criteria for carotid atherosclerosis （CAS） combined with coronary atherosclerotic heart disease （CHD） and stable angina pectoris （SAP） were enrolled. The patients were randomly divided into a control group （receiving conventional Western medicine treatment） and an observation group （receiving Danlou tablet combined with conventional Western medicine treatment）， with 36 cases in each group. The intervention lasted for 12 weeks. The frequency of angina pectoris attacks was recorded to evaluate the clinical efficacy of Danlou tablet. Peripheral blood samples were collected from patients， and the expression levels of serum endothelial injury markers before and after treatment were detected by enzyme-linked immunosorbent assay （ELISA）. The nitrate reductase method was employed to evaluate the protective effect of Danlou tablet on vascular function. The expression levels of serum inflammatory factors and lipoproteins were determined by ELISA and an automatic biochemical analyzer （dynamic timed scatter turbidimetry and enzymatic method） to assess the anti-inflammatory and lipid-regulating effects of Danlou tablet. ResultsIn terms of angina pectoris attacks， compared with that in the control group， the frequency of attacks in the observation group was reduced （P<0.05）. In terms of endothelial injury markers， compared with the levels before treatment within the same group， the levels of endothelin-1 （ET-1）， intercellular adhesion molecule-1 （ICAM-1）， and vascular cell adhesion molecule-1 （VCAM-1） in the peripheral blood of the observation group were decreased （P<0.05）， while the levels of nitric oxide （NO） and vascular endothelial growth factor （VEGF） were increased （P<0.05）. Compared with the control group after treatment， the differences in ET-1， NO， ICAM-1， and VCAM-1 were significant （P<0.05）. In terms of serum inflammatory factors， after treatment， the interleukin-6 （IL-6） level in the observation group was decreased significantly （P<0.05）. Compared with the control group after treatment， the IL-6 level in the observation group was decreased significantly （P<0.01）. In terms of serum lipoproteins， after treatment， the level of low-density lipoprotein cholesterol （LDL-C） in the observation group was decreased （P<0.05）. After treatment， the level of high-density lipoprotein cholesterol （HDL-C） in the observation group was significantly higher than that in the control group （P<0.05）. In terms of safety evaluation， no serious adverse events occurred in either group during the intervention period. ConclusionDanlou tablet applied to patients with CAS combined with CHD can improve endothelial function， reduce inflammatory indicators， alleviate symptoms， improve the quality of life of patients， and demonstrate good safety.

Purpose: This study aimed to investigate the molecular characteristics associated with better prognosis in breast cancer. Methods: We performed targeted sequencing of 962 genes in 56 samples, categorizing them into long-term and short-term survival groups as well as chemotherapy-sensitive and chemotherapy-resistant groups for further analyses. Results: The results indicated that the tumor mutational burden values were significantly higher in the short-term survival and chemotherapy-resistant groups (p = 0.008 and p = 0.003, respectively). Somatic mutation analysis revealed that the mutation frequencies of BCL9L and WHSC1 were significantly lower in the long-term survival group than those in the short-term survival group (p = 0.029 and p = 0.024, respectively). CREB-regulated transcription coactivator 1 (CRTC1) mutations occurred significantly more frequently in the chemotherapy-resistant group (p = 0.027) and were associated with shorter progression-free survival (p = 0.036).Signature weighting analysis showed a significant increase in Signature.3, which is associated with homologous recombination repair deficiency in the chemotherapy-sensitive group (p = 0.045). Conversely, signatures related to effective DNA repair mechanisms, Signature.1 and Signature.15, were significantly reduced (p = 0.002 and p < 0.001, respectively). Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated that gene mutations were significantly enriched in the JAK-STAT signaling pathway. Conclusion This study, through intergroup comparative analysis, found that immunotherapy (using programmed death 1/programmed death-ligand 1 inhibitors) may improve the prognosis of patients with short survival and chemotherapy resistance. Additionally, the study revealed that mutations in BCL9L and WHSC1 could serve as biomarkers for breast cancer prognosis, while CRTC1 mutations and Signature.3 could predict chemotherapy response. The study also found that the JAK-STAT pathway might be a potential therapeutic target for chemotherapy resistance. Therefore, this study identifies molecular characteristics that influence the prognosis of breast cancer patients, providing important theoretical insights for the development of personalized treatment strategies.

Purpose: This study aimed to investigate the molecular characteristics associated with better prognosis in breast cancer. Methods: We performed targeted sequencing of 962 genes in 56 samples, categorizing them into long-term and short-term survival groups as well as chemotherapy-sensitive and chemotherapy-resistant groups for further analyses. Results: The results indicated that the tumor mutational burden values were significantly higher in the short-term survival and chemotherapy-resistant groups (p = 0.008 and p = 0.003, respectively). Somatic mutation analysis revealed that the mutation frequencies of BCL9L and WHSC1 were significantly lower in the long-term survival group than those in the short-term survival group (p = 0.029 and p = 0.024, respectively). CREB-regulated transcription coactivator 1 (CRTC1) mutations occurred significantly more frequently in the chemotherapy-resistant group (p = 0.027) and were associated with shorter progression-free survival (p = 0.036).Signature weighting analysis showed a significant increase in Signature.3, which is associated with homologous recombination repair deficiency in the chemotherapy-sensitive group (p = 0.045). Conversely, signatures related to effective DNA repair mechanisms, Signature.1 and Signature.15, were significantly reduced (p = 0.002 and p < 0.001, respectively). Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated that gene mutations were significantly enriched in the JAK-STAT signaling pathway. Conclusion This study, through intergroup comparative analysis, found that immunotherapy (using programmed death 1/programmed death-ligand 1 inhibitors) may improve the prognosis of patients with short survival and chemotherapy resistance. Additionally, the study revealed that mutations in BCL9L and WHSC1 could serve as biomarkers for breast cancer prognosis, while CRTC1 mutations and Signature.3 could predict chemotherapy response. The study also found that the JAK-STAT pathway might be a potential therapeutic target for chemotherapy resistance. Therefore, this study identifies molecular characteristics that influence the prognosis of breast cancer patients, providing important theoretical insights for the development of personalized treatment strategies.

Purpose: This study aimed to investigate the molecular characteristics associated with better prognosis in breast cancer. Methods: We performed targeted sequencing of 962 genes in 56 samples, categorizing them into long-term and short-term survival groups as well as chemotherapy-sensitive and chemotherapy-resistant groups for further analyses. Results: The results indicated that the tumor mutational burden values were significantly higher in the short-term survival and chemotherapy-resistant groups (p = 0.008 and p = 0.003, respectively). Somatic mutation analysis revealed that the mutation frequencies of BCL9L and WHSC1 were significantly lower in the long-term survival group than those in the short-term survival group (p = 0.029 and p = 0.024, respectively). CREB-regulated transcription coactivator 1 (CRTC1) mutations occurred significantly more frequently in the chemotherapy-resistant group (p = 0.027) and were associated with shorter progression-free survival (p = 0.036).Signature weighting analysis showed a significant increase in Signature.3, which is associated with homologous recombination repair deficiency in the chemotherapy-sensitive group (p = 0.045). Conversely, signatures related to effective DNA repair mechanisms, Signature.1 and Signature.15, were significantly reduced (p = 0.002 and p < 0.001, respectively). Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated that gene mutations were significantly enriched in the JAK-STAT signaling pathway. Conclusion This study, through intergroup comparative analysis, found that immunotherapy (using programmed death 1/programmed death-ligand 1 inhibitors) may improve the prognosis of patients with short survival and chemotherapy resistance. Additionally, the study revealed that mutations in BCL9L and WHSC1 could serve as biomarkers for breast cancer prognosis, while CRTC1 mutations and Signature.3 could predict chemotherapy response. The study also found that the JAK-STAT pathway might be a potential therapeutic target for chemotherapy resistance. Therefore, this study identifies molecular characteristics that influence the prognosis of breast cancer patients, providing important theoretical insights for the development of personalized treatment strategies.

Objective To investigate the therapeutic effects and mechanisms of Maxing Shigan Decoction on cough variant asthma(CVA)rats.Methods Sixty rats were randomly divided into normal group,model group,low and high dose groups of Maxing Shigan Decoction,and high-dose of Maxing Shigan Decoction + signal transducer and activator of transcription 3(STAT3)activator Colivelin(Col)group,12 rats in each group.Except for the normal group,the CVA model was constructed by intraperitoneal injection of ovalbumin combined with moxa fumigation in all other groups of rats.After the corresponding treatment,the rats were observed for signs and cough counts,airway resistance(RE)was detected by pulmonary function meter,eosinophils(EOS)were counted by Diff-Quik staining,histopathological features of the lungs and bronchial tubes were observed by hematoxylin-eosin(HE)staining method,and the lung tissues were detected by enzyme-linked immunosorbent assay(ELISA)for monocyte chemotactic protein 1(MCP-1),and tumor necrosis factor α(TNF-α),and the protein expression levels of interleukin 6(IL-6),STAT3,and transient receptor potential vanilloid-1 channel(TRPV1)were detected by Western Blot.Results Compared with the normal group,rats in the model group showed obvious asthma symptoms,severe inflammatory cell infiltration was seen in the lung tissue,bronchial epithelial cell necrosis,ciliated adhesion,mucus,and RE,EOS number,MCP-1 and TNF-α contents,and protein expression levels of IL-6,STAT3,TRPV1 were elevated(P＜0.05);compared with the model group,rats in the low-and high-dose groups of Maxing Shigan Decoction showed significant improvement in asthma symptoms,reduction in lung and bronchial injury,and dose-dependent reduction in RE,EOS number,MCP-1 and TNF-α contents,and protein expression levels of IL-6,STAT3,and TRPV1(P＜0.05);compared with the high-dose group of Maxing Shigan Decoction,the rats in the high-dose Maxing Shigan Decoction+Col group showed increased asthma,increased lung and bronchial injury,and increased RE,EOS number,MCP-1 and TNF-α contents,and protein expression levels of IL-6,STAT3,and TRPV1(P＜0.05).Conclusion Maxing Shigan Decoction can effectively improve cough variant asthma in rats,and its mechanism is related to the inhibition of IL-6/STAT3 signaling pathway and the high expression of TRPV1.

BACKGROUND:Intervertebral disc degeneration is the basis of spinal degenerative diseases;however,there is no effective treatment. OBJECTIVE:To investigate whether sinomenine can inhibit interleukin-1β-induced apoptosis in nucleus pulposus cells and its molecular mechanism. METHODS:Rat nucleus pulposus cells were cultured in vitro by trypsin combined with type II collagenase digestion,and the cell growth curve was plotted.An appropriate sinomenine concentration was determined using the cell counting kit-8 kit.Nucleus pulposus cells were divided into control group,sinomenine group,interleukin-1β group,sinomenine+interleukin-1β group,zinc protoporphyrin group,zinc protoporphyrin+sinomenine group,zinc protoporphyrin+interleukin-1β group,and sinomenine+zinc protoporphyrin+interleukin-1β group.Proliferative activity,reactive oxygen species content,apoptosis rate,and heme oxygenase-1 expression in nucleus pulposus cells were detected. RESULTS AND CONCLUSION:The rat nucleus pulposus cells cultured in vitro were polygonal,triangular,and short wedge-shaped,and the cell growth showed an"S"curve.The cells grew slowly in the first 3 days of culture,rapidly in 4-6 days,and slowly again in 7-8 days.The cells then entered the"platform stage"where the number of cells no longer increased.The proliferative activity of myeloid cells showed no significant changes when the concentration of sinomenine was≤80 μmol/L(P>0.05).Interleukin-1β significantly reduced the proliferative activity of nucleus pulposus cells,increased the content of reactive oxygen species and led to apoptosis(P<0.01).Sinomenine intervention not only promoted heme oxygenase-1 expression(P<0.05)but also inhibited interleukin-1β-induced decrease in proliferative activity and increase in reactive oxygen species content and apoptosis rate in nucleus pulposus cells(P<0.05).These effects could be reversed by zinc protoporphyrin(P<0.01).

BACKGROUND:The application of miniscrew in adolescents is increasing day by day,but at present,there are few studies on bone mass in the external oblique line of the mandible in adolescents at home and abroad,and there is no systematic study on bone mass in the external oblique line of the mandible in adolescents in different growth and development periods. OBJECTIVE:To measure the bone mass in the external oblique line of the mandible in adolescents with different cervical vertebral bone ages using a cone-beam CT and to investigate the difference of bone mass in the external oblique line of the mandible in adolescents with different cervical vertebral bone ages and the correlation between bone mass in this area and the cervical vertebral bone age. METHODS:The cone-beam CT data of 105 adolescent patients before orthodontic treatment were collected and divided into CS3 group(n=24),CS4 group(n=26),CS5 group(n=29)and CS6 group(n=26)using the cervical vertebral maturation method.The adolescent mandibular buccal shelf was reconstructed by Mimics Medical 21.0 software.The width of buccal bone at 6 and 11 mm under the cemento-enamel junction and the bone height at 4 and 5 mm buccal to the cemento-enamel junction of right mandibular first and second molars were measured.The measured data were statistically analyzed.The measurement was made on four planes:plane 1 is the plane where the proximal mesial root of the mandibular right first molar is located;plane 2 is the plane where the distal mesial root of the mandibular right first molar is located;plane 3 is the plane where the proximal mesial root of the mandibular right second molar is located;and plane 4 is the plane where the distal mesial root of the mandibular right second molar is located. RESULTS AND CONCLUSION:In each group,the bone width on the buccal side of the external oblique line increased gradually from the first molar proximally to the second molar distally in adolescents,and the width of buccal bone at 6 and 11 mm under the cemento-enamel junction showed significant difference among different layers(P<0.05).The bone width of buccal bone at 11 mm under the cemento-enamel junction was greater than that at 6 mm.The bone height on the buccal side of the external oblique line increased gradually from the first molar proximally to the second molar distally in all four groups,and the bone height at 4 and 5 mm buccal to the cemento-enamel junction showed significant differences at different layers(P<0.05).The bone height at 4 mm buccal to the cemento-enamel junction was greater than that at 5 mm.On the fourth plane,the bone width at 11 mm buccal to the cemento-enamel junction was smaller in the CS3,CS4,and CS5 groups than in the CS6 group(P<0.05).On the third plane,the bone heights at 4 mm and 5 mm buccal to the cemento-enamel junction were smaller in the CS3 and CS4 groups than in the CS6 group(P<0.05).On the fourth plane,the bone height at 5 mm buccal to the cemento-enamel junction was smaller in the CS3 and CS4 groups than in the CS6 group(P<0.05).On the fourth plane,the bone height at 4 mm buccal to the cemento-enamel junction was smaller in the CS3 group than in the CS6 group(P<0.05).Spearman correlation analysis showed that there was no correlation between bone mass and the cervical vertebral bone age,except that there was a weak correlation between bone mass at some measurement sites and cervical vertebral bone age.To conclude,the bone mass in the external oblique area of the mandible in adolescents does not change significantly with the increase of cervical vertebral bone age.The buccal side of the mesial root and distal root of the mandibular second molar in the external oblique area of CS3-CS6 adolescents meets the requirement of bone mass for miniscrew implantation,which is a site available for miniscrew implantation.

BACKGROUND:Intervertebral disc degeneration is caused by damage and degeneration of the nucleus pulposus and annulus fibrosus tissues inside the intervertebral disc,resulting in structural and functional changes of the intervertebral disc.However,there is yet no effective drug treatment for intervertebral disc degeneration. OBJECTIVE:To investigate the inhibitory effect of syringin on intervertebral disc degeneration. METHODS:A total of 10 male Sprague-Dawley rats were selected,and the coccygeal intervertebral disc(Co4/Co5)of each rat was set as model group,Co5/Co6 intervertebral disc as syringin group,and Co6/Co7 intervertebral disc as control group.The control group did not receive any treatment.In the model group and syringin group,a miniature puncture needle was used to puncture the annulus fibrosus to establish an intervertebral disc degeneration model.Immediately after modeling,2.5 μL of normal saline and syringin solution(5 μmol/L)were given in the model and syringin groups,respectively.Four weeks after injection,the samples were taken.The degree of intervertebral disc degeneration in rats was observed by hematoxylin-eosin and safranine O-fast green staining.The expressions of type Ⅱ collagen,aggrecan and matrix metalloproteinases 3 and 13 in intervertebral disc tissue were analyzed by immunohistochemical staining. RESULTS AND CONCLUSION:Hematoxylin-eosin staining showed that in the model group,the height of intervertebral disc decreased,the cartilage endplate became thinner and cracked,the fibrous ring structure was disordered and cracked,and the nucleus pulposus disappeared;in the syringin group,the height of intervertebral disc was normal or slightly lower than that in the control group,the degree of cartilage endplate degeneration was lighter than that in the model group,the fiber circle permutation was relatively regular with no cracks,and the nucleus pulposus was partially shrunk.Safranine O-fast green staining showed that in the model group,the cartilage endplate of the intervertebral disc was defective and the calcified layer of cartilage became thinner,showing obvious degeneration.The structure and morphology of intervertebral disc cartilage endplate in the syringin group recovered to some extent.Immunohistochemical staining showed that,compared with the control group,the expressions of type Ⅱ collagen and aggrecan in the intervertebral disc cartilage were decreased in the model group(P<0.000 1),while the expressions of matrix metalloproteinases 3 and 13 increased(P<0.000 1).Compared with the model group,the expressions of type Ⅱ collagen and aggrecan in the intervertebral disc cartilage tissue were increased in the syringin group(P<0.001,P<0.000 1),while the expressions of matrix metalloproteinases 3 and 13 decreased(P<0.001,P<0.000 1).These results showed that syringin could improve the structure and function of intervertebral disc by inhibiting the expression of matrix metalloproteinases 3 and 13 and increasing the expression of type Ⅱ collagen and aggrecan,thus preventing and slowing down the procession of intervertebral disc degeneration.