Objective:This study aims to investigate the alterations in m 6A methylation associated with age-related idiopathic pulmonary fibrosis(IPF). Methods:By collecting peripheral blood samples from IPF patients, we investigated the changes in m6A modification levels of total RNA and key regulatory factors in elderly IPF patients.Then, the pulmonary fibrosis models of young and old mice were constructed for verification.A total of 10 IPF patients and 10 healthy controls were selected for this study.The m 6A methylation quantitative kit was employed to assess the m 6A modification levels of total RNA.The expression levels of key m 6A methylation regulators, METTL3, METTL14, and FTO, were quantified using qRT-PCR.Additionally, thirty-two healthy male C57BL/6 mice, comprising 16 mice aged 10-12 weeks and 16 mice aged 6-7 months, were divided into four groups: young control(A), young pulmonary fibrosis(B), aged control(C), and aged pulmonary fibrosis(D), with 8 mice in each group.Mice in groups B and D were intratracheally administered bleomycin to establish a pulmonary fibrosis model, while those in groups A and C received normal saline.Twenty-eight days post-model establishment, the mice were euthanized, and lung tissues were collected for analysis.Histological evaluations were performed using hematoxylin and eosin(HE)staining, Masson staining, hydroxyproline content determination, and immunohistochemistry to assess the extent of pulmonary fibrosis.The m 6A methylation quantification kit was also utilized to measure the m 6A modification levels of total RNA in lung tissue.Furthermore, the mRNA and protein expression levels of the methyltransferase METTL3 were assessed by qRT-PCR and Western blot experiments. Results:The level of m 6A modification was significantly elevated in the aged IPF patient group(0.36±0.03)compared to the control group t=4.882( P<0.05).Furthermore, the expression of METTL3 was markedly higher in the aged IPF patients( t=6.082), while the expression of METTL14 was significantly lower t=17.58( P<0.05).In contrast, the expression level of FTO did not exhibit a significant difference.It is hypothesized that the increased m 6A modification of total RNA in aged IPF patients is closely associated with METTL3.Furthermore, the degree of lung fibrosis in aged mice was more severe than that in young mice.Immunohistochemistry results indicated that TGF-β1 expression was elevated in the lung fibrosis group, with higher levels observed in group D compared to group B( t=5.891, P<0.05), and in group C compared to group A t=4.135( P<0.05).The percentage of positive area for α-SMA was significantly greater in the lung fibrosis mouse model than in the control group t=20.08( P<0.05).The level of m 6A modification was increased in both lung fibrosis groups relative to the normal control group( P<0.05), although no significant difference was found between group D and group B. Overall, METTL3 mRNA and protein expression were upregulated in the lung fibrosis group, with expression in group D being lower than in group B( P<0.05). Conclusions:The level of m 6A modification is elevated in pulmonary fibrosis, and the expression of METTL3 is upregulated in this condition.The downregulation of METTL3 may be associated with the extent of aging, which subsequently exacerbates the progression of pulmonary fibrosis.

Objective:This study aims to investigate the alterations in m 6A methylation associated with age-related idiopathic pulmonary fibrosis(IPF). Methods:By collecting peripheral blood samples from IPF patients, we investigated the changes in m6A modification levels of total RNA and key regulatory factors in elderly IPF patients.Then, the pulmonary fibrosis models of young and old mice were constructed for verification.A total of 10 IPF patients and 10 healthy controls were selected for this study.The m 6A methylation quantitative kit was employed to assess the m 6A modification levels of total RNA.The expression levels of key m 6A methylation regulators, METTL3, METTL14, and FTO, were quantified using qRT-PCR.Additionally, thirty-two healthy male C57BL/6 mice, comprising 16 mice aged 10-12 weeks and 16 mice aged 6-7 months, were divided into four groups: young control(A), young pulmonary fibrosis(B), aged control(C), and aged pulmonary fibrosis(D), with 8 mice in each group.Mice in groups B and D were intratracheally administered bleomycin to establish a pulmonary fibrosis model, while those in groups A and C received normal saline.Twenty-eight days post-model establishment, the mice were euthanized, and lung tissues were collected for analysis.Histological evaluations were performed using hematoxylin and eosin(HE)staining, Masson staining, hydroxyproline content determination, and immunohistochemistry to assess the extent of pulmonary fibrosis.The m 6A methylation quantification kit was also utilized to measure the m 6A modification levels of total RNA in lung tissue.Furthermore, the mRNA and protein expression levels of the methyltransferase METTL3 were assessed by qRT-PCR and Western blot experiments. Results:The level of m 6A modification was significantly elevated in the aged IPF patient group(0.36±0.03)compared to the control group t=4.882( P<0.05).Furthermore, the expression of METTL3 was markedly higher in the aged IPF patients( t=6.082), while the expression of METTL14 was significantly lower t=17.58( P<0.05).In contrast, the expression level of FTO did not exhibit a significant difference.It is hypothesized that the increased m 6A modification of total RNA in aged IPF patients is closely associated with METTL3.Furthermore, the degree of lung fibrosis in aged mice was more severe than that in young mice.Immunohistochemistry results indicated that TGF-β1 expression was elevated in the lung fibrosis group, with higher levels observed in group D compared to group B( t=5.891, P<0.05), and in group C compared to group A t=4.135( P<0.05).The percentage of positive area for α-SMA was significantly greater in the lung fibrosis mouse model than in the control group t=20.08( P<0.05).The level of m 6A modification was increased in both lung fibrosis groups relative to the normal control group( P<0.05), although no significant difference was found between group D and group B. Overall, METTL3 mRNA and protein expression were upregulated in the lung fibrosis group, with expression in group D being lower than in group B( P<0.05). Conclusions:The level of m 6A modification is elevated in pulmonary fibrosis, and the expression of METTL3 is upregulated in this condition.The downregulation of METTL3 may be associated with the extent of aging, which subsequently exacerbates the progression of pulmonary fibrosis.

Objective:To investigate the correlation between immune function and age-related pulmonary fibrosis, as well as the potential impact of bacterial lysates on this condition.Methods:Twenty-four healthy male C57BL/6 mice, aged 24, were randomly divided into three groups: a control group(Group N), a pulmonary fibrosis group(Group M), and a pulmonary fibrosis+ bacterial lysis product intervention group(Group P). Mice in Groups M and P were intratracheally injected with bleomycin(5 mg/kg)to induce a mouse pulmonary fibrosis model, while mice in Group N were injected with saline.After modeling, mice in Group P were orally administered 0.4 ml of a bacterial lysis product once a day.After 28 days, lung tissue and blood samples were collected for analysis.Pathological changes in lung tissue were assessed using hematoxylin and tosin staining(HE)and Masson staining and the Ashcroft score.The expression of CD4+ and CD8+ in lung tissue was evaluated using immunohistochemistry.The levels of serum interferon-γ(INF-γ), interleukin-3(IL-13), and immunoglobulin A(IgA)protein were measured using Enzyme-linked Immuno Sorbent Assay(ELISA). The levels of INF-γ and IL-13 mRNA in lung tissue were determined using Real-Time Quantitative Transcription PCR(RT-qPCR). Additionally, the protein expression levels of matrix metalloprotein-9(MMP-9)and tissue inhibitor of metalloproteincise 1(TIMP-1)in lung tissue were assessed using blot analysis.Results:The degree of lung fibrosis was significantly reduced in mice in group P compared with group M when treated with bacterial lysis products.Group M showed a significant decrease in the expression of CD4+ T cells and an increase in the expression of CD8+ T cells( P<0.05)compared to group N. Additionally, the content of IgA was decreased( P<0.05)in group M. On the other hand, group P showed a significant increase in the expression of CD4+ T cells and a decrease in the expression of CD8+ T cells( P<0.05)compared to group M. Furthermore, the content of IgA was elevated( P<0.05)in group P. After bacterial lysis product intervention, mRNA and protein expression levels of IFN-γ were elevated( P<0.05), while mRNA and protein expression levels of IL-13 were reduced( P<0.05). Moreover, protein expression of MMP-9 and TIMP-1 was significantly up-regulated in group M compared with group N( P<0.05), and decreased after bacterial lysis product intervention( P<0.05). Conclusions:It is well-known that immune mechanisms play a crucial role in the development of pulmonary fibrosis.The use of bacterial lysates has been found to effectively regulate immune balance and mitigate the severity of pulmonary fibrosis in elderly mice.

The dental operative microscope has been widely employed in the field of dentistry, particularly in endodontics and operative dentistry, resulting in significant advancements in the effectiveness of root canal therapy, endodontic surgery, and dental restoration. However, the improper use of this microscope continues to be common in clinical settings, primarily due to operators' insufficient understanding and proficiency in both the features and established operating procedures of this equipment. In October 2019, Professor Jingping Liang, Vice Chairman of the Society of Cariology and Endodontology, Chinese Stomatological Association, organized a consensus meeting with Chinese experts in endodontics and operative dentistry. The objective of this meeting was to establish a standard operation procedure for the dental operative microscope. Subsequently, a consensus was reached and officially issued. Over the span of about four years, the content of this consensus has been further developed and improved through practical experience.
Humans ; Dentistry, Operative ; Consensus ; Endodontics ; Root Canal Therapy ; Dental Care

This paper summarizes the training experience of clinical skills competition in national medical colleges and universities, especially summarizes and analyzes the details of guiding competition. It's concluded that strengthening the basic knowledge and principal theory, and standardizing and mastering basic operation are the basis of training. The cultivation of clinical thinking ability and the mastery of the principles of diagnosis and treatment are the key points. And the teamwork and the psychological health of teachers and students are the essential conditions. National clinical skills competition has helped the medical students to reinforce their clinical practice ability and also made the clinical operation training system standardized, providing reference for clinical training of medical students in the future.

Objective:To investigate the effects of Spironolactone on expression of interleukin-33(IL-33)/ suluble tumorigenicity 2(ST2)in a rat model of diastolic heart failure.Methods:Wistar male rats were used to establish a model of diastolic heart failure, which was induced by abdominal aortic coarctation.Rats were randomly divided into the sham operation group, the model group and the Spironolactone group.Rats in the Spironolactone group were given Spironolactone 40 mg/kg/day by intragastric administration, and the sham operation and model groups were given the same amount of physiological saline by intragastric administration for 8 weeks.At the end of the experiment, end-diastolic interventricular wall thickness, left ventricular posterior wall end-diastolic thickness, left ventricular mass and mitral flow E/A were measured via echocardiography.Myocardial tissues were taken and the protein and mRNA expression of soluble ST2(sST2)and IL-33 in rat myocardium was detected by using real-time reverse transcription PCR(RT-PCR)and enzyme-linked immunosorbent assays(ELISA).Results:Compared with the sham operation group, echocardiography showed that end-diastolic interventricular thickness, left ventricular posterior wall end-diastolic thickness and left ventricular mass were increased and mitral flow E/A was decreased in the model group( P<0.05), indicating the model was successful.Compared with the model group, end-diastolic interventricular thickness(2.09±0.11) mm vs.(2.21±0.14) mm, end-diastolic left ventricular posterior wall thickness(2.08±0.17) mm vs.(2.36±0.14) mm and left ventricular mass(0.74±0.17) g vs.(1.35±0.22)g were decreased significantly and the mitral flow E/A ratio(1.44±0.25 vs.1.17±0.13)was elevated in the Spironolactone group( P<0.05), suggesting that spironolactone significantly improved left ventricular diastolic function.RT-PCR and ELISA results showed that sST2 mRNA and protein levels were lower and IL-33 mRNA and protein levels in myocardial tissues were higher in the spironolactone group than in the model group(0.16±0.01) vs.(0.56±0.07), (0.07±0.02)pg/ml vs.(0.35±0.21) pg/ml, (0.67±0.09) vs.0.23±0.05, (0.54±0.11) pg/ml vs.(0.31±0.03) pg/ml, P<0.05. Conclusions:Spironolactone can improve diastolic function in rats by decreasing the expression of sST2 mRNA and protein and enhancing the expression of IL-33 mRNA and protein.

Objective:To investigate the role and its mechanism of long non-coding RNA cell cyclin-dependent kinase inhibitor gene CDKN2B-AS1 and its adjacent gene CDKN2A in idiopathic pulmonary fibrosis coexisting with lung cancer.Methods:The cancerous lung tissue specimens and adjacent normal lung tissue specimens were collected from 8 patients with lung adenocarcinoma.The expression level of CDKN2B-AS1 and CDKN2A mRNA were determined by reverse transcription quantitative real-time PCR(RT-qPCR)assays.Pulmonary fibrosis cell model was established by treating human fetal lung fibroblast MRC-5 cell line that was selected as the study object.And the cells were randomly divided into 4 groups: the normal group, the intervention group[induced by transforming growth factou-β(TGF-β1)], the negative siRNA intervention group(induced by TGF-β1 and transfected by nonsense sequence siRNA)and the positive siRNA intervention group TGF-β1 and transfected by siRNA of CDKN2B-AS1). The morphological changes of each group were observed by using the inverted phase contrast microscope after 24 hours intervention.The expression levels of CDKN2B-AS1 and CDKN2A mRNA were determined by RT-qPCR, and the protein amounts of CDKN2A and P53 were measured by Western blotting.Results:The expression levels of CDKN2B-AS1 and CDKN2A mRNA were lower in lung cancer tissue than in adjacent normal lung tissues(2.60±1.33 vs.21.90±19.83, 0.34±0.10 vs.19.83±7.67, t=2.747 and 7.187, P<0.05), which were consistent with the results in IPF.In cell experiments, we observed that TGF-β1 intervention gradually transformed MRC-5 cells from multi-spindled or stellate structures to flattened fibroblasts with varying degrees of differentiation.Compared with the normal group, TGF-β1 intervention group showed that the expression levels of CDKN2B-AS1 and CDKN2A mRNA were significantly decreased(6.80±0.30 vs.56.12±2.46, 9.39±0.37 vs.64.54±3.89, t=47.746 and 33.797, both P<0.001). Compared with the intervention group, the expression levels of CDKN2B-AS1 and CDKN2A mRNA were further decreased in the positive siRNA intervention group(2.38±0.29 vs.6.80±0.30, 2.81±0.36 vs.9.39±0.37, t=4.279 and 4.032, P=0.003 and 0.004), and there was a positive correlation between the mRNA expressions of CDKN2B-AS1 and CDKN2A( r=0.988, P=0.000). Compared with the normal group, the intervention group showed that the protein expressions of CDKN2A and P53 were decreased(3.12±0.06 vs.4.12±0.59, 1.12±0.07 vs.2.11±0.06, t=2.921 and 19.599, P=0.043 and 0.000), and there was a positive correlation between the expression of CDKN2A and P53( r=0.772, P=0.000). Conclusions:Long non-coding RNA CDKN2B-AS1 has a low expression level in both idiopathic pulmonary fibrosis and lung cancer tissues, and it may participate in the P53 pathway by regulating the expression of the neighboring gene CDKN2A, which may be one reason for the high incidence of lung cancer in IPF patients.

Objective To explore the effect of age factor on the degree of pulmonary fibrosis and on the change in telomerase reverse transcriptase(TERT) activity in mice.Methods A total of 80 healthy male C57BL/6 mice aged 20-weeks were randomized into 4 groups:a young pulmonary fibrosis model group(n=20),a young control group(n=20),a senile pulmonary fibrosis model group(n=20),and an elderly control group (n =20).Two model groups were induced by an intratracheally injected bleomycin in 5 mg/kg,and two control groups by an intratracheally injected normal saline.The elderly were defined as 26 weeks old mice.Five mouse pulmonary fibrosis models and five mouse controls in four groups were randomly selected and killed at 7,14,21,28 days.Lung tissue specimens were stained with hematoxylin eosin (HE)and Masson trichrome (Masson)method,and then pathological changes were observed.In addition,immunohisto-chemical staining was used to observe the expression levels of epithelial cell marker protein E-cadherin(E-cad),stromal cell marker protein Vimentin,and alpha smooth muscle actin(α-SMA).Finally,the expression level of telomerase reverse transcriptase(TERT)protein was detected by Western blotting.Results A bleomycin-induced pulmonary fibrosis mice model was successfully prepared.The degree of pulmonary fibrosis was more severe in old mice than in young mice.Compared with the young pulmonary fibrosis model group,the expression level of E-Cad was decreased in the senile pulmonary fibrosis model group(P ＜0.05).Compared with the young control group and the elderly control group,the expression levels of E-Cad in the young pulmonary fibrosis model group and the senile pulmonary fibrosis model group were decreased (P ＜ 0.05),and decreased along with the prolongation of the modeling time.Compared with the young pulmonary fibrosis model group,the expression levels of alpha-SMA and vimentin were increased in the senile pulmonary fibrosis model group(P ＜ 0.05).The expression levels of alpha-SMA and vimentin were increased in the young pulmonary fibrosis model group and the senile pulmonary fibrosis model group,as compared to the young control group and the elderly control group(P＜0.05),and increased along with the prolongation of the modeling time.The activity of TERT in lung tissue of mice was increased at first and then decreased.Compared with the young pulmonary fibrosis model group,the activity of TERT in the senile pulmonary fibrosis model group significantly fluctuated(P＜0.05).Conclusions Age factor can affect the severity of pulmonary fibrosis by affecting the activity of telomerase reverse transcriptase in mouse models of pulmonary fibrosis.

Objective To investigate the effect of protein kinase CK2α expression on the transformation of epithelium in human laryngeal carcinoma Hep-2 cells.Methods HepG 2 cells were transfected into human laryn-geal squamous cell carcinoma cell line Hep-2 by immunization with pGCsi-H1-CK2α in vitro and divided into un-transfected group(pGCsi-H1-control)and interfering plasmid(PGCsi-H1-CK2α).The cells were transfected with G418 for 24 h and identified by Real-time PCR and Western blot.The invasion and migration ability of Hep-2 cells were detected by Transwell chamber.The patients were observed by phase contrast microscope the expression of E-cadherin,vimentin and transcription factors slug and snail were detected by Western blot. Results The expression of CK2α gene in the transfected group was significantly lower than that in the untransfected group and control plas-mid(P<0.01).Transwell experiments showed that stable knockout of CK2α prevented the migration and invasion of Hep-2 cells.Compared with untransfected and control plasmid transfection groups,the expression of E-cadherin in the interfering plasmid transfection group increased,but the expression of snail,slug and vimentin decreased. Conclusions RNAi mediated inhibition of CK2α inhibits the transformation of epithelial mesenchymal cells in la-ryngeal squamous cell carcinoma.

Objective To apply the Chinese version of Care Partner-Frailty Index-Comprehensive Geriatric Assessment (CP-FI-CGA) in Taiyuan part of elderly patients and study theirs frailty conditions. Methods To Choose the Chinese version of CP-FI-CGA questionnaire, patients′ general information questionnaire and Clinical Frailty Scale to evaluate 385 patients and analyze the results statistically. Results Of 385 patients, female patients were 166 cases (43.12%);the frailty index score was (0.318 ± 0.165) points, the CFS was 5.044 ± 1.483. Single factor analysis showed that age, marital status, the kinds of medication which the patients used, how much help the patient required, the condition of social support, and the sleep state, these six factors had statistical significance (Z=-7.292, Z=-1.994, χ2=27.726, Z=-9.688,χ2=8.117,χ2=53.477, all P<0.01). Multiple linear regression analysis showed that age, the kinds of medication which the patients used, how much help the patient required and the sleep state, these four factors were independent factors (model R=0.610, R2=0.372; adjusted R2=0.362, F=37.241, P< 0.01). Conclusions CP-FI-CGA questionnaire can accurately estimate the frailty degree by evaluating patients′each system and can be promoted in the clinical geriatric ward and nursing home, etc.