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

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.

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 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.

Objective To investigate the role of Furin in breast cancer cell proliferation and provide a theoretical basis for in￣depth study of breast cancer. Methods Different concentrations of Furin inhibitor were added in MCF￣7 cell culture to test MCF￣7 cell proliferation by MTT essay.Hochest 33342 staining was used to detect the morphological change of apoptosic cells.Western blot analysis was applied to measure the level of cell apoptosis associated proteins,such as Caspase￣3,Caspase￣8 andCaspase￣9.The enzyme￣linked immunosorbent assay was used for detection the CAT and SOD levels in cell culture. ResultsMCF￣7 cell growth was inhibited by Furin inhibitor in a time and dose dependent manner.The results of Western blot and Hochest33342 staining indicated that MCF￣7 cells were apoptosis after Furin inhibitor treatment. The level of CAT was increasedsignificantly,associated with the level of SOD. Conclusion Furin inhibitor could induce MCF cell apoptosis, thereby inhibitcell proliferation by modulating MCF￣7 cell redox state.

Objective To observe the activation of anti-viral innate immune response of type Ⅰinterferon and inhibition of hepatitis B virus (HBV)genome replication in mice by HBV-3p-siRNA. Methods HBV-3p-siRNA was designed by targeting specific sequence of HBV S/P mRNA and was generated by in vitro transcription.Negative control siRNA (NC-siRNA)and non-modified HBV-siRNA were used as control groups.Blood samples were collected from tail vein of mice and the model of HBV-infected mice were established by hydrodynamic injection.Forty mice were divided into 4 groups with 10 in each group.The model group was only injected with pGL3.0-HBV1 .2 copy plasmid.The negative control group received peritoneal injection of NC-siRNA.HBV-siRNA group received peritoneal injection of HBV-siRNA and HBV-3p-siRNA group received peritoneal injection of HBV-3p-siRNA.The interferon-β(IFN-β)and hepatitis B surface antigen (HBsAg)in serum were detected by enzyme linked immunosorbent assay (ELISA).The copies of HBV DNA were assessed by fluore scence quantitative polymerase chain reaction (PCR ).The statistical difference between groups was determined using One way-ANOVA analysis by LSD or Dunnett T3.Results Serum level of IFN-β was (12.37±5 .32)pg/mL in model group,(22.61 ±6.29 )pg/mL in negative control group,(26.40±5 .39)pg/mL in HBV-siRNA group and (68.37± 21 .00 ) pg/mL in HBV-3p-siRNA group.The secretions of IFN-β into serum were significantly enhanced by HBV-siRNA and HBV-3p-siRNA compared with model group (F =23.988 and 46.523,respectively,both P <0.01).Serum level of HBsAg was (2 864.86±907.11 )ng/mL in model group,(2 198.86±456.89 )ng/mL in negative control group,(1 049.71 ± 396.28 )ng/mL in HBV-siRNA group and (640.86±383.08)ng/mL in HBV-3p-siRNA group.The expressions of HBsAg were inhibited by HBV-3p-siRNA and HBV-siRNA compared with model group (F = 23.537 and 39.144, respectively;P =0.025 and 0.010,respectively).Serum level of HBV DNA was (2.54 ×104 ±1 .46 × 104 )copy/mL in model group,(2.22×104 ±2.62×103 )copy/mL in negative control group,(3.59×103 ±2.88×103 )copy/mL in HBV-siRNA group and (2.65 ×103 ±1 .46×103 )copy/mL in HBV-3p-siRNA group.Serum level of HBV DNA were inhibited by HBV-3p-siRNA and HBV-siRNA compared with model group (F =15 .013 and 16.741 ,respectively,both P <0.05 ).All of the indicated siRNA used in the experiments showed no apparent effects on the body mass index of the mice models.Conclusion HBV-3p-siRNA,which induces the production of IFN-β and inhibits HBV replication through gene silencing in vivo ,may be a powerful bifunctional antiviral molecule.