Objective: This study aims to explore the association between oral lichen planus (OLP) and Hashimoto’s thyroiditis (HT) and its anti-thyroid antibodies to provide clinical evidence for thyroid disease screening in patients with OLP. Methods: This study was approved by the institutional ethics committee. A total of 125 clinically and histopathologically confirmed patients with OLP were enrolled as the case group, and they were matched with 125 non-OLP controls based on sex and age. Demographic data (gender, age, lesion type, and disease duration) were collected from both groups. Serum levels of thyroid peroxidase antibodies (TPOAb) and thyroglobulin antibodies (TgAb) were measured to analyze their associations with sex, age, lesion type, and disease duration in patients with OLP. Result: The prevalence of HT in patients with OLP was 31.20%, significantly higher than that in the control group (9.60%) (χ2=18.504, P<0.001). The prevalence of HT in female patients with OLP (39.13%) was significantly higher than that in male patients (9.09%)（χ2=10.93，P<0.001）. The positivity rate of thyroid peroxidase antibodies (TPOAb) in patients with OLP (17.6%) was significantly higher than in the control group (4.0%) (χ2=10.989, P<0.001). The TPOAb positivity rate was significantly higher in female patients (22.83%) than in male patients (3.03%) (χ2=5.210, P=0.014). There was no statistically significant difference in the positivity rate of TgAb between patients with OLP (7.2%) and the control group (3.2%) (P>0.05). Patients with erosive lesions had a significantly higher TPOAb positivity rate (25.0%, 17/68) compared to those with non-erosive lesions (8.77%, 5/57), and the difference was statistically significant (χ2=4.831, P=0.028). Logistic regression analysis revealed that female patients with OLP had an 8.935-fold higher risk of being TPOAb positive compared to males (OR=8.935, 95%CI: 1.134-70.388, P=0.038). Patients with erosive OLP lesions had a 3.199-fold higher risk of TPOAb positivity compared to those with non-erosive lesions (OR=3.199, 95%CI: 1.064-9.618, P=0.038). Conclusion The prevalence of HT is higher in patients with OLP, with higher positivity rates of anti-thyroid antibodies observed in female patients and those with erosive OLP lesions. This suggests that thyroid disease screening should be incorporated into the clinical management of patients with OLP, especially for women and patients who present with erosive lesions.

Objective To construct a universal influenza mRNA vaccine and evaluate its immunogenicity.Methods The antigen sequence of hemagglutinin(HA),nucleoprotein(NP)and matrix protein 2 ectodomain(M2e)in influenza A/California/04/2009 was optimized.HA,NP and 3 tandem M2e(3M2e)were cloned into pcDNA3.1 vector,respectively.Then the mRNAs were synthesized by linearization,in vitro transcription,enzymatic capping and enzymatic tailing,and named as mRNA-HA,mRNA-NP and mRNA-3M2e,respectively.The protein expression of the 3 kinds of mRNAs in 293T cells was detected by immunofluorescence assay.Comb-mRNA vaccine was prepared by enveloped mRNA-HA,mRNA-NP and mRNA-3M2e with lipid nanoparticles,respectively,and the particle size and potential were identified.Twenty-eight 6-week-old female BALB/c mice(18～22 g)were randomly divided into LNP group(n=14)and Comb-mRNA group(n=14).Hemagglutination inhibition(HI)method and microneutralization(MN)test were used to evaluate the serum antibody titer induced by Comb-mRNA vaccines.The mice were infected by 5LD50 wild-type H1 N1 influenza virus to evaluate the protective efficacy.Results The mRNA-HA,mRNA-NP and mRNA-3M2e were successfully constructed,and the 3 mRNAs could be expressed in 293T cells.The average size of mRNA encapsulated by lipid nanoparticles was 119.53±6.5 nm,and the average potential was-8.23±1.3 mV.The geometric mean titer(GMT)of HI and MN in the Comb-mRNA group were 179.6 and 201.6,compared with the LNP group.The ratio of IFN-γ+CD4+/CD8+Tcells was increased.The Comb-mRNA group could provide protection against 5LD50 wild type influenza H1 N1 virus after 2 weeks of booster immunization.Conclusion Comb-mRNA,an influenza vaccine candidate,can induce immune responses and protect mice from influenza virus challenge.

Objective To explore the effect of pristimerin combined with cisplatin on proliferation and apoptosis of nasopharyngeal carcinoma cells.Methods CCK-8 assay was used to examine the survival rate of HNE-1 and CNE-2Z cells following treatment for 24 h with different concentrations of pristimerin,cisplatin or their combination.The changes in colony formation ability,apoptosis,and intracellular reactive oxygen species(ROS)levels of the treated cells were analyzed using colony formation assay and flow cytometry.Western blotting was performed to detect the changes in protein expressions in the cells.The effects of pre-treatment with NAC on proliferation,apoptosis,and PI3K/AKT signaling pathway were observed in pristimerin-and/or cisplatin-treated cells.Results Both pristimerin and cisplatin significantly lowered the survival rate of HNE-1 and CNE-2Z cells(P<0.05).Compared with pristimerin or cisplatin alone,their combination more strongly inhibited survival and colony formation ability of the cells,increased cell apoptosis rate and intracellular ROS levels,upregulated the protein expressions of Bax,cleaved caspase-3,and cleaved PARP,and downregulated the protein expressions of Bcl-2,Mcl-1,PARP and p-PI3K and p-AKT(P<0.05).NAC pretreatment significantly attenuated proliferation inhibition and apoptosis-promoting effects of pristimerin combined with cisplatin,and partially restored the downregulated protein expressions of p-PI3K and p-AKT in HNE-1 and CNE-2Z cells with the combined treatment(P<0.05).Conclusion Pristimerin can enhance cisplatin-induced proliferation inhibition and apoptosis in nasopharyngeal carcinoma cells,the mechanism of which may involve ROS-mediated deactivation of the PI3K/AKT signaling pathway.

Objective To explore the correlation between serum microRNA-29a-3p(miR-29a-3p),throm-bospondin 2(THBS2)and cardiopulmonary function in children with chronic pulmonary heart disease.Meth-ods A total of 136 children with chronic pulmonary heart disease treated in Shijiazhuang Hospital of Tradi-tional Chinese Medicine from July 2019 to September 2023 were selected as the study subjects.Based on their clinical signs,cardiopulmonary function,and symptoms,they were divided into a compensated group(74 ca-ses)and a decompensated group(62 cases).Real-time fluorescence quantitative PCR(RT-qPCR)method was applied to detect serum miR-29a-3p level,and enzyme-linked immunosorbent assay(ELISA)was applied to detect serum THBS2 level.Moreover,ultrasound diagnostic equipment was applied to detect cardiac function indicators such as left ventricular ejection fraction(LVEF)and cardiac output(CO).Creatine kinase isoenzyme(CK-MB)and troponin(cTnl)were detected by electroluminescence analysis.Pulmonary function indicators were detected by pulmonary artery systolic pressure(PASP),mean pulmonary artery pressure(MPAP)and pulmonary artery diastolic pressure(PADP).Pearson correlation was used to analyze the corre-lation between serum miR-29a-3p and THBS2 levels and cardiopulmonary function indexes in children with chronic pulmonary heart disease,and multivariate Logistic regression analysis was used to screen the influen-cing factors of the disease in children with chronic pulmonary heart disease.Receiver operating characteristic(ROC)curve was drawn to analyze the evaluation value of serum miR-29a-3p and THBS2 levels in children with chronic pulmonary heart disease.Results The THBS2,CK-MB,cTnI,PASP,MPAP,and PADP in the decompensated group were higher than those in the compensated group(P＜0.05),while miR-29a-3p,LVEF,and CO were lower than those in the compensated group(P＜0.05).Serum miR-29a-3p in children with chro-nic pulmonary heart disease was positively correlated with LVEF and CO(P＜0.05),but negatively correla-ted with CK-MB,cTnI,PASP,MPAP,and PADP(P＜0.05).THBS2 was negatively correlated with LVEF and CO(P＜0.05),but positively correlated with CK-MB,cTnI,PASP,MPAP,and PADP(P＜0.05).MiR-29a-3p was a protective factor for exacerbation of chronic pulmonary heart disease in children(P＜0.05),while THBS2 was an independent risk factor for exacerbation of chronic pulmonary heart disease in children(P＜0.05).ROC curve analysis results showed that the area under the curve(AUC)of serum miR-29a-3p and THBS2 levels alone and in combination for evaluating the severity of chronic pulmonary heart disease in children were 0.827,0.817 and 0.915,respectively.Conclusion Serum miR-29a-3p and THBS2 levels are both factors affecting the disease of children with chronic pulmonary heart disease,and are closely related to the disease and cardiopulmonary function of children with chronic pulmonary heart disease.

Refractory prolactinoma is the most common pituitary neuroendocrine tumor.Dopamine receptor agonists(DA)are the primary choice for drug treatment.Most patients with prolactinomas respond well to DA.However,a minority of prolactinomas patients still show resistance to DA.Although drug-resistant and refractory prolactinomas are rare in clinical practice,their treatment is extremely challenging.Even a combination of drug therapy,multiple surgeries,and radiotherapy may not yield satisfactory outcomes.Therefore,standardizing the diagnosis and treatment process and pathway for refractory prolactionmas and exploring more effective multidisciplinary collaborative treatment strategies are urgent problems to be solved.In the clinical management of refractory prolactinomas,it is often necessary to consider the patient's condition comprehensively,replace other types of DA,or consider surgery,radiotherapy,and immunotherapy,which requires multidisciplinary diagnosis and treatment.This review synthesizes the latest literature at home and abroad to systematically discuss the latest advances in drug therapy,surgery,and radiotherapy treatments for refractory prolactionmas,aiming to provide new ideas for basic research,clinical diagnosis and treatment.

Objective To explore the effect of pristimerin combined with cisplatin on proliferation and apoptosis of nasopharyngeal carcinoma cells.Methods CCK-8 assay was used to examine the survival rate of HNE-1 and CNE-2Z cells following treatment for 24 h with different concentrations of pristimerin,cisplatin or their combination.The changes in colony formation ability,apoptosis,and intracellular reactive oxygen species(ROS)levels of the treated cells were analyzed using colony formation assay and flow cytometry.Western blotting was performed to detect the changes in protein expressions in the cells.The effects of pre-treatment with NAC on proliferation,apoptosis,and PI3K/AKT signaling pathway were observed in pristimerin-and/or cisplatin-treated cells.Results Both pristimerin and cisplatin significantly lowered the survival rate of HNE-1 and CNE-2Z cells(P<0.05).Compared with pristimerin or cisplatin alone,their combination more strongly inhibited survival and colony formation ability of the cells,increased cell apoptosis rate and intracellular ROS levels,upregulated the protein expressions of Bax,cleaved caspase-3,and cleaved PARP,and downregulated the protein expressions of Bcl-2,Mcl-1,PARP and p-PI3K and p-AKT(P<0.05).NAC pretreatment significantly attenuated proliferation inhibition and apoptosis-promoting effects of pristimerin combined with cisplatin,and partially restored the downregulated protein expressions of p-PI3K and p-AKT in HNE-1 and CNE-2Z cells with the combined treatment(P<0.05).Conclusion Pristimerin can enhance cisplatin-induced proliferation inhibition and apoptosis in nasopharyngeal carcinoma cells,the mechanism of which may involve ROS-mediated deactivation of the PI3K/AKT signaling pathway.

Objective:To prepare mesenchymal stem cells with antioxidant capacity (AO-MSC ) from human umbilical cords and evaluate its cell biological properties.Methods:In control group,mesenchymal stem cells (MSC) were isolated by digesting human umbilical cord Wharton's Jelly tissues with 0.2％ collagenase Ⅱ,and the released cells were collected and cultured in an animal serum-free culture medium.In AO-MSC group,incompletely collagenase Ⅱ-digested tissue debris were allowed to adhere to flusk flat bottoms and the AO-MSC was harvested by adherent culture. The conventional digestion and culture method was used as control.MSC colony forming ability was evaluated by fibroblast colony forming assay (CFU-F).MSC proliferative capacity was evaluated by CCK-8 assay.The MSC surface markers were detected by using flow cytometry and immunofluorescence staining.The adipogenic and osteogenic capacity of MSC was evaluated by multi-differentiation in vitro,and the mRNA expression of genes that control adipogenic and osteogenic differentiation was detected by real-time fluorescence quantitative PCR (RT-qPCR );Moreover,the mRNA expression of antioxidant substances such as SOD-1,GSH,GAT,and NQO1 in MSC was also evaluated by RT-qPCR.Results:The AO-MSC isolated by this strategy reached a confluence of 80％-90％ at around 18 days and grew in a swirling pattern.Flow cytometry and immunofluorescence staining assays showed that CD73,CD29,CD105,CD90 were highly expressed and CD31,CD45,HLA-DR were scarcely expressed in AO-MSC.AO-MSC exhibited stronger self-renewal and differentiation ability compared to MSC.However,the in vitro adipogenic-osteogenic capacity of MSC in the control group was stronger than that of AO-MSC.RT-qPCR assay showed that AO-MSC expressed higher mRNA levels of antioxidant substances compared to MSC.Conclusion:Human AO-MSC is successfully prepared from human umbilical cord without animal serum.

Objective:This study aims to establish an early warning diagnosis model for acute exacerbation of chronic obstructive pulmonary disease (AECOPD) and to provide a simple, rapid, and accurate auxiliary diagnosis basis for clinical practice.Methods:The sample bank of subjects (patients admitted to the Eighth Medical Center of the PLA General Hospital from September 10, 2021, to July 25, 2023) was constructed, including the model establishment cohort [SCOPD group 49, 42 males and 7 females, (69.71±11.16) years old; AECOPD group 53, 49 males and 4 females, (72.60±10.19) years old] and the model validation cohort [SCOPD group 35, 28 males and 7 females, (69.97±10.40) years old; AECOPD group 35, 33 males and 2 females, (71.43±9.67) years old]. Fasting peripheral blood samples were collected, and the expression levels of IL-5, IL-17A, and IFN-α were detected by flow cytometry. Different expression levels were analyzed by Mann-Whitney U test. Binary logistic regression analysis was used to screen the related risk factors of COPD patients in acute exacerbation. The diagnostic efficacy of the model was evaluated by the receiver operating characteristic (ROC) curve.Results:The levels of IL-5 [1.64 (0.60, 2.86) pg/ml], IL-17A [1.42 (0.88, 2.29) pg/ml], and IFN-α [0.91 (0.59, 1.81) pg/ml] in the SCOPD group were significantly decreased compared with the AECOPD group IL-5 [4.68 (2.34, 9.40) pg/ml, Z=-5.033, P<0.001], IL-17A [2.33 (1.59, 4.62) pg/ml, Z=-3.919, P<0.001], IFN-α [2.83 (0.91, 3.75) pg/ml, Z=-4.127, P<0.01] in the cohort of model establishment. The results of binary logistic regression analysis between SCOPD and AECOPD groups showed that IL-5, IL-17A, and IFN-α were independent risk factors for acute exacerbation of patients with COPD ( P<0.05). And the regression equation is Y=-2.861+0.364×IL-5+0.385×IL-17A+0.445×IFN-α. The AUC value of IL-5, IL-17A, IFN-α and combined detection was 0.866 ( P<0.001). Compared to the SCOPD group and the AECOPD group in the cohort of model validation, the receiver operating characteristic (ROC) curve showed that the combined model of three (AUC=0.858, P<0.001) could be used to diagnose the AECOPD. And the Kappa value was 0.773( P<0.05). Conclusion:The combined detection of IL-5, IL-17A, and IFN-α has high diagnostic efficacy for patients with acute exacerbation of COPD. This method provides a new potential tool for the clinical diagnosis of AECOPD and has the value of further exploration and optimization, promotion, and application.

Aim To explore the effect of Taohong Siwu Decoction on apoptosis and EMT of pulmonary fibrosis model rats by JAK2/STAT3 signaling pathway. Methods Forty SD rats were randomly divided into control group, model group, methylprednisolone group, Taohong Siwu Decoction low-concentration, high-concentration group respectively, with eight cases in each group. The intratracheal injection of bleomycin was applied to induce IPF rat models. HE and Masson staining were performed to observe the pathological changes of lung tissues in each group. ELISA was used to detect the contents of TNF-α, MMP-7 and TGFβ-l in serum of rats. Immunohistochemistry and Western blot were applied to detect the protein expression of JAK2, pJAK2, STAT3, p-STAT3, E-cadherin, α-SMA in lung tissues. RT-PCR was applied to detect the expression of JAK2, STAT3, Bcl2 and Bax genes in lung tissues. Results Compared with control group, the degree of alveolar inflammation and fibrosis degree, the contents of TNF-α, MMP-7 and TGFβ-1 in serum, the levels of JAK2, p-JAK2, STAT3, p-STAT3, α-SMA protein expression, JAK2, STAT3, Bax gene expression were up-regulated, and the levels of Bcl-2 gene and E-cadherin protein expression were down-regulated in lung tissues. Compared with model group, the degree of alveolar inflammation and fibrosis, the contents of TNF-α, MMP-7 and TGFβ-1 in serum, the levels of JAK2, p-JAK2, STAT3, p-STAT3, α-SMA protein expression, JAK2, STAT3 and Bax gene expression were reduced, while the levels of Bcl-2 gene and E-cadherin protein expression were elevated in Taohong Siwu Decoction high-concentration group. Conclusions Taohong Siwu Decoction may regulate JAK2/STAT3 signaling pathway, down-regulate Bax, α-SMA and up-regulate Bcl-2, E-cadherin expression to induce apoptosis and EMT in rat model of pulmonary fibrosis, thus playing an anti-pulmonary fibrosis role.

Aim To study the regulation and mechanism of phloroglucinol in bladder smooth muscle spasm. Methods In vitro the experiment used bladder muscle strip to verify the relieving effect of phloro-glucinol on bladder spasm by different drugs. At the same time,RT-qPCR and Western blot were used to detect the expression levels of genes involved in the calcium signaling pathway caused by the antispasmodic effect of phloroglucinol. Results Phloroglucinol could relieve bladder spasm, and the antispasmodic effect was enhanced with the increase of concentration, and the expression of calponin 1 and MYLK3 in tissue cells increased. The results of RT-qPCR showed that the expression of Gprc5b G,Ppp2r5a, Chptl, Prkar2b ,Abcd2 and Rasdl genes in mouse bladder tissue significantly decreased, which was consistent with the sequencing results of RNA-seq.Conclusions Phloroglucinol can relieve bladder smooth muscle spasm, and its mechanism is related to calcium signaling pathway. Meanwhile, phloroglucinol also inhibits the expression of Rasdl gene, suggesting that it may be related to cell cycle , protein phosphorylation, choline metabolism, ATP synthesis and tumor-related pathways.