BACKGROUND: Temozolomide (TMZ) resistance is a significant challenge in treating glioblastoma (GBM). Collagen remodeling has been shown to be a critical factor for therapy resistance in other cancers. This study aimed to investigate the mechanism of TMZ chemoresistance by GBM cells reprogramming collagens. METHODS: Key extracellular matrix components, including collagens, were examined in paired primary and recurrent GBM samples as well as in TMZ-treated spontaneous and grafted GBM murine models. Human GBM cell lines (U251, TS667) and mouse primary GBM cells were used for in vitro studies. RNA-sequencing analysis, chromatin immunoprecipitation, immunoprecipitation-mass spectrometry, and co-immunoprecipitation assays were conducted to explore the mechanisms involved in collagen accumulation. A series of in vitro and in vivo experiments were designed to assess the role of the collagen regulators prolyl 4-hydroxylase subunit alpha 1 (P4HA1) and yes-associated protein (YAP) in sensitizing GBM cells to TMZ. RESULTS: This study revealed that TMZ exposure significantly elevated collagen type I (COL I) expression in both GBM patients and murine models. Collagen accumulation sustained GBM cell survival under TMZ-induced stress, contributing to enhanced TMZ resistance. Mechanistically, P4HA1 directly binded to and hydroxylated YAP, preventing ubiquitination-mediated YAP degradation. Stabilized YAP robustly drove collagen type I alpha 1 ( COL1A1) transcription, leading to increased collagen deposition. Disruption of the P4HA1-YAP axis effectively reduced COL I deposition, sensitized GBM cells to TMZ, and significantly improved mouse survival. CONCLUSION P4HA1 maintained YAP-mediated COL1A1 transcription, leading to collagen accumulation and promoting chemoresistance in GBM.
Temozolomide ; Humans ; Glioblastoma/drug therapy* ; Animals ; Mice ; Cell Line, Tumor ; Drug Resistance, Neoplasm/genetics* ; YAP-Signaling Proteins ; Hydroxylation ; Dacarbazine/pharmacology* ; Adaptor Proteins, Signal Transducing/metabolism* ; Transcription Factors/metabolism* ; Collagen/biosynthesis* ; Collagen Type I/metabolism* ; Prolyl Hydroxylases/metabolism* ; Antineoplastic Agents, Alkylating/therapeutic use*

Cemental tear is a rare and indetectable condition unless obvious clinical signs present with the involvement of surrounding periodontal and periapical tissues. Due to its clinical manifestations similar to common dental issues, such as vertical root fracture, primary endodontic diseases, and periodontal diseases, as well as the low awareness of cemental tear for clinicians, misdiagnosis often occurs. The critical principle for cemental tear treatment is to remove torn fragments, and overlooking fragments leads to futile therapy, which could deteriorate the conditions of the affected teeth. Therefore, accurate diagnosis and subsequent appropriate interventions are vital for managing cemental tear. Novel diagnostic tools, including cone-beam computed tomography (CBCT), microscopes, and enamel matrix derivatives, have improved early detection and management, enhancing tooth retention. The implementation of standardized diagnostic criteria and treatment protocols, combined with improved clinical awareness among dental professionals, serves to mitigate risks of diagnostic errors and suboptimal therapeutic interventions. This expert consensus reviewed the epidemiology, pathogenesis, potential predisposing factors, clinical manifestations, diagnosis, differential diagnosis, treatment, and prognosis of cemental tear, aiming to provide a clinical guideline and facilitate clinicians to have a better understanding of cemental tear.
Humans ; Dental Cementum/injuries* ; Consensus ; Diagnosis, Differential ; Cone-Beam Computed Tomography ; Tooth Fractures/therapy*

Objective The purpose of this study was to investigate the effects of OTU deubiquitinase,ubiquitin al-dehyde binding 2(OTUB2)on the activity of DEAD-box helicase 54(DDX54)and its influence on the formation of neutrophil extracellular traps(NETs)and the vitality and invasion of colorectal cancer(CRC)cells.Methods Peripheral blood neutrophils were isolated from CRC patients and healthy controls,then stimulated with phorbol-12-myristate-13-acetate(PMA)or DNaseⅠfor 4 hours.Western blot analysis was performed to detect the expression of NETs markers,myeloperoxidase(MPO),and citrullinated histone H3(Cit-H3).The expression of OTUB2 or DDX54 was manipulated in SW480 cells,and they were co-cultured with neutrophils.The cells were divided into the following groups:Control group(without any treatment),NETs group,vector group,OTUB2 group,NETs+si-OTUB2 group(SW480 cells,SW480 cells transfected with vector,OTUB2 overexpression plasmid and si-OTUB2 were co-cultured with PMA-treated neutrophils,respectively),OTUB2+DNase Ⅰ group(SW480 cells transfected with OTUB2 overexpression plasmid co-cultured with neutrophils treated with DNase Ⅰ),si-OTUB2group,OTUB2+si-NCgroup,OTUB2+si-DDX54group(SW480 cells transfected with si-OTUB2,OTUB2 overexpressing plasmid and si-NC,OTUB2 overexpressing plasmid and si-DDX54 were co-cultured with neutro-phils,respectively).ELISA was used to measure the relative expression of MPO-DNA complexes and the concen-tration of Cit-H3 in the supernatant of SW480 cells.MTT and Transwell assays were performed to evaluate cell via-bility and invasive ability.RNA sequencing was conducted to screen downstream genes regulated by OTUB2.The interaction between DDX54 and OTUB2 was validated using quantitative real-time polymerase chain reaction(qRT-PCR),co-immunoprecipitation(Co-IP),Glutathione S-Transferase(GST)pull-down,and His-tag pull-down ex-periments.Results Compared to healthy individuals,there was an increased formation of NETs in the peripheral blood of CRC patients.The CRC cells in the NETs group exhibited increased cell viability and invasion compared to the control group(P<0.05).In comparison to the vector group,the OTUB2 group showed increased relative ex-pression of MPO-DNA complexes and Cit-H3,as well as increased cell viability and invasion(P<0.05).Howev-er,when compared to the OTUB2 group,the OTUB2+DNase Ⅰ group exhibited decreased relative expression of MPO-DNA complexes and Cit-H3,as well as decreased cell viability and invasion(P<0.05).Co-IP,GST pull-down,and His-tag pull-down experiments demonstrated the interaction between OTUB2 and DDX54.In comparison to the OTUB2+si-NC group,the OTUB2+si-DDX54 group showed decreased relative expression of MPO-DNA complexes and Cit-H3,as well as decreased cell viability and invasion(P<0.05).Conclusion deubiquitinating enzyme OTUB2 can increase the formation of NETs and promote CRC cell viability and invasion by up-regulating DDX54.

Objective A comparison of different routes for the administration of clodronate disodium liposomes to determine the most effective method of rat endometrial macrophage clearance.Methods Female 8-week-old SD rats were randomly divided into a unilateral control group(injected with 100 μL PBS liposomes into the left uterine cavity),unilateral clearance group(injected with 100 μL clodronate liposomes into the right uterine cavity),bilateral control group(injected with 100 μL PBS liposomes into the bilateral intrauterine),bilateral clearance group(injected with 100 μL clodronate liposomes into the bilateral intrauterine),whole-body control group(injected with 500 μL PBS liposomes into the caudal vein),and whole-body clearance group(injected with 500 μL clodronate liposomes into the caudal vein).Hematoxylin and eosin staining was used to observe the morphology and structures of uterine and ovarian tissues,immunohistochemistry was used to observe the presence of macrophages in uterine and ovarian tissues,and flow cytometry was used to detect changes to macrophage cell counts in uterine and ovarian tissues.Results There were no significant differences in the structures or morphology of the uterus and ovary among the groups.Immunohistochemical staining showed that,compared to the control group,the unilateral and bilateral uterine clearance groups'population of terine macrophages was significantly decreased(P＜0.001),but there was no difference in the accumulation of macrophages in the ovary(P＞0.05).The number of macrophages in both uterine and ovarian tissues decreased in the whole-body clearance group(P＜0.01,P＜0.01).Compared with the unilateral and bilateral clearance groups,the whole-body clearance group had more macrophages in the ovarian tissues(P＜0.05).Flow cytometry showed that,compared with the control group,each clearance group's percentages of macrophages in the uterine tissue were significantly reduced(P＜0.001,P＜0.001,P＜0.05).Compared to the whole-body clearance group,the unilateral and bilateral clearance groups'endometrial macrophages had superior clearance activity(P＜0.05,P＜0.05).In addition,the number of macrophages in ovarian tissue decreased in all clearance groups compared to the control group.The decrease in macrophage number was most pronounced in the whole-body clearance group(P＜0.05),and there was no significant difference in numbers between the unilateral and bilateral clearance groups and the control group(P＞0.05).Conclusions Local injection of clodronate liposomes was more effective than caudal injection for clearing uterine macrophages,and the impact on ovarian macrophages was largely avoided.Thus local clodronate liposome injection is an improved method for establishing a local uterine macrophage clearance model.

Objective To explore the regulatory effect of miR-6838-5p on DDR1 gene expression and proliferation of breast cancer cells.Methods The expression levels of miR-6838-5p in normal breast epithelial cells and breast cancer cells were detected using qRT-PCR,and the potential target genes of miR-6838-5p was predicted using TargetscanV 8.0.Double luciferase reporter gene experiment was performed to verify the binding between miR-6838-5p and DDR1.Breast cancer MCF-7 cells were transfected via liposome,miR-6838-5p mimic,miR-6838-5p inhibitor,DDR1 siRNA,DDR1-overexpresisng vector,or both miR-6838-5p mimic and DDR1-overexpressing vector,and the changes in cell proliferation were examined with CCK-8 and EdU assays;Western blotting was used to detect the expression of DDR1.The mediating role of DDR1 in miR-6838-5p overexpression-induced inhibition of MCF-7 cell proliferation was verified in a nude mouse model bearing MCF-7 cell xenografts.Results The expression of miR-6838-5p was significantly lower in breast cancer cells than in normal breast epithelial cells.In MCF-7 cells,miR-6838-5p overexpression induced significant inhibition of cell proliferation.Dual luciferase reporter gene experiment demonstrated a binding relationship between miR-6838-5p and DDR1(P<0.01).Western blotting showed that miR-6838-5p overexpression significantly lowered DDR1 expression in MCF-7 cells,and DDR1 overexpression promoted proliferation of the cells;co-transfection of the cells with DDR1-overexpressing vector significantly attenuated the inhibitory effect of miR-6838-5p mimic on cell proliferation.In the tumor-bearing nude mice,the xenografts overexpressing miR-6838-5p showed a significantly smaller volum with obviously the expression of DDR1.Conclusion Overexpression of miR-6838-5p inhibits breast cancer cell proliferation by regulating DDR1 expression.

AIM:To investigate the molecular mechanism of a novel bromobenzene substituted trifluoromethylbenzo Cyclopentanone WW02 inhib-iting the viability and proliferation of human lung cancer A549 and H1299 cells.METHODS:The ef-fect of different concentrations of WW02(6.25,12.5,25,50 μg/mL)on cell viability and prolifera-tion of A549 and H1299 were measured using CCK-8 and EdU methods.After 24 hours of stimulation of A549 and H1299 cells with different concentra-tions of WW02,the changes in Akt and mTOR phos-phorylation levels under different concentrations of WW02 were detected through Western blot as-say.Macromolecular docking was carried out be-tween WW02,AKT and mTOR through MOE Dock.RESULTS:After treating A549 and H1299 cells with WW02 using different concentrations(6.25,12.5,25,50 μg/mL),the activity of A549 and H1299 cells decreased in a concentration dependent manner compared with the DMSO control group(P<0.05).The proliferation of cells showed a concentration dependent decrease compared to the DMSO con-trol group(P<0.05).Compared with the DMSO con-trol group,after 24 hours of WW02 stimulation,the phosphorylation levels of Akt and mTOR in A549 cells decreased under the concentration of WW02(12.5,25,50 μg/mL,P<0.05).Compared with the DMSO control group,the phosphorylation levels of Akt and mTOR in H1299 cells decreased af-ter 24 hours of WW02 stimulation(25,50 μg/mL,P<0.05).Based on pattern analysis,it was found that WW02 had a strong binding with Akt and mTOR,with the highest score of-8.3 kcal/mol for WW02 and mTOR,while the highest score for WW02 and Akt was-7.3 kcal/mol.CONCLUSION:WW02 inhib-its the activity and proliferation of lung cancer A549 and H1299 cells,and its mechanism of action may be achieved by directly binding to Akt and mTOR proteins to inhibit Akt and mTOR phosphory-lation.

Objective To explore the regulatory effect of miR-6838-5p on DDR1 gene expression and proliferation of breast cancer cells.Methods The expression levels of miR-6838-5p in normal breast epithelial cells and breast cancer cells were detected using qRT-PCR,and the potential target genes of miR-6838-5p was predicted using TargetscanV 8.0.Double luciferase reporter gene experiment was performed to verify the binding between miR-6838-5p and DDR1.Breast cancer MCF-7 cells were transfected via liposome,miR-6838-5p mimic,miR-6838-5p inhibitor,DDR1 siRNA,DDR1-overexpresisng vector,or both miR-6838-5p mimic and DDR1-overexpressing vector,and the changes in cell proliferation were examined with CCK-8 and EdU assays;Western blotting was used to detect the expression of DDR1.The mediating role of DDR1 in miR-6838-5p overexpression-induced inhibition of MCF-7 cell proliferation was verified in a nude mouse model bearing MCF-7 cell xenografts.Results The expression of miR-6838-5p was significantly lower in breast cancer cells than in normal breast epithelial cells.In MCF-7 cells,miR-6838-5p overexpression induced significant inhibition of cell proliferation.Dual luciferase reporter gene experiment demonstrated a binding relationship between miR-6838-5p and DDR1(P<0.01).Western blotting showed that miR-6838-5p overexpression significantly lowered DDR1 expression in MCF-7 cells,and DDR1 overexpression promoted proliferation of the cells;co-transfection of the cells with DDR1-overexpressing vector significantly attenuated the inhibitory effect of miR-6838-5p mimic on cell proliferation.In the tumor-bearing nude mice,the xenografts overexpressing miR-6838-5p showed a significantly smaller volum with obviously the expression of DDR1.Conclusion Overexpression of miR-6838-5p inhibits breast cancer cell proliferation by regulating DDR1 expression.

Objective:To study the iodine nutritional status of pregnant women in the third trimester in the Southwest of Shandong Province, analyze its impact on their cardiac electrical activity, and provide a basis for scientific supplementation of iodine during pregnancy.Methods:From January 2021 to June 2022, a cross-sectional survey was conducted using cluster random sampling method. According to the inclusion and exclusion criteria, 200 pregnant women in the third trimester were selected from 3 tertiary hospitals in three cities in the Southwest of Shandong Province, and were divided into the third trimester group ( n = 600), and 100 non-pregnant women were selected as the control group ( n = 300). The urinary iodine content was detected by arsenic-cerium catalytic spectrophotometry, and the pregnant women in the third trimester group were subdivided into iodine deficiency subgroup [G1 subgroup, median urinary iodine (MUIC) < 150 μg/L] based on the MUIC, iodine excess subgroup (G2 subgroup, MUIC≥500 μg/L) and moderate iodine subgroup (G3 subgroup, 150 μg/L≤MUIC < 500 μg/L). Chemiluminescence immunoassay was used to measure the serum levels of thyroid-stimulating hormone (TSH), free thyroxine (FT 4) and free triiodothyronine (FT 3). The cardiac electrical indexes were detected by a 12-lead surface electrocardiogram (ECG) machine. Results:There was no statistically significant difference in urinary iodine levels between pregnant women in the third trimester group and non-pregnant women in the control group among the 3 tertiary hospitals in the Southwest of Shandong Province ( H = 3.63, 3.27, P > 0.05). In the third trimester group, the proportion of pregnant women in the G1, G2 and G3 subgroups was 27.67% (166/600), 6.83% (41/600) and 65.50% (393/600), respectively. There was a statistically significant difference in urinary iodine levels between the subgroups and the control group ( H = 11.56, P < 0.001). The serum FT 3 and FT 4 levels in the G2 subgroup were lower than those in the G1 and G3 subgroups ( P < 0.001), but there was no statistically significant difference in serum TSH levels among the three subgroups ( P > 0.05). The normal rates of ECG in the G1, G2, G3 subgroups, and the control group were 38.55% (64/166), 41.46% (17/41), 92.37% (363/393), and 95.33% (286/300), respectively. The difference between the groups were statistically significant (χ 2 = 461.25, P < 0.001), the normal rate of ECG in the G1 and G2 subgroups was lower than that in the control group ( P < 0.001). Short P-R intervals and ST-T changes were the most common abnormal ECG in the third trimester group. Conclusions:The incidence of iodine deficiency, iodine excess, and other abnormal iodine nutritional status in pregnant women in the third trimester of the Southwest of Shandong Province is relatively high. Short P-R intervals, ST-T changes, and other arrhythmia caused by this are more common. It is necessary to strengthen monitoring of iodine nutritional status and ECG during pregnancy, and adjust intervention strategies such as iodine supplementation in a timely manner.

As the second largest vector of infectious diseases, ticks carry and transmit various pathogens that cause human infections and pose a serious public health hazard. In recent years, there has been an ongoing epidemic of tick-borne fever with thrombocytopenia syndrome virus (SFTSV), as well as the occurrence of human infections by brand-new viruses such as Jingmen tick virus (JMTV) and Alongshan virus (ALSV) in China. This paper will review the advancement of disease clinical characteristics, laboratory methods of virus isolation, immunology and molecular biology of these emerging tick-borne viral diseases in China.

Objective:To explore the clinical value of synchronized 12-lead Holter in the diagnosis of coronary heart disease myocardial ischemia.Methods:A total of 101 patients with coronary heart disease who came to the Affiliated Hospital of Jining Medical College from May 2019 to May 2020 were selected. They all received conventional electrocardiogram(ECG) and synchronized 12-lead Holter examinations. The value of different examination methods in the diagnosis of myocardial ischemia was compared, and the characteristics of myocardial ischemia in patients with different degrees of coronary artery disease were analyzed.Results:The detection rates of synchronized 12-lead Holter for myocardial ischemia in single-vessel coronary artery disease and multi-vessel coronary artery disease were 28.71% (29/101) and 56.44%(57/101), respectively, which were slightly higher than 17.82%(18/101) and 45.54% (46/101) of conventional ECG, but with no statistically significant difference ( P>0.05), the total myocardial ischemia detection rate of synchronized 12-lead Holter was 85.15%(86/101), which was higher than 63.37%(64/101) of conventional ECG ( P<0.05). The mean frequency of myocardial ischemia in coronary heart disease diagnosed by synchronized 12-lead Holter was higher than that of conventional ECG, and the duration and amplitude of ST-segment depression were higher than those in conventional ECG ( P<0.05). The detection rates of lateral and inferior myocardial ischemia on synchronized 12-lead Holter were 16.28%(14/86) and 22.09%(19/86), respectively, which were higher than 0 of conventional ECG. The detection rate of myocardial ischemia in the anterior septum/anterior wall was 61.62%(53/86), which was lower than 100.00% (64/64)of the conventional ECG ( P<0.01). Synchronized 12-lead Holter in the diagnosis of coronary cardiac myocardial ischemia was highly consistent with that by coronary angiography, the Kappa value was 0.648, and the sensitivity, specificity, accuracy and negative predictive value were 93.33%, 81.82%, 92.08% and 60.00%, respectively, which were higher than 54.44%, 63.64%, 55.45%, 14.58% of conventional ECG ( P<0.05). The number of myocardial ischemic attacks in patients with multi-vessel disease of coronary heart disease was more than that of single vessel disease ( P<0.05), the mean frequency of myocardial ischemia was higher than that of single vessel disease, and the duration and amplitude of ST-terminal depression were higher than those of patients with single-vessel disease ( P<0.05). Conclusions:Synchronized 12-lead Holter is more effective than conventional ECG in the diagnosis of myocardial ischemic attack of coronary heart disease, and it is more consistent with coronary angiography. It can clarify the frequency and extent of myocardial ischemic attacks and help identify the location of myocardial ischemia. It can be used as an important basis for screening myocardial ischemia attacks of coronary heart disease.