Objective By simulating the etiology of abnormal uterine bleeding-ovulatory dysfunction(AUB-O)and establishing a rat model of abnormal uterine bleeding(AUB),this study aims to provide an experimental platform for investigating pathological mechanisms and developing therapeutic drugs for AUB.Methods After acclimation,24 adult(10-week-old)female SD rats were randomly divided into a normal control group(6 rats)and a model group(18 rats).The normal control group was housed in a barrier environment,while the model group underwent bilateral ovariectomy via dorsal approach in the same environment and rested for one week before starting to receive modeling drugs.In the model group,from Days 1 to 3 of modeling,each rat received a daily subcutaneous injection of 0.5 mg estradiol into the dorsal region.From Days 4 to 7,a daily subcutaneous injection of 5.0 mg progesterone was administered.On Day 6,rats received bilateral injections of 0.5 mL soybean oil per uterine cavity(total 1.0 mL)via the same dorsal surgical incision.On Day 8,mifepristone(10 mg/kg)was administered via oral gavage.The estrous cycle stage and its dynamic changes were continuously monitored during modeling.Uterine bleeding was recorded during the 48-hour observation period post-modeling.Serum and uterine tissue samples were collected from the model group at 0,12,24,36,and 48 h after mifepristone administration,while the normal control group was sampled at 36 h.The samples were subjected to HE staining,serum sex hormone ELISA,immunohistochemistry,TUNEL apoptosis staining,Western blotting,transcriptome sequencing,and bioinformatics analysis for comprehensive evaluation of the AUB rat model.Results The AUB rats exhibited uterine bleeding,endometrial detachment and injury,incomplete uterine restoration,inflammatory cell infiltration in the endometrium,enhanced tissue apoptosis,and structural damage of the stroma,glands,and vasculature.Compared with the normal control group,the levels of serum follicle-stimulating hormone(FSH),estradiol,and luteinizing hormone(LH)were significantly increased in the AUB rats(P＜0.05).The vascular density of the endometrium was significantly reduced(P＜0.05).The expression of vascular endothelial growth factor(VEGF)was qualitatively observed to be markedly enhanced at the site of endometrial detachment but significantly decreased around the stromal blood vessels(P＜0.01).Matrix metalloproteinase-9(MMP-9)expression was qualitatively observed to be strongly upregulated at the site of endometrial injury but significantly reduced in the non-detached stroma and glands(P＜0.01).Endometrial stromal cell apoptosis was significantly enhanced(P＜0.01).The expression levels of fibroblast growth factor 2(FGF2)and endothelin-1(ET-1)in uterine tissues were significantly decreased(P＜0.05).After comparing the transcriptome sequencing results of uterine tissues between AUB and normal rats,a total of 4 723 differentially expressed genes were identified,including 2 191 up-regulated genes and 2 532 down-regulated genes.KEGG enrichment analysis revealed that these differentially expressed genes were significantly enriched in pathways related to inflammation,immune apoptosis,cell signal transduction,proliferation and differentiation,and muscle contraction,among others.Conclusion An AUB rat model can be successfully established using a sequential administration protocol of estrogen,progesterone,and mifepristone to simulate the etiology of AUB-O.In this model,endometrial injury is associated with inflammation and apoptosis,with pathological manifestations influenced by abnormal vasoconstriction and impaired endometrial regeneration.This rat model closely recapitulates pathological characteristics of non-structural AUB observed in clinical practice,making it a validated experimental platform for exploring the pathological mechanisms and therapeutic interventions of non-structural AUB.

Objective:To explore the application effect of ADDIE model (analysis, design, development, implementation and evaluation) in the training of maxillofacial trauma first-aid ability of dental nurses, in order to optimize the training process of dental nurses.Methods:A self-controlled before and after study was conducted. Fifty-one dental nurses in Qingdao Stomatology Hospital Affiliated to Qingda University were selected as the research objects by convenient sampling method in March 2022, and the first aid ability training for maxillofacial trauma was carried out according to the five stages of ADDIE model. Before and after the training, the evaluation was made from three aspects: theory and skill test results, first-aid ability and training satisfaction scores.Results:Among 51 dental nurses, there were 4 males and 47 females, aged (30.69 ± 5.85) years. After the training, the theoretical assessment score of dental nursing staff was (88.87 ± 6.20) points, higher than that before the training (80.51 ± 7.21) points, and the technical assessment score was (91.61 ± 4.08) points, higher than that before the training (82.03 ± 7.56) points. The differences were statistically significant ( t = - 14.38, - 10.93, both P<0.01). The total score of first aid ability of nurses was (137.38 ± 11.30) points, higher than that before training (123.40 ± 13.73) points, and the difference was statistically significant ( t = - 17.30, P<0.01). The satisfaction score of dental nursing staff was (4.58 ± 0.50) points, higher than that before training (3.96 ± 0.46) points, and the difference was statistically significant ( t = - 11.51, P<0.01). Conclusions:The training program based on ADDIE model, through systematic teaching design, is helpful to improve the emergency treatment ability and training satisfaction of dental nursing staff with maxillofacial trauma.

Objective By simulating the etiology of abnormal uterine bleeding-ovulatory dysfunction(AUB-O)and establishing a rat model of abnormal uterine bleeding(AUB),this study aims to provide an experimental platform for investigating pathological mechanisms and developing therapeutic drugs for AUB.Methods After acclimation,24 adult(10-week-old)female SD rats were randomly divided into a normal control group(6 rats)and a model group(18 rats).The normal control group was housed in a barrier environment,while the model group underwent bilateral ovariectomy via dorsal approach in the same environment and rested for one week before starting to receive modeling drugs.In the model group,from Days 1 to 3 of modeling,each rat received a daily subcutaneous injection of 0.5 mg estradiol into the dorsal region.From Days 4 to 7,a daily subcutaneous injection of 5.0 mg progesterone was administered.On Day 6,rats received bilateral injections of 0.5 mL soybean oil per uterine cavity(total 1.0 mL)via the same dorsal surgical incision.On Day 8,mifepristone(10 mg/kg)was administered via oral gavage.The estrous cycle stage and its dynamic changes were continuously monitored during modeling.Uterine bleeding was recorded during the 48-hour observation period post-modeling.Serum and uterine tissue samples were collected from the model group at 0,12,24,36,and 48 h after mifepristone administration,while the normal control group was sampled at 36 h.The samples were subjected to HE staining,serum sex hormone ELISA,immunohistochemistry,TUNEL apoptosis staining,Western blotting,transcriptome sequencing,and bioinformatics analysis for comprehensive evaluation of the AUB rat model.Results The AUB rats exhibited uterine bleeding,endometrial detachment and injury,incomplete uterine restoration,inflammatory cell infiltration in the endometrium,enhanced tissue apoptosis,and structural damage of the stroma,glands,and vasculature.Compared with the normal control group,the levels of serum follicle-stimulating hormone(FSH),estradiol,and luteinizing hormone(LH)were significantly increased in the AUB rats(P＜0.05).The vascular density of the endometrium was significantly reduced(P＜0.05).The expression of vascular endothelial growth factor(VEGF)was qualitatively observed to be markedly enhanced at the site of endometrial detachment but significantly decreased around the stromal blood vessels(P＜0.01).Matrix metalloproteinase-9(MMP-9)expression was qualitatively observed to be strongly upregulated at the site of endometrial injury but significantly reduced in the non-detached stroma and glands(P＜0.01).Endometrial stromal cell apoptosis was significantly enhanced(P＜0.01).The expression levels of fibroblast growth factor 2(FGF2)and endothelin-1(ET-1)in uterine tissues were significantly decreased(P＜0.05).After comparing the transcriptome sequencing results of uterine tissues between AUB and normal rats,a total of 4 723 differentially expressed genes were identified,including 2 191 up-regulated genes and 2 532 down-regulated genes.KEGG enrichment analysis revealed that these differentially expressed genes were significantly enriched in pathways related to inflammation,immune apoptosis,cell signal transduction,proliferation and differentiation,and muscle contraction,among others.Conclusion An AUB rat model can be successfully established using a sequential administration protocol of estrogen,progesterone,and mifepristone to simulate the etiology of AUB-O.In this model,endometrial injury is associated with inflammation and apoptosis,with pathological manifestations influenced by abnormal vasoconstriction and impaired endometrial regeneration.This rat model closely recapitulates pathological characteristics of non-structural AUB observed in clinical practice,making it a validated experimental platform for exploring the pathological mechanisms and therapeutic interventions of non-structural AUB.

OBJECTIVE: To assess the value of combined detection strategies using multiple technologies for the genetic testing and prenatal diagnosis for pedigrees affected with Duchenne muscular dystrophy (DMD) for optimizing genetic counseling and reproductive guidance. METHODS: This study has involved 142 subjects from 65 suspected DMD families who had visited the First Affiliated Hospital of Chongqing Medical University from January 2018 to December 2023. A combination of multiple ligation-dependent probe amplification (MLPA), quantitative fluorescence PCR, and next-generation sequencing (NGS) was used. After confirming the genetic diagnosis of the probands, prenatal diagnosis was provided for carrier mothers. This study was approved by the Medical Ethics Committee of the hospital (Ethics No.: 2021-264). RESULTS: Among the 142 subjects tested, 73 cases of large deletions/duplications and 15 cases of small variants of the DMD gene were detected. The hotspot regions for the variants were exons 45 to 55. A total of 41 variant types were identified, of which 3 were previously unreported. In 19 families with suspected patients, 7 exonic deletions, 2 exonic duplications, and 3 small variants were identified. Prenatal diagnosis was performed on 48 fetuses from 46 families, revealing 16 affected male fetuses (including 12 with deletion variants, 2 with duplication variants, and 2 with small variants). Seven carrier females were identified among the 16 female fetuses (including 6 with deletions and 1 with duplication). Among the couples with an affected fetus, 16 had opted to terminate the pregnancy, while the parents of 32 fetuses had chosen to continue with the pregnancy. In families undergoing prenatal diagnosis, 53 (79.1%) pregnant women and their family members were found to carry mutations of the DMD gene. CONCLUSION The combined detection strategy of MLPA, qPCR, and NGS can encompass large deletions/duplications and small variants of the DMD gene, providing timely and accurate prenatal diagnosis for families affected by DMD. In conjunction with genetic counseling, this can effectively reduce the risk of producing affected offspring, which is crucial for the prevention and control of this disease.
Humans ; Muscular Dystrophy, Duchenne/diagnosis* ; Prenatal Diagnosis/methods* ; Female ; Male ; Pregnancy ; Pedigree ; Genetic Testing/methods* ; Dystrophin/genetics* ; Adult ; Genetic Counseling ; High-Throughput Nucleotide Sequencing/methods* ; Exons

Lysosomes represent a promising target for cancer therapy and reducing drug resistance. However, the short treatment time and low efficiency of lysosomal targeting have limited the application in lysosome-targeting anticancer drugs. In this study, we proposed an adhesive-bandage approach and synthesized a new lysosomal targeting drug, namely long-term lysosome-targeting anticancer drug (LLAD). It contains a SLC38A9-targeting covalently bound moiety and an alkaline component both to prolong the inhibition of SLC38A9 in lysosomes and alkalinize lysosomes. Upon short term and low-dose treatment of HeLa cells, at passage 0, with LLAD, it rapidly alkalinized lysosomes and also can be detected in lysosomes even at passage 15. LLAD induced apoptosis in HeLa cells through long-term lysosomal damage, and showed better long-term anticancer effect than cisplatin in vivo. Overall, our study paves the way for developing long-term lysosomal targeting drugs to treat cancer and overcome the drug resistance of cancer cells, and also provides a candidate drug, LLAD, for treating cancer.

Cholangiocarcinoma (CCA) is a fatal malignancy with steadily increasing incidence and poor prognosis. Since most CCA cases are diagnosed at an advanced stage, systemic therapies, including chemotherapy, radiotherapy, targeted therapy, and immunotherapy, play a crucial role in the management of unresectable CCA. The recent advances in targeted therapies and immunotherapies brought more options in the clinical management of unresectable CCA. This review depicts the advances of targeted therapies and immunotherapies for unresectable CCA, summarizes crucial clinical trials, and describes the efficacy and safety of different drugs, which may help further develop precision and individualization in the clinical treatment of unresectable CCA.
Humans ; Cholangiocarcinoma/drug therapy* ; Immunotherapy/methods* ; Bile Duct Neoplasms/drug therapy* ; Molecular Targeted Therapy/methods*

Objective:To explore the application effect of ADDIE model (analysis, design, development, implementation and evaluation) in the training of maxillofacial trauma first-aid ability of dental nurses, in order to optimize the training process of dental nurses.Methods:A self-controlled before and after study was conducted. Fifty-one dental nurses in Qingdao Stomatology Hospital Affiliated to Qingda University were selected as the research objects by convenient sampling method in March 2022, and the first aid ability training for maxillofacial trauma was carried out according to the five stages of ADDIE model. Before and after the training, the evaluation was made from three aspects: theory and skill test results, first-aid ability and training satisfaction scores.Results:Among 51 dental nurses, there were 4 males and 47 females, aged (30.69 ± 5.85) years. After the training, the theoretical assessment score of dental nursing staff was (88.87 ± 6.20) points, higher than that before the training (80.51 ± 7.21) points, and the technical assessment score was (91.61 ± 4.08) points, higher than that before the training (82.03 ± 7.56) points. The differences were statistically significant ( t = - 14.38, - 10.93, both P<0.01). The total score of first aid ability of nurses was (137.38 ± 11.30) points, higher than that before training (123.40 ± 13.73) points, and the difference was statistically significant ( t = - 17.30, P<0.01). The satisfaction score of dental nursing staff was (4.58 ± 0.50) points, higher than that before training (3.96 ± 0.46) points, and the difference was statistically significant ( t = - 11.51, P<0.01). Conclusions:The training program based on ADDIE model, through systematic teaching design, is helpful to improve the emergency treatment ability and training satisfaction of dental nursing staff with maxillofacial trauma.

Objective:To explore the diagnosis and treatment of adolescence-onset methylenetetrahydrofolate reductase (MTHFR) deficiency.Methods:This was a retrospective case study. Nine patients with adolescence-onset MTHFR deficiency were diagnosed at Peking University First Hospital from January 2016 to December 2022, and followed up for more than 1 year. Their general information, clinical manifestations, laboratory tests, cranial images, MTHFR gene variants, diagnosis, treatment, and outcome were analyzed retrospectively.Results:The 9 patients came from 8 families. They had symptoms at age of 8.0 years to 17.0 years and diagnosed at 9.0 years to 17.5 years. Eight were male and 1 was female. Two patients were brothers, the elder brother developed abnormal gait at 17.0 years; and the younger brother was then diagnosed at 15.0 years of age and treated at the asymptomatic stage, who was 18.0 years old with normal condition during this study. The main manifestations of the 8 symptomatic patients included progressive dyskinesia and spastic paralysis of the lower limbs, with or without intellectual decline, cognitive impairment and behavioral abnormalities. Totally, 15 variants of MTHFR gene were identified in the 9 patients, including 8 novel variants. Five patients had brain image abnormalities. Increased plasma total homocysteine level (65-221 μmol/L) was found in all patients, and decreased to 20-70 μmol/L after treatment with betaine and calcium folinate. Besides, the 8 symptomatic patients had their behavior and cognitive problems significantly improved, with a legacy of lower limb motor disorders.Conclusions:Late-onset MTHFR deficiency can occur in adolescence. The diagnosis is usually delayed because of non-specific clinical symptoms. The test of blood total homocysteine could be used as a selective screening test. Eight novel varients of MTHFR gene were identified. Timely treatment can improve clinical condition significantly, and pre-symptomatic treatment may prevent brain damage.

Objective:To investigate the clinical features and outcomes of adolescence-onset methylmalonic acidemia (MMA) and explore preventive strategies.Methods:This was a retrospective case analysis of the phenotypes, genotypes and prognoses of adolescence-onset MMA patients. There were 55 patients diagnosed in Peking University First Hospital from January 2002 to June 2023, the data of symptoms, signs, laboratory results, gene variations, and outcomes was collected. The follow-ups were done through WeChat, telephone, or clinic visits every 3 to 6 months.Results:Among the 55 patients, 31 were males and 24 were females. The age of onset was 12 years old (range 10-18 years old). They visited clinics at Tanner stages 2 to 5 with typical secondary sexual characteristics. Nine cases (16%) were trigged by infection and 5 cases (9%) were triggered by insidious exercises. The period from onset to diagnosis was between 2 months and 6 years. Forty-five cases (82%) had neuropsychiatric symptoms as the main symptoms, followed by cardiovascular symptoms in 12 cases (22%), kidney damage in 7 cases (13%), and eye disease in 12 cases (22%). Fifty-four cases (98%) had the biochemical characteristics of methylmalonic acidemia combined with homocysteinemia, and 1 case (2%) had the isolated methylmalonic acidemia. Genetic diagnosis was obtained in 54 cases, with 20 variants identified in MMACHC gene and 2 in MMUT gene. In 53 children with MMACHC gene mutation,1 case had dual gene variants of PRDX1 and MMACHC, with 105 alleles. The top 5 frequent variants in MMACHC were c.482G>A in 39 alleles (37%), c.609G>A in 17 alleles (16%), c.658_660delAAG in 11 alleles (10%), c.80A>G in 10 alleles (10%), c.567dupT and c.394C>T both are 4 alleles (4%). All patients recovered using cobalamin, L-carnitine, betaine, and symptomatic therapy, and 54 patients (98%) returned to school or work.Conclusions:Patients with adolescence-onset MMA may triggered by fatigue or infection. The diagnosis is often delayed due to non-specific symptoms. Metabolic and genetic tests are crucial for a definite diagnosis. Treatment with cobalamin, L-carnitine, and betaine can effectively reverse the prognosis of MMA in adolescence-onset patients.

Objective:To evaluate the value of left echography (LVO) in the diagnosis of false ventricular aneurysm complicated with mural thrombus.Methods:The clinical data of 10 patients with suspected pseudoventricular aneurysm examined by thoracic echocardiography (TTE) in Wuhan Asian Heart Hospital from January 2018 to March 2024 were retrospectively analyzed. All patients underwent LVO examination to further diagnose pseudoventricular tumor and whether it was complicated with mural thrombosis. Computed tomography angiography (CTA) or cardiac magnetic resonance (CMR) examination was used as the gold standard to analyze the diagnostic value of LVO in the diagnosis of pseudoventricular tumor.Results:Among the 10 suspected pseudoventricular tumors examined by TTE, LVO detected 6 cases of left ventricular pseudoaneurysm and 1 case of right ventricular pseudoaneurysm; CTA confirmed that 6 cases of left ventricular pseudoaneurysm detected by LVO were correctly diagnosed, 1 case of right ventricular pseudoaneurysm was misdiagnosed, CMR diagnosed right ventricular diverticula, LVO diagnosis accuracy was 6/7, and 4 cases of thrombi were detected. The detection rate was 4/4. The maximum transverse diameter of the tumor body of the communicating mouth/false ventricular aneurysm was 0.46±0.04. 1 patient underwent coronary artery bypass grafting and resection of false ventricular aneurysm. 1 patient underwent coronary artery interventional stent surgery; 4 routine conservative drug treatment, follow-up observation; One case of right ventricular diverticulum did not require special treatment.Conclusions:LVO contrast agent can clearly show the tumor body and location, measure the tumor entrance and size, and show mural thrombus. It is the first choice for the identification of false ventricular tumor. The diverticula was similar to the image of false ventricular aneurysm, and the sensitivity and specificity of right ventricular wall motion were higher in CMR than in LVO.