Chemotherapy is currently the mainstay of systemic management for triple-negative breast cancer (TNBC), but chemoresistance significantly impacts patient outcomes. Our research indicates that Doxorubicin (Dox)-resistant TNBC cells exhibit increased glycolysis and ATP generation compared to their parental cells, with this metabolic shift contributing to chemoresistance. We discovered that ALKBH3, an m¹A demethylase enzyme, is crucial in regulating the enhanced glycolysis in Dox-resistant TNBC cells. Knocking down ALKBH3 reduced ATP generation, glucose consumption, and lactate production, implicating its involvement in mediating glycolysis. Further investigation revealed that aldolase A (ALDOA), a key enzyme in glycolysis, is a downstream target of ALKBH3. ALKBH3 regulates ALDOA mRNA stability through m¹A demethylation at the 3'-untranslated region (3'UTR). This methylation negatively affects ALDOA mRNA stability by recruiting the YTHDF2/PAN2-PAN3 complex, leading to mRNA degradation. The ALKBH3/ALDOA axis promotes Dox resistance both in vitro and in vivo. Clinical analysis demonstrated that ALKBH3 and ALDOA are upregulated in breast cancer tissues, and higher expression of these proteins is associated with reduced overall survival in TNBC patients. Our study highlights the role of the ALKBH3/ALDOA axis in contributing to Dox resistance in TNBC cells through regulation of ALDOA mRNA stability and glycolysis.

Purpose To elucidate the clinicopathological characteristics of primary malignant giant cell tumor of bone(PMGCTB)with mainly osteosarcoma-like morphology.Meth-ods Clinicopathologic features of 7 cases of PMGCTB were ret-rospectively analyzed.Results Among 7 patients with PMGCTB,there were 4 females and 3 males,aged between 9 and 66 years(mean age 39.5 years,median age 35 years).The distal femur emerged as the most frequent site to be involved(3/6).The main clinical manifestations included pain and swelling at the original site of the tumor.Radiological findings indicated osteolytic lesions,often combined with sclerotic areas;most ca-ses showed cortical bone destruction and soft tissue masses(5/7).Histologically,the majority of tumors exhibited typical mor-phological features of osteosarcoma with a few or without osteo-clast-like multinucleated giant cells.Positive immunoreaction with H3F3A G34W was confirmed in 6 cases and with H3F3A G34V in 1 case.SATB2 and p63 were positive in all cases,p53 was proved to be wild type,the Ki67 proliferation index ranged approximately from 10％to 50％.H3F3A p.G34W mutation was detected in 6 cases and only 1 case harboring H3F3A p.G34V mutation.Conclusion PMGCTB is exceedingly rare and difficult for accurate diagnosis,especially for those with atypical morphological features.A comprehensive analysis involving ra-diological,immunophenotypic,and molecular detection is neces-sary to rule out other high-grade sarcomas.

Objective:To investigate the clinicopathological characteristics of giant cell tumor of bone (GCTB) in children.Methods:A total of 35 cases of GCTB diagnosed at Shanghai Sixth People′s Hospital Affiliated to Shanghai Jiaotong University School from 2016 to 2023 were collected, and a retrospective analysis of clinicopathological features and imaging findings was conducted.Results:Pediatric GCTB accounted for approximately 4.6% of total GCTB cases during the study period. There were 11 males and 24 females. The onset age ranged from 9 to 18 years (mean age 15 years, median age 16 years), with 8 cases (8/35, 22.9%) experiencing postoperative recurrence. Twenty-eight cases (28/35, 80%) primarily affected long bones, while 7 cases involved small or irregular bones. Imaging revealed osteolytic changes as the predominant feature, with 3 cases exhibited open physis, one of which had the tumor primarily at the diaphysis without crossing the physis. Histologically, pediatric GCTB resembled adult cases, characterized by mononuclear cells and osteoclast-like giant cells. Seven cases with denosumab treatment demonstrated degrees of giant cell disappearance, increased fibrous tissue and reactive bone proliferation in the stroma. One case was diagnosed as pediatric multicentric GCTB, and three cases as pediatric primary malignant GCTB, with malignant transformation into osteosarcoma. In all 35 cases, mutations in the H3F3A gene were identified, comprising 32 cases with H3.3 p.G34W mutations, one case with H3.3 p.G34V mutation, and 2 cases with H3.3 p.G34L mutations. Notably, the former two categories were successfully validated at the protein level through immunohistochemical staining, utilizing highly specific antibodies tailored for these mutation types: H3.3 p.G34W antibody and H3.3 p.G34V antibody. However, immunohistochemical staining was not available for the last category.Conclusions:Pediatric GCTB predominantly affects females and occurs primarily in long bones, mainly around the knee joint, the majority of tumors predominantly arise in the epiphysis and extend into the metaphysis; however, in cases where the epiphyseal plates are still unclosed, the tumors may be restricted to the metaphysis. Detection of H3F3A gene mutation is crucial for the diagnosis and differential diagnosis of pediatric GCTB.

Organoids can well simulate the heterogeneity of tumors,including tumor microenvironment and immune response,which helps to more accurately predict patient responses to drug and treatment effects.Organoids can be used for drug screening before drugs enter body,thus reducing the time and cost of clinical trials.However,research on tissue-organoids still faces some challenges,such as technical limitation and ethical issue.This review mainly introduces the progress in the research of breast cancer organoids,including the definition of organoids,development history,advantages and application in breast cancer research.

Objective:To clarify the potential correlation between biological changes of meninges in periodontitis mice and cognitive impairment by analyzing the biological changes of meninges in periodontitis mice using single-cell RNA sequencing.Methods:Thirty C57BL/6 mice were divided into two groups by using random number table method (15 mice in each group). Mice in the control group were locally administered 2% carboxyl methyl cellulose (CMC) without Porphyromonas gingivalis (Pg) on both buccal sides. A mixture of Pg W83 and 2% CMC was applied on both buccal sides in the experimental group mice three times a week, lasting for 16 weeks in total. The absorption of alveolar bone, locomotor activity and cognitive function, the activation of microglia and astrocytes in the cortex were observed and assessed. The mRNA expression levels of Occludin in meninges and brain were detected in two groups. Single-cell RNA sequencing data of meninges were processed by uniform manifold approximation and projection (UMAP). Differential genes expressions of endothelial cells were processed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. In addition, real-time fluorescence quantitative PCR (RT-qPCR) was used to verify the expressions of transcription activating factor 3 (Atf3) and apolpoprotein L domain-containing 1 (Apold 1). Results:Methylene blue staining found the distances of buccal and palatal cement-enamel junction-alveolar bone crest in experimental mice [(185.60±17.60), (206.90±13.37) μm] increased significantly compared with the control group [(135.33±9.57), (163.05±14.98) μm] ( t=5.02, P=0.002; t=4.37, P=0.005). Open field experiment showed the total distance and average speed of mice in the experimental group [(971.88±164.57) cm, (3.25±0.55) cm/s] were not statistically significant compared with the control group [(914.24±278.81) cm, (3.05±0.93) cm/s] ( t=0.65, P=0.525; t=0.65, P=0.520). The recognition index of the experimental group [(48.02±16.92) %] was lower than the control group [(66.27±17.90) %] ( t=2.40, P=0.027) by novel object recognition tests. Compared with the control group [(63.56±11.88) %], the alternation of experimental group [(50.99±14.17) %] was significantly decreased in Y maze tests ( t=2.33, P=0.030). Immunohistochemistry results showed microglia and astrocytes were activated in the cortex of experimental mice. Compared with the control group (1.02±0.25, 1.04±0.31), the relative mRNA expressions of Occludin decreased significantly in the meninges and brain of periodontitis mice, respectively (0.61±0.10, 0.64±0.20) ( t=3.47, P=0.010; t=2.66, P=0.024). By single-cell RNA sequencing, meninges cells were divided into 11 types, such as endothelial cells, fibroblasts, immune cells and so on. Endothelial cells were the main cell types in meninges [the control group: 26.47% (1 589/6 004), the experimental group: 26.26% (807/3 073)]. Compared with the control group [5.56% (334/6 004)], the percentage of granulocytes increased in the periodontitis mice [11.65% (358/3 073)]. Using clustering analysis to further focus on endothelial cells, GO enrichment analysis revealed differential genes were mainly related to angiogenesis, cell adhesion, apoptosis and so on. KEGG enrichment analysis revealed that differential genes were related to signaling pathways of interleukin-17, relaxin and so on. The relative mRNA expressions of Atf3 and Apold1 in meninges of periodontitis mice (0.42±0.24, 0.54±0.27) were significantly lower than the control group (1.03±0.26, 1.02±0.23) ( t=3.88, P=0.005; t=3.02, P=0.017). Conclusions:The mice chronically infected with Pg W83 occurred memory impairment, neuroinflammation and changes of barrier function. In the meninges of periodontitis mice, there were infiltration of immune cells and down-regulation expressions of Atf3 and Apold1 by single-cell RNA sequencing. Meningeal immunity and changes of barrier function may play an important role in the cognitive impairment caused by periodontitis.

Objective To analyze the prognostic value of serum microRNA(miRNA)-21-5p and miRNA-5189-5p expression levels in elderly patients with prostate cancer after laparoscopic radical prostatectomy(LRP).Methods A total of 213 elderly prostate cancer patients treated with LRP from January 2014 to Sep-tember 2018 in the First Affiliated Hospital of Hainan Medical College were retrospectively selected as the study objects.According to the 5-year follow-up outcome,the patients were divided into good prognosis group(87 cases)and poor prognosis group(126 cases).The preoperative clinical baseline data and the expression levels of serum miRNA-21-5p and miRNA-5189-5p were collected.Logistic regression analysis was used to screen the factors affecting the prognosis of patients.The prognostic prediction model was constructed with independent risk factors,and the predictive value was analyzed by drawing receiver operating characteristic(ROC)curve.Results Multivariate Logistic regression analysis showed that serum prostate specific antigen(PSA)level,positive incisal margin,and expression levels of miRNA-21-5p and miRNA-5189-5p were inde-pendent risk factors for the prognosis of elderly prostate cancer patients receiving LRP(P＜0.05).The com-bined prediction model was established based on independent risk factors,and the area under the curve of each factor alone predicting prognosis of elderly prostate cancer patients receiving LRP was less than that of four combined prediction(P＜0.05).Conclusion The expression levels of serum miRNA-21-5p and miRNA-5189-5p are of high value in predicting the prognosis of elderly prostate cancer patients after LRP.

Guillain-Barré syndrome (GBS) defines a kind of Immune-mediated acute inflammatory peripheral neuropathy. Miller-Fisher Syndrome (MFS) is a special variant of GBS, with mostly one-way course and rare clinical recurrence. Only a few recurrent cases have been reported in China. Here we report a case of a young male patient with double vision and progressive aggravation of limb numbness, acute onset, with symptoms of upper respiratory tract infection before onset, accompanied by pupil abnormalities and autonomic nervous dysfunction, who was was admitted to our hospital for similar symptoms 3 years ago and was improved by immunotherapy. The patient had a triad of “ataxia, areflexia and ophthalmoplegia”. Cerebrospinal fluid showed protein-cell separation. Serum anti-Sulfatides antibody IgM, anti-GT1a antibody IgG, anti-GQ1b antibody IgG and anti-GM3 IgM were positive. Recurrent MFS was diagnosed and the symptoms improved after immunotherapy. This case suggests that MFS is clinically heterogeneous, a few patients can present with relapse and generally have a better prognosis with immunotherapy. Pre-existing infection and anti-GQ1b antibody production may be predisposing factors for MFS recurrence.

To investigate the effect of adipose-derived stem cells (ADSCs) on radiation-induced skin injury in SD rats.
 Methods: Radioactive particles 192Ir were used to irradiate the left medial thigh skin of SD rats, and the irradiation dose was at 90 Gy. Then, the rats were randomly allocated into a control group and a treatment group (each n=9). After the irradiation, the control group was injected with 60 μL PBS and the treatment group was injected with 60 μL ADSCs in irradiated skin. The progress of skin damage and healing was observed and photographed every day. Twenty-eighth days after the irradiation, the irradiated skin tissue was taken from the left thigh, and then fixed with formaldehyde fixative solution. At the same time, the skin tissue of the corresponding part of the normal group (n=9) that was not irradiated was also taken. After sampling, embedding and slicing, immunohistochemical staining was used to compare the levels of α-smooth muscle actin (α-SMA), and HE staining was used to compare pathological features of the skin.
 Results: Radioactive particle 192Ir caused the development of III or IV radioactive skin damage. The score of the treatment group was significantly lower than that of the control group. The wounds of the treatment group were basically healed at 28 days, while the ulcer of the control group was unhealed. So, the healing time was shorter in the treatment group. The expression of α-SMA in the skin of the two groups was increased after the radiotherapy. By analyzing the pathological microstructure image, we found that the thickness of epidermis in the control group was greater than that in the treatment group, while the vascular density in the treatment group was greater than that in the control group (all P<0.05).
 Conclusion: Radioactive particles 192Ir can cause skin damage, while the adipose-derived stem cells might alleviate radiation-induced skin injury and promote ulcer healing by promoting angiogenesis.
Adipocytes ; Animals ; Iridium Radioisotopes ; Rats ; Rats, Sprague-Dawley ; Skin ; Stem Cells

Objective The study was designed to comparatively analyze ultrasonic appearance,laboratory and pathological findings in autoimmune hepatitis(AIH),and their correlation.The clinical value of ultrasound in diagnosing autoimmune hepatitis was studied.Methods By retrospectively reviewing the ultrasonic images,pathological and laboratory results of 68 patients with autoimmune hepatitis admitted to the PLA 302 Hospital from 2014 to 2015,we tried to reveal the correlation between ultrasonic features and pathological and laboratory findings.Results The ultrasonic diagnosis of liver fibrosis was not statistically correlated with the features of pathological ″interface hepatitis″ and serological liver function test.But it was positively correlated with the features of pathological ″spotty necrosis″(r=0.5099,P ＜ 0.001).The ultrasonic features of ″cord-like structure″ and ″nodular change″ had statistically improved the classification of the degree of AIH fibrosis on ultrasonic diagnosis(t=3.9547,P ＜ 0.01).The ultrasonic feature of the change of ″liver size″ and ″morphology″ also have statistically improved the diagnosis of AIH fibrosis with ultrasonography(t=2.070,2.137,4.584,3.773,all P＜0.05).Conclusion Ultrasonic images could provide objective evidence in diagnosis of AIH and evaluation of fibrosis degree.

Objective To explore the diagnostic value of Alzheimer-associated neuronal thread protein(AD7C-NTP)and olfactory function in the differentiation of three types of dementia,and to evaluate their clinical application value.Methods Mini-Mental State Examination (MMSE)and Montreal Cognitive Assessment(MoCA)were applied to evaluate cognitive function of all subjects with Alzheimer disease(AD),frontotemporal dementia (FTLD),or mixed dementia (MD).Enzyme-linked immunosorbent assay(ELISA)was used to detect the expression levels of AD7c-NTP in urine.T&T test method was applied to detect the olfactory function.Spearman rank correlation was used to evaluate the correlation of urine AD7c NTP with MMSE and MoCA scores.Results There was no significant difference in the demographic profile (except age)among three types of dementia of AD,FTLD and MD(F =4.05,P =0.02).Among the three dementia groups,the mean age of the MD group was highest.The statistically significant difference in MMSE scores was found among the three groups(F 3.79,P=0.03),while there was no significant difference inMoCAand NPI scores among the three dementia groups.The levels of the urine AD7c-NTP were different among the three dementia groups,but without statistical significance(H 1.25,P =0.53).Additionally,the FTLD group had the highest urine AD7c NTP level.Spearman rank correlation analysis showed no correlation of AD7c-NTP with MMSE and MoCA(r =0.18,P =0.25;r =0.14,P =0.39,respectively).No differences in olfactory function of the recognition domain(H =3.40,P=0.18)and in the detection domain(H =2.07,P=0.36)were found among three dementia groups of AD,FTLD and MD.Conclusions The level of urine AD7c-NTP is not of clinical significance in differentiating three types of dementia,and it is not correlated with the MMSE and MoCA scores.This study fails to find the clinical value of olfactory function test for distinguishing three types of dementia.