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.

Objective:To explore the technical points and precautions of using bilateral free posteromedial thigh flap to reconstruct bilateral breasts.Methods:A retrospective analysis was conducted on the clinical data of patients who underwent bilateral mastectomy and bilateral free posteromedial thigh flap transplantation for bilateral breast reconstruction at Hunan Cancer Hospital from June 2020 to January 2023. The operation was carried out simultaneously by two groups of doctors. Breast surgeons performed modified radical mastectomy for breast cancer or subcutaneous mastectomy. The flap group prepared bilateral free posteromedial thigh flaps. The posteromedial thigh flaps were designed in three patterns: transverse, vertical and oblique. The intrathoracal vessels were used as recipient vessels in all cases. There were two types of vascular anastomosis method: ① For flaps with single vascular pedicle, the artery was anastomosed with the proximal end of the internal thoracic artery, and the only accompanying vein was anastomosed with the proximal end of the internal thoracic vein, or the two accompanying veins were anastomosed with the proximal and distal ends of the internal thoracic vein; ② For flaps with dual vascular pedicles, arteries were anastomosed with the proximal and distal ends of the internal thoracic artery, and the accompanying veins on both sides communicated with the proximal and distal ends of the internal thoracic vein, respectively. Normal distribution measurement data were expressed as Mean±SD.Results:A total of 9 female patients were enrolled, aged 29 to 43 years, with an average age of 38.3 years. Among them, bilateral breast cancer was considered in 4 cases, unilateral breast cancer with multiple nodules in the opposite breast was in 3 cases, and bilateral multiple nodules along with high-risk of developing breast cancer suggested by gene testing were found in 2 cases. Modified radical mastectomy for breast cancer and nipple-sparing subcutaneous mastectomy were performed in 7 and 11 breasts, respectively. There were three design method for the posteromedial thigh flap: horizontal design (6 thighs); vertical design (6 thighs); diagonal design (6 thighs). A total of 18 posteromedial thigh flaps were raised. Four different types of posteromedial thigh flaps were used: gracilis muscle flap (6 thighs); great adductor muscle perforator flap (5 thighs); great adductor muscle flap (5 thighs); femoral gracilis flap+ adductor magnus flap (2 thighs). The skin flaps harvested were (18.9±0.6) cm in length, (7.2±0.4) cm in width and (4.2±0.3) cm in thickness. The average weight of the skin flaps were 235 grams (185-355 grams). The aforementioned vascular anastomosis method ① and ② were carried out in 15 and 3 breasts, respectively. The patients were followed up for 6 to 32 months, with an average of 17.3 months. All skin flaps survived, and the reconstructed breasts presented with good appearance, good elasticity, and no skin flap contraction or deformation occurred; scarring on the donor area was linear and mild; there was no significant impact on lower limb movement. The breast shape was satisfactory, and there was no recurrence of breast cancer.Conclusion:The application of bilateral free posteromedial thigh flap transplantation for reconstruction of bilateral breasts has flexible flap preparation and vascular anastomosis forms, which can achieve satisfactory result of bilateral symmetry.

Objective:To explore the technical points and precautions of using bilateral free posteromedial thigh flap to reconstruct bilateral breasts.Methods:A retrospective analysis was conducted on the clinical data of patients who underwent bilateral mastectomy and bilateral free posteromedial thigh flap transplantation for bilateral breast reconstruction at Hunan Cancer Hospital from June 2020 to January 2023. The operation was carried out simultaneously by two groups of doctors. Breast surgeons performed modified radical mastectomy for breast cancer or subcutaneous mastectomy. The flap group prepared bilateral free posteromedial thigh flaps. The posteromedial thigh flaps were designed in three patterns: transverse, vertical and oblique. The intrathoracal vessels were used as recipient vessels in all cases. There were two types of vascular anastomosis method: ① For flaps with single vascular pedicle, the artery was anastomosed with the proximal end of the internal thoracic artery, and the only accompanying vein was anastomosed with the proximal end of the internal thoracic vein, or the two accompanying veins were anastomosed with the proximal and distal ends of the internal thoracic vein; ② For flaps with dual vascular pedicles, arteries were anastomosed with the proximal and distal ends of the internal thoracic artery, and the accompanying veins on both sides communicated with the proximal and distal ends of the internal thoracic vein, respectively. Normal distribution measurement data were expressed as Mean±SD.Results:A total of 9 female patients were enrolled, aged 29 to 43 years, with an average age of 38.3 years. Among them, bilateral breast cancer was considered in 4 cases, unilateral breast cancer with multiple nodules in the opposite breast was in 3 cases, and bilateral multiple nodules along with high-risk of developing breast cancer suggested by gene testing were found in 2 cases. Modified radical mastectomy for breast cancer and nipple-sparing subcutaneous mastectomy were performed in 7 and 11 breasts, respectively. There were three design method for the posteromedial thigh flap: horizontal design (6 thighs); vertical design (6 thighs); diagonal design (6 thighs). A total of 18 posteromedial thigh flaps were raised. Four different types of posteromedial thigh flaps were used: gracilis muscle flap (6 thighs); great adductor muscle perforator flap (5 thighs); great adductor muscle flap (5 thighs); femoral gracilis flap+ adductor magnus flap (2 thighs). The skin flaps harvested were (18.9±0.6) cm in length, (7.2±0.4) cm in width and (4.2±0.3) cm in thickness. The average weight of the skin flaps were 235 grams (185-355 grams). The aforementioned vascular anastomosis method ① and ② were carried out in 15 and 3 breasts, respectively. The patients were followed up for 6 to 32 months, with an average of 17.3 months. All skin flaps survived, and the reconstructed breasts presented with good appearance, good elasticity, and no skin flap contraction or deformation occurred; scarring on the donor area was linear and mild; there was no significant impact on lower limb movement. The breast shape was satisfactory, and there was no recurrence of breast cancer.Conclusion:The application of bilateral free posteromedial thigh flap transplantation for reconstruction of bilateral breasts has flexible flap preparation and vascular anastomosis forms, which can achieve satisfactory result of bilateral symmetry.

Objective To explore the effect of young plasma intraperitoneal injection on the fertility and ovarian function of aging mice and analyze its potential molecular mechanism.Methods Fifty-four-week-old female C57BL/6 mice and 8-week-old male C57BL/6 mice were selected.Among them,the female mice were randomly divided into three groups:the young plasma group,the aging plasma group and the normal saline group.The young plasma group and the aging plasma group received intraperitoneal injection of plasma from young(25-29 years old)and elderly(45-49 years old)female donors,respectively.Each injection was 500 μL,administered every other day for 2 weeks.The saline group received an equal volume of saline.After the last injection,mating experiments were conducted to evaluate fertility.Ovarian histopathological changes were observed by HE staining.Oocytes and fertilized eggs were collected after superovulation and cultured in vitro to assess oocyte quality and embryo developmental potential.Transcriptomic analysis of ovarian tissue was performed,followed by KEGG and GO enrichment analysis.Results Compared with the normal saline group and the aging plasma group,the number of offspring increased in the young plasma group,which reflected higher extrusion rate of first polar body(PB1),decreased fragmentation rate of oocytes and increased conversion rate of two-cell embryos and increased formation rate of blastocysts.There were no significant differences in these indicators between the aging plasma group and the normal saline group.Transcriptomic sequencing revealed that the differentially expressed genes in ovarian tissue of the young plasma group were mainly involved in steroid biosynthesis and metabolic pathways.Among them,the expression level of steroid sulfatase protein was significantly upregulated.Conclusion Systemic infusion of young plasma enhances the reproductive potential of aging ovaries in elderly mice.The sulfated steroid metabolites in plasma may be key substances in restoring ovarian function and delaying the process of ovarian aging.

Objective:To investigate the alterations in voxel-mirrored homotopic connectivity (VMHC) of brain regions, association loop connectivity, and white matter microstructure in patients with white matter hyperintensities (WMH) of presumed vascular origin, and analyze the pathological basis of cognitive impairment in WMH patients.Methods:A prospective study was performed; 75 WMH patients (WMH group) admitted to Jiangsu Shengze Hospital Affiliated to Nanjing Medical University from January 2023 to September 2024 and 67 volunteers without obvious brain diseases (control group) recruited during the same period were enrolled. General data of these participants, and scores of neuropsychological scales such as mini-mental state examination (MMSE), frontal assessment battery (FAB), and trail making test (TMT) were compared between the two groups. Resting-state functional magnetic resonance imaging (rs-fMRI) and diffusion tensor imaging (DTI) data of all participants were collected; rs-fMRI data were then analyzed using VMHC algorithm to calculate and conform the brain regions with significantly different VMHC between the two groups, and these regions were used as seed points to perform functional connectivity with the whole brain; Pearson correlation analyses of VMHC and functional connectivity in these brain regions with scores of neuropsychological scales were performed. DTI data were processed using tract-based spatial statistics (TBSS) method to calculate and conform the brain regions with significantly different diffusion parameters of fiber tracts between the two groups; Pearson correlation analyses of diffusion parameters of the fiber tracts in these brain regions with scores of neuropsychological scales were performed.Results:(1) Comparison of general data and neuropsychological scale scores: proportion of participants with hypertension history was significantly different between the two groups ( P<0.05); scores of TMT-A, TMT-B, and Stroop C scales in the WMH group were significantly higher than those in the control group ( P<0.05). (2) Comparison of VMHC and seed point functional connectivity: compared with that in the control group, the VMHC in bilateral middle occipital gyrus, visual cortex, medial occipitotemporal gyrus, insula, and postcentral gyrus of the WMH group were statistically lower ( P<0.05). Compared with that in the control group, functional connectivity of right visual cortex with right middle temporal gyrus, bilateral precuneus, and right dorsolateral superior frontal gyrus in the WMH group was significantly weakened, and functional connectivity of right postcentral gyrus with right medial occipitotemporal gyrus, left middle temporal gyrus, left visual cortex, and left postcentral gyrus was statistically weakened ( P<0.05). In the WMH group, the VMHC of bilateral insula was negatively correlated with TMT-B score ( r=-0.381, P<0.001), and functional connectivity between right visual cortex and right dorsolateral superior frontal gyrus was negatively correlated with Stroop C score ( r=-0.401, P<0.001). (3) TBSS results: the diffusion parameters of the anterior corona radiata, superior corona radiata, corpus callosum, superior longitudinal fasciculus, and posterior thalamic radiation were statistically significant between the two groups ( P<0.05). In the WMH group, the fractional anisotropy in the genu of the corpus callosum was positively correlated with Stroop C score ( r=0.426, P<0.001), radial diffusivity was negatively correlated with Stroop C score ( r=-0.376, P<0.001), and mean diffusivity of the left anterior corona radiata was negatively correlated with TMT-A score ( r=-0.443, P<0.001). Conclusion:WMH patients have decreased coordination in homotopic brain regions and weakened functional connectivity of association loops, with widely distributed white matter microstructure damages, which may be involved in the neuropathological process of cognitive impairment.

Objective To explore the effect of young plasma intraperitoneal injection on the fertility and ovarian function of aging mice and analyze its potential molecular mechanism.Methods Fifty-four-week-old female C57BL/6 mice and 8-week-old male C57BL/6 mice were selected.Among them,the female mice were randomly divided into three groups:the young plasma group,the aging plasma group and the normal saline group.The young plasma group and the aging plasma group received intraperitoneal injection of plasma from young(25-29 years old)and elderly(45-49 years old)female donors,respectively.Each injection was 500 μL,administered every other day for 2 weeks.The saline group received an equal volume of saline.After the last injection,mating experiments were conducted to evaluate fertility.Ovarian histopathological changes were observed by HE staining.Oocytes and fertilized eggs were collected after superovulation and cultured in vitro to assess oocyte quality and embryo developmental potential.Transcriptomic analysis of ovarian tissue was performed,followed by KEGG and GO enrichment analysis.Results Compared with the normal saline group and the aging plasma group,the number of offspring increased in the young plasma group,which reflected higher extrusion rate of first polar body(PB1),decreased fragmentation rate of oocytes and increased conversion rate of two-cell embryos and increased formation rate of blastocysts.There were no significant differences in these indicators between the aging plasma group and the normal saline group.Transcriptomic sequencing revealed that the differentially expressed genes in ovarian tissue of the young plasma group were mainly involved in steroid biosynthesis and metabolic pathways.Among them,the expression level of steroid sulfatase protein was significantly upregulated.Conclusion Systemic infusion of young plasma enhances the reproductive potential of aging ovaries in elderly mice.The sulfated steroid metabolites in plasma may be key substances in restoring ovarian function and delaying the process of ovarian aging.

Objective To investigate the anti-tumor mechanism of natural compound toosendanin(TSN)combined with olaparib in triple-negative breast cancer(TNBC).Methods Human TNBC cell line MDA-MB-231 was cultured in vitro.Effects of TSN combined with olaparib on autophagy levels and cell viability in MDA-MB-231 cells were evaluated using 0.5,1.0,and 5.0 μmol/L olaparib alone or in combination.Surgical specimens from four TNBC patients who had residual tumors after neoadjuvant chemotherapy were selected to establish patient-derived organoid(PDO)models.The drug sensitivity of TSN combined with olaparib in TNBC patients was detected.Whether TSN combined with olaparib can exert autophagy inhibitory effects and tumor-killing effects in organoid model was verified.Results Olaparib induced autophagy in MDA-MB-231 cell line,and the combination of TSN and olaparib inhibited the proliferation of MDA-MB-231 cells(P＜0.01).In the TNBC PDOs model,the therapeutic effect of olaparib combined with TSN can significantly reduce the proliferation ability of tumor cells compared with olaparib alone.Conclusion The tumor-killing effect of TSN combined with olaparib is superior to that of olaparib alone,and the mechanism may be related to autophagy inhibition.

Objective To investigate the anti-tumor mechanism of natural compound toosendanin(TSN)combined with olaparib in triple-negative breast cancer(TNBC).Methods Human TNBC cell line MDA-MB-231 was cultured in vitro.Effects of TSN combined with olaparib on autophagy levels and cell viability in MDA-MB-231 cells were evaluated using 0.5,1.0,and 5.0 μmol/L olaparib alone or in combination.Surgical specimens from four TNBC patients who had residual tumors after neoadjuvant chemotherapy were selected to establish patient-derived organoid(PDO)models.The drug sensitivity of TSN combined with olaparib in TNBC patients was detected.Whether TSN combined with olaparib can exert autophagy inhibitory effects and tumor-killing effects in organoid model was verified.Results Olaparib induced autophagy in MDA-MB-231 cell line,and the combination of TSN and olaparib inhibited the proliferation of MDA-MB-231 cells(P＜0.01).In the TNBC PDOs model,the therapeutic effect of olaparib combined with TSN can significantly reduce the proliferation ability of tumor cells compared with olaparib alone.Conclusion The tumor-killing effect of TSN combined with olaparib is superior to that of olaparib alone,and the mechanism may be related to autophagy inhibition.

Objective:To investigate the alterations in voxel-mirrored homotopic connectivity (VMHC) of brain regions, association loop connectivity, and white matter microstructure in patients with white matter hyperintensities (WMH) of presumed vascular origin, and analyze the pathological basis of cognitive impairment in WMH patients.Methods:A prospective study was performed; 75 WMH patients (WMH group) admitted to Jiangsu Shengze Hospital Affiliated to Nanjing Medical University from January 2023 to September 2024 and 67 volunteers without obvious brain diseases (control group) recruited during the same period were enrolled. General data of these participants, and scores of neuropsychological scales such as mini-mental state examination (MMSE), frontal assessment battery (FAB), and trail making test (TMT) were compared between the two groups. Resting-state functional magnetic resonance imaging (rs-fMRI) and diffusion tensor imaging (DTI) data of all participants were collected; rs-fMRI data were then analyzed using VMHC algorithm to calculate and conform the brain regions with significantly different VMHC between the two groups, and these regions were used as seed points to perform functional connectivity with the whole brain; Pearson correlation analyses of VMHC and functional connectivity in these brain regions with scores of neuropsychological scales were performed. DTI data were processed using tract-based spatial statistics (TBSS) method to calculate and conform the brain regions with significantly different diffusion parameters of fiber tracts between the two groups; Pearson correlation analyses of diffusion parameters of the fiber tracts in these brain regions with scores of neuropsychological scales were performed.Results:(1) Comparison of general data and neuropsychological scale scores: proportion of participants with hypertension history was significantly different between the two groups ( P<0.05); scores of TMT-A, TMT-B, and Stroop C scales in the WMH group were significantly higher than those in the control group ( P<0.05). (2) Comparison of VMHC and seed point functional connectivity: compared with that in the control group, the VMHC in bilateral middle occipital gyrus, visual cortex, medial occipitotemporal gyrus, insula, and postcentral gyrus of the WMH group were statistically lower ( P<0.05). Compared with that in the control group, functional connectivity of right visual cortex with right middle temporal gyrus, bilateral precuneus, and right dorsolateral superior frontal gyrus in the WMH group was significantly weakened, and functional connectivity of right postcentral gyrus with right medial occipitotemporal gyrus, left middle temporal gyrus, left visual cortex, and left postcentral gyrus was statistically weakened ( P<0.05). In the WMH group, the VMHC of bilateral insula was negatively correlated with TMT-B score ( r=-0.381, P<0.001), and functional connectivity between right visual cortex and right dorsolateral superior frontal gyrus was negatively correlated with Stroop C score ( r=-0.401, P<0.001). (3) TBSS results: the diffusion parameters of the anterior corona radiata, superior corona radiata, corpus callosum, superior longitudinal fasciculus, and posterior thalamic radiation were statistically significant between the two groups ( P<0.05). In the WMH group, the fractional anisotropy in the genu of the corpus callosum was positively correlated with Stroop C score ( r=0.426, P<0.001), radial diffusivity was negatively correlated with Stroop C score ( r=-0.376, P<0.001), and mean diffusivity of the left anterior corona radiata was negatively correlated with TMT-A score ( r=-0.443, P<0.001). Conclusion:WMH patients have decreased coordination in homotopic brain regions and weakened functional connectivity of association loops, with widely distributed white matter microstructure damages, which may be involved in the neuropathological process of cognitive impairment.

Objective:To perform two-step clustering analysis based on urodynamics in patients with neurogenic bladder (NB) and exploring characteristics affecting success rate of sacral neuromodulation (SNM).Methods:The data of 174 NB patients who received SNM therapy in our hospital from September 2018 to August 2023, were retrospectively analysed. There were 94 males and 80 females, with average age of (50.4±17.9)years, median duration of 2 (1, 3)years, and body mass index of 21.5 (20, 25) kg/m 2. Among them, 50 patients had chronic diseases (hypertension or diabetes). Additionally, 77 patients (44.3%) presented with voiding symptoms, 47 patients (27.0%) with storage symptoms, and 50 patients (28.7%) with mixed symptoms. The etiologies included spinal cord lesions in 110 patients (63.2%), brain lesions in 21 patients (12.1%), peripheral nerve lesions in 36 patients (20.7%), and other causes in 7 patients (4.0%). All patients underwent urodynamics and SNM phase 1 test. If patient's subjective symptoms or objective indicators improved > 50%, it is defined as treatment success and feasible to received second stage of SNM. The success rate was analyzed. Clinical indicators and urodynamic parameter were collected for exploring the difference of the above indicators between two groups (with or without conversion). The 174 patients were grouped using two-step clustering analysis, dividing them into two groups. The differences in clinical data, urodynamic parameters, and the conversion rate of SNM phase II between the two groups were compared. Results:Among these patients, 126 (72%) underwent the second stage of SNM. Compared with the non-conversion group, patients in the conversion group exhibited the following characteristics: high proportion of female[51.6% (65/126) vs. 31.2% (15/48)], younger[(48.1± 18.3)year vs. (56.4±15.4) year], lower proportion of history of chronic disease[23.9% (30/126) vs. 41.7% (20/48)], storage predominant symptoms predominant[33.3% (42/126) vs. 10.4% (5/48)], sensitive bladder sensation[50.0% (63/126) vs. 11.1% (14/48)], normal detrusor contractility [26.2% (33/126) vs. 10.4% (5/48)] and detrusor-sphincter dyssynergia [21.4% (27/126) vs. 4.2% (2/48)]and lower post-void residual [105.5(49.3, 231.3) ml vs. 197.9(114.8, 284.8) ml], ( P<0.05). After excluding multicollinearity, patients were divided into two group mainly based on coordination, detrusor stability, and detrusor contractility through two-step clustering analysis: 98 patients in Group 1 and 76 patients in Group 2. The success rate was higher in Group 2 [82.9% (63/76) vs. 64.3% (63/98), P=0.006]. Compared with Group 1, Group 2 exhibited the following characteristics: higher proportion of storage symptoms [43.4% (33/76) vs. 14.3% (14/98)], sensitive bladder sensation [67.1% (51/76) vs. 26.5% (26/98)], detrusor overactivity [46.1% (35/76) vs. 3.1% (3/98)], normal detrusor contractility [43.4% (33/76) vs. 5.1% (5/98)], and detrusor-sphincter dyssynergia [38.2% (29/76) vs. 0] ( P all<0.05). Additionally, group 2 had lower post-void residual [69.0 (23.8, 136.6) ml and 197.9 (123.2, 287.4) ml] and smaller bladder capacity [(281.9±144.9) ml vs. (430.4±176.7) ml] ( P all<0.01). Conclusions:Patients with storage symptoms, low post-void residual, sensitive bladder sensation, detrusor overactivity, normal contractility, or detrusor-sphincter dyssynergia indicate a higher conversion rate to second stage, making them more suitable for SNM testing.