Long-Term Outcome of Photobiomodulation Therapy for Refractory Diabetic Wound After Secretory Carcinoma of the Parotid Gland Surgery: A Case Report.

Objective: To evaluate the efficacy and safety of photobiomodulation therapy (PBMT) in the treatment of diabetic patients with refractory wound. Background: Refractory wound is one of the most challenging clinical complications of diabetes mellitus. Studies have shown that PBMT can promote wound healing in many ways. Methods: We reported a 55-year-old male patient with refractory diabetic wound after secretory carcinoma of the parotid gland surgery responding to 810 nm laser. Results: After PBMT, the refractory diabetic wound healed gradually without adverse events. During follow-up 5-years, the healed wound remained stable and showed no signs of recurrence. Conclusions: PBMT can be potentially considered as a therapeutic method in diabetic patients with refractory diabetic wound.

A Novel Air-Cooled Nd:YAG Laser for the Treatment of the Venous Lakes of the Lips.

Objective: To evaluate the therapeutic effect of a novel air-cooled Nd:YAG laser in the venous lakes of the lips (VLL). Background: The thermal injury is one of the most important issues during laser therapy for venous lakes. Methods: Six pieces of fresh pork livers were used to provide 30 regions with a diameter of 6 mm for experiment in vitro, among which 15 regions were treated by Nd:YAG laser with air cooling until the tissue turned gray-white, whereas the rest were treated without air cooling as control. The operation time of laser irradiation, the degree of temperature increase, and the depth of coagulation tissue were compared between two groups. Then, 60 VLL patients were selected for Nd:YAG laser treatment with or without air cooling. The operation time of laser irradiation, the degree of temperature increase, the postoperative pain visual analog scale (VAS) score, and the percentage of lesions removed within 1 month were compared. Results: In tissue studies, the treated group showed a longer operation time of laser irradiation (p < 0.01), a lower degree of temperature increase (p < 0.01), and there was no significant statistical difference in the depth of coagulation tissue (p = 0.624). In clinical studies, the treated group showed a longer operation time of laser irradiation (p < 0.01), a lower degree of temperature increase (p < 0.01), and a lower VAS score on the 1st and 2nd day, compared with the control group (p < 0.01). Conclusions: Air cooling during Nd:YAG laser for the treatment of VLL can prolong the surgical time, but lowered tissue temperature and reduced patient pain within 2 days under the premise of ensuring the treatment effect.

Transcriptomic evaluation of N6-methyladenosine modification can be used to identify differentially gene and immune-related biological processes in TX mice with liver fibrosis.

BACKGROUND: N6-methyladenosine (m6A) modification controls the stability, splicing, and translation of mRNA, which is important in the development of illnesses. Wilson's disease (WD) is an autosomal recessive liver copper metabolic disorder that causes liver fibrosis. The role of m6A methylation in WD-induced liver fibrosis development is still unclear. Thus, the goal of this study was to examine the scope of m6A methylation and further explore the potential targets related to WD-induced liver fibrosis. RESULTS: A total of 1930 significantly different m6A peaks were found on 1737 mRNAs, of which 993 were hypermethylated and 744 were hypomethylated when comparing normal and WD-induced liver fibrosis mice (n = 3). In parallel, 1261 differentially expressed mRNAs, comprising 557 upregulated and 704 downregulated mRNAs, were found. Overall, 114 mRNAs with significant changes in m6A levels and RNA expression were identified via joint analysis. Then, through PPI network construction and functional enrichment analysis, 12 hub genes were identified, these genes were mainly enriched in the inflammatory response and immunomodulation, and they are associated with immune cell infiltration. CONCLUSIONS: The significant difference in the amount of mRNA m6A modifications indicates that m6A modification is involved in the progression of WD-induced liver fibrosis, and theidentified hub genes are involved in inflammation and immune infiltration. These results may provide insights for subsequent studies on potential regulatory mechanisms.

The accuracy of Fiber-Optic Raman Spectroscopy in the detection and diagnosis of head and neck neoplasm in vivo: a systematic review and meta-analysis.

Purpose: The aim of this article was to review and collectively assess the published studies of fiber-optic Raman spectroscopy (RS) of the in vivo detection and diagnosis of head and neck carcinomas, and to derive a consensus average of the accuracy, sensitivity and specificity. Methods: The authors searched four databases, including Ovid-Medline, Ovid-Embase, Cochrane Library, and the China National Knowledge Infrastructure (CNKI), up to February 2023 for all published studies that assessed the diagnostic accuracy of fiber-optic RS in the in vivo detection of head and neck carcinomas. Nonqualifying studies were screened out in accordance with the specified exclusion criteria, and relevant information about the diagnostic performance of fiber-optic RS was excluded. Publication bias was estimated by Deeks' funnel plot asymmetry test. A random effects model was adopted to calculate the pooled sensitivity, specificity and diagnostic odds ratio (DOR). Additionally, the authors conducted a summary receiver operating characteristic (SROC) curve analysis and threshold analysis, reporting the area under the curve (AUC) to evaluate the overall performance of fiber-optic RS in vivo. Results: Ten studies (including 16 groups of data) were included in this article, and a total of 5365 in vivo Raman spectra (cancer = 1,746; normal = 3,619) were acquired from 877 patients. The pooled sensitivity and specificity of fiber-optic RS of head and neck carcinomas were 0.88 and 0.94, respectively. SROC curves were generated to estimate the overall diagnostic accuracy, and the AUC was 0.96 (95% CI [0.94-0.97]). No significant publication bias was found in this meta-analysis by Deeks' funnel plot asymmetry test. The heterogeneity of these studies was significant; the Q test values of the sensitivity and specificity were 106.23 (P = 0.00) and 64.21 (P = 0.00), respectively, and the I2 index of the sensitivity and specificity were 85.88 (95% CI [79.99-91.77]) and 76.64 (95% CI [65.45-87.83]), respectively. Conclusion: Fiber-optic RS was demonstrated to be a reliable technique for the in vivo detection of head and neck carcinoma with high accuracy. However, considering the high heterogeneity of these studies, more clinical studies are needed to reduce the heterogeneity, and further confirm the utility of fiber-optic Raman spectroscopy in vivo.

Three-Dimensional Gene Regulation Network in Glioblastoma Ferroptosis.

Ferroptosis is an iron-dependent form of cell death, which is reported to be associated with glioma progression and drug sensitivity. Targeting ferroptosis is a potential therapeutic approach for glioma. However, the molecular mechanism of glioma cell ferroptosis is not clear. In this study, we profile the change of 3D chromatin structure in glioblastoma ferroptosis by using HiChIP and study the 3D gene regulation network in glioblastoma ferroptosis. A combination of an analysis of HiChIP and RNA-seq data suggests that change of chromatin loops mediated by 3D chromatin structure regulates gene expressions in glioblastoma ferroptosis. Genes that are regulated by 3D chromatin structures include genes that were reported to function in ferroptosis, like HDM2 and TXNRD1. We propose a new regulatory mechanism governing glioblastoma cell ferroptosis by 3D chromatin structure.

Single-cell RNA sequencing reveals distinct tumor microenvironment of ground glass nodules and solid nodules in lung adenocarcinoma.

Introduction: Lung adenocarcinoma (LUAD) is the most prevalent lung cancer. LUAD presents as ground glass nodules (GGN) and solid nodules (SN) in imaging studies. GGN is an early type of LUAD with good prognosis. However, SN exhibits a more malignant behavior than GGN, including worse pathological staging and tumor prognosis. The mechanism leading to the different malignancy levels of GGN and SN remains elusive. Methods: Three patients with GGN and three patients with SN diagnosed with early LUAD were enrolled. The tumor samples were digested to a single-cell suspension and analyzed using 10× Genomic Single-cell ribonucleic acid sequences (scRNA-seq) techniques. Results: A total of 15,902 cells were obtained and classified into nine major types. The tumor microenvironment (TME) was subsequently described in detail. ScRNA-seq revealed that ribosome-related pathways and cell adhesion played similar but distinct roles in the two groups. SN also had more active cell proliferation, enriched cell cycle regulatory pathways, and severe inflammatory responses. Conclusion: We observed changes in the cellular composition and transcriptomic profile of GGN and SN. The study improved the understanding of the underlying mechanisms of lung carcinogenesis and contributed to lung cancer prevention and treatment.

Efficacy comparisons of solvent-based paclitaxel, liposomal paclitaxel, nanoparticle albumin-bound paclitaxel, and docetaxel after neoadjuvant systemic treatment in breast cancer.

PURPOSE: There are four kinds of taxanes: solvent-based paclitaxel (Sb-P), liposomal paclitaxel (Lps-P), nanoparticle albumin-bound paclitaxel (Nab-P), and docetaxel. This study aims to retrospectively evaluate the efficacy of different taxanes on neoadjuvant systemic treatment (NST) in breast cancer. METHODS: Patients who were diagnosed with breast cancer and had received integral NST from August 2013 to April 2022 were enrolled. The efficacy was divided into total pathological complete response (total-pCR), breast pathological complete response (breast-pCR), and axillary pathological complete response (axillary-pCR) for in-depth analysis and discussion. RESULTS: The choice of taxane was an independent risk factor for total-pCR and breast-pCR rates. The highest total-pCR and breast-pCR rates were found in the Nab-P group. The difference was not significant among all the taxanes in the axillary-pCR rate. CONCLUSION: Nab-P significantly improved the total-pCR and breast-pCR rates. It should be the first choice among taxanes in NST for breast cancer.

Prognostic impact of muscle invasion in buccal mucosa squamous cell carcinoma.

OBJECTIVE: The objective of the study was to assess the prognostic value of muscle invasion (MI), a key histopathological feature of tumor aggressiveness, and construct a superior prognostic prediction model combining the current TNM staging system. MATERIALS AND METHODS: MI was analyzed in the whole-slide images from a total of 301 patients with primary buccal mucosa squamous cell carcinoma (BMSCC). Survival times of patients with/without MI were evaluated by Kaplan-Meier analysis. MI was further combined with the TNM staging system to explore its predictive value for prognosis. Moreover, 204 cases of head and neck carcinoma from the TCGA database were included. RESULTS: MI positive rate reached to 76% (229/301) in patients with BMSCC. MI was associated with poor overall survival (p = 0.012) and disease-free survival (p = 0.022). The novel system (TNM staging combined with MI) revealed strong predictive performance, with the largest area under the curve (OS: p < 0.001, DFS: p < 0.004). MI and the established classification system were also had good predictive ability in the TCGA cohort. CONCLUSIONS: MI is an independent predictor of poor prognosis of BMSCC. The inclusion of MI in prediction system can augment the risk stratification of patients with oral squamous cell carcinoma and may assist in the clinical decision-making process.

Histological evaluation of mouse tongue incisions after Er:YAG laser surgery with different pulse energies versus after conventional scalpel surgery.

PURPOSE: To identify the surgical instrument that allows for optimal healing of tongue incisions. METHODS: An Er:YAG laser was compared with different pulse energies to a conventional scalpel for the incision of mouse tongue tissues. Mice were sacrificed through cervical dislocation at 24, 48, and 72 h postoperatively, followed by extraction of their tongues for incision experiments. The healing of the incisions and expression of inflammation- and pain-related factors in the tongues were compared between the surgical procedure groups. RESULTS: In laser-treated mice, tongue incisions healed the fastest when the laser output energy was 60 MJ per pulse. Macrophage chemotaxis toward the incisional area was triggered on the first postoperative day for the 60-MJ group, while the time for macrophage chemotaxis to the surgical area was later in the 80-MJ group. Tumor necrosis factor-alpha expression increased and then decreased in the 80-MJ group; however, it gradually decreased in the 60-MJ and conventional scalpel groups. Prostaglandin E2 expression increased and then decreased in the 80-MJ and conventional scalpel groups but gradually decreased in the 60-MJ group. The expression of transforming growth factor beta 1 gradually decreased in the 60-MJ and 80-MJ groups but gradually increased in the conventional scalpel group. CONCLUSION: Compared with surgical procedures using conventional scalpels, those using an Er:YAG laser with appropriate pulse energies can inhibit inflammation in the incisional area and promote incision healing. The use of an Er:YAG laser with appropriate pulse energies can alleviate intraoperative and postoperative pain in the incisional area.

Oral tongue squamous cell carcinoma diagnosis from tissue metabolic profiling.

OBJECTIVE: Disease metabolomes have been studied for identifying diagnostic and predictive biomarkers of pathology. Oral tongue squamous cell carcinoma (OTSCC) is one of the most prevalent subtypes of head and neck squamous cell carcinoma, yet the profile and diagnostic value of its tissue metabolite are unclear. SUBJECTS AND METHODS: Tumor tissue samples and matched normal mucosal tissue samples were collected from 40 OTSCC patients. Untargeted metabolic analysis by liquid chromatography-mass spectrometry/mass spectrometry, in positive and negative ion modes, was used to identify dysregulated metabolites in OTSCC. Further, utilizing LASSO regression and receiver operating characteristic analyses, biomarker metabolites were selected and validated, and a diagnostic model was established. RESULTS: One hundred and ninety metabolites were detected. The OTSCC had a total of 89 dysregulated metabolites, of which 73 were elevated. A diagnostic panel of nine metabolites was subsequently created that could accurately identify OTSCC with 100% sensitivity of 100%, 100% specificity and an AUC of 1.00. CONCLUSIONS: This study identified distinct metabolic characteristics of OTSCC and established a diagnostic model. Our research also contributes to the investigation of the pathogenesis of OTSCC.

Generation of gastirc insulin-secreting organoids from human stomach sample.

Stomach stem cells are accessible by biopsy and propagate robustly in culture, offering an invaluable resource for autologous cell therapies. Here we describe a detailed protocol to isolate, expand, engineer and differentiate human gastric stem cells (hGSCs) into pancreatic islet-like organoids containing abundant gastric insulin-secreting (GINS) cells that resemble beta-cells in molecular hallmarks and function. Sequential activation of the inducing factors NGN3 and PDX1-MAFA led hGSCs onto a novel differentiation path, including endocrine progenitor and GINS precursor, before adopting beta-cell identity, at efficiencies close to 70%. GINS organoids acquired glucose-stimulated insulin secretion in 10 days post differentiation.

Multiperspective quantitative tumor-stroma ratio reveals histological areas associated with poor outcomes in oral squamous cell carcinoma.

AIMS: Different regions of oral squamous cell carcinoma (OSCC) have particular histopathological characteristics, and the individual histological characteristics of the tumors are poorly understood. Therefore, calculating the proportion of tumor cells in different regions that allow assessment of the prognostic outcomes for OSCC patients would be of great clinical significance. METHODS AND RESULTS: We established an open-source software-based analytic pipeline that defines the inner tumor and invasive tumor front (ITF) in pancytokeratin-stained whole slide images (WSIs) and quantifies the tumor-stroma ratio (TSR) within the two regions. We applied this method to 114 patients with OSCC and predicted patient prognosis by the TSR. The proportion of tumor area in the inner tumor was generally higher than that in the ITF (p < 0.0001). TSR was an independent prognostic factor for overall survival (OS) (p = 0.016), disease-free survival (DFS) (p = 0.026), and relapse-free survival (RFS) (p = 0.037) in inner tumor, and TSR was an independent prognostic factor for OS (p = 0.00052), DFS (p = 0.035), and metastasis-free survival (MFS) (p = 0.038) in the ITF. Tumor-low status was associated with poorer prognosis. There was a significant correlation between the TSR and perineural invasion (PNI) in the inner tumor (p = 0.009). CONCLUSIONS: The histopathological characteristics of different regions of OSCC may be used to develop the potential prognostic markers. The TSR of the inner tumor is more targeted in predicting prognosis and accurately assesses the risk of PNI+.

Osteogenic Differentiation Effect of Human Periodontal Ligament Stem-Cell Initial Cell Density on Autologous Cells and Human Bone Marrow Stromal Cells.

Stem cells have differentiation and regulation functions. Here, we discussed the impact of cell culture density on stem cell proliferation, osteoblastogenesis, and regulation. To discuss the effect of the initial culture density of human periodontal ligament stem cells (hPDLSCs) on the osteogenic differentiation of autologous cells, we found that the hPDLSC proliferation rate decreased with an increase in the initial plating density (0.5-8 × 104 cells/cm2) for the 48 h culture cycle. After hPDLSCs induced osteogenic differentiation for 14 days with different initial cell culture densities, the expression of osteoprotegerin (OPG) and runt-related transcription factor 2(RUNX2) and the OPG/ Receptor Activator of Nuclear Factor-κ B Ligand (RANKL) ratio were the highest in the hPDLSCs initially plated at a density of 2 × 104 cells/cm2, and the average cell calcium concentration was also the highest. To study hPDLSCs regulating the osteoblastic differentiation of other cells, we used 50 µg/mL of secreted exosomes derived from hPDLSCs cultured using different initial cell densities to induce human bone marrow stromal cell (hBMSC) osteogenesis. After 14 days, the results indicated that the gene expression of OPG, Osteocalcin(OCN,)RUNX2, and osterix and the OPG/RANKL ratio were the highest in the 2 × 104 cells/cm2 initial cell density group, and the average calcium concentration was also the highest. This provides a new idea for the clinical application of stem cell osteogenesis.

Stomach-derived human insulin-secreting organoids restore glucose homeostasis.

Gut stem cells are accessible by biopsy and propagate robustly in culture, offering an invaluable resource for autologous cell therapies. Insulin-producing cells can be induced in mouse gut, but it has not been possible to generate abundant and durable insulin-secreting cells from human gut tissues to evaluate their potential as a cell therapy for diabetes. Here we describe a protocol to differentiate cultured human gastric stem cells into pancreatic islet-like organoids containing gastric insulin-secreting (GINS) cells that resemble ß-cells in molecular hallmarks and function. Sequential activation of the inducing factors NGN3 and PDX1-MAFA led human gastric stem cells onto a distinctive differentiation path, including a SOX4High endocrine and GalaninHigh GINS precursor, before adopting ß-cell identity, at efficiencies close to 70%. GINS organoids acquired glucose-stimulated insulin secretion in 10 days and restored glucose homeostasis for over 100 days in diabetic mice after transplantation, providing proof of concept for a promising approach to treat diabetes.

Clinical and imaging features of eight cases of Ewing sarcoma of the jaw.

OBJECTIVES: This study investigate the clinical and imaging features of Ewing sarcoma (ES) of the jaw. METHODS: Eight cases of pathologically diagnosed ES of the jaw from January 2010 to June 2022 were included in the study. Clinical and radiological features were retrospectively analyzed. RESULTS: Among the eight cases, the mean age at onset was 29.4 years, and the male to female ratio was 7∶1. The predilecting site was the posterior part of mandible, accounting for 75% of the cases. The lesions often exhibited early numbness of the lower lip and lymphadenopathy. The main radiographic manifestation of mandibular lesions was ill-defined radiolucency, mixed with fibrous or brush-like tumor matrix, and soft tissue mass. The maxillary ES lesions mainly presented as lytic bone destruction accompanied by adjacent soft tissue mass. Periosteal ossification was rarely seen. CONCLUSIONS: The clinical and imaging characteristics of ES in the jaw are helpful for its diagnosis.

Regulation of TREM2 expression by transcription factor YY1 and its protective effect against Alzheimer's disease.

TREM2 encoding the transmembrane receptor protein TREM2 is a risk gene of Alzheimer's disease (AD), and the impairment of TREM2 functions in microglia due to mutations in TREM2 may significantly increase the risk of AD by promoting AD pathologies. However, how the expression of TREM2 is regulated and the transcription factors required for TREM2 expression are largely unknown. By luciferase assay, DNA pull-down, and in silico predictions, we identified Yin Yang 1(YY1) as a binding protein of the minimal promoter of the TREM2 gene, and the binding was further confirmed by EMSA and DNA pull-down assay. shRNA-mediated YY1 silencing significantly reduced the activity of the TREM2 minimal promoter and TREM2 protein levels in the microglial cell line BV2 and the neuroblastoma Neuro2A. Furthermore, we found that the levels of TREM2 and YY1 were both downregulated in lipopolysaccharide-treated BV2 cells and in the brain of AD model mice. These results demonstrated that YY1 plays a crucial role in the regulation of TREM2 expression. Our study suggests that microglial YY1 could be targeted to maintain TREM2 expression for AD prevention and therapy.

Investigation of periodontal status and bacterial composition aroundmini-implants.

INTRODUCTION: Mini-implants are now widely used in orthodontic treatment. Soft-tissue inflammation around the mini-implant is an important factor affecting its stability. This study aimed to investigate the periodontal status and the bacterial composition around mini-implants. METHODS: A total of 79 mini-implants in 40 patients (aged 18-45 years) were evaluated in this study. The mini-implant probing depth (mPD), mini-implant gingival sulcus bleeding index (mBI), mini-implant plaque index (mPLI), and the composition of the supragingival and subgingival plaque around the mini-implants were recorded. After Congo red staining, the bacteria were classified and counted under a light microscope. RESULTS: The mPLI and mBI around mini-implants in the infrazygomatic crest were higher than those in the buccal shelf and interradicular area. The mPD was higher on the coronal site of the mini-implant than on the mesial, distal, and apical sites in the infrazygomatic crest. The mPLI around the mini-implant was positively correlated with the mBI, and the mBI was positively correlated with the mPD. The supragingival and subgingival bacterial composition around the mini-implants was similar to that of natural teeth. Compared with supragingival bacterial composition, the subgingival bacteria of mini-implants had a significantly lower proportion of cocci and a higher proportion of bacilli and spirochetes. CONCLUSIONS: The bacteria composition of the plaque and the location are important factors in the inflammation around mini-implants. Similar to natural teeth, mini-implants require health maintenance to prevent inflammation of the surrounding soft tissue and maintain stability.

Evolution characteristic of atmospheric black carbon particles at a coastal site in the Pearl River Delta, China.

The mixing of black carbon (BC) with secondary materials is a major uncertainty source in assessing its radiative forcing. However, current understanding of the formation and evolution of various BC components is limited, particularly in the Pearl River Delta, China. This study measured submicron BC-associated nonrefractory materials and the total submicron nonrefractory materials using a soot particle aerosol mass spectrometer and a high-resolution time-of-flight aerosol mass spectrometer, respectively, at a coastal site in Shenzhen, China. Two distinct atmospheric conditions were also identified to further explore the distinctive evolution of BC-associated components: polluted period (PP) and clean period (CP). Comparing the components of two particles, we found that more-oxidized organic factor (MO-OOA) prefers to form on BC during PP rather CP. The formation of MO-OOA on BC (MO-OOABC) was affected by both enhanced photochemical processes and nocturnal heterogeneous processes. Enhanced photo-reactivity of BC, photochemistry during the daytime, and heterogeneous reaction at nighttime were potential pathways for MO-OOABC formation during PP. The fresh BC surface was favorable for the formation of MO-OOABC. Our study shows the evolution of BC-associated components under different atmospheric conditions, which should be considered in regional climate models to improve the assessment of the climate effects of BC.

Nanoparticle albumin-bound paclitaxel-based neoadjuvant regimen: A promising treatment option for HER2-low-positive breast cancer.

This study aimed to compare the efficacy of neoadjuvant systemic therapy (NST) with solvent-based paclitaxel (Sb-P), liposomal paclitaxel (Lps-P), nanoparticle albumin-bound paclitaxel (Nab-P), and docetaxel in human epidermal growth factor receptor 2 (HER2)-low-positive and HER2-zero breast cancers. A total of 430 patients receiving 2-weekly dose-dense epirubicin and cyclophosphamide (EC) followed by 2-weekly paclitaxel (Sb-P, Lps-P, or Nab-P), or 3-weekly EC followed by 3-weekly docetaxel for NST were enrolled in the study. In HER2-low-positive patients, the pathological complete response (pCR) rate in Nab-P group was significantly higher than that in the other three paclitaxel groups (2.8 % in Sb-P group, 4.7 % in Lps-P group, 23.2 % in Nab-P group and 3.2 % in docetaxel group, p < 0.001). In HER2-zero patients, the pCR rate did not differ significantly among the four paclitaxel groups (p = 0.278). The NST regimen containing Nab-P could be considered a promising treatment option in HER2-low-positive breast cancer.

Prognostic analysis of three forms of Ki-67 in patients with breast cancer with non-pathological complete response before and after neoadjuvant systemic treatment.

BACKGROUND: Patients who do not achieve a pathological complete response (pCR) after neoadjuvant systemic treatment (NST) have a significantly worse prognosis. A reliable predictor of prognosis is required to further subdivide non-pCR patients. To date, the prognostic role in terms of disease-free survival (DFS) between the terminal index of Ki-67 after surgery (Ki-67T ) and the combination of the baseline Ki-67 at biopsy before NST (Ki-67B ) and the percentage change in Ki-67 before and after NST (Ki-67C ) has not been compared. AIM: This study aimed to explore the most useful form or combination of Ki-67 that can provide prognostic information to non-pCR patients. PATIENTS AND METHODS: We retrospectively reviewed 499 patients who were diagnosed with inoperable breast cancer between August 2013 and December 2020 and received NST with anthracycline plus taxane. RESULTS: Among all the patients, 335 did not achieve pCR (with a follow-up period of ≥1 year). The median follow-up duration was 36 months. The optimal cutoff value of Ki-67C to predict a DFS was 30%. A significantly worse DFS was observed in patients with a low Ki-67C (p < 0.001). In addition, the exploratory subgroup analysis showed relatively good internal consistency. Ki-67C and Ki-67T were considered as independent risk factors for DFS (both p < 0.001). The forecasting model combining Ki-67B and Ki-67C showed a significantly higher area under the curve at years 3 and 5 than Ki-67T (p = 0.029 and p = 0.022, respectively). CONCLUSIONS: Ki-67C and Ki-67T were good independent predictors of DFS, whereas Ki-67B was a slightly inferior predictor. The combination of Ki-67B and Ki-67C is superior to Ki-67T for predicting DFS, especially at longer follow-ups. Regarding clinical application, this combination could be used as a novel indicator for predicting DFS to more clearly identify high-risk patients.