Objective To investigate the heterogeneity and key molecular features of epithelial cells in triple-negative breast cancer (TNBC), identify prognostic biomarkers, and develop a robust survival prediction model. Methods Using TNBC single-cell transcriptomic data, epithelial cells were extracted, normalized, and subclustered to characterize their molecular signatures and functional differences. High-dimensional weighted gene co-expression network analysis (hdWGCNA) was applied to establish co-expression modules in epithelial cells. Multiple machine learning algorithms were integrated to select key prognostic genes and develop a risk-score model, whose performance was evaluated using receiver operating characteristic (ROC) curves and Kaplan-Meier (K-M) survival analysis. In addition, the immune microenvironment features and potential drug-response differences between the high- and low-risk groups were systematically assessed. Finally, PCR was performed to validate the expression differences of the key genes between tumor and normal tissues. Results We characterized the composition and molecular features of TNBC epithelial subpopulations and identified a TNBC-associated epithelial subset. By integrating hdWGCNA with machine learning approaches, 10 key genes were selected to construct a prognostic model, which effectively stratified patients into distinct survival-risk groups and demonstrated favorable predictive performance in ROC and K-M analyses. Immune profiling revealed the differences in the infiltration levels of seven immune cell types and immune function-related features between the high- and low-risk groups. Drug-sensitivity analysis suggested potential differential responses to eight agents across the risk groups. PCR validation further confirmed the differential expression of the ten signature genes between tumor and normal tissues. Conclusion This study reveals epithelial heterogeneity in TNBC at single-cell resolution and establishes a 10-gene prognostic model, which may facilitate the stratification of TNBC risk and the evaluation of immune characteristics and potential therapeutic strategies.

BACKGROUND:Human periodontal ligament stem cells are potential functional cells for periodontal tissue engineering.However,long-term in vitro culture may lead to reduced stemness and replicative senescence of periodontal ligament stem cells,which may impair the therapeutic effect of human periodontal ligament stem cells. OBJECTIVE:To investigate the effect of oxymatrine on the stemness maintenance and osteogenic differentiation of periodontal ligament stem cells in vitro,and to explore the potential mechanism. METHODS:Periodontal ligament stem cells were isolated from human periodontal ligament tissues by tissue explant enzyme digestion and cultured.The surface markers of mesenchymal cells were identified by flow cytometry.Periodontal ligament stem cells were incubated with 0,2.5,5,and 10 μg/mL oxymatrine.The effect of oxymatrine on the proliferation activity of periodontal ligament stem cells was detected by CCK8 assay.The appropriate drug concentration for subsequent experiments was screened.Western blot assay was used to detect the expression of stem cell non-specific proteins SOX2 and OCT4 in periodontal ligament stem cells.qRT-PCR and western blot assay were used to detect the expression levels of related osteogenic genes and proteins in periodontal ligament stem cells. RESULTS AND CONCLUSION:(1)The results of CCK8 assay showed that 2.5 μg/mL oxymatrine significantly enhanced the proliferative activity of periodontal stem cells,and the subsequent experiment selected 2.5 μg/mL oxymatrine to intervene.(2)Compared with the blank control group,the protein expression level of SOX2,a stem marker of periodontal ligament stem cells in the oxymatrine group did not change significantly(P>0.05),and the expression of OCT4 was significantly up-regulated(P<0.05).(3)Compared with the osteogenic induction group,the osteogenic genes ALP,RUNX2 mRNA expression and their osteogenic associated protein ALP protein expression of periodontal ligament stem cells were significantly down-regulated in the oxymatrine+osteogenic induction group(P<0.05).(4)The oxymatrine up-regulated the expression of stemness markers of periodontal ligament stem cells and inhibited the bone differentiation of periodontal ligament stem cells,and the results of high-throughput sequencing showed that it may be associated with WNT2,WNT16,COMP,and BMP6.

BACKGROUND:Flap transplantation technique is a commonly used surgical procedure for the treatment of severe tissue defects,but postoperative flap necrosis is easily triggered by ischemia-reperfusion injury.Therefore,it is still an important research topic to improve the survival rate of transplanted flaps. OBJECTIVE:To review the pathogenesis and latest treatment progress of flap ischemia-reperfusion injury. METHODS:CNKI,WanFang Database and PubMed database were searched for relevant literature published from 2014 to 2024.The search terms used were"flap,ischemia-reperfusion injury,inflammatory response,oxidative stress,Ca2+overload,apoptosis,mesenchymal stem cells,platelet-rich plasma,signaling pathways,shock wave,pretreatment"in Chinese and English.After elimination of irrelevant literature,poor quality and obsolete literature,77 documents were finally included for review. RESULTS AND CONCLUSION:Flap ischemia/reperfusion injury may be related to pathological factors such as inflammatory response,oxidative stress response,Ca2+overload,and apoptosis,which can cause apoptosis of vascular endothelial cells,vascular damage and microcirculation disorders in the flap,and eventually lead to flap necrosis.Studies have found that mesenchymal stem cell transplantation,platelet-rich plasma,signaling pathway modulators,shock waves,and pretreatment can alleviate flap ischemia/reperfusion injuries from different aspects and to varying degrees,and reduce the necrosis rate and necrosis area of the grafted flap.Although there are many therapeutic methods for skin flap ischemia/reperfusion injury,a unified and effective therapeutic method has not yet been developed in the clinic,and the advantages and disadvantages of various therapeutic methods have not yet been compared.Most of the studies remain in the stage of animal experiments,rarely involving clinical observations.Therefore,a lot of research is required in the future to gradually move from animal experiments to the clinic in order to better serve the clinic.

Flap surgery is a complex surgical procedure that has become an effective method for the treatment of many diseases and traumas.Flap survival is closely related to a variety of factors including cellular autophagy,oxidative stress,inflammatory response,mesenchymal stem cells(MSCs)function,and vascular regeneration.Cellular autophagy maintains intracellular homeostasis and plays a key role in reducing oxidative stress and inflammation and promoting injury repair.Excessive oxidative stress and inflammatory responses pose a threat to flaps,affecting their survival and successful transplantation.Endothelial cells are involved in vascu-lar regeneration through proliferation,migration,and production of angiogenic factors,and vascular endothelial growth factor directly promotes blood vessel formation and maintains endothelial cell function.MSCs play an important role in promoting flap survival and tissue repair due to their unique biological properties and multiple mechanisms of action.The multiple roles played by cellular autophagy,oxidative stress,inflammatory response,MSCs function,and vascular regeneration in influencing postoperative flap survival are hereby elaborated.The aim is to provide a basis for the clinical application of regulating the above factors to improve postoperative flap survival,improve the success rate of flap surgery,reduce complications,and bring more hope for the recovery and quality of life of patients.

Objective:To explore the diagnostic value of fractional flow reserve (FFR) based on optical coherence tomography (OCT) for coronary functional ischemia, and to investigate the feasibility of synchronously obtaining Doppler blood flow velocity information with the help of OCT technology.Methods:This study was a single-center, prospective, self-controlled clinical study on coronary heart disease patients who underwent OCT and FFR assessment at Department of Cardiology, the Second Hospital of Jilin University from January 2024 to February 2025. Linear regression analysis was used to evaluate the correlation and consistency between OCT-FFR and FFR. With FFR≤0.80 as the gold standard for judging whether the target vessel was ischemic, the diagnostic performance of OCT-FFR was evaluated, and the diagnostic value of OCT-FFR for significant coronary artery ischemia was assessed using subject operating characteristic curves. Blood flow OCT data were analyzed by Doppler processing algorithm to obtain information on coronary blood flow velocity.Results:A total of 31 vessels from 28 patients were analyzed, the correlation coefficient r between OCT-FFR and FFR of the 31 vessels was 0.84 ( P<0.001). With FFR as the gold standard to determine whether a coronary artery is ischemic, accuracy, sensitivity, specificity, positive predictive value, and negative predictive value for OCT-FFR was 93.55%, 75.00%, 100.00%, 1.00, 0.92. In clinical practice, Doppler OCT was used for the first time to obtain information on coronary blood flow velocity. Conclusion:OCT-FFR had an excellent correlation and consistency with FFR in judging whether there is coronary artery ischemia. The feasibility of the simultaneous acquisition of OCT-FFR and Doppler flow velocity information in coronary stenosis was preliminarily verified.

Objective:To investigate the clinicopathological characteristics of type 2 autoimmune pancreatitis (AIP) and to explore its relationship with inflammatory bowel disease (IBD).Methods:AIP cases confirmed by pathology in the First Affiliated Hospital of Zhejiang University School of Medicine from 2009 to 2024 were collected. According to the International Consensus Diagnostic Criteria (ICDC) for AIP, 11 patients were identified as histological level 1 (definite) or level 2 (probable) type 2 AIP. Their clinical manifestations, laboratory test results, imaging features, and histopathological characteristics were analyzed, and a follow-up was conducted. Meanwhile, 130 patients with type 1 AIP diagnosed in our hospital during the same period were selected as control group.Results:Among 141 AIP patients, 11 cases (7.8%, 11/141) were diagnosed with type 2 AIP, including 7 cases of histologically level 1 and 4 cases of level 2. There were 10 male patients and 1 female patient, with a median age of 37(31,46) years (range: 25-47 years). Three patients were complicated with ulcerative colitis (UC). Compared with type 1 AIP patients, type 2 AIP patients were younger, often presented with acute pancreatitis or abdominal pain as the initial symptom, and had a close association with IBD ( P<0.05). Laboratory tests showed that only 1 patient had slightly elevated serum IgG4, while the other 10 patients had normal serum IgG4 levels. Serum CA19-9 was elevated in 8 patients, and the percentage of peripheral blood neutrophils was increased in 9 patients. Imaging findings revealed diffuse pancreatic enlargement in 8 patients and localized enlargement in 3 patients (2 cases in the pancreatic head and 1 case in the pancreatic body-tail). Magnetic resonance cholangiopancreatography (MRCP) showed main pancreatic duct stenosis in 5 cases (5/7). Histopathological features included 7 cases of level 1 type 2 AIP that showed neutrophilic infiltration in the pancreatic duct epithelium and massive neutrophilic infiltration between the acini. Immunohistochemistry showed that only 1 case had <5 IgG4-positive plasma cells per high-power field (HPF), while the other 10 cases were negative. All 11 patients with type 2 AIP received steroid therapy, and no recurrence was observed during the follow-up period of 5 to 174 months. Conclusions:Type 2 AIP has unique clinicopathological characteristics. It is more commonly found in young patients and often presents with manifestations similar to acute pancreatitis. Histologically, neutrophilic infiltration in the ductal epithelium is the common feature. Type 2 AIP is closely associated with IBD, especially UC.

Purpose To evaluate the value of tumors invasion in the central airway(TICA)in predicting the severe immune checkpoint inhibitor-related pneumonitis(S-CIP)in lung cancer patients using propensity score matching(PSM).Materials and Methods The intact data of 162 consecutive lung cancer patients who received treatment with immune checkpoint inhibitors in Xi'an International Medical Center Hospital from September 2019 to March 2022 were retrospectively collected.Patients were divided into S-CIP group(23 cases)and non-S-CIP group(139 cases)according to the presence of S-CIP.The demographic information of the patients,including gender,age,history of smoking,thoracic radiotherapy histology,baseline lung diseases,classification,TNM stage,tumor location as well as TICA were collected.A binary Logistic regression was used to analyze the confounding factors and independent risk factors of S-CIP and to predict the development of S-CIP.A 1:1 matching was performed by the nearest neighbor method for PSM.The PSM was used to pair the two groups,and the value of TICA in predicting S-CIP before and after PSM was compared.The receiver operating characteristic curve and the area under the curve were used for model performance based on TICA.Results Before PSM,the proportion of baseline lung diseases(78.3%vs.32.4%,OR=6.802,P=0.001),thoracic radiotherapy history(69.6%vs.30.2%,OR=5.300,P=0.002)and TICA(65.2%vs.27.3%,OR=5.882,P=0.001)in the S-CIP group was higher than those in the non-S-CIP group,and were independent risk factor for predicting S-CIP.After PSM,20 patients were included in each group.The presence of TICA was higher in S-CIP group than that in the non-S-CIP group(60.0%vs.20.0%,OR=6.000,P=0.013).The area under the curves of Logistic regression model based on TICA was 0.700(95%CI 0.534-0.866).Conclusion TICA is an independent risk factor for development of S-CIP,which has moderate degree of accuracy in predicting S-CIP,can be used for risk prediction and early intervention to reduce the poor prognosis of S-CIP patients.

Objective:To analyze and compare the interfractional setup errors between two body posi-tioning fixation methods(lithotomy position with carbon fiber full-body fixation frame vs.conventional carbon fiber body fixation frame combined with thermoplastic membrane)in radical radiotherapy for pros-tate cancer,and to calculate the clinical target volume(CTV)to planning target volume(PTV)margin(MPTV)for both methods to optimize immobilization techniques and radiotherapy workflows.Methods:A retrospective analysis was conducted on 37 consecutive patients who underwent radical prostate radio-therapy at Peking University First Hospital between August 2021 and March 2023.The patients were di-vided into two groups based on the immobilization method:Group A(18 patients,450 CBCT image sets)used a carbon fiber whole-body fixator in the lithotomy position,while Group B(19 patients,461 CBCT image sets)used a conventional carbon fiber fixator combined with a thermoplastic mask.All the patients underwent daily cone-beam computed tomography(CBCT)image guidance.Bone registration combined with manual registration was used to obtain the setup error data in the left-right(X),cranio-caudal(Y)and anterior-posterior(Z)directions.The positioning errors of the two groups were compared by using the independent sample t-test,the Mann-Whitney U test and the chi-square test.The average positioning error,systematic positioning error(Σ)and random positioning error(δ)were calculated,and the CTV-PTV extension distance was calculated by using the(MPTV=2.5Σ+0.7δ).Results:The analysis of the setup errors in the three-dimensional direction showed significant differences between the two groups(all P＜0.01).Specifically,the median(quartile)absolute values of the errors in the X,Y,and Z di-rections of group A were[0.40(0.20,0.70)cm,0.50(0.30,0.80)cm,and 0.35(0.20,0.60)cm],respectively.In group B,the corresponding values were significantly reduced to[0.20(0.10,0.40)cm,0.40(0.20,0.70)cm and 0.20(0.10,0.40)cm].The results of Mann-Whitney U test showed that the differences in each direction were highly statistically significant(X:z=-6.86;Y:z=-2.76;Z:z=-5.71).The cumulative distribution ratio of the setup error displacement within 0.5 cm in the X,Y,and Z directions in group A and group B were 297(66.0％)and 408(88.5％)(P＜0.01),250(55.6％)and 285(61.8％)(P=0.055),308(68.4％)and 391(84.8％)(P＜0.01),re-spectively.The CTV-PTV margins in three directions were X 0.66 cm in group A and 0.35 cm in group B;Y0.67 cm and 0.45 cm;Z 0.54 cm and 0.42 cm.Conclusion:Conventional carbon fiber human body fixator combined with thermoplastic film can significantly reduce the setup error.However,the car-bon fiber whole-body fixator in the lithotomy position demonstrated larger setup errors in the X,Y,and Z directions compared with the conventional fixator combined with a thermoplastic mask,indicating the need for further workflow optimization.Given that the lithotomy position is essential for transperineal ultrasound-guided techniques,further research and improvements are required to achieve higher positioning accuracy.

Objective:To compare the efficacy of robot system-assisted versus freehand screw revision for ankylosing spondylitis (AS) with lower cervical fractures.Methods:A multicenter retrospective cohort study was conducted to analyze the clinical data of 57 patients with AS combined with lower cervical fractures admitted to Honghui Hospital Affiliated to Xi'an Jiaotong University School of Medicine, Henan Provincial People's Hospital, Zhengzhou Orthopedic Hospital, and Qilu Hospital of Shandong University, including 46 males and 11 females, aged 38-77 years [(65.4±9.5)years]. Injury segments involved C 3 in 7 patients, C 4 in 13, C 5 in 25, C 6 in 10, and C 7 in 2. All the patients underwent revision surgery, among whom, 22 patients were treated with robot system-assisted cervical pedicle screw placement (robot nailing group, with 190 screws), and 35 with freehand cervical pedicle screw placement (freehand nailing group, with 300 screws). The operative duration, intraoperative bleeding volume, frequency of intraoperative fluoroscopy, incision length, and length of hospital stay of the two groups were compared; the time of single nscrew insertion, the number of single nail revisions, the distance between screws and the anterior cortex, the accuracy of screw placement of grade 0 and grade 0+1 were recorded in the two groups. The visual analogue scale (VAS), Japanese Orthopedic Society (JOA) score, neck dysfunction index (NDI), American Spine Injury Association (ASIA) classification before operation, at 3 days, 3 months after operation and at the last follow-up were compared between the two groups. The complication rate was also noted. Results:All the patients were followed up for 12-16 months [(14.3±2.1)months]. The operative duration, intraoperative bleeding volume, and frequency of intraoperative fluoroscopy were (186.4±12.9)minutes, (486.1±68.6)ml, and (3.4±1.3)times in the robot nailing group, which were shorter or less than (206.7±14.4)minutes, (660.3±45.2)ml, and (13.5±3.6)times in the freehand nailing group ( P<0.01). The incision length was (9.4±2.4)cm in the robot nailing group, longer than (5.6±1.2)cm in the freehand nailing group ( P<0.01), and the length of hospital stay was (3.7±0.4)days, shorter than (4.4±1.4)days in the freehand nailing group ( P<0.01). The length of single nail insertion, the number of single nail revision, and the distance between the screws and the front cortex were (6.5±0.4)minutes, (1.1±0.1)times, and (3.5±1.3)mm in the robot nailing group, which were shorter or less than (11.6±0.2)minutes, (1.5±0.2)times, and (12.4±4.7)mm in the freehand nailing group ( P<0.01). The accuracy of the screw placement in the robot nailing group was 90.0% (171/190) and 95.8% (182/190) with level 0 and 0+1 screws, better than 80.0% (240/300) and 89.0% (267/300) in the freehand nailing group ( P<0.05). There was no significant difference in VAS, JOA score, NDI, or ASIA grading between the two groups before operation ( P>0.05). The VAS, JOA, and NDI scores at 3 days after operation were (3.1±0.6)points, (12.1±1.2)points, and (15.6±2.9)points, respectively in the robot nailing group, which were better than (5.0±1.4)points, (11.3±1.1)points and (22.5±3.7)points, respectively in the freehand nailing group ( P<0.05). No statistically significant difference was observed in the ASIA grade between the two groups at 3 days after operation ( P>0.05). There were no significant differences in VAS, JOA, NDI scores, or ASIA grading between the two groups at 3 months after operation and at the last follow-up ( P>0.05). Compared with those before operation, the VAS, JOA, NDI scores, and ASIA grading were significantly improved at 3 days, 3 months after operation and at the last follow-up in the two groups, which were further improved with the passage of time. Two patients in the robot nailing group had pneumonia, with a complication rate of 9% (2/22), while 2 patients in the freehand nailing group had dural sac rupture and cerebrospinal fluid leakage and 3 had lung infection after operation, with a complication rate of 14% (5/35) ( P<0.05). Conclusion:Compared with freehand nailing, the robot system-assisted nailing revision for AS with lower cervical fracture has more advantages in terms of the operative duration, length of hospital stay, intraoperative bleeding volume, frequency of intraoperative fluoroscopy nailing speed and accuracy, screw holding force, early pain relief, function restoration, and complication rate, despite longer surgical incision.

As a common ophthalmic disease, pterygium exhibits a considerably high incidence in global area, particularly in regions with harsh natural environment. While it is unlikely to cause blindness, pterygium frequently disrupts the normal homeostasis of the tear film, leading to discomfort such as dry eyes and foreign body sensation, thereby significantly impairing patients' quality of life. Furthermore, the high surgical intervention rate for pterygium consumes substantial medical resources. Therefore, investigating the underlying mechanisms of pterygium development is essential for its prevention and control. Extensive research has demonstrated the correlation between solar radiation and pterygium occurrence, confirming that ultraviolet rays within solar radiation contribute to the onset and progression of pterygium through multiple pathways, including direct DNA damage to ocular surface cells, oxidative stress-induced injury, involvement in inflammatory regulation, and extracellular matrix remodeling. This paper reviews both domestic and international epidemiological trends of pterygium, as well as the multifaceted mechanisms by which solar radiation influences pterygium development. The aim is to enhance understanding of the relationship between solar exposure and pterygium occurrence, while emphasizing and guiding public awareness of solar radiation protection in clinical practice.