Triple-negative breast cancer (TNBC) represents a distinctive subtype, characterized by the absence of estrogen receptors, progesterone receptors, and human epidermal growth factor receptor 2 (HER2). Due to its high inter-tumor and intra-tumor heterogeneity, TNBC poses significant chanllenges for personalized diagnosis and treatment. The advant of clustered regular interspaced short palindromic repeats (CRISPR) technology has profoundly enhanced our understanding of the structure and function of the TNBC genome, providing a powerful tool for investigating the occurrence and development of diseases. This review focuses on the application of CRISPR/Cas technology in the personalized diagnosis and treatment of TNBC. We begin by discussing the unique attributes of TNBC and the limitations of current diagnostic and treatment approaches: conventional diagnostic methods provide limited insights into TNBC, while traditional chemotherapy drugs are often associated with low efficacy and severe side effects. The CRISPR/Cas system, which activates Cas enzymes through complementary guide RNAs (gRNAs) to selectively degrade specific nucleic acids, has emerged as a robust tool for TNBC research. This technology enables precise gene editing, allowing for a deeper understanding of TNBC heterogeneity by marking and tracking diverse cell clones. Additionally, CRISPR facilitates high-throughput screening to promptly identify genes involved in TNBC growth, metastasis, and drug resistance, thus revealing new therapeutic targets and strategies. In TNBC diagnostics, CRISPR/Cas was applied to develop molecular diagnostic systems based on Cas9, Cas12, and Cas13, each employing distinct detection principles. These systems can sensitively and specifically detect a variety of TNBC biomarkers, including cell-specific DNA/RNA and circulating tumor DNA (ctDNA). In the realm of precision therapy, CRISPR/Cas has been utilized to identify key genes implicated in TNBC progression and treatment resistance. CRISPR-based screening has uncovered potential therapeutic targets, while its gene-editing capabilities have facilitated the development of combination therapies with traditional chemotherapy drugs, enhancing their efficacy. Despite its promise, the clinical translation of CRISPR/Cas technology remains in its early stages. Several clinical trials are underway to assess its safety and efficacy in the treatment of various genetic diseases and cancers. Challenges such as off-target effects, editing efficiency, and delivery methods remain to be addressed. The integration of CRISPR/Cas with other technologies, such as 3D cell culture systems, human induced pluripotent stem cells (hiPSCs), and artificial intelligence (AI), is expected to further advance precision medicine for TNBC. These technological convergences can offer deeper insights into disease mechanisms and facilitate the development of personalized treatment strategies. In conclusion, the CRISPR/Cas system holds immense potential in the precise diagnosis and treatment of TNBC. As the technology progresses and becomes more costs-effective, its clinical relevance will grow, and the translation of CRISPR/Cas system data into clinical applications will pave the way for optimal diagnosis and treatment strategies for TNBC patients. However, technical hurdles and ethical considerations require ongoing research and regulation to ensure safety and efficacy.

Alzheimer's disease (AD) is the commonest neurodegenerative disease that causes memory decline, cognitive dysfunction and behavior disorders in the aged people. Primary pathological hallmarks of AD include amyloid-β (Aβ), neurofibrillary tangles (NFTs), gliosis, and neuronal loss. Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels have important physiological functions, especially in aspects of controlling the resting membrane potential, pacemaker activity, memory formation, sleep and arousal. This article reviews the structure, distribution, regulation of HCN channels and the role of HCN channels in the pathological mechanisms of AD, aiming to provide drug therapeutic targets for the prevention and treatment of AD.
Humans ; Alzheimer Disease/physiopathology* ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels/physiology* ; Animals ; Amyloid beta-Peptides/metabolism*

OBJECTIVE: To explore the integrated traditional Chinese and Western medicine treatment plan for ankylosing spondylitis complicated with lower cervical spine fracture and dislocation, adopt the treatment plan of preoperative continuous traction, intraoperative prizing reduction combined with anterior long-segment plate-screw and posterior short-segment pedicle screw-rod system internal fixation, and evaluate its surgical efficacy and clinical application value. METHODS: From June 2018 to September 2022, 7 male patients with ankylosing spondylitis complicated with lower cervical spine fractures were admitted, aged 43 to 65 years old. Among them, there was 1 case of C_3,4 fracture, 1 case of C_4,5 fracture, 1 case of C_6,7 fracture, and 4 cases of C_5,6 fracture, all of which were fracture and dislocation. All patients received preoperative continuous skull traction, and intraoperative prizing reduction combined with anterior long-segment plate-screw and posterior short-segment pedicle screw-rod system internal fixation. The Neck Disability Index (NDI), Japanese Orthopedic Association (JOA) score, and Frankel scale were used to evaluate the neurological function and quality of life before and after surgery. The visual analogue scale (VAS) was used to evaluate neck and limb pain. The operation time, blood loss, hospital stay, and surgery-related complications were recorded. RESULTS: All 7 patients were followed up for 6 to 24 months after surgery. The operation time of the 7 patients ranged from 300 to 480 minutes, the blood loss ranged from 300 to 1000 ml, and the hospital stay ranged from 8 to 25 days. The preoperative NDI of the 7 patients ranged from 25% to 42%, which decreased to 12% to 30% at 1 week after surgery and 5% to 25% at the last follow-up. The preoperative JOA score ranged from 8 to 13 points, which increased to 12 to 15 points at 1 week after surgery and 13 to 16 points at the last follow-up. The preoperative VAS ranged from 6 to 8 points, which decreased to 2 to 4 points at 1 week after surgery and 0 to 3 points at the last follow-up. Regarding the Frankel grade of neurological function, 2 patients were grade C before surgery and recovered to grade D at the last follow-up after surgery, and the remaining patients recovered to grade E at the last follow-up after surgery. There were 3 cases of pressure ulcers, including 1 case of intraoperative pressure ulcer, 1 case of cervical cerebrospinal fluid leakage, 1 case of screw loosening, and 1 case of aggravated fracture dislocation due to preoperative traction. CONCLUSION Preoperative cervical traction combined with intraoperative prizing reduction and anterior long-segment plate combined with posterior short-segment pedicle screw internal fixation provides a safe and effective surgical option for ankylosing spondylitis complicated with lower cervical spine fracture and dislocation, which can minimize surgical trauma and improve clinical efficacy. However, this study has a small sample size and a short follow-up time for some patients, so further verification with large-sample and long-term follow-up data is still needed.
Humans ; Male ; Adult ; Middle Aged ; Fracture Fixation, Internal/methods* ; Spondylitis, Ankylosing/surgery* ; Cervical Vertebrae/surgery* ; Spinal Fractures/surgery* ; Traction ; Aged ; Joint Dislocations/surgery*

OBJECTIVE: To investigate the clinical efficacy and safety of single Kirschner wire assisted poking and closed reduction in the treatment of Gartland type Ⅲ supracondylar humeral fractures in children. METHODS: A retrospective analysis was performed on patients diagnosed with Gravland type Ⅲ supracondylar humeral fractures between January 2022 and June 2023. A total of 46 patients were treated with closed reduction assisted by Kirschner wires and percutaneous Kirschner wire internal fixation.There were 25 males and 21 females. The age ranged from 5 to 10 years old, with an average of (5.8±1.8) years old. The left side was involved in 28 patients and the right side in 18 patients. Record the operative duration for patients, the number of fluoroscopic exposures, fracture healing time, postoperative carrying angle, Baumann angle, elbow joint function score at three months post-operation, and any associated complications. RESULTS: All 46 patients were followed up for a period of 12 to 16 weeks, with an average of (13.74±1.44 )weeks. The operation duration was (30.7±5.1) minutes, the fluoroscopy count was (10.2±2.7) times, the postoperative carrying angle of the elbow joint was (8.7±2.2) degrees, and the Baumann angle was (71.5±2.9) degrees. All fractures achieved successful union in all patients, with a mean healing time of (25.5±1.7) days.At the final follow-up, elbow joint function was assessed using the Flynn criteria, with 43 patients rated as excellent and 3 patients rated as good. No complications were observed, including cubitus varus, nerve injury, or local infection. CONCLUSION The use of a single Kirschner wire assisted prying reduction for treating Gartland type Ⅲ supracondylar humeral fractures in children demonstrates excellent clinical efficacy and safety.
Humans ; Male ; Female ; Child ; Bone Wires ; Child, Preschool ; Humeral Fractures/physiopathology* ; Retrospective Studies ; Fracture Fixation, Internal/instrumentation* ; Treatment Outcome ; Fracture Healing

Objective To investigate the clinical efficacy of continuous irrigating mode of endoscopic ear sur-gery(CIM-EES)utilizing tragus cartilage-perichondrium without tympanomeatal flap elevation in repairing tympanic membrane perforation.Methods The date of 70 patients(70 ears)who underwent tympanic membrane repair under ear endoscopy from June 2020 to August 2023 were randomly selected and analyzed.They were divided into two groups according to the time of operation:CIM-EES group(observation group)of 44 cases(44 ears)and a conven-tional surgery group(control group)of 26 cases(26 ears).Both groups were repaired with tympanic membrane per-foration using the method of implanting tragus cartilage-perichondrium grafts under endoscope without tympa-nomeatal flap elevation.After 6 months of postoperative follow-up,two groups were compared in terms of operation time,frequency of the endoscopic lens scrubbing,postoperative healing rate,and hearing improvement outcomes.Results The average operation time of the observation group and the control group was 37.50±4.81 minutes and 50.31±8.21 minutes respectively,and the average number of scrubbing the endoscope was 6.77±1.51 and 35.54±7.13 respectively,there was statistical difference significance between the two groups(P＜0.01).All patients in the observation group were successfully repaired in the first stage,with the healing rate of 100％(44/44),and one patient in the control group had a postoperative small perforation that healed after secondary repair with the healing rate of 96.15％(25/26).There was no statistically significant difference between the two groups.The average 6 months postoperative air conduction threshold and air-bone conduction threshold of 0.5-4 kHz in the two groups improved compared to preoperative results(P＜0.01),with no statistically significant difference between the two groups(P＞0.05).Conclusion Endoscopic myringoplasty without tympanomeatal flap elevation under continuous irrigating mode has the advantages of high healing rate,short operation time,simple surgical operation,few post operation complications and good hearing improvement.

Objective:To establish a model for the combined detection of serum copeptin and inflammatory markers in acute stroke (AS), and to explore the value of copeptin and inflammatory marker detection in the clinical diagnosis and prognosis assessment of AS.Methods:A total of 75 patients were diagnosed with acute ischemic stroke (AIS) [46 males, age (64.1±11.7) years] and 45 patients with acute intracerebral hemorrhage (ICH) [28 males, age (61.0±13.9) years] who were admitted to the First Affiliated Hospital of Hunan University of Chinese Medicine through the emergency department from January 1 to July 31, 2024, were included as the observation group. Meanwhile, 60 healthy individuals [39 males, age (64.4±8.2) years] were selected as the control group (HC). The differences in serum copeptin levels and inflammatory markers among different groups were compared. ROC curves were drawn to analyze the value of copeptin and inflammatory markers in the clinical diagnosis and prognosis assessment of AIS. The Kaplan-Meier method was used to draw survival curves to analyze the in-hospital survival rates of patients in different groups. Cox regression analysis was conducted to identify the risk factors affecting the prognosis of AIS patients.Results:The level of copeptin was significantly elevated in AS, with the results showing ICH>AIS>HC ( H=100.11, P<0.001). Copeptin demonstrated the highest efficacy in the early diagnosis of AIS and ICH (AUC=0.893, sensitivity 89.3%, specificity 75.0%; AUC=0.986, sensitivity 95.6%, specificity 93.3%) and the assessment of prognosis (AUC=0.997, sensitivity 100%, specificity 96.8%; AUC=0.907, sensitivity 86.7%, specificity 86.7%), outperforming other single indicators. The combined detection of copeptin with the neutrophil-to-lymphocyte ratio (NLR) and the systemic immune-inflammation index (SIIRI) was the best combination for the early diagnosis of AIS and ICH (AUC=0.937, sensitivity 77.3%, specificity 98.3%; AUC=0.989, sensitivity 95.6%, specificity 95.0%) and for the assessment of prognosis (AUC=0.996, sensitivity 100%, specificity 96.8%; AUC=0.944, sensitivity 86.7%, specificity 90.0%). Kaplan-Meier survival curves showed that AIS patients in the low-value group of copeptin and NLR had a higher survival rate during hospitalization than those in the high-value group ( HR 54.46, 7.608, P<0.01, respectively), and ICH patients in the low-value group of copeptin, SIIRI, SIRI, and SII had a higher survival rate during hospitalization than those in the high-value group ( HR 12.67, 7.923, 3.567, 5.925, P<0.05); Cox regression showed that copeptin, NLR, NIHSS, and mRS were independent risk factors affecting the prognosis of patients with AIS ( HR 1.421, 1.368, 1.158, and 1.188, respectively, P<0.05), copeptin and SIIRI were independent risk factors affecting the prognosis of ICH ( HR 1.308, 1.113, P<0.05), and GCS was a protective factor affecting ICH prognosis ( HR=0.741, P<0.05). Conclusion:Copeptin and inflammatory indicators can reflect the severity of different subtypes of stroke. The single or combined detection shows good value in the clinical application of AS. The combination of copeptin-NLR and copeptin-SIIRI is respectively the best comprehensive biomarker combination for the early diagnosis and prognosis assessment of AIS and ICH.

Objective:This study aimed to evaluate to evaluate the performance of equilibrium dialysis combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the accurate measurement of free testosterone in clinical samples. Mthods We conducted a prospective observational study using 161 serum samples from healthy women of reproductive 26(24, 32)years at the Gynecology Outpatient department of Peking Union Medical College Hospital from June to September 2024, and their concentrations were determined. In this study, after equilibrium dialysis of serum samples, free testosterone was extracted from the dialysate using a magnetic bead-based method. It was then directly derivatized using hydroxylamine hydrochloride in situ after elution from the magnetic bead and further quantified by LC-MS/MS.Method:validation assessed linearity, limit of quantification (LOQ), precision, accuracy, matrix effects, and carryover according to established guidelines. Data were analyzed using Origin 2019 and WPS Office 2019.Results:The method demonstrated excellent linearity ( R2>0.99) across 1-250 pg/ml with an LOQ of 1 pg/ml. The coefficients of variation for both intra-day and inter-day imprecision were less than 10% while recovery rates ranged from 92.60% to 99.10%. Matrix effect deviations were all within the range of 6% and carryover was negligible. Conclusions:In this study, the established method of magnetic bead-based extraction followed by in situ derivatization combined with liquid chromatography-tandem mass spectrometry performed well, and could be further applied to the detection of free testosterone concentration in childbearing age women.

AIM To study the chemical constituents from the buds of Aralia chinensis L.var.nuda Nakai and their in vitro anti-inflammatory activities.METHODS The 70％ethanol extract from the buds of A.chinensis var.nuda was isolated and purified by silica gel,Sephadex LH-20,ODS and semi-preparative HPLC,then the structures of compounds were identified by physicochemical properties and spectral data.Their anti-inflammatory activities in vitro were evaluated by RAW264.7 model.RESULTS Sixteen compounds were isolated and identified as 4-(2,2-dibutoxyethyl)phenol(1),trans-linalool-3,7-oxide-6-O-β-D-glucopyranoside(2),2'-O-(9Z,12Z,15Z-octadecatrienoyl)glyceryl β-D-galactopyranoside(3),quercetin-3-O-β-D-glucopyranoside(3'→ O-3''')quercetin-3-O-β-D-galactopyranoside(4),syringaresinol-4'-O-β-D-glucopyranoside(5),p-hydroxybenzaldehyde(6),7α-hydroxystigmasterol 3-O-β-D-glucopyranoside(7),trans-p-hydroxy cinnamic acid methyl ester(8),funingensin A(9),3,4-dihydroxy-acetophenone(10),N-acetyltyramine(11),3,4-di-O-caffeoyl quinic acid(12),chlorogenic acid(13),aralia cerebroside(14),caffeic acid methyl ester(15),tetradecanoic acid(16).The IC50values of compounds 8,10,12 and 13 were(22.19±1.59),(35.25±1.30),(13.38±0.72),(15.73±1.16)μmol/L,respectively.CONCLUSION Compound 1 is a new compound,2-13 are isolated from genus Aralia for the first time.Compounds 8,10,12,13 exhibit significant in vitro anti-inflammatory activities.

PURPOSE: Septic shock is associated with high mortality and poor outcomes among sepsis patients with coagulopathy. Although traditional statistical methods or machine learning (ML) algorithms have been proposed to predict septic shock, these potential approaches have never been systematically compared. The present work aimed to develop and compare models to predict septic shock among patients with sepsis. METHODS: It is a retrospective cohort study based on 484 patients with sepsis who were admitted to our intensive care units between May 2018 and November 2022. Patients from the 908th Hospital of Chinese PLA Logistical Support Force and Nanchang Hongdu Hospital of Traditional Chinese Medicine were respectively allocated to training (n=311) and validation (n=173) sets. All clinical and laboratory data of sepsis patients characterized by comprehensive coagulation indexes were collected. We developed 5 models based on ML algorithms and 1 model based on a traditional statistical method to predict septic shock in the training cohort. The performance of all models was assessed using the area under the receiver operating characteristic curve and calibration plots. Decision curve analysis was used to evaluate the net benefit of the models. The validation set was applied to verify the predictive accuracy of the models. This study also used Shapley additive explanations method to assess variable importance and explain the prediction made by a ML algorithm. RESULTS: Among all patients, 37.2% experienced septic shock. The characteristic curves of the 6 models ranged from 0.833 to 0.962 and 0.630 to 0.744 in the training and validation sets, respectively. The model with the best prediction performance was based on the support vector machine (SVM) algorithm, which was constructed by age, tissue plasminogen activator-inhibitor complex, prothrombin time, international normalized ratio, white blood cells, and platelet counts. The SVM model showed good calibration and discrimination and a greater net benefit in decision curve analysis. CONCLUSION The SVM algorithm may be superior to other ML and traditional statistical algorithms for predicting septic shock. Physicians can better understand the reliability of the predictive model by Shapley additive explanations value analysis.
Humans ; Shock, Septic/blood* ; Machine Learning ; Male ; Female ; Retrospective Studies ; Middle Aged ; Aged ; Sepsis/complications* ; ROC Curve ; Cohort Studies ; Adult ; Intensive Care Units ; Algorithms ; Blood Coagulation ; Critical Illness

OBJECTIVE: To explore the effect of acupuncture therapy on dysphagia in patients with Parkinson's disease. METHODS: This randomized controlled study lasted 42 days and included 112 patients with Parkinson's disease and dysphagia. Participants were randomly assigned to the experimental and control groups (56 cases each group) using the completely randomized design, all under routine treatment. The experimental group was given acupuncture therapy. The primary outcome was Penetration-Aspiration Scale (PAS). The secondary outcomes were (1) Standardized Swallowing Assessment (SSA), and (2) nutritional status including body mass index (BMI), serum albumin, prealbumin, and hemoglobin. Adverse events were recorded as safety indicators. RESULTS: One participant quitted the study midway. There were no significant differences in baseline assessment (P>0.05). After treatment, both groups showed significant improvement in PAS, SSA and nutritional status except for BMI of the control group. There were significant differences between the two groups in the PAS for both paste and liquid, SSA (25.18±8.25 vs. 20.84±6.92), BMI (19.97±3.34 kg/m²vs. 21.26 ±2.38 kg/m²), serum albumin (35.16 ±5.29 g/L vs. 37.24 ±3.98 g/L), prealbumin (248.33 ±27.72 mg/L vs. 261.39 ±22.10 mg/L), hemoglobin (119.09±12.53 g/L vs. 126.67±13.97 g/L) (P<0.05). There were no severe adverse events during the study. CONCLUSION: The combination of routine treatment and acupuncture therapy can better improve dysphagia and nutritional status in patients with Parkinson's disease, than routine treatment solely. (registration No. CLINICALTRIAL gov NCT06199323).
Humans ; Parkinson Disease/therapy* ; Deglutition Disorders/physiopathology* ; Acupuncture Therapy/adverse effects* ; Male ; Female ; Aged ; Middle Aged ; Treatment Outcome ; Nutritional Status ; Body Mass Index