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.

ObjectiveTo collate and compare the characteristics and differences in the methods for constructing animal models of different subtypes of gastroesophageal reflux disease （GERD） based on literature， providing a reference for researchers in this field regarding animal model construction. MethodsExperimental studies related to GERD including reflux esophagitis （RE）， nonerosive reflux disease （NERD） and Barrett's esophagus （BE） model construction from January 1， 2014 to January 27， 2024， were retrieved from databases such as CNKI， Wanfang， VIP， Web of Science， and Pubmed. Information on animal strains， genders， modeling methods including disease-syndrome combination models， modeling cycles were extracted； for studies with model evaluation， the methods of model evaluation were also extracted； then analyzing all those information. ResultsA total of 182 articles were included. SD rats were most frequently selected when inducing animal models of RE （88/148， 59.46%） and NERD （9/14， 64.29%）. For BE， C57BL/6 mice were most commonly used （11/20， 55.00%）. Male animals （RE： 111/135， 82.22%； NERD： 11/14， 78.57%； BE： 10/12， 83.33%） were the most common gender among the three subtypes. The key to constructing RE animal models lies in structural damage to the esophageal mucosal layer， gastric content reflux， or mixed reflux， among which forestomach ligation + incomplete pylorus ligation （42/158， 26.58%） was the most common modeling method； the key to constructing NERD animal models lies in micro-inflammation of the esophageal mucosa， visceral hypersensitivity， and emotional problems， and intraperitoneal injection of a mixed suspension of ovalbumin and aluminum hydroxide combined with acid perfusion in the lower esophagus （8/14， 57.14%） was the most common modeling method； the key to constructing BE animal models lies in long-term inflammatory stimulation of the esophageal mucosa and bile acid reflux， and constructing interleukin 2-interleukin 1β transgenic mice （7/25， 28.00%） was the most common modeling method. Adverse psychological stress was the most common method for inducing liver depression. ConclusionsThe construction key principles and methodologies for RE， NERD， and BE animal models exhibit significant differences. Researchers should select appropriate models based on subtype characteristics （e.g.， RE focusing on structural damage， NERD emphasizing visceral hypersensitivity）. Current studies show insufficient exploration of traditional Chinese medicine disease-syndrome combination models. Future research needs to optimize syndrome modeling approaches （e.g.， composite etiology simulation） and establish integrated Chinese-Western medicine evaluation systems to better support mechanistic investigations of traditional Chinese medicine.

Objective:To evaluate the effect of five-element acupuncture on clinical symptoms, brain metabolism and immunoglobulin level in patients with chronic fatigue syndrome.Methods:It was a randomized controlled trial. A total of 94 patients with chronic fatigue syndrome in our hospital from January 2021 to January 2022 were selected and divided into two groups according to the random number table method, with 47 in each group. The conventional western medicine group was treated with conventional western medicine, and the five-element acupuncture group was treated with five-element acupuncture on the basis of the conventional western medicine group. Both groups were treated for 4 weeks. Before and after treatment, the serum levels of interferon-γ (IFN-γ), corticosterone (CORT), IL-2 and 5-hydroxytryptamine (5-HT) were detected by ELISA; the levels of natural killer (NK) cells, CD4 +, CD8 +, IgG and IgM were detected by flow cytometry; the whole body superconducting MRI scanner was used to scan T2 Flair, T2WI and TlWI sequences of the hippocampus, and the spectral curves and the areas under the peak of N-acetylaspartic acid (NAA), creatine (Cr) and choline (Cho) were obtained, and the ratios of Cho/Cr and NAA/Cr were calculated. the fatigue Scale-14 (FS-14) and Fatigue Severity Scale (FSS) were used to evaluate the fatigue state of the patients, and the Depression-Anxiety-Stress Scale-21 (DASS-21) and Beck Anxiety Inventory (BAI) were used to evaluate the anxiety state of the patients. Symptom Checklist 90 (SCL-90) and Somatic and Mental Health Report Score (SPHERE) were used to evaluate the quality of life of patients. And the clinical efficacy was evaluated. Results:After treatment, the levels of IgG, CD4 + and NK in the five-element acupuncture group were significantly higher than those in the conventional western medicine group ( t values were 4.76, 3.65, 6.42, respectively, P<0.01), and the level of IgM, CD8 + was significantly lower than that in the conventional western medicine group ( t values were 7.30, 4.79, P<0.01); the levels of IFN-γ, IL-2 and 5-HT in the observation group were significantly higher than those in the conventional western medicine group ( t values were 7.60, 4.05, 2.79, respectively, respectively, P<0.01), and the level of CORT was significantly lower than that in the conventional western medicine group ( t=6.72, P<0.01); the NAA/Cr levels in the left [(1.10±0.04) vs. (1.05±0.03), t=6.86] and right [(1.18±0.02) vs. (1.21±0.03), t=8.23] hippocampus of the experimental group were significantly higher than those in the conventional western medicine group ( P<0.01), and the Cho/Cr levels in the left [(1.08±0.04) vs. (1.03±0.03), t=5.70] and right [(1.17±0.02) vs. (1.20±0.03), t=5.71] hippocampus of the experimental group were significantly lower than those of the conventional western medicine group ( P<0.01). After treatment, the scores of physical fatigue, mental fatigue and FSS in the five-element acupuncture group were significantly lower than those in the conventional western medicine group ( t values were 8.08, 9.08 and 7.07, respectively, P<0.01). The scores of DASS-21, BAI, SCL-90 and SPHERE in the conventional western medicine group were significantly lower than those in the conventional western medicine group ( t values were 3.63, 5.77, 8.74, 5.92, respectively, P<0.01).The total effective rate was 95.74% (45/47) in the five-element acupuncture group and 82.98% (39/47) in the conventional western medicine group, and there was no significant difference between the two groups ( χ2=2.80, P=0.094). Conclusion:Five-elements acupuncture can improve the expression of T lymphocytes, increase the levels of immunoglobulin and NK, reduce the level of CORT, regulate the brain metabolism of NAA in the left and right hippocampus, improve the clinical symptoms and negative emotions, and improve the clinical efficacy and quality of life in patients with chronic fatigue syndrome.

Objective To design an intelligent surgery management system for closed-loop management of the entire surgical process.Methods An intelligent surgery management system was developed with digital twin technology,Microsoft Visual Studio 12.0 development toolset and C# language,which realized multi-modal data acquisition with IoT devices such as radio frequency identification(RFID)bracelets,mobile terminals and positioning base stations and real-time data mirroring and gathering by change data capture(CDC)technology.There were four functional modules included in the system developed,including the modules for IoT device management,transfer process recording,intelligent management command chamber and surgical progress reminder.Results The system developed could provide real-time feedback on the operation status of all aspects of surgery,shorten the lengths of preoperative positioning and surgical reception and improve the satisfaction of both doctors and patients.Conclusion The system developed achieves closed-loop management and complete data traceability of the entire surgical process,helping to form an efficient and safe surgical operation management system.[Chinese Medical Equipment Journal,2024,45(1):42-46]

Cellulose synthase (CesA), one of the key enzymes in the biosynthesis of cellulose in plants, plays an important role in plant growth and plant resistance. In this study, a total of 21 AsCesA genes from Aquilaria sinensis were systematically identified and the physico-chemical characteristics were analyzed based on genome database and bioinformatical methods. The phylogenetic tree was constructed and the gene location on chromosome, cis-acting elements in the 2 000 basepairs upstream regulatory regions and conservative motifs were analyzed. The AsCesA proteins were mainly located on the plasma membrane. The number of amino acids of the proteins ranged from 390 to 1 261. The isoelectric point distributed from 5.67 to 8.86. All of the 21 AsCesA proteins possessed the transmembrane domains, the number of which was from 6 to 8. The genes were classified into 3 groups according to the phylogenetic relationship. Obvious differences were observed in motif composition in genes from different groups. However, motif2, motif6, motif7 and motif10 were observed in all of AsCesA proteins. Analysis of cis-acting elements indicated that AsCesA genes family has cis-acting elements related to plant hormones, abiotic stresses, and biological processes. Seven AsCesA genes with differential expression were selected according to the calli transcriptome data induced by NaCl at different times and their expression levels under different abiotic stresses were analyzed by quantitative real-time PCR. The results indicated that salt, low temperature, drought, and heavy metal stresses could affect the expression level of AsCesA genes, and the abundance of AsCesA1, AsCesA3 and AsCesA20 showed a significant change, implying their potential important roles to the abiotic stresses. The accumulation pattern of cellulose content under different abiotic stresses was similar to the expression trend of AsCesA genes. Our results provide valuable insights into the role of cellulose synthase in A.sinensis in plant defense.

Background::Although overnight fasting is recommended prior to endoscopic retrograde cholangiopancreatography (ERCP), the benefits and safety of high-carbohydrate fluid diet (CFD) intake 2 h before ERCP remain unclear. This study aimed to analyze whether high-CFD intake 2 h before ERCP can be safe and accelerate patients’ recovery.Methods::This prospective, multicenter, randomized controlled trial involved 15 tertiary ERCP centers. A total of 1330 patients were randomized into CFD group ( n = 665) and fasting group ( n = 665). The CFD group received 400 mL of maltodextrin orally 2 h before ERCP, while the control group abstained from food/water overnight (>6 h) before ERCP. All ERCP procedures were performed using deep sedation with intravenous propofol. The investigators were blinded but not the patients. The primary outcomes included postoperative fatigue and abdominal pain score, and the secondary outcomes included complications and changes in metabolic indicators. The outcomes were analyzed according to a modified intention-to-treat principle. Results::The post-ERCP fatigue scores were significantly lower at 4 h (4.1 ± 2.6 vs. 4.8 ± 2.8, t = 4.23, P <0.001) and 20 h (2.4 ± 2.1 vs. 3.4 ± 2.4, t= 7.94, P <0.001) in the CFD group, with least-squares mean differences of 0.48 (95% confidence interval [CI]: 0.26–0.71, P <0.001) and 0.76 (95% CI: 0.57–0.95, P <0.001), respectively. The 4-h pain scores (2.1 ± 1.7 vs. 2.2 ± 1.7, t = 2.60, P = 0.009, with a least-squares mean difference of 0.21 [95% CI: 0.05–0.37]) and positive urine ketone levels (7.7% [39/509] vs. 15.4% [82/533], χ2 = 15.13, P <0.001) were lower in the CFD group. The CFD group had significantly less cholangitis (2.1% [13/634] vs. 4.0% [26/658], χ2 = 3.99, P = 0.046) but not pancreatitis (5.5% [35/634] vs. 6.5% [43/658], χ2 = 0.59, P = 0.444). Subgroup analysis revealed that CFD reduced the incidence of complications in patients with native papilla (odds ratio [OR]: 0.61, 95% CI: 0.39–0.95, P = 0.028) in the multivariable models. Conclusion::Ingesting 400 mL of CFD 2 h before ERCP is safe, with a reduction in post-ERCP fatigue, abdominal pain, and cholangitis during recovery.Trail Registration::ClinicalTrials.gov, No. NCT03075280.

Prosthodontic treatment is being performed for morphology and functional restoration due to damage and loss of teeth. As the aesthetic demands of patients increase, interest in ceramic materials with shades and translucency similar to natural teeth has increased.Recently, the manufacturing and processing technology of ceramic materials has greatly improved, and the market for dental ceramic materials is growing rapidly. The purpose of this literature review and evaluation is to provide information on the classification and properties of dental ceramic materials with excellent aesthetics and fracture resistance. In this article, it is classified as follows: I) Dental porcelain; II) Sinterable all-ceramic; III) Glass-ceramic for casting; IV) Glass-infiltrated alumina ceramic; V) Glass-ceramic ingots for heat-pressing technique; Vl) Blocks for CAD/CAM; Vll) Ceramic for CAD/3D printing. Dental ceramic materials and their restoration manufacturing methods have evolved significantly over the past decade. As a result, the manufacturing method of restorations has progressed from the layered firing technique of powdered materials or heat-pressing technique to the cutting and processing of single and multi-layer blocks using CAD/CAM technology, leading to the introduction of CAD/3D printing technology. In this manuscript, we will review the types of ceramic materials used in the fabrication of dental restorations and their advantages and disadvantages.

Brasilicardin A, a diterpene glycoside isolated from pathogenic actinomycete Nocardia brasiliensis IFM 0406, has become a novel immunosuppressant candidate due to its significant immunosuppressive activity, low toxicity and unique mechanism of action. However, brasilicardin A and its analogues have become a research hotspot to the development of this promising immunosuppressant because of the low-yield production in the natural pathogenic producer and the synthetically challenging skeleton. According to the reported biosynthetic pathway of brasilicardin A, the function of involved diterpene synthase was analyzed by bioinformatics. Then the genes bra1-5 that synthesize the brasilicardin A skeleton were directionally amplified from the pathogenic strain N. brasiliensis IFM 0406, and heterologous expression was achieved successfully in Streptomyces albus R1. The compounds were isolated and purified by using various column chromatographies including silica gel column chromatography and semi-preparative HPLC. Six new brasilicardins were established and named brasilicardin H-M. The activity of brasilicardins was screened using lipopolysaccharide (LPS)-activated mouse primary macrophage inflammation model. Brasilicardin H-M exhibited good inhibitory activity on nitric oxide (NO) release with IC₅₀ values of 28.24 ± 3.70, 37.44 ± 2.00, 39.85 ± 4.02, 26.77 ± 4.40, 65.25 ± 1.48 and 15.24 ± 2.72 μmol·L^-1, respectively (indomethacin as the positive control with IC₅₀ value of 34.28 ± 4.10 μmol·L^-1). The results indicated that six compounds had potential anti-inflammatory activity. This study laid a foundation for the elucidation of the brasilicardin A biosynthetic pathway and evaluation of the structure-activity relationship as well as new drug developments.

Objective To compare the effects of three kinds of artificial hydronephrosis in percutaneous nephrolithotomy.Methods A total of 120 patients who underwent single-tract percutaneous nephrolithotomy in the Eighth Hospital of Wuhan from May 2020 to April 2022 were selected and divided equally into three groups according to different methods of artificial hydronephrosis.Patients in group A were received preoperative indwelling ureteral catheter and injection of normal saline through ureteral catheter to dilate renal pelvis and form artificial hydronephrosis.Patients in Group B were placed with double J catheters before surgery,and the bladder was filled with physiological saline through the catheter.The renal pelvis was dilated through the double J catheters,resulting in artificial hydronephrosis.Patients in Group C were received intravenous injection of furosemide and stimulated diuretic method to actively dilate renal pelvis to form hydronephrosis.The one-time puncture success rate,channel establishment time,overall operation time,stone clearance rate and incidence of surgical complications of percutaneous nephrolithotomy after hydronephrosis were compared among the three groups.Results The operation was successfully completed in the three groups.There was no significant difference in the one-time puncture success rate and channel establishment time between group A and group B(P>0.05),which were all higher than group C(P<0.05).The overall operation time of group B was shorter than that of group A and group C(P<0.05).There were no significant differences in stone clearance rate and surgical complications among the three groups(P>0.05).Conclusion Preoperative indwelling of double J tubes to create artificial kidney hydronephrosis has advantages such as high success rate of one-time puncture,short channel establishment time,and surgical time.

To develop a rapid differential detection method for porcine epidemic diarrhea virus(PEDV)and transmissible gastroenteritis virus(PEDV),M gene sequences of PEDV and TGEV were compared,the inner and outer primer pairs and probes were designed according to the highly conserved region.PEDV-Probe was labeled with FAM at5'end and BHQ1 at 3'end.TGEV-Probe was labeled with CY5.5 at the 5'end and BHQ2 at the 3'end.After optimizing the reaction condi-tions and system,a dual fluorescent RT-LAMP assay for PEDV and TGEV rapid identification was established.The amplification could be completed within 60 min in a 63 ℃ water bath and then stopped at 85 ℃ for 10 min.Then the tubes were placed in a multicolor imaging system,and the re-sults could be observed under 520 nm and 690 nm dual channels.There was no cross-reaction when other common swine viral pathogens were detected by this method.The sensitivity of the assay was evaluated with a 10-fold diluted recombinant plasmid as templates.The lowest detection limit was 102 copies/μL recombinant plasmid,which was 10 times more sensitive than the conventional RT-PCR method.Seventy-two PEDV-positive samples,49 TGEV-positive samples,and 40 PEDV and TGEV co-infected samples were detected from 175 anal swab samples of diarrheic piglets by the established method,which were all higher than the detection rates of the conventional RT-PCR method.The dual fluorescent RT-LAMP method established in this study can be used to amplify the target gene in an ordinary water bath without gel electrophoresis,which provides technical sup-port for rapid and convenient differential diagnosis of PED and TGE and simultaneous detection of PEDV and TGEV co-infection.