Traditional Chinese medicine （TCM） diagnostics is a discipline that studies the basic theories and fundamental skills of diagnostic methods， disease diagnosis， and differentiation in accordance with the theories of TCM. The artificial intelligence （AI） technology has gained remarkable achievements in the intelligentization of the four diagnostic methods in TCM and the standardization of differentiation and diagnosis. However， it still faces many challenges. The standardization of clinical data collection is difficult， and the data quality is uneven， which affects the usability of the data. The integration of the four diagnostic information is insufficient. Most instruments can only collect data from a single diagnostic method， lacking overall integrity. The scientific nature of the diagnostic model needs to be improved. The existing models lack dynamics and the reasoning logic of TCM differentiation. The accuracy of intelligent methods needs to be improved， and the existing evaluation indicators cannot fully reflect the practical application effect of the model. Furthermore， the relevant laws and regulations are still not perfect， and data security and patient privacy lack guarantees. The cultivation of compound talents is insufficient， and there is a lack of interdisciplinary talents who are proficient in both TCM and AI. On this basis， this paper expounded on the current development status， difficulties， and bottlenecks of AI in TCM diagnosis and then explored the development trend of AI in the field of TCM diagnosis. It proposed solutions such as optimizing the data collection process， constructing multimodal diagnostic models， facilitating multi-disciplinary exchanges and cooperation， improving laws and regulations， and cultivating compound talents. It is hoped that modern， standardized， normalized， and intelligent TCM diagnosis can be further promoted， thereby providing new impetus and methods for the inheritance and innovation of TCM.

Objective: To develop a RHCE genotyping assay based on kompetitive allele-specific PCR (KASP) and assess its clinical accuracy for RhCE blood group determination. Methods: KASP primers were designed to interrogate three RHCE loci: the 109 bp insertion/deletion in intron 2, c. 307T>C, and c. 676C>G. A total of 1 194 RhD-positive inpatients from Chengdu were typed by both KASP genotyping and manual tube serology. Discordant samples (n=10) were retested by both methods and further resolved by Sanger sequencing. An additional 377 cases were tested for the c. 48C>G locus to evaluate the predictive accuracy of individual loci and combined locus testing for RhC antigen. Results: Genotyping concordance with serology was 100.0% for both the c. 676C>G locus (RhE/Rhe) and the c. 307T>C locus (Rhc). For RhC prediction using the 109 bp insertion, overall accuracy was 99.7% (1 191/1 194); the 3 discordant cases were confirmed by Sanger sequencing to be false negatives attributable to 109 bp deletion in intron 2. Testing the c. 48C>G allele for RhC prediction yielded 7 false positives, with an accuracy of 98.1% (370/377). RhC antigen status was determined by combining the 109 bp insertion and the c. 48C allele. After excluding 10 samples with inconsistent results between the two loci, the accuracy reached 100% in the remaining 367 samples. When both loci were applied in combination, accuracy reached 100% in the 367 cases with concordant results. Among the 1 194 patients, CCee (45.8%) and CcEe (31.7%) were the most common RhCE phenotypes. The e antigen had the highest positivity rate (92.2%), and the Ce haplotype was the most frequent (66.9%). Conclusion: The KASP-based RHCE genotyping method achieves high accuracy for clinical RhCE typing. Combining the 109 bp insertion/deletion with the c. 48C allele significantly improves RhC antigen prediction compared with either locus alone. This method was applied to RhCE genotyping of 1 194 RhD-positive inpatients in Chengdu, providing local RhCE phenotype and haplotype distribution data to support RhCE-matched transfusion practice.

Turning to critical illness is a common stage of various diseases and injuries before death. Patients usually have complex health conditions, while the treatment process involves a wide range of content, along with high requirements for doctor′s professionalism and multi-specialty teamwork, as well as a great demand for time-sensitive treatments. However, this is not matched with critical care professionals and the current state of medical care in China. Telemedicine, which shortens the distance of medical professionals and the gap of disease diagnosis and treatments in various regions through electronic information, can effectively solve the current problem. Therefore, there is an urgent need to develop a standardized, high-quality visualization telemedicine round system .Therefore, experts have been organized to search domestic and foreign literature on telemedicine round for critically ill patients and to form this consensus based on clinical experiences so as to further improve the level of critical care treatments in regions.

Objective:To evaluate the lung ultrasound characteristics of mycoplasma pneumoniae pneumonia in children and to investigate the value of lung ultrasound monitoring in clinical diagnosis and treatment.Methods:A retrospective analysis of 62 children with mycoplasma pneumoniae pneumonia admitted to Xi'an Chest Hospital from 7 November to 30 November 2023 was performed，and the characteristic parameters of bedside lung ultrasound and their related clinical data were collected. Pathological lung ultrasound features such as interrupted pleural line，well-spaced B-lines，coalescent B-lines，small subpleural patchy pulmonary consolidation，large pulmonary consolidation and pleural effusion in 12 scan areas of both lungs were observed. The maximum upper and lower diameters，right and left diameters，and anterior and posterior diameters of the large pulmonary consolidations were measured，and the changes in the above signs before and after treatment were measured and compared.Results:In sixty-two children with mycoplasma pneumoniae pneumonia，including 32 males and 30 females，with a mean age of（8.18 ± 2.05）years old and a mean hospital stay of（8.79 ± 2.93）days，lung ultrasound showed interrupted pleural line，well-spaced B-lines，coalescent B-lines，small subpleural patchy pulmonary consolidation，large pulmonary consolidation and pleural effusion，with the incidence of 93.5%（58 /62），33.9%（21/62），32.3%（20/62），59.7%（37/62），66.1%（41/62）and 17.7%（11/62），respectively，in which the large pulmonary consolidations presented rich blood supply were more common in the L6 and L4 areas，while the pleural effusions were more common in the L6 area.The signs of interrupted pleural line，coalescent B-lines，large pulmonary consolidation and pleural effusion were significantly improved after treatment compared with before treatment（all P<0.05）. The upper and lower diameters，left and right diameters，and anterior and posterior diameters of large pulmonary consolidations were significantly reduced after treatment compared with before treatment［（4.19 ± 2.42）cm vs.（2.84 ± 2.31）cm， t=2.613， P=0.011；（2.80 ± 1.82）cm vs.（1.96 ± 1.62）cm， t=2.226， P=0.029；（3.41 ± 2.11）cm vs.（2.12 ± 1.82）cm， t=2.972， P=0.004］.With the process of treatment，the dynamic observation of lung ultrasound showed that the well-spaced B-lines/coalescent B-lines gradually decreased until they completely disappeared or a small number of B-lines remained，and the area of the large pulmonary consolidation showed a dynamic downward trend（all P<0.001），and the area of large pulmonary consolidations gradually decreased until they completely disappeared or only small subpleural patchy pulmonary consolidations and well-spaced/coalescent B-lines remained，and at the same time，the pleural effusion gradually absorbed until it disappeared. Conclusions:Lung ultrasound can detect the distribution area of lung lesions，morphology and blood supply characteristics of children with mycoplasma pneumoniae pneumonia，as well as the dynamic changes after treatment，and lung ultrasound can dynamically monitor and evaluate the progression and regression of the disease in real time，providing a reliable imaging evidence for clinical practice.

AIM To study the chemical constituents from ethyl acetate fraction of Balanophora harlandii Hook.f.and their tyrosinase inhibitory activity.METHODS Separation and purification were performed using silica gel,MCI,ODS,Sephadex LH-20 and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The monophenolase inhibitory activity was determined by the tyrosinase-catalyzed oxidation of L-tyrosine.RESULTS Twenty-four compounds were isolated and identified as sesamin(1),methyl caffeate(2),quercetin(3),5,7-dihydroxychromanone(4),methyl 3,4-dihydroxybenzoate(5),esculetin(6),kaempferol(7),naringenin(8),pyrogallic acid(9),pinosylvin(10),methyl propionate(11),caffeic acid(12),saccharinol(13),ferulic acid(14),trans-p-hydroxycinnamic acid(15),cinnamic acid(16),vanillic acid(17),vanillin(18),4-hydroxyacetophenone(19),4-hydroxybenzaldehyde(20),apigenin(21),(-)-isolariciresinol(22),(-)-secoisolariciresinol(23)and meso-2,3-di(3′,4′-methylenedioxybenzyl)butane-1,4-diol(24).The IC50 values of compounds 3,5,7,8,19,and 20 ranged from(0.246 5±0.028 3)to(1.278 2±0.021 3)mmol/L.CONCLUSION Compounds 1-9、11、15、17-21、24 are isolated from this plant for the first time,and 1,6,9,17-19,24 are first isolated from genus Balanophora.Compounds 3、5、7、8、19 and 20 have tyrosinase inhibitory activity.

Objective:To discuss the effect of knock down of gene of kinesin family member 3B(KIF3B),an important component of primary cilia(PC)of in mouse embryonic palatal mesenchymal on the autophagy level of cells(mEPMCs)cells,and to clarify its mechanism.Methods:The mEPMCs from gestational day 14.5 C57BL/6J mice cultured in vitro were collected and divided into control group(administered normal saline),empty lentivirus transfected cell group(sh-NC group)(administered lentivirus transfection),KIF3B knockdown group(sh-KIF3B group)(administered KIF3B gene knockdown),and KIF3B knockdown plus Smoothened receptor agonist(SAG)group(sh-KIF3B+SAG group)(administered KIF3B gene knockdown followed by SAG addition),based on whether the KIF3B gene was knocked down and whether the SAG was used to activate the sonic hedgehog(Shh)signaling pathway and its downstream coreceptor Smo,with 5 rats in each group.Transmission electron microscope was used to observe the morphology and the number of autophagosomes/autolysosomes in the mEPMCs in various groups;Western blotting method was used to detect the expression levels of autophagy-related proteins Beclin-1 and p62,and the Shh signaling pathway proteins Shh and Smo in the mEPMCs in various groups.Results:The transmission electron microscope observation results showed that compared with control group,the number of autophagosomes/autolysosomes in sh-KIF3B group was significantly increased(P<0.05);compared with sh-KIF3B group,the number of autophagosomes/autolysosomes in the mEPMCs in sh-KIF3B+SAG group was significantly decreased(P<0.05).The Western blotting results showed that compared with control group,the Beclin-1 protein expression level in the mEPMCs in sh-KIF3B group was significantly increased(P<0.05),and the KIF3B,p62,Shh,and Smo protein expression levels were significantly decreased(P<0.01);compared with sh-KIF3B group,the Shh,Smo,and p62 protein expression levels in the mEPMCs in sh-KIF3B+SAG group were significantly increased(P<0.01),and the Beclin-1 protein expression level was significantly decreased(P<0.01).Conclusion:Knockdown of KIF3B gene can promote autophagy of the mEPMCs,and the mechanism may be related to its inhibition of the Shh signaling pathway.

Liver transplantation (LT) has become a standard treatment for end-stage liver diseases, and graft injury is intricately associated with poor prognosis. Granzyme B (GZMB) plays a vital role in natural killer (NK) cell biology, but whether NK-derived GZMB affects graft injury remains elusive. Through the analysis of single-cell RNA-sequencing data obtained from human LT grafts and the isolation of lymphocytes from mouse livers following ischemia-reperfusion injury (IRI), we demonstrated that 2NK cells with high expression of GZMB are enriched in patients and mice. Both systemically and liver-targeted depletion of NK cells led to a notable reduction in GZMB⁺ cell infiltration, subsequently resulting in diminished graft injury. Notably, the reconstitution of Il2rg ^-/- Rag2 ^-/- mice with purified Gzmb-KO NK cells demonstrated superior outcomes compared to those with wild-type NK cells. Crucially, global knockout of GZMB and pharmacological inhibition exhibited remarkable improvements in liver function in both mouse IRI and rat LT models. Moreover, a phosphorylated derivative of FDA-approved vidarabine was identified as an effective inhibitor of mouse GZMB activity by molecular dynamics, which could provide a potential avenue for therapeutic intervention. Therefore, targeting NK cell-derived GZMB during the LT process suggests potential therapeutic strategies to improve post-transplant outcomes.

Objective:To investigate the key genetic mutations during the progressive disease (PD) /leukemic transformation (LT) course in MDS by analyzing the dynamic changes of genetic mutations in patients with myelodysplastic neoplasms (MDS) with or without PD/LT.Methods:This study enrolled 84 patients with sequential MDS from May 2019 to August 2023 at ZhongDa Hospital Southeast University and used the next generation sequencing to detect gene mutations. The dynamic changes of genetic mutations in patients with MDS with or without PD/LT were retrospectively analyzed.Results:①This study analyzed data from 84 patients diagnosed with MDS with a median age of 63 (range: 31-95) years and consisting of 51 males and 33 females. Participants were distributed to the PD cohort ( n=20), LT cohort ( n=13), and non-PD/LT cohort ( n=51). Patients from the PD/LT cohorts demonstrated a higher proportion of bone marrow blasts than the non-PD/LT cohort at the first sequencing (1.6% vs. 0.4%, P=0.013). ②The most frequently mutated genes that were detected at first sequencing were ASXL1 ( n=21, 25.0%), TP53 ( n=17, 20.2%), TET2 ( n=12, 14.3%), DNMT3A ( n=11, 13.1%), and U2AF1 ( n=11, 13.1%). Further, patients from the PD/LT cohorts exhibited a higher median number of mutated genes than the non-PD/LT cohort (2 vs.1, P=0.014) at first sequencing. TET2 (27.3% vs. 5.9%, P=0.010), SETBP1 (15.2% vs.2.0%, P=0.033), and RUNX1 (18.2% vs. 2.0%, P=0.013) mutations were enriched in the PD/LT cohorts than in the non-PD/LT cohort. ③The most frequently detected acquired mutations (Ⅰ mutations) and clonally expanded mutations (Ⅱ mutations) were TP53 ( n=9, 10.7%), TET2 ( n=7, 8.3%), ASXL1 ( n=7, 8.3%), and RAS pathway ( n=7, 8.3%). Furthermore, patients from the PD/LT cohorts showed a higher median number of Ⅰ/Ⅱ genes than the non-PD/LT cohort (2 vs. 0, P<0.001), and Ⅰ/Ⅱ RAS pathway (21.2% vs. 0, P=0.001), TP53 (27.3% vs. 0, P<0.001), and TET2 (18.2% vs. 2.0%, P=0.013) mutations were enriched in PD/LT cohorts than in the non-PD/LT cohorts. ④Most of the TP53 mutations (9/12, 75.0%) in PD/LT cohorts were Ⅰ/Ⅱ mutations, whereas all of the TP53 mutations in non-PD/LT cohort were clone-decrease mutations (Ⅲ mutations) (5/8, 62.5%) or clone-stable mutations (Ⅳ mutations) (3/8, 37.5%). Most of the RAS pathway mutations (7/8,87.5%) in the PD/LT cohorts were Ⅰ/Ⅱ mutations, whereas only one patient in the non-PD/LT cohort demonstrated RAS pathway mutations, which belonged to Ⅳ mutations. Conclusion:Patients from the PD/LT cohorts demonstrated a higher proportion of bone marrow blasts and a higher median number of mutations than the non-PD/LT cohort at first sequencing; TET2, SETBP1, and RUNX1 mutations were enriched in the PD/LT cohorts than in the non-PD/LT cohort at first sequencing. Patients from the PD/LT cohorts exhibited a higher number of Ⅰ/Ⅱ mutations than the non-PD/LT cohort. Further, Ⅰ/Ⅱ TP53, RAS pathway, and TET2 mutations were enriched in the PD/LT cohorts, and Ⅰ/Ⅱ TP53 and RAS pathway mutations may contribute to the PD/LT.

Metachromatic leukodystrophy (MLD) is an inherited disease caused by a deficiency of the enzyme arylsulfatase A (ARSA). Lentivirus-modified autologous hematopoietic stem cell gene therapy (HSCGT) has recently been approved for clinical use in pre and early symptomatic children with MLD to increase ARSA activity. Unfortunately, this advanced therapy is not available for most patients with MLD who have progressed to more advanced symptomatic stages at diagnosis. Patients with late-onset juvenile MLD typically present with a slower neurological progression of symptoms and represent a significant burden to the economy and healthcare system, whereas those with early onset infantile MLD die within a few years of symptom onset. We conducted a pilot study to determine the safety and benefit of HSCGT in patients with postsymptomatic juvenile MLD and report preliminary results. The safety profile of HSCGT was favorable in this long-term follow-up over 9 years. The most common adverse events (AEs) within 2 months of HSCGT were related to busulfan conditioning, and all AEs resolved. No HSCGT-related AEs and no evidence of distorted hematopoietic differentiation during long-term follow-up for up to 9.6 years. Importantly, to date, patients have maintained remarkably improved ARSA activity with a stable disease state, including increased Functional Independence Measure (FIM) score and decreased magnetic resonance imaging (MRI) lesion score. This long-term follow-up pilot study suggests that HSCGT is safe and provides clinical benefit to patients with postsymptomatic juvenile MLD.
Humans ; Leukodystrophy, Metachromatic/genetics* ; Pilot Projects ; Genetic Therapy/methods* ; Hematopoietic Stem Cell Transplantation ; Male ; Follow-Up Studies ; Female ; Lentivirus/genetics* ; Child ; Child, Preschool ; Hematopoietic Stem Cells/metabolism* ; Cerebroside-Sulfatase/metabolism* ; Adolescent

Vacuum compression molding (VCM) is a novel technology supporting the research and development of pharmaceutical solid dispersions. It is widely applied due to its precision and convenience in sample preparation. This technology integrates the principles of heating, melting, cooling, and vacuum compression to transform solid powders into shaped solids directly. By selecting different molds, temperatures, and pressures, researchers can prepare samples with diverse characteristics. This paper presents an overview of the equipment composition and working principles of VCM technology, demonstrating its distinct advantages in the formulation screening process of amorphous solid dispersions through comparative analysis with hot melt extrusion using case studies, and introduces its applications in the development of drug delivery systems and rheological characterization analysis, with a perspective on the future development of its functions.