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.

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.

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.

Objective:To investigate the nutritional risk status of children in PICU and analyze its influencing factors.Methods:From July 2021 to February 2023,all children aged 1 to 18 years admitted to PICU of Beijing Children's Hospital were investigated by using the pediatric Yorkhill Malnutrition Scoring tool(PYMS)and the clinical data questionnaire.Results:A total of 492 children in PICU were enrolled.The first nutritional risk screening results showed that there were 32 cases of no/low nutritional risk(6.5%),76 cases of medium risk(15.4%),and 384 cases of high risk(78.1%).The incidence of medium/high nutritional risk was as high as 93.5%.The PYMS score of nutritional risk in PICU was(2.61±1.42).The results of multiple linear regression analysis showed that weight,fever time before admission,white blood cells,body mass index,primary diagnosis,father's education,and diet before illness were the main influencing factors of nutritional risk of children in PICU(P<0.05).Conclusion:Children in PICU are in a state of high nutritional risk.It is suggested that children in PICU should carry out nutritional screening in a standardized manner,identify children with high nutritional risk and its influencing factors early.To actively conduct nutritional assessment and nutritional intervention could improve the clinical outcome of children in PICU.

Objective:To explore the mechanism of Shenfu Injection in treating chronic heart failure (CHF) based on Fas/FasL apoptosis signaling pathway.Methods:A total of 70 SPF male C57BL/6 mice were divided into blank group (15 mice) and model group (55 mice) according to random number table. The CHF model was prepared by intraperitoneal injection of isoproterenol. After 4 weeks, the successfully modeled mice were randomly divided intoa model group, Shenfu Injection group, and Western medicine group using a random number table method. After adfministration for 15 d, the left ventricular ejection fraction (LVEF) and left ventricular shortening fraction (LVFS) of each group of mice were measured by heart color ultrasound 1; serum NT-proBNP was detected by enzyme-linked immunosorbent assay (ELISA); Hematoxylin-eosin staining (HE) was used to observe the changes of myocardial tissue morphology; TUNEL pod method was used to detect apoptosis; the mRNA transcription levels of tumor necrosis factor-α (TNF-α), Fas, Fas ligand (FasL), Caspase-3 and Caspase-8 were detected by RT-PCR; protein expression levels of TNF-α, Fas, FasL, Caspase-3 and Caspase-8 in myocardial tissue were detected by Western blot. The logarithmically grown H9c2 cells were divided into blank group, model group and Shenfu Injection group. The cells in blank group were cultured for 48 hours; cells in the model group were exposed to 80 μmol/L isoproterenol for 48 hours; cells in Shenfu Injection group were pretreated with 5 μl/ml Shenfu injection for 3 hours and exposed to 80 μmol/L isoproterenol for 48 h. The cell growth was observed under microscope. Western blot and RT-PCR were used to detect the expression of related proteins and mRNA transcription. TUNEL pod and flow cytometry were used to detect apoptosis.Results:Compared with the model group, the cardiac function improved in the model group and Shenfu Injection group, serum NT ProBNP levels decreased ( P<0.01 or P<0.05), myocardial cell injury and apoptosis were reduced ( P<0.01 or P<0.05), and the mRNA and protein expression of TNF-α, Fas, FasL, Caspase-3, Caspase-8 in myocardial tissue decreased ( P<0.01 or P<0.05). The H9c2 cells in the blank group were spindle shaped with clear structure; the myocardial cells in the model group shrank and became shorter, with blurred cell boundaries and exhibiting apoptotic and necrotic morphology; Most H9c2 cells in the reference group were spindle shaped, with reduced cell death and increased density. Compared with the model group, the levels of TNF-α, Fas, FasL, Caspase-3, Caspase-8 protein and mRNA in H9c2 cells of the Shenfu Injection group decreased ( P<0.01 or P<0.05), and the apoptosis rate decreased ( P<0.01). Conclusion:Shenfu Injection may improve cardiac function and cardiomyocyte apoptosis of CHF mice through Fas/FasL signaling pathway.

Objective:To present a set of clinically representative quality assurance (QA) test cases for stereotactic radiosurgery (SRT) plans of multiple intracranial metastases, in order to assess the plan quality and machine execution capabilities.Methods:Based on the clinical characteristics of multiple brain metastases, four groups of test cases with three target volumes (TVs), six TVs, nine TVs, and TVs near organs at risk (OARs) were designed. For these cases, SRT plans were developed, and plan quality was assessed using metrics including the Radiation Therapy Oncology Group conformality index (RTOG CI), gradient index (GI), homogeneity index (HI), and the volume of normal brain tissue receiving a dose of 24 Gy ( V24 Gy), which was defined as the volume enclosed by the 24 Gy isodose line around the Brain-PTV ( V24 Gy of Brain-PTV). Verification plans were generated for each test case, including the verification of point doses, planar doses (PD), and SRS MapCHECK (SMC) semiconductor matrix planar doses. Compared with the calculated result of the treatment planning system (TPS), the criteria for the γ analysis of planar doses were set at 1 mm/2% and 2 mm/2%. Results:For the four groups of test cases, the mean CI, GI, HI, and V24 Gy of Brain-PTV were 1.04±0.03, 3.79±0.40, 0.73±0.01 and (7.46±3.80) cm 3, respectively. The mean deviations of the point doses were 0.88%±0.98%, 1.47%±0.79%, 1.52%± 0.76%, and 1.17% ± 0.38%, respectively. The mean γ passing rates of the single fields for PDs were greater than 98% at 2 mm/2% and exceeding 96% at 1 mm/2%, and the mean γ pass rates of the SMC semiconductor matrix for PDs were 97.75% ± 2.31% and 99.33% ± 0.62%, at 1 mm/2% and 2 mm/2% respectively. Conclusions:The proposed QA test cases for SRT of multiple intracranial metastases allow for the effective assessments of the plan quality and machine execution capabilities and, thus, can assist various centers in clinical applications.

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.

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.

Aim To investigate the effect of Panax no-toginseng saponins(PNS)on the interaction between endometrial cancer cells and macrophages by regulating the epidermal growth factor receptor(EGFR)/heat shock protein 27(HSP27)axis.Methods Ishikawa cells were divided into Control,DDP,PNS-treated,M2-CM,M2-CM+DDP,M2-CM+PNS,M2-CM+PNS+oe-NC,M2-CM+PNS+oe-EGFR,PNS(200 mg·L-1)+oe-NC,PNS(200 mg·L-1)+oe-EG-FR,oe-NC,oe-EGFR,oe-EGFR+si-NC,and oe-EG-FR+si-HSP27 groups.MTT assay was used to detect cell proliferation,Transwell assay for cell invasion,TUNEL assay for cell apoptosis,qRT-PCR for macro-phage polarization markers CD86 and CD163 mRNA levels,ELISA for iNOS and IL-12 levels,and West-ern-blot for EGFR and HSP27 protein expression.A nude mouse xenograft tumor model was established and treated with PNS to evaluate the effect of PNS in vivo.Results Compared with the Control group,PNS and DDP significantly inhibited the proliferation and inva-sion of Ishikawa cells and induced apoptosis.M2-CM treatment inhibited M1 macrophage markers,promoted M2 macrophage markers,and induced Ishikawa cell growth,but this effect was reversed by PNS treatment.EGFR was confirmed as a target of PNS,and compared with the M2-CM+PNS+oe-NC group,EGFR overex-pression promoted M2 macrophage marker levels,in-duced Ishikawa cell proliferation and invasion,and in-hibited apoptosis.Knockdown of HSP27 reversed the effect of EGFR overexpression on the biological behav-ior of Ishikawa cells.Animal experiments showed that PNS could inhibit tumor growth and reduce the positive expression of CD163,EGFR,and HSP27 in tumor tis-sues.Conclusion PNS affects the interaction be-tween macrophages and EC cells by regulating the EG-FR/HSP27 axis,thereby participating in EC progres-sion.