The core pathogenesis of attention deficit hyperactivity disorder （ADHD） lies in deficiency， stasis and stagnation. Deficiency arises from kidney essence depletion and spleen dysfunction in transportation and transformation， leading to inadequate nourishment of the marrow sea. Stasis caused by qi deficiency leads to obstruction in channels and collaterals， resulting in obstructed marrow transport. Stagnation is associated with the excess of the five minds transforming into fire， which scorches the brain orifices and leads to loss of control over marrow utilisation. Based on this， a "supplementation-unblocking-regulation" therapeutic approach is proposed. For deficiency， the focus is on supplementing kidney and fortifying spleen， and replenishing the marrow sea. For stasis， the priority is to unblock and open the orifices， and clear the marrow channels. For stagnation， the core is to clear fire and contain the mind， regulate and restore vital activity. In clinical practice， it is necessary to identify the primary and secondary pathogenic mechanisms and apply dynamic， combined treatment， integrating Chinese herbal medicine， acupuncture， and guiding exercises throughout the process， aiming to provide a reference for the diagnosis and treatment of ADHD with traditional Chinese medical.

Attention deficit hyperactivity disorder （ADHD） is often accompanied by tic disorder. The core pathogenesis is considered to be ministerial fire scorching yin and internal stirring of deficient wind， which leads to disharmony between the body and spirit， resulting in clinical manifestations. The treatment principles emphasize nourishing yin fluids， calming ministerial fire， and extinguishing endogenous wind （内风）. The method of nourishing yin fluids is applied throughout the entire treatment process， commonly using ingredients such as Shudihuang （Rehmanniae Radix Praeparata）， Shanzhuyu （Corni Fructus）， Gouqizi （Lycii Fructus）， Wuweizi （Schisandrae Chinensis Fructus）， and Tusizi （Cuscutae Semen）. These are combined with approaches to harmonize the zang-fu organs， primarily including extinguishing liver wind， clearing heart fire， nourishing kidney water， and strengthening spleen earth， thereby stabilizing ministerial fire and extinguishing endogenous wind. Additionally， emotional regulation and smoothing emotional constraint are essential to improve clinical symptoms in children with ADHD comorbid with tic disorder.

OBJECTIVE To establish the high-performance liquid chromatography （HPLC） fingerprint of Sagina japonica ， and to establish a quantitative analysis of multi-components by single-marker （QAMS） method for simultaneous determination of six componen ts in S. japonica ， aiming to provide references for the quality control of this medicinal herb. METHODS HPLC method was used to establish the fingerprints of 12 batches （No. S1-S12） of S . japonica according to Similarity Evaluation System of Chromatographic Fingerprint of Traditional Chinese Medicine . The similarity evaluation and identification of common peaks were conducted， followed by cluster analysis （CA） and principal component analysis （PCA） for 12 batches of samples. Using vicenin-2 as internal reference， the contents of p-hydroxy cinnamic acid， apigenin-6-C-arabinoside-8-C-glucoside， isoorientin， vitexin and 20-hydroxyecdysone were determined by QAMS method. The results were then compared with those obtained by the external standard method. RESULTS The similarities of HPLC fingerprints for 12 batches of S . japonica ranged from 0.828-0.998. A total of 17 common peaks were calibrated， and 6 common peaks were identified. Specifically， peak 5 was identified as vicenin-2， peak 7 as p-hydroxycinnamic acid， peak 10 as apigenin-6-C-arabinoside-8-C-glucoside， peak 11 as isoorientin， peak 13 as vitexin， and peak 15 as 20-hydroxyecdysone. The results of CA showed that S1-S5， S7 and S9-S11 were clustered into one category， S6 was clustered into one category， and S8 and S12 were clustered into one category. The results of PCA revealed that the accumulative contribution rate of the four main components was 89.430%. The content ranges measured by QAMS method for p-hydroxy cinnamic acid， apigenin-6-C-arabinoside-8-C-glucoside， isoorientin， vitexin and 20-hydroxyecdysone were 0.017 4-0.269 4， 0.568 8-4.240 3， 0.503 2-5.040 3， 0.024 0-0.132 0 and 2.551 3-4.881 1 mg/g， respectively. There was no significant difference in the contents of components measured between QAMS method and the external standard method （ P ＞0.05）. CONCLUSIONS The established HPLC fingerprint and QAMS method can be used for quality evaluation and quality control of S . japonica.

Objective:To investigate the function and mechanism of exosome Linc00665 in modulating CD8+T cell immunoreactivity to promote radiotherapy resistance in OSCC.Methods:HOEC,SCC9 and SCC9-RR exosomes were extracted and identified,and the expression of Linc00665 was detected by qRT-PCR in cell lines and exosomes.The expression of TNF-α,IFN-γ,perforin and granzyme B in each treatment group was detected by ELISA(PBS,SCC9 exo,SCC9-RR exo).The killing ability of CD8+T cells against SCC9 cells in each treatment group was detected by CCK-8 assay.The targets of Linc00665 were further bioinformatically ana-lyzed and verified by qRT-PCR and Western blot.The expression of Linc00665,miR-28-5p and PD-1 in CD8+T cells was exogenous-ly regulated,the expression of immunoreactive factors in the supernatants of each treatment group was detected by ELISA(NC,sh-Linc00665,miR-28-5p inhibitor,sh-PD-1),and the killing ability of cells in each group was detected by CCK-8 method.Results:The concentrations of TNF-α,IFN-γ,perforin and granzyme B in the supernatants of cell culture in the SCC9-RR exo/CD8+T group were significantly decreased compared with those in the PBS/CD8+T group and the SCC9 exo/CD8+T group(P＜0.05),and the kill-ing ability of the cells in the SCC9-RR exo group was significantly decreased compared with those in the PBS group and the SCC9 exo group(P＜0.05),suggesting that SCC9-RR exo could inhibit the tumor killing ability of CD8+T cells.qRT-PCR results suggested that Linc00665 was highly expressed in the SCC9-RR cell line as well as exosome(P＜0.05).It was further verified by bioinformat-ics analysis that Linc00665 could regulate PD-1 expression via miR-28-5p,thereby modulating CD8+T cell immunoreactivity to pro-mote OSCC radiotherapy resistance.Conclusion:Exosome Linc00665 regulates CD8+T cell immunoreactivity through miR-28-5p/PD-1 axis to promote OSCC radiotherapy resistance.

Objective To investigate the predicting value of cardiac magnetic resonance feature tracking(CMR-FT)for adverse left ventricular remodeling(ALVR)in patients with acute anterior wall ST-segment elevation myocardial infarction(STEMI).Methods The clinical data and cardiac magnetic resonance(CMR)images of 161 acute anterior wall STEMI patients within 1 week and 6 months after emergency percutaneous coronary intervention(PCI)were retrospectively analyzed.ALVR was defined as an increase of left ventricular end-diastolic volume(LVEDV)over 20％at the second CMR examination compared to the baseline.The CMR parame-ters were analyzed by CVI42 post-processing software.The logistic regression analysis was used to screen the independent predictors of ALVR,and the receiver operating characteristic(ROC)curve was used to evaluate the predictive efficiency of ALVR.Results The incidence of ALVR at 6 months was 21.7％(35/161).The logistic regression analysis showed that the left ventricular global circumferential strain(LVGCS)and right ventricular global longitudinal strain(RVGLS)at baseline were independent predictors for ALVR(P＜0.001).When LVGCS was-13.89％and RVGLS was-15.07％at baseline,the sensitivity of predicting ALVR was 0.714 and 0.743,the specificity was 0.833 and 0.810,and the area under the curve(AUC)was 0.806 and 0.835,respectively.The sensitivity of LVGCS combined with RVGLS in predicting ALVR was 0.802,the specificity was 0.952,and the AUC was 0.888.The DeLong test showed that the AUC of LVGCS com-bined with RVGLS in predicting ALVR was significantly higher than that of individuals,and the difference was statistically significant(P＜0.05).Conclusion The LVGCS and RVGLS at baseline are independent predictors for ALVR in patients with acute anterior wall STEMI,their combination can significantly improve the pre-dictive efficiency of ALVR in these patients.

Objective:To study the influence of methyltransferases like 3(METTL3)on the radiosensitivity of oral squamous cell carcinoma cells(OSCCs).Methods:The apoptosis level of OSCCs CAL27,SCC9 and SCC15 treated with X-ray radiation doses of 2,4 and 8 Gy respectively was compared by flow cytometry,the expression of methylated gene RNA and protein in the cells were examined by qRT-PCR and Western blot.m6A in the cells was quantified by LC/LC-MS method.qRT-PCR and Western blot were used to investigate the expression of methylated gene RNA and protein in the cells.Flow cytometry was used to examine the cell apoptosis level of CAL27 and SCC15 cells treated with METTL3 overexpression and knockdown respectively.The clone forma-tion of CAL27 and SCC15 cells after knockdown and overexpression of target genes followed by radiation treatment was observed by clonogenic assays.Results:The apoptosis rate of all the cell lines increased with the increase dose of radiation at each dose,CAL27 cells showed the highest and SCC15 showed the lowerst apoptosis rate.The RNA and protein expression levels of METTL3 in CAL27 were significantly lower than those of SCC15.m6A quantification showed that the methylation modification in CAL27 cells was lower than that in SCC15.The expression of METTL3 was increased in CAL27 and SCC15 cells after radiation treatment.Knockdown of METTL3 increaced the apoptosis rate and decreased the clonogenesity,overession of METTL3 the decreaced the ap optosis rate and increased the clonogenecity of the cells.Conclusion:Regulation of METTL3 can affect the radiotherapy sensitivity of OSCCs,METTL3 may become a new target for radiosensitization of OSCCs.

Objective:To explore the clinical phenotype and genetic characteristics of a child with hereditary Spastic paraplegia type 52 (SPG52) due to variant of AP4S1 gene. Methods:A child diagnosed with SPG52 at the Department of Pediatrics of the First Affiliated Hospital of Anhui Medical University in May 2010 was selected as the study subject. Whole-exome sequencing (WES) was carried out for the child and his parents. Candidate variants were confirmed by Sanger sequencing. Pathogenicity of the candidate variant was interpreted according to the guidelines from the American College of Medical Genetics and Genomics (ACMG). The study protocol was approved by the Ethics Committee of the Hospital (Ethics No.: PJ2024-04-56).Results:The child had presented with global developmental delay from infancy, and featured progressive lower limb spasticity, contractures, talipes equinovarus, and muscle weakness, but with no significant facial dysmorphism. His first febrile seizure occurred before one year of age, followed by several afebrile seizures. The seizures had remitted after 3 to 4 years of antiepileptic therapy, and electroencephalography was normal. However, he had severe intellectual disability, and MRI revealed reduced white matter. WES identified a homozygous AP4S1 c. 289C>T (p.Arg97*) variant in the child, for which both of his parents were heterozygous carriers. The variant was rated as pathogenic based on the ACMG guidelines. Literature review has identified 8 publications on SPG52, involving 18 patients from 12 pedigrees. Combined with our case, 14 had carried homozygous variants of the AP4S1 gene, 3 had compound heterozygous variants, and 2 had heterozygous variants, involving 12 distinct variant sites. The cohort included 7 males and 12 females. All patients exhibited progressive lower limb spasticity and weakness as the primary feature, with certain loss of independent ambulation. Most patients had intellectual disability, some had distinctive facial features, though febrile seizures or epilepsy were common. Electroencephalography often showed increased slow-wave activity. Brain MRI frequently demonstrated ventriculomegaly, a thin corpus callosum, and reduced white matter. Conclusion:The homozygous c. 289C>T (p.Arg97*) variant of the AP4S1 gene probably underlay the pathogenesis of SPG52 in this child. Above discovery has expanded the mutational spectrum of AP4S1 and provided valuable insights for the genetic diagnosis, counseling, and clinical management of SPG52.

Objective To explore the impact of Metformin(Met)on the balance of type 17 helper T cells(Th17)/regulatory T cells(Treg)in DM rats by regulating the C-C motif chemokine ligand 2(CCL2)-C-C motif chemokine receptor 2(CCR2)axis.Methods A total of 50 SD rats were randomly divided into normal control(NC)group,model(Mod)group,Met(100 mg/kg)group,Met+empty vector(Met+oe-NC)group,and Met+CCL2 overexpression(Met+oe-CCL2)group,with 10 rats in each group.The changes of FBG evaluated in each group,and the pathological changes of pancreatic islet tissue were assessed using hematoxylin-eosin(HE)staining followed by histological scoring.The ratio of Th17/Treg in peripheral blood was detected by flow cytometry.ELISA was used to measure the serum levels of IL-17,TNF-α,TGF-β and IL-10.Immunofluorescence was used to detect the proportion of Th17 and Treg in pancreatic tissue.RT-PCR and Western blot were used to detect the mRNA and protein expression of ROR-γt,forkhead box P3(FOXP3),CCL2 and CCR2 in pancreatic tissue.Results The islet structure was intact and the boundary was clear,and there was no pathological damage in the NC group.In the Mod group,the islet tissue exhibited significant pathological damage,characterized by disrupted architecture,reduced volume,and blurred boundaries with surrounding tissues.Additionally,the cells in the islet displayed marked disorganization and substantial loss.Compared with the Mod group,the pathological damage of the islet tissue was alleviated in the Met and Met+oe-NC group.Compared with the Met group,the pathological damage was aggravated in the Met+oe-CCL2 group.Compared with the NC group,the levels of FBG,Th17/Treg ratio,IL-17,TNF-α,Th17 infiltration rate,ROR-γt,CCL2,and CCR2 mRNA and protein expression were higher(P<0.05),while TGF-β,IL-10,Treg infiltration rate,FOXP3 mRNA and protein expression were lower in the Mod group(P<0.05).Compared with the Mod group,the levels of FBG,Th17/Treg ratio,IL-17,TNF-α,Th17 infiltration rate,ROR-γt,CCL2,and CCR2 mRNA and protein were lower(P<0.05),while TGF-β,IL-10,Treg infiltration rate,FOXP3 mRNA and protein expression were higher in the Met and Met+oe-NC groups(P<0.05).Compared with the Met group,the levels of FBG,Th17/Treg ratio,IL-17,TNF-α,Th17 infiltration rate,ROR-γt,CCL2,and CCR2 mRNA and protein were higher(P<0.05),and TGF-β,IL-10,Treg infiltration rate,FOXP3 mRNA and protein expression were higher in the Met+oe-CCL2 group(P<0.05).Conclusions Metformin can promote Th17/Treg cell balance drift towards Treg direction by inhibiting the activation of CCL2-CCR2 signal axis,thereby preventing the occurrence and development of inflammatory reactions,alleviating pancreatic islet damage,and improving the glucose metabolism in DM rats.

Objective:To investigate the effects of sampling methods on pathological assessment of resected non-small cell lung cancer (NSCLC) specimen with tumor maximum diameter >3 cm after neoadjuvant therapy.Methods:NSCLC patients with a large tumor (diameter >3 cm) that were resected after neoadjuvant therapy from June 2020 to July 2023 were retrospectively collected in the Department of Pathology, Shanghai Pulmonary Hospital, Shanghai, China. Sampling methods of the tumor bed were performed in accordance with the international and Chinese experts recommendations for resection specimens following neoadjuvant therapy (recommended sampling method, RSM), and all remaining tumor bed lesions were completely sampled after recommended sampling (complete sampling method, CSM). The difference of pathological response assessment of residual viable tumor (RVT) between RSM and CSM was examined.Results:A total of 90 cases were identified and analyzed, including 39 cases of squamous cell carcinoma and 51 cases of adenocarcinoma, treated with neoadjuvant therapy including chemotherapy in 22 cases (24.4%), targeted therapy in 14 cases (15.6%), and chemoimmunotherapy in 54 cases (60.0%). There were 62 males and 28 females with an average age of (62.7±17.9) years. The average tumor maximum diameter was 4.3 cm (range, 3.1-8.0 cm). The average number of sampled blocks was 8 blocks (range, 5 to 16) and 15 blocks (range, 8 to 36) per case by RSM and CSM, respectively. According to the definition of major pathological response (MPR) in which RVT is ≤10%, the numbers of patients with MPR were 34 cases by RSM and 30 cases by CSM, respectively. Four cases showed inconsistent RVT between the two methods, including one case of squamous cell carcinoma and three cases of adenocarcinoma. The RVT of the four inconsistent cases was 7%, 7%, 5% and 9% (MPR by RSM), and 15%, 15%, 15% and 20% (non-MPR by CSM), respectively. The kappa values of MPR consistency evaluated by the two sampling methods were 0.893 for all cases, 0.906 for squamous cell carcinoma cases and 0.751 for adenocarcinoma cases. According to MPR cut-off of 65% for invasive primary adenocarcinoma, 24 cases and 20 cases achieved MPR by RSM and CSM, respectively. Of the four inconsistent cases, the RVT by RSM was 60% in three cases and 65% in one case (MPR), whereas the RVT by CSM was 70% in three cases and 75% in one case (non-MPR). The kappa value of the two sampling methods was 0.741.Conclusions:There is high consistency between RSM and CSM in the pathological assessment of post-treatment responses in resected NSCLC specimens with tumor maximum diameter larger than 3 cm. When the percentage of RVT cells is close to MPR, re-evaluation of the specimen is required and CSM may be necessary to accurately evaluate the degree of pathological remission, assist in clinical postoperative treatment, and predict patient prognosis.

Objective To explore the impact of Metformin(Met)on the balance of type 17 helper T cells(Th17)/regulatory T cells(Treg)in DM rats by regulating the C-C motif chemokine ligand 2(CCL2)-C-C motif chemokine receptor 2(CCR2)axis.Methods A total of 50 SD rats were randomly divided into normal control(NC)group,model(Mod)group,Met(100 mg/kg)group,Met+empty vector(Met+oe-NC)group,and Met+CCL2 overexpression(Met+oe-CCL2)group,with 10 rats in each group.The changes of FBG evaluated in each group,and the pathological changes of pancreatic islet tissue were assessed using hematoxylin-eosin(HE)staining followed by histological scoring.The ratio of Th17/Treg in peripheral blood was detected by flow cytometry.ELISA was used to measure the serum levels of IL-17,TNF-α,TGF-β and IL-10.Immunofluorescence was used to detect the proportion of Th17 and Treg in pancreatic tissue.RT-PCR and Western blot were used to detect the mRNA and protein expression of ROR-γt,forkhead box P3(FOXP3),CCL2 and CCR2 in pancreatic tissue.Results The islet structure was intact and the boundary was clear,and there was no pathological damage in the NC group.In the Mod group,the islet tissue exhibited significant pathological damage,characterized by disrupted architecture,reduced volume,and blurred boundaries with surrounding tissues.Additionally,the cells in the islet displayed marked disorganization and substantial loss.Compared with the Mod group,the pathological damage of the islet tissue was alleviated in the Met and Met+oe-NC group.Compared with the Met group,the pathological damage was aggravated in the Met+oe-CCL2 group.Compared with the NC group,the levels of FBG,Th17/Treg ratio,IL-17,TNF-α,Th17 infiltration rate,ROR-γt,CCL2,and CCR2 mRNA and protein expression were higher(P<0.05),while TGF-β,IL-10,Treg infiltration rate,FOXP3 mRNA and protein expression were lower in the Mod group(P<0.05).Compared with the Mod group,the levels of FBG,Th17/Treg ratio,IL-17,TNF-α,Th17 infiltration rate,ROR-γt,CCL2,and CCR2 mRNA and protein were lower(P<0.05),while TGF-β,IL-10,Treg infiltration rate,FOXP3 mRNA and protein expression were higher in the Met and Met+oe-NC groups(P<0.05).Compared with the Met group,the levels of FBG,Th17/Treg ratio,IL-17,TNF-α,Th17 infiltration rate,ROR-γt,CCL2,and CCR2 mRNA and protein were higher(P<0.05),and TGF-β,IL-10,Treg infiltration rate,FOXP3 mRNA and protein expression were higher in the Met+oe-CCL2 group(P<0.05).Conclusions Metformin can promote Th17/Treg cell balance drift towards Treg direction by inhibiting the activation of CCL2-CCR2 signal axis,thereby preventing the occurrence and development of inflammatory reactions,alleviating pancreatic islet damage,and improving the glucose metabolism in DM rats.