OBJECTIVE To provide a scientific basis for the quality evaluation and clinical application of Qingwen hufei granules. METHODS Fourteen batches of Qingwen hufei granules were used as samples to establish high-performance liquid chromatography （HPLC） fingerprints using the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine （2012 Edition）. The chromatographic peaks were identified and the similarity was evaluated. Cluster analysis （CA）， principal component analysis （PCA）， and orthogonal partial least squares-discriminant analysis （OPLS-DA） were used to conduct chemical pattern recognition analysis on the 14 batches of samples. Meanwhile， the contents of neochlorogenic acid （NGA）， chlorogenic acid （CHA）， cryptochlorogenic acid （CGA）， forsythoside A （FTA）， 3，5-O-dicaffeoylquinic acid （3，5-O- DA）， 4，5-O-dicaffeoylquinic acid （4，5-O-DA）， and angoroside C （AGC） in the samples were determined by HPLC. RESULTS The methodological investigation results of both the fingerprint and the content determination complied with the relevant requirements. Fourteen common peaks were indicated in the HPLC fingerprints of the 14 batches of samples， and 7 of them were identified [NGA （peak 2）， CHA （peak 3）， CGA （peak 5）， FTA （peak 11）， 3，5-O-DA （peak 12）， 4，5-O-DA （peak 13）， and AGC （peak 14）]； the similarity of each sample was greater than 0.94. The results of CA and PCA showed that the samples could be classified into 3 categories； the results of OPLS-DA indicated that peak 4 （unknown）， peak 11 （FTA）， peak 8 （unknown）， peak 9 （unknown）， and peak 1 （unknown） were the differential components. The content ranges of NGA， CHA， CGA， 3，5-O-DA， FTA， 4，5-O-DA and AGC in the 14 batches of samples were 0.210 4-0.458 7， 0.269 1-0.506 3， 0.228 1-0.461 1， 0.443 9-1.044 6， 0.066 7-0.155 7， 0.062 8-0.143 8， and 0.057 4-0.105 7 mg/g， respectively. CONCLUSIONS The HPLC fingerprint and multi-component content determination methods established in this study are efficient and reliable， and can be used for the quality evaluation of Qingwen hufei granules.

Objective: To investigate the damaging effects of red blood cell supernatant (RBC-S) stored for different durations (7 d, 14 d, and 28 d) on vascular endothelial cells, and to explore the underlying mechanisms using bioinformatics analysis, so as to provide references for optimizing red blood cell transfusion strategies. Methods: Human umbilical vein endothelial cells (HUVECs) were co-cultured with RBC-S stored for 7, 14 and 28 days, designated as the 7 d group, 14 d group and 28 d group respectively, which were collectively defined as the experimental groups. Cell damage was evaluated by cell proliferation assay (Cell Counting Kit8, CCK8), lactate dehydrogenase (LDH) release assay, 4′, 6diamidino2phenylindole (DAPI) staining, and flow cytometry for apoptosis and reactive oxygen species (ROS) levels. The damage degree of RBC-S on vascular endothelial cells was assessed by statistical analysis of damage data among different groups. Since the damage effect reached a plateau at all time points, the 28 d storage group was selected as the representative for further mechanistic studies. Transcriptomic analysis was performed to explore the role of frizzled class receptor 1 (FZD1) and Wnt signaling pathway in red blood cell storagerelated endothelial dysfunction. Results: Compared with the control group, the storage groups treated with 7 d, 14 d, and 28 d RBC-S showed significantly decreased cell proliferation rates [control group 100%, 7 d group (69.51±2.30)%, 14 d group (74.54±2.89)%, 28 d group (73.59±2.36)%, P<0.05], significantly reduced numbers of DAPI-stained cell nuclei [control group (213±12.5) per field, 7 d group (140.33±17.04) per field, 14 d group (152.00±23.72) per field, 28 d group (144.33±19.09) per field, P<0.05] and significantly increased LDH release [control group (1), 7 d group (8.33±1.41), 14 d group (9.23±0.83), 28 d group (9.16±0.60), P<0.05]. There was no significant difference in the degree of damage caused by RBC-S among different storage groups (P>0.05). With the prolongation of storage time, free hemoglobin (FHb) gradually increased [control group (not detected), 7 d (16.57±6.38) mg/L, 14 d (76.80±22.83) mg/L, 28 d (286.97±29.02) mg/L, P<0.05]. The apoptotic rate (20.53±2.94)% and ROS relative intensity (5.13±0.91) in the 28 d storage group were significantly higher than those in the control group (P<0.05). Transcriptomic analysis showed that FZD1 played a key role in vascular endothelial dysfunction induced by red blood cell storage and was closely related to the Wnt signaling regulatory network. Conclusion: RBC-S stored for 7 d, 14 d, or 28 d can all significantly damage vascular endothelial cells, and the damaging effect reaches a plateau at 7 d of storage. Mechanistic investigation of the 28 d group indicated that the downregulation of the FZD1/Wnt signaling pathway may play a critical role in vascular endothelial dysfunction induced by red blood cell storage, providing a theoretical basis for further optimizing red blood cell storage and transfusion strategies.

Acute respiratory distress syndrome (ARDS) is a common respiratory emergency, but current clinical treatment remains at the level of symptomatic support and there is a lack of effective targeted treatment measures. Our previous study confirmed that inhalation of hydrogen gas can reduce the acute lung injury of ARDS, but the application of hydrogen has flammable and explosive safety concerns. Drinking hydrogen-rich liquid or inhaling hydrogen gas has been shown to play an important role in scavenging reactive oxygen species and maintaining mitochondrial quality control balance, thus improving ARDS in patients and animal models. Coral calcium hydrogenation (CCH) is a new solid molecular hydrogen carrier prepared from coral calcium (CC). Whether and how CCH affects acute lung injury in ARDS remains unstudied. In this study, we observed the therapeutic effect of CCH on lipopolysaccharide (LPS) induced acute lung injury in ARDS mice. The survival rate of mice treated with CCH and hydrogen inhalation was found to be comparable, demonstrating a significant improvement compared to the untreated ARDS model group. CCH treatment significantly reduced pulmonary hemorrhage and edema, and improved pulmonary function and local microcirculation in ARDS mice. CCH promoted mitochondrial peripheral division in the early course of ARDS by activating mitochondrial thioredoxin 2 (Trx2), improved lung mitochondrial dysfunction induced by LPS, and reduced oxidative stress damage. The results indicate that CCH is a highly efficient hydrogen-rich agent that can attenuate acute lung injury of ARDS by improving the mitochondrial function through Trx2 activation.

Acute respiratory distress syndrome(ARDS)is a common respiratory emergency,but current clinical treatment remains at the level of symptomatic support and there is a lack of effective targeted treatment measures.Our previous study confirmed that inhalation of hydrogen gas can reduce the acute lung injury of ARDS,but the application of hydrogen has flammable and explosive safety concerns.Drinking hydrogen-rich liquid or inhaling hydrogen gas has been shown to play an important role in scavenging reactive oxygen species and maintaining mitochondrial quality control balance,thus improving ARDS in patients and animal models.Coral calcium hydrogenation(CCH)is a new solid molecular hydrogen carrier prepared from coral calcium(CC).Whether and how CCH affects acute lung injury in ARDS re-mains unstudied.In this study,we observed the therapeutic effect of CCH on lipopolysaccharide(LPS)induced acute lung injury in ARDS mice.The survival rate of mice treated with CCH and hydrogen inhalation was found to be comparable,demonstrating a significant improvement compared to the untreated ARDS model group.CCH treatment significantly reduced pulmonary hemorrhage and edema,and improved pulmonary function and local microcirculation in ARDS mice.CCH promoted mitochon-drial peripheral division in the early course of ARDS by activating mitochondrial thioredoxin 2(Trx2),improved lung mitochondrial dysfunction induced by LPS,and reduced oxidative stress damage.The results indicate that CCH is a highly efficient hydrogen-rich agent that can attenuate acute lung injury of ARDS by improving the mitochondrial function through Trx2 activation.

Objective To explore the effect of the pneumatic and electric hand training system with multiple sensory stimulation mo-dalities combined with low-frequency repetitive transcranial magnetic stimulation on hand movement and tactile pressure sensation in patients with stroke.Methods From May,2023 to October,2024,62 stroke patients from the First Affiliated Hospital of University of South China were randomly divided into control group(n=31)and experimental group(n=31).Both groups received conventional rehabilitation,and 1 Hz low-frequency repetitive transcranial magnetic stimulation in the primary cerebral motor cortex on the unaffected side,while the experimental group received the pneumatic and electric hand training system with multiple sensory stimulation modalities in addition,for four weeks.They were as-sessed with Fugl-Meyer Assessment-Upper Extremities(FMA-UE),Motor Assessment Scale(MAS),Semmes Weinstein Monofilament Test(SWMT)and modified Barthel index(MBI)before and after treatment.Results One case dropped out in control group and one in the experimental group.After treatment,the scores of FMA-UE,MAS,SWMT and MBI increased in both groups(|t|>5.464,P<0.001),expect the score of SWMT in the control group(P>0.05),and all scores were better in the experimental group than in the control group(|t|>2.195,P<0.05).Conclusion The pneumatic and electric hand training system with multiple sensory stimulation modalities combined with low-frequency repetitive transcranial magnetic stimulation can improve the motor and tactile pressure func-tion of stroke patients,and improve their activity of daily living.

Objective To explore the expression of serine/threonine protein kinase 1(AKT1)and Isocho-rismatase Domain-Containing 1(ISOC1)in gastric cancer tissues and evaluate their diagnostic utility.Methods A total of 90 patients with gastric cancer who were diagnosed and treated in our hospital between March 2023 and September 2024 were recruited as the study subjects.The positive expression rates and positive scores of AKT1 and ISOC1 in cancer tissues and adjacent tissues were compared.The AKT1 and ISOC1 positive scores of gastric cancer patients with diverse characteristics were also compared.The Spearman correlation analysis was employed to exam-ine the relationship between the expression of AKT1 and ISOC1 in cancer tissues and the clinical features of gastric cancer patients.A diagnostic model for gastric cancer,integrating AKT1 and ISOC1,was established using the Logistic regression model.Receiver operating characteristic(ROC)curves were plotted to analyze the area under the curve(AUC)value,sensitivity,and specificity of AKT1,ISOC1,and their combined diagnosis for gastric cancer.Results The positive expression rates and positive scores of AKT1 and ISOC1 in cancer tissues were significantly higher than those in paracancerous tissues(P<0.05).In gastric cancer patients,the expressions of AKT1 and ISOC1 were notably higher in patients with low differentiation,clinical stage Ⅰ～Ⅱ,invasion depth T1-2,absence of lymph node metastasis,and no distant metastasis compared to those with moderate differentiation,clinical stage Ⅲ～Ⅳ,invasion depth T3-4,and presence of lymph node metastasis and distant metastasis(P<0.05).Spearman's correlation analysis indicated that AKT1 and ISOC1 were positively correlated with the degree of differ-entiation,clinical stage,lymph node metastasis,invasion degree,and distant metastasis(P<0.05).The area under the curve(AUC)values of AKT1,ISOC1,and their combined diagnosis for gastric cancer were 0.735,0.726,and 0.875 respectively(P<0.05).The sensitivities were 60.00%,56.70%,and 75.60%,while the speci-ficities were 85.60%,83.30%,and 87.80%.The AUC value of the combined detection of AKT1 and ISOC1 in the diagnosis of gastric cancer was higher than that of AKT1 or ISOC1 alone(Z=-3.003,-3.196,P<0.05).Conclusions AKT1 and ISOC1 are highly expressed in gastric cancer tissues,and their expressions are upregu-lated with the progression of the disease.The combined detection of their expression levels holds great significance for the diagnosis and prognosis assessment of gastric cancer.

Objective To investigate the status of hearing impaired children's hearing device independence skills,and to explore the ways to improve their self-use of hearing equipment.Methods This study surveyed 64 re-habilitation teachers and 411 parents of children with hearing impairment aged 0-12 years.Through face-to-face or remote telephone interview,3 good habits(A asking parents for advice before removing the HA,B putting the de-vice into a moisture-proof box after removing it,C bringing batteries to school and knowing where are them)and 3 key abilities[D wearing the device independently,E replacing the battery independently,and F independently handle foreign bodies in the ear mold(Fa)and water vapor(Fb)]was investigated.The age when mastering skills or de-veloping habits difference of hearing impaired children in different groups were compared.Results ① The ratio of ability D in the bilateral CI group and the bilateral HA group of preschool children was 30.97％and 18.57％respec-tively.Among elementary school children,85.29％and 90.70％had this ability respectively.② The ratio of ability E in the bilateral CI group,the bilateral HA group and the bimodel group were 11.50％,15.71％and 16.49％,re-spectively.Among elementary school children,64.71％,53.49％and 68.52％had this ability,respectively.③Among preschool children,there was no statistical difference in age when different equipment groups developed the three good habits and acquired ability D and E(P＞0.05).④ Among primary school children,there was a statisti-cal difference in the age when different equipment groups formed habit A(P＜0.05),and the age when double CI group had this ability was slightly earlier than the double HA group.There was no significant difference in other abilities among age groups(P＞0.05).Conclusion The age at which hearing impaired children develop the three good habits precedes the age at which they master the key skills,which accords with the law of skill acquisition and development of ordinary children.Corresponding teaching process should be based on the age and ability of hearing-impaired children without considering the type of equipment.

Objective To investigate the status of hearing impaired children's hearing device independence skills,and to explore the ways to improve their self-use of hearing equipment.Methods This study surveyed 64 re-habilitation teachers and 411 parents of children with hearing impairment aged 0-12 years.Through face-to-face or remote telephone interview,3 good habits(A asking parents for advice before removing the HA,B putting the de-vice into a moisture-proof box after removing it,C bringing batteries to school and knowing where are them)and 3 key abilities[D wearing the device independently,E replacing the battery independently,and F independently handle foreign bodies in the ear mold(Fa)and water vapor(Fb)]was investigated.The age when mastering skills or de-veloping habits difference of hearing impaired children in different groups were compared.Results ① The ratio of ability D in the bilateral CI group and the bilateral HA group of preschool children was 30.97％and 18.57％respec-tively.Among elementary school children,85.29％and 90.70％had this ability respectively.② The ratio of ability E in the bilateral CI group,the bilateral HA group and the bimodel group were 11.50％,15.71％and 16.49％,re-spectively.Among elementary school children,64.71％,53.49％and 68.52％had this ability,respectively.③Among preschool children,there was no statistical difference in age when different equipment groups developed the three good habits and acquired ability D and E(P＞0.05).④ Among primary school children,there was a statisti-cal difference in the age when different equipment groups formed habit A(P＜0.05),and the age when double CI group had this ability was slightly earlier than the double HA group.There was no significant difference in other abilities among age groups(P＞0.05).Conclusion The age at which hearing impaired children develop the three good habits precedes the age at which they master the key skills,which accords with the law of skill acquisition and development of ordinary children.Corresponding teaching process should be based on the age and ability of hearing-impaired children without considering the type of equipment.

Objective To explore the expression of serine/threonine protein kinase 1(AKT1)and Isocho-rismatase Domain-Containing 1(ISOC1)in gastric cancer tissues and evaluate their diagnostic utility.Methods A total of 90 patients with gastric cancer who were diagnosed and treated in our hospital between March 2023 and September 2024 were recruited as the study subjects.The positive expression rates and positive scores of AKT1 and ISOC1 in cancer tissues and adjacent tissues were compared.The AKT1 and ISOC1 positive scores of gastric cancer patients with diverse characteristics were also compared.The Spearman correlation analysis was employed to exam-ine the relationship between the expression of AKT1 and ISOC1 in cancer tissues and the clinical features of gastric cancer patients.A diagnostic model for gastric cancer,integrating AKT1 and ISOC1,was established using the Logistic regression model.Receiver operating characteristic(ROC)curves were plotted to analyze the area under the curve(AUC)value,sensitivity,and specificity of AKT1,ISOC1,and their combined diagnosis for gastric cancer.Results The positive expression rates and positive scores of AKT1 and ISOC1 in cancer tissues were significantly higher than those in paracancerous tissues(P<0.05).In gastric cancer patients,the expressions of AKT1 and ISOC1 were notably higher in patients with low differentiation,clinical stage Ⅰ～Ⅱ,invasion depth T1-2,absence of lymph node metastasis,and no distant metastasis compared to those with moderate differentiation,clinical stage Ⅲ～Ⅳ,invasion depth T3-4,and presence of lymph node metastasis and distant metastasis(P<0.05).Spearman's correlation analysis indicated that AKT1 and ISOC1 were positively correlated with the degree of differ-entiation,clinical stage,lymph node metastasis,invasion degree,and distant metastasis(P<0.05).The area under the curve(AUC)values of AKT1,ISOC1,and their combined diagnosis for gastric cancer were 0.735,0.726,and 0.875 respectively(P<0.05).The sensitivities were 60.00%,56.70%,and 75.60%,while the speci-ficities were 85.60%,83.30%,and 87.80%.The AUC value of the combined detection of AKT1 and ISOC1 in the diagnosis of gastric cancer was higher than that of AKT1 or ISOC1 alone(Z=-3.003,-3.196,P<0.05).Conclusions AKT1 and ISOC1 are highly expressed in gastric cancer tissues,and their expressions are upregu-lated with the progression of the disease.The combined detection of their expression levels holds great significance for the diagnosis and prognosis assessment of gastric cancer.

Objective To explore the effect of the pneumatic and electric hand training system with multiple sensory stimulation mo-dalities combined with low-frequency repetitive transcranial magnetic stimulation on hand movement and tactile pressure sensation in patients with stroke.Methods From May,2023 to October,2024,62 stroke patients from the First Affiliated Hospital of University of South China were randomly divided into control group(n=31)and experimental group(n=31).Both groups received conventional rehabilitation,and 1 Hz low-frequency repetitive transcranial magnetic stimulation in the primary cerebral motor cortex on the unaffected side,while the experimental group received the pneumatic and electric hand training system with multiple sensory stimulation modalities in addition,for four weeks.They were as-sessed with Fugl-Meyer Assessment-Upper Extremities(FMA-UE),Motor Assessment Scale(MAS),Semmes Weinstein Monofilament Test(SWMT)and modified Barthel index(MBI)before and after treatment.Results One case dropped out in control group and one in the experimental group.After treatment,the scores of FMA-UE,MAS,SWMT and MBI increased in both groups(|t|>5.464,P<0.001),expect the score of SWMT in the control group(P>0.05),and all scores were better in the experimental group than in the control group(|t|>2.195,P<0.05).Conclusion The pneumatic and electric hand training system with multiple sensory stimulation modalities combined with low-frequency repetitive transcranial magnetic stimulation can improve the motor and tactile pressure func-tion of stroke patients,and improve their activity of daily living.