ObjectiveTo investigate the effects of targeted silencing of Runt-related Transcription Factor 3 （RUNX3） on the proliferation and migration of Mouse Hepatic Stellate Cells （HSCs）， as well as subsequent collagen deposition. MethodsMouse hepatic stellate cell line （JS-1） was selected and then morphologically observed and identified under a microscope. After the cells had fully adhered， they were treated with 5 ng/mL of transforming growth factor beta 1 （TGF-β₁） for 24 hours to induce hepatic stellate cell activation. Furthermore， a RUNX3 silencing model was established using RUNX3 lentiviral infection. The experiment was divided into four groups： Control group， TGF-β₁ group， TGF-β₁+siRNA-NC group， and TGF-β₁+siRNA-RUNX3 group. Protein expression changes of RUNX3， alpha-smooth muscle actin （α-SMA）， and Alpha 1 type I collagen （Collagen I） were detected using Western blot method. Cellular immunofluorescence assays were employed to investigate the deposition changes of α-SMA and RUNX3 in hepatic stellate cells. RT-qPCR was utilized to examine the mRNA expression changes of RUNX3， α-SMA， and Collagen I. The proliferative capacity of hepatic stellate cells was assessed using Edu staining. The migratory ability of hepatic stellate cells was evaluated through wound healing assays and Transwell migration experiments. ResultsCompared with Control group， a significant elevation in RUNX3 was observed in the TGF-β₁-induced activated HSCs （P<0.01）. Meanwhile， the protein and mRNA levels of fibrosis-related markers and α-SMA and Collagen I were significantly upregulated （P<0.001）. Additionally， the proliferation and migration capabilities of HSCs were significantly enhanced （P<0.001）. In contrast， when compared to TGF-β₁+siRNA-NC group， TGF-β₁+siRNA-RUNX3 group exhibited a notable decrease in RUNX3 and other related indicators， such as the protein and mRNA levels of α-SMA and Collagen I （P<0.05）. Concurrently， the proliferation and migration capabilities of HSCs were significantly inhibited in TGF-β₁+siRNA-RUNX3 group （P<0.01）. ConclusionSilencing RUNX3 can inhibit the deposition of collagen and the proliferation and migration of hepatic stellate cells. Conversely， RUNX3 promotes the proliferation and migration capabilities of HSCs， thereby facilitating the activation of HSC.

This study explored the drying kinetics of Salviae Miltiorrhizae Radix et Rhizoma(SM), established the suitable models simulating the drying kinetics, and then analyzed the dynamic changes of active components during the drying processes with different methods, aiming to provide a basis for the establishment of suitable drying methods and the quality control of SM. The drying kinetics were studied based on the drying curve, drying rate, moisture effective diffusion coefficient, and drying activation energy, and the appropriate drying kinetics model of SM was established. The drying performance of different methods, such as hot air drying, infrared drying, and microwave drying of SM was evaluated, and the changes in the content of 10 salvianolic acids and 6 tanshinones during drying were analyzed by UPLC-TQ-MS. The Technique for Order Preference by Similarity to an Ideal Solution(TOPSIS) was employed to evaluate the quality of SM dried with different methods. The results showed that the drying rate and moisture effective diffusion coefficient of SM increased with the rise in drying temperature, and the maximum drying rates of different methods were in the order of microwave drying > infrared drying > hot air drying, slice > whole root. The drying rate decreased with the rise in temperature and the extension of drying time. The activation energy of hot air drying was higher than that of infrared drying in SM. The most suitable model for simulating the drying process of SM was the Page model. The TOPSIS results suggested infrared drying at 50 ℃ was the optimal drying method for SM. During the drying process, the content of salvianolic acids increased in different degrees with the loss of moisture, among which salvianolic acid B showed the largest increase of 44 times compared with that in the fresh medicinal material. Tanshinones also existed in the fresh herb of SM, and the content of tanshinone Ⅱ_A increased by 3 times after drying. The results provided a basis for the establishment of suitable drying methods and the quality control of SM.
Salvia miltiorrhiza/chemistry* ; Desiccation/methods* ; Drugs, Chinese Herbal/chemistry* ; Rhizome/chemistry* ; Kinetics ; Quality Control ; Abietanes

Objective To observe the effect of RAW264.7 cells on calcium sparks in a insulin resistance model of C2C12 cells induced by sodium palmitate.Methods C2C12 cells and RAW264.7 cells were co-cultured to simulate the in vivo state of skeletal muscle.C2C12 cells were cultured in high-glucose medium containing 2%horse serum to induce differentiation into mature myotubes,and then divided into 5 groups:control(RAW264.7 cells),co-culture of C2C12 with RAW264.7,C2C12 alone,co-culture of C2C12 with RAW264.7 plus sodium palmitate(PA),and C2C12 alone with PA.PA of 5 mmol/L was used to induce insulin resistance in C2C12 cells for 24 hours.Revived and expanded RAW264.7 cells were evenly added to C2C12 cells and co-cultured for two days.Subsequently,cells were maintained in modified suspension culture,and both cell types were loaded with the calcium ion fluorescent probe Fluo-4 AM.Finally,Paraxanthine was used to induce intracellular calcium sparks,which was captured and recorded under a laser confocal microscope.Results No significant calcium signal change was observed in the control group.Co-cultured C2C12 cells exhibited rapid and pronounced calcium signal changes,whereas calcium signals in C2C12 cells cultured alone increased slowly throughout the observation period without a sharp decline.The peak calcium signal was reached significantly faster in co-cultured C2C12 cells than that in C2C12 cells cultured alone(P<0.001).With PA induction,calcium signal changes in C2C12 cells were not markedly altered,while distinct calcium fluctuations were still observed in co-cultured C2C12 cells,and the peak calcium signal was reached significantly faster in co-cultured C2C12 cells than that in C2C12 cells cultured alone(P<0.001).Conclusion RAW264.7 cells enhance the dynamic responsiveness of calcium signaling in both normal and PA-stimulated C2C12 cells.

Objective To observe the effect of RAW264.7 cells on calcium sparks in a insulin resistance model of C2C12 cells induced by sodium palmitate.Methods C2C12 cells and RAW264.7 cells were co-cultured to simulate the in vivo state of skeletal muscle.C2C12 cells were cultured in high-glucose medium containing 2%horse serum to induce differentiation into mature myotubes,and then divided into 5 groups:control(RAW264.7 cells),co-culture of C2C12 with RAW264.7,C2C12 alone,co-culture of C2C12 with RAW264.7 plus sodium palmitate(PA),and C2C12 alone with PA.PA of 5 mmol/L was used to induce insulin resistance in C2C12 cells for 24 hours.Revived and expanded RAW264.7 cells were evenly added to C2C12 cells and co-cultured for two days.Subsequently,cells were maintained in modified suspension culture,and both cell types were loaded with the calcium ion fluorescent probe Fluo-4 AM.Finally,Paraxanthine was used to induce intracellular calcium sparks,which was captured and recorded under a laser confocal microscope.Results No significant calcium signal change was observed in the control group.Co-cultured C2C12 cells exhibited rapid and pronounced calcium signal changes,whereas calcium signals in C2C12 cells cultured alone increased slowly throughout the observation period without a sharp decline.The peak calcium signal was reached significantly faster in co-cultured C2C12 cells than that in C2C12 cells cultured alone(P<0.001).With PA induction,calcium signal changes in C2C12 cells were not markedly altered,while distinct calcium fluctuations were still observed in co-cultured C2C12 cells,and the peak calcium signal was reached significantly faster in co-cultured C2C12 cells than that in C2C12 cells cultured alone(P<0.001).Conclusion RAW264.7 cells enhance the dynamic responsiveness of calcium signaling in both normal and PA-stimulated C2C12 cells.

Objective To explore the value of analysis on the incidence of healthcare-associated infection(HAI)based on disease diagnosis-related grouping(DRG),case mix index(CMI),and relative weight(RW).Methods All discharged cases,DRG and HAI status in a tertiary first-class general hospital from January 1 to December 31,2023 were analyzed retrospectively.Incidences of HAI in different departments were adjusted and compared by CMI.Incidences of HAI in different DRG groups were adjusted by RW.Results Among the 47 695 cases included in the analysis,757 were HAI cases,including 225 DRG groups.The department of critical care medicine had the highest incidence of HAI(11.98％).After CMI adjustment,departments with higher incidence of HAI were main-ly the department of respiratory and critical care medicine(3.96％),department of critical care medicine(3.04％),and department of neurology(2.85％),et al.DRG groups with the top five high incidence of HAI were AH11(tracheotomy and with ventilator support ≥96 hours or extracorporeal membrane oxygenation[ECMO],accompa-nied by major complications and comorbidity[MCC],50.00％),BC29(ventricular shunt and revision surgery,31.43％),BB21(craniotomy other than trauma,accompanied by MCC,27.56％),BB11(craniotomy of brain trauma,accompanied by MCC,26.32％),and GB1A(major surgery of esophagus,stomach,and duodenum,accompanied by major or moderate complications and comorbidity,16.00％).After RW adjustment,the DRG groups with the top five high incidence of HAI were ES21(respiratory system infection/inflammation,accompanied by MCC,5.89％),BR21(cerebral ischemic disease,accompanied by MCC,5.17％),FR11(heart failure,shock,accompanied by MCC,4.80％),BC29(4.57％)and AH11(3.57％).Conclusion Analyzing the incidence of HAI based on CMI and RW can help to identify key departments and disease groups for infection prevention and control,and provide reference for precise prevention and control of HAI in the new era.

Aim To investigate the effect of N6-methy-ladenosine(m6A)demethylase ALKBH5 on the prolif-eration and migration of cardiac fibroblasts(CFs)in-duced by high glucose.Methods Primary CFs were isolated from neonatal mouse hearts and identified u-sing optical and confocal microscopy.Cell activation was induced using a high-glucose medium(33 mmol·L-1 glucose).An ALKBH5 overexpression model was established by transfecting CFs with an ALKBH5 ex-pression vector in a high-glucose medium.The expres-sion of ALKBH5 in CFs was assessed through immuno-fluorescence staining,Western blot and RT-qPCR.Changes in m6A levels were evaluated using Dot blot a-nalysis.Additionally,Alterations in the expression of proliferating cell nuclear antigen(PCNA)and collagenⅠ,a pivotal fibrosis indicator,were measured using Western blot.The proliferation and migration ability of CFs were assessed through EdU staining and Transwell migration assay,respectively.Results Following treatment with high glucose,the expression of ALKBH5 in CFs notably decreased,while m6A level increased.This was accompanied by a significant increase in the expression of the proliferation marker PCNA and the fi-brosis marker collagen Ⅰ.Additionally,there was a sig-nificant improvement in the ability of proliferation and migration.Overexpression of ALKBH5 resulted in a significant decrease in the expressions of PCNA and collagen Ⅰ,leading to the inhibition of both proliferation and migration in CFs.Conclusion Overexpression of ALKBH5 suppresses the expression of PCNA and colla-gen Ⅰ,consequently reducing the proliferation and mi-gration of CFs,potentially through m6A methylation modification.

Objective To explore the value of analysis on the incidence of healthcare-associated infection(HAI)based on disease diagnosis-related grouping(DRG),case mix index(CMI),and relative weight(RW).Methods All discharged cases,DRG and HAI status in a tertiary first-class general hospital from January 1 to December 31,2023 were analyzed retrospectively.Incidences of HAI in different departments were adjusted and compared by CMI.Incidences of HAI in different DRG groups were adjusted by RW.Results Among the 47 695 cases included in the analysis,757 were HAI cases,including 225 DRG groups.The department of critical care medicine had the highest incidence of HAI(11.98％).After CMI adjustment,departments with higher incidence of HAI were main-ly the department of respiratory and critical care medicine(3.96％),department of critical care medicine(3.04％),and department of neurology(2.85％),et al.DRG groups with the top five high incidence of HAI were AH11(tracheotomy and with ventilator support ≥96 hours or extracorporeal membrane oxygenation[ECMO],accompa-nied by major complications and comorbidity[MCC],50.00％),BC29(ventricular shunt and revision surgery,31.43％),BB21(craniotomy other than trauma,accompanied by MCC,27.56％),BB11(craniotomy of brain trauma,accompanied by MCC,26.32％),and GB1A(major surgery of esophagus,stomach,and duodenum,accompanied by major or moderate complications and comorbidity,16.00％).After RW adjustment,the DRG groups with the top five high incidence of HAI were ES21(respiratory system infection/inflammation,accompanied by MCC,5.89％),BR21(cerebral ischemic disease,accompanied by MCC,5.17％),FR11(heart failure,shock,accompanied by MCC,4.80％),BC29(4.57％)and AH11(3.57％).Conclusion Analyzing the incidence of HAI based on CMI and RW can help to identify key departments and disease groups for infection prevention and control,and provide reference for precise prevention and control of HAI in the new era.

Aim To investigate the effect of N6-methy-ladenosine(m6A)demethylase ALKBH5 on the prolif-eration and migration of cardiac fibroblasts(CFs)in-duced by high glucose.Methods Primary CFs were isolated from neonatal mouse hearts and identified u-sing optical and confocal microscopy.Cell activation was induced using a high-glucose medium(33 mmol·L-1 glucose).An ALKBH5 overexpression model was established by transfecting CFs with an ALKBH5 ex-pression vector in a high-glucose medium.The expres-sion of ALKBH5 in CFs was assessed through immuno-fluorescence staining,Western blot and RT-qPCR.Changes in m6A levels were evaluated using Dot blot a-nalysis.Additionally,Alterations in the expression of proliferating cell nuclear antigen(PCNA)and collagenⅠ,a pivotal fibrosis indicator,were measured using Western blot.The proliferation and migration ability of CFs were assessed through EdU staining and Transwell migration assay,respectively.Results Following treatment with high glucose,the expression of ALKBH5 in CFs notably decreased,while m6A level increased.This was accompanied by a significant increase in the expression of the proliferation marker PCNA and the fi-brosis marker collagen Ⅰ.Additionally,there was a sig-nificant improvement in the ability of proliferation and migration.Overexpression of ALKBH5 resulted in a significant decrease in the expressions of PCNA and collagen Ⅰ,leading to the inhibition of both proliferation and migration in CFs.Conclusion Overexpression of ALKBH5 suppresses the expression of PCNA and colla-gen Ⅰ,consequently reducing the proliferation and mi-gration of CFs,potentially through m6A methylation modification.

Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75％(27/36);boys accounted for 64％(23/36).Among the 36 neonates,31(86％)had disease onset within 3 days after birth,and 19(53％)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61％)had left cerebral infarction and 13(36％)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75％)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72％)and seizures in 10 neonates(34％).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33％,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44％(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.

Objective:To assess the efficacy of modified arthroscopic lower trapezius transfer with autologous hamstring tendon bridging for irreparable massive posterosuperior rotator cuff tears.Methods:A retrospective analysis was conducted on 59 patients (11 males and 48 females; mean age 66.70±6.10 years, range 55-79 years) treated between June 2019 and November 2022 for posterior superior irreparable massive rotator cuff tears. The treatment involved total arthroscopic oblique muscle transposition combined with partial rotator cuff repair. Shoulder mobility was measured in active supination, forward flexion, posterior external rotation, and abduction external rotation before surgery and at the final follow-up. The Constant-Murley shoulder function score, visual analogue scale (VAS) for pain, and American Shoulder and Elbow Surgeons (ASES) scores were assessed preoperatively, at 3 months, 6 months postoperatively, and at the final follow-up. MR examinations were conducted preoperatively and at 3, 6, and 12 months postoperatively to evaluate rotator cuff healing and retear rates.Results:All surgeries were successfully completed using an average of 3.2±0.5 anchor nails (range 3-5) per case. The mean follow-up period was 15.9±5.7 months (range 12-37 months). Significant improvements in shoulder mobility were observed postoperatively compared to preoperative measurements. The mean body external rotation angle increased from 12.0°±19.4° preoperatively to 35.3°±19.6° at the final follow-up ( P<0.05), and the mean abduction external rotation angle increased from 33.4°±22.4° to 43.4°±23.1° ( P<0.05). The mean preoperative abduction angle improved from 121.7°±47.9° to 136.4°±40.6° ( P<0.05), and the mean forward flexion supination angle improved from 117.6°±45.8° to 139.5°±33.7° ( P<0.05). Postoperative VAS scores significantly improved from 4.8±1.8 preoperatively to 0.3±0.8 at the final follow-up ( F=104.868, P<0.001). The ASES scores improved from 42.0±14.9 preoperatively to 71.7±14.6 at the final follow-up ( F=47.287, P<0.001). The Constant-Murley scores increased from 54.3±17.1 preoperatively to 76.4±13.0 at the final follow-up ( F=44.082, P<0.001). Postoperative complications included 2 complete tears and 7 partial tears (re-tear rate 15.3%). No serious complications such as shoulder joint infection, joint stiffness, or vascular nerve injury were observed. One patient developed a localized subcutaneous hematoma at the tendon extraction site, and another developed deep vein thrombosis of the lower extremity, which improved with symptomatic anticoagulation. Conclusion:Modified arthroscopic lower trapezius transfer with autologous hamstring tendon bridging effectively reduces shoulder pain, improves shoulder joint activity and function scores, and is associated with a low rate of surgical complications for patients with irreparable massive posterosuperior rotator cuff tears.