Objective To construct a Ca2+/calmodulin-dependent protein kinase Ⅱ(CaMKⅡ)long-fragment fusion protein plasmid;investigate the expression,extraction,and purification of CaMK Ⅱ;and identify its binding to the Cav1.2 channel.Methods The extracted pGEX-6p-1/CaMK Ⅱ long-fragment plasmid was transformed into Escherichia coli BL21 receptor cells and cultured in a shaking incubator for 12 h.Isopropyl β-D-thiogalactoside was added to promote GST fusion protein expression.Next,the GST-CaMK Ⅱ long frag-ment was isolated and purified with GS-4B using dithiothreitol(DTT)combined with ultrasonic crushing.After treatment with the PreScis-sion protease,the GST label was removed to obtain the CaMK Ⅱ long-fragment protein.The molecular weight and relative purity of the CaMKⅡ long-fragment protein were determined using 15％sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).The concentration of the purified protein was determined using the Bradford method.The binding ability of the CaMK Ⅱ long-fragment pro-tein to the Cav1.2 channel protein was evaluated using the pull-down method combined with Western blotting.Results The sequencing results showed that the CaMK Ⅱ long fragment was successfully constructed.A CaMK Ⅱ long-fragment protein with high purity and con-centration was obtained using DTT combined with ultrasonic crushing.This protein can bind to the CT1 protein of cardiac Cav1.2 calcium channel.Conclusion In this study,we successfully constructed a CaMKⅡ long-fragment plasmid.The CaMKⅡ long-fragment protein was extracted and purified,and was determined to bind to Cav1.2 channel proteins and exhibit biological activity.Collectively,this study provides a basis for further study of the function of CaMK Ⅱ.

Objective:To evaluate the application value of artificial intelligence (AI) and robot of stereotactic surgery (ROSS)-assisted ultra-early thalamic hematoma drainage in elderly patients with minor thalamus and internal capsule hematoma (TICH) and severe hypoperfusion in the internal capsule area.Methods:A retrospective cross-sectional study was performed; 278 patients with TICH and severe hypoperfusion in the internal capsule area identified by AI medical imaging diagnostic system were enrolled from Department of Neurosurgery, Xingtai Central Hospital from January 2023 to August 2024. Among them, 134 patients (study group) received AI and ROSS-assisted ultra-early thalamic hematoma drainage, and 144 patients (control group) received drug treatment. Differences in baseline data and therapy efficiency between the two groups of patients were compared. Multivariate Logistic regression analysis was used to screen the independent influencing factors for prognosis in the study group 180 days after onset (good prognosis: modified Rankin scale scores of 0-3).Results:Compared with the control group, the study group had significantly higher Glasgow coma scale (GCS) score and regional cerebral blood flow (rCBF) in the internal capsule area 14 days after onset, and statistically higher therapy efficiency and good prognosis rate 180 days after onset ( P<0.05). In the study group, 84 patients had a good prognosis and 50 had a poor prognosis 180 days after onset; compared with the good prognosis group, the poor prognosis group had significantly higher proportion of patients with hyper-homocysteinemia history, lower GCS score on admission, larger volume of thalamic hematoma, higher proportions of patients with combined intraventricular hemorrhage and shunt-dependent hydrocephalus, and lower rCBF in the internal capsule area on admission ( P<0.05). Multivariate Logistic regression analysis showed that thalamic hematoma volume ( OR=2.527, 95% CI: 1.504-4.247, P<0.001), combined intraventricular hemorrhage ( OR=2.325, 95% CI: 1.460-3.703, P<0.001), shunt dependent hydrocephalus ( OR=2.371, 95% CI: 1.267-4.078, P=0.006), and rCBF in the internal capsule area on admission ( OR=0.672, 95% CI: 0.314-1.025, P<0.001) were independent risk factors for prognosis of patients in the study group. Conclusion:AI and ROSS-assisted ultra-early thalamic hematoma drainage is effective for elderly patients with minor TICH and severe hypoperfusion in the internal capsule area; patients with large thalamic hematoma volume, combined intraventricular hemorrhage, shunt-dependent hydrocephalus, and low rCBF in the internal capsule area on admission are prone to have a poor prognosis.

Abstract To understand the relationship between cardiovascular disease (CVD) and adverse childhood experiences (ACEs), the present review aims to describe the burden and influencing factors of CVD, epidemiological characteristics and burden of ACEs, current research on the relationship between ACEs and CVD, and the mechanism of ACEs leading to CVD. It is proposed that further assessment of the relationship is warranted through identifying blood biomarkers, conducting prospective cohort studies and intervention studies. Such efforts would provide valuable scientific insights for primary prevention strategies for cardiovascular disease.

Objective To evaluate the clinical value of bedside ultrasonographic measurement of optic nerve sheath diameter(ONSD)combined with serum matrix metalloproteinase-9(MMP-9)and neutrophil-to-lymphocyte ratio(NLR)in assessing postoperative intracranial hypertension in patients with severe traumatic brain injury.Methods A total of 100 patients with severe traumatic brain injury were enrolled as study subjects.According to postoperative intracranial pressure status,the patients were divided into normal group(n=33)and elevated group(n=67).ONSD,MMP-9 and NLR lev-els were compared between the two groups.The correlations among ONSD,serum MMP-9 and NLR in the elevated group were analyzed.The influencing factors of postoperative intracranial hypertension in patients with severe traumatic brain injury were evaluated.Receiver operating characteristic(ROC)curve analysis was performed to assess the diagnostic value of ONSD combined with serum MMP-9 and NLR for postoperative intracranial hypertension.Results ONSD,serum MMP-9 and NLR levels were significantly higher in the elevated group than those in the normal group(P＜0.05).ONSD was positively correlated with MMP-9(r=0.367,P=0.002),ONSD was positively correlated with NLR(r=0.419,P＜0.01),and MMP-9 was positively correlated with NLR(r=0.324,P=0.007).Elevated ONSD,serum MMP-9 and NLR were risk factors for increased in-tracranial pressure after severe craniocerebral injury surgery(P＜0.05).The area under the curve for evaluation of postoperative intracranial pressure increase in patients with severe craniocerebral in-jury by combination of ONSD,serum MMP-9 and NLR was 0.976(95％CI,0.952 to 1.000),the sensitivity was 91.04％,and the specificity was 93.94％.The combined evaluation value of ONSD,serum MMP-9 and NLR for increased intracranial pressure after surgery in patients with severe craniocerebral injury was higher(Zcombined with-ONSD=3.453,P=0.001,Zcombined with-MMP-9=3.637,P＜0.001,Zcombined with-NLR=2.654,P=0.008).Conclusion In patients with increased intracra-nial pressure after severe craniocerebral injury surgery,their levels of ONSD,serum MMP-9 and NLR increase,and the increase of the three indicators will increase the risk of increased intracranial pressure after surgery.The combined detection of the three indicators has certain value in evaluating the increase of intracranial pressure in patients after surgery.

Dental treatments for children and adolescents have unique clinical characteristics that differ from dental care for adults in terms of children's physiology, psychology, and behavior. These differences impose specific requirements on the application of local anesthesia in pediatric dental procedures. This article presents expert consensus on the principles of local anesthesia techniques in pediatric dental therapies, including the use of common anesthetic drugs and dosage control, safety and efficacy evaluation, and prevention and management of complications. The aim is to improve the safety and quality of pediatric dental treatments and offer guidance for clinical application by dentists.
Humans ; Child ; Anesthesia, Local/methods* ; Consensus ; Anesthesia, Dental/methods* ; Adolescent ; Anesthetics, Local/administration & dosage* ; Dental Care for Children

This study establishes a high-performance liquid chromatography (HPLC) method for the determination of acetate content in hemodialysis solutions and dialysis concentrates. In this study, Synergi Polar-RP column is utilized. Phosphate buffered saline (50 mmol/L, pH=2.5) is used as a mobile phase. The flow rate is 1.0 mL/min. The wavelength of detection is 212 nm. Results show that the linear relationship of acetate is good in the range of 0.1~20 mmol/L, r =0.999 9 and the spike recoveries are from 98.9%~99.5%, RSD<0.5% ( n=3). This method can easily and accurately determine the acetate content in hemodialysis solutions and dialysis concentrates, and can be applied to quality control in the production and use of such products.
Chromatography, High Pressure Liquid/methods* ; Acetates/analysis* ; Hemodialysis Solutions/analysis* ; Dialysis Solutions/analysis* ; Renal Dialysis

Objective:To investigate the clinical value of a fully autonomous ultrasound robot in splenic ultrasound imaging.Methods:A retrospective study was conducted by enrolling 56 adult volunteers from the Third Medical Center of the Chinese PLA General Hospital between February 1-8，2024 as research subjects.A senior physician sequentially performed splenic ultrasound examinations using both the fully autonomous ultrasound robot and a matched portable ultrasound device. The acquired images were randomly coded and scored via a double-blind method by 3 physicians. The differences of the image quality scores and high-quality image proportions between the two groups were compared. Examination durations were recorded and compared between the two groups.Results:Both modalities successfully acquired splenic images in all 56 volunteers. No statistically significant differences were observed in image quality scores among the 3 physicians：（3.52 ± 1.31）points vs.（3.83 ± 1.23）points，（2.77 ± 1.23）points vs.（3.17 ± 1.17）points，and（3.48 ± 0.97）points vs.（3.79 ± 0.94）points（all P>0.05）. The numbers of images scoring ≥ 3 points showed no significant differences：45（80.36%） vs. 50（89.29%），30（53.57%） vs. 38（67.86%），and 48（85.71%） vs. 52（92.86%）（all P>0.05）. The fully autonomous ultrasound robot required significantly longer examination time［（60.86 ± 50.55）s vs.（7.95 ± 4.35）s， t=6.88， P＜0.01］. Conclusions:The fully autonomous ultrasound robot demonstrates comparable image quality and clinically acceptable image proportions to conventional portable ultrasound in splenic examinations. These findings suggest its potential equivalence to operator-dependent ultrasound for splenic imaging，supporting its feasibility as an alternative ultrasound modality despite longer procedural duration.

Objective:To explore the clinical characteristics and prognosis of primary renal dedifferentiated liposarcoma.Methods:A retrospective analysis was conducted on the clinical data of 10 patients diagnosed with renal dedifferentiated liposarcoma in the First Affiliated Hospital of Zhengzhou University from January 2017 to December 2023. The cohort consisted of 8 males and 2 females, with a mean age of (59.0±6.8) years. Tumors were located in the left kidney in 8 cases and in the right kidney in 2 cases. Presentations included flank or abdominal masses in 4 patients, back pain in 3 patients, and asymptomatic in 3 patients. Imaging studies revealed solitary lesions in 9 cases and multiple lesions in 1 case. The maximum tumor diameter ranged from 95 to 178 mm, with a median of 119.5 mm. CT showed tumors within the renal parenchyma with unclear boundaries and displayed a "slow in, slow out" pattern of gradual enhancement. Clinical staging revealed T 2N 0M 0 in 3 cases, T 3N 0M 0 in 4 cases, T 4N 0M 0 in 3 cases, and T 2N 1M 0 in 1 case. Nine patients underwent radical nephrectomy, including 4 laparoscopic surgeries, 4 open surgeries, and 1 robotic-assisted with thrombectomy. One patient, presenting with multiple lymph node metastases confirmed by PET-CT, underwent a diagnostic biopsy. Postoperative pathological features, follow-up and prognosis were analyzed. Results:Pathological specimens appeared grayish-red, grayish-yellow or grayish-white cut surfaces with soft to moderate texture. Microscopically, tumor cells were ovoid or short spindle-shaped with significant atypia and cytoplasmic vacuoles, with visible pathological mitoses. Six cases showed only dedifferentiated components with tumor giant cells and multinucleated giant cells resembling pleomorphic undifferentiated sarcoma, with focal tumor necrosis. Fluorescence in situ hybridization showed MDM2 gene amplification in all cases. All cases were diagnosed as primary renal dedifferentiated liposarcoma. Pathological staging showed 4 cases as pT 2N 0M 0, 3 cases as pT 3N 0M 0, 2 cases as pT 4N 0M 0, and 1 case lacked pathological staging due to biopsy only. Five patients received postoperative adjuvant therapy, including two pT 2N 0M 0 cases who received immunotherapy and apatinib treatment respectively, with no recurrence. One pT 3N 0M 0 case received anlotinib treatment, with local recurrence after 12 months, followed by radiofrequency ablation combined with chemotherapy. Two pT 4N 0M 0 cases received ifosfamide combined with epirubicin and pirarubicin combined with lobaplatin respectively, with one case showing no progression at 11 months follow-up, and another case developing splenic metastasis 3 months post-surgery, followed by chemotherapy combined with targeted therapy, surviving for 20 months. Among the 4 cases without adjuvant therapy, two pT 2N 0M 0 cases developed multiple metastases within 1 month post-surgery and received immunotherapy combined with targeted therapy and/or chemotherapy, surviving 4-5 months.One of the two pT 3N 0M 0 cases developed local recurrence 2 months post-surgery and received chemotherapy, surviving 6 months, and another pT 3N 0M 0 case developed gluteal subcutaneous metastasis 1 month post-surgery and received immunotherapy combined with targeted therapy, surviving 8 months.One non-surgical pT 2N 1M 0 patient received chemotherapy and survived for 15 months. All patients were followed up for 4-52 months, with a median follow-up time of 11 months. At the last follow-up, 6 patients died and 4 survived. Conclusions:Primary renal dedifferentiated liposarcoma is clinically rare, with atypical symptoms and difficult preoperative diagnosis, relying on pathology for confirmation. Radical nephrectomy is the main treatment method, but surgery alone has poor prognosis with high recurrence and metastasis rates. Adjuvant therapy based on surgery may improve patient prognosis.Larger sample studies are needed for confirmation.

Objective:To reveal the coupling mechanism of beam temporal profile and tissue oxygen content on radical kinetics, further explain the potential biological basis of the FLASH effect, and provide a reference for beam optimization and treatment planning design of FLASH radiotherapy (FLASH-RT).Methods:TOPAS-nBio v3.0 was used to simulate the physical and chemical processes of electron beams in water, and a full-scale kinetic model was established covering the generation, diffusion, reaction, and quenching of free radicals such as hydroxyl radical (·OH) and hydrated electrons (e aq-). Under different beam temporal profiles (single pulse, multi-pulses, continuous wave irradiation) and different oxygen concentration conditions, the evolution dynamics of free radicals were systematically simulated. At the same time, the data on e aq- content were obtained by experimental measurement of laser absorption spectroscopy to verify the accuracy of the model prediction. Results:The changing trend of e aq- concentration measured in the experiment was highly consistent with the simulation result, verifying the reliability of the constructed model. The beam time structure had a significant impact on the peak value and duration of free radical concentration. The single-pulse structure can cause the free radicals to rapidly increase and then quickly quench in a short time, while the continuous or long-pulse structure can cause the radical concentration to remain at a high level for a long time. The evolution of ·OH was not sensitive to the oxygen environment, while e aq- are greatly affected by the oxygen environment. The scavenging efficiency of free radicals in a hypoxic environment was significantly decreased, leading to an enhanced accumulation of oxidative damage to biological macromolecules. The lifespan of e aq- in an oxygen-rich environment decreased rapidly. Conclusions:Radical kinetics are regulated by both the beam temporal profile and oxygen content. FLASH-RT can utilize single-pulse or multi-pulses intervals to form periodic windows, reducing normal tissue damage by efficiently scavenging free radicals through antioxidants, while free radicals in tumor tissues continuously accumulate and amplify damage, thus generating a selective protective effect.