OBJECTIVE To establish a rapid analytical method to identify the chemical components in Xitong preparations for the treatment of rheumatoid arthritis by ultra-high performance liquid chromatography coupled triple time-of-flight mass spectrometry(UPLC-Triple-TOF-MS/MS). METHODS The analysis was performed on an ACQUITY UPLC HSS T3 (2.1 mm×150 mm, 1.8 μm) column with the mobile phase consisting of water containing 0.1% formic acid(A) and acetonitrile containing 0.1% formic acid(B) in gradient mode at a flow rate of 0.3 mL·min−1. Data acquisition was carried out under positive and negative ion modes. The processed data was analyzed by Peakview software and compound database. The chemical components were determined by comparison with standard products, combining with the characteristic fragments in secondary MS, and those in the related literatures. RESULTS Seventy-eight compounds were identified including 20 phenylpropanoids, 33 terpenes, 19 flavonoids, 6 oxylipins, two of which might be new compounds. CONCLUSION The method can be used for rapid identification of chemical compositions in Xitong preparations, which provides basis for quality control and elucidation of phamarcodynamic material basis of Xitong pills and capsules .

Cerebral small vessel disease （CSVD） is a common disease with great impact on the health of the Chinese population. CSVD has insidious progression and is often neglected by both patients and physicians. In recent years， advances have been made in the research on CSVD from the aspects of risk factors， pathogenesis， clinical manifestations， and evaluation systems. In 2013， the international Standards for Reporting Vascular Changes on Neuroimaging （STRIVE） collaborative group standardized the definition and description of CSVD， and subsequently in 2021， our team released Chinese expert consensus on the diagnosis and treatment of cerebral small vessel disease 2021， which summarized the latest research findings in China and globally. In 2023， the international STRIVE collaborative group provided further updates on research advances in the field of CSVD. CSVD has similar clinical manifestations to neurodegenerative diseases， with a lack of significant specificity. Although genetic testing and brain tissue biopsy help to make a confirmed diagnosis to a certain extent， their application in clinical practice has been limited by technical and financial constraints. At present， neuroimaging techniques are mainly used to detect brain tissue lesions induced by CSVD and make a diagnosis. This article discusses the imaging markers for the diagnosis of CSVD and the imaging-based differential diagnosis of CSVD.
Diagnosis

Tripterygium glycosides tablet(TGT),the classical commercial drug of Tripterygium wilfordii Hook.F.has been effectively used in the treatment of rheumatoid arthritis,nephrotic syndrome,leprosy,Behcet's syndrome,leprosy reaction and autoimmune hepatitis.However,due to its narrow and limited treatment window,TGT-induced organ toxicity(among which liver injury accounts for about 40％of clinical reports)has gained increasing attention.The present study aimed to clarify the cellular and molecular events underlying TGT-induced acute liver injury using single-cell RNA sequencing(scRNA-seq)technology.The TGT-induced acute liver injury mouse model was constructed through short-term TGT exposure and further verified by hematoxylin-eosin staining and liver function-related serum indicators,including alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase and total bilirubin.Using the mouse model,we identified 15 specific subtypes of cells in the liver tissue,including endothelial cells,hepatocytes,cholangiocytes,and hepatic stellate cells.Further analysis indicated that TGT caused a significant inflammatory response in liver endothelial cells at different spatial locations;led to marked inflammatory response,apoptosis and fatty acid metabolism dysfunction in hepatocytes;activated he-patic stellate cells;brought about the activation,inflammation,and phagocytosis of liver capsular macrophages cells;resulted in immune dysfunction of liver lymphocytes;disturbed the intercellular crosstalk in liver microenvironment by regulating various signaling pathways.Thus,these findings elaborate the mechanism underlying TGT-induced acute liver injury,provide new insights into the safe and rational applications in the clinic,and complement the identification of new biomarkers and ther-apeutic targets for liver protection.

Objective:To analyze the incidence and epidemiological characteristics of traumatic spinal cord injury in China in 2018.Methods:Multi-stage stratified cluster sampling was used to randomly select hospitals capable of treating patients with spinal cord injury from 3 regions，9 provinces and 27 cities in China to retrospectively investigate eligible patients with traumatic spinal cord injury admitted in 2018. National and regional incidence rates were calculated. The data of cause of injury，injury level，severity of injury，segment and type of fracture，complications，death and other data were collected by medical record questionnaire，and analyzed according to geographical region，age and gender.Results:Medical records of 4，134 patients were included in this study，with a male-to-female ratio of 2.99∶1. The incidence of traumatic spinal cord injury in China in 2018 was 50.484 / 1 million （95% CI 50.122-50.846）. The highest incidence in the Eastern region was 53.791 / 1 million （95% CI 53.217-54.365）. In the whole country，the main causes of injury were high falls （29.58%），as well as in the Western region （40.68%），while the main causes of injury in the Eastern and Central regions were traffic injuries （31.22%，30.10%）. The main injury level was cervical spinal cord in the whole country （64.49%），and the proportion of cervical spinal cord injury in the Central region was the highest （74.68%），and the proportion of lumbosacral spinal cord injury in the Western region was the highest （32.30%）. The highest proportion of degree of injury was incomplete quadriplegia （55.20%），and the distribution pattern was the same in each region. A total of 65.87% of the patients were complicated with fracture or dislocation，77.95% in the Western region and only 54.77% in the Central region. In the whole country，the head was the main combined injury （37.87%），as well as in the Eastern and Central regions，while the proportion of chest combined injury in the Western region was the highest （38.57%）. A total of 32.90% of the patients were complicated with respiratory complications. There were 23 patients （0.56%） died in hospital，of which 17（73.91%） died of respiratory dysfunction. Conclusions:The Eastern region of China has a high incidence of traumatic spinal cord injury. Other epidemiological features include high fall as the main cause of injury cervical spinal cord injury as the main injury level，incomplete quadriplegia as the main degree of injury，head as the main combined injury，and respiratory complications as the main complication.

OBJECTIVE: To investigate the potential risk of approach-related complications at different access angles in minimally invasive lateral lumbar interbody fusion.METHODS: Eighty-six axial magnetic resonance images were obtained to analyze the risk of approach-related complications. The access corridor were simulated at different access angles and the potential risk of neurovascular structure injury was evaluated when the access corridor touching or overlapping the corresponding structures at each angle. Furthermore, the safe corridor length was measured when the corridor width was 18 and 22 mm.RESULTS: When access angle was 0°, the potential risk of ipsilateral nerve roots injury was 54.7% at L4–L5. When access angle was 45°, the potential risk of abdominal aorta, contralateral nerve roots or central canal injury at L4–L5 was 79.1%, 74.4%, and 30.2%, respectively. The length of the 18 mm-wide access corridor was largest at 0° and it could reach 44.5 mm at L3–L4 and 46.4 mm at L4–L5. While the length of the 22 mm-wide access corridor was 42.3 mm at L3–L4 and 44.1 mm at L4–L5 at 0°.CONCLUSION: Changes in the access angle would not only affect the ipsilateral neurovascular structures, but also might adversely influence the contralateral neural elements. It should be also noted to surgeons that alteration of the access angle changed the corridor length.
Aorta, Abdominal ; Magnetic Resonance Imaging ; Surgeons

Aim To investigate the role of COX-2 in BMP9 induced osteogenic differentiation,and the pos-sible mechanism underlying this function of COX-2. Methods We introduced real-time PCR, Western blot, and immunocytochemical staing to detect the effect of BMP9 on COX-2 expression.We employed chemiluminescence technique to assay ALP activities, RT-PCR to detect the expression of Smad6 and Smad7 , and Western blot to measure the expression of Runx2, Dlx-5,total Smad1/5/8,and phosphorylated Smad1/5/8.Finally,BMPR-Smad luciferase reporter assay was applied to measure the activation of BMPs/Smads signaling.Results BMP9 could induce the expression of COX-2 in C3H10T1/2 cells.Either inhibiting enzy-matic activity of COX-2 or knockdown of the expression of COX-2 reduced the BMP9 induced ALP activities in C3H10T1/2 cells,and COX-2 knockdown also inhibited the ectopic bone formation induced by BMP9 in C3H10T1/2 cells.Moreover,COX-2 knockdown inhibi-ted BMPR-Smad reporter activities and the phosphoryl-ation of Smad1/5/8,so did the expression of Smad6 and Smad7 .Conclusion COX-2 may play an impor-tant role in BMP9 induced osteogenic differentiation in MSCs by regulating the BMPs/Smads signaling trans-duction.

Objective To investigate the dosimetric performance of COMPASS system,a novel 3D quality assurance system for the verification of nasopharyngeal carcinoma volumetric modulated therapy (VMAT) treatment plan.Methods Eight VMAT treatment plans of nasopharyngeal carcinoma patients were performed with MasterPlan,a treatment planning system (TPS),and then these treatment plans were sent to the COMPASS and MOSAIQ system,a coherent control system,respectively.Comparison of the COMPASS reconstructed dose versus TPS dose was conducted by using the dose volume-based indices:dose received by 95％ volume of target ( D95％ ),mean dose ( Dmean ) and γ pass rate,dose to the 1％ of the spinal cord and brain stem volume ( D1％ ),mean dose of leaf and right parotid ( Dmean ),and the volume received 30 Gy for left and right parotid (V30).COMPASS can reconstruct dose with the real measured delivery fluence after detector commissioning.Results The average dose difference for the target volumes was within 1％,the difference for D95 was within 3％ for most treatment plans,and the γ pass rate was higher than 95％ for all target volumes.The average differences for the D1％ values of spinal cord and brain stem were ( 4.3 ± 3.0) ％ and ( 5.9± 2.9 ) ％ respectively,and the average differences for the Dmean values of spinal cord and brain stem were ( 5.3 ± 3.0 ) ％ and ( 8.0 ± 3.5 ) ％ respectively.In general the COMPASS measured doses were all smaller than the TPS calculated doses for these two organs.The average differences of the Dmean values of the left and right parotids were( 6.1± 3.1 ) ％ and ( 4.7 ± 4.4 ) ％ respectively,and the average differences of the V30 values of the left and right parotids were (9.4 ± 7.5 ) ％ and (9.4 ± 9.9)％ respectively.Conclusions An ideal tool for the VMAT verification,the patient anatomy based COMPASS 3D dose verification system can check the dose difference between the real delivery and TPS calculation directly for each individual organ,either target volumes or critical organs.

In this experimental study, the RGD-containing peptide was used to modify the surface of biomimetic PLGA-(ASP-PEG) matrix, and bone marrow stromal cells (BMSCs) were seeded onto these modified surfaces for three weeks. The effects of modified surfaces of matrix on the adhesion, proliferation and differentiation of BMSCs were explored. BMSCs were harvested from whole bone marrow of Sprague-Dawley (SD) rats in vitro, then were seeded onto peptide surface-modified matrix (Experiment group, EG) and matrices without modification (Control group, CG) respectively for three weeks. The number of adhesive cells was counted by using precipitation method after 4 h and 12 h incubation; the cells cytoskeletons were stained with FITC-conjugated phalloidin after 24h incubation; the cell density was investigated after 1 d, 2 d and 3 d of incubation; ALP activity of BMSCs was measured after 7 d, 14 d and 21 d of incubation with osteogenic medium. The cells from bone marrow were BMSCs and their purity was beyond 90% using flow cytometry (FCM) analysis. Sulphur binding energy in EG was shown by XPS to be 164 eV. BMSCs adhered on peptide surface-modified matrix were observed with SEM. Cell adhesion efficiency and quality in EG was better than that in CG, and cell cytoskeleton was more robust in EG. ALP activity was higher in EG than in CG. Peptide surface-modified PLGA-(ASP-PEG) was noted to have good compatibility with BMSCs and to promote cell adhesion and differentiation.
Animals ; Biocompatible Materials ; chemistry ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Adhesion ; drug effects ; Cell Differentiation ; drug effects ; Lactic Acid ; chemistry ; Oligopeptides ; chemistry ; Osteoblasts ; cytology ; Peptides ; chemistry ; Polyglycolic Acid ; chemistry ; Rats ; Rats, Sprague-Dawley ; Stromal Cells ; cytology ; drug effects ; metabolism ; Surface Properties ; Tissue Engineering ; Tissue Scaffolds

Neural stem cells (NSCs) cultured in vitro were implanted within three-dimensional (3D) and self-assembly hydrogel from IKVAV-containing peptide. The cytocompatibility of hydrogel with NSCs was explored. Neural cells harvested from the cerebral cortex of neonatal mice were dissociated mechanically and cultivated in serum-free media, and were immunohistochemically examined for Nestin, NSE (neuron specific enolase) and GFAP (Glial fibrillary acidic protein); the self-assembly hydrogel from 1 wt% amphiphilic peptide was formed with the addition of DMEM/F12 and observed under transmission electron microscope (TEM); 1 x 10(5) /ml NSCs implanted within hydrogel (3D culture system) and seeded onto the surface of coverslips covered with polylysine (two-dimensional culture system, 2D culture system) respectively were incubated without serum at 37 degrees C in 5% CO2 and 95% air. Then the cells in 3D culture system and 2D one were double-labeled with DAPI/Brdu and observed under fluorescence microscope. The Nestin-positive cells were found, and they were able to differentiate into NSE-positive neuron- like cells and GFAP-positive glial-like cells, TEM showed that the hydrogel derived from 1 wt% peptide solution was composed of nanofiber network with the fiber diameters ranging from 3 to 6 nm and the fiber length warying from 100 nm to 1.5 microm, NSCs labeled with Brdu in 2D system were thinly scattered, and those in 3D system formed many neurospheres; the positive rate in 3D was much higher than that in 2D system (P < 0.001). The self-assembly hydrogel from the amphiphilic peptide containing IKVAV sequence had good cyto-compatibility and promoted the proliferation of NSCs.
Animals ; Animals, Newborn ; Cells, Cultured ; Cerebral Cortex ; cytology ; Hydrogels ; pharmacology ; Materials Testing ; Neural Stem Cells ; cytology ; Peptides ; chemistry ; Rats ; Rats, Sprague-Dawley ; Surface-Active Agents ; chemistry

Aim To study the role of calcineurin signal transduction pathway in the anti-hypertrophic effect of L-arginine in vivo and in vitro.Methods The hypertrophic effects was assayed by calculating the right ventricular hypertrophy index(RVHI=right ventricle weight/left ventricle and septum weight),and atrial natriuretic peptide(ANP)mRNA expression in rat right ventricle hypertrophy model induced by monocrotaline(MCT) or by measuring the cell diameter,protein content,and ANP mRNA expression in hypertrophic cardiomyocyte induced by prostaglandin F2?(PGF2?).For mechanism studies,the intracellular free calcium concentration([Ca2+]i) in cultured cardiomyocytes was measured by using Fura 2/AM as a fluorescent indicator.ANP and CaN mRNA expressions,and expressions of CaN and its downstream effectors,NFAT3 and GATA4 proteins were assayed by RT-PCR and Western blot,respectively,in vivo and in vitro.Results In MCT-hypertrophic model,prevention-and treatment-administration of L-arginine,a nitric oxide(NO) precursor,200 mg?kg-1?d-1,could obviously inhibit the elevated RVHI and ANP mRNA expression;similar to that found in vivo.Addition of L-arginine 1 mmol?L-1 could markedly inhibit the increased cell diameter,protein content and the expression of ANP mRNA in the hypertrophic cardiomyocyte induced by PGF2? 100 nmol?L-1,and it could also decrease the elevated [Ca2+]i in vitro;notably,the above dose or concentration of L-arginine could blunt the elevated expressions of calcineurin mRNA and the calcineurin-,NFAT3-,GATA4-proteins induced by MCT or by PGF2?.These effects of L-arginine were blocked by NG-nitro-L-arginine-methyl ester,a NO synthase inhibitor,in vivo and in vitro.Conclusion These results suggest that calcineurin signal transduction pathway may play an important role in the NO-induced inhibition of cardiac hypertrophy.The anti-hypertrophic effects of L-arginine may involve the decrease of [Ca2+]i,and then inhibit the activated calcineurin pathway,through the release of NO.