1.Exploration of the Pharmacological Substance Basis and Action Characteristics of Wenxing Jingjintong Gel Patch in the Treatment of Rheumatoid Arthritis Based on System Pharmacology and LC-MS/MS Technology
Anyu SU ; Weibin HUANG ; Jingyuan MO ; Zixuan ZHANG ; Xun XIE ; Xiaobing HUANG ; Bilian ZHONG ; Yongping ZHANG ; Lisheng WANG ; Chunyi WANG
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(5):706-718
Objective To explore the key active ingredients and action characteristics of Wenxing Jingjintong Gel Patch in the treatment of rheumatoid arthritis(RA)based on the systematic pharmacology and LC-MS/MS technology.Methods The information of active ingredient from Wenxing Jingjintong Gel Patch was established through LC-MS/MS analysis and literature retrieval.The targets of the active ingredients were predicted using Swiss Target Prediction platform and then mapped with the RA-related targets obtained from GeneCards,DrugBank,and OMIM databases to identify the intersecting targets.The"active ingredients-effective targets"network was constructed through the Cytoscape software.The shared targets were imported into STRING database to construct a protein-protein interaction network.GO function and KEGG pathway enrichment analysis were performed using the Metascape database.Molecular docking studies were conducted using AutoDock software to investigate the interactions between key ingredients and target proteins.Results A total of 142 active ingredients were identified in Wenxing Jingjintong Gel Patch by wsing LC-MS/MS,which were further supplemented to 174 through literature retrieval.There were 175 shared targets between the active ingredients and RA.It was anticipated that Wenxing Jingjintong Gel Patch exerted immune regulation and anti-inflammatory and analgesic effects through the interaction between key active ingredients such as berberine,neobavaisoflavone,and palmatine chloride with key targets,including TNF,IL6,and AKT1 to regulate PI3K/Akt1,JAK/STAT,and MAPK signaling pathways.In 1 152 molecular docking validation,94%of them had binding energies less than-5.0 kcal·mol-1,while 51%of them had binding energies less than-7.0 kcal·mol-1.It was indicated that there was a good binding affinity between the potential active ingredients and core targets.Conclusion This study predicted the active ingredients and action characteristics of Wenxing Jingjintong Gel Patch in the treatment of RA,which provided a theoretical basis for further clinical application and quality control.
2.Preparation,Characterization,and In Vitro Evaluation of Dissolvable Microneedles Loaded with Hyaluronic Acid-modified Sinomenine Hydrochloride Liposome
Xun XIE ; Nijun ZHONG ; Jingyuan MO ; Xiaoling HUANG ; Jiaxiang ZOU ; Yongping ZHANG ; Lisheng WANG
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(9):1428-1436
Objective To prepare a dissolvable microneedle(DMN)with a tip-layer loaded with hyaluronic acid(HA)modified sinomenine hydrochloride liposomes(HA-SMH-Lip),as well as characterize,evaluate its in vitro transdermal permeability,cellular uptake ability,and anti-inflammatory ability.Methods HA-SMH-Lip-DMNs were prepared by a two-step casting method,and the drug loading capacity was determined using HPLC.The morphology,skin permeation properties and in vitro transdermal ability were investigated by scanning electron microscopy,puncture assay and Franz diffusion cell method.Fluorescent microneedles were prepared by replacing HA-SMH-Lip with fluorescein isothiocyanate liposomes(HA-FITC-Lip/FITC-Lip).The uptake behavior of inflammation cells on HA-FITC-Lip-DMNs/FITC-Lip-DMNs was investigated using a flow cytometer and a fluorescence microscope.To evaluate the anti-inflammatory activity of HA-SMH-Lip-DMNs,the levels of inflammatory factors including nitric oxide(NO),tumor necrosis factor α(TNF-α),interleukin 1β(IL-1β),and IL-10 in cell supernatants were measured using an ELISA kit.Results The prepared HA-SMH-Lip-DMNs have uniform shape and size,integral and visually pleasing array,and an average drug loading of(114.01±1.04)μg.Additionally,they have good puncture ability.The results of in vitro transdermal experiments showed that the accumulated amounts of HA-SMH-Lip-DMNs were(101.47±2.91)μg·cm-2 at 36 hours.Its transdermal ability was better than that of the SMH solution group and SMH liposome group.In vitro cellular uptake results indicated that HA-FITC-Lip-DMNs were more effectively taken up by RAW 264.7 cells(P<0.01).Compared to the model group,HA-SMH-Lip-DMNs group significantly reduced TNF-α,IL-1β,and NO levels while increase IL-10 levels(P<0.01).Conclusion The prepared HA-SMH-Lip-DMNs have a complete and beautiful morphology with excellent cellular uptake capability,remarkable in vitro transdermal performance,and potent anti-inflammatory properties.HA-SMH-Lip-DMNs are expected to become a new type of transdermal drug delivery system.
3.Value of two-dimensional ultrasound in evaluating the medial head of gastrocnemius in patients with decreased muscle mass
Zecheng WANG ; Guorong LYU ; Huohu ZHONG ; Zhenhong XU ; Lisheng YAN
Chinese Journal of Ultrasonography 2022;31(11):978-983
Objective:To analyze the correlation between the muscle thickness (MT) and echo intensity (EI) of the medial head of gastrocnemius and muscle mass, strength and functional parameters, and to evaluate the diagnostic efficacy of MT and EI for decreased muscle mass.Methods:Fifty-two subjects who visited the Department of Ultrasound Medicine of the Second Affiliated Hospital of Fujian Medical University from March to December 2021 were collected.MT of the medial head of gastrocnemius was measured by two-dimensional ultrasound, and EI values in the region of interest were analyzed using Image J software. Twenty of the subjects were selected to assess the intra-observer and inter-observer agreements in measuring MT and EI values using the intraclass correlation coefficient (ICC) and Altman-Bland analysis was performed. The subjects were divided into healthy control group (30 cases) and decreased muscle mass group (22 cases) according to 2019 Asian Sarcopenia Working Group (AWGS) diagnostic criteria, the differences in MT and EI values between the two groups were compared, and the correlation between MT, EI values and muscle mass, strength, function were analyzed using Pearson correlation analysis. The diagnostic cutoff values of MT and EI values for decreased muscle mass were obtained using ROC curves, and the diagnostic efficacy was compared.Results:①The ICC of MT and EI in medial head of gastrocnemius detected by the same physician and different physicians were greater than 0.97, and the retest reliability was good. ②Compared with the healthy control group, the MT value in medial head of gastrocnemius decreased and the EI value increased in the decreased muscle mass group ( t=4.759, -3.789; all P<0.001). ③The MT value in medial head of gastrocnemius was positively correlated with appendicular skeletal muscle mass (ASM), appendicular skeletal muscle mass index (ASMI), lower limb skeletal muscle mass (LLSM), grip strength, and gait speed ( r=0.579, 0.568, 0.585, 0.495, 0.415; all P<0.001), of which, the correlation with LLSM was the best; ④EI in medial head of gastrocnemius was negatively correlated with ASM and ASMI, LLSM, grip strength, and gait speed, ( r=-0.583, -0.591, -0.590, -0.537, -0.455; all P<0.001), and among which the correlation with ASMI was the best. ⑤The medial head of gastrocnemius MT and EI were 14.33 mm and 58.85 for predicting decreased muscle mass, respectively, and there was no significant difference in the diagnostic performance of the two for decreased muscle mass ( Z=0.734 3, P>0.05). Conclusions:The MT and EI values of the medial head of gastrocnemius can accurately assess muscle mass and can be used in the evaluation of patients with decreased muscle mass.
4.Identification of Medium-Length Antineurofilament Autoantibodies in Patients with Anti-N-Methyl-D-Aspartate Receptor Encephalitis
Shisi WANG ; Cancan XU ; Xiaobo SUN ; Yifan ZHOU ; Yaqing SHU ; Shangzhou XIA ; Zhengqi LU ; Wei QIU ; Xiaofen ZHONG ; Lisheng PENG
Journal of Clinical Neurology 2020;16(3):470-479
Background:
and Purpose: Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is a severe central nervous system disorder mediated by NMDAR antibodies that damages neurons. We investigated the correlation between cytoskeletal autoantibodies and the clinical severity in patients with anti-NMDAR encephalitis.
Methods:
Non-NMDAR autoantibodies were identified by screening matched cerebrospinal fluid (CSF) and the serum samples of 45 consecutive patients with anti-NMDAR encephalitis and 60 healthy individuals against N-methyl-D-aspartate receptor 1-transfected and nontransfected human embryonic kidney 293T cells. Immunocytochemistry was performed to assess antibody binding in rat brain sections and primary cortical neurons. Cell-based assays and Western blotting were applied to identify autoantibodies targeting medium neurofilaments (NFMs). We compared clinical characteristics between patients with NMDAR encephalitis who were positive and negative for anti-NFM-autoantibodies.
Results:
Anti-NFM autoantibodies were detected in both the serum and CSF in one patient (2%) and in the serum only in six patients (13%). No antibodies were detected in the serum of healthy controls (7/45 vs. 0/60, p=0.0016). Four of the seven patients with anti-NFM autoantibodies in serum were children (57%), and three (43%) had abnormalities in brain magnetic resonance imaging. These patients responded well to immunotherapy, and either no significant or only mild disability was observed at the last follow-up. Anti-NMDAR encephalitis did not differ with the presence of anti-NFM autoantibodies.
Conclusions
Anti-NFM autoantibodies may be present in patients with anti-NMDAR encephalitis, indicating underlying neuronal damage. A large cohort study is warranted to investigate the clinical differences between patients with NMDAR encephalitis according to their antiNFM antibody status.
5. Analysis on key points for construction of trauma emergency center of Jiangxi Province
Yuanlin ZENG ; Haiming CHEN ; Lisheng LUO ; Xianlai XU ; Haigang XU ; Zhili LIU ; Sheng LIU ; Bin FU ; Xuefeng HUANG ; Zhongping YUAN ; Lidong WU ; Yuhua WAN ; Youjia TANG ; Chunming HUANG ; Peng RAO ; Hongfa ZHONG ; Bohe LI ; Yongan ZHANG ; Jiahua TANG ; Bo YOU
Chinese Journal of Trauma 2019;35(12):1126-1129
Trauma is the leading cause of death for people under 40 years old in the world. At present, the rescue and treatment system of trauma patients in China is not yet well established, and the mortality of trauma patients is higher than those in the developed countries. Improving the treatment system is the key to reducing the trauma mortality. In order to innovate the service mode of trauma first aid, further promote the establishment of regional trauma first aid system, improve the ability of trauma treatment, reduce the mortality and disability rate of trauma patients in Jiangxi Province, recently Health Commission of Jiangxi Province and the First Affiliated Hospital of Nanchang University have reached a consensus on the establishment of Jiangxi trauma first aid center. In order to provide reference for the construction of trauma treatment system, the author analyzes the following aspects including functional positioning, basic requirements, organization management, and evaluation of core indicators.
6.Phenotypic characteristics of LAP+CD4+T lymphocytes in colorectal cancer tissues
Wu ZHONG ; Zhiyuan JIANG ; Shibiao ZHONG ; Leichang ZHANG ; Jiahao HUANG ; Sen ZHANG ; Lisheng CHEN ; Yunfei CAO
Chinese Journal of Oncology 2016;38(8):596-601
Objective To analyze the phenotypic characteristics of LAP+CD4+ T lymphocytes and investigate their molecular mechanisms in colorectal cancer ( CRC ) microenvironment. Methods Fifty colorectal cancer patients treated in our two hospitals from January 2014 to May 2014 were included in this study. Their tumor tissues and adjacent normal tissues, peripheral blood samples, and peripheral blood samples of 25 healthy donors ( HD) were collected to isolate the lymphocytes. The different expressions of CCR7, CD45RA, Foxp3, CTLA?4, CCR4 and CCR5 in LAP+CD4+ T and LAP-CD4+ T lymphocytes were analyzed by flow cytometry ( FCM) . Results The FCM assay detected that the percentage of LAP+CD4+ T cells in peripheral blood of the CRC patients were significantly higher than that of HD [(9.44±3.18)%versus (1.49±1.00)%, P<0.001]. In addition, significantly more LAP+CD4+ T cells were also recruited into tumor tissue than those in the tumor?adjacent normal tissue [(11.76±3.74)% versus (3.87±1.64)%, P<0.001]. LAP+CD4+ T cells in the tumor?adjacent normal tissue and peripheral blood of both HDs and CRC patients mainly displayed a central memory phenotype. However, effector memory lymphocytes were predominant in the tumor tissue.In the tumor tissue, the expression of Foxp3 in the LAP+CD4+ T cells was (3.87±1.12)%, significantly lower than that in the LAP-CD4+ T cells (16.70±2.61)%, (P<0.001); the expression of CTLA?4 in the LAP+CD4+ T cells was (36.36±19.14)%, significantly higher than the (19.60± 8.91)% in the LAP-CD4+ T cells ( P<0. 001); the expression of CCR4 in the LAP+CD4+ T cells was (37.72±11.14)%, significantly higher than the (30.06±9.14)% in the LAP-CD4+ T cells (P<0.001);and the expression of CCR5 in the LAP+CD4+ T cells was (18.86±7.10)%, significantly higher than the (13.92±3.31)% in the LAP-CD4+ T cells (P<0.001). Conclusions LAP+CD4+ T cells with low expression of Foxp3 and high expressions of CTLA?4, CCR4 and CCR5 are tend to be enriched and accumulated in the tumor tissue. The unique phenotypic characteristics make these cells a distinct subset of lymphocytes, apparently different from the traditional CD4+CD25+ Treg cells.
7.Phenotypic characteristics of LAP+CD4+T lymphocytes in colorectal cancer tissues
Wu ZHONG ; Zhiyuan JIANG ; Shibiao ZHONG ; Leichang ZHANG ; Jiahao HUANG ; Sen ZHANG ; Lisheng CHEN ; Yunfei CAO
Chinese Journal of Oncology 2016;38(8):596-601
Objective To analyze the phenotypic characteristics of LAP+CD4+ T lymphocytes and investigate their molecular mechanisms in colorectal cancer ( CRC ) microenvironment. Methods Fifty colorectal cancer patients treated in our two hospitals from January 2014 to May 2014 were included in this study. Their tumor tissues and adjacent normal tissues, peripheral blood samples, and peripheral blood samples of 25 healthy donors ( HD) were collected to isolate the lymphocytes. The different expressions of CCR7, CD45RA, Foxp3, CTLA?4, CCR4 and CCR5 in LAP+CD4+ T and LAP-CD4+ T lymphocytes were analyzed by flow cytometry ( FCM) . Results The FCM assay detected that the percentage of LAP+CD4+ T cells in peripheral blood of the CRC patients were significantly higher than that of HD [(9.44±3.18)%versus (1.49±1.00)%, P<0.001]. In addition, significantly more LAP+CD4+ T cells were also recruited into tumor tissue than those in the tumor?adjacent normal tissue [(11.76±3.74)% versus (3.87±1.64)%, P<0.001]. LAP+CD4+ T cells in the tumor?adjacent normal tissue and peripheral blood of both HDs and CRC patients mainly displayed a central memory phenotype. However, effector memory lymphocytes were predominant in the tumor tissue.In the tumor tissue, the expression of Foxp3 in the LAP+CD4+ T cells was (3.87±1.12)%, significantly lower than that in the LAP-CD4+ T cells (16.70±2.61)%, (P<0.001); the expression of CTLA?4 in the LAP+CD4+ T cells was (36.36±19.14)%, significantly higher than the (19.60± 8.91)% in the LAP-CD4+ T cells ( P<0. 001); the expression of CCR4 in the LAP+CD4+ T cells was (37.72±11.14)%, significantly higher than the (30.06±9.14)% in the LAP-CD4+ T cells (P<0.001);and the expression of CCR5 in the LAP+CD4+ T cells was (18.86±7.10)%, significantly higher than the (13.92±3.31)% in the LAP-CD4+ T cells (P<0.001). Conclusions LAP+CD4+ T cells with low expression of Foxp3 and high expressions of CTLA?4, CCR4 and CCR5 are tend to be enriched and accumulated in the tumor tissue. The unique phenotypic characteristics make these cells a distinct subset of lymphocytes, apparently different from the traditional CD4+CD25+ Treg cells.
8.Diagnosis and treatment of type Ⅰ congenital biliary dilatation
Shibiao ZHONG ; Wu ZHONG ; Leichang ZHANG ; Lisheng CHEN
Chinese Journal of Hepatobiliary Surgery 2015;21(3):213-216
Type Ⅰ congenital biliary dilatation is a congenital biliary duct defect disease.It is also called congenital choledochocele cyst (CCC).Endoscopic ultrasonography (EUS),magnetic resonance cholangiopacreatography (MRCP) and endoscopic retrograde cholangiopancreatography (ERCP) are very important for the diagnosis.Operation is generally regaded as the best choose.It is recommended that cholecystectomy + choledochal cyst excision + hepatic duct jejunum Roux-Y anastomosis is the chief therapeutic method.This article reviews the pathogenesis,classifications,diagnosis and treatment of the disease.
9.Survival analysis of 345 patients with colorectal cancer undergoing palliative resection
Shibiao ZHONG ; Wu ZHONG ; Leichang ZHANG ; Lisheng CHEN
Chinese Journal of Digestive Surgery 2015;14(6):470-474
Objective To investigate the related factors affecting prognosis of patients with colorectal cancer undergoing palliative resection.Methods The clinical data of 345 patients with colorectal cancer who underwent palliative resection at the First Affiliated Hospital of Guangxi Medical University between January 2007 and December 2011 were retrospectively analyzed.Patients selected the emergent operations or restrictive operation based on the conditions of patients,and then received chemotherapy,radiotherapy and biotherapy.Chemotherapy regimens included FOLFOX4 (5-FU/CF + oxaliplatin) regimen,XELOX (capecitabine + oxaliplatin) regimen and FOLFIRI (5-FU + CF + irinotecan) regimen.Biotherapy regimens included molecular targeted therapies using bevacizumab and cetuximab.The follow-up was applied to patients by outpatient examination,telephone interview and correspondence once every 3 months up to March 1,2014.The survival curve was drawn by the Kaplan-Meier method.The survival rate was analyzed using the Log rank test.The multivariate analysis was done using the COX regression model.Results Of the 1 930 patients,345 patients received palliative resection with a percentage rate of 17.876%.Among the 345 patients,104 patients received emergent operations and others received restrictive operations.After operation,178 patients received adjuvant treatment,FOLFOX4 regimens were done on 85 patients,XELOX regimens on 32 patients,FOLFIRI regimens on 20 patients,radiotherapy on 17 patients and molecular targeted therapies on 24 patients.Seven patients died in the perioperative period and other patients were followed up for 3.0-82.0 months with a median time of 14.0 months.The 1-,3-and 5-year survival rates and the mean survival time were 49.57%,11.88%,6.38% and 22.6 months,respectively.The results of univariate analysis showed that the CEA levels,bowel obstruction,primary tumor resection,peritoneal implantation,distal metastasis,lymph node metastasis,tumor differentiation,postoperative adjuvant therapy were related factors affecting the prognosis of patients undergoing palliative resection (x2=3.742,18.795,37.641,13.470,4.228,5.835,4.108,6.875,P <0.05).The results of multivariate analysis showed that the bowel obstruction,without primary tumor resection,peritoneal implantation,low-differentiation of tumor and without postoperative adjuvant therapy were the independent risk factors affecting the prognosis of patients undergoing palliative resection (RR =1.674,2.273,1.947,1.582,1.342,95% confidence interval:1.193-2.485,1.646-4.376,1.497-3.587,1.184-2.382,1.032-2.074,P < 0.05).Conclusion The low-differentiation of tumor,peritoneal implantation,bowel obstruction,without primary tumor resection and without postoperative adjuvant therapy are the independent risk factors affecting the prognosis of patients with colorectal cancer undergoing palliative resection.
10.Effects of Cdc20 mutation on growth of mouse embryonic fibroblast
Juling FENG ; Lei ZHAO ; Juan XIE ; Mingshu MO ; Qingjun GUI ; Yong YOU ; Hui ZHONG ; Lisheng WANG
Acta Anatomica Sinica 2014;(3):383-387
Objective Investigation of biological characteristics of Cdc 20AAA/+APCmin/+ mouse embryonic fibroblast(MEFs) indicate the effect of Cdc20AAA/+on growth of mouse embryonic fibroblast and the possible mechanism . Methods MEFs of Cdc20AAA/+APCmin/+, Cdc20AAA/+, APCmin/+ and WT genotype were harvested from embryos for analysis.The growth characteristics of Cdc20AAA/+APCmin/+, Cdc20AAA/+,APCmin/+and WT mouse embryonic fibroblast were analyzed through growth curve analysis and foci formation assay .Separation of sister chromatid and the presence of aneuploid were detected by karyotype analysis .Results Cell proliferation assays showed that Cdc 20AAA/+APCmin/+cells grew at an accelerated rate compared with APC min/+MEFs(P<0.01).Foci formation assay showed that the clone forming ability was significantly increased .Cdc20AAA/+APCmin/+MEFs showed a significant increase in the frequency of aneuploid compared with WT MEFs , which had a karyotype of 38 and contained prematurely separated sister chromatids .Conclusion Cdc20 carrying a null allele (Cdc20AAA/+) may accelerate the growth and proliferation of APC min/+MEFs and present the growth characteristics of the tumor cells .The possible mechanism may be associated with chromosome instability .

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