1.In Vitro and in Vivo Evaluation of Scutellarin-phospholipid Complex Nanoemulsion and Analysis of Its Activity in Ameliorating LPS-induced Vascular Endothelial Injury
Tian LUO ; Zhiyong HE ; Xiangjun MAO ; Xue LIU ; Jinggang HE ; Yuan ZHI ; Xiangchun SHEN ; Qianli XU ; Ling TAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(10):159-168
ObjectiveTo evaluate some properties of scutellarin-phospholipid complex nanoemulsion(SCU-PC-NE), such as release, cell uptake and tissue distribution, and to investigate its effect on ameliorating lipopolysaccharide(LPS)-induced vascular endothelial injury. MethodSCU-PC-NE was prepared by weighting SCU-PC, ethyl oleate, Kolliphor HS15, 1,2-propylene glycol(50, 400, 514.3, 85.7 mg), respectively. And the appearance of SCU-PC-NE was observed by transmission electron microscope, the average paticle size and Zeta potential were measured by nanopotential particle size analyzer. The cumulative release of SCU-PC-NE in vitro was measured by dynamic dialysis, thiazolyl blue(MTT) colorimetric assay was used to investigate the effect of SCU-PC-NE on the viability of human umbilical vein endothelial cells(HUVECs), the inverted fluorescence microscope and flow cytometry were used to investigate cell uptake of HUVECs by SCU-PC-NE in vitro using coumarin 6 as a fluorescent probe, the tissue distribution of DiR/SCU-PC-NE labeled by near infrared fluorescent dyes was obeserved by small animal in vivo imaging system. The inflammation injury model was established by co-incubation with LPS(1 mg·L-1) and HUVECs, the effect of SCU-PC-NE on the levels of interleukin(IL)-1β and IL-6 were determined by enzyme-linked immunosorbent assay(ELISA), 18 Kunming male mice were randomly divided into blank group, model group, blank preparation group(equivalent to high dose group), SCU group and SCU-PC-NE low and high dose groups(5, 10 mg·kg-1), 3 mice in each group, and the drug administration groups were administered once in the tail vein at the corresponding dose every 48 h, equal volume of normal saline was given to the blank group and the model group, and the drug was administered for 4 consecutive times. Except for the blank group, the endothelial inflammatory injury was induced by intraperitoneal injection of LPS(10 mg·kg-1) at 12 h before the last administration in each group. Hematoxylin-eosin(HE) staining was used to investigate the effect of SCU-PC-NE on the histopathological changes in the thoracic aorta of mice. ResultThe appearance of SCU-PC-NE displayed pale yellow milky light, mostly spherical with rounded appearance and relatively uniform particle size distribution, with the average particle size of 35.31 nm, Zeta potential of 7.23 mV, and the encapsulation efficiency of 75.24%. The cumulative release in vitro showed that SCU-PC-NE exhibited sustained release properties compared with SCU. The cell viability of SCU-PC-NE was >90% at a concentration range of 1.05-8.4 mg·L-1. The results of cellular uptake experiments showed that the cellular uptake ability of SCU-PC-NE was significantly enhanced when compared with the SCU group(P<0.01). Compared with normal mice, the results of tissue distribution showed that the fluorescence intensity of DiR/SCU-PC-NE was significantly enhanced in the spleen, kidney, brain and thoracic aorta of mice at different time points after intraperitoneal injection of LPS(P<0.05, P<0.01), especially in thoracic aorta. ELISA results showed that the levels of IL-1β and IL-6 in the model group were significantly increased when compared with the blank group(P<0.05, P<0.01), and compare with the model group, all administration groups significantly down-regulated IL-1β level, with the strongest effect in the SCU-PC-NE high-dose group(P<0.01), and all administration groups significantly down-regulated IL-6 level, with the strongest effect in the SCU-PC-NE low-dose group(P<0.05). Compare with the blank group, the results of HE staining showed that the endothelial cells were damaged, the elastic fibers were broken and arranged loosely in the model group, although similar vascular injury could be observed in the blank preparation group, SCU group and SCU-PC-NE low-dose group, the vascular endothelial damage was significantly reduced in the high-dose group of SCU-PC-NE, which had a better effect than that in the SCU group. ConclusionSCU-PC-NE can promote the uptake of drugs by endothelial cells and effectively enriched in the site of vascular endothelial injury caused by LPS, suggesting that it has a protective effect on vascular endothelial injury and is a good carrier of SCU.
2.Role of MAPK Signaling Pathway in Prevention and Treatment of Rheumatoid Arthritis and Chinese Medicine Intervention: A Review
Huiming LI ; Shuangjuan BO ; Tao XING ; Guojun WEI ; Zhiyong WANG ; Xuemei TIAN ; Shenghua LI ; Tao LIU ; Yuguang LI ; Xiaotao WEI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(14):288-298
Rheumatoid arthritis (RA) is an autoimmune disease involving symmetrical small joints, with clinical manifestations such as small joint swelling, morning stiffness, progressive pain, and even joint deformity and loss of function. Due to the complex immune mechanism, the pathogenesis of RA remains unclear. However, studies have shown that the pathogenesis of RA is related to abnormal immune mechanism, increased synovial inflammatory response, abnormal biological behavior of RA fibroblast-like synoviocytes (RA-FLSs), and abnormal degradation of extracellular matrix. Mitogen-activated protein kinase (MAPK) plays a key role in the regulation of cell growth, proliferation, differentiation, and apoptosis. It is involved in the abnormal release and activation of inflammatory mediators in RA, the abnormal proliferation, migration, and invasion of RA-FLSs, synovial angiogenesis, bone erosion, and cartilage destruction. The thousands of years of practical experience show that Chinese medicine can effectively mitigate the clinical symptoms such as joint swelling, morning stiffness, and pain and delay the occurrence of joint deformity in RA patients. Moreover, the Chinese medicine treatment has the advantages of overall regulation, personalized treatment, multiple pathways and targets, high safety, few adverse reactions, and stable quality. Modern studies have confirmed that Chinese medicine can play a role in the prevention and treatment of RA by interfering in the MAPK signaling pathway, reducing pro-inflammatory cytokines, inhibiting the abnormal proliferation, migration, and invasion of RA-FLSs, regulating the apoptosis of RA-FLSs, and protecting extracellular matrix. This article elaborates on the key role of MAPK signaling pathway in the development of RA and reviews the latest research results of Chinese medicine intervention in MAPK signaling pathway for the prevention and treatment RA, aiming to provide a basis for the development of new drugs and the clinical application of Chinese medicine in the prevention and treatment of RA.
3.Diagnosis and treatment of chronic exertional compartment syndrome of the lower extremities: a review
Haofei WANG ; Xiaojun CHEN ; Tao WANG ; Junfei GUO ; Zhiyong HOU
Chinese Journal of Trauma 2024;40(3):266-274
Chronic exertional compartment syndrome (CECS) of the lower extremities is a common clinical condition characterized by exercise-induced pain in the extremities, which is predominantly observed in people who take an active part in sports, such as athletes. It is mainly presented as post-exercise pain in the lower extremities, probably accompanied by numbness and limb weakness, etc., affecting the patients′ life and work. The symptoms of CECS in the lower limbs are usually present after physical activities of a certain intensity, making them difficult to be identified through routine outpatient physical examination, and likely to be misdiagnosed and underdiagnosed. Furthermore, the absence of universally accepted and unified treatment standards for CECS of the lower extremities complicates the decision-making process regarding the necessity of surgical intervention and choice of surgical approach in the clinical practice. For this purpose, recent developments in the diagnosis and treatment of CECS of the lower extremities were reviewed to provide reference for its standardized diagnosis and treatment.
4.HIC Value of Mild Traumatic Rats under Anterior-Posterior and Lateral-Medial Craniocerebral Impact:An Equivalent Study
Guoxiang WANG ; Linna ZHU ; Xun WANG ; Qiuju CHEN ; Tao XIONG ; Qinghang LUO ; Jia YU ; Jingyu XU ; Zhiyong YIN ; Shengxiong LIU
Journal of Medical Biomechanics 2024;39(4):730-735
Objective To investigate the equivalent conversion of head injury criterion(HIC)under anterior-posterior(AP)and lateral-medial(LM)craniocerebral impact for mild craniocerebral injury in rats using motor evoked potential(MEP)and β-amyloid precursor protein(β-APP)immunohistochemistry(IHC).Methods Sixty healthy adult male SD rats were randomly divided into 0 m control group,0.5 m-AP and 0.5 m-LM injury groups,and 1 m-AP and 1 m-LM injury groups(12 rats in each group).The control group did not undergo any impact injury experiment.After the impact injury experiment,the injury and control groups were subjected to excessive anesthesia to produce β-APP immunohistochemical stained slices,and the percentage of positive area and integral optical density(IOD)in the brainstem pyramidal tract area of the slices were determined.The MEP groups were divided in the same manner as the IHC groups and the MEP amplitudes of the MEP and control groups were measured after the impact injury experiment.Results With an increase in the degree of injury,the decrease in MEP amplitude,percentage of positive areas,and IOD in the injury groups significantly increased.When the degree of injury was low,the sensitivity of IHC was higher than that of MEP.When the degree of injury was the same,the HIC in the LM direction was lower than that in the AP direction.When the HIC was the same,the degree of injury in the LM direction was greater than that in the AP direction.Conclusions The joint evaluation of MEP and β-APP can provide experimental references for the study of HIC equivalent conversion in AP-LM craniocerebral impact injury.
5.Research on localization practice of the clinical application integration course in sino-foreign cooperative clinical medicine program
Diansa GAO ; Tao LUO ; Zhiyong ZHANG ; Xuan ZHANG ; Que ZHU ; Guobin YIN ; Zhen QUAN
Chinese Journal of Medical Education Research 2024;23(10):1363-1368
Among the current curricula of medical teaching in China, most courses focus on the integrated teaching of a single organ-system combination, while there are relatively a few integrated courses that focus on the multiple dimensions between different organs and systems and between medical sciences and social sciences. In 2016, Chongqing Medical University started to cooperate with University of Leicester to establish the clinical medicine major and introduced the course of Integration for Clinical Application (ICA) that had been run well in University of Leicester for years. With reference to the education goal of our university, the curriculum group adopted a series of actions for the localization of this course from the aspects of teaching objectives, contents, teaching model, education resources, and quality of faculty. After the completion of the first round of this course, the passing rate reached 86.96%(100/115) in the quantified evaluation of accomplishment, which was higher than the passing rate of other courses introduced from University of Leicester. The quantitative expert assessment of this course also ranked among the top courses in our university, and student assessment showed that the ability indicators were improved by 25.00%- 38.00%. The above data show that good results have been achieved for the curriculum localization of ICA.
6.Exploration of the Application of Blockchain Technology in the Future Medical Service System
Huan WANG ; Tao LIU ; Jing WANG ; Yanan GAO ; Xin LI ; Liyuan HU ; Zihang FENG ; Zhiyong ZHAO ; Qingyang HUANG ; Quanyu MENG ; Xiaojuan ZHANG ; Hexuan CUI ; Tianming CHEN
Journal of Medical Informatics 2024;45(11):91-94,103
Purpose/Significance To explore the feasibility of applying blockchain technology to the current healthcare system of hos-pitals,and to achieve the purpose of protecting patients'privacy to the greatest extent possible at a lower cost.Method/Process 505 questionnaires are randomly distributed and collected from people of different age groups in Beijing,Tianjin,Shanghai and Shenzhen who have a certain degree of understanding of blockchain technology,and the results are analyzed.Result/Conclusion Different age groups are highly concerned about personal privacy and privacy protection,and are willing to accept blockchain as an emerging technology.There is a greater demand and acceptance for the application of blockchain technology in the primary health care systems.
7.Research progress on alterations in host signaling pathways triggered by porcine parvovirus infection
Min ZHOU ; Deyuan TANG ; Zhiyong ZENG ; Bin WANG ; Tao HUANG ; Wenwen HU ; Yinming MAO ; Piao ZHOU ; Song HE
Chinese Journal of Veterinary Science 2024;44(10):2294-2300
Porcine parvovirus(PPV)is a causative agent of porcine parvovirus infection(PPI),which significantly impacts the pig industry due to its association with reproductive dysfunction.The condition is characterized by stillbirth,mummified fetuses,fetal weakness,and abortion in pregnant sows.The pathogenic mechanism remains incompletely understood,with no effective therapeutic drugs available currently.Despite extensive research on the host's response to PPV in-fection and identification of various signaling pathway transduction mechanisms,a comprehensive understanding is still lacking.This review aims to summarize the current knowledge regarding al-terations in related signaling pathways following PPV infection,provide novel insights into the pathogenesis of PPV and facilitate the drug development.
8.Research progress of immune response mechanisms and prevention and control of porcine circovirus type 2
Yinming MAO ; Deyuan TANG ; Zhiyong ZENG ; Bin WANG ; Tao HUANG ; Song HE ; Piao ZHOU ; Zhengbo LIAO ; Shenglin YUAN ; Xu CHEN
Chinese Journal of Veterinary Science 2024;44(11):2483-2489
Porcine circovirus type 2(PCV2)is the main pathogen causing porcine circovirus related diseases.PCV2 infection in pigs may lead to porcine dermatitis and nephrotic syndrome(PDNS)and weaned piglets multiple system failure syndrome(PMWS),etc.At present,the pathogenic mechanism is not fully understood.PCV2 is a single strand of negative link DNA,which can cause immune suppression in the body and lead to increased secondary susceptibility,which has a syner-gistic effect with various pig diseases and brings major economic losses to the pig industry.Al-though there are commercial vaccines,the prevention of vaccines has certain limitations and there is no effective drug treatment so far,an outbreak will threaten people's life and health and public safety,resulting in significant economic losses.In order to understand the latest progress of PCV2 escape mechanism and prevention and control,this paper summarizes the inhibition of interferon production,regulation of apoptosis,regulation of autophagy,regulation of pyroptosis and inflam-matory response,evasion of adaptive immune response,and prevention and control of PCV2,in or-der to provide new theoretical ideas for the research and prevention and control of PCV2.
9.Effective dose and safety of ciprofol in combination with butorphanol for painless gastroscopy in elderly patients
Zhiyong FANG ; Xiaojuan QIU ; Tao FENG ; Qingming BIAN
Journal of Clinical Medicine in Practice 2024;28(12):82-85
Objective To investigate the median effective dose (ED50) and 95% effective dose (ED95) of ciprofol in combination with butorphanol for painless gastroscopy in elderly patients. Methods A total of 27 elderly patients with selective gastroscopy were conducted with intravenous induction of 5 μg/kg butorphanol for at least 30 seconds, and 3 minutes later, they were administered intravenously with ciprofol for at least 30 seconds; gastroscopy was performed when the eyelash reflex disappeared or the Modified Observer's Assessment of Alertness/Sedation (MOAA/S) score was less than one point. The initial dose of ciprofol was set as 0.2 mg/kg, and the modified sequential allocation method was used; if there was a positive response during the insertion of the gastroscope, the dose for the next patient would increase by one level, and vice versa, the dose would decrease by one level, with an adjacent dose concentration gradient of 0.05 mg/kg. A positive response to gastroscope insertion was defined as coughing, swallowing, or limb movements when the gastroscope entered the throat. The study was terminated after 7 reversals in ciprofol dose adjustments. ED50, ED95, and corresponding 95% confidence intervals (95%CI) of ciprofol combined with butorphanol for painless gastroscopy were calculated. Results When combined with butorphanol, the ED50 of ciprofol for painless gastroscopy in elderly patients was 0.253 mg/kg (95%CI, 0.219 to 0.290), and the ED95 was 0.328 mg/kg (95%CI, 0.290 to 0.521). Conclusion When combined with butorphanol, the ED50 of ciprofol for painless gastroscopy in elderly patients is 0.253 mg/kg, and the ED95 is 0.328 mg/kg.
10.Study on the mechanism of VEGF inducing tolerogenic dendritic cells in oral squamous cell carcinoma
Tao XIAO ; Yijia HE ; Yaoping ZHU ; Fengyao HAO ; Yan CHEN ; Zhiyong WANG
STOMATOLOGY 2023;43(3):204-211
Objective:
To investigate the mechanism of vascular endothelial growth factor(VEGF) inducing tolerogenic dendritic cells(DCs) in oral squamous cell carcinoma (OSCC).
Methods:
The DCs were divided into four groups: Control group (DC), VEGF group (VEGF added into DC), Co-culture group (DC co-cultured with SCC7) and Anti-VEGF group (anti-VEGF antibody added into DC co-cultured with SCC7). Flow cytometry (FCM) was used to detect DC surface markers. To detect the effect of DC on proliferation activity of T lymphocyte, the experiment included five groups: Nc group (T lymphocyte), Control group (T lymphocyte added into DC), VEGF group (T lymphocyte + DC + VEGF), Co-culture group (T lymphocyte + DC + supernatant of SCC7) and Anti-VEGF group (T lymphocyte + DC + supernatant of SCC7 + anti-VEGF antibody). Subsequently, the mixed lymphocyte reaction(MLR) was conducted. The expression levels of indole-2, 3-doxygenase(IDO)and programmed cell death 1 ligand 1(PD-L1)in DC were detected by western blot, real time PCR and FCM respectively. For the cytotoxic lymphocyte (CTL) assay, SCC7 cells and CTLs were mixed and CTL-mediated SCC7 cells cytotoxicity was tested. The experiment included four groups: Control group (T lymphocyte + DC), IDO inhibition group (T lymphocyte + DC + IDO inhibitor), Anti-PD-L1 antibody group (T lymphocyte + DC + anti-PD-L1 antibody) and Combination group (T lymphocyte + DC + IDO inhibitor + anti-PD-L1 antibody). The SCC7 tumor-bearing mice treated with IDO inhibitor and the anti-PD-L1 antibody were sacrificed and the tumor inhibition rate and the spleen index were determined.
Results:
Compared with Control group, exogenous VEGF or SCC7 co-culture inhibited the relative number of DC expressing CD11C, CD80, CD86, CD40 and MHC Ⅱ. The positive DCs were increased in the Anti-VEGF group compared with VEGF or Co-culture group. In VEGF or Co-culture group, the number of T cells stimulated by SCC7-pulsed DCs was decreased compared with Control group. However, the ability of Anti-VEGF group to induce T cell proliferation was significantly increased compared with VEGF or Co-culture group. Significantly increased expression of IDO and PD-L1 were observed in VEGF and Co-culture group. However, this was partially reversed by addition of anti-VEGF antibody into the co-culture system. Compared with Control group, the expressions of CD11C and CD86 in DC in both the IDO inhibition group and Anti-PD-L1 antibody group were increased, and were significantly higher in the Combination group compared with the single drug groups. The similar results were exhibited in MLR and CTL assay. In vivo, the results revealed that the tumors obtained from the mice in three experimental groups were smaller than those in the control group. Furthermore, the tumor volume of the Combination group was the smallest. The spleen index of each group was calculated and the results showed the spleen index of the three experimental groups was significantly higher than that of Control group.
Conclusion
VEGF in OSCC micro-environment inhibits the maturation and function of DC that are transformed into tolerogenic DC by high expression of IDO and PD-L1.


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