1.Construction of glioma microfluidic chip model and its application research on evaluation the medicinal efficacy of the Chinese medicine Scutellaria barbata
Piaoxue YOU ; Lan CHEN ; Yiwei SHI ; Hui WANG ; Liang CHAO ; Zhanying HONG
Journal of Pharmaceutical Practice and Service 2025;43(2):59-66
Objective To construct a glioma microfluidic chip model to simulate tumor microenvironment for evaluating the medicinal efficacy of anti-glioma traditional Chinese medicines. Methods Glioblastoma cells U251 were seeded into microfluidic chips with different culture modes, and the cell viability and tumour microenvironment within the constructed model were characterized. Fluorescence staining was used to evaluate the effects of the positive drugs temozolomide (TMZ) and docetaxel (DOC) on the cell activity and apoptosis within the model, which was applied to evaluate the medicinal efficacy of the extracts of the herb Scutellaria barbata on gliomas. Results The cells in the constructed U251 microfluidic chip model displayed high viability and were able to mimic the hypoxic microenvironment of tumor to a certain extent. The viability of the U251 cells in the microfluidic chips decreased with the increasing of the concentration of the positive drug, and the viability of the 3D cultured U251 cells was higher than that in the 2D condition (P<0.05). The intracellular mitochondrial membrane potential decreased with the increasing of the concentration of the positive drug. And the 2 mg/ml Scutellaria barbata extract killed U251 cells to a certain extent and reduced the mitochondrial membrane potential of the cells in the model. Conclusion This study successfully constructed a microfluidic chip model of glioma that could effectively simulate the tumor microenvironment and rapidly evaluate the anti-tumor medicinal efficacy, which provided a new strategy for the medicinal efficacy evaluation and active components screening of anti-glioma traditional Chinese medicines.
2.Effect of Yiqi Huoxue Tongluo Decoction on miR-126a-5p and VEGF signaling pathway in cervical spondylotic myelopathy model rats
Dan LIU ; Zhanying TANG ; Pan LI ; Weina YUAN ; Fangfang LI ; Qian CHEN ; Zhijun HU
Tianjin Medical Journal 2024;52(3):273-277
Objective To investigate the effect of Yiqi Huoxue Tongluo Decoction on microRNA-126a-5p(miR-126a-5p)and vascular endothelial growth factor(VEGF)signaling pathway in cervical spondylotic myelopathy model rats.Methods Thirty healthy male SD rats were divided into the sham operation group,the model group and the traditional Chinese medicine(TCM)group by random number table method.Cervical spondylotic myelopathy models were prepared in the model group and the TCM group.The TCM group was given intragastric administration of Yiqi Huoxue Tongluo Decoction,while the sham operation group and the model group were given intragastric administration of normal saline for 12 weeks.After intervention,the threshold of mechanical stimulation and retraction time of thermal stimulation in each group were measured by behavior tests.Rats were sacrificed to collect intervertebral disc tissue for hematoxylin-eosin(HE)staining and observe the number of vascular buds in intervertebral disc.Rat intervertebral disc annulus fibrosus cells were subjected to terminal dexynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)staining.The miR-126a-5p and VEGF mRNA of rat intervertebral disc tissue were detected by real-time fluorescence quantitative polymerase chain reaction(RT-PCR).The expression of VEGF protein of rat intervertebral disc tissue was detected by Western blot assay.Results Compared with the sham operation group,the number of vascular buds in intervertebral disc was decreased in the model group and the TCM group.The cell destruction of intervertebral disc annulus was obvious in rats,and apoptosis was high and cell density decreased.Mechanical stimulation threshold decreased,and mechanical stimulation threshold decreased.The level of miR-126a-5p was decreased,and the expression levels of VEGF mRNA and protein were increased.Compared with the model group,the number of vascular buds in intervertebral disc was increased in the TCM group.The destruction of intervertebral disc annulus cells was alleviated in rats.The apoptosis of annulus fibrosus cells in intervertebral disc decreased and cell density increased.The threshold of mechanical stimulation increased,and the retraction time of thermal stimulation was prolonged.The level of miR-126a-5p increased,and the expression levels of VEGF mRNA and protein decreased(P<0.05).Conclusion The mechanism of Yiqi Huoxue Tongluo Decoction in the treatment of cervical spondylotic myelopathy may be related to the up-regulation of miR-126a-5p expression and the down-regulation of VEGF expression.
3.Development and application of in vitro intestinal absorption model based on microfluidic chips
Lan CHEN ; Xiaoli HE ; Piaoxue YOU ; Hui WANG ; Zhanying HONG
Journal of Pharmaceutical Practice and Service 2024;42(2):43-49
The intestine is the main site of oral drug absorption, and the epithelial cells of the intestine contain villi and microvilli, which promote secretion, cell adhesion, and absorption by increasing surface area and other factors. Traditional two-dimensional/three-dimensional (2D/3D) cell culture models and animal models have played an important role in studying drug absorption, but their application is limited due to the lack of sufficient predictability of human pharmacokinetics or ethical issues, etc. Therefore, mimicking the core structure and key functions of the human intestine based on in vitro live cells has been the focus of research on constructing a microfluidic chip-based intestinal model. The model is a microfluidic chip bionic system that simulates the complex microstructure, microenvironment, and physiological functions of the human intestine using microfabrication technology. Compared with 2D cell culture and animal experiments, the intestinal microarray model can effectively simulate the human in vivo environment and is more specific in drug screening. The research progress and applications in disease modeling, drug absorption and transport of intestinal microarray models and intestine-related multi-organ coupled microarray models at home and abroad were reviewed in this paper. The current challenges of intestinal chip simulating intestinal homeostasis and diseases were summarized,in order to provide reference for the further establishment of a more reliable in vitro intestinal chip model.
4.Sleep quality and influencing factors of mariners during ocean voyages
Zhanying SUN ; Xueyang GAN ; Qingqing SU ; Jingru CHEN ; Yuan GAO
Journal of Environmental and Occupational Medicine 2024;41(8):892-897
Background The average sleep quality of mariners during ocean voyages is notably worse than that of the general populace, and the incidence of sleep disorders among them is higher. Sleep disorders closely associate with fatigue and cognitive decline, increasing error and accident rates, and are a major safety hazard in marine navigation. At present, research on factors influencing the sleep quality of mariners during ocean voyages in China is limited and needs further investigation. Objective To investigate the sleep quality of mariners during ocean voyages and analyze its influencing factors, in order to provide reference for constructing sleep intervention plans and mitigating their sleep disorders. Methods Using convenience cluster sampling, a questionnaire survey was carried out in 408 crew members of a fleet who returned from a voyage on March 21, 2024. The questionnaires included a general information questionnaire, Pittsburgh Sleep Quality Index (PSQI) , and Self Rating Anxiety Scale (SAS), and the data were analyzed by SPSS 26.0 software. Results A total of 399 valid questionnaires were collected, with an effective recovery rate of 97.8%. The mean score of PSQI for the mariners during ocean voyages was (6.41±2.44), with 33.6% (134/399) of the mariners reporting sleep disorders. The PSQI scores varied by family structures (t=2.235, P=0.031), labor types (F=3.789, P=0.023), noise exposure (F=53.218, P<0.001), dietary patterns (F=63.311, P<0.001), exercise habits (F=16.416, P<0.001), and anxiety states (t=5.963,P<0.001). The results of linear regression showed that incomplete family structure (β=0.102, P=0.010), noise exposure (β=0.323, P<0.001), and anxiety (β=0.117, P=0.006) positively associated with the total score of PSQI, while dietary patterns (β=-0.331, P<0.001) and exercise habits (β=-0.147, P<0.001) negatively associated with the total PSQI score, and the 5 variables jointly explained 38.9% of the total variation in the PSQI score (F=37.159, P<0.01). Conclusion The sleep quality of mariners during ocean voyages is relatively low and the incidence of sleep disorders is relatively high, which is jointly influenced by factors such as family structure, noise exposure, dietary habits, exercise habits, and anxiety.
5.Construction of an ultra-early limb movement program for elderly hip fracture surgery patients
Jingru CHEN ; Zhanying SUN ; Xueyang GAN ; Mi SONG ; Jie SONG ; Yuan GAO
Chinese Journal of Modern Nursing 2024;30(34):4688-4695
Objective:To construct an ultra-early limb movement program for elderly hip fracture surgery patients based on evidence-based practice and the Delphi method.Methods:From May to December 2023, the PIPOST Model (population, intervention, professional, outcome, setting, type of evidence) was used to develop evidence-based questions. Relevant databases were searched to obtain, summarize, and evaluate evidence related to ultra-early limb movement after hip fracture surgery in elderly patients. A draft of the movement program was created with the search timeline set until June 1, 2023. A total of 29 experts were invited to participate in two rounds of Delphi expert consultation to finalize the ultra-early limb movement program for elderly hip fracture surgery patients.Results:A total of 20 studies were included, consisting of six guidelines, four expert consensuses, three systematic reviews, and seven original research articles. The response rates for the two rounds of Delphi expert consultation were 100.00% (29/29) and 86.21% (25/29). The expert authority coefficients were 0.931 and 0.920, and the Kendall coordination coefficients were 0.205 and 0.112 (all P<0.01). The final ultra-early limb movement program for elderly hip fracture surgery patients included four modules and 39 operational items. Conclusions:This study develops an evidence-based, expert-validated ultra-early limb movement program for elderly hip fracture surgery patients, demonstrating its necessity, scientific basis, and practicality. The program provides a reference and guidance for clinical staff in facilitating ultra-early movement for hip fracture patients post-surgery.
6.Association Between Normal-weight Central Obesity With New-onset Cardiovascular Disease and All-cause Mortality
Zhanying MA ; Jierui WANG ; Haicheng SONG ; Fan YANG ; Jiaoyan LI ; Mingzhu ZHAO ; Lizhi CHEN ; Lina LI ; Wenfang YANG ; Shuohua CHEN ; Shouling WU ; Liufu CUI
Chinese Circulation Journal 2024;39(11):1110-1116
Objectives:To investigate the association between normal-weight central obesity with new-onset cardiovascular disease and all-cause mortality risk. Methods:A prospective cohort study was conducted,selecting a total of 93885 participants from the Kailuan Study who had their first physical examination in 2006-2007.According to waist circumference (central obesity:male waist circumference ≥90 cm,female waist circumference ≥85 cm;no central obesity:male waist circumference<90 cm,female waist circumference<85 cm) and body mass index (BMI,normal weight:18.5 kg/m2≤BMI<24.0 kg/m2;overweight/obesity:BMI ≥24.0 kg/m2),the participants were divided into 4 groups:normal weight no central obesity group (G1 group),normal weight central obesity group (G2 group),overweight/obesity no central obesity group (G3 group) and overweight/central obesity group (G4 group);Using the Kaplan-Meier method,the cumulative incidence of new-onset cardiovascular diseases (including hemorrhagic stroke,ischemic stroke and myocardial infarction) and all-cause mortality in different groups was calculated,and the Log-rank test was used for intergroup comparisons.Furthermore,the associations between the different groups and the risk of new-onset cardiovascular diseases and all-cause mortality were analyzed using the multivariate Cox proportional hazard regression model. Results:After a median follow-up of 14.97 (14.55,15.17) years,the cumulative incidence of new-onset cardiovascular diseases in G1 group,G2 group,G3 group and G4 group was 7.62%,10.84%,8.67%,12.91% respectively (log-rank P<0.05) and the cumulative incidence of all-cause mortality was 12.83%,19.72%,10.65%,16.33% respectively (log-rank P<0.01).After adjusting for confounding factors,Cox regression analysis showed that the HR (95%CI) of new-onset cardiovascular diseases in G2 group,G3 group and G4 group were 1.14 (1.04-1.25),1.07 (1.01-1.14),1.27 (1.21-1.34),respectively compared with G1 group (all P<0.05).The HR (95%CI) of all-cause mortality were 1.06 (1.00-1.14),0.90 (0.85-0.95),0.97 (0.93-1.01) compared with G1 group,and P values were 0.07,<0.01,0.15,respectively.The results of sensitivity analysis were consistent with the above major studies after excluding overweight/obesity and cancer participants during follow-up. Conclusions:Normal-weight central obesity increases the risk of new-onset cardiovascular diseases and all-cause mortality.
7.Expert consensus on the accurate diagnosis and treatment of acetabular fractures based on three-column classification (version 2023)
Ruipeng ZHANG ; Hongmin CAI ; Shicai FAN ; Gang LYU ; Yan ZHUANG ; Chengla YI ; Xiaodong GUO ; Longpo ZHENG ; Xianzhong MA ; Hua CHEN ; Dahui SUN ; Guanglin WANG ; Qishi ZHOU ; Weixu LI ; Wei FENG ; Zhangyuan LIN ; Xiaodong QIN ; Jiandong WANG ; Zhanying SHI ; Lianxin LI ; Guangyao LIU ; Shuquan GUO ; Ming LI ; Jianzhong GUAN ; Yingze ZHANG ; Zhiyong HOU
Chinese Journal of Trauma 2023;39(10):865-875
Accurate classification of the acetabular injuries and appropriate treatment plan are great challenges for orthopedic surgeons because of the irregular anatomical structure of the acetabulum and aggregation of important vessels and nerves around it. Letournel-Judet classification system has been widely applied to classify acetabular fractures. However, there are several limitations, including incomplete inclusion of fracture types, difficulty in understanding and insufficient guidance for surgical treatment, etc. Serious complications such as traumatic arthritis are common due to wrong classification and diagnosis and improper selection of surgical strategy, which brings a heavy burden to the society and families. Three-column classification, based on anatomic characteristics, has advantages of containing more fracture types and being easy to understand, etc. To solve the problems existing in the diagnosis and treatment process based on Letournel-Judet classification, achieve accurate diagnosis and treatment of patients with acetabular fractures, and obtain satisfactory prognosis, the Orthopedic Trauma Emergency Center of Third Hospital of Hebei Medical University and the Trauma Orthopedic Branch of the Chinese Orthopedic Association organized experts from relevant fields to formulate the Expert consensus on the accurate diagnosis and treatment of acetabular fractures based on three-column classification ( version 2023) in terms of principles of evidence-based medicine. Based on the three-column classification, 15 recommendations were proposed, covering the diagnosis, treatment, complication prevention and management, etc, so as to provide reference for accurate diagnosis and treatment of acetabular fractures.
8.Evaluation of toxin-induced injury and protective effect of triptolide based on a cardiovascular chip model
Xiaoli HE ; Yiwei SHI ; Lan CHEN ; Yue LIU ; Zhanying HONG
Journal of Pharmaceutical Practice 2023;41(3):149-154
Objective To construct a cardiovascular chip model for evaluating the damage of vascular glycocalyx induced by four marine toxins: okadaic acid (OA), conotoxin (CTX), tetrodotoxin (TTX) and gymnodimine (GYM), and explore the protective effect of triptolide on toxin-induced injury. Methods Human umbilical vein endothelial cells(HUVEC) were inoculated into a three-channel microfluidic chip. CCK-8 method and immunofluorescence staining were used to analyze the damage of cell viability and glycocalyx tissue induced by low, middle and high concentrations of marine toxin, as well as the protective effect of triptolide on toxin-induced injury. Results The cells in the cardiovascular chip grew well and had structurally intact glycocalyx. Compared with the control group, the activity of HUVEC cells were inhibited in group of the medium and high concentration of OA and high concentration of GYM (P<0.05). The activity of cells had not been inhibited by CTX and TTX significantly , but all the four toxins caused serious damage to the glycocalyx tissue (P<0.01). After pre-protection with triptolide, the toxicity of the four toxins to HUVEC cells and the damage rate of glycocalyx decreased significantly. Conclusion The four marine biotoxins could damage the activity and glycocalyx of HUVEC cells in a dose-dependent manner, while triptolide has a protective effect on HUVEC cells injured by toxin.
9.Screening of immune cell activators from Astragali Radix using a comprehensive two-dimensional NK-92MI cell membrane chromatography/C18 column/time-of-flight mass spectrometry system
Xinyi CHAI ; Yanqiu GU ; Lei LV ; Chun CHEN ; Fei FENG ; Yan CAO ; Yue LIU ; Zhenyu ZHU ; Zhanying HONG ; Yifeng CHAI ; Xiaofei CHEN
Journal of Pharmaceutical Analysis 2022;12(5):725-732
Astragali Radix(AR)is a clinically used herbal medicine with multiple immunomodulatory activities that can strengthen the activity and cytotoxicity of natural killer(NK)cells.However,owing to the complexity of its composition,the specific active ingredients in AR that act on NK cells are not clear yet.Cell membrane chromatography(CMC)is mainly used to screen the active ingredients in a complex system of herbal medicines.In this study,a new comprehensive two-dimensional(2D)NK-92MI CMC/C18 column/time-of-flight mass spectrometry(TOFMS)system was established to screen for potential NK cell acti-vators.To obtain a higher column efficiency,3-mercaptopropyltrimethoxysilane-modified silica was synthesized to prepare the NK-92MI CMC column.In total,nine components in AR were screened from this system,which could be washed out from the NK-92MI/CMC column after 10 min,and they showed good affinity for NK-92MI/CMC column.Two representative active compounds of AR,isoastragaloside Ⅰ and astragaloside Ⅳ,promoted the killing effect of NK cells on K562 cells in a dose-dependent manner.It can thus suggest that isoastragaloside Ⅰ and astragaloside Ⅳ are the main immunomodulatory compo-nents of AR.This comprehensive 2D NK-92MI CMC analytical system is a practical method for screening immune cell activators from other herbal medicines with immunomodulatory effects.
10.Development of a surface plasmon resonance biosensor for accurate and sensitive quantitation of small molecules in blood samples
Minyu QI ; Diya LV ; Ying ZHANG ; Dongyao WANG ; Xiaofei CHEN ; Zhenyu ZHU ; Zhanying HONG ; Yifeng CHAI ; Hai ZHANG ; Yan CAO
Journal of Pharmaceutical Analysis 2022;12(6):929-936
Therapeutic drug monitoring(TDM)has played an important role in clinical medicine for precise dosing.Currently,chromatographic technology and immunoassay detection are widely used in TDM and have met most of the needs of clinical drug therapy.However,some problems still exist in practical appli-cations,such as complicated operation and the influence of endogenous substances.Surface plasmon resonance(SPR)has been applied to detect the concentrations of small molecules,including pesticide residues in crops and antibiotics in milk,which indicates its potential for in vivo drug detection.In this study,a new SPR-based biosensor for detecting chloramphenicol(CAP)in blood samples was developed and validated using methodological verification,including precision,accuracy,matrix effect,and extraction recovery rate,and compared with the classic ultra-performance liquid chromatography-ultraviolet(UPLC-UV)method.The detection range of SPR was 0.1-50 ng/mL and the limit of detec-tion was 0.099±0.023 ng/mL,which was lower than that of UPLC-UV.The intra-day and inter-day ac-curacies of SPR were 98%-114%and 110%-122%,which met the analysis requirement.The results show that the SPR biosensor is identical to UPLC-UV in the detection of CAP in rat blood samples;moreover,the SPR biosensor has better sensitivity.Therefore,the present study shows that SPR technology can be used for the detection of small molecules in the blood samples and has the potential to become a method for therapeutic drug monitoring.

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