1.Effect of Wenshen Tongluo Zhitong formula on mouse H-type bone microvascular endothelial cell/bone marrow mesenchymal stem cell co-culture system
Shijie ZHOU ; Muzhe LI ; Li YUN ; Tianchi ZHANG ; Yuanyuan NIU ; Yihua ZHU ; Qinfeng ZHOU ; Yang GUO ; Yong MA ; Lining WANG
Chinese Journal of Tissue Engineering Research 2025;29(1):8-15
BACKGROUND:Bone relies on the close connection between blood vessels and bone cells to maintain its integrity.Bones are in a physiologically hypoxic environment.Therefore,the study of angiogenesis and osteogenesis in hypoxic environment is closer to the microenvironment in vivo. OBJECTIVE:To explore the influence of Wenshen Tongluo Zhitong(WSTLZT)formula on H-type bone microvascular endothelial cell/bone marrow mesenchymal stem cell co-culture system in hypoxia environment and its related mechanism. METHODS:Enzyme digestion method and flow sorting technique were used to isolate and identify H-type bone microvascular endothelial cells.Mouse bone marrow mesenchymal stem cells were isolated and obtained by bone marrow adhesion method.H-type bone microvascular endothelial cell/bone marrow mesenchymal stem cell hypoxic co-culture system was established using Transwell chamber and anoxic culture workstation.WSTLZT formula powder was used to intervene in each group at a mass concentration of 50 and 100 μg/mL.The angiogenic function of H-type bone microvascular endothelial cells in the co-culture system was evaluated by scratch migration test and tube formation test.The osteogenic differentiation ability of bone marrow mesenchymal stem cells in the co-cultured system was evaluated by alkaline phosphatase staining and alizarin red staining.The protein and mRNA expression changes of PDGF/PI3K/AKT signal axis related molecules in H-type bone microvascular endothelial cells in the co-cultured system were detected by Western Blotting and q-PCR,respectively. RESULTS AND CONCLUSION:(1)Compared with the normal oxygen group,the scratch mobility and new blood vessel length of H-type bone microvascular endothelial cells were significantly higher(P<0.05);the osteogenic differentiation capacity of bone marrow mesenchymal stem cells was higher(P<0.05);the expression of PDGF/PI3K/AKT axis-related molecular protein and mRNA increased(P<0.05)in the hypoxia group.(2)Compared with the hypoxia group,scratch mobility and new blood vessel length were significantly increased in the H-type bone microvascular endothelial cells(P<0.05);bone marrow mesenchymal stem cells had stronger osteogenic function(P<0.05);the expression of PDGF/PI3K/AKT axis-related molecular proteins and mRNA further increased(P<0.05)after treatment with different dose concentrations of WSTLZT formula.These findings conclude that H-type angiogenesis and osteogenesis under hypoxia may be related to the PDGF/PI3K/AKT signaling axis,and WSTLZT formula may promote H-type vasculo-dependent bone formation by activating the PDGF/PI3K/AKT signaling axis,thereby preventing and treating osteoporosis.
2.Investigation and Analysis of the Fourth National General Investigation on the Medicinal Plant Resources in Hongya County of Sichuan Province
Lei ZHAO ; Yuntan QIN ; Shijie ZHU ; Qingmao FANG ; Bing LUO ; Xianjian ZHOU ; Jnuning ZHAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(8):1967-1974
Objective To investigate the medicinal plant resources in Hongya County of Sichuan Province,so as to provide data support for the development,utilization,and protection of traditional Chinese medicine resources in Hongya County.Methods This study conducted a comprehensive investigation on the medicinal plant species in Hongya County using literature review,field investigation,interview investigation,sample plot investigation,and specimen collection.Results The findings revealed a total of 1748 species of medicinal plants in Hongya County,belonging to 808 genera and 185 families,the Compositae family had the highest proportion.Furthermore,the study identified 5 rare and endangered medicinal plants,14 species of wild plants under state key protection,9 species of wild medicinal materials under state key protection,185 species of traditional Chinese medicine listed in the 2020 edition of Chinese Pharmacopoeia of the People's Republic.Conclusion The medicinal plant resources in Hongya County are rich,but the utilization rate is low,and there is a large space for the industrialization development of traditional Chinese medicine.The general investigation results provide a reference for the rational development and protection of medicinal plant in Hongya County.
3.Precision nutritional therapy in gastrointestinal tumor
Da ZHOU ; Shijie WANG ; Xinying WANG
Chinese Journal of Gastrointestinal Surgery 2024;27(3):225-230
Apart from individual genetic background, unhealthy lifestyle and diet, etc., nutrition also plays an important role in the occurrence and progression of gastrointestinal tumors. Although some patients with gastrointestinal tumors can be satisfied with the traditional nutritional support, it is apparently inadequate for the systemic management of all patients. Precision nutrition support, also known as personalized nutrition support, refers to safe and efficient individualized nutrition intervention based on the investigation of individual genetic background, life characteristics, metabolic indicators, intestinal microbial characteristics, and physiological status factors through big data analysis for the prevention and treatment of chronic diseases. This review focuses on the relationship between nutrition and gastrointestinal tumors and discusses the progress of precision nutrition support therapy in the gastrointestinal tumors. Based on this, we hope to achieve effective personalized intervention protocols, and improve the clinical outcome and the overall oncology care of gastrointestinal tumors.
4.Levels of sICAM-1 and sVCAM-1 and activity of SOD in serum and their relationships with coronary artery calcification in patients with maintenance hemodialysis
Yuwei KANG ; Wei YANG ; Shijie MA ; Wei ZHOU ; Fei DENG
Journal of Jilin University(Medicine Edition) 2024;50(3):812-818
Objective:To analyze the levels of serum soluble intercellular adhesion molecule-1(sICAM-1),soluble vascular cell adhesion molecule-1(sVCAM-1),and superoxide dismutase(SOD)activity in the patients with maintenance hemodialysis(MHD),and to discuss their relationships with coronary artery calcification(CAC).Methods:The clinical materials from 102 MHD patients(MHD group)were retrospectively analyzed.Additionally,74 volunteers underwent routine health examination at the same time(health examination group)were selected.The CAC scores(CACs)of the patients in MHD group were detected by multi-slice computed tomography(MSCT),and the patients were categorized into non-calcification group,mild calcification group,moderate calcification group,and severe calcification group.The general data and serum levels of sICAM-1,sVCAM-1,and SOD activities of the subjects in two groups were compared.The levels of calcium(Ca),phosphorus(P),parathyroid hormone(PTH),sICAM-1,sVCAM-1,and SOD activities in serum of the patients with different degrees of calcification were analyzed.Pearson's correlation analysis was used to analyze the correlations between the levels of sICAM-1,sVCAM-1,and SOD activity in serum of the MHD patients and CACs.Results:Compared with health examination group,the levels of sICAM-1 and sVCAM-1 in serum of the patients in MHD group were significantly increased(P<0.01),and the SOD activity was significantly decreased(P<0.01).Compared with non-calcification group,the levels of PTH,sICAM-1,and sVCAM-1 in serum of the MHD patients in mild,moderate,and severe calcification groups were significantly increased(P<0.05),and the SOD activities were significantly decreased(P<0.05);the levels of P in serum of the MHD patients in moderate and severe calcification groups were significantly increased(P<0.05).Compared with mild calcification group,the levels of P,PTH,sICAM-1,and sVCAM-1 in serum of the MHD patients in moderate and severe calcification groups were significantly increased(P<0.05),and the SOD activities significantly decreased(P<0.05).Compared with moderate calcification group,the levels of sICAM-1,sVCAM-1,and P in serum of the MHD patients in severe calcification group were significantly increased(P<0.05),and the SOD activity was significantly decreased(P<0.05).The SOD activity in serum of the MHD patients was negatively correlated with CACs(r=-0.484,P<0.01),while the levels of sICAM-1 and sVCAM-1 were positively correlated with CACs(r=0.441,P<0.01;r=0.561,P<0.01).Conclusion:The levels of sICAM-1 and sVCAM-1 and activity of SOD in serum of the MHD patients are abnormal.With the decreasing of the SOD activity and increasing of the levels of sICAM-1 and sVCAM-1,the degree of CAC in the MHD patients is aggravated.
5.Analysis of the three-dimensional image characteristics of bone island in jaw using CBCT
Shijie YAN ; Lianfeng YANG ; Jingyan WANG ; Juan ZHANG ; Tingting ZHU ; Diya LENG ; Guangchao ZHOU ; Peiyu GU
STOMATOLOGY 2024;44(10):721-727
Objective To analyze three-dimensional(3D)radiographic characterizations of bone island(BI)in jaw using cone-beam computed tomography(CBCT).Methods CBCT data from four thousand patients were selected,reconstructed and analyzed using NNT 10.0 software.The sagittal,coronal and axial planes were used to analyze the 3D radiographic characteristics of BIs,including the localization,shape,density,boundary,the relationship between BIs and tooth and bone cortex,diameter and anatomical structures and complications involved.Their relationship with gender were analyzed.Results A total of 803 people had BIs,with the prevalence rate of 20.08%,including 338 males with 389 BIs and 465 females with 526 BIs.Both males and females had a dominant BI,and the ratio between male and female was 1∶1.38,but the difference was not statistically significant(P>0.05).The BIs of both male and female mostly occurred in the mandibular premolars and molars area,and appeared irregular in shape,dense and contact with lingual bone cortex.Mostly BIs were apical type and with unclear boundary.The mean maximum diameter of mesial/distal direction was greater than buccal/lingual direction(P<0.05).The most commonly involved anatomy structure was the inferior alveolar neural canal,cortical infil-tration and mental foramen.Conclusion There are no significant differences between males and females in the three-dimensional image characteristics of BIs in Chinese populations.CBCT can accurately and comprehensively analyze the 3D radiographic characteris-tics of BI and its relationship with the surrounding teeth and bone.
6.POLG inhibitor suppresses migration and invasion of triple-negative breast cancer cells via blocking mitochondrial biogenesis
Xing LIU ; Shuangqin FAN ; Xiaomin YAN ; Shijie ZHAO ; Rong WANG ; Xiangchun SHEN ; Xue ZHOU ; Yue ZHANG ; Yan CHEN
Acta Universitatis Medicinalis Anhui 2024;59(10):1720-1728
Objective To investigate the effects of zalcitabine(ddC),a mitochondrial DNA polymerase γ(POLG)inhibitor,on the migration,invasion,and to preliminarily explore mitochondrial biogenesis of human tri-ple-negative breast cancer MDA-MB-231 cells.Methods The effect of ddC on cell viability was detected using the MTT assay.The migration and invasion abilities of the cells were evaluated using the cell scratch and Transwell in-vasion assays.Cell apoptosis was determined using flow cytometry and a V-FITC/PI cell apoptosis detection kit.The protein expression of POLG,NADH dehydrogenase subunit Ⅰ(NADH1),NADH dehydrogenase subunit Ⅱ(NADH2),ATP synthase subunit 6(ATPase6),cytochrome c oxidase subunit Ⅰ(COX-1)and cytochrome c ox-idase subunit Ⅲ(COX-3)were determined using Western blot.The POLG mRNA level and mtDNA copy number were determined using qPCR.The mitochondrial content and ATP levels were determined using MitoTracker Green fluorescent probe staining and an ATP determination kit.MDA-MB-231 cells were transfected with pcDNA3.1-EG-FP-POLG plasmids to overexpress POLG.The inhibitory effects of ddC on cell migration and invasion were detected in POLG-overexpressed MDA-MB-231 cells.Results POLG expression was higher in MDA-MB-231 cells than in normal mammary epithelial cells(MCF-10A)(P<0.01).ddC inhibited cell viability in a dose-dependent man-ner.ddC inhibited the migration(P<0.01)and invasion(P<0.01)of MDA-MB-231 cells;however,it dis-played no significant inhibitory effects on cell viability in normal mammary epithelial cells(MCF-10A)at the same concentration.ddC downregulated the protein(P<0.01)and mRNA(P<0.01)levels of POLG,reduced mtD-NA copy number(P<0.01)and downregulated mtDNA-coded NADH1,NADH2,ATPase6,COX-1 and COX-3 protein expression(P<0.01)in MDA-MB-231 cells.Furthermore ddC inhibited mitochondrial content(P<0.01)and ATP(P<0.01)levels in MDA-MB-231 cells.POLG overexpression increased the migration(P<0.05)and invasion(P<0.05)abilities of MDA-MB-231 cells,while ddC did not significantly inhibit the migra-tion and invasion abilities of MDA-MB-231 cells overexpressing POLG.Conclusion ddC downregulates POLG ex-pression in MDA-MB-231 cells and inhibits mitochondrial biogenesis and ATP levels,thereby inhibiting the migra-tion and invasion of MDA-MB-231 cells.
7.Mugwort pollen-induced mouse allergic asthma and endotyping
Linghui ZHOU ; Linmei LI ; Huancheng XIE ; Shijie SONG ; Ying HE ; Ailin TAO
Chinese Journal of Immunology 2024;40(1):52-57
Objective:To construct a mouse asthma model induced by mugwort pollen and to explore endotyping,providing methods for subsequent precision treatment.Methods:BALB/c mice were intraperitoneally injected with mugwort pollen extract(MPE)to sensitize,following MPE intranasal challenge to construct MPE allergic asthma murine model.Mice were randomly divided into PBS sensitization and PBS challenge(P-P),MPE sensitization and PBS challenge(M-P),MPE sensitization and MPE challenge model(M-M)groups.24 h after final challenge,mice were performed to examine airway responsiveness;bronchoalveolar lavage fluid(BALF)was harvested for cell counting and statistical classification of inflammatory cells through flow cytometry analysis.Pulmonary slides were collected for pathological examination,including HE,PAS,Masson and α-SMA immunohistochemical staining.ELISA was used to detect levels of IFN-γ,IL-4,IL-5,IL-13,IL-17A in lung tissue and serum,as well as serum total IgE and MPE-specific IgE,IgG1,IgG2a levels.Results:Pathological examination showed higher airway reactivity,more inflammatory cells infiltration around airway,obvious goblet metaplasia,thickening of airway smooth muscles and dramatical fibrin deposition around airway in model group.Total cell numbers of BALF were increased from<1×105 cells/ml in P-P group to>5×105 cells/ml in model group,in which eosinophils were predominant cellular type,levels of IL-4,IL-13,IL-17A in lung and IL-5,IL-13 levels in serum were significantly increased,as well as significant increasing levels of total IgE and MPE-specific IgE,IgG1,IgG2a.Conclusion:MPE-sensitized and challenged mice induces typical eosinophilic asthma featured with elevated eosinophils,as well as secretion of inflammatory factors of type 2 and type 17,IgE,IgG1 and IgG2a subtypes soars to high levels.
8.Effects of three rehydration methods on prevention of on-site and delayed blood donation-related vasovagal responses: a cluster-randomized trial
Guiyun XIE ; Shijie LI ; Jian OUYANG ; Fanfan FENG ; Xiaoxiao ZHENG ; Zhiyu ZHOU ; Lianfang MAI ; Jinyan CHEN
Chinese Journal of Blood Transfusion 2024;37(1):43-50
【Objective】 To compare the effects of 3 rehydration methods before blood donation on the prevention of on-site and delayed blood donation-related vasovagal response (VVR) . 【Methods】 From January to June 2021, 6 250 whole blood donors in 6 fixed blood donation sites signed informed consent and were divided into 198 clusters according to donor sites and dates, then they were randomly assigned to receive either oral rehydration salts (ORS), sugar water, or water group, and each drank 500 mL of ORS, sugar water or water within 20 minutes before blood donation. The researchers recorded the actual intervention accepted on site, and recorded the immediate VVR and related information. At rest after blood donation, donors submitted an electronic questionnaire containing socio-demographic information. At 48 hours after blood donation, the researchers called back every donor to record delayed VVR and related information. Logistic regression based on intention to treat (ITT) was used to analyze the difference of the incidence of VVR among the three groups, and the average treatment effect on treated (ATT) was calculated. PASS 2021was used to estimate the sample size and R (4.2.0) for statistical analysis. 【Results】 The cumulative incidence of blood donation-related VVR was 2.67% (2.29%-3.11%) among street whole blood donors under the 3 rehydration methods, in which, the incidence of immediate and delayed VVR was 1.02% (0.79%-1.31%) and 1.65% (1.36%-2.01%) respectively. ITT analysis found that ORS were more effective than water in reducing the incidence of delayed VVR【OR=0.59,95% CI[0.37,0.94]】.There was no significant difference in the incidence of immediate VVR between any two groups (P > 0.05), and there was no significant difference in the incidence of delayed VVR in the sugar water group compared with the water group (P > 0.05). There was a difference of -0.013 (【95% CI[-0.022, -0.004]】or -0.008【95% CI[-0.017, -0.000]】in the incidence of delayed VVR in the ORS group compared with water group or sugar water group, the difference was significant (P<0.05). The cumulative VVR of the three groups showed similar results to the delayed VVR. 【Conclusion】 Drinking ORS before blood donation is the most effective rehydration method to prevent delayed VVR. The next step is to establish the predictive model of delayed VVR to screen the susceptible population and provide them with ORS before blood donation, while other population can choose any liquid they like, thus achieving personalized blood donation-related VVR prevention and control.
9.Model of "One Core, Multiple Elements" emergency blood donation volunteer team: a case study of Nanjing city
Shangyun YING ; Yudong DAI ; Yilun ZHAO ; Shijie ZHOU ; Huifang ZHOU
Chinese Journal of Blood Transfusion 2024;37(1):84-90
【Objective】 To explore the model of "One Core, Multiple Elements" emergency blood donation volunteer team (referred to as the Model) . 【Methods】 The Nanjing City Voluntary Blood Donation Joint Meeting serves as the core, with diverse entities including party committees, government departments, district governments, social organizations, enterprises, blood donors, etc. Following the principles of "emergency response in emergencies, wartime readiness, combining regular and wartime efforts," and adhering to the framework of the Model, the emergency blood donation volunteer team system in Nanjing was constructed. 【Results】 1) After the construction of the Model (2018—2022), the total number of emergency blood donation volunteers in Nanjing City increased by 191% compared to the pre-construction period (2013—2017), with an average annual blood donation of 20 929, showing significant differences (P<0.05). 2) After the construction of the Model, the number of emergency blood donation during the winter and summer increased by 206% and 185%, respectively, compared to the pre-construction period, demonstrating significant differences (P< 0.05) .3) Prior to the construction of the Model, Nanjing lacked a relatively stable emergency volunteer team. After the construction, Nanjing established a total of 5 relatively stable volunteer teams, with the district-level government and township personnel accounting for the highest proportion (52.96%), followed by medical personnel (23.95%), enterprise team (11.10%), State-owned Assets Supervision and Administration Commission team (7%), and Nanjing municipal government team (4.98%). 4) Following the Model, during the COVID-19 pandemic, Nanjing successfully initiated two Level Ⅱ blood emergency responses, with emergency blood donations reaching 23 041. 【Conclusion】 The Model can effectively ensure the blood supply in Nanjing region during emergencies.
10.Molecular typing and drug resistance of Salmonella in market-sold raw livestock meats in Shaoxing City
HE Qinfen ; JIANG Zhuojing ; TANG Shijie ; ZHOU Liangkang ; ZHANG Qinchao
Journal of Preventive Medicine 2024;36(10):915-920
Objective:
To investigate the molecular typing and drug resistance of Salmonella in market-sold raw livestock meats in Shaoxing City, Zhejiang Province, so as to provide the basis for the prevention and control of drug-resistant Salmonella.
Methods:
Salmonella isolates were detected and serotyped from raw livestock meat samples collected from supermarkets, agricultural markets and restaurants in Shaoxing City between March and November 2023. Molecular typing was performed using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Antibiotic resistance phenotypes and resistance gene analyses were conducted using broth microdilution method and whole genome sequencing.
Results:
A total of 34 Salmonella isolates were detected from 329 samples and the positive rate was 10.33%. There were 14 serotypes, with S. Rissen (9 isolates, 26.47%) and S. London (8 isolates, 23.53%) being dominant. PFGE typing revealed 31 band patterns and 23 banding pattern clusters of Salmonella, and the similarity ranged from 30.80% to 100.00%, without dominant band patterns. MLST identified 15 sequence typing (ST) types, and S. Rissen were all ST469 and S. London were all ST155. Salmonella strains exhibited high levels of resistance to tetracycline (44.12%) and ampicillin (35.29%). Thirteen resistance patterns were identified, with 8 multidrug resistant patterns (29.41%). There were 8 classes of 14 resistance genes, with aminoglycoside resistance gene aac(6' )-Iaa found in 100.00% of the strains. The highest concordance rate between the resistance phenotype and gene was observed for phenicol antibiotics (97.06%), while there was no concordance between the quinolone resistance phenotype and genes.
Conclusions
S. Rissen and S. London are the predominant serotypes in market-sold raw livestock meats in Shaoxing City. The PFGE band types of the Salmonella isolates show polymorphism distribution, and MLST are mainly ST469 and ST155. The isolates have high levels of resistance to tetracycline and ampicillin, manifesting multidrug resistance.


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