1.Single-Cell RNA Sequencing of Bone Marrow Mesenchymal Stem Cells from the Elderly People
Dezhou ZHU ; Jie GAO ; Chengxuan TANG ; Zheng XU ; Tiansheng SUN
International Journal of Stem Cells 2022;15(2):173-182
Background and Objectives:
Bone marrow mesenchymal stem cells (BMSCs) show considerable promise in regenerative medicine. Many studies demonstrated that BMSCs cultured in vitro were highly heterogeneous and composed of diverse cell subpopulations, which may be the basis of their multiple biological characteristics. However, the exact cell subpopulations that make up BMSCs are still unknown.
Methods:
and Results: In this study, we used single-cell RNA sequencing (scRNA-Seq) to divide 6,514 BMSCs into three clusters. The number and corresponding proportion of cells in clusters 1 to 3 were 3,766 (57.81%), 1,720 (26.40%), and 1,028 (15.78%). The gene expression profile and function of the cells in the same cluster were similar. The vast majority of cells expressed the markers defining BMSCs by flow cytometry and gene expression analysis. Each cluster had at least 20 differentially expressed genes (DEGs). We conducted Gene Ontology enrichment analysis on the top 20 DEGs of each cluster and found that the three clusters had different functions, which were related to self-renewal, multilineage differentiation and cytokine secretion, respectively. In addition, the function of the top 20 DEGs of each cluster was checked by the National Center for Biotechnology Information gene database to further verify our hypothesis.
Conclusions
This study indicated that scRNA-Seq can be used to divide BMSCs into different subpopulations, demon-strating the heterogeneity of BMSCs.
2.Single cell RNA sequencing of bone marrow mononuclear cells from elderly patients with hip fracture
Dezhou ZHU ; Jie GAO ; Zhijie ZHENG ; Yun LAN ; Zheng XU ; Jianzheng ZHANG ; Zhi LIU ; Tiansheng SUN
Chinese Journal of Orthopaedic Trauma 2021;23(6):519-525
Objective:To determine whether subpopulations may exist which are related to regulation of immunization or inflammation in bone marrow mononuclear cells (BMMNCs) from 2 elderly patients with hip fracture and whether there might be any difference in the subpopulations between them.Methods:Two elderly patients with hip fracture were enrolled in this study. Their venous blood was harvested to determine subpopulations of complement (C)3, C4, interleukins (IL)-2, IL-6, IL-10, and lymphocytes. Single cell RNA sequencing (scRNA-Seq) was used to group their BMMNCs. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed of the top 20 differentially expressed genes for each subpopulation to judge the main function of each subpopulation. The subpopulations and the key genes related to regulation of immunization or inflammation were found out. The relationships were explored between subpopulations and prognosis of the patients.Results:By the venous blood indexes, IL-10 was slightly high in patient A; C3 below normal, C4 close to the lower normal limit, IL-2, IL-6 and IL-10 were significantly high, CD8 +T % was low, and CD4 +/CD8 + high in patient B. After scRNA-Seq and bioinformatics analyses, the BMMNCs in the 2 patients were divided into 5 subpopulations. GO and KEGG enrichment analyses showed that the functions of subpopulation 2 and subpopulation 4 were related to immunization or inflammation. CCL4, CCL5, LTB and CXCR4 in subpopulation 2 and C1QA, C1QB, CD14 and SPP1 in subpopulation 4 were related to the regulation of immunization or inflammation. The final prognosis of patient A was much better. The proportions of BMMNCs involved in subpopulation 2 and subpopulation 4 from patient A were higher than those from patient B [47.00% (1,431/3,045) versus 29.28% (882/3,012); 5.88% (179/3,045) versus 3.85% (116/3,012)]. Conclusions:The BMMNCs from elderly patients with hip fracture can be divided into subpopulations by scRNA-Seq. Some of the subpopulations may be related to regulation of immunization or inflammation, which may affect the post-injury immune inflammatory state and prognosis of the patients.
3.Coma in spontaneous intracranial hypotension complicated with subdural hematoma: a case report
Zhen WANG ; Yanfang DAI ; Yanhong AN ; Yanhui MA ; Jie WU ; Dezhou QI ; Jingjing SUN ; Hong YE ; Ran GAO ; Junjie LI ; Liyong WU
Chinese Journal of Neurology 2022;55(4):349-353
Spontaneous intracranial hypotension can frequently result in several complications including subdural hygroma, subdural hematoma and cerebral venous thrombosis, but coma rarely. A case of spontaneous intracranial hypotension presented with orthostatic headaches was described. He experienced somnolence, disorientation, incontinence, and then coma, though received conservative treatment. Brain imaging demonstrated acute-on-chronic subdural hematoma, magnetic resonance myelography using heavily T 2-weighted fast spin-echo pulse sequences showed spinal longitudinal extradural collection, and magnetic resonance myelography with intrathecal gadolinium revealed cerebrospinal fluid leak at the level of T 6, T 7. The patient recovered consciousness after surgical evacuation of the hematoma, and the headache disappeared after a targeted epidural blood patch. The hematoma resolved 2 months later and the patient kept free from headache during follow-up.
4.A case report of flow diverters grafting for the treatment of neurofibromatosis type Ⅰ-related pediatric intracranial aneurysm
Lin SHI ; Bingbo LYU ; Baoxin REN ; Min WANG ; Dezhou SUN ; Donghai WANG
Chinese Journal of Cerebrovascular Diseases 2024;21(7):472-475
Intracranial aneurysms in children are rare in clinical practice.Its pathogenesis is not clear,and the treatment method has many controversies.The authors reported a case of pediatric intracranial aneurysms associated with neurofibromatosis type Ⅰ treated with two flow diverters,and reviewed relevant literatures,hoping to provide a useful reference for exploring the treatment of pediatric intracranial aneurysms.
5.Simultaneous Determination of 5 Components in the Classical Formula Huaihua San by HPLC Wavelength Switching Method
Zedong XIANG ; Qing XUE ; Haitao YU ; Zhiqiang SUN ; Zhen LI ; Juyan LIU ; Jiayu ZHANG ; Peng GAO ; Long DAI
China Pharmacy 2021;32(16):1964-1968
OBJECTIVE:To develop a method for simultaneous determination of 5 components in classical formula Huaihua san,including rutin ,naringin,neohesperidin,quercetin and pulegone. METHODS :HPLC wavelength switching method was adopted. The determination was performed on Cosmosil C 18 column with mobile phase consisted of acetonitrile- 0.05% phosphoric acid solution (gradient elution )at the flow rate of 1.0 mL/min. The detection wavelengths were set at 257 nm for rutin ,283 nm for naringin and neohesperidin ,254 nm for quercetin ,252 nm for pulegone ,respectively. The column temperature was set at 30 ℃, and sample size was 10 μL. RESULTS:The linear range was 21.7-2 170 μg/mL for rutin,46-4 600 μg/mL for naringin,22.3- 2 230 μg/mL for neohesperidin,0.96-96 μg/mL for quercetin,2.7-270 μg/mL for pulegone(all r>0.999),respectively. RSDs of precision,stability(24 h)and reproducibility tests were all lower than 2%(n=6). Average recoveries were 100.70%,99.31%, 101.10%,100.03% and 99.63%(all RSD <2%,n=9). Among 3 batches of Huaihua san samples ,the contents of above 5 components were 20.055-22.615,25.557-27.806,11.428-13.250,0.350-0.478,2.372-4.011 mg/g,respectively. CONCLUSIONS : Established method is simple ,accurate and reproducible ,and could be used for the simultaneous determination of 5 components in Huaihua san.
6.Effects of Different Processed Products of Whitmania pigra on Hemorheology and Coagulation Indexes in Acute Blood Stasis Model Rats
Changlin WANG ; Zhufeng CONG ; Guofei LIU ; Shaoping WANG ; Zedong XIANG ; Pingping DONG ; Ping SUN ; Hongjin ZHAO ; Peng GAO ; Long DAI
China Pharmacy 2020;31(16):1984-1988
OBJECTIVE:To stud y the effects of different processed products of Whitmania pigra on hemorheology and coagulation indexes in acute blood stasis model rats. METHODS :SD rats were randomly divided into blank group ,model group , aspirin group ,W. pigra hang-dried product low- ,medium- and high-dose groups ,W. pigra talcum powder-ironed product low- , medium- and high-dose groups ,W. pigra wine bran-processed product low- ,medium- and high-dose groups ,with 6 rats in each group. Except for blank group ,other groups received subcutaneous injection of epinephrine hydrochloride and ice water bath for 15 d to induce acute blood stasis model. From the 8th day of modeling ,rats in aspirin group were given aspirin 0.2 g/kg intragastrically. Rats in each dose group of W. pigra processed products were given relevant medicine 0.35,1.4,3.5 g/kg intragastrically(calculated by crude drug ). Rats in blank group and model group were given constant volume of normal saline intragastrically, once a day , for consecutive 8 days. Hemorheology indexes as whole blood viscosity (high, medium and low shearrate ),plasma viscosity ,erythrocyte com deformation index ,erythrocyte aggregation index ,hematocrit, and blood coagulation indexes as prothrombin time (PT), mail:wcl19960125@163.com activated partial prothrombin time (APTT),thrombin time (TT)were determined. RESU LTS:Compared with blank group ,whole blood viscosity under different shear rates ,plasma viscosity , erythrocyte aggregation index and hematocrit of model group were increased significantly ,while erythrocyte deformation index was significantly decreased ,PT,TT and APTT were significantly shortened (P<0.01). Compared with model group ,whole blood viscosity under different shear rates ,plasma viscosity ,erythrocyte aggregation index and hematocrit of aspirin group and W. pigra hang-dried product ,talcum powder-ironed product ,wine bran-processed product high-dose groups were decreased significantly , while erythrocyte deformation index were significantly increased ,and PT (only W. pigra talcum powder-ironed products high-dose group),APTT(except for W. pigra hang-dried products high-dose group )and TT were prolonged significantly. The whole blood viscosity of W. pigra hang-dried product medium-dose group under low shear rate ,and those of W. pigra talcum powder-ironed product low-dose ,wine bran-processed product medium-dose groups under low and medium shear rates were decreased significantly. Erythrocyte deformation index of W. pigra talcum powder-ironed product medium-dose group was increased significantly ,while erythrocyte aggregation index was decreased significantly ,and PT ,TT were prolonged significantly. APTT of W. pigra hang-dried product medium-dose group was prolonged significantly. Hematocrit of W. pigra wine bran-processed product low-dose group was decreased significantly (P<0.05 or P<0.01). CONCLUSIONS : W. pigra hang-dried, talcum powder-ironed and wine bran-processed product can effectively improve hemorheology indexes and prolong blood coagulation time.
7.Content Determination of 4 Indicator Components in Shengyu Decoction Lyophilized Powder by HPLC
Danhua DONG ; Yujun LIU ; Yanan LI ; Xianghao HU ; Ping SUN ; Ting LI ; Juyan LIU ; Peng GAO
China Pharmacy 2020;31(5):576-580
OBJECTIVE:To establish the content determin ation method of ferulic acid ,verbascoside,ligustilide and astragaloside in Shengyu decoction lyophilized powder. METHODS :HPLC method was adopted to determine 4 components in 3 batches of lyophilized powder. The determination of ferulic acid ,verbascoside and ligustilide was performed on Inertsil ODS-SP C 18 column with mobile phase consisted of methanol- 0.1% phosphoric acid (gradient elution )at the flow rate of 1.0 mL/min;detector was diode array detector ;detection wavelength was set at 330 nm;column temperature was 30 ℃,the sample size was 10 μL. The determination of astragaloside was performed on Kromasil C 18 column with mobile phase consisted of acetonitrile-water (32∶68,V/ V);detector was evaporative light scattering detector ;the drift tube temperature wa s 100 ℃,the carrier gas (air)flow rate was 2.5 L/min at the flow rate of 1.0 mL/min;column temperature was 30 ℃,the sample size was 10 μL. RESULTS:The linear ranges of ferulic acid ,verbascoside,ligustilide and astragaloside were 0.050 15-10.03 μg(r=0.999 8),0.067 80-13.56 μg(r= 0.999 9),0.057 30-11.46 μg(r=0.999 5),1.128-11.28 μg(r=0.999 3),respectively. The detection limits were 2.12×10-4,1.30× 10-3,8.02×10-4,1.09×10-3 μg,respectively. The limit of quantification were 7.43×10-4,3.87×10-3,2.34×10-3,3.36×10-3 μg, respectively. RSDs of precision ,stability(12 h)and reproducibility tests were all lower than 2%(n=6). Average recovery rates were 99.6%(RSD=0.83%,n=6),100.9%(RSD=1.07%,n=6),98.8%(RSD=0.84%,n=6)and 101.3%(RSD=0.99%, n=6),respectively. The contents of ferulic acid ,verbascoside,ligustilide and astragaloside in 3 batches of samples were 1.225-1.248, 0.413-0.424, 0.325-0.332, 0.394-0.404 mg/g, respectively (RSDs among batches were lower than 1.5% ). CONCLUSIONS:Established method is stable ,reproducible,rapid and accurate for the content determination of ferulic acid , verbascoside, ligustilide and astragaloside in Shengyu
8.Influencing factors of medical exposure frequency for community residents
Yuli CAO ; Yinping SU ; Hui XU ; Quanfu SUN ; Jinsheng CHENG
Chinese Journal of Radiological Health 2023;32(2):119-124
Objective To obtain the medical exposure frequency data of residents through a questionnaire survey on mobile phone networks in the community, and to analyze the main influencing factors. Methods A snowball sampling method was used to expand the sampling from medical staff in a county hospital to the community. The questionnaire included personal information such as occupation category, type of medical insurance, and education level as well as basic radiation-related knowledge and medical exposure type and frequency of respondents received in 2021. A logistic regression model was used to analyze the influencing factors of medical exposure. Results A total of 223 respondents were investigated, and 12.56% of them received radiological diagnosis and radiotherapy in 2021. The proportion of males receiving medical exposure was significantly higher than that of females (19.64% vs 10.18%). The likelihood of the person receiving medical exposure increased considerably with age, with the proportion of residents receiving medical exposure at 20, 30, 40, and 50 years of age being 6.25%, 7.95%, 19.23%, and 33.33%, respectively. Of persons who received medical exposure, 57.14% underwent CT scans. Persons participating in the new rural cooperative medical system were likely less to receive radiological diagnosis and radiotherapy than urban workers and residents (OR = 0.55, CI:0.13-2.23, P > 0.05). Conclusion Questionnaire survey in the community can obtain more information about the medical exposure of residents, and age and type of medical insurance are important factors affecting the frequency of medical exposure.
9.UHPLC-Q-Exactive Orbitrap MS/MS-based rapid identification of chemical components in substance benchmark of Kaixin San.
Hao-Ran LI ; Ping-Ping DONG ; Hua-Jian LI ; Jing XU ; Hong WANG ; Yi-Fang CUI ; Zhi-Qiang SUN ; Peng GAO ; Jia-Yu ZHANG
China Journal of Chinese Materia Medica 2022;47(4):938-950
Ultra-performance liquid chromatography-quadrupole-electrostatic field Orbitrap mass spectrometry(UHPLC-Q-Exactive Orbitrap MS/MS) was used for rapid identification of the chemical components in Kaixin San substance benchmark. The gradient elution was performed through a Waters ACQUITY~(TM) BEH C_(18) column(2.1 mm×150 mm, 1.7 μm) with water-acetonitrile as mobile phase, a column temperature of 30 ℃, a flow rate of 0.3 mL·min~(-1), and a sample size of 1 μL. The scanning was performed in the negative ion mode. The complex component groups in Kaixin San substance benchmark were quickly and accurately identified and clearly assigned based on the comparison of the retention time and MS data with those of the reference substance as well as the relative molecular weight of the same or similar components in the mass spectrum database and literature. A total of 77 compounds were identified, including 26 saponins, 13 triterpenoid acids, 20 oligosaccharide esters, 5 xanthones, and 13 other compounds. The qualitative method established in this study can systematically, accurately, and quickly identify the chemical components in Kaixin San substance benchmark, which can provide a basis for the further analysis of its active components in vivo and the establishment of its quality control system.
Benchmarking
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Chromatography, High Pressure Liquid/methods*
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Drugs, Chinese Herbal/chemistry*
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Tandem Mass Spectrometry/methods*
10.Analysis of gastric gastrointestinal stromal tumors in Shandong Province: a midterm report of multicenter GISSG1201 study.
Qingsheng HOU ; Wenqiang LUO ; Leping LI ; Yong DAI ; Lixin JIANG ; Ailiang WANG ; Xianqun CHU ; Yuming LI ; Daogui YANG ; Chunlei LU ; Linguo YAO ; Gang CUI ; Huizhong LIN ; Gang CHEN ; Qing CUI ; Huanhu ZHANG ; Zengjun LUN ; Lijian XIA ; Yingfeng SU ; Guoxin HAN ; Xizeng HUI ; Zhixin WEI ; Zuocheng SUN ; Hongliang GUO ; Yanbing ZHOU
Chinese Journal of Gastrointestinal Surgery 2017;20(9):1025-1030
OBJECTIVETo summarize the treatment status of gastric gastrointestinal stromal tumor (GIST) in Shandong province,by analyzing the clinicopathological features and prognostic factors.
METHODSClinicopathological and follow-up data of 1 165 patients with gastric GIST between January 2000 and December 2013 from 23 tertiary referral hospitals in Shandong Province were collected to establish a database. The risk stratification of all cases was performed according to the National Institutes of Health(NIH) criteria proposed in 2008. Kaplan-Meier method was used to calculate the survival rate. Log-rank test and Cox regression model were used for univariate and multivariate prognostic analyses.
RESULTSAmong 1 165 cases of gastric GIST, 557 were male and 608 were female. The median age of onset was 60 (range 15-89) years. Primary tumors were located in the gastric fundus and cardia in 623 cases(53.5%), gastric body in 346 cases(29.7%), gastric antrum in 196 cases(16.8%). All the cases underwent resection of tumors, including endoscopic resection (n=106), local resection (n=589), subtotal gastrectomy(n=399), and total gastrectomy(n=72). Based on the NIH risk stratification, there were 256 cases (22.0%) at very low risk, 435 (37.3%) at low risk, 251 cases (21.5%) at intermediate risk, and 223 cases (19.1%) at high risk. A total of 1 116 cases(95.8%) were followed up and the median follow-up period was 40 (range, 1-60) months. During the period, 337 patients relapsed and the median time to recurrence was 34 (range 1-60) months. The 1-, 3-, and 5-year survival rates were 98.6%, 86.1% and 73.4%, respectively. The 5-year survival rates of patients at very low, low, intermediate, and high risk were 93.1%, 85.8%, 63.0% and 42.3% respectively, with a statistically significant difference (P=0.000). Multivariate analysis showed that primary tumor site (RR=0.580, 95%CI:0.402-0.835), tumor size (RR=0.450, 95%CI:0.266-0.760), intraoperative tumor rupture(RR=0.557, 95%CI:0.336-0.924), risk classification (RR=0.309, 95%CI:0.164-0.580) and the use of imatinib after surgery (RR=1.993, 95%CI:1.350-2.922) were independent prognostic factors.
CONCLUSIONSThe choice of surgical procedure for gastric GIST patients should be based on tumor size. All the routine procedures including endoscopic resection, local excision, subtotal gastrectomy and total gastrectomy can obtain satisfactory curative outcomes. NIH classification has a high value for the prediction of prognosis. Primary tumor site, tumor size, intraoperative tumor rupture, risk stratification and postoperative use of imatinib are independent prognostic factors in gastric GIST patients.