1.Tolerance of rat bone marrow mesenchymal stem cells overexpressing human heme oxygenase 1 to ischemia/hypoxia injury
Ning-Bo DENG ; Teng-Long HAN ; Yuan-Qing ZENG ; Zhi-Xin JIANG
Chinese Journal of Tissue Engineering Research 2017;21(29):4617-4622
BACKGROUND:Under ischemia/hypoxia microenvironment,very low survival rate of transplanted bone marrow mesenchymal stem cells (BMSCs) in the host limits its efficacy in the treatment of acute myocardial infarction.OBJECTIVE:To explore the tolerance of human heme oxygenase 1 (hHO-1) gene modified rat BMSCs to ischemia/hypoxia injury.METHODS:The rat BMSCs were transfected with hHO-1 recombinant adenovirus.Western blot assay was used to determinate the optimal time of hHO-1 protein expression.hHO-1 modified rat BMSCs were cultured in hypoxia and serum-free conditions that simulated ischemia/hypoxia microenvironment in vivo.Cell counting kit-8 and trypan blue staining were performed to detect the survival rate of BMSCs at 12,24,48,72 hours after culture under the ischemia/hypoxia microenvironment.Flow cytometry was used to detect apoptosis in BMSCs at 24 hours after culture under the ischemia/hypoxia microenvironment.RESULTS AND CONCLUSION:The expression of hHO-1 protein was highest at 4 days after transfection.Under the ischemia/hypoxia microenvironment for 12,24,48,72 hours,the survival rates of transfected BMSCs were significantly higher as compared with the untransfected cells (P < 0.05),shown by the cell counting kit-8 and trypan blue staining.In addition,the results from flow cytometry showed that there was a higher survival rate of transfected BMSCs than untransfected cells at 24 hours of culture under the ischemia/hypoxia microenvironment (P < 0.05).To conclude,hHO-1 modified rat BMSCs have stronger tolerance to the ischemia/hvpoxia microenvironment.
2.Efficacy of minimally invasive left ventricular epicardial lead placement for cardiac resynchronization therapy.
Yun-long WANG ; Xue-jun REN ; Wen-bin LI ; Jin-sheng XIE ; Teng-yong JIANG ; Zhi-hong HAN ; Fang CHEN ; Ji-hong GUO
Chinese Journal of Cardiology 2010;38(7):614-617
OBJECTIVEVentricular resynchronization might be achieved via minimally invasive left ventricular epicardial lead placement.
METHODSix patients with congestive heart failure underwent minimally invasive left ventricular epicardial lead placement after failed coronary sinus cannulation were followed up for 1 year, cardiac function and LV lead threshold were evaluated.
RESULTSThere were no in-hospital deaths, intraoperative complications and diaphragm stimulation. Correct lead positioning was achieved in all 6 patients. LV lead thresholds remained unchanged [(1.2 ± 0.5) V vs (1.1 ± 0.4) V, P = 0.68] at 12 months follow-up. Improvements on 6 min walking test [(327 ± 77) m vs (267 ± 68) m, P = 0.001], LVEF [(26.1 ± 6.0)% vs (38.2 ± 4.7)%, P = 0.004], and NYHA functional class were evidenced at 12 months follow-up.
CONCLUSIONMinimally invasive left ventricular epicardial lead placement is a safe and reliable technique and should be considered as an alternative option in case of difficult coronary venous anatomy and inability to position the lead for resynchronization therapy.
Adult ; Cardiac Pacing, Artificial ; methods ; Cardiac Resynchronization Therapy ; Female ; Heart Failure ; surgery ; Heart Ventricles ; surgery ; Humans ; Male ; Middle Aged ; Pericardium ; surgery
3.Transfection efficiency of three different lentiviruses with green fluorescent protein gene to transfect rabbit bone marrow mesenchymal stem cells
Yu XIAO ; Yan-Lin LI ; Kun WANG ; Long-Teng LI ; Xiang-Jia YAN ; Xu-Han MENG
Chinese Journal of Tissue Engineering Research 2017;21(29):4642-4647
BACKGROUND:We explore the transfection effects of different lentiviruses on bone marrow mesenchymal stem cells (BMSCs) from the same source in order to screen out appropriate cell transfection tools.OBJECTIVE:To explore the difference in the transfection efficiency of three different lentiviruses carrying green fluorescent protein (GFP) to transfect the rabbit BMSCs.METHODS:Bone marrow samples were extracted from the ilium of 2-week-old Japan white rabbits,and then the BMSCs were separated,purified and extended using density gradient centrifugation.Cell surface markers were identified using flow cytometry.pTomo-GFP lentivirus,Ad-GFP lentivirus and lenti-GFP lentivirus were used to transfect the BMSCs,and the transfection efficiency was tested using flow cytometry.The cell viability was detected using cell counting kit-8 before and after transfection.RESULTS AND CONCLUSION:As the positive rates of CD44,CD34 and CD45 were 75.64%,6.14% and 9.48%,respectively,the cultured cells were identified as BMSCs by the flow cytometry.The expression levels of GFP after transfection by pTomo-GFP,Ad-GFP and ienti-GFP were 2.64%,16.72% and 45.24%,respectively.The cell growth curve showed that pTomo-GFP lentivirus,Ad-GFP and ienti-GFP lentivirus showed certain effects on cell growth,pTomo-GFP and Ad-GFP had an effect on cell growth at incubation period and logarithmic growth phase at 1-8 days after transfection,which may cause some interference to the viability of early cells,ienti-GFP had little effect on cell growth in the short term,but there was a change in cell growth at day 12 after lenti-GFP transfection,and a long-term effect of lenti-GFP on cell viability was considered.Therefore,lenti-GFP lentivirus could highly transfect the BMSCs and could not impact the normal growth of BMSCs,indicating that lenti-GFP lentivirus is suitable for BMSCs tracing and gene transfection.
4.Analysis on intestinal absorption of paeoniflorin lipid liquid crystalline nanoparticles via everted intestinal sacs.
Shuang TENG ; Ling QIU ; Hong-Ge CHEN ; Bao-de SHEN ; Cheng-Ying SHEN ; Ping-Hua XU ; Jin HAN ; Hai-Long YUAN
China Journal of Chinese Materia Medica 2016;41(19):3674-3678
To study the absorption kinetics of paeoniflorin lipid liquid crystalline nanoparticles (Pae-LLCN) in different intestinal segments of rats and compare them with paeoniflorin(Pae) solution. Rat everted gut sac models were adopted for intestinal absorption test, and Pae content was determined by HPLC method to study the absorption characteristics of Pae-LLCN in rat duodenum, jejunum, ileum and colon, and investigate the effects of different drug concentrations on intestinal absorption. Results showed that Pae-LLCN and Pae were well absorbed at different intestine segments and different concentrations. The absorption constant Ka was increased with the increasing of the drug concentration, indicating possible passive absorption. The accumulative absorption volume Q and absorption constant Ka of Pae-LLCN were higher than those of Pae at each intestinal segment(P<0.05). The results revealed that Pae-LLCN and Pae could be well absorbed in whole intestinal segments and its mechanism may be passive absorption. LLCN can effectively improve the intestinal absorption of Pae.
5.Application Prospect of Integrative Omics in Forensic Identification of Methamphetamine-Associated Psychosis.
Long-Rui ZHAO ; Jian-Bo ZHANG ; Wei HAN ; Li ZHU ; Teng CHEN ; Fang-Lin GUAN
Journal of Forensic Medicine 2022;38(5):650-656
The clinical symptoms and signs of methamphetamine-associated psychosis (MAP) and schizophrenia are highly similar, but the situation is completely different when MAP and schizophrenia patients need to be assessed for criminal responsibility after they comitted a harmful behavior. Therefore, the distinction between the two psychoses is very important in forensic psychiatry. At present, the identification of these two psychoses is mainly dependent on the corresponding criteria such as the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) and the Chinese Classification of Mental Disorders Version 3 (CCMD-3). It's challenging to diagnose and distinguish between the two in practical cases due to their similar clinical symptoms and the lack of effective objective indexes. Different from the limitations of single omics, integrative omics intergrates data from multiple dimensions and has been extensively studied in the field of schizophrenia and has achieved some preliminary results. In view of the correlation between MAP and schizophrenia and the potential application value of integrative omics, this paper proposes an integrative omics strategy for MAP pathogenesis and forensic identification, aiming to improve the further understanding of the relationship between the two psychoses and the corresponding pathogenesis. It also provides references for the future exploration of integrative omics in forensic precise identification and effective monitoring and early warning methods.
Humans
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Methamphetamine/adverse effects*
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Psychoses, Substance-Induced/etiology*
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Psychotic Disorders/genetics*
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Schizophrenia/genetics*
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Diagnosis, Differential
6.Analysis and Identification of Chemical Constituents of Citri Sarcodactylis Fructus by UPLC-Q-Orbitrap HRMS
Han-wen HU ; Yong-yan ZHAO ; Tian-long YANG ; Zhen-xing ZHENG ; Teng PENG ; Fang DENG
Chinese Journal of Experimental Traditional Medical Formulae 2020;26(7):148-155
Objective::To analyze and identify the chemical constituents of Citri Sarcodactylis Fructus by an ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap high-resolution mass spectrometry (UPLC-Q-Orbirap HRMS) method. Method::Compounds were separated on Thermo Scientific Accucore™ C18 column (3 mm×100 mm, 2.6 μm). The mobile phase was 0.1% formic acid solution and 0.1% formic acetonitrile solution. The flow rate was 0.3 mL·min-1, and the column temperature was set at 30 ℃. HRMS was performed using an electrospray ion source (ESI), and scanned in a positive ion mode by means of full scan/data dependent secondary scan (Full MS/dd-MS2). Compound Discoverer 3.0 software that could be linked to mzCloud network database and local high-resolution mass spectrometry database of traditional Chinese medicine components was used to analyze the data, based on accurate molecular mass, retention behaviors and characteristic ion fragmentation of the compounds, as well as literature information and relevant reference materials. Result::Totally 54 chemical constituents in Citri Sarcodactylis Fructus were identified, including 16 flavonoids, 17 coumarins, 3 limonoids, 6 nucleosides and nucleobases, 2 organic acids, 3 aromatic aldehydes, 1 amino acid and 6 other components. Conclusion::The established UPLC-Q-Orbitrap HRMS method can be used to effectively and rapidly identify main chemical constituents of Citri Sarcodactylis Fructus. The findings provide a methodological reference and theoretical foundation for defining the pharmacodynamic material base and optimizing quality control index of Citri Sarcodactylis Fructus, which is of guiding significance for further development and utilization of the resources.