1.Establishment,identification and application of induction culture method of mono-nuclear macrophages from cow bone marrow
Yu WANG ; Xiaolin YANG ; Lili GUO ; Pengfei GONG ; Jingze WU ; Wei MAO ; Shuangyi ZHANG ; Bo LIU ; Jinshan CAO
Chinese Journal of Veterinary Science 2024;44(8):1674-1681
In order to establish the isolation,culture and identification method of cow bone marrow-derived macrophages,three different media(RPMI-1640,DMEM,DMEM/F12)were added with 20%fetal bovine serum(FBS),2.4%chlorine-streptomycin,1.2%glutamine(Gln),and M-CSF(20 ng/mL),respectively,to induce the monocytes extracted from the bone marrow of dairy cows to become macrophages.The induced M0 macrophages were polarized into M1-type macrophages by adding lipopolysaccharide(LPS).The morphology of macrophages was observed by optical mi-croscope at day 1,4 and 7,and the differences of differentiated macrophages between the three media were compared.The effects of prostaglandin D2(PGD2)-DP2 receptor pathway on the secre-tion of cytokines(IL-6,TNF-α)induced by Escherichia coli and phagocytosis of macrophages were also investigated.The results showed that the morphological changes of cells cultured in the medium of RPMI-1640 were the most obvious and the number was large.A large number of char-acteristic markers of mononuclear macrophages were detected(M0 markers:CD1 1b,CD14;M1 markers:CD11b,CD80)expression,M0 and M1 macrophage purity were 79.9%and 93.5%,re-spectively.COX-2 and H-PGDS gene expressions were significantly increased in E.coli group com-pared with the blank control group.The secretion of PGD2also increased significantly(P<0.000 1).DP2 receptor inhibitors(CAY10471,CAY10595)could significantly inhibit the secretion of E.coli in-duced pro-inflammatory cytokines(IL-6,TNF-α)and significantly enhance the killing effect of macrophages on E.coli.The above results showed that the induced cells had the characteristic mor-phology and immunophenotype of macrophages.E.coli can induce the production of PGD2 in mac-rophages,and the PGD2-DP2 pathway regulates the secretion of cytokines in E.coli infected macro-phages.
2.The role of first-aid network construction in the early treatment of patients with critically severe hydrofluoric acid burns
Yuanhai ZHANG ; Pengfei TIAN ; Wei ZHANG ; Chunjiang YE ; Shulei MAO ; Chunmao HAN ; Jianfen ZHANG ; Xingang WANG
Chinese Journal of Burns 2021;37(10):921-928
Objective:To explore the role of first-aid network construction in the early treatment of patients with critically severe hydrofluoric acid burns.Methods:Twenty-seven fluorine chemical enterprises distributed in Zhejiang province, Jiangxi Province, Fujian Province, and Inner Mongolia Autonomous Region and 22 hospitals with burn/plastic department or professional burn treatment group in Zhejiang province, including Zhejiang Quhua Hospital, and 5 hospitals outside Zhejiang province were involved in the first-aid network construction as member units. As the main unit, Zhejiang Quhua Hospital was responsible for the daily maintenance and technical guidance of the first-aid network. Zhejiang Quhua Hospital was assigned as the designated emergency hospital for 20 fluorine chemical enterprises, a near emergency hospital to the other 7 fluorine chemical enterprises was assigned as the designated hospital for them. Medical records of 56 patients (all males) with critically severe hydrofluoric acid burns who admitted to 5 first-aid network hospitals from January 2006 to June 2021, meeting the inclusion criteria, were involved in the retrospective cohort study. Based on whether the enterprise belonging to the first-aid network construction or not, the patients were divided into first-aid network group (27 cases, aged (41±9) years) and non first-aid network group (29 cases, aged (42±10) years). After the patients in the first-aid network group were injured, the enterprises and hospitals linked up immediately. The hospital where the patient was treated mobilize the treatment force, equipment, materials, and drugs in advance by the first-aid network, thereby realizing seamless joint between pre-hospital first-aid and in-hospital treatment. The hospital started the first-aid process and temporarily mobilized the rescue forces, equipment, materials, and drug after patients in non first-aid network group arrived at the department of emergency of the hospital. The time from injury to medical service, the first detection time of serum calcium, the time staying in department of emergency, the duration of hypocalcemia and hypomagnesemia, and the treatment outcome of patients in the two groups were recorded. Data were statistically analyzed with chi-square test, Fisher's exact probability test, independent-sample t test, and Wilcoxon rank-sum test. Results:The time from injury to medical service, the first detection time of serum calcium, and the time staying in department of emergency of patients in first-aid network group were 40.0 (30.0, 55.0), 23.0 (17.5, 37.5), and 42.0 (37.0, 53.0) min, which were significantly shorter than 180.0 (120.0, 240.0), 31.0 (22.5, 47.5), 61.0 (52.0, 65.5) min in non first-aid network group ( Z=-6.17, -1.98, -4.15, P<0.05 or P<0.01). The duration of hypocalcemia and hypo- magnesemia of patients in first-aid network group were 1.2 (1.1, 1.6) and 1.9 (1.7, 2.1) h, which were significantly shorter than 4.6 (3.1, 6.2) and 3.2 (2.5, 4.6) h in non first-aid network group ( Z=-5.80, -4.81, P<0.01). Three patients (11.1%) in first-aid network group died, among whom 2 patients died at 40 min after injury and 1 patient died 9.0 h after injury. Four patients (13.8%) died in non first-aid network group at 3.0, 3.0, 4.5, and 7.0 h after injury, respectively. The mortality rates of patients in the two groups were similar ( P>0.05). Conclusions:Critically severe hydrofluoric acid burn is an extremely urgent situation encountered in clinical practice. The construction of a first-aid network creates condition for on-site treatment of patients and improves the first-aid efficiency, thereby gaining time to save lives.
3.Effects of radiation on echinococcus in vitro culture and its mRNA expression of Gadd45α
Pengfei LU ; Ge WU ; Yi XIONG ; Tingting DANG ; Yongxing BAO ; Rui MAO
Chinese Journal of Endemiology 2020;39(6):391-395
Objective:To investigate the killing effect of radiation on echinococcus in vitro culture and its effect on the mRNA expression of growth arrest and DNA damage 45 alpha (Gadd45α) gene. Methods:Echinococcus from naturally infected sheep liver was cultured in vitro and divided into 7 groups. The echinococcus was irradiated with 6 MeV at doses of 0 (control), 20, 40, 60, 80, 100, and 120 Gy, respectively. The growth of echinococcus was observed under light microscope at 1, 3, 5 and 7 d after the radiation. The expression of Gadd45α mRNA in control, 20, 40 and 60 Gy groups of echinococcus was detected by RT-PCR technique at 7 d after the radiation. Results:The disintegration and exfoliation of echinococcus were observed under light microscope at 1, 3, 5 and 7 d after the radiation, and the death rate of echinococcus was positively correlated with the radiation dosages ( r = 0.81, P < 0.05). After the radiation at 7 d, compared with the control group (100.00 ± 0.00), the mRNA expression levels of Gadd45α in echinococcus of 20, 40, and 60 Gy groups were significantly increased (279.74 ± 80.08, 759.38 ± 160.98, 1 666.68 ± 316.36, P < 0.01), and it was positively correlated with the radiation dosages ( r = 0.93, P < 0.01). Conclusion:Radiation has a certain killing effect on echinococcus cultured in vitro, and there is a certain dose-effect relationship with the radiation dosages, and Gadd45α gene may be involved in the molecular mechanism of radiation-induced killing of echinococcus in vitro.
4. Follow-up of people living with HIV/AIDS by primary health care institutions in rural area of Jiangxi province
Pengfei FAN ; Qing YANG ; Yurong MAO ; Qiang HU ; Houlin TANG ; Jian LI ; Yaling LUO ; Fen WANG ; Huanqing ZHAN ; Siming ZANG
Chinese Journal of Epidemiology 2019;40(3):346-349
Objective:
To understand the current status of follow up of people living with HIV/AIDS by health service at grass root in rural area of Jiangxi province and related factors, and provide references for the promotion of the follow up by grass root health service.
Methods:
People living HIV/AIDS aged ≥18 years and diagnosed before 31 December 2017 in 6 townships of Xinjian, Yushan counties and Guixi city were included in the study in Jiangxi province. They had been followed up for more than one time after the first epidemiologic survey. The information about their demographic characteristics and HIV infection status were collected by using self-designed questionnaire. Univariate and multivariate logistic regression analyses were conducted to identify the factors that influencing the acceptance of follow up by grass root health service.
Results:
Of the 373 surveyed HIV infected subjects aged (53.06±16.15) years, 261 were males (70.0
5. The situation and related factors of satisfaction of HIV-infected people towards the follow-up management of primary medical and health care institutions in rural areas of Jiangxi Province
Pengfei FAN ; Qing YANG ; Yurong MAO ; Houlin TANG ; Jian LI ; Fen WANG ; Huanqing ZHAN ; Siming ZANG
Chinese Journal of Preventive Medicine 2019;53(5):522-526
The HIV-infected people were investigated for their satisfactory situation towards the follow-up management of primary medical and health care institutions in Xinjian County, Yushan County and Guixi City of Jiangxi Province from January to July 2018 and related factors were also analyzed. The total score of the Infected Patients′ Follow-up Management Satisfaction Scale higher than 59 was defined as the satisfactory. 75.68% of 259 infected patients were satisfied with the follow-up management. Compared to farmers and other follow-up modes, non-farmers (
6.Retraction: Differential expression of exosomal miRNAs in osteoblasts in osteoarthritis Journal of Central South University. Medical Science, 2018, 43(12):1294-1300. DOI: 10.11817/j.issn.1672-7347.2018.12.003.
Bohao LIU ; Pengfei WU ; Lin MEI ; Yong LUO ; Hongxing LI ; Xinzhan MAO
Journal of Central South University(Medical Sciences) 2019;44(2):149-149
The article entitled "Differential expression of exosomal miRNAs in osteoblasts in osteoarthritis" published on Journal of Central South University (Medical Science), in Volume 43, Issue 12, 2018 (DOI: 10.11817/j.issn.1672-7347.2018.12.003) may have an unclear risk of bias due to insufficient understanding for some results. Further experimental studies are needed. We all agree to retract this article, and apologize to the Journal and readers for the possible negative impact.
7.The situation and related factors of satisfaction of HIV?infected people towards the follow?up management of primary medical and health care institutions in rural areas of Jiangxi Province
Pengfei FAN ; Qing YANG ; Yurong MAO ; Houlin TANG ; Jian LI ; Fen WANG ; Huanqing ZHAN ; Siming ZANG
Chinese Journal of Preventive Medicine 2019;53(5):522-526
The HIV?infected people were investigated for their satisfactory situation towards the follow?up management of primary medical and health care institutions in Xinjian County, Yushan County and Guixi City of Jiangxi Province from January to July 2018 and related factors were also analyzed. The total score of the Infected Patients′Follow?up Management Satisfaction Scale higher than 59 was defined as the satisfactory. 75.68% of 259 infected patients were satisfied with the follow?up management. Compared to farmers and other follow?up modes, non?farmers (OR=10.72, 95%CI: 2.07-55.63), and patients receiving follow?up service in responsible institutions ( OR=6.44, 95%CI : 3.12-13.30) were more satisfied with follow?up management.
8.The situation and related factors of satisfaction of HIV?infected people towards the follow?up management of primary medical and health care institutions in rural areas of Jiangxi Province
Pengfei FAN ; Qing YANG ; Yurong MAO ; Houlin TANG ; Jian LI ; Fen WANG ; Huanqing ZHAN ; Siming ZANG
Chinese Journal of Preventive Medicine 2019;53(5):522-526
The HIV?infected people were investigated for their satisfactory situation towards the follow?up management of primary medical and health care institutions in Xinjian County, Yushan County and Guixi City of Jiangxi Province from January to July 2018 and related factors were also analyzed. The total score of the Infected Patients′Follow?up Management Satisfaction Scale higher than 59 was defined as the satisfactory. 75.68% of 259 infected patients were satisfied with the follow?up management. Compared to farmers and other follow?up modes, non?farmers (OR=10.72, 95%CI: 2.07-55.63), and patients receiving follow?up service in responsible institutions ( OR=6.44, 95%CI : 3.12-13.30) were more satisfied with follow?up management.
9.Differential expression of exosomal miRNAs in osteoblasts in osteoarthritis.
Bohao LIU ; Pengfei WU ; Lin MEI ; Yong LUO ; Hongxing LI ; Xinzhan MAO
Journal of Central South University(Medical Sciences) 2018;43(12):1294-1300
To analyze the differentially expressed exosomal miRNAs in subchondral osteoblasts in patients with osteoarthritis (OA) and to investigate the key miRNAs potentially involved in the occurrence and progression of OA.
Methods: Subchondral bones were harvested from 6 patients with OA. All subjects were divided into two groups which was based on the severity of joint wear: An OA group, severely worn side of subchondral bone, and a control group, less worn side of subchondral bone. The exosomes were extracted from osteoblast cells and their characteristics were identified. Then exosomal miRNAs were extracted and sequencing analysis was conducted to compare the expression in the two groups. The most differentially expressed ones (log2Ratio≥2) were subject to miRNA target prediction and quantitative reverse transcription PCR (RT-qPCR) to further quantify the difference.
Results: Osteoblast extractions were confirmed to be exosomes, which were small double-membranous vesicles with 30-200 nm in diameter and 50-150 nm in peak value of particle size under the scanning microscope. High-throughput sequencing revealed 124 miRNAs whose expression significantly increased in the OA group. The most differentially expressed one with maximum fold change was hsa-miR-4717-5p and its target gene was RGS2. RT-qPCR demonstrated hsa-miR-4717-5p expression in the OA group was relatively higher than that in the control group (2.243 vs 0.480, P<0.01).
Conclusion: There is distinct difference in expression profiles of exosomal miRNAs in subchondral osteoblasts between patients with OA and normal subjects. Up-regulated expression of miRANs might participate in OA occurrance and progression.
Bone and Bones
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Exosomes
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genetics
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pathology
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Gene Expression Profiling
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Gene Expression Regulation
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Humans
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MicroRNAs
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genetics
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Osteoarthritis
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physiopathology
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Osteoblasts
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pathology
10.Targeted nano-vector for gene delivery into human bone marrow mesenchymal stem cells and cellular MR imaging in vitro
Pengfei PANG ; Bing LI ; Junjie MAO ; Bin ZHOU ; Xiaojun HU ; Yongyu ZHANG ; Feng AO ; Hong SHAN
Chinese Journal of Medical Imaging Technology 2017;33(10):1463-1469
Objective To explore the feasibility and efficacy of an MRI-visible,targeted,nano-vector which is synthesized by attaching a targeting ligand,the GD2 single chain antibody (scAb GD2),to the distal ends of PEG-g-PEI-SPION as a carrier for gene delivery into human bone marrow mesenchymal stem cells (hBMSCs) and in vitro cellular MR imaging.Methods scAbGD2-PEG-g-PEI-SPION was synthesized as previously reported.Gel electrophoresis was performed to assess the pDNA condensation ability of scAbGD2-PEG-g-PEI-SPION.The particle size and Zeta potential of scAbGD2-PEG-g-PEI-SPION/pDNA nanocomplexes were observed by dynamic light scattering.Cytotoxicity of scAbGD2-PEG-g-PEI-SPI-ON was evaluated by CCK-8 assay using hBMSCs.Gene transfection efficiency of scAbGD2-PEG-g-PEI-SPION in hBMSCs was quantified by flow cytometry,PEG-g-PEI-SPION,scAbGD2-PEG-g-PEI-SPION,scAbGD2-PEG-g-PEI-SPION+ free AbGD2 and scAbIgG2a-PEG-g-PEI-SPION group was established.The cellular internalization of scAbGD2-PEG-g-PEI-SPION/pDNA nanocomplexes was observed by confocal laser scanning microscopy and Prussian blue staining.MRI of scAbGD2-PEG-g-PEI-SPION was performed by cellular MRI scanning in vitro.Results scAbGD2-PEG-g-PEI-SPION condensed pDNA to form stable nanocomplexes of 80-100 nm in diameter and showed low cytotoxicity to hBMSCs.At the same N/P ratio,the transfection efficiency of scAbGD2-PEG-g-PEI-SPION group was significantly higher than those of other groups (P<0.001).At the optimal N/P ratio of 20,scAbGD2-PEG-g-PEI-SPION/pDNA obtained the highest transfection efficiency of (59.60 ± 4.50) % in hBMSCs.Furthermore,hBMSCs labeled with scAbGD2-PEG-g-PEI-SPION showed sensitive low signal intensity on MRI T2/T2 *-weighted images in vitro.Conclusion scAbGD2-PEG-g-PEI-SPION is an efficient MRL visible targeted nano vector for gene delivery into hBMSCs.

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