1.8-methoxsalen photochemistry enhances tumor cell immunogenicity by inducing ferroptosis in B16 cells
Yan ZHONG ; Yuwei LIN ; Wei CHEN ; Li TIAN ; Ling LI ; Zhong LIU
Chinese Journal of Blood Transfusion 2025;38(8):999-1007
Objective: To study the efficacy of photosensitizer 8- methoxsalen (8-MOP) combined with ultraviolet radiation A (UVA) in inducing ferroptosis in mouse melanoma B16 cells, and to assess the resultant changes in immunogenicity, and their impact on subsequent immune activation after treatment. Methods: 1) Mouse melanoma B16 cells were cultured and treated with 8-MOP (100 ng/mL) and UVA (4 J/cm
), and then cultured in a constant temperature incubator (37℃, 5%CO
) for 24 hours after irradiation. 2) CCK8 (cell proliferation and toxicity) detection kit was used to detect the death rate of tumor cells. 3) LPO (lipid peroxide) and GSH (glutathione) detection kits were used to detect the degree of oxidative damage of tumor cells; Changes of Fe
, mitochondrial membrane potential (JC-1) and BODIPY 581/591 C11 (lipid peroxidation detection kit) in tumor cells were detected by confocal microscope. Western blotting (WB) was performed to detect GPX4, SLC7A11 and NCOA4 to confirm ferroptosis. 4) The expression of HMGB1 (high mobility group protein 1), ATP and CRT (calreticulin) in the supernatant of tumor cell culture was detected by ELISA kit to evaluate the immunogenicity of tumor cells. 5) 1×10
B16 cells were injected subcutaneously into the skin of the back and neck of mice at a dose of 100 μL to construct a mouse melanoma model. Spleen mononuclear cells of tumor-bearing mice were extracted and immediately co-cultured with irradiated tumor cells for 48 h. Changes of dendritic cell (DC) maturity were detected by MHC-II, CD11c, CD80 and CD83 flow cytometry. Results: After UVAP, the survival rate of B16 cells decreased significantly (61.39±6.823 vs 84.81±7.026 vs 100.0±3.996, P<0.000 1, P<0.01). UVAP effectively induced ferroptosis in B16 cells, characterized by increased LPO and C11-bodipy lipid peroxidation, GSH depletion, Fe
accumulation, mitochondrial membrane depolarization, decreased GPX4 and SLC7A11 protein expression, and increased NCOA4 expression, all in line with the trend of ferroptosis. UVAP also enhanced tumor cell immunogenicity, evidenced by elevated release of ATP, CRT, and HMGB1. The immunogenicity of B16 cells increased, the expressions of ATP, CRT and HMGB1 increased, and the DC maturity increased (CD80: 31.92±4.071 vs 19.77±3.177; CD83: 21.40±4.787 vs 12.19±1.487, P<0.001, P<0.01). Conclusion: The combined action of 8-MOP and UVA can induce ferroptosis in B16 tumor cells, enhance the immunogenicity of tumor cells, release more tumor antigens, promote the maturation of DC, present antigens better, thereby facilitating subsequent immune activation.
2.An investigation on avian influenza virus distribution in poultry-related environment in Nanping city
Yating ZHANG ; Jingjing WU ; Qi LIN ; Yuwei WENG
Chinese Journal of Experimental and Clinical Virology 2024;38(2):138-143
Objective:To profile the distribution of avian influenza virus in poultry-related environment in poultry industry developed area in Fujian province, an investigation was conducted in Nanping city from Dec.2021 to Dec.2023.Methods:The samples from multiple types of external environment related to poultry in Nanping city were collected from Dec. 2021 to Dec. 2023, and the real-time fluorescence quantitative RT-PCR used to detect and subtype the influenza A virus (FluA). SPSS 26.0 software was used to analyze the distribution characteristics of FluA in poultry-related environment and the differences in time, places and sample types.Results:The overall positive rate of FluA in samples from poultry-related environment was 49.16% (1 435/2 919). The positive rates of H3, H5, H9 and H10 subtypes were 0.72% (21/2 919), 9.42% (275/2 919), 33.20% (969/2 919), 0.89% (26/2 919) respectively, and no H7 subtype was detected. The positive rate of mixed type (more than one subtype of FluA detected in a same sample) was 6.51% (190/2 919), and the positive rate of unknown subtype (positive for FluA but negative for H3/5/7/9/10) was 11.58% (338/2 919). The higher positive rate of FluA mainly occurred in autumn-winter season (September to February of the following year). In live poultry markets and slaughterhouses, the positive rates of FluA, H9 subtype, mixed type and unknown subtype were significantly higher than that in poultry farms. The positive rate of FluA in poultry drinking water and feces was higher than samples of other types, most of the positive samples were H9 subtype.Conclusions:The positive rate of FluA in poultry-related environment in Nanping city was higher in autumn-winter season. The investigation also showed that higher FluA positive rate in drinking water and feces sample and diversity of the virus existed in the place of multiple types of poultry clustered, such as live poultry markets and slaughterhouses.
3.Correlation analysis of robotic autonomous vertebral puncture pressure and path accuracy based on animal specimens
Tong XING ; Junsheng HUANG ; Ming LI ; Yuxi LI ; Yuwei LIANG ; Zhen CHE ; Enming CHEN ; Lin HUANG
Chinese Journal of Orthopaedics 2024;44(15):1018-1024
Objective:To investigate the impact of peak pressure upon different bone densities during autonomous puncture by orthopedic robot on puncture path deviation and bone cement leakage.Methods:A spinal surgery robot system was designed for autonomous vertebral puncture and bone cement injection, and six porcine spine specimens were used for single-segment or double-segment simulated percutaneous vertebral augmentation surgery. The accuracy of puncture path (Gertzbein-Robbins grading), bone cement leakage classification, and peak bone drill pressure were measured to assess the accuracy of autonomous vertebral puncture and bone cement leakage in vertebral cortical and cancellous bone of different densities.Results:A total of 64 porcine vertebrae were simulated for puncture, among which 53 vertebrae were classified as Grade A, 8 as Grade B, and 3 as Grade C according to the Gertzbein-Robbins grading. The cortical bone pressure of Grade A vertebrae was 6.663±0.319 N which was lower than that of Grade B (8.348±0.418 N) and Grade C (11.500±0.600 N), with significant differences ( F=341.000, P<0.001). The cancellous bone pressure of Grade A, B, and C vertebrae were 3.660±0.317, 3.594±0.608, and 4.117±0.257 N, respectively, with no significant difference ( F=2.496, P=0.091). There were 40 cases of no leakage, 20 cases of Type I leakage (leakage into the surrounding vertebrae), and 3 cases of Type II leakage (leakage into the vertebral canal), with an overall leakage rate of 36% (23/64). The peak cortical bone pressure for no leakage, Type I, and Type II leakage was 6.638±0.301, 6.792±0.404, and 6.753±0.473 N, respectively, and the peak cancellous bone pressure was 3.634±0.279, 3.783±0.423, and 3.920±0.255 N, respectively, with no significant difference ( F=1.521, P=0.227; F=2.106, P=0.131). Conclusion:During the autonomous puncture process of the novel orthopedic robot, the accuracy of autonomous puncture path decreased when the puncture pressure through the cortical bone was high, and the probability of invading the pedicle increased. The puncture pressure of cortical and cancellous bone had no significant effect on the occurrence rate of bone cement leakage.
4.Meta-synthesis of qualitative research on the experience of self-management behavior changes for cancer patients
Wenwen LIN ; Mei XU ; Yun XIA ; Ning JIANG ; Yuwei MO ; Li WANG ; Li NING
Chinese Journal of Nursing 2024;59(7):865-873
Objective To systematically evaluate the self-management behavior transformation experience of cancer patients,providing a basis for the subsequent development of targeted self-management intervention measures.Methods Qualitative studies related to self-management behavior transformation experience of cancer patients from Cochrane Library,CINAHL,Embase,PubMed,Web of science,PsycINFO,and China National Knowledge Infras-tructure,VIP Database,Wanfang Database,and China Biomedical Literature Database were searched by computer,with a search deadline of July 2023.The literature quality evaluation was conducted using the qualitative research quality evaluation standards of the Evidence Based Health Care Center at the Joanna Briggs Institute in Australia,and the results were integrated using a pooled integration method.Results A total of 12 articles were included,and 50 research results were extracted and summarized into 14 new categories.4 integrated results were synthesized,including the forms of self-management behavior transformation,the characteristics of self-management behavior transformation,the driving factors of self-management behavior transformation,and the obstacles to self-management behavior transformation.Conclusion Medical staff should pay attention to the real experience of cancer patients'self-management behavior transformation.Targeted ability cultivation and psychological intervention can be carried out based on the characteristics and influencing factors of cancer patients'self-management behavior transformation,promoting their smooth transformation of self-management behavior.
5.Effects of application of innovative key performance indicator lean teaching management system in clinical medical laboratory
Yuwei DI ; Huaxin MAI ; Zhengkang LI ; Jinwei HUANG ; Chenglong LIN ; Ying LUO ; Yujing YANG ; Kaixuan YUAN ; Ge HUANG ; Wei HUANG ; Bing GU
Chinese Journal of Laboratory Medicine 2024;47(6):701-705
Objective:To evaluate the performance of key performance indicator (KPI) lean teaching management system in clinical medical laboratory.Methods:Combining lean teaching management in universities with KPI system of enterprises, an innovative KPI lean teaching management system was developed and applied in Clinical Medical Laboratory of Guangdong Provincial People′s Hospital (GDPH). A total of 106 teachers, who had worked from January 2021 to December 2022 in GDPH, joined this study. Teachers were divided into 8 professional groups. Firstly, we quantified the teaching workload by class hours, evaluated the teaching outcomes base on national, provincial, school, and college levels to assign different teaching points, and linked the teaching KPI scores to the evaluation, salary, and professional title of teachers. Then, we analyzed the overall teaching points and teaching points for each professional group (2021-2022). Finally, we asked teaching managers, teachers, and colleagues to evaluate the KPI lean teaching management system and compared the effects before and after the implementation of this system.Results:Compared with 2021, the teaching scores of 106 teachers increased significantly from 1.0 (0.2, 2.7) to 3.8 (2.3, 6.0) in 2022 ( Z=8.1, P<0.01). The teaching scores of clinical molecules, clinical coagulation, clinical immunology, clinical microbiology, Huifu laboratory, and clinical biochemistry group were significantly higher in 2022 than the scores they got in 2021 (all P<0.05). Compared with 2021, there were 3 new set up of educational reform projects, 2 submitted teaching articles, 3 new competition awards, 7 outstanding teachers, and 5 outstanding students in 2022. After application of KPI lean teaching management, the evaluation scores of teaching work by teaching managers, teachers, and colleagues are all significantly improved ( P<0.05). Conclusion:KPI lean teaching management system could effectively enhance teachers′ work initiative, improve teaching efficiency and outcome, and promote the teaching quality. Therefore, based on the performance of KPI lean teaching management system in our study, it is possible to realize its potential in terms of lean management in clinical medical laboratory.
6.Effects of lncRNA MIR22HG on proliferation,apoptosis and inflammatory response of rheumatoid arthritis fibroblast -like synoviocytes by sponge adsorption of miR - 22-5p
Zhou Yang ; Shudian Lin ; Yuwei Zhan ; Lu Xiao ; keying Fu ; Xiaodie Huang
Acta Universitatis Medicinalis Anhui 2023;58(3):405-412
Objective:
To explore the effects of long non-coding RNA (LncRNA) MIR22HG on proliferation,apoptosis and inflammatory response of rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLSs) and its molecular mechanism.
Methods:
Synovial tissue samples were collected from 37 RA patients and 30 joint trauma patients in our hospital,and the expression levels of MIR22HG and miR-22-5p in synovial tissue were detected by qRT-PCR. RA-FLSs in human was isolated ,cultured and identified in vitro. MIR22HG siRNA interference plasmid (si-MIR22HG) and its negative control plasmid (si-NC) ,miR-22-5p inhibitor and its negative control (inhibitorNC) were transfected into RA-FLSs respectively or simultaneously.The expression levels of MIR22HG and miR- 22-5p were detected by qRT-PCR. CCK-8 was used to detect the proliferation activity of cells in various groups. Annexin Ⅴ- FITC / PI was used to detect the apoptosis rates of cells in various groups.ELISA was used to detect the levels of TNF-α , IL-1 β and IL-6 in the supernatant of cells in various groups.Western blot was used to detect the protein expression levels of Bcl-2,Bax and Cleaved caspase-3 of cells in various groups.The targeting relationship between MIR22HG and miR-22-5p was verified by dual luciferase reporter gene assay.
Results :
Compared with joint trauma patients,the expression level of MIR22HG in synovial tissues of RA patients increased (P<0. 05) , while the expression level of miR-22-5p decreased (P<0. 05) .Interference with MIR22HG inhibited the proliferation activity of RA-FLSs,decreased the levels of TNF-α , IL-1 β and IL-6 in cell supernatant and the protein expression level of Bcl-2 in cells (P<0. 05) ,and increased the apoptosis rate,the expression level of miR-22-5p and the protein expression levels of Bax and Cleaved casepase-3 (P <0. 05 ) .However,inhibition of miR-22-5p expression reversed the effects of MIR22HG gene silencing on proliferation,apoptosis and inflammation of RA-FLSs (P<0. 05) .Dual luciferase reporting assay showed that miR-22-5p was a potential downstream miRNA target of MIR22HG.
Conclusion
MIR22HG is highly expressed in synovial tissues of RA patients,and it may promote the proliferation and the inflammatory response of RA-FLSs and inhibit cell apoptosis by down regulating the expression of miR-22-5p.
7.Lateral septum adenosine A2A receptors control stress-induced depressive-like behaviors via signal-ing to hypothalamus and habenula
Muran WANG ; Peijun LI ; Zewen LI ; SILVA S.da BEATRIZ ; Wu ZHENG ; Zhenghua XIANG ; Yan HE ; Tao XU ; CORDEIRO CRISTINA ; Lu DENG ; Yuwei DAI ; Mengqian YE ; Zhiqing LIN ; Jianhong ZHOU ; Xuzhao ZHOU ; Fenfen YE ; CUNHA A RODRIGO ; Jiangfan CHEN ; Wei GUO
Chinese Journal of Pharmacology and Toxicology 2023;37(7):547-548
Depressive disorder ranks as a major bur-den of disease worldwide,yet the current antidepressant medications are limited by frequent non-responsiveness and significant side effects.The lateral septum(LS)is thought to control of depression,however,the cellular and circuit substrates are largely unknown.Here,we identified a subpopulation of LS GABAergic adenosine A2A receptors(A2AR)-positive neurons mediating depres-sive symptoms via direct projects to the lateral habenula(LHb)and the dorsomedial hypothalamus(DMH).Activa-tion of A2AR in the LS augmented the spiking frequency of A2AR-positive neurons leading to a decreased activation of surrounding neurons and the bi-directional manipula-tion of LS-A2AR activity demonstrated that LS-A2ARs are necessary and sufficient to trigger depressive pheno-types.Thus,the optogenetic modulation(stimulation or inhibition)of LS-A2AR-positive neuronal activity or LS-A2AR-positive neurons projection terminals to the LHb or DMH,phenocopied depressive behaviors.Moreover,A2AR are upregulated in the LS in two male mouse mod-els of repeated stress-induced depression.This identifica-tion that aberrantly increased A2AR signaling in the LS is a critical upstream regulator of repeated stress-induced depressive-like behaviors provides a neurophysiological and circuit-based justification of the antidepressant poten-tial of A2AR antagonists,prompting their clinical transla-tion.
8.Distribution and transfusion efficacy of unexpected antibody
Rong ZHANG ; Kai KANG ; Jiang XIE ; Yuwei LIN ; Zujun LI ; Jiping XU ; Zhengqiu LIAN ; Ling LI
Chinese Journal of Blood Transfusion 2023;36(12):1132-1135
【Objective】 To investigate the clinical characteristics and antibody distribution as well as evaluate the transfusion efficacy in unexpected antibody positive patients. 【Methods】 A total of 12 235 patients from January 1, 2022 to March 31, 2023 who hospitalized in our hospital and applied for blood transfusion were selected, and those with unexpected antibody were included. The clinical data, including gender, age, diagnosis, blood type, history of transfusion and pregnancy were collected for antibody distribution analysis. Patients who received transfusion were grouped according to the DAT results and the components of red blood cells transfused, and the Hb values of each group before and after transfusion were compared. 【Results】 Among12 235 patients, 118 were positive for antibody screening, with a prevalence of 0.96%. The antibodies from Rh system were the most common (27.43%, 48/175), followed by MNS system (8.57%, 15/175) and Lewis system (6.29%, 11/175), mainly anti-E (18.29%, 32/175), anti-M (8.00%, 14/175) and anti-Lea (5.71%, 10/175). In addition, 62 transfused patients were divided into group A with suspended red blood cell transfusion and group B with washed red blood cell transfusion for positive DAT, and group C for negative DAT. Hb values (g/L) pre- and post-transfusion were 59.19±15.67 vs 77.52±15.09 in group A, 56.35±14.08 vs 74.44±15.63 in group B, 56.00±12.06 vs 75.00±4.73 in group C, respectively. The Hb values of post-transfusion for three groups were all higher than those of pre-transfusion (P<0.05). 【Conclusion】 Anti-E from Rh system is the most common antibody in patients with unexpected antibody. Appropriate red blood cells transfusion with Hb increases by an average of 6-7 g/L per 1 U of red blood cells indicating good transfusion efficacy. For positive DAT patients, transfusion of suspended red blood cell is feasible.
9.Effect and mechanism of extracorporeal photopheresis on preventing acute graft versus host disease in ice
Zhanrui CHENG ; Yuwei LIN ; Yan ZHONG ; Along ZHANG ; Haixia XU ; Li TIAN ; Zhong LIU
Chinese Journal of Blood Transfusion 2023;36(10):860-867
【Objective】 To investigate the preventive effects of early apoptotic splenic mononuclear cells induced by extracorporeal photopheresis (ECP) on acute graft versus host disease (aGVHD) in mice and explore the underlying mechanisms. 【Methods】 1) Splenic mononuclear cells were extracted from C57BL/6 mice and treated with different concentrations of 8-MOP (50 ng/mL, 100 ng/mL, 200 ng/mL, 300 ng/mL, 600 ng/mL). After treatment, irradiate the cells with 2 J/cm2 of ultraviolet light. Then, use the Annexin V-FITC/PI apoptosis detection kit to assess the early apoptosis rate of the cells and determine the optimal concentration of 8-MOP for the experiment.2) There were 35 SPF-grade female BALB/C mice (H-2Kd) aged 6-8 weeks. After whole-body irradiation with 8Gy X-rays, the mice were divided into five groups: sham irradiation group received intravenous infusion of 0.2 mL of normal saline, the syngeneic bone marrow transplantation group received intravenous infusion of 0.2 mL of BALB/C mouse bone marrow nucleated cell suspension (including a cell count of 1×107), the allogeneic bone marrow transplantation group received intravenous infusion of 0.2 mL of C57BL/6 mouse bone marrow nucleated cell suspension (including a cell count of 1×107), the aGVHD group received intravenous injection of a mixture of C57BL/6 mouse bone marrow nucleated cells (including a cell count of 1×107) and splenic mononuclear cells (including a cell count of 1×107) in 0.2 mL, the ECP prevention group received pre-transplant intravenous infusion of 0.2 mL of ECP-treated splenic mononuclear cells of C57BL/6 mice (including a cell count of 1×107 ) 48 hours before transplantation, and on the day of transplantation, intravenous injection of a mixture of C57BL/6 mouse bone marrow nucleated cells (including a cell count of 1×107) and splenic mononuclear cells (including a cell count of 1×107 ) in 0.2 mL.The preventive effects of ECP on aGVHD were observed, and the concentrations of IFN-γ, IL-2, TNF, IL-4 and IL-6 in mouse serum were measured using CBA. Th1 cell counts were determined by flow cytometry. 【Results】 Different concentrations of 8-MOP (50 ng/mL,100 ng/mL, 200 ng/mL, 300 ng/mL, 600 ng/mL) were used to treat mouse splenic mononuclear cells. The early apoptosis rates (%), observed after treatment were as follows: (14.18±0.865) vs (16.76±0.407) vs (18.83±0.404) vs (19.27±0.404) vs (14.5±0.529). The appropriate concentration of 8-MOP was determined to be 200 ng/mL.In vivo experiment, the results showed that the aGVHD group had decreased survival rate, reduced body weight, and increased clinical scores compared to the syngeneic and allogeneic bone marrow transplantation groups (P<0.01), and the chimerism of bone marrow cells in mice after transplantation was over 90%. ECP significantly improved the survival rate of mice after transplantation, reduced clinical scores (P<0.05), and decreased the concentrations of Th1 cell cytokines in serum (P<0.05) and the counts of Th1 cells in the spleen (P<0.05). 【Conclusion】 ECP-induced early apoptotic single nuclear cells from the spleen can prevent the occurrence of aGVHD by reducing the Th1 response in mouse.
10.Molecular tracking of long-distance transmission events of COVID-19 by lorry drivers in Fujian province
Qi LIN ; Zehui CHEN ; Zhimiao HUANG ; Xiaohong ZENG ; Quwen LI ; Yuwei WENG
Chinese Journal of Experimental and Clinical Virology 2023;37(1):55-60
Objective:To confirm the cross-provincial long-distance transmission events of Coronavirus Disease 2019 (COVID-19) by lorry drivers, the origin of infections of the cases and the transmission routes were tracked.Methods:Nasopharyngeal swab specimens from five lorry driver cases of COVID-19, found in Zhangzhou city in March, 2022 when the local outbreaks occurred in adjacent Quanzhou city, Fujian province, were collected to perform 2019 novel coronavirus (2019-nCoV) targeted genome amplification and followed by next-generation sequencing. The sequences were submitted to online 2019-nCoV analysis platforms to classify the type of variant and mutation sites. Phylogenetic tree for the viruses were constructed by phylogenetic analysis software. Combined with the epidemiological investigation, the origin of infections of the cases and the transmission routes were deduced.Results:Five complete genome sequences, with 29 770-29 839 bp in length and 99.53% average genomic coverage, of 2019-nCoV were successfully obtained. The viruses were all Omicron variants and further divided into three different subclades of BA.2. Of the five strains of 2019-nCoV, three were highly similar to the viruses of two distinct lineages co-circulated in Quanzhou city during the period of local outbreak of COVID-19, respectively. Phylogenetic analysis also revealed that the viruses from three infected lorry drivers were highly homologous to that from local outbreaks in Quanzhou city. The viruses from the rest two cases had seven to fourteen nucleotide mutations (corresponding to 5-7 amino acid substitutions) when compared with the viruses in local outbreaks in Quanzhou city, which excluded the involvement of the two cases into the transmission chains of local outbreaks. Combined with the field epidemiological investigations, the result revealed that the origin of infection of 2019-nCoV of the two sporadic lorry driver cases was outside of Fujian province.Conclusions:With the aid of high-throughput sequencing and bioinformatics technology combined with field epidemiological investigations, we speculated in this study that at least three origins of infection of 2019-nCoV in five lorry driver cases and cross-provincial long-distance transmission via two sporadic cases infected outside Fujian province when they returned.


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