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Exosomes are extracellular vesicles that are released from the cell into the extracellular microenvironment, and are a carrier of biological information transmitted over long distances, which have a dual regulatory effect on the immune system. Studies have shown that viruses can hold the endosomal-exosomal pathway hostage to spread. This review describes the role played by exosomes in viral infections. On the one hand, exosomes can transmit antiviral substances and trigger immune responses against viral infections, and on the other hand, exosomes can serve as potential mediators of viral transmission, which have been shown to be closely related to the transmission and immune escape of a variety of viruses. Understanding the role of exosomes in viral infections is essential for the detection of viruses in the blood, for improving blood safety, and for the search and development of antiviral drugs.
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【Objective】 To retrospectively analyze the blood use of transfusion-dependent thalassemia (TDT) patients in 9 designated transfusion medical institutions from 2018 to 2023 in Nanning, and to evaluate the effect of " three designated " blood transfusion mode (hereby means TDT patients undergoing blood transfusion in designated transfusion medical institutions regularly) and " collection-based-supply" blood management mode on blood security of TDT patients. 【Methods】 The " three designated" blood transfusion mode was implemented to ensure that TDT patients registered in the local household registration (referred to as the " register" ) obtain the rights and interests of outpatient transfusion and blood security of designated medical institutions. The " collection-based-supply" blood management mode was implemented to assess the blood needs of "register" TDT patients and meet their needs to the maximum extent according to the blood inventory (collection). 【Results】 From 2018 to 2023, the total blood supply of "register" TDT patients was 10.37% of the total red blood supply of all medical institutions (138 509.5 U /1 335 788.0 U), with the highest proportion of type O blood as 46.34% (64 181.0 U/138 509.5 U) and the lowest proportion of type AB blood as 3.85% (5 331.0 U/138 509.5 U). In 2018, 9 transfusion medical institutions were designated for TDT patients.There were a total of 766 TDT patients in the register, with the per capita annual blood transfusion volume increased from 20.28 U (15 531.0 U/766 patients) in 2018 to 36.01 U (27 586.0 U/766 patients) in 2023, maintaining a positive growth every year(30.26%, 4.94%, 11.71%, 8.61%, 4.94% and 7.10%). 【Conclusion】 The " three designated" blood transfusion mode and the " collection-based-supply " blood management mode can effectively guarantee the blood supply of TDT patients.
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In the context of Chinese narrative medicine， narrative foreclosure is an abnormal state in the process of life narrative， which is the “fractured or stagnation of the life narrative process”. Based on analyzing the characteristics and performance of typical traumatic narrative foreclosures in literary works and clinical reality， this paper proposed that narrative care is of great significance for the traumatic subject to get out of narrative foreclosures and re-enter the narrative process of mental and physical comfort. “Narrative care” is a way of care in which medical staff or educator in health institutions use their narrative capital and narrative wisdom to establish interpersonal narrative connections with themselves， their families， and service or education recipients， so as to nourish subjects in narrative foreclosure and help them get out of life’s dilemmas. Narrative caregivers assist the traumatic subjects in gaining the restorative power to repair the fractured narrative process of life through the construction of a narrative community， getting out of isolation， achieving growth， and restoring overall health.
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Cerebral organoids are three-dimensional nerve cultures induced by embryonic stem cells(ESCs)or induced pluripotent stem cells(iPSCs)that mimic the structure and function of human brain.With the continuous optimization of cerebral organoid culture technology and the combination with emerging technologies such as organ transplantation,gene editing and organoids-on-chip,complex brain tissue structures such as functional vascular structures and neural circuits have been produced,which provides new methods and ideas for studying human brain development and diseases.This article reviews the latest advances in brain organoid technology,describes its application in neurological diseases and advances in stroke modeling and transplantation treatment.
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Post-stroke cognitive impairment(PSCI)is mainly manifested as learning and memory disorders.Highly enriched RNA m6A methylation modification in mammalian brain is involved in glial cell-mediated neuroinflammation.Given that neuroinflammation is the main mechanism for neural damage and spatial and memory impairment of PSCI,it is speculated that RNA m6A methylation modification can regulate the inflammatory response of glial cells after stroke to improve PSCI.This review summarizes and analyzes the role of RNA m6A methylation modification in the development of PSCI and analyzes its detailed mechanism of regulating glial cell-mediated inflammation,which will provide reference for researchers in this field.
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BACKGROUND:Long non-coding RNAs(lncRNAs),as important regulators of the inflammatory response,are involved in the immune-inflammation-brain crosstalk mechanism after ischemic stroke and have the potential to become a therapeutic agent for neurological dysfunction after ischemic stroke. OBJECTIVE:To analyze and summarize the molecular mechanism of lncRNA acting on glial cells involved in the neuroimmuno-inflammatory cascade response after ischemic stroke and the associated signaling pathways,pointing out that lncRNAs have the potential to regulate inflammation after ischemic stroke. METHODS:PubMed was searched using the search terms of"ischemic stroke,long non-coding RNA,neuroinflammation,immune function,signal pathway,microglia,astrocytes,oligodendrocyte,mechanism,"and 63 relevant documents were finally included for review. RESULTS AND CONCLUSION:In the early stage of ischemic stroke,the death of nerve cells due to ischemia and hypoxia activates the innate immune response of the brain,promoting the secretion of inflammatory factors and inducing blood-brain barrier damage and a series of inflammatory cascades responses.As an important pathogenesis factor in ischemic stroke,the neuroimmuno-inflammatory cascade has been proved to seriously affect the prognosis of patients with ischemic stroke,and it needs to be suppressed promptly in the early stage.Neuroinflammation after ischemic stroke usually induces abnormal expression of a large number of lncRNAs that mediate a series of neuro-immune-inflammatory crosstalk mechanisms through regulating the polarization of microglia,astrocytes and oligodendrocytes to exert post-stroke neuroprotective effects.LncRNAs,as important regulatory factors of the inflammatory response,inhibit the neuroimmuno-inflammatory cascade response after ischemic stroke through regulating nuclear factor-κB,lncRNA-miRNA-mRNA axis,Rho-ROCK,MAPK,AKT,ERK and other signaling pathways to effectively improve neurological impairment after ischemic stroke.Most of experimental studies on the interaction between lncRNAs and ischemic stroke are based on a middle cerebral artery occlusion model or a cerebral ischemia-reperfusion injury model,but no clinical trials have been conducted.Therefore,it remains to be further explored about whether lncRNAs can be safely applied in clinical practice.At present,there are many therapeutic drugs for the treatment of ischemic stroke,but there are relatively few studies on the application of lncRNAs,exosomes and other transplantation technologies for the treatment of ischemic stroke using tissue engineering technology,which need to be further explored.lncRNA has become an important target for the treatment of ischemic stroke with its relative stability and high specificity.In future studies,more types of inflammatory lncRNAs that function under ischemic-hypoxia conditions should continue to be explored,in order to provide new research directions for the treatment of neuroinflammation after ischemic stroke.
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BACKGROUND:It has been hypothesized that PANoptosis may be involved in the pathologic process of osteoporosis,but there have been no studies addressing the mechanisms of PANoptosis genes in osteoporosis. OBJECTIVE:To analyze the biological mechanism of PANoptosis regulators in the occurrence and development of osteoporosis. METHODS:The GSE56815 dataset was obtained from the Gene Expression Omnibus database and PANoptosis genes were extracted for differential analysis.The key genes of PANoptosis were screened by random forest tree model to construct a disease risk prediction model.Consensus clustering algorithm,single sample genome enrichment analysis and immune infiltration analysis were used to explore the differences between different PANoptosis molecular subtypes.Herbal drugs that regulate the key genes of PANoptosis were predicted through Coremine medical database,a medical ontology information retrieval platform. RESULTS AND CONCLUSION:Based on the four PANoptosis key genes(CASP1,CASP10,MEFV,and TNF),the diagnostic markers of osteoporosis were determined,and the risk prediction model was constructed and verified.Osteoporosis was divided into two different PANoptosis subtypes(clusters A,B and gene clusters A,B),and the PANoptosis scores of cluster B and gene cluster B were higher than those of cluster A and gene cluster A,respectively.Traditional Chinese drugs such as ginseng which can regulate the key genes of PANoptosis were predicted by the Coremine medical database.
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Objective To observe the effects of electroacupuncture on the motor function and mitochondrial dynamics of skeletal muscle of SAMP8 mice;To explore the mechanism of electroacupuncture in improving the motor dysfunction of Alzheimer disease(AD)from the perspective of mitochondrial dynamics.Methods Totally 18 SAMP8 mice were divided into model group and electroacupuncture group,with 9 mice in each group,and the SAMR1 mice with the same age were set as control group."Baihui","Dazhui"and"Shenshu"were selected in the electroacupuncture group,and electroacupuncture was performed daily for 20 min,8 d as a course of treatment.Each course of treatment was separated by 2 d,for a total of 3 courses of treatment.The model group and the control group were not intervened.The motor function of mice was tested by grip strength test,suspension test,hind limb extension test and Morris water maze experiment.The morphology and structure of gastrocnemius were observed by HE staining,ATP content in gastrocnemius was determined by colorimetry,the mRNA expression of optic atrophy 1(OPA1),mitofusin 2(MFN2)and dynamin-related protein 1(DRP1)in gastrocnemius were detected by real-time quantitative PCR,the expressions of OPA1,MFN2 and DRP1 in gastrocnemius were detected by Western blot.Results Compared with the control group,the grip strength,the score in suspension test,and the average speed and maximum speed of Morris water maze experiment of mice in model group significantly decreased(P<0.01);the arrangement of fibers in the gastrocnemius muscle tissue was disordered,the gaps become wider,and the distribution of nuclei was uneven;the ATP content in the gastrocnemius muscle tissue was significantly decreased(P<0.01),the mRNA and protein expressions of OPA1 and MFN2 were significantly decreased(P<0.01),and the expression of DRP1 mRNA and protein significantly increased(P<0.01).Compared with the model group,the grip strength,the score in suspension test,and the average speed and maximum speed of Morris water maze experiment in electroacupuncture group significantly increased(P<0.01);the arrangement of gastrocnemius muscle tissue was relatively neat,the gaps become narrower,and the distribution of nuclei was more uniform;the ATP content in gastrocnemius muscle tissue significantly increased(P<0.01),while the mRNA and protein expressions of OPA1 and MFN2 significantly increased(P<0.05,P<0.01),the expression of DRP1 mRNA and protein significantly decreased(P<0.01).Conclusion Electroacupuncture can improve the skeletal muscle morphological structure and motor dysfunction of SAMP8 mice,and the mechanism may be related to the correction of skeletal muscle mitochondrial dynamic imbalance and the increase of skeletal muscle ATP content.
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At present, many infectious pathogens, especially emerging/re-emerging pathogens, exist in the blood of voluntary blood donors and may be transmitted through blood transfusions. However, most of Chinese blood centers only routinely screen for HBV, HCV, HIV, and syphilis. We employed metagenomic next-generation sequencing (mNGS) to investigate the microbiome in healthy voluntary blood donors to help assess blood safety in China by identifying infectious pathogens presented in donations that could lead to transfusion-acquired infections. We collected 10,720 plasma samples from voluntary blood donors from seven blood centers in different cities during 2012-2018 in China. A total of 562 GB of clean data was obtained. By analyzing the sequencing data, it was found that the most commonly identified bacteria found in the healthy blood were Serratia spp. (5.0176%), Pseudomonas spp. (0.6637%), and Burkholderia spp. (0.5544%). The principal eukaryote were Leishmania spp (1.3723%), Toxoplasma gondii (0.6352%), and Candida dubliniensis (0.1848%). Among viruses, Human Parvovirus B19 (B19V) accounts for the highest proportion (0.1490%), followed by Torque teno midi virus (0.0032%) and Torque teno virus (0.0015%). Since that B19V is a non-negligible threat to blood safety, we evaluated the positive samples for B19V tested by mNGS using quantitative polymerase chain reaction, Sanger sequencing, and phylogenetic analysis to achieve a better understanding of B19V in Chinese blood donors. Subsequently, 9 (0.07%) donations were positive for B19V DNA. The quantitative DNA levels ranged from 5.58 × 102 to 7.24 × 104 IU/ml. The phylogenic analyses showed that prevalent genotypes belonged to the B19-1A subtype, which disclosed previously unknown regional variability in the B19V positivity rate. The investigation revealed that many microbes dwell in the blood of healthy donors, including some pathogens that may be dormant in the blood and only cause disease under specific conditions. Thus, investigating the range and nature of potential pathogens in the qualified donations provided a framework for targeted interventions to help prevent emerging and re-emerging infectious diseases.
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Objective:To explore the application effect of case-based learning (CBL), teaching mode combined with 3D printing in clinical teaching of sacral tumors.Methods:A total of 108 undergraduate interns and standardized residency training students who studied in our hospital from 2017 to 2018 were divided into the CBL teaching group ( n = 53) and the CBL combined with 3D printing teaching group ( n = 55) according to their study time. The combined teaching group used computer tomography (CT) data to reconstruct and print out a 3D model of sacral tumors based on CBL, and performed preoperative teaching on the invasion of the surrounding tissues of the tumor. The scores of the students in the two groups were evaluated respectively, and the students were surveyed by self-identification questionnaire (learning interest, self-learning ability, teamwork ability, comprehensive analysis ability and clinical thinking ability). The t-test (one-sided) was used for comparison between groups using stata 14.0. Results:The score of CBL teaching group (75.90±6.70) was lower than that of CBL combined with 3D printing teaching group (83.60±7.40). In terms of critical thinking ability evaluation, self-learning ability, learning interest, comprehensive analysis ability and clinical thinking ability, the CBL combined 3D printing teaching group was superior to the CBL teaching group, and the difference was statistically significant ( P<0.001). In terms of teamwork ability, there was no statistical difference between the two groups. Conclusion:The CBL teaching mode combined with 3D printing can improve academic performance, students' learning interest and clinical thinking ability of sacral tumors in the teaching of undergraduate interns and standardized residency training students.
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【Objective】 To monitor the positive rates of IgM and IgG antibodies and the changes in S/CO values over time in voluntary blood donors infected with SARS-CoV-2 after recovery, in order to provide theoretical basis and data support for ensuring clinical blood safety. 【Methods】 A total of 54 platelet donors who met the inclusion criteria were selected for this study, and 359 blood samples (plasma) from T1 to T7 (at 7 time points, spanning 14 to 15 weeks) were continuously monitored for SARS-CoV-2 IgM and IgG antibodies using fully automated chemi-luminescence method. 【Results】 Among 359 blood samples (plasma) from 54 donors, 349 were with valid test results.Eleven donors were tested positive for IgM, with a positive rate of 20.37%, and IgM antibody S/CO value gradually increased during T1-T3, but gradually decreased during T4-T7. Fifty-four donors were tested positive for IgG, with the positive rate of 100%, and the S/CO value of IgG antibody gradually decreased over time. During the period of T1 -T7, there was no significant difference in SARS-CoV-2 IgG antibody S/CO value between gender (male/female) or age group (18-39 years old /40-60 years old). 【Conclusion】 The positive rate of SARS-CoV-2 IgG antibody in platelet donors after recovery from SARS-CoV-2 infection was 100% during 2-14 weeks, and the IgG S/CO value gradually decreases over time. The long-term dynamic changes of SARS-CoV-2 serologically specific IgG and IgM antibodies in blood donors are worthy of further study.
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Objective:To analyze the effect of spine-pelvis sagittal parameters and sagittal orientation of facet joint on degeneration of cranial L 3,4 facet joint (facet joint degeneration, FJD) after L 4-S 1 posterior lumbar interbody fusion (PLIF). Methods:Patients with lumbar degenerative diseases who underwent L 4-S 1 PLIF from January 2012 to December 2016 were retrospectively investigated, there were 54 cases, including 28 males and 26 females. Age: 54.59±5.48 years (range, 45-60 years). X-ray, CT, MRI and Weishuapt grade was used to evaluate the degeneration of L 3,4 facet joint at the cranial adjacent segment. The general information and the sagittal parameters of spine pelvis at the last follow-up were compared between the two groups. The former included age, gender, body mass index (BMI), bone mineral density (BMD), follow-up time and preoperative diagnosis. The latter included lower lumbar lordosis angle (LLL), lumbar lordosis angle (LL), pelvis incidence (PI), pelvis tilt (PT), sacrum slope (SS), the height of the intervertebral space (HD), the angle of cranial facet joint, Oswestry disability index (ODI), Japanese Orthopedic Association (JOA) lumbar function score and improvement rate were compared at the same time. Independent sample t-test was used to compare continuous variables between groups; comparison of categorical variable components χ 2 test or Fisher's exact test. Multivariate logistic regression analysis was used to predict the risk factors of adjacent FJD. Results:Postoperative follow-up was 33.44±6.85 months (range, 24-36 months), there were 17 patients in the degenerative group and 37 patients in the non degenerative group. There were no significant differences in age, gender, BMI, BMD, follow-up time or preoperative diagnosis between the two groups. LLL, LL and SS also showed no significant difference. At the last follow-up, PI (56.28°±6.03° vs. 47.87°±8.30°, t=3.74, P=0.001), PT (17.90°±7.06° vs. 14.41°±5.51°, t=1.97, P=0.042) and the joint angle of the cephalic facet (58.48°±2.00° vs. 54.69°±3.01°, t=4.72, P=0.072) in the degenerative group were greater than those in the non-degenerative group. In the subgroup analysis of lumbar lordosis distribution, the difference between the two groups was statistically significant (χ 2=9.90, P=0.006). The HD in the degenerative group 7.50±3.60 mm was significantly lower than that in the non degenerative group 9.30±2.79 mm ( t=2.00, P=0.031). Multivariate logistic regression analysis showed that increase of PI ( OR=1.22, P=0.005) and magnified cephalic facet joint angle ( OR=2.04, P=0.008) were risk factors for adjacent segment facet degeneration. At the last follow-up, the ODI improvement rate in the degenerative group (58.14%±13.41% vs. 70.18%±8.03%, t=4.11, P<0.001) and the JOA score improvement rate (44.72%±9.53% vs. 68.86%±8.55%, t=0.43, P=0.001) were lower than those in the non degenerative group. Conclusion:The increase of PI and sagittal facet (increased joint angle of proximal facet) are risk factors of adjacent segment FJD after lumbar fusion; The abnormal distribution of lower lumbar lordosis and poor PT recovery in adjacent segment FJD patients after lumbar fusion are more obvious, which may be related to the increase of PI; After lumbar fusion, the orientation of adjacent facet joint tended to be sagittal.
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Objective:To investigate the safety and efficacy of nimotuzumab combined with IMRT radiotherapy in the treatment of elderly patients with locally advanced cervical cancer (LACC).Methods:A retrospective analysis was conducted for 34 elderly LACC patients treated with nimotuzumab combined with IMRT radiotherapy or concurrent chemoradiotherapy in the Zhangzhou Affiliated Hospital of Fujian Medical University from June 2020 to December 2021. The efficacy and side effects were evaluated one and two years after treatment.Results:Median follow-up time was 13.3 months (6.1-24.3 months). A total of 24 cases of complete response (CR) and eight cases of partial response (PR) were achieved after treatment, with an objective response rate (ORR) of 94.1% (32/34). The tumor diameters were (49.56 ± 19.22) mm before treatment and (19.61 ± 14.59) mm after treatment, with a tumor regression rate (TRR) of 59.22%.The 1- and 2- year progression-free survival (PFS) rates were 84.9% and 84.9%, overall survival (OS) rates 91.8% and 87.2%, respectively, disease-free survival (DFS) rates 91.8% and 87.2%, respectively, and the cancer-specific survival (CSS) rates 95.7% and 90.9%, respectively. The main adverse events included radiation enteritis, leukopenia, hypoproteinemia and anemia.Conclusions:Nimotuzumab combined with IMRT radiotherapy or concurrent chemoradiotherapy is safe and effective in the treatment of LACC.
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【Objective】 To assess three severe acute respiratorysyndrome coronavirus 2 (SARS-CoV-2) enzyme linked immunosorbent assays (ELISA) and one pseudotype lentivirus-based neutralization test (ppNAT) in detecting the convalescent plasma antibody levles from COVID-19. 【Methods】 30 COVID-19 convalescent plasma samples were screened for antibodies against SARS-CoV-2 using three kinds of SARS-CoV-2 ELISA reagents and one ppNAT test in Shenzhen. The controls consisted of plasma samples from 32 healthy blood donors in February 2019. The diagnostic efficacy analysis of various SARS-CoV-2 ELISA reagents was performed using real-time fluorescent Polymerase Chain Reaction (RT-PCR). We also analyzed correlation between different immunological reagents and the age, gender, hospitalization, and severity of illness. 【Results】 The positive yielding rate of ppNAT and three kinds of IgG ELISA was higher than that of IgM ELISA. The positive yielding rates of three kinds of IgG ELISA were 100%(30/30), 93.33%(28/30), and 96.67%(29/30) respectively, while the yielding rates in control group were all 0. The positive yielding rate of three IgM ELISAs were 93.33%(28/30), 70%(21/30)and 46.67% (14/30). All the cases from negative control group were negative for IgG and IgM. Pearson correlation coefficient was calculated; there was a strong correlation between ELISA reagent 2 IgG and ELISA reagent 3 IgG (r=0.765, P0.05). 【Conclusion】 In the convalescent plasma with nucleic acid confirmed covid-19, the yielding rates of different IgM antibodies varied greatly. Antibody levels were influenced by age to some extent.
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【Objective】 To investigate the distribution of Hepatitis B virus(HBV)genotypes and the genetic characteristics of genotype B HBV populations among voluntary blood donors in five regions of China. 【Methods】 A total number of 630 plasma samples from blood donors with positive HBV HBsAg neutralization test from 2014 to 2016 in Guangxi Blood Center, Chongqing Blood Center, Urumqi Blood Center, Mianyang Central Blood Station and Luoyang Central Blood Station were collected. The S-region sequence of the HBV genome was amplified by semi-nested PCR and followed with Sanger sequencing in order to investigate the HBV genotype distribution and population genetics. 【Results】 Among the voluntary blood donors in five regions, 55 cases of HBV S gene fragments were successfully amplified. Three genotypes were found in HBV typing, including 46 cases of type B(83.64%), 7 cases of type C(12.73%) and 2 cases of type D(3.63%). There were 15 cases of type B and 3 cases of type C in Guangxi; 10 cases of type B and 1 case of type C in Chongqing; 3 cases of type B, 1 case of type C and 1 case of type D in Luoyang; 15 cases of type B in Mianyang; 3 cases of type B, 2 cases of type C and 1 case of type D in Urumqi. The mismatch distribution of the HBVB type population with the largest population number showed a unimodal distribution, and the results of Tajima′s D test and Fu′s Fs test were both negative, indicating that the HBV population in these five regions was expanding. 【Conclusion】 The prevalent genotype of HBV in voluntary blood donors is type B, and the type B HBV population is experiencing a slow expansion, which should attract our attention. In the future, a thorough molecular epidemiological investigation of HBV should be carried out to ensure blood safety.
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【Objective】 To investigate the effect of exosomes produced by hepatitis B virus (HBV)-infected cells on the phenotype and function of macrophages. 【Methods】 The exosomes secreted by HepAD38 cells, which were capable of producing HBV and HepG2 cells, were collected by ultracentrifugation combined with immunosorbent method.The quality and purity of the extracted exosomes were verified by nanoparticle tracking analysis (NTA), scanning electron microscope and Western blot.The M0 THP-1 macrophages differentiated by PMA were stimulated by HepAD38 derived- or HepG2 derived exosomes.Total RNA and protein samples were collected at different time points after stimulation.Real-time fluorescence quantitative PCR (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect cytokine mRNA and protein expressions, respectively.Meanwhile, neutral red assay was performed to analyze macrophage pinocytosis activity, and a commercial kit was used to measure reactive oxygen species (ROS) in THP-1 macrophages.Human reverse transcription chip detection was performed to obtain the microRNAs profile of the exosomes.And the effect of selected miRNA on macrophages was further confirmed by qRT-PCR. 【Results】 Compared with HepG2-derived exosomes, HepAD38-derived exosomes increased the mRNA and protein expressions of IL-1β, MCP-1 and TNFα significantly.However, no difference of pinocytosis capacity or ROS production was found between the HepAD38-derived exosomes group and HepG2-derived exosomes group.Human reverse transcription chip detection results were verified by KEGG analysis and qRT-PCR, and it was found that miR-6824-3p could also significantly increase the expression levels of IL-1β, MCP-1 and TNFα after high expression. 【Conclusion】 This study found that exosomes produced by HepAD38 cells may stimulate macrophages to produce inflammatory factors such as IL-1β, MCP-1 and TNFα through miR-6824-3p, thereby playing a role in HBV infection.
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【Objective】 To analyze the metagenomics and microbiology of voluntary blood donors in China, so as to assess the potential threats of emerging infectious diseases to the safety of blood transfusion. 【Methods】 12 300 plasma samples (10 mL each) collected by central blood stations in Chongqing, Liuzhou, Urumqi, Mianyang, Wuhan, Nanjing, Mudanjiang, and Dehong Prefecture area from 2012 to 2018 were subjected to total DNA extraction after ultracentrifugation (32 000 rpm/min, centrifugal radius 91.9 mm) in minipools of 160 donations. The metagenomic library was constructed, and deep sequencing was conducted by Illumina Hiseq 4 500. By comparing with reference sequences of bacteria, fungi, parasites and viruses, metagenomic data were analyzed, classification of microbes were identified, and potentially harmful pathogens were evaluated. 【Results】 A total of 632 GB clean data were obtained by deep sequencing, and the top three pathogens were Pseudomonas(0.561 1%), Burkholderia(0.468 7%) and Serratia(4.242 0%). Pathogens with potential threat which could be transmitted by blood transfusion or blood products were found, such as human parvovirus B19(0.126 6%), Leishmania spp(1.348 5%) and Toxoplasma gondii(0.615 8%). 【Conclusion】 Our study analyzed metagenomics of voluntary blood donors in parts of China and revealed pathogens that may cause potential harm to blood safety, which were helpful for targeted prevention and control of emerging infectious diseases.
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【Objective】 To map a comprehensive description of microbiome presented in healthy blood donations in Liangshan Yi Autonomous Prefecture, and detect the potential pathogens and its prevalence. 【Methods】 A total of 1 299 blood samples were randomly selected from healthy blood donors in Liangshan Prefecture. Total nucleic acids were extracted and sequenced by high-throughput sequencing, and the microbiome was determined by metagenomics analysis. Quantitative real-time PCR (qPCR) and ELISA assays were used to detect antibodies against HSV DNA and HSA in each sample. The prevalence of HSV in healthy blood donors was compared in terms of gender, age, occupation, education level, and frequency of donation. 【Results】 3.49GB data were obtained from DNA pool through high-throughput sequencing. After filtering the data of human genome, the DNA sequences annotated as bacteria, parasites / fungi and viruses were 213 057, 10 623 and 15 reads, respectively. A total of 2.79GB data were obtained from cDNA pool, after filtering the data of human genome, the fragments annotated as bacteria, parasites / fungi and viruses were 4 105 600, 18 446 and 0 reads, respectively. The prevalence of IgG and / or IgM antibodies against herpes simplex virus type 1 and 2 were 8.62% (112 / 1 299) and 18.32% (238/1 299), and that of nucleic acid was 0.77 ‰ (1/1 299). 【Conclusion】 The microbiome of healthy blood donors in Liangshan Prefecture and the potential pathogens were identified in this study. Regional specificity of HSV infections emerged in Liangshan Prefecture. Considering the collaboration between HSV-2 and HIV infection, epidemiological investigation of HSV-2 infection should be conducted preferentially among different populations in Liangshan Prefecture and other HIV high prevalence areas in order to benefit the prevention and control of HIV.
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【Objective】 To investigate the effect of transfusion-transmitted Zika virus (ZIKV) on the expression of non-coding circular RNA (hsa_circ_0001613) and the role of hsa_circ_0001613 in Zika virus replication. 【Methods】 Human adenocarcinomic alveolar basal epithelial cells (A549) were seeded on a 12-well plate at 1.8×105/ well and infected with ZIKV at 0.05 MOI. The Total RNAs were isolated every day for 5 days after infection, and the relative expression level of hsa_circ_0001613 was detected by qRT-PCR. In addition, 10nM siRNA-hsa_circ_0001613 was transfected into 2×105/ well A549 cells to specifically knock down the expression level of hsa_circ_0001613. 24h later, the cells were infected with ZIKV (MOI=0.05). Total RNAs were isolated at day 1-5 post-infection, proteins were extracted 96h post-infection. ZIKV replication, relative host antiviral gene expression, and interferon stimulated response element (ISRE) activity were tested using qRT-PCR, western blot and dual luciferase assay, respectively. 【Results】 The relative expression of hsa_circ_0001613 decreased significantly after 1-5 days of ZIKV infection. Knockdown of hsa_circ_0001613 inhibited ZIKV replication. Meanwhile, hsa_circ_0001613 knockdown significantly upregulated IFN-α/β and its downstream interferon-stimulated genes (ISGs) expression, also increased ISRE activity. 【Conclusion】 ZIKV infection significantly suppressed hsa_circ_0001613 expression in A4549 cells. Preliminary study indicated that hsa_circ_0001613 knockdown inhibited ZIKV replication possibly through activating type-Ⅰ IFN signaling pathway as showed by increased ISGs expression and ISRE activity.
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【Objective】 To investigate the role of non-coding microRNA miR-16-5p in ZIKV replication and the underlying mechanism. 【Methods】 1×105/mL HeLa cells were seeded in 24-well plate and infected with ZIKV(MOI=5). RNAs were harvested, and miR-16-5p expression levels were measured by qRT-PCR at 24, 48 and 72 hour post infection, respectively. HeLa cells were transfected with 20nM miR-16-5p mimic and infected with ZIKV(MOI=5) at 24h post transfection. RNAs were extracted and ZIKV RNA and several inflammation factors expression were tested using qRT-PCR at 48h post infection. HeLa cells were co-transfected with 1μg NFκB-luc and 10ng pRL-TK with 20nM miR-16-5p mimic, and then infected with ZIKV(MOI=5) for 24h before the luciferase expression was tested at 48h post infection. HeLa cells were transfected with 20nM miR-16-5p mimic and infected with ZIKV(MOI=5) at 24h post transfection, and cell apoptosis was assayed through flow cytometry. 【Results】 Compared with uninfected control, miR-16-5p expression was significantly decreased at 24h, 48h and 72h following ZIKV infection. MiR-16-5p over-expression inhibited ZIKV replication, while upregulated NFκB activity and inflammation factors expression compared with the negative mimic-transfected cells. MiR-16-5p overexpression also promoted HeLa cell apoptosis. 【Conclusion】 ZIKV infection downregulated intracellular miR-16-5p expression. Overexpression of miR-16-5p suppressed ZIKV infection. MiR-16-5p inhibited ZIKV replication and promoted cell apoptosis probably by activating NFκB pathway and stimulating inflammation factors expression.