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ObjectiveTo explore the clinical features and causative genes of short stature children with unknown etiology， providing evidence for precise clinical diagnosis and treatment. MethodsThe study recruited children with suspected but undiagnosed short stature from the pediatric endocrinology department in our hospital between January 2018 and August 2022. A retrospective analysis was performed on the clinical manifestations， laboratory test and whole exome sequencing （WES） results. Causative genes were classified and analyzed according to different pathogenic mechanisms. ResultsA total of 48 children （30 boys and 18 girls） were enrolled， aged 7.73 ± 3.97 years， with a height standard deviation score （ HtSDS） of -3.63 ± 1.67. Of the patients， 33 （68.8%） suffered from facial anomalies， 31 （64.6%） from skeletal abnormalities， 26 ［54.2%， 61.5% of whom born small for gestational age （SGA）］ from perinatal abnormalities， 24 ［50.0%， 87.5% of whom with growth hormone （GH） peak concentration below normal］ from endocrine disorders and 21（43.8%） had a family history of short stature. Laboratory tests showed that GH peak concentration following stimulation test was （9.72 ± 7.25） ng/mL， IGF-1 standard deviation score was -0.82 ± 1.42， the difference between bone age and chronological age was -0.93 ± 1.39 years. Of the 25 cases with mutant genes found by WES， 14 （56.0%） had pathogenic mutation， 6 （24.0%） likely pathogenic mutation， and 5 （20.0%） mutation of uncertain significance. Pathogenic and likely pathogenic variants were identified in 14 genes， including 10 affecting intracellular signaling pathways （PTPN11， RAF1， RIT1， ARID1B， ANKRD11， CSNK2A1， SRCAP， CUL7， SMAD4 and FAM111A） and 4 affecting extracellular matrix （ECM） components or functions （ACAN， FBN1， COL10A1 and COMP）. ConclusionsA rare monogenic disease should be considered as the possible etiology for children with severe short stature accompanied by facial anomalies， disproportionate body types， skeletal abnormalities， SGA， GH peak concentration below normal and a family history of short stature. WES played an important role in identifying the monogenic causes of short stature. This study indicated that affecting growth plate cartilage formation through intracellular signaling pathways and ECM components or functions was the main mechanism of causative genes leading to severe short stature in children. Further research may help discover and study new pathogenic variants and gene functions.
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【Objective】 To retrospectively analyze the serological and nucleic acid testing(NAT) data of voluntary blood donors from six blood banks in Tibet, in order to explore the positive impact of NAT on reducing the risk of infective transfusion in a regional scope. 【Methods】 From 2018 to 2022, 38 718 voluntary blood donors from blood centers of Tibet, Shannan, Shigatse, Naqu, Nyingchi and Ngari were tested for hepatitis B virus surface antigen (HBsAg), hepatitis C virus antibody (anti-HCV), human immunodeficiency virus antigen (HIV) and antibody (Ag/Ab1+2) serological determination by enzyme-linked immunosorbent assay (ELISA). At the same time, Haoyuan and Daan nucleic acid detection systems were used for the combined detection of HBV-DNA, HCV-RNA and HIV-RNA. The results of NAT of reactive ELISA samples were statistically analyzed. 【Results】 A total of 178 ELISA-/NAT+ samples were detected in Tibet over the past five years, including 170 HBV-DNA positive cases, 8 HCV-RNA positive cases, and 0 HIV-RNA positive cases, with the positive rate at 0.460%.The detection rate of 624 ELISA+/NAT+ samples was 1.61%.The age of blood donors with hepatitis B in Shigatse area was slightly higher than that in other areas, and the difference was statistically significant(P<0.05) . 【Conclusion】 The centralized detection of viral nucleic acid in Blood Center of Tibet Autonomous Region can effectively reduce the missed detection of transfusion transmitted diseases and guarantee the blood safety in the region.
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【Objective】 To explore the research status, hot spots and trends of platelet rich plasma (PRP). 【Methods】 With "platelet rich plasma (PRP)" and its Chinese equivalent as the subject words, the PRP related articles during January 1, 2013 to December 31, 2022 from PubMed and CNKI database were retrieved. The bibliometric analysis was performed by Bicomb 2.0 software to extract the annual number of literature publications, authors, journals and high-frequency theme words/sub theme words. The gCLUTO software was used to evaluate and visualize the results, and strategic diagram was drawn according to the results of biclustering. 【Results】 A total of 9 066 PRP related articles were retrieved (7 027 from PubMed, 2 039 from CNKI), and the number of publications showed an increasing trend year by year. Papers have been published in 1 527 journals in PubMed, among which the journal with the highest number of publications was Arthroscopy: Journal of Arthroscopic & Related Surgery (175 articles), followed by American Journal of Sports Medicine (171 articles ) and Journal of Cosmetic Dermatology (121 articles) . PRP-related studies were published in 541 journals in CNKI, with the top 3 journals as Chinese Tissue Engineering Research (113 articles), Chinese Aesthetic Medicine (64 articles) and Chinese Journal of Blood Transfasion (45 articles) . In Pubmed, Anitua Eduardo (84 articles), Filardo Giuseppe (53 articles) and Cole Brian J (44 articles) were the top three productive authors on PRP; Cheng Biao from the General Hospital of Southern Theater Command of Chinese PLA was the most productive Chinese author (25 articles) . Shan Guiqiu from the General Hospital of the Southern Theater Command of the Chinese People's Liberation Army published the most articles(29 articles) in CNKI. American journals published the most articles (2 745 articles ), accounting for 39.06% of the total articles, followed by British and Swiss journals, with 1 499 articles and 550 articles, respectively. A total of 42 high-frequency subject words/sub-subject words were selected from PubMed, and were classified into 6 roups, while 30 high-frequency subject words were selected from CNKI and grouped into 5 categories. The strategic coordinates show that the treatment of rotator cuff and tendon injury with PRP in PubMed, the study of PRP and tissue engineering materials in CNKI are the core themes of current PRP research. 【Conclusion】 The strategic coordinate map and bibliometrics can reveal the current research status of PRP and predict future research hotspots, but current research cores of PubMed and CNKI are not consistent, and further research is still needed.
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Objective To compare the efficacy of common clinical interventions in the treatment of cervical high-risk (HR) HPV infection based on Bayesian network meta-analysis. Methods Randomized controlled trials (RCTs) about common clinical interventions for cervical HR-HPV infection were searched in PubMed, Web of Science, Embase, The Cochrane Library, CBM, CNKI, Wanfang Data, and VIP databases from inception to July 31, 2021 using specific inclusion and exclusion criteria. The quality of the included studies was evaluated in accordance with the Cochrane systematic review manual. Meta-analysis was performed with Stata16 and RevMan5.3 software. Results Seventy-three RCTs were included, involving 3642 patients and eight treatment methods. Network meta-analysis showed that in the three months after treatment, the negative conversion rate was in the order: PTL > anti-HPV BPD > ALA-PDT > Nr-CWS > BFKS > CSJZS > rhIFNα-2b > FUO. In the six months after treatment, the negative conversion rate was in the order: Nr-CWS > ALA-PDT > PTL > anti-HPV BPD > BFKS > rhIFNα-2b > FUO > CSJZS. In the nine months after treatment, the negative conversion rate was in the order: PTL > ALA-PDT > BFKS > anti-HPV BPD > rhIFNα-2b > FUO. IN the 12 months after treatment, the negative conversion rate was in the order: Nr-CWS > ALA-PDT > anti-HPV BPD > PTL > BFKS > rhIFNα-2b > FUO > CSJZS. Conclusion In terms of HPV negative conversion rate, Nr-CWS and PTL are more effective and currently ideal compared with the other treatments. Owing to the quality of the evidence, the above conclusions must be confirmed by future high-quality studies.
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Objective:To construct a training program to improve the psychological nursing ability of clinical nurses, so as to provide a strong guarantee for the clinical development of psychological nursing.Methods:By consulting the literature, related books and investigating the curriculum of nursing colleges, the first draft of the training program was drawn up. Four departments of Cardiovascular Medicine of the Second Xiangya Hospital of Central South University from May to August 2019 were recruited and 64 nurses were trained. After the training, the training program was revised again. After two rounds of training and modification, the second draft of the training program was formed, and then Delphi method was used to conduct two rounds of expert consultation on the second draft of the training program.Results:The positive coefficients of experts in the two rounds of consultation were 94.1% and 96.7% respectively, and the average authority coefficient of experts was 0.81. The final training contents included 5 first-class indexes, 18 second-class indexes and 45 third-class indexes. The coefficient of variation of each item of training contents was 0.06-0.23, and the coefficient of variation of training methods and training duration of each part was 0.06-0.17.Conclusions:The training program is scientific, reasonable, detailed and practical, which can provide guarantee for improving the psychological nursing ability of clinical nurses.
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Objective:To investigate the effects of thioredoxin reductase 1(TR1) overexpression on hippocampus in ovariectomized SD rats.Methods:Totally 54 female SD rats were divided into normal group, ovariectomized group and ovariectomized over-expressioned TR1 group (ovariectomy-TR1 group) according to the random number table method with 18 in each group. The overexpressed TR1 vector was constructed by lentivirus, and the recombinant lentivirus was injected into the hippocampus by a brain stereotactic instrument.The mRNA levels and protein levels of TR1, Bcl-2, p53 and p21 in the hippocampus of SD rats were detected by qRT-PCR and Western blot.The expression of Caspase-3 in the hippocampus of SD rats was detected by Western blot. The activity of SOD was measured by the WST-1 cell proliferation and cytotoxicity method, the content of GSH was measured by the microplate method, and the content of MDA in the hippocampus of SD rats was measured by the TBA method. The behavioral changes of SD rats were detected by the open field test and water maze test. GraphPad Prism 9.0 was used for statistical analysis.One-way analysis of variance was used for comparisons among the three groups, and the LSD test was used for further pairwise comparisons, the t-test was used to compare the mean number of samples between the two groups. Results:(1) The mRNA and protein levels of TR1 in hippocampus of ovariectomized rats were lower than those of normal rats ( t=3.125, 4.402, both P<0.05). The mRNA and protein levels of TR1 in hippocampus of ovariectomized-TR1 rats were higher than those of ovariectomized rats ( t=4.945, 4.845, both P<0.05). (2) There were significant differences among the three groups in the escape latency in water maze test, the movement distance and the stay time in central area in the open field test ( F=44.73, 33.67, 6.51, all P<0.05), the movement distance in the open field test of ovariectomized rats was more than that of the normal group ((4 700±141) mm, (3 967±163) mm, P<0.05), the stay time in the central area was longer than that of the normal group ((87.33±3.93) s, (80.83±2.48) s, P<0.05), the movement distance ((4 267±150) mm) and the stay time in the central area ((82.17±3.43) s) of ovariectomized-TR1 group were lower than that of ovariectomized group ( P<0.05). In the water maze test, the escape latency of ovariectomized rats was longer than that of the normal group ((28.67±2.50) s, (19.50±2.59) s, P<0.05), and the escape latency in the ovariectomy-TR1 group((25.00±1.67) s) was shorter than that of ovariectomized TR1 group ( P<0.05). (3)There were significant differences in the levels of MDA, SOD and GSH in the hippocampus oxidative stress injury indexes among the three groups ( F=87.41, 91.38, 28.69, all P<0.01). The level of MDA in hippocampus of ovariectomized group was higher than that of normal group, and that in the ovariectomized-TR1 group was lower than that of ovariectomized rats group ( P<0.05). And what's more the levels of SOD and GSH in ovariectomized group were lower than those of normal group ( P<0.05), and the ovariectomized-TR1 group was higher than that of ovariectomized group ( P<0.05). (4) The results of Western blot and RT-PCR showed that the levels of p21 and p53 in hippocampal tissue of ovariectomized group were higher than those of normal group ( P<0.05), while the level of aging-related protein p21 and p53 in ovariectomized-TR1 group were significantly lower than those in ovariectomized group ( P<0.05). The level of apoptotic protein Caspase-3 in the hippocampus of ovariectomized rats was higher than that in the normal group ( P<0.05), while the level of Caspase-3 in ovariectomized-TR1 group was significantly lower than that in ovariectomized rats ( P<0.05). The level of anti-apoptotic protein Bcl-2 in hippocampus of ovariectomized group was lower than that of normal rats ( P<0.05), while the level of Bcl-2 in ovariectomized-TR1 group was significantly higher than that in ovariectomized rats ( P<0.05). Conclusion:Overexpression of TR1 can reduce apoptosis of hippocampal cells by regulating oxidative damage and reducing cell senescence.
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Objective:To construct a TPC-1 cell model that stably knocks out the HMGA2 by using CRISPR/Cas9 gene editing technology. Methods:Recombinant pLV[2gRNA]-EGFP:T2A:Puro- U6> {hHMGA2 [gRNA# A1]*}- U6>{hHMGA2 [gRNA#A2]*} of lentiviral plasmid vector was constructed: targeting HMGA2 Dual-gRNA sequence was designed, the synthesized Dual-gRNA fragment into pLV [2gRNA]-EGFP was cloned: T2A:Puro-U6 vector, extract a single clone for sequencing verification. the constructed recombinant plasmid vector with lentivirus was packed, and TPC-1 cells were infected, puromycin was used to obtain HMGA2 knock-out single clone, PCR and sequencing verification were performed, and real-time fluorescent quantitative qPCR was used to detect HMGA2 mRNA in cells Knockout efficiency. Results:After sequencing verification, pLV [2gRNA]-EGFP targeting HMGA2: T2A: Puro-U6>{hHMGA2 [gRNA#A1]*}-U6>{hHMGA2 [gRNA #A2]*} plasmid was successfully constructed; A single clone was picked for PCR identification and gene sequencing, TPC-1 cells were successfully obtained with HMGA2 gene completely knocked out; TPC-1 cells with HMGA2 knocked out were detected by real-time fluorescent quantitative qPCR, and they did not express HMGA2 mRNA.Conclusion:CRISPR/Cas9 gene editing technology enables us to construct a human papillary thyroid cancer cell line TPC-1 cell model with stable knockout of HMGA2.
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As a sensor of cytosolic DNA, the role of cyclic GMP-AMP synthase (cGAS) in innate immune response is well established, yet how its functions in different biological conditions remain to be elucidated. Here, we identify cGAS as an essential regulator in inhibiting mitotic DNA double-strand break (DSB) repair and protecting short telomeres from end-to-end fusion independent of the canonical cGAS-STING pathway. cGAS associates with telomeric/subtelomeric DNA during mitosis when TRF1/TRF2/POT1 are deficient on telomeres. Depletion of cGAS leads to mitotic chromosome end-to-end fusions predominantly occurring between short telomeres. Mechanistically, cGAS interacts with CDK1 and positions them to chromosome ends. Thus, CDK1 inhibits mitotic non-homologous end joining (NHEJ) by blocking the recruitment of RNF8. cGAS-deficient human primary cells are defective in entering replicative senescence and display chromosome end-to-end fusions, genome instability and prolonged growth arrest. Altogether, cGAS safeguards genome stability by controlling mitotic DSB repair to inhibit mitotic chromosome end-to-end fusions, thus facilitating replicative senescence.
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Inhibitor of nuclear factor kappa-B kinase subunit beta (IKKβ) is one of important kinases in inflammation to phosphorylate inhibitor of nuclear factor kappa-B (IκBα) and then activate nuclear factor kappa-B (NF-κB). Inhibition of IKKβ has been a therapeutic strategy for inflammatory and autoimmune diseases. Here we report that IKKβ is constitutively activated in healthy donors and healthy Ikkβ C46A (cysteine 46 mutated to alanine) knock-in mice although they possess intensive IKKβ-IκBα-NF-κB signaling activation. These indicate that IKKβ activation probably plays homeostatic role instead of causing inflammation. Compared to Ikkβ WT littermates, lipopolysaccharides (LPS) could induce high mortality rate in Ikkβ C46A mice which is correlated to breaking the homeostasis by intensively activating p-IκBα-NF-κB signaling and inhibiting phosphorylation of 5' adenosine monophosphate-activated protein kinase (p-AMPK) expression. We then demonstrated that IKKβ kinase domain (KD) phosphorylates AMPKα1 via interacting with residues Thr183, Ser184, and Thr388, while IKKβ helix-loop-helix motifs is essential to phosphorylate IκBα according to the previous reports. Kinase assay further demonstrated that IKKβ simultaneously catalyzes phosphorylation of AMPK and IκBα to mediate homeostasis. Accordingly, activation of AMPK rather than inhibition of IKKβ could substantially rescue LPS-induced mortality in Ikkβ C46A mice by rebuilding the homeostasis. We conclude that IKKβ activates AMPK to restrict inflammation and IKKβ mediates homeostatic function in inflammation via competitively phosphorylating AMPK and IκBα.
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AIMS: We designed a case-control study to investigate the effect of vitamin D receptor gene (VDR) gene single nucleotide polymorphisms (SNPs) and possible gene- environment interaction on the susceptibility of renal cell carcinoma (RCC). METHODS: Generalized multifactor dimensionality reduction (GMDR) was used to find out the interaction combinations between SNPs and environmental factors, including gene- gene synergy and gene environment synergy effect. Logistic regression was used to analyze the correlation between the four SNPs in VDR gene and RCC, and the significant interaction combinations found by GMDR model were analyzed by hierarchical analysis. RESULTS: The genotype distribution of the control group was in accordance with Hardy- Weinberg equilibrium. Logistic regression analysis showed that the risk of RCC in VDR-rs7975232 A allele carriers was significantly higher than that of CC genotype carriers (CA + AA vs. CC), adjusted OR (95 % CI) = 1.75 (1.26-2.28). We used GMDR model to screen the best synergistic model between the four SNPs of VDR gene and smoking and drinking. We found a significant two locus model (P = 0.0010) involving rs7975232 and smoking. The cross- validation consistency of the two- locus model was 10/ 10, and the accuracy was 60.72 %. Compared with non-smokers with rs7975232 -CA or AA genotype, smokers with rs7975232 -CC genotype had the highest risk of RCC, or (95 % CI) = 2.23 (1.42-3.09), after adjustment for covariates. CONCLUSIONS: We found that the A allele of rs7975232 within VDR gene, interaction between rs7975232 and smoking were all associated with increased RCC risk.
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Objective:To investigate the feasibility of early off-bed mobility in patients with mechanical ventilation and its effect on delirium and the duration of delirium in the intensive care unit (ICU).Methods:Adult patients who were admitted to ICU of the Affiliated Hospital of Zunyi Medical University from January 1st to December 31st 2020 for invasive mechanical ventilation and no early activity contraindication were selected. The patients were randomly divided into two groups. The experimental group conducted early off-bed mobility, such as using the shift machine off-bed sitting and walking aids to assist standing and walking, and the off-bed mobility is based on patient tolerance. The control group was given early bed activities, including conducting the joint range activity, limb movement, bed sitting, upper limb elastic belt movement, and lower limb cycling, once a day. Each joint moved 15-20 times, a total of 30 minutes. Both groups were treated with anti-infection, mechanical ventilation, analgesia and sedation, and nutrition therapy. After intervention, confusion assessment method for the ICU (CAM-ICU) was used to assess the onset and duration of delirium, physical restraint rate and duration of physical restraint, mechanical ventilation time, and the length of ICU stay.Results:After excluding patients who died or gave up treatment during the intervention period, 266 patients were included, with 133 patients in the experimental group and 133 patients in the control group. There were no significant differences in gender, age, diagnosis, degree of illness, sedative drugs between the two groups. The incidence of the delirium in intervention group was significantly lower than that in control group [26.3% (35/133) vs. 42.1% (56/133), χ 2 = 7.366, P = 0.007], the duration of delirium was shorter than that in control group (hours: 11.26±4.11 vs. 17.00±3.29, t = -4.157, P = 0.000), the rate of physical restraint was lower than that in control group [19.5% (26/133) vs. 45.1% (60/133), χ 2 = 19.864, P = 0.000], the duration of physical restraint was shorter than that in control group (hours: 9.71±4.07 vs. 13.55±7.40, t = -5.234, P = 0.000), the mechanical ventilation time and the length of ICU stay were shorter than those in control group [mechanical ventilation time (hours) : 106.23±42.25 vs. 133.10±41.88, t = -3.363, P = 0.001; length of ICU stay (days) : 8.35±6.21 vs. 13.25±9.98, t =-4.190, P = 0.000]. Conclusions:Early off-bed mobility can reduce physical restraint rate and the incidence of delirium, and thus can accelerate rehabilitation in critically ill patients. Early off-bed mobility is safe and effective for patients with mechanical ventilation in ICU.
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The thioredoxin system is composed of thioredoxin (Trx), thioredoxin reductase (TR) and reduced nicotinamide adenine dinucleotide phosphate. Trx is an important antioxidant molecule that can resist cell death caused by various stresses and plays a prominent role in redox reactions. TR is a protein containing selenium (selenocysteine), mainly in three forms, i.e. TR1, TR2 and TR3. TR1 mainly distributed in the cytoplasm, TR2 mainly distributed in the mitochondria, and TR3 mainly distributed in the testes. TR can regulate cell growth and apoptosis. After the cell becomes cancerous, the expression of TR increases to promote cell growth and metastasis. Trx system is closely related to neurodegenerative diseases, parasitic infections, acquired immunodeficiency syndrome, rheumatoid arthritis, hypertension, myocarditis and so on. The Trx system can remove the reactive oxygen species (ROS) in the body, keep the inside and outside of the cell in a balanced state, and it interacts with the thioredoxin interacting protein (TXNIP), which plays an important role in the regulation of glucose metabolism and tumor treatment. The Trx system is an important target for drug treatment of many diseases. In this paper, the research progress of the thioredoxin system was reviewed.
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OBJECTIVE@#To analyze a pathogenic variant of MEFV gene in a family with autosomal dominant-familial Mediterranean fever (AD-FMF).@*METHODS@#A 5-year-old boy presented with recurrent aseptic meningitis and his major symptoms included recurrent fever with headache and vomiting. His family members including his mother, sister and brother also had recurrent fever. A genetic disease was considered. DNAs were extracted from patient and all his family members' blood samples. Whole exome sequencing was performed to identify putative pathogenic variants that can explain this family's condition and Sanger sequencing was conducted. The impact of detected variants were predicted and validated by bioinformatics.@*RESULTS@#A missense variant c.2229C>G (p.Phe743Leu) in MEFV gene was identified in the proband and his family members including his mother, sister and brother. This variant had not been reported in China previously, but the locus of it had already been reported in Arabic patient with AD-FMF (PS1). This variant was absent in major allele frequency databases (PM2) and had been predicted to be pathogenic based on Mutationtaster, PROVEAN and PolyPhen-2. In addition, the change of amino acid, locating in 743 locus of pyrin protein, encoding by MEFV gene, was found to cause SPRY_PRY_TRIM20 and SPRY_superfamily domain destroyed and finally influenced the fuction of pyrin protein. On the other hand, using UCSF chimera software, we find the variant c.2229C>G (p.Phe743Leu) can induce serious influence to the spatial structure of pyrin protein and loss of protein fuction (PP3). According to the ACMG variant classification guideline, the variant c.2229C>G (p.Phe743Leu) in MEFV gene was classified as likely pathogenic (PS1+PM2+PP3).@*CONCLUSION@#The condition of this AD-FMF family may be attributed to the missense variant c.2229C>G (p.Phe743Leu) in MEFV gene. The recurrent aseptic meningitis was a very rare manifestation in AD-FMF patients and had not been reported in China previously. The clinical and genetic findings of the present study are helpful for the further understanding of AD-FMF.
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Child, Preschool , Humans , Male , Familial Mediterranean Fever/genetics , Gene Frequency , Genetic Testing , Mutation , Pyrin/genetics , Exome SequencingABSTRACT
Objective:To summarize the perioperative nursing experience of a patient with hemophilia arthritis complicated with hepatitis C undergoing unilateral hip and knee arthroplasty at the same time.Methods:In August 2018, a patient with hemophilic arthritis complicated with hepatitis C was admitted to the first hospital of Lanzhou University. Preparations for health education, functional exercises, examination and evaluation before operation was made. Unilateral hip and knee arthroplasty were performed at the same time, strengthen the management of incisions and catheters after the operation, actively prevent complications, and guide patients to perform functional rehabilitation exercises under the conditions of good pain management and safe coagulation factor activity; pay attention to the nursing of coagulation factor replacement treatment in the whole perioperative period.Results:After careful nursing, no serious complications occurred during the patient's hospitalization, and the position and force line of the prosthesis were satisfactory by imaging examination before discharge.Conclusion:The perioperative care of this patient proved that the above-mentioned nursing measures are effective for hemophilic arthritis patients with hepatitis C undergoing unilateral hip and knee arthroplasty at the same time.
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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections have resulted in a number of severe cases of COVID-19 and deaths worldwide. However, knowledge of SARS-CoV-2 infection, diseases and therapy remains limited, underlining the urgency of fundamental studies and drug development. Studies have shown that induction of autophagy and hijacking of autophagic machinery are essential for infection and replication of SARS-CoV-2; however, the mechanism of this manipulation and function of autophagy during SARS-CoV-2 infection remain unclear. In the present study, we identified ORF3 as an inducer of autophagy and revealed that ORF3 localizes to the ER and induces FAM134B-related ERphagy through the HMGB1-Beclin1 pathway. As a consequence, ORF3 induces ER stress and inflammatory responses through ERphagy and sensitizes cells to ER stress-induced cell death, suggesting that SARS-CoV-2 ORF3 hijacks ERphagy and then harms ER homeostasis to induce inflammatory responses through excessive ER stress. These findings reveal a sequential induction of ERphagy, ER stress and acute inflammatory responses during SARS-CoV-2 infection and provide therapeutic potential for ERphagy and ER stress-related drugs for COVID-19 treatment and prevention. ImportanceSARS-CoV-2 infection and replication require autophagosome-like double-membrane vacuoles. Inhibition of autophagy suppresses viral replication, indicating the essential role of autophagy in SARS-CoV-2 infection. However, how SARS-CoV-2 hijacks autophagy and the function of autophagy in the disease progression remain unknown. Here, we reveal that SARS-CoV-2 ORF3 induces ERphagy and consequently induces ER stress to trigger acute inflammatory responses and enhance sensitivity to ER stress-induced apoptosis. Our studies uncover ERphagy-induced inflammatory responses during SARS-CoV-2 infection and provide a promising therapeutic approach for treating SARS-CoV-2 infection and inflammatory responses in COVID-19 by manipulating autophagy and ER stress.
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Melanoma differentiation-associated gene-5 (MDA5) acts as a cytoplasmic RNA sensor to detect viral dsRNA and mediates type I interferon (IFN) signaling and antiviral innate immune responses to infection by RNA viruses. Upon recognition of viral dsRNA, MDA5 is activated with K63-linked polyubiquitination and then triggers the recruitment of MAVS and activation of TBK1 and IKK, subsequently leading to IRF3 and NF-{kappa}B phosphorylation. Great numbers of symptomatic and severe infections of SARS-CoV-2 are spreading worldwide, and the poor efficacy of treatment with type I interferon and antiviral agents indicates that SARS-CoV-2 escapes from antiviral immune responses via an unknown mechanism. Here, we report that SARS-CoV-2 nonstructural protein 8 (NSP8) acts as an innate immune suppressor and inhibits type I IFN signaling to promote infection of RNA viruses. It downregulates the expression of type I IFNs, IFN-stimulated genes and proinflammatory cytokines by binding to MDA5 and impairing its K63-linked polyubiquitination. Our findings reveal that NSP8 mediates innate immune evasion during SARS-CoV-2 infection and may serve as a potential target for future therapeutics for SARS-CoV-2 infectious diseases. ImportanceThe large-scale spread of COVID-19 is causing mass casualties worldwide, and the failure of antiviral immune treatment suggests immune evasion. It has been reported that several nonstructural proteins of severe coronaviruses suppress antiviral immune responses; however, the immune suppression mechanism of SARS-CoV-2 remains unknown. Here, we revealed that NSP8 protein of SARS-CoV-2 directly blocks the activation of the cytosolic viral dsRNA sensor MDA5 and significantly downregulates antiviral immune responses. Our study contributes to our understanding of the direct immune evasion mechanism of SARS-CoV-2 by showing that NSP8 suppresses the most upstream sensor of innate immune responses involved in the recognition of viral dsRNA.
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Clinical symptoms of coronavirus disease 2019 (COVID-19) range from asymptomatic to severe pneumonia and death. Detection of individuals at high risk for critical condition is crucial for control of the disease. Herein, for the first time, we profiled and analyzed plasma cell-free DNA (cfDNA) of mild and severe COVID-19 patients. We found that in comparison between mild and severe COVID-19 patients, Interleukin-37 signaling was one of the most relevant pathways; top significantly altered genes included POTEH, FAM27C, SPATA48, which were mostly expressed in prostate and testis; adrenal glands, small intestines and liver were tissues presenting most differentially expressed genes. Our data thus revealed potential tissue involvement, provided insights into mechanism on COVID-19 progression, and highlighted utility of cfDNA as a noninvasive biomarker for disease severity inspections. One Sentence SummaryCfDNA analysis in COVID-19 patients reveals severity-related tissue damage.
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Objective:To assess the interobserver and inter-modalities agreement with two non-invasive diagnostic modalities of hepatocellular carcinoma in high-risk patients: contrast-enhanced ultrasound liver imaging reporting and data system (CEUS LI-RADS) and magnetic resonance imaging liver imaging reporting and data system (MRI LI-RADS).Methods:From August 2017 to August 2019, the CEUS and MRI data of patients at high risk for HCC from the Second Affiliated Hospital of Zhejiang University School of Medicine were analyzed retrospectively. A total of 217 lesions in 173 patients were classified according to CEUS LI-RADS v. 2017 or MRI LI-RADS v. 2018, by 4 blinded independent observers with more than 10 years of experience of CEUS or MRI. Interobserver and inter-modalities agreement was assessed with Cohen′s kappa.Results:The interobserver agreement was moderate and comparable for CEUS/MRI LI-RADS category (κ=0.606/0.603), the inter-modalities agreement was moderate for CEUS and MRI LI-RADS category (κ=0.564), LI-RADS 3, M, 4 and 5 by two imaging methods showed that the Kappa values were 0.739, 0.551, 0.734 and 0.592, respectively.Conclusions:The total inter-modalities agreement between CEUS and MRI LI-RADS categories is moderate, while the agreements of LI-RADS 3, 4 are strong, and LI-RADS M, 5 are moderate.
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In the practical teaching of large-scale medical equipment courses in medical colleges and universities. There are some problems such as less opportunities for students to operate the equipment which is possessed by teaching hospitals, the large-scale medical equipment in teaching hospitals are in a pathogenic environment, and some equipment such as CT, DR have radiation hazards, which lead to difficulties in practical teaching. These problems restrict the cultivation of students' practical ability, cause disconnections between theoretical teaching and practical teaching, and are not conducive to training "new engineering" and "new medical science" talents. So, this paper puts forward an idea which introduces virtual simulation teaching instruments of large-scale medical equipment into the teaching of medical equipment in medical colleges and universities, uses the physical structures, control systems and functional software of virtual simulation teaching instruments to implement the teaching of "integration of theory and practice", and also sets up validation experiments, comprehensive experiments, designing experiments and research explorative experiments in the teaching process. This kind of teaching can not only realize the high integration of theoretical teaching and practical teaching, but also consolidate students' basic theoretical knowledge, improve their abilities to solve complex engineering problems, and cultivate "new engineering" and "new medical" talents in line with the needs of the new era.
ABSTRACT
OBJECTIVE@#To investigate the effects of ALKBH5 on migration, invasion and epithelial-mesenchymal transition (EMT) of human trophoblast cells.@*METHODS@#The expression plasmid of ALKBH5 or a negative control plasmid (ALKBH5-NC) was transfected in human trophoblast HTR-8 /SVneo cells, and the expressions of ALKBH5 mRNA and protein were detected by qRT-PCR and Western blotting. Transwell assay was used to assess the changes in migration and invasion abilities of the trophoblast cells after the transfection. Western blotting was performed to detect the expressions of EMT-related proteins in the cells including vimentin, fibronectin, E-cadherin, N-cadherin, MMP9 and MMP2.@*RESULTS@#ALKBH5 mRNA and protein expressions were significantly higher in ALKBH5 group than in the control group (@*CONCLUSIONS@#ALKBH5 is involved in the pathogenesis of preeclampsia by inhibiting EMT of trophoblast cells and hence reducing their migration and invasion abilities.
