Stimulation of mouse hematopoietic stem cells by angiogenin and DNA preparations

Immature hematopoietic progenitors are a constant source for renewal of hemocyte populations and the basic component of the tissue and cell repair apparatus. A unique property of these cells of internalizing extracellular double-stranded DNA has been previously shown. The leukostimulatory effect demonstrated in our pioneering studies was considered to be due to the feature of this cell. In the present research, we have analyzed the effects of DNA genome reconstructor preparation (DNAgr), DNAmix, and human recombinant angiogenin on both hematopoietic stem cells and multipotent progenitors. Treatment with bone marrow cells of experimental mice with these preparations stimulates colony formation by hematopoietic stem cells and proliferation of multipotent descendants. The main lineage responsible for this is the granulocyte-macrophage hematopoietic lineage. Using fluorescent microscopy as well as FACS assay, co-localization of primitive c-Kit- and Sca-1-positive progenitors and the TAMRA-labeled double-stranded DNA has been shown. Human recombinant angiogenin was used as a reference agent. Cells with specific markers were quantified in intact bone marrow and colonies grown in the presence of inducers. Quantitative analysis revealed that a total of 14,000 fragment copies of 500 bp, which is 0.2% of the haploid genome, can be delivered into early progenitors. Extracellular double-stranded DNA fragments stimulated the colony formation in early hematopoietic progenitors from the bone marrow, which assumed their effect on cells in G0. The observed number of Sca1+/c-Kit+ cells in colonies testifies to the possibility of both symmetrical and asymmetrical division of the initial hematopoietic stem cell and its progeny.

Cell-to-Cell Communications of cGAS-STING Pathway in Tumor Immune Microenvironment / 浙江大学学报·医学版

Targeting cGAS-STING pathway is a promising strategy in tumor treatment. The pattern recognition receptor cGAS identifies dsDNA and catalyzes the formation of the second messenger 2'3'-cGAMP, activating the downstream interferons and pro-inflammatory cytokines through the adaptor protein STING. Notably, in tumor immune microenvironment, key components of cGAS-STING pathway are transferred among neighboring cells. The intercellular transmission under these contexts serves to sustain and amplify innate immune responses while facilitating the emergence of adaptive immunity. The membrane-based system, including extracellular vesicles transport, phagocytosis and membrane fusion transmit dsDNA, cGAMP and activated STING, enhancing the immune surveillance and inflammatory. The membrane proteins, including specific protein channel and intercellular gap junctions, transfer cGAMP and dsDNA, which are crucial to regulate immune responses. And the ligand-receptor interactions for interferons transmission amplifies the anti-tumor response. This review elaborates on the regulatory mechanisms of cell-to-cell communications of cGAS-STING pathway in tumor immune microenvironment. We further explore how these mechanisms modulate immunological processes and discuss potential interventions and immunotherapeutic strategies targeting these signaling cascades.

Expression of NF45 in laryngeal squamous cell carcinoma and its effect on radiation sensitivity of tumor cells / 中华放射肿瘤学杂志

Objective:To investigate the expression of double-stranded RNA-binding protein nuclear factor 45 (NF45) in laryngeal squamous cell carcinoma (LSCC), and the effect of NF45 on the radiation sensitivity of LSCC cells and its mechanism.Methods:NF45 expression in LSCC and adjacent tissues was detected by real-time reverse transcription PCR (qRT-PCR) and immunohistochemical staining. The NF45-ShRNA lentivirus was transfected into Hep-2 cells, and cell transfection efficiency was determined by qRT-PCR and Western blot . Hep-2 cells were randomly divided into the control group, 2 Gy group, sh-NC+2 Gy group and sh-NF45+2 Gy group. Lentivirus infection and 2Gy X-ray irradiation treatment were carried out. Cell proliferation activity was assessed by CCK-8 assay. Apoptosis rate was determined by flow cytometry. Hep-2 cells in each group were treated with mCherry-EGFP-LC3B. The levels of autophagy were detected by immunofluorescence staining. The ratio of autophagy-related protein microtubule-associated protein 1 light chain 3 (LC3)-Ⅱ/LC3-Ⅰ and the expression levels of Beclin-1 and p62 proteins were determined by Western blot.Results:The expression level of NF45 in LSCC tissues was significantly higher than that in adjacent tissues ( P<0.01). The relative expression levels of NF45 mRNA and protein in Hep-2 cells infected with NF45-shRNA were significantly lower than those in the control and sh-NC groups (all P<0.05). Compared with the control group, the cell proliferation activity was decreased, the apoptosis rate was increased, the intracellular autophagy-lysosome were increased, the ratio of LC3-Ⅱ/LC3-Ⅰ was increased, the relative expression levels of Beclin-1 protein were up-regulated, and the relative expression levels of p62 protein were down-regulated in the 2 Gy, sh-NC+2 Gy and sh-NF45+2 Gy groups (all P<0.05). Compared with the 2 Gy group, the cell proliferation activity was decreased, the apoptosis rate was increased, the intracellular autophagy lysosomes were increased, the LC3-Ⅱ/LC3-Ⅰ ratio was increased, the relative expression of Beclin-1 protein was up-regulated, and the relative expression of p62 protein was down-regulated in the sh-NF45+2 Gy group (all P<0.05). Conclusions:The expression of NF45 is up-regulated in LSCC tissues. Targeted down-regulation of NF45 expression can inhibit the proliferation activity of LSCC cells, promote cell apoptosis, and improve the sensitivity of tumor cells to radiation. The mechanism may be related to the regulation of autophagy levels.

Effects of GNG4 on DNA damage repair and chemosensitivity of ovarian cancer cisplatin-resistant A2780/DDP cells / 肿瘤研究与临床

Objective:To investigate the correlation of GNG4 with DNA damage repair and chemosensitivity of ovarian cancer cisplatin-resistant A2780/DDP cells.Methods:A2780/DDP cells were divided into 500 ng/ml cisplatin group (cDDP group), short hairpin RNA (shRNA)-GNG4 silencing GNG4 expression group (shRNA group), 500 ng/ml cisplatin and shRNA-GNG4 intervention group (shRNA+cDDP group), and non cisplatin and shRNA-GNG4 intervention group (blank control group). Western blot was used to detect the expressions of GNG4 and γH2AX proteins in each group; DNA damage in each group was detected by single cell gel electrophoresis. The focus formation of γH2AX gene at the injury site was detected by immunofluorescence. The ability of cell clone formation was detected by plate clone formation experiment.Results:Compared with the other three groups, the expression level of GNG4 protein in shRNA+cDDP group was the lowest, the expression level of γH2AX protein was the highest, and the differences were statistically significant (all P < 0.01). Single cell gel electrophoresis assay showed that the comet tail DNA% in blank control group, cDDP group, shRNA group and shRNA+cDDP group were (7.7±2.5)%, (12.3±3.6)%, (20.1±2.1)%, (38.6±2.8)%, respectively, and Olive trailing distance were 5.12±1.89, 8.23±2.97, 14.99±3.65, 22.43±3.17, respectively, the comet tail DNA% and Olive tail distance in shRNA+cDDP group were higher than those in the other three groups, and the differences were statistically significant (all P < 0.05). Immunofluorescence assay showed that the focus numbers of γH2AX in each cell of blank control group, cDDP group, shRNA group and shRNA+cDDP group were 4.2±0.7, 5.1±0.5, 26.8±3.3, 71.3±6.2, respectively, the shRNA+cDDP group was higher than the other three groups, and the differences were statistically significant (all P < 0.05). The clone formation rates of blank control group, cDDP group, shRNA group and shRNA+cDDP group were (78.27±5.01)%, (45.67±3.29)%, (26.20±5.76)%, (1.56±0.21)%, respectively, the shRNA+cDDP group was lower than the other three groups, and the differences were statistically significant (all P < 0.001). Conclusions:Down-regulation of GNG4 expression can increase the cisplatin sensitivity of ovarian cancer A2780/DDP cells, which may be achieved by inhibiting the DNA damage repair function induced by cisplatin.

DNA damage repair and cell cycle arrest / 国际生物医学工程杂志

In the face of DNA damage caused by various factors, cells have a set of response and repair mechanisms. Cell cycle arrest plays an important role in the DNA damage repair, which provides enough time for repairing damaged DNA. Research on cell cycle regulation focuses on cyclin-dependent protein kinases (CDKs) and cell cycle checkpoints. In the process of DNA damage repair, phosphatidylinositol-3-kinase like kinases (PIKKs) which are recruited to the DNA damage sites can activate cell cycle checkpoint-related proteins to halt cell cycle. In the common DNA damage repair pathways, such as base excision repair (BER), nucleotide excision repair (NER) , mismatch repair (MMR) , and DNA double-strand break repair, the recruitment of repair-related proteins also plays a role in the cell cycle regulation. In this paper, the relationship between the main forms of DNA damage repair and cell cycle arrest and relevant research progress were reviewed.

Post-transcriptional gene silencing: Basic concepts and applications

Post-transcriptional gene silencing (PTGS)-mediated gene silencing exploits the cellular mechanism whereintranscripts having sequence similarity to the double-stranded RNA (dsRNA) molecules present in the cell will besubjected to degradation. PTGS is closely related to natural processes such as RNA-mediated virus resistance andcross-protection in plants. Gene silencing and the cellular machinery for affecting this phenomenon might haveevolved as a natural protective measure against viral infection in plants. In PTGS, small interfering RNA (siRNA)molecules of 21–23 nucleotides length act as homology guides for triggering the systemic degradation of transcriptshomologous to the siRNA molecules. PTGS phenomenon, first discovered in transgenic petunia plants harbouringchalcone synthase gene and termed co-suppression, has been subsequently exploited to target specific gene transcripts for degradation leading to manifestation of desirable traits in crop plants. Targeted gene silencing has beenachieved either through the introduction of DNA constructs encoding dsRNA or antisense RNA or by deploying cosuppression constructs producing siRNAs against the transcript of interest. Understanding the mechanism of genesilencing has led to the development of several alternative strategies for inducing gene silencing in a precise andcontrolled way. This has paved the way for using PTGS as one ofthe chief functional genomicstools in plants and hashelped in unraveling the mechanism of many cellular processes and identifying the focal points in pathways, besides,opening new vistas in genetic engineering of plants for human benefits. PTGS has shown great potential in silencingthe deleterious genes efficiently so that value-added plant products could be obtained. Thus, PTGS has ushered in anew era in the genetic manipulation of plants for both applied and basic studies. In this review, we have outlined thebasics of RNAi-mediated gene silencing and summarized the work carried out at our institute using this approach, ascase studies. In particular, adopting RNAi-mediated gene silencing (a) as a method to restore fertility in transgenicmale sterile lines developed based on orfH522 gene from sunflower PET1-CMS source, (b) as a tool to suppress theproduction of toxic proteins, ricin and RCA, in castor, and (c) as an approach to induce bud necrosis virus resistancein sunflower has been discussed. Examples from other plant systems also have been mentioned to exemplify theconcept and utility of gene silencing in crop plants.

Alopecia areata y vitamina D sérica en pacientes iraquíes: un estudio de casos y controles / Alopecia Areata And Serum Vitamin D in Iraqi Patients: a case-control study

Background: Alopecia areata (AA) is a typical hair issue, which may have obliterating mental and social outcomes and is portrayed by the nearness of nonscarring alopecia. Objective: This examination has targets to assess the serum nutrient D levels , with AA; contrast the outcome and clearly sound control; and confirm relationship between AA types and serum nutrient D levels. Patients Also Methods: the examine might have been led clinched alongside Tikrit educating healing facility throughout those time starting with June 2019 of the limit for January 2020. Irrefutably the quantity of subjects associated with the assessment was ninety individuals isolated in two social events; the patients bundle were forty five the people who whimper of AA while the resulting gathering including a forty five age and sex-made solid volunteers were picked as a benchmark gathering. The degree and movement of the alopecia were noted and the patients were meticulously broke down for signs of various ailments. Research center assessments were led to patients and also to those control population, these included serum vitamin D levels were measured as 25-hydroxyvitamin D {25(OH)D} using a chemiluminescence microparticle immunoassay. Blood models were gotten starting with patients and control subjects after totally taught consent was gotten. Results : An essential complexity may have been found for serum 25-OH Vit D levels between patients other than controls. Vitamin D sufficiency were more common in controls than in patients. Serum Vitamin D was deficient in both cases and controls group; but, the deficiency was significantly more throughout AA group (35. 6%) compared to the handle group (11. 1%). Among the list patients gathering, levels associated with nutrient D were totally higher in guys in contrast with females. Conclusions: AA might be related with nutrient D deficiency as mean degrees of nutrient D of patients were seen as fundamentally lower than typical sound controls.

Electrochemical, spectroscopic, and molecular docking studies of the interaction between the anti-retroviral drug indinavir and dsDNA / 药物分析学报

In this study, an electrochemical DNA biosensor was developed using a straightforward methodology to investigate the interaction of indinavir with calf thymus double-stranded deoxyribonucleic acid (ct-dsDNA) for the first time. The decrease in the oxidation signals of deoxyguanosine (dGuo) and deoxy-adenosine (dAdo), measured by differential pulse voltammetry, upon incubation with different con-centrations of indinavir can be attributed to the binding mode of indinavir to ct-dsDNA. The currents of the dGuo and dAdo peaks decreased linearly with the concentration of indinavir in the range of 1.0-10.0μg/mL. The limit of detection and limit of quantification for indinavir were 0.29 and 0.98μg/mL, respectively, based on the dGuo signal, and 0.23 and 0.78μg/mL, respectively, based on the dAdo signal. To gain further insights into the interaction mechanism between indinavir and ct-dsDNA, spectroscopic measurements and molecular docking simulations were performed. The binding constant (Kb) between indinavir and ct-dsDNA was calculated to be 1.64 × 108 M-1, based on spectrofluorometric measure-ments. The obtained results can offer insights into the inhibitory activity of indinavir, which could help to broaden its applications. That is, indinavir can be used to inhibit other mechanisms and/or hallmarks of viral diseases.

CAS9 is a genome mutator by directly disrupting DNA-PK dependent DNA repair pathway

With its high efficiency for site-specific genome editing and easy manipulation, the clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR associated protein 9 (CAS9) system has become the most widely used gene editing technology in biomedical research. In addition, significant progress has been made for the clinical development of CRISPR/CAS9 based gene therapies of human diseases, several of which are entering clinical trials. Here we report that CAS9 protein can function as a genome mutator independent of any exogenous guide RNA (gRNA) in human cells, promoting genomic DNA double-stranded break (DSB) damage and genomic instability. CAS9 interacts with the KU86 subunit of the DNA-dependent protein kinase (DNA-PK) complex and disrupts the interaction between KU86 and its kinase subunit, leading to defective DNA-PK-dependent repair of DNA DSB damage via non-homologous end-joining (NHEJ) pathway. XCAS9 is a CAS9 variant with potentially higher fidelity and broader compatibility, and dCAS9 is a CAS9 variant without nuclease activity. We show that XCAS9 and dCAS9 also interact with KU86 and disrupt DNA DSB repair. Considering the critical roles of DNA-PK in maintaining genomic stability and the pleiotropic impact of DNA DSB damage responses on cellular proliferation and survival, our findings caution the interpretation of data involving CRISPR/CAS9-based gene editing and raise serious safety concerns of CRISPR/CAS9 system in clinical application.

CAS9 is a genome mutator by directly disrupting DNA-PK dependent DNA repair pathway

With its high efficiency for site-specific genome editing and easy manipulation, the clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR associated protein 9 (CAS9) system has become the most widely used gene editing technology in biomedical research. In addition, significant progress has been made for the clinical development of CRISPR/CAS9 based gene therapies of human diseases, several of which are entering clinical trials. Here we report that CAS9 protein can function as a genome mutator independent of any exogenous guide RNA (gRNA) in human cells, promoting genomic DNA double-stranded break (DSB) damage and genomic instability. CAS9 interacts with the KU86 subunit of the DNA-dependent protein kinase (DNA-PK) complex and disrupts the interaction between KU86 and its kinase subunit, leading to defective DNA-PK-dependent repair of DNA DSB damage via non-homologous end-joining (NHEJ) pathway. XCAS9 is a CAS9 variant with potentially higher fidelity and broader compatibility, and dCAS9 is a CAS9 variant without nuclease activity. We show that XCAS9 and dCAS9 also interact with KU86 and disrupt DNA DSB repair. Considering the critical roles of DNA-PK in maintaining genomic stability and the pleiotropic impact of DNA DSB damage responses on cellular proliferation and survival, our findings caution the interpretation of data involving CRISPR/CAS9-based gene editing and raise serious safety concerns of CRISPR/CAS9 system in clinical application.

Function and Characterization of the Argonaute2 Gene in the RNA Interference Pathway in the Diamondback Moth

Arch BiochemMol Biol2019;10(4):035-051DOI: 10.26502/abmb.007Archives of Biochemistry and Molecular BiologyVol. 10 No. 4 -December2019. 36AbstractArgonaute2 (AGO2) is a core catalytic component of the RNA-induces silencing complex (RISC) that binds to small guide RNAs containing small interfering RNA (siRNA) and microRNA (miRNA). The guide RNA leads RISC to the complementary mRNA for gene suppression. We cloned the full length cDNA (2193 bp) of the Ago2gene (PxAgo2) from diamondback moth (DBM, Plutella xylostella). The predicted PxAgo2 protein hadan83 kDa molecular weight withatheoretical isoelectric point of 9.39. The phylogenetic tree showed a high similarity of PxAgo2with Bombyx moriAgo2 (BmAgo2). Western blot and RT-qPCR analyses showed a clear increase in the PxAgo2 mRNA and protein expression levels in the egg, 4thinstar larva, pre-pupa, pupa and adult. The double-stranded RNA-mediated RNAi of PxAgo2in DBM larvae was found 3 h after dsRNA injection,and the knockdown level was increased over time up to 36 h. PxAgo2silencing recovered the expression of PxBurs-?(Bursicon-? inDBM) to the normal expression level, which was suppressed bydsPxBurs-?in a DBM cell line. The overexpression of PxAgo2fundamentally enhanced the PxBurs-? silencing efficiency in DBM cells. Our findings reveal that PxAgo2is involved in the dsRNA-regulated gene silencing mechanism and performs a crucial function in the RNAi process ofDBM

Expression of PD-1hiCXCR5-CD4+T cells in patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective To investigate the expression of peripheral programmed death (PD)-1hiCXCR5-CD4+T cells and its clinical significance in systemic lupus erythematosus (SLE). Methods Peripheral blood PD-1hiCXCR5-CD4+ T cells from 21 SLE patients and 16 healthy controls were examined by flow cytometry. The levels of serum anti-double-stranded deoxyribonucleic acid (dsDNA) antibodies were determined using immunoradiometric as-say. Data were analyzed with t test and Pearson's correlation test. Results The per-centages of PD-1hiCXCR5- cells within CD4+ T cell were significantly higher in SLE patients [(2.1 ±2.0)％] compared to normal controls [(0.3±0.3)％] (t=2.959, P<0.01). The percentages of PD-1hiCXCR5-cells within CD4+T cells in moderate to severe active SLE patients (3.0 ±2.0)％ was significantly increased compared to patients with mild or inactive (1.0±1.4)％(t=2.574, P<0.05) and normal controls (0.3±0.3)％ (t=5.149, P<0.01). The percentages of PD-1hiCXCR5- cells within CD4+ T cells from SLE patients were positively related with systemic lupus erythematosus disease activity index (SLEDAI) (r=0.475, P=0.0297). SLE patients in serum anti-dsDNA antibodies positive group (2.7±2.1)％displayed a higher percentage of PD-1hiCXCR5-cells within CD4+T cells than patients in serum anti-dsDNA antibodies negative group (0.6 ±0.5)％ (t=2.303, P<0.05). The percentages of PD-1hiCXCR5-cells within CD4+T cells from SLE patients were positively correlated with anti-dsDNA antibody titers. Conclusion The percentages of PD-1hiCXCR5- cells within CD4+ T cells from SLE patients are increased and are positively correlated with SLEDAI and anti-dsDNA antibody levels. Increased percentage of PD-1hiCXCR5-cells within CD4+T cells might play an important role in the pathogenesis of SLE.

p53 and its isoforms in DNA double-stranded break repair / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

DNA double-stranded break (DSB) is one of the most catastrophic damages of genotoxic insult. Inappropriate repair of DNA DSBs results in the loss of genetic information, mutation, and the generation of harmful genomic rearrangements, which predisposes an organism to immunodeficiency, neurological damage, and cancer. The tumor repressor p53 plays a key role in DNA damage response, and has been found to be mutated in 50% of human cancer. p53, p63, and p73 are three members of the p53 gene family. Recent discoveries have shown that human p53 gene encodes at least 12 isoforms. Different p53 members and isoforms play various roles in orchestrating DNA damage response to maintain genomic integrity. This review briefly explores the functions of p53 and its isoforms in DNA DSB repair.

p53 and its isoforms in DNA double-stranded break repair / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

DNA double-stranded break (DSB) is one of the most catastrophic damages of genotoxic insult. Inappropriate repair of DNA DSBs results in the loss of genetic information, mutation, and the generation of harmful genomic rearrangements, which predisposes an organism to immunodeficiency, neurological damage, and cancer. The tumor repressor p53 plays a key role in DNA damage response, and has been found to be mutated in 50% of human cancer. p53, p63, and p73 are three members of the p53 gene family. Recent discoveries have shown that human p53 gene encodes at least 12 isoforms. Different p53 members and isoforms play various roles in orchestrating DNA damage response to maintain genomic integrity. This review briefly explores the functions of p53 and its isoforms in DNA DSB repair.

Application value of anti-double stranded DNA antibody in systemic lupus erythematosus by enzyme-linked immunosorbent assay / 上海交通大学学报（医学版）

Objective • To investigate the characteristics and clinical application value of anti-double stranded DNA (dsDNA) antibody detected by enzyme-linked immunosorbent assay (ELISA). Methods • 186 patients with systemic lupus erythematosus, 183 autoimmune disease of non-SLE controls, 78 non-autoimmune disease controls and 50 healthy controls were selected. The serum anti-dsDNA antibody was detected simultaneously by the methods of ELISA and radioimmunoassay (RIA) and their diagnostic efficacies for detection were compared. Results • The sensitivities of anti-dsDNA antibody in SLE by RIA and ELISA were 47.31% and 62.90%, respectively. The specificities were 85.85% and 81.67%, respectively. The positive predictive were 66.67% and 67.24%, respectively. The negative predictive were 73.15% and 78.14%, respectively. The anti-dsDNA antibody levels of SLE patients detected by ELISA and RIA both increased with the increase of SLE disease activity index. Conclusion • The specificity of ELISA is similar with RIA in diagnosing SLE, and the sensitivity is higher than RIA, which can screen the patients with SLE. In addition, the two methods are both suitable for monitoring the condition of SLE.

An overlap case of Parry朢omberg syndrome and en coup de sabre with striking ocular involvement and anti-double-stranded DNA positivity

Parry朢omberg syndrome (PRS) may overlap localized scleroderma (morphea) lesions with linear depression (en coup de sabre [ECDS]). Overlap case with PRS and ECDS was presented. Enophthalmos, uveitis, ocular torticollis, keratic linear precipitates, and anti-double-stranded DNA positivity were identified. Subendothelial keratic precipitates detected by an in vivo laser scanning confocal microscopy were the first profiled in the literature. Patients must be evaluated and followed up carefully by their clinics to prevent misdiagnosis and unnecessary procedures such as surgery of ocular torticollis as muscular torticollis.

Lupus eritematoso sistémico pediátrico: reporte de un caso clínico / Systemic lupus erythematosus pediatric: report of a clinical case

El presente reporte de caso se hace con la finalidad de mejorar la valoración del paciente con esta enfermedad, así de esta manera realizar un diagnóstico precoz, porque que es una enfermedad autoinmune muy poco frecuente en pacientes pediátricos. Además, debido a que no se entienden por completo las causas del lupus eritematoso sistémico, no se sabe cómo prevenirlo pero los brotes de la enfermedad se pueden reducir. Actualmente existe una guía de práctica clínica para el tratamiento a partir de las conclusiones acordadas en la Federación Española de lupus eritematoso sistémico en 2017, pero la valoración del impacto en los resultados de los tratamientos depende de su cumplimiento, los cuales aún se encuentran en estudios

The present case report is made with the purpose of improving the assessment of the patient with this pathology, thus making an early diagnosis, because it is a very rare autoimmune disease in pediatric patients. In addition, because the causes of systemic lupus erythematosus are not fully understood, it is not known how to prevent it, but outbreaks of the disease can be reduced. Currently there is a clinical practice guide for treatment based on the conclusions agreed in the Spanish Federation of Systemic Lupus Erythematosus in 2017, but the assessment of the impact on the results of treatments depends on their compliance, which are still found in studies

Mechanism of migration and release of high mobility group box-1 / 中南大学学报(医学版)

High mobility group box-1 (HMGB1) is an evolutionarily conserved protein,which widely exists in mammals.HMGB 1 contains the nucleus localization sequences.Intracellular and extracellular HMGB1 shows different biological functions.Extracellular HMGB1 is closely related to sepsis,cancer,rheumatoid immune,atherosclerosis,ischemia-reperfusion injury and so on.The mobilization of HMGB 1 from the nucleus to the cytoplasm and subsequent release involves the processes of post-translation modification,active secretion and nuclear localization.

Effects of silencing single-stranded DNA-binding protein 1 gene on proliferation and DNA repair of submandibular gland cells after irradiation / 中华放射医学与防护杂志

Objective To investigate the effects of silencing the gene of single-stranded DNA-binding protein 1 (SSB1) on proliferation and DNA repair of rat submandibular gland (SMG) cells after irradiation, and explore the relationship between SSB1 and DNA damage repair. Methods Primary rat SMG cells were obtained by mechanical-enzyme digestion and identified by immunohistochemistry. The cells were divided into three groups, including blank control, negative control and shRNA transfection group. The shRNA was transfected into cells by recombinant adenovirus vector. Real-time quantitative PCR ( qRT-PCR) was used to detect the expression of SSB1 after silencing. The cell viability was detected by CCK-8 assay. Immunofluorescence analysis was performed to observe the dynamic formation of γ-H2AX foci. Results The SMG cells were positively stained for both Pan CK and α-Amylase. The efficiency of shRNA transfection was about 90%at 72 h post-transfection. Compared with the blank control group, the expression of SSB1 was significantly decreased in the cells transfected with shRNA (t=16. 24, P<0. 05). The cell viability of shRNA transfection group without irradiation was decreased indistinctively and became lower than the blank control group significantly until 120 h(t=3. 29, P<0. 05). After radiation with 5 Gy of γ-rays, the cell viability of shRNA transfection group was lower than that of the control groups significantly (F=10. 19-30. 13, P<0. 05). Silencing the expression of SSB1 could increase the number ofγ-H2AX foci in SMG cells at different time of radiation. Conclusions After silencing of the expression of SSB1, the SMG cells could be more radiosensitive, which indicats that SSB1 may play an important role in DNA damage repair after radiation.

Effects of silencing single-stranded DNA-binding protein 1 gene on proliferation and DNA repair of submandibular gland cells after irradiation / 中华放射医学与防护杂志

Objective To investigate the effects of silencing the gene of single-stranded DNA-binding protein 1 (SSB1) on proliferation and DNA repair of rat submandibular gland (SMG) cells after irradiation, and explore the relationship between SSB1 and DNA damage repair. Methods Primary rat SMG cells were obtained by mechanical-enzyme digestion and identified by immunohistochemistry. The cells were divided into three groups, including blank control, negative control and shRNA transfection group. The shRNA was transfected into cells by recombinant adenovirus vector. Real-time quantitative PCR ( qRT-PCR) was used to detect the expression of SSB1 after silencing. The cell viability was detected by CCK-8 assay. Immunofluorescence analysis was performed to observe the dynamic formation of γ-H2AX foci. Results The SMG cells were positively stained for both Pan CK and α-Amylase. The efficiency of shRNA transfection was about 90%at 72 h post-transfection. Compared with the blank control group, the expression of SSB1 was significantly decreased in the cells transfected with shRNA (t=16. 24, P<0. 05). The cell viability of shRNA transfection group without irradiation was decreased indistinctively and became lower than the blank control group significantly until 120 h(t=3. 29, P<0. 05). After radiation with 5 Gy of γ-rays, the cell viability of shRNA transfection group was lower than that of the control groups significantly (F=10. 19-30. 13, P<0. 05). Silencing the expression of SSB1 could increase the number ofγ-H2AX foci in SMG cells at different time of radiation. Conclusions After silencing of the expression of SSB1, the SMG cells could be more radiosensitive, which indicats that SSB1 may play an important role in DNA damage repair after radiation.