ABSTRACT
Objective To explore the protective effect of polypeptide Apelin on podocyte mitochondria in diabetic nephropathy and underling mechanisms.Methods Human renal podocytes were divided into four experimental groups:control group,high glucose(HG)group(glucose 25 mmol/L,48 h),Apelin intervention HG group(Ape-lin-13 1 μmol/L,48 h)and Apelin group(Apelin-13 1 μmol/L,48 h).The podocyte apoptosis was observed by TUNEL staining,the expression of mitochondrial membrane protein FUNDC1 was detected by Western blot,and the binding of mitochondrial fission protein DRP1 to FUNDC1 was examined by immunoprecipitation.The 8-week-old male mice were divided into three experimental groups:control group,diabetes group(intraperito-neal injection of streptozotocin 150 mg/kg,only one time)and Apelin intervention DM group(intraperitoneal injection of Apelin-13 0.3 μmol/kg,daily).The morphology of renal was observed by PAS staining and trans-mission electron microscopy.Plasma creatinine(Cr),urea nitrogen,urinary albumin and creatinine were de-tected by ELISA kit.The level of creatinine clearance rate(Ccr)and urinary albumin/creatinine(ACR)was calculated.Results Compared with the control group,the podocyte apoptosis and expression of FUNDC1 in the HG group increased significantly(P<0.05),and the combination of mitochondrial fission division protein DRP1 to FUNDC1 raised.Meanwhile,compared with the HG group,the number of apoptosis,the expression of FUNDC1(P<0.05),and the combination of DRP1 to FUNDC1 all reduced in Apelin intervention HG group.Animal experiments showed that the kidney structure of the control group was intact.In the DM group,the num-ber of podocytes decreased significantly,the foot processes were fused and dropped off.In the Apelin intervention DM group,podocyte lesions were less severe than those in the DM group.Compared with the control group,the level of plasma Cr,BUN and urine ACR in the DM group increased,while the level of Ccr decreased significantly(P<0.05).However,compared with the DM group,the level of above biomarkers in the Apelin intervention DM group was improved(P<0.05).Conclusions Apelin keeps mitochondrial homeostasis and reduces podocyte ap-optosis by inhibiting the expression of mitochondrial membrane protein FUNDC1,which may contribute to allevia-tion of diabetic nephropathy.
ABSTRACT
Objective:To analyze the epidemiological characteristics, diagnosis and treatment of malaria in the peacekeeping mission area of the Democratic Republic of the Congo (DRC), and to evaluate the efficacy of artemisinin based combination therapy (ACT), to provide clinical basis for the diagnosis and treatment of malaria.Methods:The clinical data of malaria-infected peacekeepers admitted to the Chinese Secondary Hospital of the United Nations Stabilization Mission in DRC (MONUSCO) from January 2014 to September 2020 were collected, and the general information, incidence characteristics, treatment and clinical outcomes of the patients were retrospectively analyzed.Results:From January 2014 to September 2020, 362 peacekeepers were hospitalized with malaria, the average annual incidence case was 54 cases per year, and the annual incidence was 9.5/1 000, with a median of 2.5 days (1 - 9 days) from onset to diagnosis. Severe malaria accounted for 7.73% (28/362) and uncomplicated malaria accounted for 92.27% (334/362). The incidence rate was 37.57% (136/362) in the dry season (April to September) and 62.43% (226/362) in the rainy season (October to March of the following year). After ACT antimalarial treatment, all patients were cured clinically. Eight cases recurred and were cured clinically after drug conversion ACT retreatment.Conclusions:In the peacekeeping mission area of DRC, peacekeepers are generally susceptible to malaria. ACT has a high cure rate, safety and efficiency in clinical treatment.
ABSTRACT
Gene editing is an advanced technique based on artificial nucleases and can precisely modify genome sequences. It has shown great application prospects in the field of medicine and has provided a new precision therapy for diseases. Primary immunodeficiency disease is a group of diseases caused by single gene mutation and characterized by recurrent and refractory infections, with an extremely high mortality rate. The application of gene editing has brought hope for curing these diseases. This article reviews the development of gene editing technology and briefly introduces the research and application of gene editing technology in primary immunodeficiency disease.
Subject(s)
Humans , Gene Editing , Primary Immunodeficiency DiseasesABSTRACT
AIM:To investigate the effects of aspirin on the apoptosis of nasopharyngeal carcinoma cell lines CNE2R and CNE2 with different radioresistance and its potential mechanism. METHODS:The effects of aspirin on the cell viability, apoptosis, and protein levels of procaspase-3, cleaved caspase-3, procaspase-9, procaspase-12, PARP and cleaved PARP,PI3K p110α,Akt,Bcl-2,Bax and p27 in the CNE2R cells and CNE2 cells were detected by the methods of MTT assay,flow cytometry and Western blot. RESULTS:Aspirin inhibited the viability of homologous nasopharyngeal carcinoma cell lines CNE2 and CNE2R(with the IC50to CNE2 cells of 6.18,3.92 and 3.06 mmol/L for 24 h,48 h and 72 h,respectively;and with the IC50to CNE2R cells of 7.05,3.90 and 2.20 mmol /L for 24 h,48 h and 72 h,respec-tively). After treated with aspirin for 48 h, the apoptotic rate of CNE2R cells was higher than that of CNE2 cells (P<0.05). After treated with aspirin for 48 h,the protein levels of procaspase-3,procaspase-9,procaspase-12 and PARP were decreased,the protein levels of cleaved caspase-3,cleaved PARP and p27 were increased,and the protein levels of PI3K p110α,Akt and Bcl-2/Bax were decreased. CONCLUSION:Aspirin inhibits the viability of homologous nasopharyngeal carcinoma cell lines CNE2R and CNE2 with different radioresistance. Aspirin also induces the apoptosis of CNE2 and CNE2R cells,which is more effective in CNE2R cells. The underlying mechanisms may be involved in affecting PI3K/Akt signaling pathway,Bcl-2/Bax and p27 expression.
ABSTRACT
<p><b>OBJECTIVE</b>To observe the prevention and clinical efficacy of combination of Liuwei Dihuang Pill (LDP) and Ginkgo Leaf Tablet (GLT) for early diabetic retinopathy (DR).</p><p><b>METHODS</b>Using randomized, double-blind, double simulation, parallel controlled clinical trial, 140 type 2 diabetes mellitus (T2DM) outpatients were recruited and assigned to the treatment group and the control group, 70 in each group. All patients received basic Western medicine treatment (such as blood glucose and pressure control). Patients in the treatment group took LDP (8 pills each time, 3 times per day) and GLT (19.2 mg each time, 3 times per day), while those in the control group took LDP placebos and GLT placebos. All treatment lasted for 24 consecutive months. All subjects were followed-up every month. The general clinical data as sex, age, and metabolic data such as blood glucose, blood pressure, blood lipid, and DR prevalence rate were collected and statistically analyzed.</p><p><b>RESULTS</b>There was no significant difference in levels of blood glucose, blood pressure, or blood lipid between the two groups (P > 0.05). After treatment the DR incidence rate was significantly lower in the treatment group than in the control group [3.1% (2/64) vs 18.6% (11/59), P < 0.05)]. Meanwhile, the DR prevalence rate of the treatment group was also significantly lower than that of the control group [6.3% (4/64) vs 20.0% (13/59), P < 0.05].</p><p><b>CONCLUSION</b>Combination of LDP and GLT could effectively prevent and treat the development of DR in T2DM patients.</p>
Subject(s)
Humans , Blood Glucose , Blood Pressure , Diabetes Mellitus, Type 2 , Diabetic Retinopathy , Drug Therapy , Double-Blind Method , Drugs, Chinese Herbal , Therapeutic Uses , Ginkgo biloba , Chemistry , Phytotherapy , TabletsABSTRACT
MonoMAC syndrome is a newly discovered immune deficiency syndrome caused by GATA-2 mutation, which is an autosomal dominant genetic disease. MonoMAC syndrome has typical immune cell abnormalities, with severe infection and is prone to develop into a hematological disease. Therapeutics for this disease mainly relies on symptomatic treatment and hematopoietic stem cell transplantation. In this paper, the research advances in clinical manifestations, laboratory tests, pathogenesis, diagnosis and treatment of MonoMAC syndrome are reviewed.
Subject(s)
Humans , GATA2 Transcription Factor , Genetics , Immunologic Deficiency Syndromes , Genetics , Monocytes , Pathology , Mutation , Mycobacterium Infections , SyndromeABSTRACT
<p><b>OBJECTIVE</b>To detect the disparity of three cytokines interleukin-6 (IL-6), interferon-inducible protein 10 (IP-10) and interleukin-17 (IL-17) in peripheral blood (PB) and synovial fluid (SF) of patients with juvenile idiopathic arthritis (JIA).</p><p><b>METHOD</b>Serum concentrations of the three cytokines were measured in 27 patients with 13 systemic-onset JIA (sJIA), 14 polyarticular JIA (pJIA) and 28 healthy controls using enzyme-linked immunosorbent assay (ELISA). Nineteen patients with no marked arthritis symptom or only temporary arthralgia were enrolled in probable sJIA group. SF from 18 patients with 7 sJIA, 11 pJIA were examined for cytokine levels.</p><p><b>RESULT</b>(1) The statistically significant difference in serum IL-6 was detected between sJIA and healthy control group [28.0(4.2-59.2) ng/L vs. 12.3 (2.1-13.8) ng/L, P < 0.05], but no significant difference between probable sJIA and healthy control group [11.8(7.7-39.2) ng/L vs. 12.3 (2.1-13.8) ng/L, P > 0.05] was found. There were statistically significant differences between sJIA group and healthy control group in serum concentrations of IL-17 [14.0(9.8-34.3) ng/L vs. 9.8 (7.9-16.2) ng/L, P < 0.05], yet compared to healthy control group, no significant difference in concentration level of IL-17 was found in pJIA Group [14.2(9.9-16.9) ng/L vs. 9.8(7.9-16.2) ng/L, P > 0.05].(2) In sJIA and pJIA SF, the median IP-10 level was significantly higher compared to respective PB levels [619.7 (160.9, 873.1) ng/L vs. 64.8 (27.4-111.9) ng/L;660.9 (401.9, 1349.8) ng/L vs. 97.4 (41.9-222.1) ng/L, P < 0.01, respectively], but there was only significant difference in IL-17 between pJIA SF and PB [22.9 (17.1, 45.8) ng/L vs. 14.2 (9.9-16.9) ng/L, P < 0.01].</p><p><b>CONCLUSION</b>IL-6 may play more important role in the pathogenesis of sJIA. Moreover, IL-6 may be the biomarker associated with arthritis in early JIA stage. Both autoinflammation and autoimmune response may be involved in the pathogenesis of sJIA. IL-17 enrichment may only occur in local joint, the levels of IL-17 in PB may not be significantly increased. The prominent expression gradient between SF and PB of IP-10 maybe the basis of performing chemotaxis and further causing joint damage.</p>
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Male , Arthritis, Juvenile , Blood , Allergy and Immunology , Metabolism , Case-Control Studies , Chemokine CXCL10 , Blood , Metabolism , Enzyme-Linked Immunosorbent Assay , Interleukin-17 , Blood , Metabolism , Interleukin-6 , Blood , Metabolism , Knee Joint , Metabolism , Synovial Fluid , Allergy and Immunology , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To establish a novel flow cytometry-based assay for measuring the expression of lysosomal-associated membrane protein 1 (LAMP-1, CD107α) on the cell surface of natural killer (NK) cells and cytotoxic T lymphocyte (CTL) and evaluate the screening value of this assay for cytotoxic defects-related diseases such as familial hemophagocytic lymphopro-liferative (FHL) syndrome.</p><p><b>METHOD</b>Three suspected Chediak-Higashi Syndrome (CHS) patients, three suspected FHL patients and 10 healthy children were enrolled in the study from October 2010 to June 2011. Their PBMCs were separated and activated overnight with IL-2. After the granule release of NK cells activated by phytohemagglutinin (PHA) and CD8+T cells by anti-CD3, the CD107α expression were analyzed by flow cytometry. The peripheral blood DNA and RNA of the patients were extracted to analyze the pathogenic genes via DNA-PCR/RT-PCR and direct sequencing.</p><p><b>RESULT</b>The CD107α expression on CTL in the ten healthy children significantly increased after activation by anti-CD3 [(0.18 ± 0.07)% vs. (4.47 ± 2.36)%, P < 0.05] and NK cells after activation by PHA [(0.27 ± 0.07)% vs. (5.80 ± 2.83)%, P < 0.05]. The frequency of CD107α-expression NK cells in three suspected CHS after activation was significantly elevated when compared with the healthy control [0.5%, 0.6% vs. (5.80 ± 2.83)%] except patient 2. After the anti-CD3 activation, the frequency of CD107α expression on CTL cells also showed no significant difference [0.3%, 0.9%, 0.2% vs. (4.47 ± 2.36)%] in three patients. All of their mean fluorescence intensity (MFI) showed the same trend. Patient 1 and 3 were identified to have LYST mutations (Patient 1: c.5411-5414 del TTTC, L1741fsX1758 and c.7975 C > T, R2596X; Patient 3: c.4863G > A, R1563H and c.5392-5393delAA, E1739fsX1756). There was no mutation identified in the LYST gene for patient 2. CD107α expression of NK cells and CTL in the suspected FHL patients and in mirror of these findings, no underlying gene variation of PRF, MUNC13-4 and STX11 were identified.</p><p><b>CONCLUSION</b>We developed a method to quantitatively assess cytotoxicity of the NK cells and CTL by measuring the expression of CD107α on the cell membrane, which appeared to be an effective and rapid screening test for cytotoxic defects-related diseases such as FHL and other HLH secondary to primary immunodeficiency.</p>
Subject(s)
Child, Preschool , Female , Humans , Infant , Male , Case-Control Studies , Cell Degranulation , Allergy and Immunology , Cell Membrane , Metabolism , Chediak-Higashi Syndrome , Diagnosis , Genetics , Allergy and Immunology , Metabolism , Cytotoxicity, Immunologic , Flow Cytometry , Methods , Interleukin-2 , Metabolism , Killer Cells, Natural , Allergy and Immunology , Metabolism , Lymphohistiocytosis, Hemophagocytic , Diagnosis , Genetics , Allergy and Immunology , Metabolism , Lysosomal-Associated Membrane Protein 1 , Metabolism , Mutation , Phytohemagglutinins , Metabolism , T-Lymphocytes, Cytotoxic , Allergy and Immunology , MetabolismABSTRACT
<p><b>OBJECTIVE</b>X-linked lymphoproliferative disease (XLP), a genetic disorder characterized by immunodeficiency to Epstein-Barr virus (EBV) infection, has been linked to mutations in the SH2D1A gene. XLP patient displays EBV associated fulminant infectious mononucleosis or hemophagocytic lymphohistocytosis, hypogammaglobulinemia or malignant lymphoma. Here we report the clinical features, gene mutation and SAP expression on PBMCs of a Chinese patient with XLP and potential carriers.</p><p><b>METHOD</b>A 6 years old male patient and his maternal relatives were enrolled in this study. The patient was found to have with a renal Burkitt lymphoma on the right waist at 5 years of age by accident. His elder brother and a maternally related cousin both died of multiple systemic organ dysfunction syndrome (MODS) due to fulminant infectious mononucleosis (FIM) at the age of one year. The patient and his maternal relatives were subjected to detection of SAP expression on the PBMCs by flow cytometry and gene mutation analysis of SH2D1A by using PCR based on genomic DNA.</p><p><b>RESULT</b>The patient exhibited 536.9 copy/ml level of circulating EBV-DNA during remission. Sequence analysis showed that the patient harbored a nonsense mutation in exon 2 (C462T), resulting in a premature stop codon (Arg55X). His mother and some of the maternal relatives were proved to be carriers of this mutation. SAP expression from the patient was significantly reduced as compared to normal individual and the carriers.</p><p><b>CONCLUSION</b>We identified a Chinese XLP case genetically. Assessment of SAP expression on PBMCs by flow cytometry seemed to be an effective rapid diagnostic method for this disease. Absence of EBV infection does not diminish the possibility of XLP.</p>
Subject(s)
Child , Humans , Male , Carrier Proteins , Genetics , DNA, Viral , Blood , Epstein-Barr Virus Infections , Exons , Herpesvirus 4, Human , Intracellular Signaling Peptides and Proteins , Genetics , Lymphoproliferative Disorders , Genetics , Virology , Mutation , Pedigree , Signaling Lymphocytic Activation Molecule Associated ProteinABSTRACT
<p><b>OBJECTIVE</b>To investigate variation of FOXP3 and it's expression in male children presented with severe and early-onset enteropathy, rash with or without insulin-dependent diabetes mellitus (IDDM).</p><p><b>METHODS</b>Four male children presented with severe and early-onset enteropathy, rash, with or without IDDM were subjected to detection of FOXP3 expression on the PBMC by flow cytometry and FOXP3 gene analysis. The maternal gene analysis was subsequently performed once the variant FOXP3 gene was found. All 11 exons and splice sites within FOXP3 gene were amplified by polymerase chain reaction (PCR) from genomic DNA. Reverse transcription polymerase chain reaction was used to amplify the FOXP3 transcripts. Sequence analysis was performed directly on the bulk PCR products forwardly and reversely. The candidate mutation site was compared with that of 100 healthy controls to exclude polymorphism. Flow cytometry was used to determine FOXP3 expression on CD4+CD25+ T cells and the frequency of Tregs in CD4+ T cells.</p><p><b>RESULTS</b>One of the 4 patients showed a G13128A genetic variation in exon 11, which resulted in a Met370Ile substitution. No sequence variations were found at the same site in any of 100 healthy controls, indicating that the Met370Ile substitution is not a polymorphism but a novel missense mutation. The patient's mother was identified as a carrier for this mutation. There was no reduced frequency of Tregs in the peripheral blood of the patient and FOXP3 protein expression is normal as compared with controls.</p><p><b>CONCLUSION</b>A novel missense mutation of FOXP3 which causes IPEX phenotype was identified in a Chinese child according to immunologic screening and gene sequencing. Infants with early-onset IDDM and persistent diarrhea should be suspected as IPEX, FOXP3 gene analysis will be a reliable diagnostic approach to IPEX.</p>
Subject(s)
Child, Preschool , Humans , Infant , Male , DNA Mutational Analysis , Diabetes Mellitus, Type 1 , Genetics , Allergy and Immunology , Forkhead Transcription Factors , Genetics , Genes, X-Linked , Genetic Diseases, X-Linked , Genetics , Allergy and Immunology , Metabolism , Intestinal Diseases , Genetics , Metabolism , Mutation, Missense , Syndrome , T-Lymphocytes, Regulatory , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>Mutation in the interleukin-7 receptor-alpha (IL-7R alpha) chain causes a rare type of severe combined immunodeficiency (SCID) with presence of NK cells in the peripheral blood. Here we report the molecular and clinical characterization of a compound heterozygosity mutation in the interleukin-7 receptor-alpha gene that resulted in SCID in a patient firstly from China.</p><p><b>METHOD</b>A 5 month-old male patient and his parents were enrolled in this study. Since 15 days of age, the patient had had recurrent fever, persistent cough and diarrhea. He was in poor general condition with pyorrhea and ulceration of the BCG scar. His brother died of severe infection at 4 months of age. He was initially diagnosed as SCID according to clinical manifestation and immunological analysis. A panel of SCID candidate genes including IL-2RG, RAG1/RAG2 and IL-7R alpha of patient and his parents were amplified by polymerase chain reaction (PCR) from genomic DNA. Reverse transcription polymerase chain reaction (RT-PCR) was used to amplify the IL-7R alpha transcripts. Sequencing was performed directly on the PCR products forward and reversely.</p><p><b>RESULT</b>The serum immunoglobulin (Ig) profile was IgG 6867 mg/L (normal range, 3050 - 8870 mg/L); IgM 206 mg/L and IgA 249 mg/L, IgE 2.3 IU/ml (normal range < 150 IU/ml). The patient was treated with IVIG previously. There were no T-cells but increased percentage of B-cells (58%) and NK cells (42%) in the peripheral blood was found. Needle biopsies from enlarged axillary lymph node was identified positive for Mycobacterium bovis under microscope and by culture. The patient had a compound heterozygosity mutation in the IL-7R alpha gene:on one allele, there was a splice-junction mutation in intron 4 (intron 4(+1)G > A), for which his father was a carrier; whereas on the other allele, a nonsense mutation at position 638 in exon 5 with a premature stop codon (638 C > T, R206X) was identified, for which his mother was a carrier. The splice-junction mutation in intron 4 of IL-7R alpha was firstly reported. The IL-7R alpha mRNA expression of the patient was remarkably reduced whereas the parents had relatively normal IL-7R alpha mRNA expression. IL-7R alpha cDNA of the patient was amplified by nested PCR. The PCR products were purified, cloned with a TA Cloning Kit and sequenced directly. A 64 bp deletion was found in exon 4 of IL-7R alpha. No mutation was found in IL-2RG and RAG1/RAG2 of the patient and his parents.</p><p><b>CONCLUSION</b>This is the first case with a compound heterozygosity mutation in the IL-7R receptor alpha gene and T-B+NK+ phenotype from China. Intron 4(+1)G > A was a novel mutation.</p>
Subject(s)
Humans , Infant , Male , DNA , Genome, Human , Heterozygote , Mutation , Receptors, Interleukin-7 , Genetics , Severe Combined Immunodeficiency , GeneticsABSTRACT
<p><b>OBJECTIVE</b>This study aimed to investigate the possible role of Orphanin FQ (OFQ) in the regulation of hypo-thalamic gonadotropin-releasing hormone (GnRH) secretion.</p><p><b>METHODS</b>The method of push-pull perfusion and radioimmuno-assay (RIA) were adopted to examine the secretory profile of GnRH in the median eminence (ME) in freely moving ovari-ectomized (OVX) rats after intracerebroventricular (icv) injection of OFQ and/or [Nphe(1)]NC(1-13)NH(2) (NC13), a competitive antagonists of the opioid receptor-like 1 receptor (ORL1 receptor).</p><p><b>RESULTS</b>GnRH release from ME significantly decreased from 40 min to 80 min after the administration of 20 and 200 nmol OFQ in OVX rats (P < 0.05). This inhibitory effect of 20 nmol OFQ could be abolished by pretreatment with equal dose of NC13. More interestingly, GnRH secretion from ME was increased markedly 60 min after icv injection of 100 and 200 nmol NC13 (P < 0.05).</p><p><b>CONCLUSION</b>Our results suggested central administration of OFQ could inhibit the release of GnRH in the ME of hypothalamus through ORL1 receptor, providing further in vivo evidence supporting the role of OFQ in the control of GnRH secretion.</p>
Subject(s)
Animals , Female , Rats , Analysis of Variance , Dose-Response Relationship, Drug , Gonadotropin-Releasing Hormone , Metabolism , Median Eminence , Metabolism , Narcotic Antagonists , Opioid Peptides , Pharmacology , Ovariectomy , Methods , Peptide Fragments , Pharmacology , Radioimmunoassay , Rats, Sprague-Dawley , Receptors, Opioid , Metabolism , Secretory Pathway , Vasodilator Agents , Pharmacology , Wakefulness , PhysiologyABSTRACT
<p><b>AIM AND METHODS</b>To investigate the role of modulation by angiotensin AT1 receptor in sodium and water excretion induced by cholinergic agonist carbachol. Tyrosine hydroxylase immunoreactivity (TH-IR) in hypothalamus were also observed.</p><p><b>RESULTS</b>The natriuretic and diuretic effect induced by carbachol (CBC) were partially inhibited by pretreatment of losartan, a specific blocker of angiotensin AT1 receptor (P < 0.05). Immunohistochemistry showed that both TH-IR density and number of TH-IR positive neurons were markedly increased in PaPo, Arc, Pe and AHP of hypothalamus at 40 min after carbachol administration, as compared with NS group (P < 0.05). However, in losartan pretreated group, the number and the density of TH-IR were significantly decreased in such nuclei mentioned above except PaPo.</p><p><b>CONCLUSION</b>The results above suggest that brain AT1 receptor appears to be involved in mediating natriuresis induced by cholinergic stimulus. The blockade of AT1 receptor may down regulate the excitability of adrenergic neurons in Arc, Pe and AHP induced by CBC. We postulate that brain adrenergic and angiotensinergic pathway get involved in natriuresis induced by brain cholinergic stimulus together. Moreover, angiotensinergic neurons may influence the activity of adrenergic neurons in hypothalamus.</p>