At present, obesity has become an independent risk factor for chronic kidney disease through its direct or indirect effects. Urine mitochondrial DNA (mtDNA) is a new marker of mitochondrial damage in various kidney diseases, but the differences in urinary mtDNA between obese patients and healthy subjects and the effect of obesity intervention on urinary mtDNA copy number are unknown. In January 2019, " The Journal of Clinical Endocrinology & Metabolism" published an article " Bariatric surgery reduces elevated urinary mitochondrial DNA copy number in obese patients" [Lee H, Oh S, Yang W, et al. J Clin Endocrinol Metab, 2019, 104(6): 2257-2266], with the permission of the original journal, we translate the article into Chinses. This article studied the difference of urinary mtDNA levels between obese patients and healthy subjects and the changes of urinary mtDNA after metabolic surgery intervention. The results revealed that obesity was associated with the increase of urinary nicotinamide adenine dinucleotide dehydrogenase subunit 1 (mtND-1) copy number, and urinary mtDNA copy number was elevated in obese patients. Metabolic surgery reduced urinary mtND-1 copy number in obese patients. This study shows that metabolic surgery can alleviate mitochondrial damage in kidney cells in obese patients.

AIM AND METHODSThe effects of losartan (after operation 2 week to 10 week, 5 mg/kg d ig) on generation of AT1R-AA in sera were observed during development of hypertension in rats. The renovascular hypertension (RVH) model was established by two-kidney one-clip method, a synthetic peptide corresponding to amino acid sequence 165-191 of the second extracellular loop of the angiotensin II-1 receptor (AT1R) was used as antigen, SA-ELISA were used to examine sera AT1R autoantibody (AT1R-AA).

RESULTSThe frequencies and titres of AT1R-AA after operation one week rats were significantly increased (P < 0.05). The treatment with losartan not only inhibited structural and functional changes, but also the frequencies and titres of AT1R-AA was significantly lower (P < 0.05) than RVH group.

CONCLUSIONIt is suggested that the losartan significantly inhibits generation of the AT1R-AA.

Animals ; Autoantibodies ; biosynthesis ; blood ; Hypertension, Renovascular ; blood ; immunology ; Losartan ; pharmacology ; Male ; Rats ; Rats, Wistar ; Receptors, Angiotensin ; immunology

This study was purposed to detect single nucleotide polymorphisms (SNP) of 2 pharmacokinetics-related genes in K562 and K562/A02 cell lines. Leukemia cell line K562 and its resistant line K562/A02 were cultured, the genomic DNA was isolated by QIAamp DNA Blood Mini kit, primers were designed, the related DNA fragments were amplified by PCR. The SNP genotyping of mthfr gene rs1801131, rs1801133 and rs2274976 and dpyd gene rs1801159, rs1801160 and rs17376848 was performed by means of matrix assisted laser desorption ionization-time of flight mass spectrometry method (MALDI-TOFMS). The results showed that the genotype of mthfr gene locus 1801131 was AC, rs1801133 was CC, rs2274976 was GG, genotype of dpyd gene locus 1801159 was GG, rs1801160 was GG, rs17376848 was AA in both K562 and K562/A02 cell lines. It is concluded that the above-mentioned loci of mthfr and dpyd genes in K562 and K562/A02 cell lines are not expressed differently.
DNA Mutational Analysis ; DNA Primers ; Dihydrouracil Dehydrogenase (NADP) ; genetics ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; Genotype ; Humans ; K562 Cells ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Polymorphism, Single Nucleotide

PURPOSETo investigate the in vitro effect of short interfering RNAs (siRNAs) against Nogo receptor (NgR) on neurite outgrowth under an inhibitory substrate of central nervous system (CNS) myelin.

METHODSThree siRNA sequences against NgR were designed and transfected into cerebellar granule cells (CGCs) to screen for the most effcient sequence of NgR siRNA by using reverse transcription polymerase chain reaction (RT-PCR) and immunofluorescence staining. NgR siRNA sequence 1 was found the most efficient which was then transfected into the CGCs grown on CNS myelin substrate to observe its disinhibition for neurite outgrowth.

RESULTSCompared with the scrambled control sequence of siRNA, the NgR siRNA sequence 1 significantly decreased NgR mRNA level at 24 h and 48 h (p <0.05), which was recovered by 96 h after transfection. NgR immunoreactivity was also markedly reduced at 24 and 48 h after the transfection of siRNA sequence 1 compared with that before transfection (p<0.05). The NgR immunoreactivity was recovered after 72 h post-transfection. Moreover, the neurite outgrowth on the myelin substrate was greatly improved within 72 h after the transfection with siRNA sequence 1 compared with the scrambled sequence-transfected group or non-transfected group (p<0.05).

CONCLUSIONsiRNA-mediated knockdown of NgR expression contributes to neurite outgrowth in vitro.

Animals ; Cells, Cultured ; Myelin Sheath ; physiology ; Neuronal Outgrowth ; physiology ; Nogo Receptor 1 ; antagonists & inhibitors ; genetics ; physiology ; RNA, Small Interfering ; Rats ; Rats, Sprague-Dawley

Natural product is an important source of new drug research and development (R&D). Traditional Chinese medicine (TCM) innovation is the key step for its modernization and internationalization. However, due to the complexity of TCM, there are many difficulties and confusions in this process. Target-based drug discovery is the mainstream model and method of R&D. TTD, short for therapeutic target database, is developed by National University of Singapore. Besides a large amount of information on drug targets, the database also contains considerable information related to natural products. This paper briefly introduces the TTD, analyzes the natural products derived drugs/compounds recorded in TTD, which we think might provide some inspiration for the innovation of TCM.
Databases, Factual ; Drug Delivery Systems ; Drug Discovery ; Drugs, Chinese Herbal ; Medicine, Chinese Traditional ; Singapore

BACKGROUNDIt is very important for the clinical management to test for minor HIV-1 resistance mutations accurately and sensitively. The conventional genotypic assays of HIV drug resistance detection based on sequencing can only discriminate the mutations which present in more than 20% - 30%. The aim of this study was to evaluate allele-specific real-time PCR (ASPCR) to detect the resistance-related mutations located at positions 103, 184 and 215.

METHODSWe developed the allele-specific PCR assay, using the most common drug resistance mutations in Chinese AIDS patients, K103N, M184V/I, T215F/Y as a model system. The standards were constructed by cloning the wild-type and mutant DNA fragments into the T-vector. We designed specific primers to discriminate mutant templates in the real-time PCR using SYBR green as a fluorescence reporter. And then we evaluated the ASPCR assay and tested 140 clinical samples using this method.

RESULTSThe sensitivities of ASPCR assay were 0.04% for K103N, 0.30% for M184I, 0.40% for M184V, 0.03% for T215F and 0.02% for T215Y. The intra-assay and inter-assay coefficients of variation were less than 0.42. One hundred and forty plasma samples were tested by ASPCR and dynamic resistance curves of ten patients were obtained.

CONCLUSIONSDrug resistance emerged half a year after the start of antiretroviral therapy. The mutation of T215Y emerged 1 to 1.5 years after starting treatment and then increased rapidly. The ASPCR assay we developed was a sensitive, accurate and rapid method to detect the minor HIV-1 variants and it can provide earlier and more drug-resistance information for HIV research and AIDS antiretroviral therapy.

Alleles ; Drug Resistance, Viral ; HIV-1 ; drug effects ; genetics ; Humans ; Mutation ; Real-Time Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVETo investigate the influence of combined supplementation of glutamine (Gln) and recombinant human growth hormone (rhGH) on the protein metabolism in severely burned patients.

METHODSSixty severely burned patients were enrolled in the study and were randomly divided into control (C, n = 20) and Gln with rhGH (Gln + rhGH, n = 20) groups. The patients in C group received glycine as the placebo, while those in Gln group took Gln orally in dose of 0.5 g kg(-1) d(-1) during 1-14 postburn days (PBDs). For the patients in Gln + rhGH group rhGH was administered subcutaneously in dose of 0.2 U kg(-1) d(-1) in addition to glutamine in same dosage beginning on the 7 PBD for 7 days. The plasma Gln concentration in the 3 groups of patients was determined on the 1st, 7th and 14th PBD and the plasma albumin level was determined on 14th and 21st PBD. The wound healing rate of the patients within 30 PBSs and the total hospital stay days were recorded.

RESULTSThe plasma Gln concentration in Gln + rhGH group of patients was evidently higher than that in C group after 7 PBD[(452.28 +/- 21.72) micromol/L vs(325.12 +/- 25.34) micromol/L, P < 0.05]. The plasma albumin level in Gln + rhGH group was obviously higher than that in C and Gln groups on the 21st PBD (P < 0.05). The wound healing rate in Gln + rhGH group was evidently higher than that in Gln and C groups on the 30th PBD (P < 0.05). The total hospital stay days in Gln + rhGH group were obviously less than that in C and Gln groups (P < 0.05 or 0.01).

CONCLUSIONCombined administration of Gln and rhGH could be beneficial to the elevation of plasma Gln level in severely burned patients and the systemic protein synthesis was therefore enhanced and the wound healing rate was improved.

Adult ; Aged ; Burns ; metabolism ; therapy ; Female ; Glutamine ; administration & dosage ; blood ; therapeutic use ; Human Growth Hormone ; administration & dosage ; therapeutic use ; Humans ; Male ; Middle Aged ; Recombinant Proteins ; administration & dosage ; therapeutic use ; Treatment Outcome ; Wound Healing ; drug effects ; Young Adult

OBJECTIVETo identify non-invasive biomarkers for diagnosis and/or prognosis of liver fibrosis in chronic hepatitis B (CHB).

METHODSPeripheral blood samples were obtained from 48 patients with CHB, including 24 with mild fibrosis (stage 1, S1) and 24 with severe fibrosis (stage 4, S4), and subjected to Ficoll density gradient centrifugation in order to obtain enriched samples of peripheral blood mononuclear cells (PBMCs).The PBMC proteomes of the two groups were assessed by first separating the total proteins by two-dimensional gel electrophoresis (2DE) and then identifying the differentially expressed proteins by liquid chromatography combined with tandem mass spectrometry (LCMS/MS).

RESULTSThe enriched PBMC samples from the S1 group and the S4 group had similar amounts of platelets [(19.268+/- 6.413) * 109/L and(19.480+/- 6.538) * 109/L, respectively); however, for both, the platelet amounts were 5 to 15-fold lower than that of the normal reference (100-300 *109/L). There was no significant difference found between the platelet amounts in the S1 patients and healthy controls (P=0.930). Twelve differentially expressed proteins were identified through 2DE-LC-MS/MS, including proteins such as moesin and NADH dehydrogenase [ubiquinone] iron-sulfur protein 3 that are involved in various biological processes like cell movement, cell adhesion, kinase signaling and transcription.

CONCLUSIONs The 12 proteins with differential expression in S1 and S4 patients with CHB and liver fibrosis may represent markers related to development and/or progression of liver fibrosis.

Biomarkers ; Disease Progression ; Electrophoresis, Gel, Two-Dimensional ; Hepatitis B, Chronic ; complications ; Humans ; Leukocytes, Mononuclear ; chemistry ; metabolism ; Liver Cirrhosis ; etiology ; metabolism ; pathology ; Mass Spectrometry ; Prognosis ; Proteome ; Proteomics ; Tandem Mass Spectrometry

Objective To study the effect of arthroscopic internal fixation combined with arthrodesis on patients with advanced ankle arthritis and American Orthopedic Ankle Association Scoring System (AOFAS) and visual analogue scale (VAS). Methods 84 patients with advanced ankle arthritis from January 2012 to January 2015 were randomly divided into experimental group (42 cases) and control group (42 cases) by random number method. The patients in the control group were treated with traditional open ankle arthrodesis, the experimental group under the arthroscopic assisted internal fixation joint fusion. Then compare the time of surgery, intraoperative blood loss, postoperative hospitalization time and complication. The follow-up period was 12 to 36 months. Used the AOFAS score system to evaluate the curative effect. Use VAS to evaluate the degree of ankle pain. Results The operation time and intraoperative blood loss were significantly lower in the experimental group than that in the control group (P < 0.05). The postoperative hospital stay and the time of joint fusion were lower in the experimental group than that in the control group (P < 0.05). The incidence of complication (9.52%) in the experimental group was significantly lower than that in the control group (25.57%) (P < 0.05). The results of follow-up showed that the VAS and AOFAS scores of the experimental group were better than those in the control group (P < 0.05). Conclusion The procedure of arthroscopic endoscopic fusion is short, the bleeding rate is low, the incidence of complications is low, the healing rate is high, and the follow-up effect is accurate. It is suitable for clinical use.

OBJECTIVETo analyze the clinical effect of spinal cord decompression and lavage therapy on chronic cervical spinal cord injury and explore the possible mechanism.

METHODSFifty-seven patients with chronic cervical spinal cord injury treated in our hospital from January, 2008 to January, 2015 were enrolled, including 17 with multilevel cervical disc herniation, 25 with long segmental ossification of the posterior longitudinal ligament, 13 with hypertrophy or calcification of neck ligamentum flavum, and 2 with old cervical fractures. Open-door spinal canal laminoplasty via a posterior approach and decompression in simple extramedullary decompression was performed in 31 cases (group A), and open-door spinal cord incision decompression via a posterior approach, saline irrigation, and spinal canal laminoplasty in intramedullary decompression was performed in 26 cases (group B). The pre-operative cerebrospinal fluid in group B patients was collected to examine the inflammatory factors. All the patients were followed up and evaluated for pre- and postoperative JOA scores to calculate the improvement rate with regular examinations by X-ray, CT or MRI.

RESULTSImaging examinations 2 weeks after the operation showed obvious relief of the primary lesion in both groups, and the improvement of high signals was better in group B than in group A. The mean improvement rate at 12 months after the operation was 52.33% in group A and 61.52% in group B (P<0.05), and the mean JOA score was significantly higher in group B than in group A (14.80∓1.51 vs 13.58∓0.56; P<0.05). Cerebrospinal fluid leakage occurred in 3 cases, epidural hematoma in 2 cases, internal fixation loosening in 1 case in group A; portal shaft fracture and internal fixation loosening occurred in 1 case in group B. Postoperative recovery time was shorter in group B and entered the platform phase in 3 months. The inflammatory factors IFN-γ, IL-17F, IL-6 and sCD40L were all significantly higher than the normal levels after spinal cord injury, and the increment of IL-6 was the most conspicuous (P<0.05).

CONCLUSIONIntramedullary and extramedullary decompression can achieve better outcomes than extramedullary decompression in patients with chronic cervical cord injury. This may be related not only to relieving adhesions and secondary compression by cutting the dura under the microscope, but also to removal of local inflammatory factors.