Objective To analyze the incidence of BK virus (BKV) infection after kidney transplantation in our center and to evaluate the efficacy and safety of conversion treatment with Mizoribine (MZR) on BKV infection after kidney transplantation.Methods The information of recipients who received BK virus screening in hospital or outpatient during 2015-02 to 2016-12 in our center was retrospectively analyzed.The recipients positive for BKV were divided into experiment group (given conversion treatment with MZR) and control group (not given MZR conversion) according to the inclusion criteria.The negative rate of BKV,AR,hyperuricemia and the function of renal allograft during the conversion treatment with MZR were observed.Results 182 recipients accepted BKV screening during 2015-02 to 2016-12 and 68 cases were positive.The positive rate of BKV was 38.5 ％.The positive rate of peripheral blood specimens and midstream urine specimens was 7.1％ and 36.8％ respectively.Twelve recipients were positive for BKV in both peripheral blood specimens and midstream urine specimens.There were 27 recipients in experiment group and 36 cases in control group.Fourteen recipients positive for BKV became negative after MZR conversion in experiment group and the negative rate was up to 51.9％.The mean time of negative rate was 3.2 ± 2.7 (1-10) months after MZR conversion.During the conversion treatment with MZR,AR occurred in 1 case and was reversed by the impact therapy with Thymoglobulin in experiment group.The value of serum uric acid was maintained stable before and after MZR conversion under the action of uric-acidlowering drug.The renal function was kept stable in both experiment group and control group after renal transplantation.No deaths and renal allograft failure cases occurred in both groups during the research period.The 2-year survival rate for patients and kidneys was both 100％.Conclusion The incidence of BKV infection after kidney transplantation was high and the treatment scheme of MZR conversion was safe and effective.

Antrodia cinnamomea, an edible and medicinal fungus with significant economic value and application prospects, is rich in terpenoids, benzenoids, lignans, polysaccharides, and benzoquinone, succinic and maleic derivatives. In this study, the transcriptome of A. cinnamomea cultured on the wood substrates of Cinnamomum glanduliferum (YZM), C. camphora (XZM), and C. kanehirae (NZM) was sequenced using the high-throughput sequencing technology Illumina HiSeq 2000, and the data were assembled by de novo strategy to obtain 78,729 Unigenes with an N50 of 4,463 bp. Compared with public databases, about 11,435, 6,947, and 5,994 Unigenes were annotated to the Non-Redundant (NR), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genome (KEGG), respectively. The comprehensive analysis of the mycelium terpene biosynthesis-related genes in A. cinnamomea revealed that the expression of acetyl-CoA acetyltransferase (AACT), acyl-CoA dehydrogenase (MCAD), 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA), mevalonate pyrophosphate decarboxylase (MVD), and isopentenyl diphosphate isomerase (IDI) was significantly higher on NZM compared to the other two wood substrates. Similarly, the expression of geranylgeranyltransferase (GGT) was significantly higher on YZM compared to NZM and XZM, and the expression of farnesyl transferase (FTase) was significantly higher on XZM. Furthermore, the expressions of 2,3-oxidized squalene cyclase (OCS), squalene synthase (SQS), and squalene epoxidase (SE) were significantly higher on NZM. Overall, this study provides a potential approach to explore the molecular regulation mechanism of terpenoid biosynthesis in A. cinnamomea.

OBJECTIVETo investigate the accumulation of advanced glycation end products (AGE) and the inflammatory response of skin and wound in diabetic patients, and to analyze their relationship in vitro.

METHODSHistological staining and immunohistochemical staining was respectively performed on skin and wound tissue specimens collected from 10 patients with Type II diabetes mellitus (diabetes group) and 12 non-diabetic patients with skin injury (control group) to observe the arrangement of collagen and the distribution of inflammatory cells, and to determine the expression levels of AGE and its receptor (RAGE). Malondialdehyde (MDA) levels in skin and wound tissue homogenates were assayed by enzyme-linked immunosorbent assay. In vitro, human neutrophils were isolated and treated with RPMI-1640 culture medium or that containing AGE-human serum albumin in the concentration of 0.315, 0.625, 1.250 mg/mL, and they were identified as normal control (NC) group, low concentration (L) group, moderate concentration (M) group, and high concentration (H) group. Cell viability in each group was determined by MTT colorimetric assay, and the reactive oxygen species (ROS) in cell was measured with 2', 7'-dichlorofluorescein-diacetate. Data were processed with t test.

RESULTSCompared with those of skin in control group, collagens of skin tissues in diabetes group atrophied and disorderly arranged. Inflammatory cells in wounds in diabetes group were dispersed, in which collagens arranged loosely and irregularly, as compared with those of wounds in control group. Expression levels of AGE and RAGE of skin in diabetes group were higher than those in control group. In diabetes and control groups, especially in diabetes group, the numbers of RAGE-positive cells in wound tissue were more than those in skin tissue. Large amount of inflammatory cells with positive expression of RAGE were observed in diabetes group. MDA level of skin and wound tissue in diabetes group was respectively (6.3 ± 1.0), (7.1 ± 2.4) nmol per milligram protein, which were obviously higher than those in control group [(2.9 ± 1.0), (3.6 ± 1.4) nmol per milligram protein, with t value respectively 8.017, 4.349, P < 0.05 or P < 0.01]. Cell viability and ROS levels in neutrophils were increased in L, M, and H groups [(59 ± 8)%, (77 ± 5)%, (67 ± 6)% and 1.67 ± 0.14, 2.13 ± 0.17, 3.48 ± 0.48] as compared with those in NC group [(34 ± 5)% and 0.58 ± 0.06, with t value respectively 7.195, 14.890, 11.130 and 20.195, 24.905, 16.864, P < 0.05 or P < 0.01].

CONCLUSIONSAbnormal oxidative stress in diabetic skin leads to an atypical origin of wound repair. AGE-RAGE effect is a critical mediator for oxidative stress in diabetic wound tissue during wound healing.

Aged ; Case-Control Studies ; Diabetes Mellitus, Type 2 ; metabolism ; pathology ; Female ; Glycation End Products, Advanced ; metabolism ; Humans ; Male ; Middle Aged ; Oxidative Stress ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Serum Albumin ; metabolism ; Serum Albumin, Human ; Skin ; metabolism ; pathology ; Wound Healing

BACKGROUNDCyclosporin A (CsA) is a substrate of both cytochrome P450 3A (CYP3A) and P-glycoprotein (P-gp), some of the single nucleotide polymorphisms (SNPs) in these genes are associated with interindividual variations in CsA pharmacokinetics. We studied the influence of these SNPs on the incidence of rejection and CsA nephrotoxicity, as well as pneumonia within one year after renal transplant and post-transplantation diabetes mellitus (PTDM), in order to find whether genetic evaluation may help to identify patients at risk and to modulate CsA therapy to optimize graft and patient outcomes.

METHODSA total of 208 renal transplant recipients receiving CsA were genotyped for ABCB1 (C1236T, G2677T/A, and C3435T), CYP3A4 1G, and CYP3A5 3 by direct sequencing method. Retrospective case control study was utilized to identify the association between CYP3A4 1G, CYP3A5 3, ABCB1 genetic polymorphisms and CsA-related outcomes.

RESULTSThe patients with a CYP3A4 1G/ 1G genotype were found to have a higher incidence of acute rejection compared with those with CYP3A4 1/1.

CONCLUSIONCYP3A4 1G/1G genotype predict increased risk of acute rejection, so genetic evaluation may partly help to identify patients at risk and to modulate CsA therapy to optimize graft and patient outcomes.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Cyclosporine ; adverse effects ; therapeutic use ; Cytochrome P-450 CYP3A ; genetics ; Genotype ; Humans ; Immunosuppressive Agents ; adverse effects ; therapeutic use ; Kidney Transplantation ; Polymorphism, Genetic ; genetics ; Retrospective Studies

Astragalus polysaccharide(APS), one of the main active components of Astragali Radix, plays an anti-tumor effect by regulating the inflammatory microenvironment of tumors. Exosomes are small extracellular vesicles with a diameter ranging from 50 to 200 nm and carry several biological components from parental cells such as nucleic acids and proteins. When combined with recipient cells, they play an important role in intercellular communication and immune response. In this study, exosomes released from H460 cells at the inflammatory state or with APS addition activated by Toll-like receptor 4(TLR4) were extracted by ultracentrifugation and characterized by Western blot, transmission electron microscopy, and nanoparticle tracking analysis. The exosomal proteins derived from H460 cells in the three groups were further analyzed by label-free proteomics, and 897, 800, and 911 proteins were identified in the three groups(Con, LPS, and APS groups), 88% of which belonged to the ExoCarta exosome protein database. Difference statistical analysis showed that the expression of 111 proteins was changed in the LPS group and the APS group(P<0.05). The biological information analysis of the differential proteins was carried out. The molecular functions, biological processes, and signaling pathways related to the differential proteins mainly involved viral processes, protein binding, and bacterial invasion of proteasome and epithelial cells. Key differential proteins mainly included plasminogen activator inhibitor-1, laminin α5, laminin α1, and CD44, indicating that tumor cells underwent systemic changes in different states and were reflected in exosomes in the inflammatory microenvironment. The analysis results also suggested that APS might affect the inflammatory microenvironment through the TLR4/MyD88/NF-κB signaling pathway or the regulation of the extracellular matrix. This study is conducive to a better understanding of the mechanism of tumor development in the inflammatory state and the exploration of the anti-inflammatory effect of APS at the exosome level.
Humans ; Exosomes/metabolism* ; Proteomics ; Toll-Like Receptor 4/metabolism* ; Lipopolysaccharides ; Astragalus Plant/chemistry* ; Lung Neoplasms/metabolism* ; Polysaccharides/metabolism* ; Tumor Microenvironment

AIMTo investigate the influence of particle size and methoxypolyethyleneglycol (MePEG) molecular weight on the in vitro macrophage uptake and in vivo long circulating of recombinant human tumor necrosis factor-alpha (rHuTNF-alpha)-loaded stealth nanoparticles in rats.

METHODSThree sizes (approximately 80, 70 and 240 nm) of poly (methoxypolyethyleneglycol cyanoacrylate-co-n-hexadecyl cyanoacrylate) (PEG-PHDCA) nanoparticles loading rHuTNF-alpha were prepared at different MePEG molecular weights (Mr 2,000, 5,000, 10,000) using the double emulsion method. The in vitro macrophage uptake and in vivo long circulating properties in rats were examined and compared.

RESULTSThe uptake by macrophages decreased and the half-life of rHuTNF-alpha in rat increased with the increase of MePEG molecular weight or the decrease of particle size. The linear-ships between particle size and MePEG molecular weight and the in vitro macrophage uptake and in vivo long circulating properties were fairly good. Having the highest MePEG surface density (1.32 nm(-2)) , the shortest average distance between neighboring MePEG chain (0.87 nm) and the thicker fixed aqueous layer thickness (FALT, 5.16 nm), PEG5,000-PHDCA nanoparticles (80.0 nm) earned the strongest potency of decreasing uptake by macrophages and prolonging the half-life of rHuTNF-alpha in rat.

CONCLUSIONWithin the experimental limits, particle size and MePEG molecular weight had dramatic influence on in vitro macrophage uptake and in vivo long circulating properties of rHuTNF-alpha-loaded stealth nanoparticles.

Animals ; Cyanoacrylates ; chemistry ; Drug Carriers ; chemistry ; Drug Delivery Systems ; Macrophages ; physiology ; Male ; Mice ; Molecular Weight ; Nanoparticles ; Particle Size ; Phagocytosis ; drug effects ; Polyethylene Glycols ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; pharmacokinetics ; Tumor Necrosis Factor-alpha ; administration & dosage ; pharmacokinetics

In order to evaluate the efficiency of ITS2 and psbA-trnH sequences used as DNA barcodes to distinguish Plantaginis Semen from its adulterants, we collected 71 samples of Plantaginis Semen and its adulterants. The ITS2 and psbA-trnH sequences were aligned through Clustal W, and the genetic distances were calculated by kimura 2-parameter (K2P) model and the Neighbor-Joining (NJ) phylogenetic trees were constructed using MEGA 5.1. The results indicated that the ITS2 sequence lengths of Plantago asiatica and P. depressa were 199 bp and 200 bp, respectively; the maximum intra-specific K2P distance were lower than the minimum inter-specific K2P distance; the NJ tree based on ITS2 sequence indicated that Plantaginis Semen and its adulterants could be distinguished clearly. The sequence lengths of psbA-trnH of both P. asiatica and P. depressa were 340 bp; the maximum intra-specific K2P distances were lower than the minimum inter-specific K2P distance; the NJ tree based on psbA-trnH sequence showed that Plantaginis Semen can be distinguished clearly from its adulterants except for P. major. Therefore, ITS2 sequences can be used as an ideal DNA barcode to distinguish Plantaginis Semen from its adulterants.
Base Sequence ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; genetics ; Plantago ; classification ; genetics ; Quality Control ; Seeds ; classification ; genetics

In order to investigate the role of non-adrenergic non-cholinergic nerves in regulating mechanical and electrical activity of gastric circular smooth muscle, the effects of ATP and its analogues on gastric motility and electrical activities were observed in guinea-pig. In organ bath system, isometric force of the circular smooth muscle of guinea-pig gastric antrum was measured. Electrical activity of the muscle was recorded by using intracellular microelectrode. Electrical and mechanical activities were recorded by chart recorder. ATP and 2-MeSATP potentiated the mechanical activity but did not affect electrical activity in gastric circular smooth muscle. ATP and 2-MeSATP-induced contraction was effectively blocked by nonselective P2y-purinoceptor antagonist, reactive-blue-2 and suramin, but ATP-induced contraction was not blocked by alpha,beta-MeATP-induced desensitization of P2x-purinoceptors. However, alpha,beta-MeATP, P2x-purinoceptor agonist, attenuated slow waves with membrane hyperpolarization and inhibited contraction. The relaxation by beta,gamma-MeATP was blocked by alpha,beta-MeATP-induced desensitization of P2x-purinoceptors. ATP-induced contraction was blocked by external calcium-free, but not by nicardipine, a L-type calcium channel blocker. Indomethacin, a nonselective cyclooxygenase inhibitor, did not block ATP-induced contraction. The results suggest that: (1) ATP- and analogues-induced contraction is mediated by P2y-purinoceptor, whereas alpha,beta-MeATP-induced relaxation by P2x-purinoceptor in guinea-pig gastric antral circular smooth muscle. (2) ATP-induced contraction is dependent on extracellular calcium, but Ca2+ entry is not mediated by L-type calcium channel. (3) Prostaglandins are not involved in ATP- and analogue-induced contraction of gastric circular smooth muscle in guinea-pigs, and alpha,beta-MeATP-induced relaxation is related to membrane hyperpolarization.
Adenosine Triphosphate ; analogs & derivatives ; pharmacology ; Animals ; Electrophysiology ; Guinea Pigs ; In Vitro Techniques ; Microelectrodes ; Muscle Contraction ; drug effects ; physiology ; Muscle, Smooth ; drug effects ; physiology ; Purinergic Agonists ; Pyloric Antrum ; drug effects ; physiology ; Thionucleotides ; pharmacology

Acute Disease ; Glucocorticoids ; Graft Rejection ; Heart Rate ; Humans ; Hyperglycemic Hyperosmolar Nonketotic Coma ; Liver Transplantation

OBJECTIVETo evaluate the effects of indoor and outdoor PM2.5 (fine particulate matter, particulate matter with an aerodynamic diameter ≤ 2.5 µm) on lung function of college students in autumn and winter in Wuhan.

METHODSIn this panel study, 37 college students (excluded subject of respiratory disease and smoking history) aged 19 - 21 were investigated by cluster sampling in a university in Wuhan. The follow-up study lasted for 28 days in total, including two study periods, Oct. 29 to Nov. 11, 2009 (autumn) and Dec. 23, 2009 to Jan.5, 2010 (winter), the peak expiratory flow (PEF) of the college students were measured daily in the morning and evening in the university. PM10 and PM2.5 were monitored indoors and outdoors. The effects of PM on lung function of college students were analyzed by using generalized estimating equation (GEE).

RESULTSAverage daily concentrations of indoor, outdoor PM2.5 in autumn were (91.3 ± 43.7) and (104.2 ± 49.4) µg/m(3) respectively, while in winter the concentrations of indoor and outdoor PM2.5 were (110.6 ± 42.3) and (143.5 ± 51.2) µg/m(3). The single pollutant model showed that in winter, the evening PEF decrement was significantly associated with increasing outdoor PM2.5. With an increase of 10 µg/m(3) outdoor PM2.5, the PEF measured in the evening decreased 1.27 L/min (95%CI: 0.02 - 2.52 L/min, respectively). Meanwhile, the results showed that 2-days lagged outdoor PM2.5 was also significantly associated with morning PEF. An increase of 10 µg/m(3) 2-days lagged outdoor PM2.5 caused the decrease of 1.82 L/min (95%CI: -3.53 - -0.11 L/min) of PEF measured in the morning. Controlling the influence of gaseous pollutants and building the two pollutants models, the results indicated that no significant changes of PEF of students being exposed to PM2.5 on same day (lag 0) were observed. However, under consideration of SO2 effect, significant association between an increase of 10 µg/m(3) 2-days lagged outdoor PM2.5 and changes of morning PEF (-1.81 L/min, 95%CI: -3.51 - -0.11 L/min, P = 0.037) was found. The relationship between changes of concentrations and PEF was not observed in autumn in this study.

CONCLUSIONIn our panel study, exposure to outdoor PM2.5 is significantly associated with PEF among college students in winter, but not in autumn.

Air Pollutants ; China ; epidemiology ; Environmental Exposure ; Female ; Humans ; Male ; Maximal Midexpiratory Flow Rate ; Particulate Matter ; Respiratory Function Tests ; Seasons ; Students ; Young Adult