The PDB culture medium was selected to ferment the endophyte strain, and the secondary metabolites of endophytic fungi Penicillium polonicum were studied. Combined application of Sephadex LH-20, ODS and HPLC chromatographies over the ethyl acetate extract of the fermented culture led to the isolation of 6 compounds. By spectral methods, the structures were elucidated as [3, 5-dihydroxy-2-(7-hydroxy-octanoyl)]-ethylphenylacetate (1), (3, 5-dihydroxy-2- octanoyl)-ethyl phenylacetate (2), (5, 7-di- hydroxy-9-heptyl)-isobenzo pyran-3-one (3), 3-(hydroxymethyl) 4-(1E)-1- propen-1-yl-(1R, 2S, 5R, 6S)-7-oxabicyclo [4.1.0] hept-3-ene-2, 5-diol (4), (E)-2-methoxy-3-(prop-1-enyl) phenol (5) and p-hydroxylphenylethanol (6).
Biological Factors ; chemistry ; metabolism ; Endophytes ; chemistry ; isolation & purification ; metabolism ; Fabaceae ; microbiology ; Fermentation ; Penicillium ; chemistry ; isolation & purification ; metabolism ; Secondary Metabolism

Objective To evaluate in vivo tracking of swine mesenchymal stem cells (MSCs) la-beled with super paramagnetic iron oxide (SPIO) in intraportal transplantation by a clinical 1.5T MR.Methods MSCs were isolated from swine and cultured as well as expanded, which were then incuba-ted with SPIO (Feridex I. V.). Prussian blue staining was performed for showing intracelluar irons.To establish a swine model of acute liver necrosis, 0.5 g/kg of D-galactosamine was administrated to 10 pigs. MSCs(labeled cells in six, unlabeled cells in four)were injected into liver via portal veins. MR imaging was performed with a clinical 1.5T MR immediately before and at 6 h, 3 d, 7 d, 14 d after transplantation, respectively. Results Prussian blue staining of SPIO labeled MSCs could be effec-tively labeled and the labeling efficiency was almost 100%. Signal intensity loss in liver by SPIO labe-ling on FFE sequence persisted until 14 days after transplantation. Histological analysis by Prussian blue staining showed homing of labeled MSCs in liver after 14 days, primarily distributing in hepatic sinusoids and liver parenchyma. Conclusion MSCs can be labeled with SPIO in vitro successfully.MRI can monitor magnetically labeled MSCs transplanted into liver.

Cholesteatoma, Middle Ear ; Humans ; Male ; Young Adult

Objective To investigate the effects of maternal long-term exposure to low-dose trichlorfon on the serum paraoxonase (PON) activity of pregnant mice and development of embryos. Methods Female ICR mice (n=120) were randomly divided into control group and trichlorfon groups of different doses,and were managed by intragastric injection with trichlorfon of 0,2,10 and 50mg/kg,respectively. All the mice were managed once a day for a consecutive of 27 days,and were subjected to mating. The pregnant mice were continued to be managed with trichlorfon for 3 days,and were sacrificed on day 3 of gestation. The serum PON and acetylcholinesterase (AchE) activities were detected,and the development of embryos was evaluated. Results The serum PON activity of 2,10 and 50mg/kg trichlorfon group were (14.15?1.22),(12.78?1.80) and (10.45?1.95)IU/mL,respectively,and that of 50mg/kg trichlorfon group was significantly lower than that of control group [(13.37?2.31)IU/mL] (P0.05),while the the percentage of abnormal embryos of 50mg/kg trichlorfon group had an increased tendency. Conclusion Long-term exposure to low-dose trichlorfon can inhibit serum PON and AchE activity in pregnant mice without obvious effect on the development of embryos.

Objective To investigate the dialectical nursing effect of SHI's Bianstone comprehensive therapy on the patients with cervical vertigo.Methods 76 patients with cervical vertigo from Conghua Chinese Medicine Hospital accepted the dialectical nursing during treatment of SHI's Bianstone comprehensive therapy.Results After treatment,36 of 76 patients with cervical vertigo were cured,which accounted for 47％,28 patients had some tangible results,accounting for 37％,9 patients had effective results,accounting for 12％,and 3 patients were ineffective,accounting for 4％.The total effective rate was 96％.Conclusions SHI' s Bianstone comprehensive therapy is very helpful for treating patients with cervical vertigo,and dialectical nursing can improve the treatment effectiveness.

Objective To explore the effect of CD40 small interfering RNA(siRNA)on the expressions of pe-ripheral blood interleukin(IL)-21 and IL -35 in rats with experimental autoimmune myocarditis (EAM)and its sig-nificance.Methods Twenty 6 -8 week male Lewis rats were divided into normal group,EAMgroup,CD40 siRNA group and siRNA group by using random number table,with 5 rats in each group.The normal rats were induced with phos-phate buffer saline in double foot pads on day 0 and day 7,while the rest 3 groups were induced with cardiac myosin protein to establish EAMmodels.The rats in CD40 siRNA group and siRNA group were respectively injected with CD40 siRNA and siRNA slow virus expression vector through the tail vein of rats on day 7.The rats were executed on 21 day after echocardiogram examination was made.The histopathologic changes were observed by using light microscope and the myocardial histopathology scores were calculated.Enzyme -linked immunosorbent assay was used to determine the levels of IL -21 and IL -35 in peripheral blood.Results (1)Except the normal group,the total incidence rate of rats of each group was 100%,and there was no rat death.(2)Compared with EAM group,the heart mass/body ratio and myocardial histopathology scores were lower in CD40 siRNA group,and the differences were significant (3.13 ±0.21 vs 3.80 ±0.29,2.22 ±0.43 vs 3.32 ±0.51,F =0.332,0.456,all P <0.05).(3)The echocardiogram showed that there was only 1 rat in EAM group with massive pericardial effusion,and there was no pericardial effusion in CD40 siRNA group.EAMgroup,CD40 siRNA group and siRNA group displayed hypertrophy of the ventricular septum and left ventricular wall,narrow heart cavity and weakening of ventricular wall motion.The left ventricular shortening rate in CD40 siRNA group was significantly higher than that in the EAMgroup[(63.34 ±11.06)% vs (38.56 ±6.98)%,F =16.080,P <0.05].(4)The peripheral blood level of IL -21 in CD40 siRNA group was lower than that in EAM group [(141.19 ±17.46)ng/L vs (157.81 ±17.58)ng/L,F =57.008,P <0.05],while its level of IL -35 was signifi-cantly higher than that in the EAMgroup [(195.96 ±18.26)ng/L vs (174.78 ±13.91 )ng/L,F =31.727,P <0.05].(5)The level of IL -21 in peripheral blood was positively correlated with myocardial histopathology scores in EAM group (r = 0.69,P < 0.05 ),but IL -35 was negatively correlated with myocardial histopathology scores (r =-0.64,P <0.05).Conclusions CD40 siRNA might relieve the myocardial inflammation and reduce the myocar-dial injury of EAMrats.The levels of IL -21 and IL -35 can partly reflect the degree of myocardial injury.The mecha-nism may be related to down -regulating the expression IL -21 and up -regulating the expression of IL -35.

Animals ; Apolipoproteins E ; deficiency ; genetics ; Blood Vessels ; anatomy & histology ; Fluorescence ; Lipofuscin ; Mice ; Mice, Knockout

Objective: To analyze diagnosis rate of chronic kidney disease (CKD) in hospitalized pediatric patients in a single center and understand pediatricians′ awareness of CKD. Methods: This was a cross-sectional study. Children who were admitted to the Division of Pediatric Nephrology, Peking University First Hospital from January 1, 2008 to December 31, 2017 and met the diagnostic criteria of CKD (kidney disease: improving global outcomes 2012 guideline) were recruited. A total of 4 472 cases were enrolled. Original CKD diagnosis was collected from the home page of medical records. Actual CKD diagnosis was validated and corrected by reviewing medical records and recalculating glomerular filtration rate. The diagnosis rate and influencing factors of pediatric CKD, the distribution and etiology of actual CKD were analyzed. The comparison between groups were performed with χ² test. Results: In 4 472 cases, there were 3 470 cases in actual CKD stage 1, among which only 24 cases were in original CKD stage 1. There were 543 cases in actual CKD stage 2-3, among which only 181 cases were in original CKD stage 2-3. Three hundred and one cases were in actual CKD stage 4-5, including 290 cases in original CKD stage 4-5. In addition, there were 43 cases with unknown CKD stage and 115 cases with acute kidney injury. Compared to original CKD diagnosis, the diagnosis rates of CKD stage 1-5 were 0.7% (24/3 470), 16.7% (58/348), 63.1% (123/195), 90.7% (78/86) and 98.6% (212/215), respectively. The proportions of actual CKD stage 1-5 were 80.4% (3 470/4 314), 8.1% (348/4 314), 4.5% (195/4 314), 2.0% (86/4 314) and 5.0% (215/4 314). The etiology of actual CKD included primary glomerular disease (62.2%, 2 686/4 314), secondary glomerular disease (19.7%, 849/4 314), hereditary kidney disease (9.1%, 391/4 314), congenital abnormalities of the kidney and urinary tract (CAKUT) (3.1%, 135/4 314), tubulointerstitial disease (2.2%, 94/4 314) and etiology uncertain (2.1%, 89/4 314). The leading cause of end stage renal disease was etiology uncertain (31.1%, 67/215), followed by hereditary kidney disease (24.2%, 52/215), CAKUT (16.3%, 35/215) and primary glomerular disease (16.3%, 35/215). Conclusions Among actual CKD hospitalized pediatric patients, the diagnosis rate of CKD given by physicians at discharge was relatively low, especially patients in earlier CKD stages, which reflected serious lack of physicians′ awareness of CKD.

OBJECTIVECongenital nephrotic syndrome (CNS) is defined as heavy proteinuria or nephrotic syndrome occurring before 3 months of age. It is characterized by early onset, resistance to steroid therapy and progressing to end-stage renal disease (ESRD). In recent years, several genes associated with CNS have been identified, such as NPHS1, NPHS2 and WT1. The mutations of these genes have been identified in the patients with CNS in Finland, other European countries, North Africa, North America, and Asia, respectively. However, the investigation of the above genes has not been performed in Chinese CNS patients. In this study, NPHS1 mutations in a Chinese family with CNS were detected and analyzed.

METHODSThere were two CNS patients in the investigated family. The proband, a 45-day-old boy, was born at fullterm and weighed 2700 g at birth. The placenta weighed 450 g. At the age of 10 days, generalized edema, proteinuria, hypoproteinemia, and hypoalbuminemia were found without renal insufficiency. The proband's sister, with the same phenotype and normal renal function, underwent renal biopsy at 5 years of age. Their parents and elder half-sister all had normal phenotypes. Genomic DNA samples were extracted from peripheral bloods of the proband, his family members and 50 unrelated, normal individuals. All 29 exons and exon-intron boundaries of NPHS1 were detected in the proband by polymerase chain reaction (PCR), direct DNA sequencing, and restriction enzyme analysis.

RESULTSThree heterozygous mutations of NPHS1, namely, G928A (D310N), 1893-1900del 8 (CGAAACCG), and G2869C (V957L) were identified in the proband. These mutations involved exons 8, 14, and 21. The same genotype was found in the proband's sister who had the same phenotype, but was not detected in proband's elder half-sister who had normal phenotype. Fifty normal individuals had no these mutations. The proband's mother with normal urinalysis had G928A (D310N) heterozygous mutation, and the father with normal urinalysis had two heterozygous mutations of 1893-1900del 8 (CGAAACCG) and G2869C (V957L). At the same time, three types of single nucleotide polymorphisms (SNPs), E117K (rs3814995), S1105S (rs2071327), and IVS27+45c > t, were confirmed in the proband. Another variant, IVS8+68 a > g had also been found.

CONCLUSIONThis is the first report about NPHS1 mutations in Chinese CNS kindred. These three heterozygous mutations of NPHS1 are novel genetic defects of CNS, which have not been described before.

Asian Continental Ancestry Group ; genetics ; Exons ; Genotype ; Humans ; Infant ; Male ; Membrane Proteins ; genetics ; Mutation ; Nephrotic Syndrome ; congenital ; genetics ; Pedigree ; Phenotype ; Polymerase Chain Reaction ; Sequence Analysis, DNA

AIMPulmonary hypertension is a common complication of congenital heart disease with a left-to right shunt. The mechanism of pulmonary hypertension induced by high pulmonary blood flow is still not fully understood. Recent studies showed that hydrogen sulfide (H2S) could relax vascular smooth muscle cells. But the change of the system of H2S in pulmonary hypertension induced by high pulmonary blood flow was not reported. We studied the influence on expression of CSE mRNA and production of hydrogen sulfide in rat lung tissues by L-Arginine, in order to demonstrate a regulating role of nitric oxide (NO) in the regulation of cystathionine-gamma-lyase/hydrogen sulfide system (CSE/H2S).

METHODSThirty male SD rats were randomly divided into shunting group, shunting with L-Arginine group, and control group. Abdominal aorta and inferior vena cava shunting was produced in rats of the later group. Pulmonary artery mean pressure (mPAP) and the hypertrophy of right ventricle of each rat were analyzed. The expression of lung tissue CSE mRNA was measured using quantitative reverse transcription-polymerase chain reaction and in situ hybridization. The activity of CSE in lung tissue was measured according to chemical analysis.

RESULTSmPAP was significantly increased in shunted rats compared with normal control (P < 0.01), the expression of lung tissue CSE mRNA and the activity of CSE in lung tissue were decreased in shunt group (P < 0.01). However, L-arginine significantly attenuated pulmonary artery pressure, but augmented the expression of lung tissue CSE mRNA as well as the activity of CSE in lung tissue.

CONCLUSIONL-Arginine reverses the down-regulation of CSE/H2S system in high pulmonary blood flow-induced pulmonary hypertension.

Animals ; Arginine ; metabolism ; Cystathionine gamma-Lyase ; genetics ; metabolism ; Gene Expression ; Gene Expression Regulation ; Hydrogen Sulfide ; metabolism ; Lung ; blood supply ; Male ; Nitric Oxide ; metabolism ; Rats ; Rats, Sprague-Dawley