Objective Through detecting CD4+CD25+ T regulatory cells(Treg)in the peripheral blood in children suffering autoimmune diseases and normal controls to learn about the changes of Tregs during the diseases and to acquire some references for clinical diagnosis and treatment.Methods The data were reviewed for CD4+CD25+ Treg cells of the 93 children diagnosed as pediatric autoimmune disease in Children′s Hospital Affiliated to Capital Institute of Pediatrics from Nov.2007 to Jun.2008.Thirty-five normal children in the contempora-neous physical examination were selected as the control group.The percentage of CD4+CD25+ Treg cells and CD4+ T cells to the total T cells were determined by flow cytometric method.Data of the JRA group(22 cases),SLE group(12 cases) and HSP group(12 cases),which were the first three according to the number of cases,were respectively compared with the controls.Independent-samples t test was performed for a statistic analysis with SPSS 11.5 software.Results 1.The percentages of CD4+CD25+ Treg cells to the total T cells and CD4+ T cells in the autoimmune diseases children[(6.14?3.21)% and(21.85?11.68)%,respectively] were both higher than those in the control group[(3.68?1.02)% and(12.83?3.61)%,respectively Pa

Inosine 5′-monophosphate dehydrogenase (IMPDH) is a key enzyme catalyzing the rate-limiting step of de novo nucleotide synthesis in vivo. In recent years, it has become a therapeutic target for anti-virus, anti-bacterial, anti-cancer, anti-parasitic and other diseases. IMPDH inhibitors have been shown to inhibit viral prolife-ration in host cells by depleting guanosine 5′-monophosphate (GMP), the raw material required for viral replication in host cells, with broad-spectrum antiviral properties. In order to find novel anti-coronavirus drugs, this study screened 22 potential IMPDH inhibitors from 70 000 natural small molecule libraries based on IMPDH protein structure using molecular docking and ROC calculation for virtual screening. With ribavirin as the positive control drug, Huh7 cell and H460 cell models were used to verify the anti-coronavirus HCoV-229E and HCoV-OC43 activities of 22 selected target compounds. Among them, compounds 11, 12, 15 and 16 showed inhibitory activity against coronavirus HCoV-229E. The compounds 4, 12, 13 and 15 showed inhibitory activities against coronavirus HCoV-OC43. 12 and 15 showed significant inhibitory activity against both two coronaviruses, and their efficacy was similar to ribavirin at the same dose, which can be further studied as a lead compound for IMPDH inhibitors.

OBJECTIVE: To select specific DNA aptamer for determining ketamine by FluMag-SELEX. METHODS: Based on magnetic beads with tosyl surface modification as solid carrier and ketamine as target, a random ssDNA library with total length of 78 bp in vitro was compounded. After 13 rounds screening, DNA cloning and sequencing were done. Primary and secondary, structures were analyzed. The affinity, specificity and Kd values of selected aptamer were measured by monitoring the fluorescence intensity. RESULTS: Two ssDNA aptamers (Apt#4 and Apt#8) were successfully selected with high and specific abilities to bind ketamine as target with Kd value of 0.59 and 0.66 μmol/L. The prediction of secondary structure was main stem-loop and G-tetramer. The stem was the basis of stability of aptamer's structure. And loop and G-tetramer was the key of specific binding of ketamine. CONCLUSION FluMag-SELEX can greatly improve the selection efficiency of the aptamer, obtain the ketamine-binding DNA aptamer, and develop a new method for rapid detection of ketamine.
Aptamers, Nucleotide/metabolism* ; DNA ; DNA, Single-Stranded/genetics* ; In Vitro Techniques ; Ketamine/metabolism* ; Oligonucleotides ; SELEX Aptamer Technique/methods*

Objective To identify the peptide that have strong ability binding to HCMV-UL149 encoded protein,and to analyze the characteristics of the amino acid sequence of UL149-binding peptides.Methods Expressed UL149 proteins of three genotypes were used to screen the binding peptide in the random peptide display library,then the encoding sequence of binding peptides in the selected clones were sequenced.The amino acid sequences of the binding peptides were analyzed for their homology,and were com pared with those of the known protein in protein banks.Results The homologous amino acid sequence W/A/F/V-D/E-D/E-G-W/F/I/L were found within the binding peptides selected by proteins of all the three UL149 genotypes proteins,and no difference between three groups was found.The alignment with amino acid sequences of the known proteins in protein banks showed that the binding peptides of UL149 putative protein have homologous amino acid sequences with immunoglobulin heavy chain variable region(IgHV),the serine/threonine protein kinases,compliment factor H,zinc finger protein,MHC Ⅰ molecule,eukaryotic translation initiation factor,nuclear factor and so on.Conclusion The UL149 encoding proteins have binding ability to proteins mentioned above,and might interfere with the immunity responds to HCMV infection through multiple mechanisms.

Objective To investigate the effects and mechanisms of pin1(peptidyl-prolyl isomerase 1) inhibitor ju?glone on migration and invasion in glioma cell line U-87 MG. Methods Glioma cells were treated with juglone at 0, 0.8, 1.6 and 3.2μmol·L-1. Wound-healing assay and invasion assay were performed to examine the inhibitory activity of ju?glone on glioma cell line U-87 MG. Western blot was used to analyze the protein expression of β-catenin, VEGF, MMP-2 and MMP-9. Results The wound-healing assay showed that the wound-healing rate in juglone-treated groups was 46.04%±6.25%and 30.05%±13.35%at concentrations of 1.6μmol·L-1 and 3.2μmol·L-1 , repectively. Juglone treat?ment significantly reduced the wound-healing rate compared with controls (P<0.05). Transwell invasion assay showed that the number of invaded cells in juglone–treated groups was 103.67 ± 5.69 and 77.33 ± 7.77 at the concentrations of 1.6μmol·L-1 and 3.2μmol·L-1 , respectively. Juglone treatment significantly reduced cell invasion compared with con?trols (P<0.05). Treatment with juglone significantly down-regulated the expression levels ofβ-catenin, VEGF, MMP-2 and MMP-9 in U-87 MG cells in a dose-dependent manner. Conclusion The present data suggests that juglone has a significant inhibitory action on cell migration and invasion through down-regulation of theβ-catenin and its downstream VEGF, MMP-2 and MMP-9 protein expressions in glioma cell line U-87 MG.

Following the wishes of volunteer patients,316 cases of type 2 diabetics of our community center in 2008 carried out a three-year intervention from 2008 to 2011 on the basis of rational drug therapy plus the interventions of health education,regular review and lifestyle strengthening.And regular monitoring and recording were performed on the parameters of body mass index (BMI),blood pressure,blood lipids,fasting glucose,2 h postprandial blood glucose,glycosylated hemoglobin (HbA1c) and liver & kidney function.After intervention,some indicators changed:BMI (25.2 ±3.5) vs.(25.0 ±3.3) kg/m2,systolic blood pressure (129.1 ± 11.8) vs.(126.2 ±7.9) mm Hg(1 mm Hg=0.133 kPa),fasting glucose (7.80 ±2.81) vs.(7.25 ± 1.96) mmol/L,2 h postprandial blood glucose (11.04 ±4.60) vs.(9.83 ±3.60) mmol/L,HbA1c (7.39 ± 1.61) vs.(7.17 ± 1.65)％,total cholesterol (5.08 ±1.21) vs.(4.74 ± 1.35) mmol/L,low density lipoprotein cholesterol(LDL) (3.09 ± 0.87) vs.(2.85 ±0.83) mmol/L,high density lipoprotein cholesterol (HDL) (1.27 ± 0.33) vs.(1.41 ± 0.32) mmol/L,serum creatinine 65 vs.72 μmol/L,uric acid 300 vs.317 μmol/L.And the differences were statistically significant (P ＜ 0.05).After intervention,blood pressure compliance rate increased from 72.5％ to 88.0％,LDL-C compliance rate improved from 27.2％ to 38.6％,HDL-C compliance rate of 54.9％ increased to 66.9％.And the differences were statistically significant (P ＜ 0.05).The pre-intervention combined compliance rate of 11.4％ (n =36) rose to 17.7％ (n =56).And there was significant difference (P =0.024).

To retrospectively analyze the clinical data of a child with congenital broncho-bile duct fistula(CBBF) in Guiyang Children′s Hospital in June 2019.A female, aged 7 years and 6 months old, patient presented cough with a large amount of yellow green mucus.The main clinical manifestation was recurrent pulmonary infection after birth.After the fistula was found by electronic bronchoscope, doctors cooperated with imaging department, anesthe-siology department and pediatric surgery department.After treatment, the child recovered and discharged.There are few reports on CBBF.This study suggested that, in view of the refractory pneumonia with recurrent pulmonary infection and yellow green sputum after birth, and that the effect of anti-infection treatment was poor, clinicians should pay attention to the CBBF, take bronchoscopy as soon as possible, and make early diagnosis by combining with imaging technology, thus formulating a reasonable diagnosis and treatment plan under multidisciplinary cooperation, so as to improve the diagnosis and treatment of this rare disease clinical diagnosis and treatment level, and reduce missed diagnosis and misdiagnosis as well.

Objective To evaluate the relationship between endometriosis fertility index (EFI) and pregnancies after laparoscopic surgery in endometriosis-associated infertility.Methods From Jan.2005 to Jan.2010,medical documents of 350 infertile patients due to endometriosis undergoing laparoscopic surgery were studied retrospectively.Pregnancy outcomes were followed up by telephone.EFI was calculated by history factors,least function score and some aspects of the revised American Fertility Society (r-AFS) endometriosis stage.The cumulative pregnancy rate was calculated and compared by Kaplan-Meier survival analysis.Results Within 3 years after surgery,the cumulative pregnancy rates among patients with EFI score 8,9,10 were 62.5％,69.8％ and 81.1％,respectively.There was no significant difference in pregnancy rates among those three groups of patients ( P =0.24 ).The cumulative pregnancy rates among patients with EFI score 5,6,7 were 49.8％,43.9％ and 41.6％,respectively,which did not reach statistical difference ( P =0.83 ).The cumulative pregnancy rates of EFI score 8 - 10 was significantly higher than that of EFI score 5 -7 (71.8％ vs.44.4％,P =0.000).The patients with EFI score 0 -4 was quite small with only 33 cases,among which 15 cases were pregnant.Conclusions There is relationship between EFI and pregnancy in patients with endometriosis-associated infertility.EFI is meaningful to guide post surgical treatment.

OBJECTIVEThe value of plasma brain natriuretic peptide (BNP) and N-terminal proBNP (NT-proBNP) can reflect cardiac function and therefore can be used for diagnosing congestive heart failure (CHF) and evaluating cardiac function. There are few reports, however, on the value of BNP and NT-proBNP in pediatric cases of congenital heart defect. The aim of this study was to assess the value of plasma NT-proBNP in the diagnosis of CHF and evaluation of cardiac function in pediatric patients with ventricular septal defect (VSD).

METHODSFifty-one patients with VSD aged from 2 months to 2 years old (mean 7.9 months) were enrolled. According to the modified Ross Score, the patients were divided into three groups, no CHF group (20 patients), mild CHF group (18 patients) and moderate to severe CHF group (13 patients). Fifteen age-matched normal children were used as controls. Plasma NT-proBNP was measured using enzyme immunoassay. All patients had complete echocardiographic study, including measurement of left ventricular end diastolic volume index (LVEDVI), left ventricular end systolic wall stress (LVSEWS), heart rate corrected mean velocity of circumferential fiber shortening (mVcFc), left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS), and contractility index (Con). The correlation between plasma NT-proBNP level and modified Ross Score and echocardiographic cardiac functional indexes was determined. The sensitivity, specificity and ROC curve of plasma NT-proBNP for diagnosing CHF was studied.

RESULTSPlasma NT-proBNP was positively correlated with modified Ross Score (r = 0.75, P < 0.01). Plasma NT-proBNP concentration in moderate to severe CHF group (2061 +/- 908) fmol/ml was significantly higher than that of mild CHF group (810 +/- 335) fmol/ml, and Plasma NT-proBNP concentration in mild CHF group was higher than that in no CHF group (309 +/- 68) fmol/ml. 97.14% of normal controls and subjects in no CHF group had their plasma NT-proBNP below 400 fmol/ml. 83.3% of children in mild CHF group had their plasma NT-proBNP between (400-1400) fmol/ml while in moderate and severe CHF group 84.6% of children had their plasma NT-proBNP beyond 1400 fmol/ml. Plasma NT-proBNP was also positively correlated with LVEDVI and LVSEWS. There was no correlation among mVcFc, LVEF, LVFS, Con and plasma NT-proBNP concentration. Using plasma NT-proBNP concentration > or = 400 fmol/ml as cut-point for diagnosing CHF, the sensitivity was 89.3%, the specificity was 91.2%, and the area under the ROC curve was 0.944.

CONCLUSIONSPlasma NT-proBNP level could be used to assess cardiac function and diagnose CHF in pediatric patients with VSD.

Echocardiography ; Female ; Heart Failure ; blood ; Heart Septal Defects, Ventricular ; blood ; diagnosis ; Humans ; Infant ; Male ; Natriuretic Peptide, Brain ; blood ; Peptide Fragments ; blood ; Ventricular Function, Left

OBJECTIVETo investigate the polymorphism of human cytomegalovirus (HCMV) UL150 open reading frame (ORF) in low-passaged clinical isolates, and to study the relationship between the polymorphism and different pathogenesis of congenital HCMV infection.

METHODSPCR was performed to amplify the entire HCMV UL150 ORF region of 29 clinical isolates, which had been proven containing detectable HCMV-DNA using fluorescence quantitative PCR. PCR amplification products were sequenced directly, and the data were analyzed.

RESULTSTotally 25 among 29 isolates were amplified, and 18 isolates were sequenced successfully. HCMV UL150 ORF sequences derived from congenitally infected infants were high variability. The UL150 ORF in all 18 clinical isolates shifted backward by 8 nucleotides leading to frame-shift, and contained a single nucleotide deletion at nucleotide position 226 compared with that of Toledo strain. The nucleotide diversity was 0.1% to 6.8% and the amino acid diversity was 0.2% to 19.2% related to Toledo strain. However, the nucleotide diversity was 0.1% to 6.4% and amino acid diversity was 0.2% to 8.3% by compared with Merlin strain. Compared with Toledo, 4 new cysteine residues and 13 additional posttranslational modification sites were observed in UL150 putative proteins of clinical isolates. Moreover, the UL150 putative protein contained an additional transmembrane helix at position of 4-17 amino acid related to Toledo.

CONCLUSIONHCMV UL150 ORF and deduced amino acid sequences of clinical strains are hypervariability. No obvious linkage between the polymorphism and different pathogenesis of congenital HCMV infection is found.

Amino Acid Sequence ; Base Sequence ; Cytomegalovirus ; genetics ; isolation & purification ; Cytomegalovirus Infections ; congenital ; virology ; DNA, Viral ; genetics ; Genes, Viral ; Genetic Variation ; Humans ; Infant ; Infant, Newborn ; Molecular Sequence Data ; Open Reading Frames ; Polymorphism, Single Nucleotide ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics