BACKGROUND: Guided tissues regeneration (GTR) has been successfully used in the treatment of periodontal diseases by using biocompatible membrane to prevent tooth epithelial cell growing towards root and to facilitate the formation of a certain gap which is favorable for periodontal membrane cells to occupy the surface of the root, thereby to rebuild the peridental structure. OBJECTIVE: To verify whether GTR can promote the repair of periodontal bone defect and the formation of new bones and whether combined use of GTR membrane, artificial bone, and growth factors can acquire much more new periodontal tissue. DESIGN, TIME AND SETTING: An animal observation experiment was performed at the laboratory of Medical College of Dalian University between February and July 2006. MATERIALS: Basic fibroblast growth factor (bFGF) freeze-dry powder (50 mg/ampoule) was thoroughly dissolved with ddH2O to prepare 5 g/L bFGF solution, bFGF solution was dropped onto collagen membranes with a size of 10 mm ×10 mm (5 mg bFGFs per piece of collagen membrane). METHODS: Cementum and the alvelor bone 6 mm below the cement-enamel junction were removed to induce periodontal defect in 16 rabbits with big ears. Artificial bone with bFGF and GTR membrane were used to fold around the defect on the right side, followed by reduction and suture, serving as experimental side. The left side underwent identical experimental procedure, with the exception of application of artificial bone with bFGF and GTR membrane, serving as control side. MAIN OUTCOME MEASURES: Carrier material absorption, inflammation of inner connective tissues, and status of junctional epithelium were examined through the use of microscope. RESULTS: The control side exhibited no concrescence, while the expedmental side displayed concrescence of the cementum to various extents. CONCLUSION: Application of GTR membrane, artificial bone, and bFGF can promote the concrescence of damaging bone.

We wished to study the intracellular transport of adenoviruses. We constructed a novel recombinant adenovirus in which the structural protein IX was labeled with a mini-singlet oxygen generator (miniSOG). The miniSOG gene was synthesized by overlapping extension polymerase chain reaction (PCR), cloned to the pcDNA3 vector, and expressed in 293 cells. Activation of miniSOG generated sufficient numbers of singlet oxygen molecules to catalyze polymerization of diaminobenzidine into an osmiophilic reaction product resolvable by transmission electron microscopy (TEM). To construct miniSOG-labelled recombinant adenoviruses, the miniSOG gene was subcloned downstream of the IX gene in a pShuttle plasmid. Adenoviral plasmid pAd5-IXSOG was generated by homologous recombination of the modified shuttle plasmid (pShuttle-IXSOG) with the backbone plasmid (pAdeasy-1) in the BJ5183 strain of Eschericia coli. Adenovirus HAdV-5-IXSOG was rescued by transfection of 293 cells with the linearized pAd5-IXSOG. After propagation, virions were purified using the CsC1 ultracentrifugation method. Finally, HAdV-5-IXSOG in 2.0 mL with a particle titer of 6 x 1011 vp/mL was obtained. Morphology of HAdV-5-IXSOG was verified by TEM. Fusion of IX with the miniSOG gene was confirmed by PCR. In conclusion, miniSOG-labeled recombinant adenoviruses were constructed, which could be valuable tools for virus tracking by TEM.
Adenoviruses, Human ; chemistry ; genetics ; metabolism ; Arabidopsis Proteins ; chemistry ; genetics ; metabolism ; Flavoproteins ; chemistry ; genetics ; metabolism ; Humans ; Phototropins ; chemistry ; genetics ; metabolism ; Singlet Oxygen ; chemistry ; Staining and Labeling ; Transfection

OBJECTIVE:To investigate the toxicity effect of Jianpi shengxue granule on perinatal rats. METHODS:Based on body mass,pregnant rats were randomly divided into negative control group and Jianpi shengxue granule low-dose,medium-dose, high-dose groups(0.77,2.31,6.93 g/kg),21 in each group,and intragastrically given relevant medicines from the 15th d of preg-nancy until 21st d after delivery,once a day. Effects of Jianpi shengxue granule on general toxicity and fertility of maternal rats were observed,as well as the effects on athletic ability,learning and memory ability,fertility of F1 offspring and early viability of F2 offspring. RESULTS:In terms of toxicity effect on maternal rats,compared with control group,the days of pregnancy in Jianpi shengxue granule high-dose group was significantly prolonged(P<0.05),initial body mass of administration was significantly re-duced (P<0.01). In terms of toxicity effect on offspring,compared with negative control group,the initial body mass of F1 off-spring in Jianpi shengxue granule high-dose group was significantly reduced(P<0.05),the time of 4th and 5th time for founding underwater platform was significantly shortened(P<0.01),and there was no obvious effect on fertility of F1 offspring and early vi-ability of F2 offspring (P>0.05);there was no obvious effect on the indexes of offspring in Jianpi shengxue granule medium-dose,low-dose groups. CONCLUSIONS:The no toxic dose of Jianpi shengxue granule in maternal and offspring rats is 2.31 g/kg.

Objective To investigate the clinical application value of the levels of heparin-binding protein (HBP) in cerebrospinal fluid (CSF) for intracranial infectious diseases. Methods A case-control study was conducted. 150 patients after craniotomy(73 in the postoperative bacterial intracranial infection group, 77 in the postoperative non-infection group) admitted to the Department of Neurology of the People's Hospital of Liaoning Province from December 2016 to May 2018 were collected. At the same time, 46 patients without operation (14 in the non-bacterial intracranial infection group, 32 patients without intracranial infection were selected as control group whose white blood cell count (WBC) values in CSF were all below 10 × 106/L) in the same period were also collected. According to the diagnostic criteria for severe intracranial infection, the patients with bacterial intracranial infection were divided into 26 cases of mild intracranial infection group and 47 cases of severe intracranial infection group. According to the Glasgow Outcome Scale (GOS) score at the time of discharge, the patients were divided into 30 cases of good prognosis group (GOS score 4-5 points) and 43 cases of poor prognosis group (GOS score 1-3 points). The concentrations of HBP in CSF were tested with latex immunoturbidimetry, and the concentrations of procalcitonin(PCT) in cerebrospinal fluid and serum were tested with electrochemiluminescence, and cerebrospinal fluid routine were tested with instrument method, and the concentrations of total protein(TP) in cerebrospinal fluid were tested with turbidimetry. The differences of the laboratory test indicators in each group were statistically analyzed, and the levels of HBP in CSF of patients with different degrees of intracranial infection and different prognosis were compared. Comparison of two independent samples was performed using the Mann-Whitney U test. Results The HBP levels in cerebrospinal fluid were 187.00 (73.00, 635.00) ng/ml, 10.00 (3.50, 32.00) ng/ml, 1.50 (0, 4.00) ng/ml, 3.00 (1.00, 4.00) ng/ml in post-craniotomy bacterial intracranial infection group, uninfected group after craniotomy, non-bacterial intracranial infection group and control group respectively. The cerebrospinal fluid levels of WBC count were 1280.00 (363.00, 4327.00)×106/L, 63.00 (18.50, 300.00)×106/L, 5.00 (3.00, 14.75)×106/L, 3.00 (2.00, 5.75)×106/L. The absolute value of cerebrospinal fluid neutrophils were 1216.00 (225.50, 3895.50)×106/L, 24.00 (2.00, 209.50)×106/L, 1.00 (1.00, 3.00)×106/L, 1.00 (1.00, 1.00)×106/L. The cerebrospinal fluid levels of PCT were 0.16 (0.10, 0.32) ng/ml, 0.09 (0.07, 0.14) ng/ml, 0.07 (0.06, 0.12) ng/ml, 0.07 (0.06, 0.13) ng/ml. The serum levels of PCT were 0.36 (0.15, 1.09) ng/ml, 0.09 (0.04, 0.16) ng/ml, 0.08 (0.04, 0.13) ng/ml, 0.07 (0.03, 0.11) ng/ml. The levels of HBP, WBC, neutrophils, PCT in CSF and serum PCT in the post-craniotomy bacterial intracranial infection group were significantly higher than those in the uninfected group after craniotomy (Z=-9.246,-6.759,-6.741,-4.477,-6.202, P<0.05), non-bacterial intracranial infection group(Z=-5.840,-5.412,-5.259,-2.923,-5.104,P<0.05) and the control group (Z=-7.905,-7.919,-7.335,-4.397,-5.474, P<0.05). There were significant differences in the levels of HBP, WBC and neutrophils in CSF(Z=-3.763,-3.444,-3.041,P<0.05) and no significant differences in CSF and serum PCT (Z=- 0.869, - 1.850, P>0.05)between the uninfected group after craniotomy and the non-bacterial intracranial infection group. There were significant differences in the levels of HBP, WBC and neutrophils in CSF(Z=-4.496,-6.685,-4.842,P<0.05) and no significant differences in CSF and serum PCT(Z=-0.676,-1.303, P>0.05)between the uninfected group after craniotomy and the control group. There were no significant differences in the levels of HBP, PCT in CSF and serum PCT (Z=-0.861,-0.514,-0.273, P>0.05)and significant differences in the levels of WBC and neutrophils in CSF(Z=-2.756,-3.060, P<0.05) between the non-bacterial intracranial infection group and the control group. The levels of HBP in CSF in the severe intracranial infection group were significantly higher than those in the mild intracranial infection group(Z=-6.267, P<0.05). The levels of HBP in CSF in the poor prognosis group were significantly higher than those in the good prognosis group(Z=-7.064, P<0.05). The area under the ROC curve for the diagnosis of bacterial intracranial infection by HBP, WBC, neutrophils, TP, PCT in CSF and PCT in serum was 0.986, 0.987, 0.945, 0.945, 0.770 and 0.914, respectively. The area under the ROC curve for differential diagnosis of bacterial intracranial infection and non-bacterial intracranial infection was 0.994, 0.958, 0.961, 0.929, 0.747 and 0.936, respectively. Conclusions HBP in CSF is an ideal indicator for the diagnosis of bacterial intracranial infection. It is important to distinguish between bacterial intracranial infection and non-bacterial intracranial infection. The extent of increase is related to the severity of infection and prognosis of the disease.

Objective To explore the application of nanodisc in functional and drug discovery research of G protein-coupled receptor (GPCR).Methods The purified recombinant 5-Hydroxytryptamine 2B receptor (5-HT2BR) was reconstituted into nanodisc complex.Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and size exclution chromatography were performed to evaluate the reconstitution reaction,followed by the use of surface plasmon resonance to validate the ligand-binding activity of 5-HT2BR after reconstitution.Results 5-HT2B R was effectively self-assembled into nanodisc while maintained its binding activity toward the antagonist SB204741.Conclusions The presented study provided potential application of 5-HT2B R-nanodisc for the development of subtype-selective drugs against 5-HT2B R and the fundamental of utilizing nanodisc for GPCR structural and functional studies as well as drug discovery.

Objective:To explore the influencing factors of neutrophil to lymphocyte ratio (NLR) in patients with chronic heart failure (CHF) and the relationship between NLR and cardiac contractile function.Methods:From September 2016 to May 2018, 135 CHF patients were enrolled in the cardiovascular department of Fenyang hospital, Shanxi Province for prospective research.According to the level of NLR, the patients were divided into three groups: low NLR group (<2.3, 46 cases), middle NLR group (≥2.3~≤4.3, 45 cases), high NLR group (>4.3, 44 cases). The basic clinical data, laboratory examination data and noninvasive cardiac hemodynamic indexes of the three groups were compared.Results:(1) By comparing the data of different NLR groups, we found that there were significant differences in (N-terminal pro-brain natriuretic pepfide, NT-proBNP)( F=4.485, P=0.013), total bilirubin( F=6.085, P=0.003), albumin( F=3.695, P=0.027). (2)NLR was correlated with NT proBNP, total bilirubin, albumin( r=0.267, 0.256, -0.243, P=0.002, 0.003, 0.005, respectively). (3)Multiple linear regression analysis showed that NLR of CHF patients was correlated with left ventricular ejection fraction (LVEF), NT proBNP, total bilirubin and albumin (standard regression coefficients were -0.239, 0.223, 0.247 and -0.213, respectively, P<0.05). (4) Pearson correlation analysis showed that NLR and cardiac output ( r=-0.173, P=0.045), cardiac index ( r=-0.175, P=0.042), LVEF ( r=-0.278, P=0.001), maximum ejection velocity ( r=-0.207, P=0.016), systolic index ( r=-0.214, P=0.013), hearther index(HI) ( r=-0.179, P=0.038), cardiac work (CW) ( r=-0.235, P=0.006), cardiac workindex (CWI)=( r=-0.244, P=0.004) were negatively correlated. Conclusion:NT-proBNP, total bilirubin, albumin and LVEF are the factors that affect the NLR of patients with CHF.NLR has a certain value in the evaluation of CHF, therapeutic effect and prognosis prediction.

Bacterial Typing Techniques ; China ; Genotype ; Humans ; Mycobacterium tuberculosis ; Tuberculosis

Objective:To explore the effect of the location and size of region of interest (ROI) on the measurement of liver fat by means of quantitative computed tomography (QCT).Methods:A total of 98 subjects who were examined with QCT for bone mineral density examination from December 25, 2019 to January 17, 2020 were recruited continuously from the Department of Health Management of Henan Provincial People′s Hospital. The liver fat content was measured by QCT workstation. The ROI was located respectively in the left lobe, the right anterior lobe and the right posterior lobe of the liver, and it was measured independently by the A measurer and B measurer. The central position of the ROI was fixed and the diameter was increased, and it was measured by the A measurer. In this study, Friedman test was used to compare the differences of measurement results in different positions or sizes of ROI, and intra-class correlation coefficients (ICC) was used to evaluate the repeatability of inter-measurers.Results:There was a significant difference for liver fat content under different positions of ROI (χ2=62.306, P<0.001), but no difference under different seizes of ROI (χ2=1.088, P=0.581). The ICC values of the inter-measurers repeatability analysis of the A measurer and B measurer in the left lobe, right anterior lobe and right posterior lobe of the liver were 0.847, 0.917 and 0.874, all more than 0.75, and the reproducibility was good. Conclusions:When QCT technique is applied to the measurement of liver fat content, the location conditions of ROI may affect results, so it is necessary to select multiple ROI in the whole liver for measurement. The inter-measurers repeatability of QCT in different parts of the liver is good.

Objective:To provide support for the clinical application of quantitative computed tomography (QCT) in the measurement of liver fat content, this study evaluated the intra-observer and inter-observer reproducibility of liver fat content measured by QCT in a population receiving physical examinations.Methods:From April to July 2019, 291 people were consecutively selected who underwent QCT examination in the health management department of Henan Provincial People′s Hospital. There were 214 males (73.5%) and 77 females (26.5%), aged 48.7±11.0. We measured liver fat content by QCT workstation. Three observers (A, B, C) measured their liver fat content independently, then observer A performed re-testing two weeks later. The mean value of the two measurements from observer A was taken as the final result. Measurement data were described by mean±SD. Intra-observer and inter-observer reproducibility were assessed using intra-class correlation coefficients ( ICC). Results:The first measurement result for observer A was 10.46±5.55 and the second measurement for observer A was 10.66±5.59, resulting in a final value of 10.56±5.51. The measurement results of observers B and C were 10.70±5.45 and 10.86±5.77, respectively. The ICC value of liver fat content values measured by the three observers was 0.960 (95% CI: 0.951-0.967, P<0.001) and the ICC value of liver fat content values for the two measurements of observer A was 0.953 (95% CI: 0.941-0.962, P<0.001). The ICC values were>0.75, so reproducibility of results was good. Conclusions:If the measurement method is consistent, the results for liver fat content measured by a conventional CT scanner and QCT workstation will have good reproducibility between and within observers, and will also have certain clinical application prospects.

Objective To develop an indicator cell line for detection of human cytomegalovirus (HCMV) based on inducible expression of green fluorescent protein (GFP) gene.Methods GFP reporter gene and the promoter of HCMV UL54 gene (UL54p) were amplified by PCR method;GFP gene was used to replace the luc2 coding sequence in pGL4.17 [luc2/Neo] vector,and then UL54p was inserted into the multiple cloning sites (MCS) to generate pGL4UL54p-GFP.pGL4UL54p-GFP plasmid was transfected into MRC-5T,a telomerase reverse transcriptase (TERT) gene-immortalized MRC-5 cell line,by mixing with lipofectamine 3000 reagent.G418-resistant cell colonies were isolated and subjected into screening by observing the inducible expression of GFP under fluorescence microscope after HCMV infection.The specificity to HCMV of the obtained cell line was tested through infection of human adenovirus type 5 or influenza A virus subtype H1N1.Results The recombinant plasmid pGL4UL54p-GFP was successfully constructed.Nine G418-resistant cell colonies were obtained,and two of them could express GFP after HCMV infection among which MRC-5TUG#7 gave brighter fluorescence.No GFP-positive cells were seen after challenging MRC-5TUG#7 with adenovirus or influenza virus,suggesting an acceptable specificity.Conclusion MRC-5-based indicator cell line for HCMV detection was successfully established,which may be used for HCMV isolation and titration.