OBJECTIVEWe aimed to construct a recombinant adenovirus Ad-APN-EGFP and to evaluate the transfection efficiency and effect of the adenovirus promoting osteogenesis around implants.

METHODSRecombinant adenovirus Ad-APN-EGFP containing adiponectin (APN) was constructed by DNA sequencing and restriction enzyme digestion. Rat bone marrow stem cells (BMSCs) were transfected in vitro. Transfection efficiency and APN mRNA expression were tested. Animal models of rat femoral epiphysis and hydroxyapatite (HA)-coated implants were established. The Ad-APN-EGFP adenovirus at 10 μL was injected into the defect around HA-coated implants in the experimental group, whereas the same volume of phosphate-buffered saline was injected into the defect in the control group. Osteogenesis around implants was observed by hematoxylin-eosin staining four weeks after implantation.

RESULTSAd-APN-EGFP was successfully constructed by DNA sequencing and restriction enzyme digestion. The transfection efficiency of Ad-APN-EGFP to BMSCs was up to ≥ 90%. APN mRNA expression of BMSCs transfected with Ad-APN-EGFP was higher than that of the control group. Osteogenesis in the experimental group was more evident than that in the control group in vivo.

CONCLUSIONAd-APN-EGFP could be transfected into BMSCs and express APN mRNA at a high level. Ad-APN-EGFP could improve osteogenesis around implants in vivo.

Adenoviridae ; Animals ; Dental Implants ; Durapatite ; Osteogenesis ; Rats ; Transfection

AIM: To explore the predisposing factors, population characteristics and clinical features of severe fungal keratitis.
METHODS:The data of 233 cases 233 eyes of severe fungal keratitis in my hospital from January, 2008 to November, 2013 was retrospectively reviewed. The predisposing factors, population characteristics and clinical features were analyzed.
RESULTS: In 233 cases of severe fungal keratitis, the number of male patients was 153 ( 65. 7%) and the number ratio of male to female was 1. 9:1. The average age of them was (52. 7±11. 3), and most of them were middle-aged and elderly people living in the rural area (78. 1%) and were farmers ( 66. 1%) with low literacy (59. 7%). In 233 cases, 188 cases (80. 7%) possessed a clear history of ocular trauma, mainly caused by plant-based trauma (60. 9%). 90 cases (57. 3%) were infected with Fusarium, and 47 cases ( 29. 9%) by Aspergillus. The main treatment of severe fungal keratitis was surgery (87. 9%). 83 cases ( 52. 9%) were treated with penetrating keratoplasty, and in Fusarium and Aspergillus infected patients with severe fungal keratitis, 58. 4% ( 80/137 ) were performed with penetrating keratoplasty. In addition, patients treated with eye enucleation or evisceration, 68. 4% (13/19) were infected with Fusarium species.
CONCLUSION: Patients with severe fungalkeratitis in our hospital are mainly elderly male farmers living in rural, because of low economic condition and poor diagnosis consciousness. The main pathogens are Fusarium and Aspergillus species, and the major treatment is penetrating keratoplasty. Most of patients with poor clinical outcomes are infected with Fusarium species.

BACKGROUNDOur previous studies demonstrated that mutant IkappaBalpha (IkappaBalphaM) inhibited the occurrence, growth and angiogenesis of human glioblastoma multiform (GBM). However, the specific mechanism by which IkappaBalphaM regulates protein-degrading enzymes secreted from GBM to inhibit invasion and metastasis has remained unclear. The aim of the present study was to investigate the regulatory role and significance of IkappaBalphaM genes in the expression of tissue inhibitor of metalloproteinase (TIMP)-2 and matrix metalloproteinase (MMP)-9 in human GBM.

METHODSWe established the following four GBM cell lines stably expressing IkappaBalphaM by plasmid construction, gene transfection and screening for IkappaBalphaM protein expression: mutant IkappaBalpha-transfected cells (G36Delta-M), wild-type IkappaBalpha-transfected cells (G36Delta-W), empty plasmid transfected cells (G36Delta-P) and untransfected cells (G36Delta). The TIMP-2 and MMP-9 expression was detected by RT-PCR and Western blotting. Tumor cells were then implanted subcutaneously into nude mice to establish an animal model of ectopic tumor growth, and TIMP-2 and MMP-9 expression was determined by immunohistochemical methods.

RESULTSThe results showed that there was a significant increase in TIMP-2 expression and a significant decrease in MMP-9 expression in the G36Delta-M group at both the RNA and protein levels compared with the G36Delta-W group, G36Delta-P group and G36Delta group. Similar results were observed in the immunohistochemical staining analysis of tumor tissues. In the G36Delta-M group, TIMP-2 expression was significantly higher while MMP-9 expression was significantly lower than in the other three groups.

CONCLUSIONSOur findings indicate that IkappaBalphaM inhibits the activation of NF-kappaB. It significantly up-regulates TIMP-2 expression in human malignant glioma cells and down-regulates the expression of MMP-9. Thus, IkappaBalphaM maintains the integrity of the extracellular matrix and further inhibits the growth and metastasis of tumor tissues.

Animals ; Blotting, Western ; Cell Line, Tumor ; Glioblastoma ; genetics ; metabolism ; Humans ; I-kappa B Proteins ; genetics ; physiology ; Immunohistochemistry ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Mice ; Mice, Nude ; NF-KappaB Inhibitor alpha ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo establish a new visual educational system of virtual reality for clinical dentistry based on world wide web (WWW) webpage in order to provide more three-dimensional multimedia resources to dental students and an online three-dimensional consulting system for patients.

METHODSBased on computer graphics and three-dimensional webpage technologies, the software of 3Dsmax and Webmax were adopted in the system development. In the Windows environment, the architecture of whole system was established step by step, including three-dimensional model construction, three-dimensional scene setup, transplanting three-dimensional scene into webpage, reediting the virtual scene, realization of interactions within the webpage, initial test, and necessary adjustment.

RESULTSFive cases of three-dimensional interactive webpage for clinical dentistry were completed. The three-dimensional interactive webpage could be accessible through web browser on personal computer, and users could interact with the webpage through rotating, panning and zooming the virtual scene.

CONCLUSIONSIt is technically feasible to implement the visual educational system of virtual reality for clinical dentistry based on WWW webpage. Information related to clinical dentistry can be transmitted properly, visually and interactively through three-dimensional webpage.

Computer-Assisted Instruction ; methods ; Internet ; Oral Medicine ; education ; Software

To investigate the angiotensin I-converting enzyme (ACE) inhibitory activity of beta-chain hemoglobin fragments, 17 fragments were synthesized by microwave-assisted solid-phase synthesis method. Wang resin or Trt(2-Cl) resin, Fmoc and HBTU-HOBt were used as solid carrier, N-terminal amino acid protecting groups and coupling reagents, respectively. The ACE inhibitory, alpha-glucosidase inhibitory, antibacterial and antitumor activities of the synthesized fragments were assayed. In vitro, Val-Val-Tyr-Pro-Trp-Thr showed high ACE inhibitory activity (IC50 = 7.42 micromol x L(-1)). The results indicate that there are two active sites in Val-Val-Tyr-Pro-Trp-Thr-Gln-Arg-Phe, one consists of Val-Val-, and the other -Gln-Arg-Phe. Peptides showed high ACE inhibitory activity when the N-terminal was hydrophobic amino acid such as Val and C-terminal tripeptide contained Phe, Trp or Arg. Some of the fragments showed low a-glucosidase inhibitory activity. No antibacterial activity or antitumor activity was detected in vitro. The results indicate that these peptides have a potential antihypertensive effect and possible application in the treatment of hypertension.
Amino Acid Sequence ; Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Antihypertensive Agents ; pharmacology ; Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Glycoside Hydrolase Inhibitors ; Humans ; Peptide Fragments ; chemical synthesis ; chemistry ; pharmacology ; Peptidyl-Dipeptidase A ; drug effects ; Solid-Phase Synthesis Techniques ; methods ; alpha-Glucosidases ; drug effects ; beta-Globins ; chemical synthesis ; chemistry ; pharmacology

Adult ; Brain Neoplasms ; diagnosis ; secondary ; Female ; Humans ; Lymphomatoid Granulomatosis ; complications ; diagnosis ; Male ; Young Adult

Objective To investigate the distribution and antimicrobial resistance of Pseudomonas aeruginosa (P.aeruginosa) from intensive care units(ICUs) and general wards of a hospital,and provide scientific basis for rational use of antimicrobial agents in clinic.Methods Identification and antimicrobial susceptibility testing of clinically isolated bacteria in this hospital in 2016 were performed by VITEK 2 Compact automatic microbial analysis system,difference in antimicrobial resistance of P.aeruginosa between ICUs and general wards was compared.Results The tested specimens were mainly sputum in both ICUs and general wards,accounting for 78.7％ and 66.5％ respectively.There was no significant difference in the isolation rate of P.aeruginosa between ICUs and general wards (11.7％ vs 11.0％,P＞0.05).P.aeruginosa isolated from ICUs had the highest resistance rate to aztreonam (73.8％),resistance rates to piperacillin/tazobactam,cefoperazone/sulbactam,ceftazidime,imipenem,and meropenem were all up to more than 50％;P.aeruginosa detected in general wards had the highest resistance rate to aztreonam(59.6 ％),followed by piperacillin/tazobactam and imipenem,accounting for 48.0 ％ and 44.3 ％ respectively;resistance rates of P.aeruginosa isolated from ICUs to 12 kinds of antimicrobial agents were all higher thanthose of general wards(P＜0.05).Conclusion Resistance rate of P.aeruginosa from ICUs is higher than that in general wards,which should be paid attention,antimicrobial agents should be selected for clinical treatment of infection according to the results of antimicrobial susceptibility testing result.

Objective We aimed to assess the association between urinary bisphenol A(BPA)concentrations and gestational age in pregnant women. Methods A total of 248 pregnant women were recruited from a maternal and child care hospital in Shanghai. A questionnaire survey was completed to collect socio-demographic information and spot urine samples were collected during pregnancy. Gas chromatography-tandem mass spectrometry(GC-MS/MS)was used to measure BPA concentrations in urine samples. Linear relationship between urinary BPA level and gestational age was assessed by using generalized additive models. Multivariate regression model was used to evaluate associations of prenatal BPA exposure with gestational age. Results BPA was detected in all the urine samples. Median value and geometric mean of urinary BPA levels were 0.85 μg/L and 1.21 μg/L, respectively. Linear relationship between urinary BPA concentration and gestational weeks was confirmed(non-linear P > 0.05). Positive association between urinary BPA level and gestational age was indicated(regression coefficient, β = 0.19;95%CI:0.04-0.35;P = 0.016). However, it was only observed in girls, stratified by sex of newborns(β = 0.18;95%CI:0.03-0.34;P = 0.020). After stratification by trimester, no significant association was found in the second or the third trimesters. Conclusion Pregnant women are extensively exposed to BPA. Urinary BPA exposure during pregnancy may extend gestational age, especially in girls.

Insulin-like growth factor binding-protein-7 (IGFBP7) was obtained from our previous colonic adenocarcinoma (CRC) and normal mucosa suppression subtraction hybridization (SSH) cDNA libraries. By RT-PCR and immunohistochemistry, we found that IGFBP7 was overexpressed in CRC tissue compared to normal tissue. However, our in vitro experiments performed in 10 CRC cell lines showed that IGFBP7 expressed only in SW480 and Caco2 cell lines, which implied an underlying reversible regulatory mechanism. Using methylation-specific PCR (MSP) and bisulfite sodium PCR (BSP), we found that its expression was associated with DNA hypomethylation of exon1. This was further supported by the in vitro study which showed restored IGFBP7 expression after demethylation agent 5-aza-2'-deoxycytidine treatment. Correlation analysis between IGFBP7 expression and prognosis indicated that overexpression of IGFBP7 in CRC tissue correlated with favourable survival. Investigation of the functional role of IGFBP7 through transfection studies showed that IGFBP7 protein could inhibit growth rate, decrease colony formation activity, and induce apoptosis in RKO and SW620 cells, suggesting it a potential tumor suppressor protein in colorectal carcinogenesis. In conclusion, our study clearly demonstrated that IGFBP7 plays a potential tumor suppressor role against colorectal carcinogenesis and its expression is associated with DNA hypomethylation of exon 1.
Adenocarcinoma ; genetics ; metabolism ; Apoptosis ; genetics ; Cell Line, Tumor ; Colorectal Neoplasms ; genetics ; metabolism ; DNA Methylation ; Exons ; Gene Expression Regulation, Neoplastic ; genetics ; Humans ; Insulin-Like Growth Factor Binding Proteins ; genetics ; metabolism ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism

Adult ; Aged ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Meningeal Neoplasms ; pathology ; surgery ; Meningioma ; pathology ; surgery ; Middle Aged ; Postoperative Care ; methods ; Tomography, X-Ray Computed ; Treatment Outcome