[Objective]To discuss bone marrow derived mesenchymal stem cells(MSCs) culture in vitro and the curative effect of repairing cancellous bone defects in rabbits with the compound of allogenic MSCs and fibrin.[Method]MSCs were isolated and culture-expanted from rabbit bone marrow.Cancellous bone defects were created on bilateral femur condylar in 15 Japanese big ear rabbits,the left bone defects were treated with allogenic MSCs and fibrin,the right were treated with fibrin only.The bone defects were examined radiologically and histoloyically at the 2nd,5th,8th week after operation.[Result]The defects with only fibrin implantation couldn't heal spontaneously and no bone formation were found at any time-point.In the defects with compound of MSCs and fibrin implantation,there were a few bone formation within 2 weeks.By the 5th week plentiful callus were found,at the end of the 8th week,the defects healed.There was no immune respone except a few inflammatory cells infiltrate.[Conclusion]The compound grafts of allogenic MSCs and fibrin shows low immunogenicity within 8 weeks and is an effective method for repairing cancellous bone defects.

OBJECTIVETo investigate the homology of Staphylococcus aureus (SA) strains isolated from nose and skin lesions of impetigo children.

METHODSTotally 263 outpatients aged 3 months to 14 years who were seen by the Department of Dermatology of Beijing Children's Hospital between August 2005 and March 2006 were enrolled in this study. The isolations from nose and skin lesions of 58 impetigo children who were randomly selected from these 263 children with spa sequence were typed. The sequence results of SA were analyzed using special websites.

RESULTSThere were 106 impetigo patients in these 263 children. The isolation rate of SA was 78.3% in the nose of 106 impetigo patients and was 21.0% in that of the rest 157 patients (P < 0.01). The age of all nasal carriers was concentrated in 1-6 years. Among the 106 impetigo patients, 30 patients had their primary lesions on the face (including 28 cases of SA nose isolates) and 76 patients had their primary lesions on the other parts of body (including 56 cases of SA nose isolates) (P < 0.01). The spa typing showed that 26 of the 30 impetigo patients had the same type pairs of nose and skin.

CONCLUSIONSSA isolated from the skin lesions and nose of impetigo patients has remarkably homology. Nasal carriage of SA may be closely relevant with the occurrence of impetigo.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Impetigo ; microbiology ; Infant ; Male ; Nasal Cavity ; microbiology ; Sequence Homology ; Skin ; microbiology ; Staphylococcus aureus ; classification ; genetics ; isolation & purification

OBJECTIVETo study serum ferritin and neuron-specific enolase (NSE) levels in children with hand-foot-mouth disease (HFMD) complicated by acute viral encephalitis and their clinical significance.

METHODSSerum levels of ferritin and NSE were measured using ELISA and electrochemical luminescence in 20 children with HFMD complicated by viral encephalitis (encephalitis group), 20 children with HFMD only (simple HFMD group) and 20 healthy children (control group).

RESULTSSerum levels of ferritin in the encephalitis group (212 ± 71 μg/L) were significantly higher than in the simple HFMD group (85 ± 18 μg/L) and control group (70 ± 15 μg/L) (P<0.01). Serum levels of NSE in the encephalitis group (8.6 ± 2.6 μg/L) were also significantly higher than in the simple HFMD group (6.0 ± 1.3 μg/L) and control group (5.6 ± 1.8 μg/L) (P<0.01). Significantly decreased serum ferritin (126 ± 37 μg/L) and NSE levels (6.8 ± 1.9 μg/L) were found in the encephalitis group (P<0.01) after treatment.

CONCLUSIONSSerum levels of ferritin and NSE in children with HFMD complicated by acute viral encephalitis increase, suggesting that serum ferritin and NSE measurement is useful in the early diagnosis of HFMD complicated by acute viral encephalitis.

Acute Disease ; Child, Preschool ; Encephalitis, Viral ; blood ; Female ; Ferritins ; blood ; Hand, Foot and Mouth Disease ; blood ; complications ; diagnosis ; Humans ; Male ; Phosphopyruvate Hydratase ; blood

ObjectiveTo investigate correlation between drug-resistance related genes and mobile genetic elements of Klebsiella pneumoniae resistant to β-lactams. Methods Forty-seven strains of multidrug-resistant Klebsiella pneumoniae were collected from 6 hospitals in Hangzhou and Huzhou of Zhejiang province from August 2008 to May 2010.Modified Hodge test was performed to detect phenotypes of carbapenemases.Forty kinds of β-lactamases (class A-D),ompK35,ompK36,and 12 kinds of mobile genetic elements were detected by PCR,and the results were analyzed by index cluster.ResultsThirty-five strains were positive in modified Hodge test,and 5 kinds of β-lactamases gene ( including KPC-2-like,GenBank:HQ258934) and 9 kinds of mobile genetic elements were detected.Mutations were observed in ompK35 and ompK36 when compared with sensitive strains.Index cluster analysis showed that correlation existed between KPC-2,KPC-2-like and ISKpn6,between TEM-1 and ISEcpl,IS26,int Ⅰ 1,trbC,IS903,and between CMY-2,OXA-30,DHA-1 and tnpU,tnp513,trbC.ConclusionsFive kinds of β-1actamases genes,and mutations in ompK35 and ompK36 may be associated with the resistance to β-1actams in multidrug-resistant Klebsiella pneumoniae.

ObjectiveTo evaluate the prognostic value of MTV on 18F-FDG PET/CT in patients with esophageal cancer.MethodsForty-nine patients with esophageal cancer underwent 18 F-FDG PET/CT scan before surgery.The median follow-up time for the patients was 29 months (range,8- 57 months).The prognostic significance of MTV,age,sex,histologic grade,SUVmax of the primary tumor,tumor size measured on PET/CT,T stage,N stage,M stage,American Joint Committee on Cancer (AJCC) stage,number and location of lymph nodes metastases were assessed by Kaplan-Meier analysis and multivariate Cox model.ResultsIn the univariate analysis,AJCC stage (x2 =16.206,hazard ratio (HR) =1.177,P ＜0.001),N stage (x2 =9.536,HR =10.833,P =0.002),T stage (x2 =5.810,HR=2.397,P=0.016),number of lymph nodes metastases (x2 =11.423,HR =1.567,P =0.001 ),and MTV (x2 =3.872,HR =2.433,P =0.049 ) were significant predictors of survival.Multivariate analysis showed that MTV and AJCC stage were independent predictors of survival (x2 =4.525,HR1.170,P =0.033 ;X2=4.875,HR =3.071,P =0.027).Kaplan-Meier survival curves revealed longer survival time of low-MTV group as compared to high-MTV group ( Log-rank,x2 =4.186,P =0.041 ).ConclusionMTV on 18 F-FDG PET/CT may be an independent prognostic factor in patients with esophageal cancer.

OBJECTIVETo evaluate the feasibility of employing metabonomics method in identification of oral pathogenic bacteria.

METHODSThe Streptococcus mutans ATCC25175 and Actinomyces viscosus ATCC15987 were respectively inoculated in same certain culture medium. The growth curves of the inoculated bacteria were drown by turbidimetry. The culture solutions in four different growth phases of the both bacteria were used to test with the 1H-Nuclear magnetic resonance (1H-NMR) spectroscopy respectively. The data of 1H-NMR spectroscopy results were analyzed by principal components analysis (PCA).

RESULTSThe PCA showed the obvious clustering phenomena and the points of two group data stayed differentially together by two clusters. Therefore, the NMR-based metabonomics profiles can discriminate the two different kind of bacteria.

CONCLUSIONThe metabonomics can be expected to be a kind of promising useful method in quick discrimination of oral pathogenic bacteria.

Culture Media ; Magnetic Resonance Spectroscopy ; Metabolomics ; Streptococcus mutans

BACKGROUND AND OBJECTIVEMost patients with esophageal carcinoma have disease in the locally late stage (stage III) when first diagnosed, with surgery as the first treatment of choice. This study analyzed the clinical data of patients with esophageal squamous carcinoma after radical esophagectomy and investigated prognostic factors.

METHODSThe data of 361 patients with esophageal squamous carcinoma who underwent radical esophagectomy and were hospitalized at Sun Yat-sen University Cancer Center between January 1997 and March 2004 were analyzed. The Kaplan-Meier method was used to analyze prognosis, log-rank test was used to compare the groups, and the Cox proportional hazards model was used for multivariate analysis.

RESULTSThe 1-, 2-, 3-, 4-, and 5-year survival rates were 67.7%, 40.6%, 27.5%, 23.4%, and 20.1%, respectively. Based on univariate analysis, the degree of invasion, rate of lymph node metastasis, number of metastatic regions, number of metastatic lymph nodes, postoperative complications, and duration of surgery were prognostic factors. Based on multivariate analysis, the degree of invasion, rate of lymph node metastasis, and postoperative complications were independent factors for the prognosis.

CONCLUSIONSOf all clinical and pathologic factors, the degree of invasion, rate of lymph node metastasis, and postoperative complications were independent prognostic factors for the patients with stage-III esophageal squamous carcinoma after radical esophagectomy.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; pathology ; surgery ; Esophageal Neoplasms ; pathology ; surgery ; Esophagectomy ; methods ; Female ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Postoperative Complications ; Proportional Hazards Models ; Retrospective Studies ; Survival Rate

OBJECTIVETo evaluate the role of endoscopic ultrasonography (EUS) and CT in the prediction of the resectability of esophageal carcinoma.

METHODSA retrospective study was carried out in 746 patients with esophageal carcinoma. These patients were divided into CT group (480 cases), EUS group (151 cases) and EUS+CT group (115 cases). Images of EUS and CT were double-blindly reviewed by radiologists. Relationship of EUS and CT images with surgical and pathological findings was examined.

RESULTSResection rates in the EUS group, CT group and EUS+CT group were 93.4%, 91.0% and 93.9%, respectively (chi(2)=1.551, P=0.484). Accuracy, sensitivity, specificity, positive predictive value and negative predictive value in the CT group were 81.7%, 87.4%, 23.3%, 92.0% and 15.4%, respectively; 94.7%, 98.6%, 40.0%, 95.9% and 66.7% in the EUS group; and 96.5%, 99.1%, 57.1%, 97.3% and 80.0% in the EUS+CT group, respectively. When assessing aortic invasion, accuracy, sensitivity, specificity, positive predictive value and negative predictive value were 91.3%, 33.3%, 93.1%, 13.5% and 97.7%, in the CT group, respectively; 98.7%, 87.5%, 99.3%, 87.5% and 99.3% in the EUS group, respectively,and 98.3%, 85.7%, 99.1%, 85.7% and 99.1% in the EUS+CT group, respectively. In assessing tracheobronchial invasion, accuracy, sensitivity, specificity, positive predictive value and negative predictive value were 91.3%, 20.8%, 95.0%, 17.9% and 95.8% in the CT group, respectively; 96.0%, 20.0%, 98.6%, 33.3% and 97.3% in the EUS group, respectively; and 98.3%, 66.7%, 99.1%, 66.7% and 99.1% in the EUS+CT group. Differences in assessing resectability were significant between CT group and EUS group (chi(2)=15.131, P=0.000), between CT group and EUS+CT group (chi(2)=15.662, P=0.000), and between EUS group and EUS+CT group (chi(2)=0.502, P=0.346). Differences in assessing aortic invasion were significant between CT group and EUS group (chi(2)=9.764, P=0.000), and between CT group and EUS+CT group (chi(2)=6.659, P=0.004), but were not significant between EUS group and EUS+CT group (chi(2)=0.076, P=0.581). Differences in assessing tracheobronchial invasion were significant between CT group and EUS+CT group (chi(2)=6.659, P=0.004), but were not significant between CT group and EUS group (chi(2)=3.729, P=0.034) and between EUS group and EUS+CT group (chi(2)=1.117, P=0.248).

CONCLUSIONSEUS is a better procedure than CT in the prediction of the resectability and aortic invasion in esophageal carcinoma. There is limited value for EUS and CT in assessing tracheobronchial invasion. Combination of CT and EUS does not improve the prediction of resectability significantly.

Adult ; Aged ; Endosonography ; Esophageal Neoplasms ; diagnostic imaging ; surgery ; Esophagectomy ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tomography, X-Ray Computed ; Treatment Outcome

BACKGROUNDPrevious studies have shown that resveratrol increases endothelial progenitor cell (EPC) numbers and functional activity. Increased EPC numbers and activity are associated with the inhibition of EPC senescence. In this study, we investigated the effect of resveratrol on the senescence of EPCs, leading to potentiation of cellular function.

METHODSEPCs were isolated from human peripheral blood and identified immunocytochemically. EPCs were incubated with resveratrol (1, 10, and 50 µmol/L) or control for specified times. After in vitro cultivation, acidic β-galactosidase staining revealed the extent of senescence in the cells. To gain further insight into the underlying mechanism of the effect of resveratrol, we measured telomerase activity using a polymerase chain reaction (PCR)-enzyme-linked immunosorbent assay (ELISA) technique. Furthermore, we measured the expression of human telomerase reverse transcriptase (hTERT) and the phosphorylation of Akt by immunoblotting.

RESULTSResveratrol dose-dependently inhibited the onset of EPC senescence in culture. Resveratrol also significantly increased telomerase activity. Interestingly, quantitative real-time PCR analysis demonstrated that resveratrol dose-dependently increased the expression of the catalytic subunit, hTERT, an effect that was significantly inhibited by pharmacological phosphatidylinositol 3-kinase (PI3-K) blockers (wortmannin). The expression of hTERT is regulated by the PI3-K/Akt pathway; therefore, we examined the effect of resveratrol on Akt activity in EPCs. Immunoblotting analysis revealed that resveratrol led to dose-dependent phosphorylation and activation of Akt in EPCs.

CONCLUSIONResveratrol delayed EPCs senescence in vitro, which may be dependent on telomerase activation.

Cells, Cultured ; Cellular Senescence ; drug effects ; Endothelial Cells ; cytology ; drug effects ; enzymology ; Humans ; Stem Cells ; cytology ; drug effects ; enzymology ; Stilbenes ; toxicity ; Telomerase ; metabolism

Objective To investigate the resistant mechanism of quinolones on multi-drug resistant Klebsiella caused pneumonia(MDR-KPN).Methods From August 2008 to May 2010,47 strains of MDR-KPN were collected from 6 hospitals in Hangzhou and Huzhou in Zhejiang province in China.Drug target genes to quinolones (gyrA,parC) and quinolone-resistance genes mediated by mobile genetic elements [qnrA,qnrB,qnrS,aac (6')-Ⅰ b-cr,qepA] were analyzed by PCR and verified by DNA sequencing.Results Positive results were found in 47 strains of MDR-KPN,43 strains (91.5％) of gyrA mutation,40 strains(85.1％) ofparC mutation,3 strains (6.4％) of qnrB2,1 strain (2.1％) ofqnrB 4,8 strains (17.0％) ofqnrS 1,5 strains (10.6％) of qnrS 4,2 strains (4.3％)of aac (6')-Ⅰ b-cr respectively.Moreover,5 novel variants of gyrA (GenBank accession number:JN811952,JN811953,JN811954,JN811955,JN811956),5 novel variants of parC (GenBank accession number:JN817432,JN817433,JN817434,JN817435,JN817436)were also identified.In addition,qnrS4 (GenBank accession number:JN836269) appeared to be the novel variants of qnrS.Conclusion Quinolone-resistance-determining region played a key role on the resistance to quinolones in this group of MDR-KPN,and quinolone-resistance genes mediated by mobile genetic elements [qnrB2,qnrB4,qnrS1,qnrS4,aac (6')-Ⅰ b-cr] showed positive in some parts of the strains.This was the first report on emergence of qnrS4 in the world.