Objective To investigate the distribution and evolvement of resistance profiles of the pathogens isolated from children diagnosed with urinary tract infection (UTI).Methods The pediatric patients who were diagnosed with UTI in Children's Hospital of Fudan University from January 2001 to December 2010 were enrolled in this retrospective study.The clean catch midstream urine samples were collected and the pathogens were isolated by culture.Bacterial susceptibility test was performed by following a protocol of the standardized Kirby Bauer (KB) method.The enumeration data were analyzed by chi square test and the tendency of drug resistant rate was analyzed by curvilinear regression.Results A total of 10 686 pathogen strains were isolated during the past 10years.The major pathogen was Escherichia coli which counted for 38％ -56％ of the isolates.However,the proportion of Enterococcus faecalis (0 - 17％,F =34.075,P =0.000) and Enterococcus faecium (3 ％ - 15 ％,F =7.139,P =0.028) increased rapidly.The resistant rates of Escherichia coli to cefazolin,cefuroxime sodium,cefotaxime and ceftazidime were 35.8％- 62.3％(F=215.735,P=0.000),33.1％ -57.0％(F=70.674,P=0.000),19.0％ -56.5％(F=52.355,P=0.000) and 2.1％-23.5％(F=16.807,P=0.003),respectively.The resistant rate of Escherichia coli to ciprofloxacin was 23.4％ - 33.8 ％ (F=0.989,P=0.349),while the resistant rate of Escherichia coli to amikacin and cefoperozone/sulbactam were both less than 7％.There was no Escherichia coli isolate showing resistant to imipenem.The resistant rates of Enterococcus faecium to ampicillin and ciprofloxacin were higher than 60.0％ and 50.0％,respectively,while the resistant rate of Enterococcus faecalis to these two antibiotics decreased from 40.0％ to 9.6 ％ (F =17.497,P =0.009) and 60.0％ to 21.6％ (F=12.826,P=0.009),respectively.The resistant rate of Enterococcus faecium to nitrofurantoin decreased from 32 ％ (in the year of 2002) to 9.4％ (in the year of 2010,F=34.075,P=0.000) and the resistant rate of Enterococcus faecalis decreased from 9.2％ (in the year of 2005) to 1.4％ (in the year of 2010,F=7.139,P=0.028).The resistant rates to erythromycin were both higher than 75.0％ in both Enterococcus in recent 10 years.There was no Enterococcus isolates showing resistant to vancomycin.Conclusions Escherichia coli,Enterococcus faecalis and Enterococcus faecium are the most common pathogens associated with UTI in children.The proportions of Enterococcus faecalis and Enterococcus faecium among the pathogens of UTI are increasing.The antimicrobial activity of ampicillin to Enterococcus faecalis keeps at a high level.However,there are few drugs can be used for treating Enterococcus faecium infection.

Objective To investigate the etiological and epidemiological characteristics of fungal keratitis in Jilin province of China and to establish a rapid and specific method for molecular identification of the prevalent fungal pathogens.Methods Corneal scrapings were collected from 225 patients with suspected fungal keratitis.Fungal strains were isolated and identified based on their morphology and physiological characteristics.The epidemiological characteristics of all isolated strains causing fungal keratitis were statistically analyzed.Species-specific primers of Fusarium solani (F.solani) were designed and used together with the universal fungal primers to establish a multiplex PCR assay for identification of F.solani in corneal scrapings.Results 156 out of 225 patients (69.3％) were diagnosed as fungal keratitis by fungal culture followed by the examination of morphological and physiological characteristics.A total of 168 pathogenic fungi strains were isolated,most of which were Fasarium spp.(49.4％),followed by Aspergillus spp.(17.9％) and Candida spp.(14.3％).F.solani was the predominant pathogen accounting for 34.5％ in all patients.Most of the patients (87.5％) were farmer and male patients (57.1％) accounted for the majority of 156 patients as well.Corneal trauma (38.5％) was considered as the main predisposing factor.The established multiplex PCR could specifically amplify a 300 bp nucleotide fragment of F.solani.It could be used for a rapid identification of F.solani in corneal scrapings.Conclusion Fusarium genus,particularly the species of F.solani,was the predominant pathogen for fungal keratitis in Jilin province of China.Corneal trauma was the most important predisposing factor.The established multiplex PCR could identify fungal infection from corneal scrapings rapidly and specifically.These findings are very important for the early diagnosis and treatment of fungal keratitis.

Objective To observe the expression of cyclooxygenase(COX)-2 in different malignancy astrocytoma by immunohistochemistry,so as to judge the invasive action of COX-2 in astrocytoma.Methods A total of 55 cases with astrocytoma underwent 63 times operation including 8 recurrent cases were selected,and 9 cases with severe brain injury who needed decompression surgery were selected as control group.The astrocytoma tissues and decompression surgery to remove tissues removed immediately fixed in 10％ formalin buffer solution.The expression of COX-2 was determined by immunohistochemistry.Results The positive expression rate of COX-2 in astrocytoma tissues was higher than that in decompression surgery to remove tissues[69.84％ (44/63) vs.1/9],and there was significant difference (x2 =11.589,P ＜ 0.01).The positive expression rate of COX-2 in recurrent astrocytoma tissues was higher than that in primmy astrocytoma tissues [8/8 vs.65.45％ (36/55)],and there was significant difference (x2 =3.957,P ＜ 0.05).With increased astrocytoma malignancy,the positive expression rate of COX-2 also increased,and there was significant difference (P ＜ 0.05).Conclusion COX-2 may have some relationship with the invasion of astrocytoma,leading to increase tumor grade,and relapse.

Since the initial observations of oral bacteria within dental plaque by van Leeuwenhoek using his primitive microscopes in 1680, an event that is generally recognized as the advent of oral microbiological investigation, oral microbiology has gone through phases of "reductionism" and "holism". From the small beginnings of the Miller and Black period, in which microbiologists followed Koch's postulates, took the reductionist approach to try to study the complex oral microbial community by analyzing individual species; to the modern era when oral researchers embrace "holism" or "system thinking", adopt new concepts such as interspecies interaction, microbial community, biofilms, poly-microbial diseases, oral microbiological knowledge has burgeoned and our ability to identify the resident organisms in dental plaque and decipher the interactions between key components has rapidly increased, such knowledge has greatly changed our view of the oral microbial flora, provided invaluable insight into the etiology of dental and periodontal diseases, opened the door to new approaches and techniques for developing new therapeutic and preventive tools for combating oral polymicrobial diseases.
Bacteria ; classification ; Bacterial Infections ; prevention & control ; Bacterial Physiological Phenomena ; Biofilms ; Dental Plaque ; microbiology ; Humans ; Mouth ; microbiology ; Periodontal Diseases ; microbiology ; prevention & control ; Tooth Diseases ; microbiology ; prevention & control

Objective To evaluate the influence of narrow-band ultraviolet B on lesional microvessel density (MVD),vascular endothelial growth factor (VEGF),matrix metalloproteinases 2 (MMP-2)as well as on serum VEGF in patients with psoriasis vulgaris(PV).Methods Fifteen patients with PV were recruited into this study with 10 normal human controls.All patients received NB-UVB phototherapy thrice a week for 4-5 weeks.Prior and after the treatment,psoriasis area and severity index (PASI)was calculated,tissue specimens were taken from non-photoexposed lesions,and sera samples were obtained from these patients.Then,MvD and the expression level of VEGF and MMP-2 were measured by immunohistochemical labeled dextran polymer(LDP)method in the tissue specimens.Also,the serum level of VEGF was tested by enzyme-linked immunosorbent assay(ELISA).Results PASI score remarkably decreased in patients after the photothempy(t=13.35,P＜0.01).The MVDs were 20.52±5.02,7.33±1.24 and 4.26±0.79 capillaries per high power field(400 × amplification),in psoriatic lesions before treatment,after treatment,and normal control tissues,respectively,with a significant difference among the three groups (F=97.57,P＜0.05),and a significant increase was observed in the lesions before treatment compared with those after treatment and normal controls.The serum level of VEGF was 307.55±121.65 ng/L in psoriatic lesions before treatment,significantly higher than that after treatment(163.92±95.57 ng/L),and in normal control skin (139.78±79.06 ng/L),whereas there was no significant difference between the latter two groups(P＞0.05).The positivity rate of MMP-2 was similar among the three groups without statistical difference(P＞0.05).In psoriatic patients,a positive correlation was observed among PASI score,MVD,lesional and serum VEGF levels(P＜0.05),also among the MVD,VEGF and MMP-2 levels in lesions(P＜0.05).but lesional MMP-2 was unrelated to PASI score or sgrum VEGF(both P＞0.05).Conclusions NB-UVB may regulate superficial dermal microvascular proliferation by acting on the expression of VEGF in sera and lesions of psoriatic patients.VEGF and MMP-2 may bOth participate in the proliferation process of microvessels,while MMP-2 is unlikely to be involved in the therapeutic mechanism of NB-UVB.

Objective To assess the prevalence of Campylobacter in the children with acute bacterial diarrhea in Shanghai.Methods Epidemiological survey.Totally 6 641 children with acute bacterial infectious diarrhea from outpatients and inpatients in Children′s Hospital of Fudan University were submitted to the investigation during January 2011 to December 2012.The Campylobacter was isolated from stool samples collected from subjects in micro aerobic environment and identified by both multi-PCR and matrix-assisted laser desorption ionization-time-of-flight mass spectrometry ( MALDI-TOF MS ) .Antimicrobial susceptibility tests were assayed by disk-diffusion method according to EUCAST standard.The isolates molecular typing was done by PFGE.SPSS16.0 was used to analyze the results.Results A total number of 6 641 subjects were enrolled, among them, 305 patients were infected with Campylobacter.The prevalence rate was 4.6%(305/6 641).Among the infected patients, 240 patients were infected with Campylobacter jejuni and 65 patients were infected with Campylobacter coli, the infectious rates of these two pathogenic bacteria were 3.6%and 1.0%, respectively.The peak infectious rate in patients older than 1 year of age was 6.2%(209/3 385) which was higher than that in children under 1 year of age (2.9%, 96/3 256),χ2 =35.98,P<0.001.The infectious rate in winter and spring (6.8%, 138/2 040) was higher than that in the other seasons ( 3.6%, 167/4 601 ) ,χ2 =28.59, P <0.001.Antimicrobial susceptibility test results showed that 91.5%( 279/305 ) isolates were resistant to ciprofloxacin and 11.8%( 36/305 ) isolates were resistant to erythromycin.A total of 9 genotypes of Campylobacter were found by PFGE cluster analysis.The similarity were ranged from 65.1%-100.0%for type A, 67.6%-100.0%for type B, 61.7%-100.0%for type C, 59.0%-100.0%for type D, 71.4%for type F, 80.0%for type H, 54.4%-90.9%for type I, and only one strain was classified as type E and G.Conclusions Campylobacter is a major pathogenic bacteria associated with acute bacterial infectious diarrhea in children especially in children older than 1 year of age in Shanghai.The prevalent pattern of this pathogen was sporadic and the sharp peak was in winter and spring.The isolates are highly resistant to ciprofloxacin but still sensitive to erythromycin.

AIM:To study the effects of adiponectin on H 2 O2-induced cell injury and tau hyperphosphorylation in human neuroblastoma SH-SY5Y cells.METHODS:Cell viability was determined by MTT assay .H2 O2-induced cell in-jury and morphological changes in the SH-SY5Y cells with or without adiponectin treatment were observed .The level of tau phosphorylation as well as the activities of protein phosphatase 2A (PP2A) and of glycogen synthase kinase-3β(GSK-3β) were examined by Western blotting . RESULTS: Adiponectin significantly attenuated H 2 O2-induced cell injury (P<0.01).Adiponectin upregulated the activity of PP2A and decreased phosphorylation levels of tau under the stimula-tion with H2O2(P<0.01).Okadaic acid, a specific inhibitor of PP2A, blocked the protective effects of adiponectin (P<0.01).Adiponectin increased the phosphorylation level of GSK-3βat Ser9 site under H2O2stimulation (P<0.01).CON-CLUSION:Adiponectin protects SH-SY5Y cells against H2O2-induced cell injury and decreases tau hyperphosphorylation by activating PP2A and inactivating GSK-3β.

BACKGROUND:Bone marrow mesenchymal stem cel s have attracted widespread attention for the capabilities of self-renewal and muti-differentiation, which have been used in treatment of various diseases.
OBJECTIVE:To study the effect of three-dimensional spheroid culture system on the stemness and senescence of bone marrow mesenchymal stem cel s.
METHODS:Mesechyaml stem cel s were isolated from the bone marrow of C57/B6 mice, 3 weeks old, and cultured onto the culture plates coated with or without chitosan. After 5 days of culture, the cel phenotype and expression of stemness related markers CD44 and Sca-1 were analyzed by flow cytometry. PI and Annexin-V staining were used to detect cel apoptosis. Also,β-Gal staining was applied for identification of aging.
RESULTS AND CONCLUSION:The mouse mesenchymal stem cel s began to form spheroids on day 3. The stemness-related markers, including CD44 and Sca-1, expressed higher in spheroid mesenchymal stem cel s than the cel s under normal culturing. Compared with the normal culture group, the apoptosis and senescence of cel s from spheroid culture were lower. The results indicate that the formation of spheroids on chitosan films can increase the stemness, decrease the apoptosis and slow the senescence of mesenchymal stem cel s.

Objective To evaluate the treatment outcome, prognostic factors, radiation dose, and toxicities in patients with early-stage primary diffuse large B-cell lymphoma of Waldeyer's ring (WR-DLBCL) treated with intensity-modulated radiotherapy (IMRT).Methods This study included 80 patients with a confirmed diagnosis of stage Ⅰ-Ⅱ primary WR-DLBCL who were admitted to our hospital from 2008 to 2015.Only 3 patients received radiotherapy alone, and the other patients received radiotherapy and chemotherapy.After chemotherapy, 24 patients achieved complete remission (CR), and 53 patients achieved partial remission (PR).IMRT was given to the primary lesion and cervical lymphatic drainage region.Survival analysis was performed using the Kaplan-Meier method and log-rank test.The Cox model was used for analysis of prognostic factors.The toxicities were scored using the RTOG criteria.Results The median follow-up was 64 months.The 5-year locoregional control (LRC), overall survival (OS), and progression-free survival (PFS) rates were 94%, 88%, and 84%, respectively.The dose-volume histogram showed that the maximum, mean, and minimum doses to primary gross tumor volume were 54.47 Gy, 52.27 Gy, and 38.83 Gy, respectively.Prognostic analysis showed that age>60 years and increased lactate dehydrogenase (LDH) were influencing factors for OS (P=0.009 and 0.002), and that aged>60 years, IPI ≥2, and increased LDH were influencing factors for PFS (P=0.001, 0.035, and 0.007).Among all patients, 12, 53, and 8 experienced grade 1-3 radiation-induced acute oral mucositis, respectively, and 16 and 13 experienced grade 1 and 2 xerostomia as the late toxicity, respectively.Conclusions For patients with early-stage primary WR-DLBCL, IMRT results in satisfactory OS, PFS, and LRC and has tolerable early or late radiation-induced toxicities.

The study investigated the effects of heat shock protein 70 (HSP70) antisense oligonucleotide (ASODN) on the proliferation and apoptosis of a human hepatocellular carcinoma cell line (SMMC-7721 cells) in vitro. HSP70 oligonucleotide was transfected into SMMC-7721 cells by the mediation of Sofast transfection reagent. Inhibition rate of SMMC-7721 cells was determined by using MTT method. Apoptosis rate and cell cycle distribution were measured by flow cytometry. Immunocytochemistry staining was used to observe the expression of HSP70, Bcl-2 and Bax. The results showed that HSP70 ASODN at various concentrations could significantly inhibit the growth of SMMC-7721 cells, and the inhibition effect peaked 48 h after transfection with 400-nmol/L HSP70 ASODN. Cytometric analysis showed the apoptotic rate was increased in a dose- and time-dependent manner in the HSP70 ASODN-treated cells. The percentage of cells in the G(2)/M and S phases was significantly decreased and that in the G(0)/G(1) phase increased as the HSP70 ASODN concentration was elevated and the exposure time prolonged. Immunocytochemistry showed that treatment of SMMC-7721 cells with HSP70 ASODN resulted in decreased expressions of HSP70 and Bcl-2 proteins, and an increased expression of Bax protein. It was concluded that the HSP70 ASODN can inhibit the growth of the SMMC-7721 cells and increase cell apoptosis by down-regulating the expression of HSP70. HSP70 ASODN holds promise for the treatment of hepatocellular carcinoma.