Objective To investigate the factors affecting the isolation of fastidious bacteria such as Streptococcus pneumoniae and Haemophilus influenzae from lower respiratory tract specimens in children.Methods A total of 210 lower respiratory tract specimens from children were collected and inoculated on both blood agar plate and chocolate agar plate 0.5 h,2 h,6 h,12 h,and 24 h after collection.The effect of specimen turnaround time,transport medium,and inoculation medium on the isolation of fastidious bacteria was studied.Results The results of 200 qualified specimens showed that the isolation of S.pneumoniae and H.influenzae significantly decreased with the increasing of specimen turnaround time (P＜0.05),but no significant difference was observed between 0.5 h and 2 h timepoints.The isolation of fastidious bacteria in semi-solid medium was non-significantly higher than the other two transport medium (P＞0.05).No significant difference was found in the isolation and growth ofS.pneumoniae and H.influenzae on different inoculation media (P＞0.05).Conclusions The appropriate specimen turnaround time,transport medium,and inoculation medium are important for improving the isolation of fastidious bacteria from lower respiratory tract in children.

Objective To investigate the distribution and resistance profile of bacterial isolates in Shanghai Children's Hospital. Methods Antimicrobial susceptibility of all isolates was determined by Kirby-Bauer disk diffusion method according to 2016 CLSI standard. The data were analyzed by WHONET 5.6 software. Results A total of 23 259 non-duplicate strains were isolated from 2011 to 2016, including 10 885(46.8％)gram-postive cocci and 12 374(53.2％)gram-negative bacilli. The average prevalence of methicillin-resistant S. aureus(MRSA)and coagulase-negative Staphylococcus(MRCNS)was 35.8％ and 82.2％, respectively. The prevalence of MRSA rose from 27.4％ in 2011 to 42.9％ in 2016. The resistance rate of MRSA and MRCNS strains were significantly higher than methicillin sensitive strains. The resistance rate of Enterococcus faecium strains was significant higher than Enterococcus faecalis. The prevalence of non-susceptible Streptococcus pneumoniae was 31.2％(908). No gram-positive strain was resistant to vancomycin or linezolid. The prevalence of carbapenem resistance increased in gram-negative strains. The resistance rate of Klebsiella pneumoniae to imipenem and meropenem rose from 3.1％ and 4.8 ％ in 2011 to 28.7％ and 37.4％ in 2016, respectively.The rate of Pseudomonas aeruginosa resistance to imipenem and meropenem rose from 13.8％ and 16.5％ in 2011 to 18.8％ and 19.4％ in 2016, respectively, while Acinetobacter baumannii showed resistance rate of 38.3％ and 39.9 ％ in 2011 to 68.4％ and 69.7％ in 2016. Conclusions Increasing prevalence of MRSA, carbapenem-resistant K. pneumoniae, extensively drug-resistant A.baumannii has become a concern in clinical practice. Therefore, antimicrobial resistance surveillance should be highly strengthened in children's hospital.

Objective To investigate the effect of tigecycline on multi-drug resistant Acinetobacter baumannii ( MDR -AB ) adeB gene expre-ssion and its clinical significance.Methods A total of 60 clinical iso-lates of multidrug-resistant Acinetobacter baumannii were selected, from which 15 experimental strains with positive result to ciprofloxacin ( CIP) efflux pump encoded gene were screened.The minimum inhibitory con-centration ( MIC) before and after given tigecycline was determined using agar dilution assay, the gene of the strains with large change of MIC value was sequenced and compared.Results The lowest MIC of tigecy-cline to MDR-AB was 2 mg? mL-1 .The MDR-AB′s MIC of tigecy-cline at concentration of 0 , 0.62 mg? mL-1 and ciprofloxacin at 1.25 , 2.5 mg? mL-1 were significantly different ( P<0.05).After tigecycline treatment, adeB gene segment had 15 bases mutation, 5 amino acids are replaced and 4 mutant amino acid had no substitution.The adeB gene expression levels before and after the treatment of tigecyclin were (2.51 ±0.72) and (0.87 ±0.21), statistically significant (P<0.05). Conclusion Tigecycline inhibits MDR -AB efflux pump adeB gene expression, lowers adeB expression levels. Other agents with its combination for the treatment of MDR-AB might improve the clinical efficacy.

Objective:To explore the role and mechanism of Forkhead box protein 3 (FoxP3)in biological behavior of gastric cancer (GC)cells.Methods:Invitro,the plasmid of FoxP3-shRNA was transfected into GC cells,and then the stably transfected cells were established by drug-screening and monoclone-selection.MTT assay was used to detect the growth and proliferation of GC cells.And cell cycle was detected by flow cytometry.MTT assay was used to measure the difference regarding sensitivity to chemotherapy drugs.Results:GC cell line which was stably transfected with FoxP3 gene,was established.GC cell line with up-regulation of FoxP3 gene,compared to the vector-transfected control,showed slower growth and proliferation rate,weaker ability of invasion (transmembrane cell counts:[203±42]cells/HPvs [891±100]cells/HP, P<0.05),and higher sensitivity to chemotherapy drugs(P<0.05).Conclusions:FoxP3 gene plays a role in inhibiting the growth of GC cells.

OBJECTIVEMonitoring of minimal residual disease (MRD) is proven to be increasingly valuable for predicting relapse and outcome of childhood acute lymphoblastic leukemia (ALL) and is used to identify patients' risk groups in several current clinical trials. However, the limitation is that most studies focused on the cut-off value at 10(-4) and the time point after induction. The aim of this study was to investigate the predictive values of different MRD levels detected at different chemotherapy phases in childhood ALL.

METHODSOne hundred and two patients were enrolled in this study from January 2002 to December 2004 in our hospital. All the patients were treated with modified National Protocol of Childhood ALL in China 1997. MRD levels were detected on the 15th day, 29th day, at 3 months, 6 months and 12 months after initial chemotherapy. All samples were stained with a panel of four colour combinations of fluorochrome conjugated monoclonal antibodies according to the leukemia-associated immunophenotype (LAIP) defined at diagnosis and analyzed by multi-parametric flow cytometry. CD45CD19CD34CD10, CD45CD19CD34CD20 and CD45CD19CD10CD20 were the most common combinations in B lineage ALL, while CD45CD2CD3CD7 and CD45CD2CD3CD34 were the most frequently used immunophenotypes for T lineage ALL. The median follow-up time was 63.3 months ranged from 40.6 to 87.5 months.

RESULTSOf the 102 patients, 64 were male and 38 were female, with a median age of 5.7 (0.2 - 14.8) years. Eighty-eight cases were diagnosed as B lineage ALL and the remaining 14 were T-ALL. The 5-year overall survival (OS) rate and event free survival (EFS) rate for this cohort were (86.9 +/- 3.4)% and (79.9 +/- 4.0)%, respectively. Twelve patients underwent relapse. Among the 102 patients, 14.3% had negative MRD (MRD < 10(-4)) on day 15, 43.9% on day 29, 39.1%, 39.7% and 45.6% had negative MRD at the third, sixth and twelfth month after chemotherapy. Patients who could achieve negative MRD within one year had superior outcome to the others [5-year EFS rates: (92.5 +/- 3.2)% vs. (58.3 +/- 8.6)%, P < 0.001]. The EFS for patients based on MRD levels measured at different stages of therapy were compared by Kaplan-Meier analyses. MRD was predictive of outcome at all 5 time points at a range of thresholds. The optimum threshold, selected for each time point on the basis of log rank analysis, progressively dropped from 10(-2) of day 15 [5-year EFS rates (79.8 +/- 10.3)% vs. (28.6 +/- 17.1)%, P < 0.001], to 10(-3) of day 29 [5-year EFS rates (88.3 +/- 4.9)% vs. (51.3 +/- 14.4)%, P < 0.003], to 10(-4) at 3 [5-year EFS rates (92.4 +/- 5.1)% vs. (65.5 +/- 7.5)%, P < 0.015], 6 [5-year EFS rates (96.3 +/- 3.6)% vs. (65.4 +/- 7.5)%, P < 0.003] and 12 [5-year EFS rates (100.0 +/- 0.0)% vs. (67.7 +/- 8.4)%, P < 0.002] months. And the hazard ratios for relapse and death at higher MRD level groups were 5.91 (95%CI: 1.9 - 18.9), 5.02 (95%CI: 1.5 - 16.5), 5.21 (95%CI: 1.2 - 22.9) and 11.10 (95%CI: 1.5 - 84.5) on day 15, day 29, at month 3 and month 6, respectively. And MRD >or= 10(-2) on day 15 was proven to be an independent predictor by multivariate Cox proportional-hazards regression model.

CONCLUSIONDynamic MRD detection by multi-parametric flow cytometry is highly predictive of outcome for childhood ALL, and the cut-off values at different time points were different.

Adolescent ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Infant ; Infant, Newborn ; Male ; Neoplasm, Residual ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; pathology ; Predictive Value of Tests ; Prognosis

OBJECTIVEThe patients with recurrent or refractory neuroblastoma have a very poor prognosis and high mortality. 10-hydroxycamptothecin (HCPT) is a new agent extracted from comptotheca acuminate, a native plant. It has been shown to be very effective in some solid tumors such as gastric and colon cancers, lung cancers and ovary cancers. However, its efficacy in neuroblastoma has not been determined. This study aimed to investigate the therapeutic effects of HCPT in the treatment of recurrent or refractory neuroblastoma in children.

METHODSTen children with recurrent neuroblastoma and two children with refractory neuroblastoma were treated with HCPT. Of them, 5 children with recurrent neuroblastoma were treated with HCPT alone, and the other 7 patients received combination chemotherapy of HCPT plus other agents. The HCPT alone treatment group was injected with HCPT (7.5 mg/m2 daily) for 14 consecutive days. The combination chemotherapy group was alternately treated with the modified new protocol A1 (cyclophosomide 1 200 mg/m2 on day 1, etoposide 100 mg/m2 on days 1-5, HCPT 5 mg/m2 on days 1-3, cisplatin 90 mg/m2 on day 4) and the modified protocol B (ifosfomide 1.5 g/m2 on days 1-5, HCPT 5 mg/m2 on days 1-3, carboplatin 450 mg/m2 on day 2).

RESULTSFour patients (33.3%) achieved partial remission and 8 patients (66.7%) had stable disease. The median remission duration was 3.5 months (2-5 months). HCPT treatment as a single agent resulted in mild side effects. Myelosuppression and digestive disorders were found as the main adverse events in the combined chemotherapy group. No chemotherapy related deaths were found.

CONCLUSIONSHCPT is safe and effective in the treatment of recurrent or refractory neuroblastoma. The toxicities of HCPT are tolerable. The long-term efficacy of HCPT warrants further research.

Adolescent ; Antineoplastic Agents, Phytogenic ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Camptothecin ; analogs & derivatives ; therapeutic use ; Child ; Child, Preschool ; Female ; Humans ; Male ; Neuroblastoma ; drug therapy ; Recurrence

OBJECTIVETo provide reliable evidence of "J in three-needle therapy" for treatment of stroke.

METHODSMulti-central randomized controlled trials were adopted, 180 hemiplegia patients of ischemic stroke were randomly divided into a fin three-needle group (90 cases) and a routine acupuncture group (90 cases). Two groups were both treated with basic neurology therapies, and J in three-needle group was treated with J in three-needle therapy, three acupoints of tempora, hand and foot etc. were selected; the routine acupuncture group was treated with traditional acupuncture, Quchi (LI 11), Huantiao (GB 30), Futu (ST 32) etc. were selected. Both groups were treated with acupuncture for 5 weeks. The cognitive function score of functional comprehensive assessment scale (FCA), the scores of mini-mental state examination scale (MMSE) and modified Barthel index (BI) were compared before and after treatment between two groups. Results After treatment, the scores of FCA, MMSE and BI in both groups were significantly improved compared to those before treatment (P < 0.01, P < 0.05); the improvement of FCA score, MMSE score and BI score in the J in three-needle group were superior to those of the routine acupuncture group after treatment (P < 0.01, P < 0.05). The total effective rate of 85.4% in the J in three-needle group was superior to tohat of 70.0% in the routine acupuncture group (P < 0.05).

CONCLUSIONJ in three-needle acupuncture treatment can obviously improve the cognitive function and activity ability of daily life of hemiplegia patients after stroke, and the therapeutic effect of J in three-needle therapy is superior to that of traditional acupuncture treatment.

Activities of Daily Living ; Acupuncture Therapy ; Adult ; Aged ; Cognition ; Female ; Hemiplegia ; etiology ; psychology ; rehabilitation ; therapy ; Humans ; Male ; Middle Aged ; Stroke ; complications

Objective:To investigate the tendency of bacterial distribution and drug resistance of clinically isolated pathogens in the pediatric intensive care unit (PICU), which provided references for the reasonable application of antibiotics.Methods:The distribution characteristics of all clinical isolates from PICU of Children′s Hospital of Shanghai Jiaotong University from January 2010 to December 2018 and their trend of drug resistance were retrospectively analyzed.Results:A total of 2 749 strains of bacteria were isolated, including 1 912 strains (69.6%) Gram-negative bacteria and 837 strains (30.4%) Gram-positive bacteria.The top 6 detected bacteria were Acinetobacter baumannii (749 stains, 27.2%), Klebsiella pneumoniae (289 stains, 10.5%), Staphylococcus aureus (214 stains, 7.8%), Stenotrophomonas maltophilia (207 stains, 7.5%), Escherichia coli (204 stains, 7.4%) and Pseudomonas aeruginosa (189 stains, 6.9%). Among them, the detective rate of Maltophilia Stenotrophomonasannually increased from 6 strains (2.8%) in 2010 to 39 strains (9.5%) in 2018.The resistance rates of Acinetobacter baumannii and Klebsiella pneumoniae to carbapenems increased year by year, which was up to 96.0% and 71.4% to Meropenem by 2018.Their resistance rates to the third-generation cephalosporins, aminoglycosides and sulfonamides were higher than 70.0%.The sensitivity rate to Tigecycline and Polymyxin was 100.0%.The detection rate of Methicillin-resistant Staphylococcus aureus (MRSA) significantly increased from 18.2% in 2010 to 50.0% in 2018 ( χ2=19.38, P=0.013). No Vancomycin-resistant strains were found. Conclusions:Gram-negative bacteria are the main clinical isolates of PICU.Acinetobacter baumannii, Klebsiella pneumoniae, and especially Pseudomonas maltophilus, have a significant growth trend in the detection rate. Acinetobacter baumannii and Klebsiella pneumoniae are highly resistant to carbapenems.MRSA annually grows, but it still maintains a high degree of sensitivity to Vancomycin.

Objective: To observe the blood perfusion volume variation pattern in the body surface microcirculation at the Yuan-Primary and the Xi-Cleft points during the menstrual cycle in female college students with moderate constitution under normal physiological state of the uterus; to explore the specific laws of the body surface microcirculation at the Yuan-Primary and Xi-Cleft points in response to the uterine qi and blood changes under normal physiological conditions, and to provide the experimental basis for the specificity of acupoints reflecting the uterine function. Methods: Forty-three healthy and moderate constitution female college students with regular menstrual cycles, without dysmenorrhea and not yet giving birth were recruited. Bilateral Yuan-Primary points [Taichong (LR 3), Taibai (SP 3) and Taixi (KI 3)] and Xi-Cleft points [Zhongdu (LR 6), Diji (SP 8) and Shuiquan (KI 5)], belonging to the three yin meridians of foot and adjacent to the spinal cord segment of the uterus, were selected as the detection acupoints; the crossing point of the three yin meridians of foot [Sanyinjiao (SP 6)], the uterus-related meridian acupoint [Xuehai (SP 10)], the uterus-non-related meridian acupoint [Xuanzhong (GB 39)], and the non-meridian non-acupoint point were selected as the control points. The laser speckle blood flow imaging technique was used to monitor the blood perfusion volume in skin microcirculation at the above points at the menstrual, follicular, ovulatory, and luteal phases of the subjects. Results: The blood perfusion volume in the body surface microcirculation at the right Zhongdu (LR 6) at the ovulatory phase was higher than that at the menstrual, follicular and luteal phases (all P<0.05); there was no significant difference in the microcirculation blood perfusion volume at the other points among different phases (all P>0.05). Conclusion: The blood perfusion volume in the body surface microcirculation at Zhongdu (LR 6), the Xi-Cleft point of the Liver Meridian, shows a specific response to qi and blood changes in the uterus of women with moderate constitution.

The sialyl Lewis X (SLex) antigen encoded by the FUT7 gene is the ligand of endotheliam-selectin (E-selectin).The combination of SLex antigen and E-selectin represents an important way for malignant tumor metastasis.In the present study,the effect of the SLeX-binding DNA aptamer on the adhesion and metastasis of hepatocellular carcinoma HepG2 cells in vitro was investigated.Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence staining were conducted to detect the expression of FUT7 at both transcriptional and translational levels.The SLex expression in HepG2 cells treated with different concentrations of SLeX-binding DNA aptamer was detected by flow cytometry.Besides,the adhesion,migration,and invasion of HepG2 cells were measured by cell adhesion assay,and the Transwell migration and invasion assay.The results showed that the FUT7 expression was up-regulated at both mRNA and protein levels in HepG2 cells.SLeX-binding DNA aptamer could significantly decrease the expression of SLex in HepG2 cells.The cell adhesion assay revealed that the SLeX-binding DNA aptamer could effectively inhibit the interactions between E-selectin and SLex in the HepG2 cells.Additionally,SLeX-binding DNA aptamers at 20 nmol/L were found to have the similar effect to the monoclonal antibody CSLEX-1.The Transwell migration and invasion assay revealed that the number of penetrating cells on the down-side of Transwell membrane was significantly less in cells treated with 5,10,20 nmol/L SLeX-binding DNA aptamer than those in the negative control group (P＜0.01).Our study demonstrated that the SLeX-binding DNA aptamer could significantly inhibit the in vitro adhesion,migration,and invasion of HepG2 cells,suggesting that the SLeX-binding DNA aptamer may be used as a potential molecular targeted drug against metastatic hepatocellular carcinoma.