Objective: To compare the efficacy of induction chemotherapy (IC) plus intensity-modulated radiotherapy (IMRT) with that of concurrent chemo-radiotherapy (CCRT) plus adjuvant chemotherapy (AC) for patients with loco-regionally advanced naso-pharyngeal carcinoma (NPC). Methods:Data of 240 patients with loco-regionally advanced NPC were reviewed. These patients were admitted to the Sun Yat-sen University Cancer Center between January 2004 and December 2008. Among the 240 patients, 117 under-went the IC+IMRT and 123 were treated with the CCRT+AC. The IC+IMRT group received a regimen including cisplatin and 5-fluoro-uracil (5-FU). The CCRT+AC group received cisplatin concurrently with radiotherapy and subsequently received adjuvant cisplatin and 5-FU. The survival rates of the patients were assessed by Kaplan-Meier analysis, and the survival curves were compared by Log-rank test. Multivariate analysis was conducted using Cox proportional hazard regression model. Results:The 5-year overall survival (OS), disease-free survival, distant metastasis-free survival, local relapse-free survival, and the nodal relapse-free survival were 78.0%versus 78.7%, 68.9%versus 67.5%, 79.0%versus 77.0%, 91.6%versus 91.0%, and 95.3%versus 93.7%in the IC+IMRT and CCRT+AC groups, respectively. The survival between the two groups exhibited no significant differences. Higher rates of Grades 3 to 4 nau-sea-vomiting (8.1%vs. 1.7%, P=0.023) and leukopenia (9.7%vs. 0.9%, P=0.006) were observed in the CCRT+AC group. Multivariate analysis revealed that N stage and age were significant prognostic factors for the OS of the patients with loco-regionally advanced NPC. Conclusion:The treatment outcomes of IC+IMRT and CCRT+AC were similar. Distant metastasis remained as the predominant mode of treatment failure.

Objective To evaluate the treatment effect of bladder instillation chemotherapy on glandular cystitis. Methods The clinical data of glandular cystitis of 54 cases were retrospectively analyzed, all cases were treated individually and the applications of chemotherapeutic drugs bladder instillation were not adopted. Results Follow-up period ranged from 2.5 to 9.0 years, and the average was 4.6 years. Lower urinary tract symptoms score before treatment, 3 month , 6 month, 2 years , 4 years after treatment was (8.5 ± 3.7), (5.7 ± 2.3), (3.9 ± 1.3), (4.0 ± 1.9), (4.2 ± 1.9) scores, and the scores after treatment were improved significantly compared with that before treatment (P < 0.05). Recurrence rate 3 months , 6 months, 2 years and 4 years after treatment was 3.7％(2/54), 13.0％(7/54), 5.6％(3/54) and 1.9％(1/54). Conclusions Eliminating the inducements and improvement of symptoms provides a significantly curative effect in glandular cystitis. Postoperative bladder instillation chemotherapy is not recommended.

Objective To characterize contemporary electrographic neonatal seizures by video electroencephalogram (VEEG) and to assess the value and the limitations of two-channel (C3-C4/T3-T4) amplitude-integrated electroencephalogram (aEEG) plus original EEG signals used to diagnose neonatal seizure with video EEG as a golden standard.Methods Sixty-six neonates admitted to Children's Hospital of Fudan University from January 2011 to July 2011 with clinical or suspected clinical seizure were investigated and bedside VEEG were recorded for more than 3 hours.VEEG signals were transformed into three kinds of aEEG signals by Galileo NT PMS software:one-channel aEEG (C3-C4),one-channel aEEG (C3-C4) plus original EEG,two-channel aEEG (C3-C4/T3-T4) plus original EEG.Electrical seizure activity on VEEG was signed out with respect to its occurrence,duration and localization of seizure onset; while aEEG seizure was recorded only with its occurrence.The relationship between aEEG and VEEG was analyzed by Spearman analysis.The value and the limitations of aEEG to diagnose neonatal seizure were evaluated by sensitivity,specificity,positive predictive value and negative predictive value.Results A total of 62 traces were suitable for analysis.(1) VEEG showed 39 seizure activities,among which 8 status epilepticus; and the rest 31 neonates had 352 non-status epilepticus electrical seizures,79.3％ (279/352) of which occurred over the centrotemporal region.(2) Eight cases with status epilepticus on VEEG were all diagnosed as status epilepticus on aEEG.For non-status epilepticus electrical seizures,the sensitivity of aEEG for detection of electrical seizures was as followed:49.1％ (173/352) for one-channel aEEG,54.5 ％ (192/352) for one-channel aEEG plus original EEG,81.2％ (286/353) for two-channel aEEG plus original EEG.Results from one-channel aEEG,one-channel aEEG plus original EEG and two-channel aEEG plus original EEG were all related to VEEG (ρ =0.790,0.907 and 0.953,respectively,P＜ 0.01).(3) Sensitivity of seizure detection was 66.7％ (26/39,95％ CI:0.62-0.81) for one-channel aEEG,74.4％(29/39,95％ CI:0.78-0.96) for one-channel aEEG(C3-C4) plus original EEG and 89.7％ (35/39,95％ CI:0.89-1.00) for two-channel aEEG(C3-C4/T3-T4) plus original EEG.Conclusions VEEG might help aEEG in diagnosis of neonatal seizure.two-channel aEEG (C3-C4/T3-T4) plus original EEG could significantly increase the sensitivity of neonatal seizures indentification.

The study was aimed to investigate the effect of quercetin, flavonoid molecules on reversing leukemia multidrug resistance and its mechanism. K562/A cells were cultured in vitro with different concentrations of quercetin. Cell growth inhibition and adriamycin (ADR) sensitivity were detected by MTT method. Intracellular ADR concentration was determined by flow cytometry. Cell apoptosis was assayed by Annexin V/PI staining method. The expressions of drug transporter and apoptosis related genes were measured by real-time PCR array. The results indicated that quercetin inhibited the proliferation of K562 and K562/A in 5-160 µmol/L and with dose-dependent manner. Quercetin increased the sensitivity of K562/A cells to ADR in a low toxicity concentration. Flow cytometry showed that the quercetin increased the accumulation of ADR in K562/A cells when cells were co-cultured with 5 µmol/L ADR for 2 hours. Quercetin could induce the apoptosis of K562 and K562/A cells with dose dependent manner. Furthermore, some drug transport related genes such as ATP-binding cassette (ABC) and solute carrier (SLC) and some apoptosis-related genes such as BCL-2, tumor necrosis factor (TNF), tumor necrosis factor receptor (TNFR) families were down-regulated by quercetin. It is concluded that quercetin reverses MDR of leukemic cells by multiple mechanisms and the reversing effect is positively related to drug concentration.
Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans ; K562 Cells ; Quercetin ; pharmacology

Objective To discuss the long term clinical effect of ileal orthotropic neobladder.Methods From 1991 to 1998,79 patients,mean age 55(41～75)years,male 74,female 6,were followed up.The serum creatinine and urea,electrolytes,blood routine,B ultrasonic scan of the neobladder residual urine and IVU or MRU of the patients were followed up.The max transverse diameter of renal pelvis and the max verticaI/level diameter of neobladder were measured in 5,10 to 14,15 years of postoperative when IVU or MRU.All results of different time were compared by the multiple comparisons.The local or distant cancer recurrence and the complications of the operation Were evaluated. Results Sixty-four cases,58 male,6 famle,were long term followed up:mean time was 167 (range,121～216)months.Seven cases died of other diseases.Seven cases had pelvic recarrence.Two cases had urethral recurrence.Three cases died of tumor metastasis.One case had ureter recurrence.Forty-eight patients were alive more than 10 years.The value of the serum creatinine,urea,electrolytes and bloods routine of the patients were normal after 5,10 to 14 and 15 vears postoperative (P＞0.05).The max transverse diameter of the renal pelvis in 5,10 to 14 and 15 years Dostoperative were 14.0 mm,14.1 mm and 13.7 mm,respectively,P＞0.05.The max vertical/level diameter of the neobladder in 5,10 to 14,15 years of postoperative were 110.4 mm/90.4 mm,111.5 mm/95.3mm and 127.0 mm/97.0 mm,respectively,P＞0.05.The residual urine of 5 cases was more than 50 ml and had not increased during follow up.Eight cases with neobladder stone were cured by the intracavitary lithothrypsis.Two cases with uretheral stricture were cured by the intracavitary therapy.Twelve cases of 14 cases with inguinal hernia were cured by reoperation,2 cases accepted conservative treatment.Only 17 cases had no complication involve of the cancer and the operation. Conclusion The upper urinary tract and neobladder of the ileal orthotopic neobladder could be stable for long time,the cure rate of tumor is satisfactory and the lifetime follow up is necessary.

OBJECTIVETo study the correlation of homocysteine (Hcy) in plasma with nitric oxide synthetase (NOS) and endogenous carbon monoxide (CO) in the penile corpus cavernosum of type 2 diabetic rats.

METHODSThis study included 40 male Wistar rats, 10 as controls (Group A) and the other 30 as diabetes mellitus (DM) models. Four weeks after the model establishment, the model rats were divided into a DM group (Group B, n = 10), an insulin treated group (Group C, n = 10), and a folic acid and vitamin B12 treated group (Group D, n = 10). All the rats were injected with apomorphine and observed for penile erection at 8 and 12 weeks, and the levels of total plasma Hcy (tHcy), NOS and CO in the penile corpus cavernosum were measured at 12 weeks.

RESULTSCompared with Group A, the level of tHcy was significantly increased, while NOS and CO activities in the penile cavernous tis-sue and erectile function remarkably decreased in Group B (P < 0.01). The incidence rate of high Hcy was 55% in the DM rats. In comparison, the level of tHcy was obviously decreased, and the NOS activity and erectile function markedly increased in Groups C and D (P < 0.01). The Hcy level showed a significant negative correlation with NOS activity (rA = -0.89, rB = -0.76, rc = -0.91, rD = -0.91) and CO content (TA = -0.82, r, = -0.77, rc = -0.93, rD = -0.81).

CONCLUSIONHigh plasma Hcy can decrease NOS and CO activities in the penile corpus cavernosum, and consequently induce erectile dysfunction in DM rats, while insulin, folic acid and vitamin B12 can improve their penile erectile function by increasing NOS and CO activities.

Animals ; Carbon Monoxide ; metabolism ; Diabetes Mellitus, Experimental ; blood ; physiopathology ; Diabetes Mellitus, Type 2 ; blood ; physiopathology ; Folic Acid ; pharmacology ; Homocysteine ; blood ; Insulin ; pharmacology ; Male ; Nitric Oxide Synthase ; metabolism ; Penis ; drug effects ; metabolism ; Rats ; Rats, Wistar ; Vitamin B 12 ; pharmacology

This study was aimed to investigate the specific anti-leukemia cell effect of cytotoxic T lymphocytes (CTLs) induced by HL-60 or K562 cell-sensitized dendritic cells (DCs) from umbilical cord blood. 12 units of human umbilical cord blood (UCB) were collected and the mononuclear cells (MNCs) were isolated from UCB, then cultured with granulocyte monocyte colony- stimulating factor (GM-CSF), interleukin 3 (IL-3), recombinant human stem cell factor (SCF) and EPO for 3 - 4 weeks. Flow cytometry was used to determine the number of DCs and cell surface antigens before and after culture with monoclonal antibodies including CD83, CD1a, CD11c and CDw123. HL-60 and K562 were frozen-thawed, and released their tumor antigen peptides (TAP). The CTLs were produced by sensitizing T lymphocytes with DC-loaded HL-60 and K562 cell antigens. The test of (3)H-TdR incorporation was used to detect the immunostimulation activity of DCs. MTT assay was applied to evaluate specific cytotoxicity of CTL on leukaemia cells. The results indicated that the MNCs of UCBs cultured with GM-CSF, IL-3, EPO and SCF were shown to differentiate into CD1a(+) CD11c(+) CD83(+) CDw123(+) DCs. Numbers of DCs from UCBs remarkably increased in 2 - 4 weeks and then decreased. After culture with cytokines DCs increased (10.6 - 28.2) x 10(5)/ml in actual numbers. The CTL induced by DC pulsed with HL-60, K562 frozen-thawed lysates were effective to kill HL-60 and K562. Cytotoxicity of CTL to HL60 and K562 were (42.04 +/- 8.46)% and (31.25 +/- 11.07)% respectively. It is concluded that the MNCs of UCBs cultured with cytokines of GM-CSF, SCF, EPO and IL-3 can differentiate into CD1a(+), CD83(+), CD11c(+) and CDw123(+) DCs. The CTL induced by DCs pulsed with HL-60, K562 frozen-thawed lysates can effectively kill HL-60 and K562. These DCs as antigen presenting cells play an important role in cancer immunotherapy.
Dendritic Cells ; cytology ; immunology ; Fetal Blood ; cytology ; immunology ; HL-60 Cells ; Humans ; K562 Cells ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo construct a recombinant adenovirus vector for SH2-DED fusion gene and assess its inhibitory effect on the proliferation of K562 cells.

METHODSSH2-DED fusion gene and its mutant SH2mt-DED were amplified by splicing PCR and cloned into pAdTrack-CMV plasmid separately to construct the shuttle plasmids pAdT-SD-EGFP and pAdT-SmD-EGFP, respectively. After Pme I digestion, the shuttle plasmids were transformed into ultra-competent pAd5F35-BJ5183 cells to generate defective adenovirus vectors pAd5F35-SD-EGFP and pAd5F35- SmD-EGFP by homologous recombination. The vectors, linearized by Pac I digestion, were further transfected into AD293 cells for packaging and amplified by infecting AD293 cells repeatedly. K562 cells were then infected by the recombinant adenoviruses and the expression of SD was detected by Western blotting. MTT assay and flow cytometry were used to investigate the effect of Ad5F35-SD-EGFP and Ad5F35-SmD-EGFP on the proliferation of K562 cells.

RESULTSThe recombinant adenovirus vectors pAd5F35-SD-EGFP and pAd5F35-SmD-EGFP were constructed correctly, with a titer reaching 1.5×10(12) pfu/ml after amplification. Western blotting demonstrated that the target proteins were effectively expressed in transfected K562 cells. MTT assay and flow cytometry showed that transfection with pAd5F35-SD-EGFP resulted in growth inhibition rate of 55.21% in K562 cells, significantly higher than the inhibition rate of 17.95% following transfection with pAd5F35- SmD-EGFP and 7.33% following PBS treatment (P<0.05).

CONCLUSIONThe recombinant adenovirus vector Ad5F35-SD-EGFP we constructed can significantly inhibit the proliferation of K562 cells in vitro.

Adenoviridae ; genetics ; metabolism ; Apoptosis Regulatory Proteins ; biosynthesis ; genetics ; Cell Proliferation ; drug effects ; Cloning, Molecular ; Genetic Vectors ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; pathology ; Mutant Proteins ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Repressor Proteins ; biosynthesis ; genetics ; Shc Signaling Adaptor Proteins ; biosynthesis ; genetics ; Transfection

Objective:To investigate the epidemic characteristics of influenza in children and the features of severe influenza.Methods:From January 2016 to September 2022, 1 600 samples from hospitalized cases of severe acute respiratory tract infection and 7 660 samples from outpatients with influenza-like illness were collected. Influenza virus was detected by real-time RT-PCR. Other respiratory viruses in the samples of severe hospitalized cases and some samples of outpatients were detected. Clinical features of influenza virus infection and co-infection were analyzed.Results:The positive rate of influenza virus in the 1 600 hospitalized cases of severe acute respiratory infection was 6.63% (106 cases). H1N1, H3N2, BV and BY were deteted in 49.06% (52 cases), 17.92% (19 cases), 29.25% (31 cases) and 3.77% (4 cases) of the 106 cases, respectively. The positive rate of influenza virus in the 7 660 out-patient cases was 15.01% (1 150 cases), and H1N1, H3N2, BV and BY were detected in 22.17% (255 cases), 30.96% (356 cases), 41.39% (476 cases) and 5.48% (63 cases) of the infected cases, respectively. Influenza A (H1N1) virus was more likely to cause severe influenza in children (χ 2=37.978, P<0.001), while seasonal H3N2 and BV strains were less likely to cause severe influenza in children (χ 2=7.871, P=0.005; χ 2=5.948, P=0.015). There was no statistically significant difference in the positive rates of BY lineage in the two groups. Severe influenza mainly occured in the peak season of influenza epidemic. There was no significant difference in the clinical manifestations between the children infected with the four different influenza viruses. In the 106 severe cases of influenza, the co-infection rate of influenza virus with other respiratory viruses was 17.92% (19 cases), while the co-infection rate reached 34.81% (47 cases) in 135 outpatient cases of influenza. The difference in the co-infection rates was statistically significant between outpatient and hospitalized cases (χ 2=10.734, P=0.001). Conclusions:Influenza A (H1N1) virus was more likely to cause severe influenza in infants and young children in comparison with seasonal H3N2 and BV. There was no significant difference in the clinical features of influenza caused by H1N1, H3N2, BV and BY. Co-infection of influenza virus with other respiratory viruses is not a major risk factor for severe influenza in infants.

OBJECTIVETo investigate the effect of gene Af116609 on gastric cancer multi-drug resistance (MDR) by introducing it into gastric cancer multi-drug resistant (MDR) cell line SGC7901/VCR.

METHODSGene Af116609 was cloned from SGC7901/VCR by RT-PCR and its differential expression between gastric cancer MDR cells and its parental cells was displayed by Northern blot. The gene was introduced to gastric cancer cells by transfection of recombinant eukaryotic expression vector by electroporation. MTT assay in vitro was applied to investigate its effect on multi-drug resistance phenotype of gastric cancer cells.

RESULTSThe full length CDS of gene Af116609, as long as 327 bp, was cloned from gastric cancer MDR cell line SGC7901/VCR and its sequence was coincident with the hypothetical gene Af116609 in GenBank. It was overexpressed in MDR cells than its parental cells at mRNA level. In the MTT assay in vitro, the drug sensitive cells transfected with sense eukaryotic expression vector showed upregulated targeted gene, with increased resistance to vincristine, 5-fliorouracil and arabinoside, and decreased resistance to adriamycin, but no influence on resistance to methotrexate. However, the drug resistant cells transfected with anti-sense eukaryotic expression vector, showed down regulated targeted gene, with less resistance to all the five anticancer drugs to different degrees.

CONCLUSIONGene Af116609 is related to MDR phenotype of gastric cancer cells and may become a candidate molecular target to reverse the MDR of gastric cancer.

Antineoplastic Agents, Phytogenic ; pharmacology ; Autoantigens ; genetics ; Cell Line, Tumor ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Humans ; RNA, Small Cytoplasmic ; genetics ; Ribonucleoproteins ; genetics ; Stomach Neoplasms ; genetics ; pathology ; Vascular Endothelial Growth Factor A ; biosynthesis ; Vincristine ; pharmacology