Dendritic cells (DC) are important antigen-presenting cells in the development of anti-leukemic T-cells responses and it' s reported that DC can differentiate from monocytes or granulocytes in vitro. In the present study, the human leukemic DC were generated from acute promyelocytic leukemic( APL)cells after treatment with cytokines. M3 APL cells were isolated from peripheral blood of patients and incubated with rhGM-CSF( 100ng/ml)or rhGM-CSF( lOOng/ml) plus rhIL-4(500U/ml)for 14 days and the cells were co-cultured wirh TNF-?(100ng/ml)during last 3 days. The results demonstrated that both proliferation and differentiation of APL cells were induced by GM-CSF as the number of cultured cells increased and the cells expressed high level of CD45. Some cells displayed the characteristics of monocytes which expressed CD14 and some were leukemic DC as they expressed GDI a. The co-culturation of APL cells with GM-CSF and TNF-? augmented the differentiation of cells and about 35 percent of leukemic DC generated (was obtained). The maturation of APL cells could also be induced by GM-CSF plus IL-4 but there were few monocytes generated and about 10 percent of the cells were leukemic DC. If cultured for 3 weeks, the number of leukemic DC increased to 60 percent. The addition of TNF-? could augmented GM-CSF and IL-4 induced maturation of cells significantly and 90 percent of leukemic DC generated. Analysis of the cells with electric microscopy indicated that these leukemic DC displayed the similarly morphologic characteristics as monocyte-derived DC and there were still a few granules in their cytoplasm. The leukemic DC expressed high levels of HLA-DR, B7-1, B7-2 and CD54 molecules and could stimulate allo-T cells to proliferate in vitro. Such kind of leukemic DC might be useful tools for the generation of specific anti-tumor CTL and play important roles in immunotherapy of APL.

TNF gene was transfected into murine LAK cells by retrovirus. Our results showed that TNF gene-transfected LAK cells secreted TNF more than normal LAK cells and control gehe-transfected LAK cells. The in vitro growth ability and cytotoxicity of TNF gene-transfected LAK cells were augmented significantly.The cytotoxicity of ,TNF gene-transfected LAK cells was markedly inhibited by anti - TNF monoclonal antibody, indicating that the, above augmentation was mediated by TNF secreted by transfected LAK cells. Significant therapeutic effect on the ascitic liver carcinoma.-bearing mice was achieved by i.p. injection of low dosage TNF gene transfected LAK cells and IL - 2.

Peripheral blood mononuclear cells (PBMNC) isolated from patients with acute leukemia (AL) and from normal controls were cultured in medium containing 1000 units/ml of recombinant interleukin 2 (IL-2). Marked LAK activity was induced on the third culture day in the normal controls,, with the highest cytotoxicity appearing between day 3 and 5 whereas induction of LAK activity in the AL patients began on the 5th day of culture, with the peak level appearing at day 15, showing that the peak of LAK activity was significantly delayed in AL. LAK cells surface phenotyping tests showed that CD8+ and CD16+ positive cells began to increase significantly from day 5 and reached the highest level at week 3, whereas CD4+ subclass began to decrease on day 5 and dropped to the nadir at week 3. The proportion of CD8+ and CD16+ cells were positively cor related with LAK activity, but, that of the CD4+ cell was inversely related with the LAK activity.

Objective:To observe the expression of ATPase F1? in colorectal cancer(CRC) tissues and in LoVo cells,and to discuss its clinical significance.Methods:Expression of ATPase F1? protein in 44 CRC specimens and their adjacent normal tissues(August 2007 to December 2007,Changhai Hospital) and ATPase F1? mRNA in 8 colorectal cancer tissues and their adjacent normal tissues were examined by immunohistochemistry EnVision assay and RT-PCR,respectively.Expression of ATPase F1? on the cell surface of LoVo cells was observed by immunofluorescence.The inhibitory effect of anti-ATPase F1? antibody on the proliferation of LoVo cells was evaluated by CCK-8 assay.Results:Expression of ATPase F1? in the 44 CRCs were significantly higher than those in the adjacent normal tissues as detected by immunohistochemistry(P0.05).Expression of ATPase F1? was observed on the cell surface of LoVo cells,and anti-ATPase F1? antibody significantly inhibited the proliferation of LoVo cells(P

Abstract: This study is to elucidate the metabolic pathway of 1,2-[bis (1,2-benzisoselenazolone-3 (2H)-ketone)]-ethane (BBSKE) in rats. Rats were administrated with a single dose of BBSKE 200 mg x kg(-1). The metabolites in rat urine, feces, bile and plasma were identified by LC-MSn analysis. The characterization of fragment ions from LC-MSn chromatography and mass spectrometry was applied to the investigation of structures of metabolites. Three phase I metabolites were detected in rat urine and feces. Two of them were also found in plasma and one existed in bile. These products were derived from oxidized, methylated and S-methylated BBSKE, separately. One phase II glucuronide of BBSKE was also found in bile. Therefore, it is possible that BBSKE was metabolized by oxidization, methylation and glucuronidation.

Objective To observe the clinical therapeutic effect of Xuebijing injection on vascular endothelial cell ( VEC) injury,microcirculation disorder and organ dysfunction in patients with sepsis. Methods Seventy?three patients with sepsis were randomly divided into two groups:Xuebijing injection?treated group (40 cases) and control group (33 cases). Routine medicine therapy was applied in both groups. Additionally, the Xuebijing injection?treated group was treated with Xuebijing injection 100 ml and saline 100 ml by intravenous drip every 12 hours for consecutive 5 days. Vascular endothelial injury index, including soluble thrombomodulin( sTM) ,vascular endothelial growth factor 2 ( VEGF?2) ,endothelial specific molecule 1 ( ESM?1),microcirculation index of arterial blood lactic acid (Lac),central venous oxygen saturation (ScvO2),total microvessel density (TVD),the perfusion vascular density (PVD),proportion of perfused vessels (PPV) and microvessel flow index ( MFI) of the two groups before and after therapy were observed and the sequential organ failure score ( SOFA) was recorded before and after treatment. Results Senventy?three patients with sepsis had different degrees of increase in vascular endothelial damage markers,lactate and sequential organ failure scores in arterial blood, while the central venous blood oxygen saturation ( ScvO2 ) , the total vascular density of the sublingual microvasculature ( TVD) ,perfused vessel density ( PVD) ,proportion of perfused vessels ( PPV) and microcirculatory flow index ( MFI) decreased before treatment. After 5?day treatment,the above indicators of all patients were improved,the indexes in the Xuebijing injection group decreased significantly,compared with the control group ,sTM ( (16. 91±4. 55) μg/L,(19. 51±4. 09) μg/L,t=-6. 021,P<0. 05),VEGF?2 (50. 8 (17. 8,127. 7) ng/L vs. 74. 9(22. 7,155. 1) ng/L,t=4. 227,P<0. 05),ESM?1 ( (10. 20 ±2. 43) μg/L vs. (14. 80±3. 52) μg/L,t=-4. 113,P<0. 05),Lac( (2. 1±0. 7) mmol/L vs. (3. 7±1. 1) mmol/L,t=2. 366,P<0. 05) and SOFA ( (5. 9±2. 1) vs. (8. 7±2. 6),t=-7. 990,P<0. 05). ScvO2( (0. 771±0. 153) % vs. (0. 641±0. 113) %,t=5. 061,P<0. 05),PVD ( (16. 8±6. 1) mm/mm2 vs. (12. 1±5. 1) mm/mm2,t=4. 002, P<0. 05),PPV ( (66. 2±21. 3) % vs. (50. 4±19. 3) %,t=-2. 550,P<0. 05) and MFI (6. 2 ±2. 4) vs. (3. 8 ±2. 2),t=-5. 001,P<0. 05) were significantly higher than those in the control group in the same period. sTM and PPV had a significant negative correlation (r=-0. 755,P=0. 000),PVD,PPV,ESM?1 and MFI were negatively correlated (r=-0. 665,P=0. 000; r=-0. 600,P=0. 000; r=-0. 469,P=0. 000),PPV,MFI and SOFA were negatively correlated ( r=-0. 798,P=0. 000;r=-0. 995,P=0. 000);sTM,ESM?1 and SOFA were significantly positively correlated ( r = 0. 883, P = 0. 000;r = 0. 881, P = 0. 000 ) . Conclusion Vascular endothelial cell dysfunction probably plays an important role in the pathophysiology of sepsis and Xuebijing injection has therapeutic effect on sepsis by protecting vascular endothelial cell function.

Objective To explore the effect of different transcranial pulse current stimulation(tPCS)program on the elimination of fatigue in physical cognitive mixed tasks.Methods Thirty healthy college athletes were randomly divided into group Ⅰ'and groupⅡ',each of 15.Then,both groups of sub-jects exercised on power bicycles.Ten subjects were screened from Group Ⅰ'and Ⅱ'meeting the crite-ria of moderate and severe physical cognitive mixed task fatigue using the Rating of Perceived Exer-tion respectively.Both groups received five tPCS interventions.Before and after each intervention,the subjects were tested for heart rate variability(HRV)and oxygenated hemoglobin(HbO2)concentration,and the effect of different stimulations on the fatigue elimination in physical cognitive mixed tasks of different severities were measured.Meanwhile,the HRV measurements included root mean square of difference between adjacent R-R intervals(RMSSD),standard deviation of all normal-to-normal inter-vals(SDNN),high frequency(HF)and low frequency(LF).Results①After tPCS intervention,the aver-age SDNN,HF and HbO2 increased significantly(P<0.05),while the average IF decreased significantly(P<0.05).②In physical cognitive mixed task of moderate fatigue,the biggest change of each index ap-peared after the tPCS program D(20 min,sensory intensity).③In the physical cognitive mixed task of severe fatigue,the change range of each index was the largest after the tPCS program C(20 min,sensory intensity + 0.2 mA).Conclusion ①After physical cognitive mixed tasks,different tPCS stimu-lation programs have different effects on the elimination of fatigue with an optimal"stimulant dose".②The effects of five intervention programs on the elimination of physical fatigue of athletes are as fol-lows:For the elimination of moderate fatigue of physical cognitive mixed task,program D of tPCS(20 min,sensory intensity)has the greatest effect,while for the elimination of severe fatigue of physical cognitive mixed task,program C of tPCS(20 min,sensory intensity + 0.2 mA)has the greatest effect.

Objective:To evaluate the efficacy of ultrasound-guided superior laryngeal nerve block(SLNB) combined with intravenous anesthesia for improving pediatric fiberoptic bronchoscopy.Methods:Forty pediatric patients of either sex, aged 3-6 yr, of American Society of Anesthesiologists Physical Status classificationⅠor Ⅱ, with body mass index of 18-24 kg/m 2, undergoing fiberoptic bronchoscopy in Cangzhou Central Hospital in 2022, were divided into 2 groups ( n=20 each) by a random number table method: ultrasound-guided SLNB plus intravenous anesthesia group (group A) and topical anesthesia plus intravenous anesthesia group (group B). After sedation with dexmedetomidine and esketamine, ultrasound-guided bilateral SLNB was performed with 1% lidocaine 0.5 ml (for each side)in group A, and topical anesthesia was performed with 1% lidocaine in nasal and pharyngeal cavities in group B. After completion of the surgery procedure, propofol was continuously infused at 5 mg·kg -1·h -1 until completion of diagnosis and treatment. An increment of propofol 1 mg/kg was intravenously given if severe bucking or body movement occurred during operation. Mean arterial pressure (MAP), heart rate (HR) and SpO 2 were recorded on admission to the operating room (T 0), immediately after sedation (T 1), immediately after bronchoscopy entering the glottis (T 2), 5 min after start of treatment (T 3) and at the end of examination (T 4). The occurrence of intraoperative hypoxemia, HR <60 bpm, and MAP <50 mmHg were recorded, and the additional dose of propofol was recorded. The venous blood samples were collected at T 0 and T 4 to determine plasma cortisol concentrations by chemiluminescence.The surgeon′s satisfaction score was recorded. The complications of SLNB were also recorded within 2 h after operation in group A. Results:Compared with group B, HR was significantly decreased at T 2 and T 3, SpO 2 was increased, the intraoperative additional dosage of propofol and incidence of hypoxemia were decreased, and the surgeon′s satisfaction score was increased, and the concentrations of cortisol were decreased at T 4 in group A ( P<0.05). No HR<60 bpm and MAP<50 mmHg were found in two groups. No SLNB-related complications were observed after operation in group A. Conclusions:Ultrasound-guided SLNB combined with intravenous anesthesia is safer for pediatric fiberoptic bronchoscopy and can improve the analgesic effect and is more helpful in inhibiting intraoperative stress responses when compared with conventional anesthesia.

Objective: To investigate the etiology composition of enterovirus (EV) in patients with severe hand, foot, and mouth disease (HFMD) in children. To assess the diagnostic value of cerebrospinal fluid (CSF) tests in severe HFMD, and to find the key laboratory tests for severe HFMD. Methods: A total of 288 hospitalized cases of children clinically diagnosed with severe HFMD in Hangzhou Children′s Hospital were included from March to July 2016. Throat swabs were collected and enterovirus nucleic acids were detected by fluorescence quantitative reverse transcription (RT)-PCR. Synchronous CSF and serum samples were collected for EV-A71 and CV-sackievirus A16 (CV-A16)-IgM antibody detection. CSF samples underwent routine and biochemical tests. Normally distributed continuous variables were compared using t test. Non-normal distribution continuous variables were compared using Mann-Whitney U test. Differences between categorical variables were compared with χ² test. Results: The total positive rate of enterovirus nucleic acid EV-A71/CV-A16/EV by fluorescence quantitative RT-PCR in the 288 cases of children clinically diagnosed with HFMD was 83.7% (241/288), including EV-A71 55.2% (159/288), CV-A16 4.9% (14/288) and the other enterovirus 23.6% (68/288). Among the other enterovirus group, there were 29.4% CV-A6 (20/68) , 16.2% CV-A4 (11/68) and CV-A10 10.3% (7/68). The total positive rate of combined serum and CSF detection of EV nucleic acid and EV-A71 and CV-A16 IgM antibody was 98.3% (283/288). EV nucleic acid positive rate was 83.7% (24/288). The positive rates were statistically different (χ² =37.289, P=0.000). The CSF nucleated cells count in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (Z=-4.472 and -9.991, respectively, both P<0.05). The CSF nucleated cells positive rate in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (χ²=43.857 and 133.078, respectively, both P<0.05). The CSF protein level in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (Z=-3.151 and -5.255, respectively, both P<0.05). The CSF protein positive (>400 mg/L) rate in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (χ²=4.956 and 11.795, respectively, both P<0.05). The CSF nucleated cell counts and positive rates in EV-A71 IgM antibody-positive subgroup were both higher than those in antibody-negative subgroup (both P<0.05). The CSF protein level and elevated proportion in antibody-positive subgroup were both higher than those in antibody-negative subgroup (both P<0.05). The lactate dehydrogenase concentration in antibody-positive subgroup was significantly higher than those in antibody-negative subgroup (P<0.05). The EV-A71 IgM antibody in serum was significantly correlated with the antibody in CSF (r=0.600, P<0.05). Conclusions EV-A71 is still the most important pathogen of severe HFMD in Hangzhou in 2016. Other enterovirus such as CV-A6, CV-A4 increases compared to those in 2014 and 2015. The CSF routine and biochemical tests and the IgM antibody levels can serve as an important indicator for the diagnosis of children with severe HFMD.