Objective To investigate the expression of Toll-like receptor 4 (TLR4) and myeloid derived suppressor cells(MDSC) in bone marrow cells in children with acute myeloid leukemia (AML),and to detect its relationship with the clinical features,the effect of chemotherapy and prognosis.Methods Twenty-nine cases of children with AML were collected from June 2013 to March 2014 in People's Hospital of Wuhan University,in which 11 cases of low-risk group,10 cases of middle-risk group,8 cases of high-risk group;and 17 cases of non blood disease was as the control group.The expressions of TLR4 and MDSC were detected by using reverse transcription-polymerase chain reaction (RT-PCR),Western blot methods,immunohistochemical staining,and flow cytometry,respectively,in the bone marrow cells of 29 children with AML.Results The mRNA and protein expression of TLR4 in the initial treatment group was higher than those in the complete remission group(t =3.092,3.393,all P ＜ 0.05).The mRNA and protein expression of TLR4 in the relapse group was higher than those in the complete remission group(t =4.013,4.279,all P ＜ 0.05).The positive expression rates of MDSC in the above 3 groups were (29.77 ± 1.39) ％,(5.19 ± 0.65) ％,(38.62 ± 3.54) ％,respectively,compared with the control group [(1.32 ± 0.27) ％] and there was significant difference(all P ＜0.05).The positive expression rates of TLR4 and MDSC in the initial treatment group,relapse group and complete remission group were significantly higher than those in the control group,with significant differences (initial treatment group TLR4:t =3.559,P ＜ 0.05;MDSC:t =3.727,P ＜ 0.05;relapse group TLR4:t =4.043,P ＜ 0.05;MDSC:t =4.125,P ＜ 0.05;complete remission group TLR4:t =2.798,P ＜ 0.05;MDSC:t =3.469,P ＜ 0.05).Pearson rank correlation analysis showed that there was a positive correlation between the expression of TLR4 and MDSC (r =0.673,P ＜0.01).Conclusions The expressions of both TLR4 and MDSC play an important role in onset,progression,curative effect and prognosis in children with AML,and the two may play an importment role in synergistic effect.

Objective To investigate the molecular mechanism underlying lymphocyte functionassociated antigen-1 (LFA-1) / intercellular adhesion molecule-1 (ICAM-1) mediated anti-neoplastic effects of cytokine induced killer (CIK) cells. Methods Lymphocytes isolated from peripheral blood of children leukemia were induced with interferon-gamma (IFN-y), anti-CD3 monoclonal antibody (CD3McAb) and interleukin-2 (IL-2) and co-cultured with dendrite cells (DC) to generate DC-CIK cells. When treated with LFA-1 monoclonal antibody, cytotoxicity of DC-CIK cells against leukemia cell lines was measured by the MTT assay, while RT-PCR and Western blotting were used to determine mRNA and protein expressions of GATA-3 and T-bet in DC-CIK cells, respectively. IL-12, IFN-γ and tumor necrosis factor-α (TNF-α) levels released by DC-CIK cells were quantified by ELISA. Results Induced DC-CIK cells were regular, round and transparent with variable cell volume and cellular aggregation. When treated with mouse anti-human LFA-1 monoclonal antibody, the cytotoxicity decreased mostly towards B95 cells under administration of 20 μg/ml LFA-1 monoclonal antibody in comparison with the control group(t =10.138, P ＜0.05). It led to a highest elevation of GATA-3 mRNA and protein levels (t =16.386, P ＜ 0.05; t =22.652, P ＜ 0.05) and a most decrease of T-bet mRNA and protein levels (t =17.728, P ＜0.05; t =17.452, P ＜0.05) under 20 μg/ml LFA-1 monoclonal antibody in B95 cells group in comparison with the control group. The expression levels of IL-12,IFN-γ, and TNF-o in supernatant were the lowest under 20 μg/ml LFA-1 monoclonal antibody in B95 cells group in comparison with the control group (t =21.621, P ＜0.05; t =13.739, P ＜0.05; t =15.278, P ＜0.05).Conclusion GATA-3 and T-bet were implicated in the LFA-1/ICAM-1 mediated anti-neoplastic effects of DC-CIK cells via activation of the Th1 pathway, with high secretion of Th1 cytokines, such as IL-12, IFN-γ and TNF-α.

Objective To explore the feasibility and advantages of transvaginal myomectomy(TVM).Methods A total of 65 patients with uterine myoma were treated with TVM from August 2002 to August 2004.The myomectomy was performed through a vaginal incision,which was transversely made through anterior or posterior fornix of the vagina.The uterus was exposed outside the incision with a pulling suture.The muscular layer covering the myoma was incised and the myoma was removed.Then the incisions of the uterus and vagina were closed respectively by using absorbable sutures.Results The TVM was successfully completed in all the 65 patients.The surgical time was 25~140 min(mean,56 ?19 min),the postoperative bleeding amount was 60~650 ml(mean,170?45 ml),and the length of hospital stay after operation,2~5 d.A follow-up was carried out for 2~12 months(mean,3.6?2 months) in 58 patients.The flow of menstrual cycle recovered to normal levels in 40 patients and was less than normal levels in 2 patients.Pressure symptoms of adjacent organs disappeared,and no residual tumors were detected on B-ultrasonography.Conclusions Transvaginal myomectomy is a safe and reliable procedure with little invasion and quick recovery.

ObjectiveTo discuss the value of interrupted circular suture in hemostasis of placenta previa during cesarean section. Methods We summarized 54 caesarean section patients with placenta previa. Results The hemostasis was succeeded in all of the 9 patients and uterus was retained without postpartum complications. The duration of operation was obviously shorter than that of hysterectomy( P＜0.05). Bleeding and blood transfusion were less than that of hysterectomy, but without statistical difference (P＞0.05). ConclusionInterrupted circular suture is one of the efficient methods in controlling postpartum bleeding during caesarean section with placenta previa.

Apnea in premature infants is a common preterm neonatal serious disease, persistent apnea will cause harm to brain and organ development. Methylxanthine compounds such as theophylline and caffeine are used to treat apnea in premature infants. Among them, the caffeine treatment of greater safety and ease of administration once a day, and more attention due to caffeine safe, effective, cost-effective the best, Aranda described it as "silver bullet in neonatal medicine". Caffeine reduced the number of apneic episodes during the first 2 to 7 d and the number of intermittent positive pressure ventilation use and showed a dependency on other beneficial effects such as can reduced time of tracheal intubation and nasal catheter continuous positive airway pressure, reduce the duration of continuous oxygen supply, improve the success rate of weaning, reduce the incidence of bronchopulmonary dysplasia, reduce patent ductus arteriosus, reduce the use of hormones, retinopathy of premature children have a positive therapeutic effect. Caffeine treatment reduces positive airway pressure, thereby reducing overall treatment costs. In 2013, China approved caffeine citrate for the treatment of apnea in premature infants. In this paper, the pharmacokinetics of caffeine in premature infants and its application in apnea and related diseases of premature infants were reviewed.

Objective To study the effects of peripheral nervous system on early and middle periods of fracture healing. Methods Denervated tibia fracture model and innervated tibia fracture model were made at the same rat. The animals were executed at day 15 and day 30 after operation respectively. Callus sizes were evaluated by radiograph. The mechanical properties of calluses were recorded in a three point bending test. The callus microstructures were measured by bone histomorphometry.Results Radiograph showed an increased callus formation in the denervated group both on day 15 and day 30 after operation (P0.05). Compared with the innervated groups, 30 days dynamic parameters indicated that mineral appositional rates of the denervated sides significantly decreased (P

Objective To investigate the cytotoxicity and mechanism of killing tumor of cytokine-induced killer (CIK) cells in vitro.Methods Mononuclear cells were acquired freshly from bone marrow of children with leukemia,and the cells obstained were induced into dendritic cells by adding granulocyte-macrophage colony-stimulating,IL-4,TNF-? and other cytokines.Lymphocytes cells were isolated freshly from peripheral blood of children with leukemia by Ficoll-Hypaque density centrifugation,and the cells obstained were induced by IFN-?,IL-2 and CD3McAb.The DC cells and CIK cells were co-cultured for 10-25 days,then DC-CIK cells were obtained.Phenotypes of DC-CIK were analyzed by flow cytomery.The cytotoxicity of DC-CIK against a variety of leukemic cell lines was investigated by MTT technique.When treated with mouse-anti-human LFA-1 monoclonal antibody,the expression of GATA-3 and T -bet in the levels of mRNA and protein were mea-sured by using RT-PCR and Western Blot technique,respectively.Results In the first 0-6 days,DC-CIK induced slowly,the proliferation of DC-CIK got 100-fold at the 13th day,cells were rapidly proliferating in the first 13-21 days.The maximum proliferation of DC-CIK reached at the 22nd day.The phenotypes of CD3,CD11a,CD54,HLA-DR were expressed highly; CD3/CD56,CD25,CD28,CD69,FasL were expressed moderately on DC-CIK.The expression of CD16 was not increased.DC-CIK possessed the cytotoxicity against tumor cells of B95,Jhhan and M07e.The effect was stronger to B95,there was no significant difference when the efficiency target ratio was 12.5:1.0 or 25:1,the cytotoxicity reached about 50% and 60%,respectively,against tumor cells of B95.However,it was not obvious to Jhhan and M07e.When the efficiency target ratio was 12.5:1.0 or 25:1,the cytotoxicity reached to 27.21%,25.13%,33.05%,29.72%,respectively,against tumor cells of Jhhan and M07e.When treated with mouse-anti-human LFA-1 monoclonal antibody,the expression of GATA-3 in the level of mRNA was up-regulated(t=3.425,4.523 Pa

Objective: To establish the method of determination of the content of Berberine hydrochloride and Palmatine hydrochloride in Wuji Pills. Methods: Ultrasonic extraction, HPLC was used to determine Berberine hydrochloride and Palmatine hydrochloride in Wuji Pills on C 18 Column, using 0.033mol/L KH 2PO 4 CH 3CN(90∶40) as mobile phase, with detection at 265nm. Results: The linearity of this method was well. The average recoveries were 99.07% for Berberine hydrochloride and 98.59% for Palmatine hydrochloride. RSD were 1.3% and 1.7%, respectively. Conclusion: This method shows good separating degree and can be used for quality control of Wuji Pills.

Objective To investigate whether sex,age-of-onset,education and asymmetry affect non-motor symptoms (NMS)in Parkinson disease,and to analyze the relationship between motor symptoms and NMS in Parkinson's disease.Methods The detailed clinical information of 157 patients with idiopathic Parkinson's disease(PD)was recorded and followed up.The data was calculated with SPSS statistic software.Sex had an impact on sleeping disorder (P＜ 0.05).Sleep disorder in female was more prominent.Cognitive disorder was affected by age-of-onset and education duration.The pain and sensory disorder were affected by age-of-onset (P ＜ 0.05).Differences in the effects of asymmetry on NMS were not statistically significant (P＞ 0.05).The higher the scores of postural instability and gait difficulty (PIGD)were,the lower the Mini-Mental State Examination (MMSE)score was(β=-1.837,P =0.004).The higher the tremor score was,the higher the Hamilton Depression Scale(HAMD) score was(β=3.460,P =0.001).The higher the rigidity akinesia score was,the higher the autonomic dysfunction score was(β=0.104,P=0.006).Conclusions The non-motor symptoms of Parkinson's disease are affected by the age,sex and cultural level.Symmetryor-not does not affect Parkinson's disease.Non-motor symptoms and motor symptoms are closely linked.The encountered posture and gait abnormalities of patients should trigger physician alerts to focus on patients' cognitive function.The encountered tremor of patients should trigger physician alerts to focus on their emotional changes.Facing to the patient with rigidity and less moving,physician should alert to the occurrence of autonomic symptoms.

Objective To investigate the effect of three blood purification techniques on minerals disorder in maintenance hemodialysis patients,and provide clinical guidance for patients to choice the blood purification techniques.Methods Eighty-eight maintenance hemodialysis patients were divided into three groups according to the blood purification techniques:hemodialysis (HD) group (30 cases),hemodiafiltration (HDF) group (30 cases),and hemoperfusion (HP) group (28 cases).Serum urea nitrogen,creatinine,calcium,phosphorus,intact parathyroid hormone (iPTH),ftbroblast growth factor (FGF)-23 and so on before and after treatment were measured and compared among the groups.Results The serum phosphorus in HD group,HDF group and HP group before treatment were (1.93 ±0.44),(2.11 ±0.54) and (2.17 ±0.59)mmol/L,and after treatment were (1.01 ±0.23),(0.84 ±0.19),(0.99 ±0.27) mmol/L.There were significant differences in serum phosphorus level after treatment compared with that before treatment in the three groups (P ＜0.05).There were no significant differences in the descend rate of serum phosphorus among the three groups (P ＞0.05).There were no significant differences in clearance index of serum phosphorus among the three groups (P ＞ 0.05).The serum iPTH in HD group before treatment was (48.8 ± 42.9) pmol/L,and after treatment was (49.9 ± 42.9) pmol/L.The serum iPTH in HDF group and HP group before treatment were (64.7 ± 45.4) and (50.4 ± 45.9) pmol/L,after treatment was (46.2 ± 37.8) and (35.8 ± 36.5) pmol/L.There were significant differences in serum iPTH level after treatment compared with that before treatment (P ＜ 0.05).There were no significant differences in the descend rate and clearance index of serum iPTH in HDF group and HP group (P ＞ 0.05).There was no significant difference in serum FGF-23 in HD group before and after treatment (P＞ 0.05).The serum FGF-23 in HDF group and HP group before treatment were (782.5 ± 105.8) and (879.5 ±97.2) ng/L,after treatment were (712.0 ±98.1),(823.5 ± 89.1) ng/L.There were significant differences in serum iPTH level after treatment compared with that before treatment in HDF group and HP group (P ＜ 0.05).The descend rate of serum FGF-23 in HD group,HDF group and HP group were (5.7 ±2.8)％,(12.3 ±6.2)％ and (9.1 ±4.6)％,and there was significant difference among the three groups (P ＜0.05).The clearance index of serum FGF-23 in HD group,HD F group and HP group were 0.06 ± 0.05,0.19 ± 0.11 and 0.12 ± 0.08,and there were significant differences among the three groups (P ＜ 0.05).There were no significant differences in the descend rate and clearance index of serum urea among the three groups (P ＞ 0.05).Conclusions HD can only clear serum phosphorus.HDF and HP can clear serum phosphorus,iPTH and FGF-23 effectively,while HDF has better clearance effect on FGF-23.The HDF and HP blood purification can reduce minerals disorder in maintenance hemodialysis patients and has important clinical significance in improving the long-term prognosis of the patients.