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.

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.

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.

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 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 the protective effect of the extract of ginkgo biloba on myocardial tissue in aged rat.Methods The rats aged 20 months were given the extract of ginkgo biloba (EGB) and ALT-711 respectively by garage for 16 weeks.The aged controls and adult rats were infused with the same volume saline.The superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), malonyt diadehyde (MDA) in blood samples and advanced glycation end products (AGEs) in the myocardial tissue were measured .The myocardial histopathological changes under electron microscope and the mitochondrial DNA (mtDNA) deletion rate in myocardial cells were observed.Results Compared with the adult rats, the content of AGEs in myocardial cells in aged rats was significantly increased [(33.5±1.3)AU/mgHYP us.(18.1±1.2)AU/mgHYP, t= 7.18,P<0.05] and the levels of SOD and GSH-PX in blood samples were decreased [(138.4±3.8) U/mlvs.(227.7±13.8)U/ml, (1283.8±28.8) U/ml vs.(2114.1 ±135.9)U/ml, t=-19.59, -18.79;both P<0.01].The MDA level in the serum and mtDNA deletion rate in aged rats were higher than in adult rats[(6.7±0.6) mmol/ml vs.(4.1±1.0) mmol/ml, (0.18054±0.0718) % vs.( 0.0060±0.0001)%, t=7.18,6.98;both P<0.05].Compared with the aged controls, the content of AGEs in myocardial cells, the level of MDA and mtDNA deletion rate were significantly decreased in EGB and ALT-711 treatment adults (all P<0.05).The SOD and GSH-PX in blood samples were increased in EGB and ALT-711 treatment adults (all P<0.05).Conclusions Nonenzymatic glycation may play an important role in myocardial aging, which may be amplified by oxidative stress.EGB and ALT-711 may have the same anti-aging effects by inhibiting nonenzymatic glycation and oxidative stress.

Objective To explore the role of pregabalin on the expression of c-fos in spinal dorsal horn of the rat with neuropathic pain.Methods Thirty Wister rats (male) were divided into shamoperated group,model group and pregabalin group,with 10 cases in each group.The effect of the pregabalin on the heat pain threshold and expression of c-fos in spinal dorsal horn of the rat with neuropathic pain were observed.Results The heat pain threshold in model group at 3rd,4th,5th,6th,7th,10th and 14th day after operation was significantly lower than that in sham-operated group and pregabalin group at same time (P ＜ 0.05).The heat pain threshold in pregabalin group was lower than that in sham-operated group from 2nd day after operation,but there was no significant difference (P ＞0.05).At the 14th day after operation,the number of Fos-like-immunoreactivity (FLI) positive cells in sham-operated group,model group and pregabalin group was (16.4 ± 0.6),(66.7 ± 3.3) and (22.8 ± 1.5)cases,and the number of FLI positive cells in model group was significantly higher than that in shamoperated group and pregabalin group (P ＜ 0.05).Conclusions Pregabalin has analgesic activity on rat with neuropathic pain.Pregabalin activates spinal cord glial cells raisedby the injured peripheral nerve and infection,and also reduces the noxious stimulation of afferent pain to spinal dorsal horn neurons.

Objective To explore the expression of antiapoptotic gene Aven in childhood acute lymphoblastic leukemia (ALL) and its relationship with clinical features of ALL. Methods Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the expression of Aven mRNA in 55 cases of childhood ALL The control group included 11 childhood patients with no malignant hematological diseases. Results Aven mRNA expression was found to be higher in patients ≥10 years old and was also significantly higher in relapsed patients. Univariate and multivariate analysis indicated that Aven overexpression was an independent poor prognostic factor. Conclusion Aven mRNA expression is abnormal in childhood ALL and is an independent poor prognostic factor.

BACKGROUND: Clinical observation demonstrates that accelerated fracture healing or lower limb heterotopic ossifications always occur in patients with paraplegia. It indicates that peripheral nervous system may play an important role in fracture healing process.OBJECTIVE: To observe bone histomorphometery parameter, callus formation and biochemical change during the process of fracture healing of unilateral lower limb denervated tibia.DESIGN: Self-control animal experiment.SETTING: Tianjin Hospital.MATERIALS: Totally 36 six-month-old healthy male Wistar rats, with mean body mass of 210 g, were used in this experiment.METHODS: This experiment was carried out at Animal Experimental Center of Tianjin Hospital from March 2001 to March 2004. Denervated tibia fracture model and innervated tibia fracture model were made in the same rat. Animals were executed under anaesthetic status at week 2 and week 4 after fracture. Bilateral tibias were chosen to take radiografts.Biomachamical strength was measured and non-decalcification sections were prepared to perform bone histomorphometery observation.MAIN OUTCOME MEASURES: ① Comparison of wet weight of bilateral tibias and callus of rats between two groups after fracture. ②X-ray plain film scoring. ③ Biomechanical testing of tibial samples. ④ Histomorphological observation of fracture healing RESULTS: ① Wet weight of bilateral tibia and callus of rats in denervated group was much higher than that in innervated group at weeks 2 and 4 after fracture [(0.94±0.15) vs (0.76±0.14) g, (1.06±0.26)vs (0.81±0.10) g,P ＜ 0.05]. ②In X-ray plain film scoring, callus formation was significantly increased in denervated group (P ＜ 0.01). ③In biomechanical testing of three-point bending of tibial sample, callus intensity was significantly lower at weeks 2 and 4 after fracture in denervated group than in innervated group[ (9.88±8.49)vs ( 16.62±13.38 ) N, ( 12.77±7.55 )vs (20.19±10.60) N,P ＜ 0.05]. ④Bone histomorphometery showed that compared with innervated group, mineralized bone trabecula width of denervated group was significantly reduced (P ＜ 0.05), osteoid width was increased , osteoclast index and bone absorption area were significantly increased (P ＜ 0.05), and there were no significant difference of fibroblast index and bone formation area between two groups; Compared with innervated group, mineralized deposition rate in the denervated group was significantly reduced (P ＜ 0.05), the mature time of osteoid was elongated (P ＜ 0.05).CONCLUSION: Peripheral nervous system may play an important role during early and middle period of fracture healing. Intact innervation is essential for normal fracture healing.