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 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 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.

Acute Disease ; Adolescent ; Adult ; Electromyography ; Female ; Humans ; Male ; Middle Aged ; Nervous System Diseases ; chemically induced ; diagnosis ; Organophosphate Poisoning ; Pesticides ; poisoning

Objective To analyze the effect and mechanism of trichostatin A（TSA）on cell cycle in human ovarian cancer cells.Methods Human ovarian cancer cell line A2780 cells were cultured in RPMI 1640 supplement.Flow cytometry analysis and RT-PCR were used to examine the distribution of cell cycles and the level of p21WAF/CIPI mRNA.Results TSA induced increase of G2/M cells increased after the treatment of TSA for 36 hours（P 0.05）;the level of p21WAF/CIPI mRNA expression was upregulated after TSA treatment for 12 hours,the highest leve of its expression occurred at 24 hours,the expression level begun to decrease at 48 hours（P 0.05）.TSA simultaneously induced the decrease of S phase cells in a concentration-dependent manne（rP 0.05）.TSA upregulated the expression of p21WAF/CIPI mRNA in a concentration-dependent manner（P 0.05）.Conclusion TSA could block the G2/M phase and inhibits cell proliferation of A2780 cells through upregulating the expression of p21WAF/CIPI mRNA and the activate cyclin-dependent kinase.

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.

OBJECTIVE Topreparealipidderivativeofgemcitabine(Gem)anditspolymericmi-celles to overcome the disadvantages of Gem.METHODS N-benzyl-3′-acetyl-gemcitabine(BAG)was synthesized.A BAG-loaded poloxamer polymeric micelle (BAG∶poloxamer 188 =10∶1 ,mol/mol)was prepared using an injection method.The micelles were characterized with a laser particle size and elec-tric charge instru ment and negatively-stained trans mission electron microscopy.Hu man breast cancer cells MCF-7 were cultured with Gem or BAG polymeric micelles of 5,10,20,30,50,70,90 μmol·L-1 for 24,48 and 72 h,respectively.The inhibitory rate of cells was measured with an MTT method.The MCF-7 cytotoxicity of BAG polymeric micelles was investigated.A pharmacodynamic study was per-formed on the mice bearing mouse hepatocellular cancer cells H22.Intravenous (iv)and oral (ig)ad-ministration was used at the dose of Ge m 40 mg·kg -1 or BAG polymeric micelles 62 mg·kg -1 .The mice were administered on the 1 st,4th and 7th day and sacrificed on the 8th day.Tumor inhibitory rates were measured.RESULTS TheBAGstructurewasidentifiedbythinlayerchromatograph,1Hand13C NMR,infrared ray chromatograph and mass spectrum.The appearance of BAG micelles was a slightly blue suspension.The micelles were spheres according to the electron microscopic observation.Their size was 62.82 nm and the zeta potential was -18.8 mV.The half inhibition concentration (IC50)of Gem and BAG polymeric micelles was 40.6 and 90.0 μmol·L-1 ,5.0 and 14.9 μmol·L-1 ,5.0 and 1 3.6 μmol·L-1 at 24,48 and 72 h,respectively according to the MTT results.According to the in vivo results,compared with the tumor model group,Gem (ig),Gem (iv)and BAG polymeric micelles (iv and ig)had significant effect on the tumor weight of H22 cell xenograft mice (P<0.01 ).As for anti-tumor efficiency,BAG polymeric micelles (ig)were better than Gem (ig)(P<0.05);BAG polymeric micelles (iv)were better than BAG polymeric micelles (ig)(P<0.05),and BAG polymeric micelles (iv)were almostequaltoGem(iv).CONCLUSION ThelipidderivativeofGemcanbeloadedinthepoloxamer 1 88 polymeric micelles.BAG polymeric micelles show in vitro MCF-7 cell inhibition and in vivo inhibition of mouse H22 xerografts;iv or ig.BAG polymeric micelles (ig)show better anti-tumor effect than Gem (ig),indicating that BAG polymeric micelles are a promising novel anti-tumor oral preparation.

Objective To investigate the correlation between Framingham stroke risk profile(FSRP) and vascular cognitive impairment in stroke-free patients with cerebrovascular risk factors.Methods One hundred and eighty-four stroke-free subjects,selected from Zhejiang hospital,were divided into low risk group (56 subjects),moderate risk group (70 subjects) and high risk group (58 subjects) according to their FSRP score,and their cognitive function including memory ability,attention,executive function and language ability were assessed by Montreal cognitive assessment (MoCA),auditory verbal learning test (AVLT),digit symbol test,trail making test(TMT),digit span and verbal fluency test.Results The total MoCA scores which were (7.2±4.6),(13.8±3.9),(29.6± 12.7) respectively,AVLT-delay recall scores which were(8.2± 1.6),(6.7± 1.4),(5.9± 1.5) respectively,and digit symbol test score which were(34.7±9.3),(32.6± 16.4),(29.7± 13.6) respectively in low,intermediate and high risk groups,decreased with the increasing risk of stroke(P＜0.05).The elapsed time in TMT-B which were (115.2 ±36.9) s,(147.6±44.8) s,(173.9±58.5) s respectively in low,intermediate and high risk groups,prolonged with the increasing risk of stroke (P＜0.05).FSRP was associated with cognitive function,but inversely related to MoCA,AVLT-delay recall,digit symbol test,TMT-B and digit span fall back (P＜0.05),but positively related to consuming time in TMT-B (P＜ 0.05).Multivariate regression analysis showed that advanced age,hypertension,diabetes and smoking were the risk factors for vascular cognitive impairment(P＜0.05).Conclusion Advanced age,smoking,hypertension and diabetes are the most important in vascular risk factors for cognitive impairment.Vascular risk factors can damage cognitive function with the increased risk of stroke,among which delayed recall and executive function are the main affected cognitive area.

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.