Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction

Objective To study in vitro the inhibitory effects and mechanisms of N-butanol extract of Potentilla anserine L.(NP)against hypoxia-induced nitric oxide(NO)in hippocampus neuron of rats. Methods The models of hippocampus neurons hypoxia injury of Sprague-Dawley(SD)neonatal rats were cultured in vitro. The cultured hippocampus neurons were divided randomly into blank control group, hypoxia injury model group, nimodipine group(2 μmol/L)and NP high(250.0 mg/L),middle(62.5 mg/L),low(15.6 mg/L)dose groups. The activities of hippocampus neurons were examined by methyl thiazolyl tetrazolium(MTT)assay,and meanwhile their contents of nitrogen monoxidum(NO)were detected. Half quantity reverse transcription-polymerase chain reaction(RT-PCR)and Western blotting were used to detect neuronal nitric oxide synthetase(nNOS)mRNA and protein expression levels respectively in each group,immunocytochemistry stain was used to detect protein positive rate. Results Compared with blank control group,the activity of neuron〔absorbance(A)value〕was significantly decreased(0.0826±0.0095 vs. 0.3315±0.0105),content of NO(μmol/g:0.0509±0.0027 vs. 0.0291±0.0032), the expression levels of nNOS mRNA (0.1463±0.0081 vs. 0.0801±0.0058), the positive rate of nNOS〔(74.4238±3.9423)%vs.(28.3714±4.1361)%〕,the expression levels of nNOS protein(A value:1.9130±0.0471 vs. 0.5068±0.0368)were all significantly increased in the hypoxia injury model group(all P<0.01). Compared with hypoxia injury model,the activity of neuron was increased,contents of NO,the expression levels of nNOS mRNA,the positive rate of nNOS,the express levels of nNOS protein were decreased in each medicine group,especially prominent in the NP high concentration group〔the activity of neuron:0.1681±0.0118,contents of NO:0.0319±0.0044,nNOS mRNA:0.0648±0.0032,nNOS positive rate:(40.1240±6.4900)%,nNOS protein:1.3924±0.0621,all P<0.01〕. There were no statistical significant differences between the NP low concentration group and model group(all P>0.05). Conclusions NP can ameliorate the injury of rat hippocampus neurons induced by hypoxia in vitro. The possible mechanisms might be related to the effective inhibition of the synthesis of nNOS and NO excessive generation.

Objective To improve the recognition of treatment of adult T lymphoblastic lymphoma (TLL) with Hyper-CVAD regime.Methods The turnovers of 7 cases treated with Hyper-CVAD regime were summarized.Results Among the 7 cases,1 case died after the first unfinished chemotherapy,1 case gained complete remission before autologous stem cell transplantation but relapsed after transplantation,3 cases gained complete remission until now,2 cases without transplantation relapsed and then gave up following treatments after treating with ICE regime or the virgin regime,but the therapeutic effects were poor.Conclusions Adult TLL should get early chemotherapy treatments,and some special characteristics should be noted,the treatment does not relate to the stage,the chemotherapy treatment should be carried out regularly and enough,radiation of mediastinum mass can be applied after 4 cycles of Hyper-CVAD regime,choose suitable transplantation method.

Objective To explore the curative effect of ursodeoxycholi acid on retinopathy of prematurity (ROP) caused by high concentration of oxygen in newborn rats. Methods The model of ROP was established. Neonatal rats were divided into normal control group, ROP model group, low dose ursodeoxycholi acid treatment group (10mg/kg) and high dose ursodeoxycholi acid treatment group (40mg/kg). Rats were sacrificed at days 17. The new retinal vessels were observed and counted under lfuorescence microscope. Results The new retinal vessels in ROP rats were hyperplastic, twisted and unevenly distributed. There was signiifcant difference in the number of new retinal vessels among different groups (P=0.000). The number of new retinal vessels of rats in ROP group, low-dose group and high-dose group was signiifcantly more than that in control group (P=0.000). The number of new retinal vessels in low-dose group and high-dose group was significantly less than that in ROP group (P<0.05). The number of new retinal vessels in high-dose group was less than that in low-dose group (P>0.05). Conclusions Ursodeoxycholi acid could inhibit the angiogenesis of retina and could have curative effect on ROP.

BACKGROUND:Conventional methods,including trypsin digestion and cells transfer using single call suspension,have many drawbacks,which limit the development of bone tissue engineering.OBJECTIVE:To culture bone marrow stromal stem calls,induce osteoblastic differentiation,and prepare cell sheets.METHODS:Canine bone marrow stromal calls were isolated by density gradient centrifugation technique,inoculated into DMEM medium,and induced to differentiate into osteoblasts.Complete call sheets were harvested by call sheet engineering based on the temperature change of temperature-responsive medium.RESULTS AND CONCLUSION:Immediately after inoculation,primary calls were scattered on the bottom of culture flask,presenting a transparent spherical body with a good refractive capacity.At 12 hours,calls exhibited a long shuttle shape,reached complete confluency,and grew in a whirlpool-like fashion.After osteoblastic induction,the majority of bone marrow stromal stem calls appeared tetragonal,polygonal,and squamose.At 21-28 days,round or oval-shaped calcified nodules formed.When the bone marrow stromal stem calls in the temperature-responsive culture dishes were cooled below the critical temperature 32℃,cells were gradually detached from the bottom of culture flask and formed complete bone marrow stromal stem call sheets.These findings indicate that density gradient cantrifugation technique can be used to successfully isolate and culture canine bone marrow stromal stem cells to differentiate into osteoblasts and call sheet engineering enables to harvest complete bone marrow stromal stem call sheets.

Objective To analyze the risk factors associated with allergic disease in preterm infants during the first year of life. To investigate whether there was any association between peripheral blood eosinophils and allergic di-sease during the first year of life benefit of hydrolyzed formula feeding in preterm infants. Methods A nested case -control study was conducted in a cohort of artificial feeding preterm infants admitted to Neonatal Intensive Care Unit, Shanghai Children's Medical Center from January 2012 to April 2013. The preterm infants were monitored from birth up to 1 year of age and the findings were related to development of allergic disease. Individuals with allergic disease during the first year of life(cases)were matched with gender,as previously described with two individuals(controls)who re-mained event - free during the study. The risk factors associated with allergic disease in preterm infants during the first year of life were analyzed. And whether there was any association between peripheral blood eosinophils and allergic di-sease during the first year of life benefit of a hydrolyzed formula feeding in preterm infants was investigated. Different variables in control and case individuals were compared with t test for normal distribution measurement date,χ2 tests for categorical variables and Mann - Whitney U test for non - normal distribution measurement date,and multiple conditio-nal Logistic regression were used to investigate the risk factors associated with allergic disease in preterm infants. Results Thirty - four individuals were in cases and 68 individuals were in controls. In a conditional multivariable Logistic model,peripheral blood eosinophils of preterm infants at full enteral feeding(EOS - 2)(OR = 5. 941,95% CI:1. 165 - 41. 375,P ﹤ 0. 05),and family history of allergy(OR = 3. 316,95% CI:1. 201 - 9. 152,P ﹤ 0. 05)were the two independent risk factors for allergic disease in preterm infants during the first year of life. Individuals fed with standard preterm formula after birth,the association between EOS - 2 and allergic disease was significantly enhanced (OR = 21. 459,95% CI:1. 686 - 273. 152,P ﹤ 0. 05). By contrast,in individuals fed with hydrolyzed formula,the risk of EOS - 2 was substantially attenuated(OR = 1. 708,95% CI:0. 148 - 19. 743,P ﹥ 0. 05). Conclusions Peripheral blood eosinophils of preterm infants at full enteral feeding EOS - 2 and family history of allergy were the 2 independent risk factors for allergic disease in preterm infants during the first year of life. Contrast to individuals fed with standard preterm formula after birth,individuals fed with hydrolyzed formula had lower association between factors of peripheral blood eosinophils and family history of allergy,and allergic disease during the first year of life.

Objective To explore the impacts of enriched environment(EE),which has different initiation time points and intensity,on the neural and ethological prognosis and contents of myelin basic protein(MBP)of neo-natal rats with hypoxic - ischemic brain damage(HIBD). Methods HIBD rat models were established. Rats were divided into the early,the intermediate and the late intervention groups,which experienced EE from 7,14 and 21 days after HIBD for 14 days. The early and intermediate intervention groups were then divided into 6 - h and 24 - h groups, which experienced EE intervention for 6 hours or 24 hours respectively each day. Trapeze tests and water maze tests were carried out to detect the neural and ethological prognosis. Immunohistochemical method was used to detect MBP of the brain white matter,and the percentages of positive cells with MBP were detected by an image analyzer. The con-tents of MBP were measured. Results The trapeze test scores of the early and intermediate sham operation group,HI group,early 6 - h and 24 - h EE groups and the intermediate 6 - h and 24 - h EE groups,the late sham operation group,the late HI and late EE intervention group were(4. 05 ± 0. 88)scores,(2. 35 ± 1. 02)scores,(3. 67 ± 1. 12) scores,(3. 50 ± 1. 41)scores,(3. 50 ± 0. 93)scores,(3. 56 ± 1. 13)scores,(4. 00 ± 0. 89)scores,(2. 17 ± 1. 17)scores,(3. 50 ± 0. 92)scores,respectively. The trapeze test scores of early,intermediate and late EE groups were higher than those of the HI groups in the same period. There was no significant difference between the early,the intermediate 6 - h EE groups and 24 - h EE groups. Scores of water maze of each corresponding group were(40. 68 ± 23. 77)seconds,(56. 66 ± 10. 96)seconds,(46. 49 ± 19. 27)seconds,(51. 72 ± 20. 46)seconds,(38. 20 ± 18. 36)seconds,(47. 96 ± 20. 65)seconds,(38. 63 ± 20. 44)seconds,(59. 66 ± 13. 81)seconds and(45. 93 ± 22. 45)seconds,respectively. The water maze scores of the early,the intermediate 6 - h EE group and the late 24 -h EE groups were higher than those of the HI groups in the same period. There was no significant difference between the early,the intermediate 6 - h EE groups and the 24 - h EE groups. The relative abundance of MBP of the early and intermediate and the late HI groups were 6. 32 ± 1. 63 and 6. 74 ± 2. 19,and significantly less than that of the sham groups in the same periods,which were 9. 09 ± 1. 69 and 9. 37 ± 2. 46. The relative abundance of MBP of early 6 - h and 24 - h groups,the intermediate 6 - h and 24 - h groups and the late EE group was 7. 84 ± 2. 51,8. 05 ± 1. 86, 8. 89 ± 2. 29,8. 48 ± 2. 67 and 7. 98 ± 2. 09,respectively,which was significantly higher than that of the HI groups in the same periods. It showed that the neural and ethological prognosis of neonatal rats with HIBD could be improved,no matter the intervention began in the early,the intermediate or the late periods,or the intervention time was 6 hours or 24 hours each day. And relative abundance of MBP in the white matter increased with EE. Conclusions EE interven-tion has a long window stage for young rats. EE intervention could improve the neural and ethological prognosis of rats with HIBD. EE intervention could elevate the contents of MBP in the white matter,which could be one of the mecha-nisms for EE to improve the neural and ethological prognosis of rats with HIBD.

Objective To investigate the effects of Saussurea involucrata extract pretreatment on the expression of the Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) in focal cerebral ischemia/reperfusion in mice and its possible neuroprotective mechanism.Methods Seventy-two Kunming mice were randomly divided into four groups:sham operation,saline,Saussurea involucrata extract,and edaravone groups (n =18 in each group).Saussurea involucrata extract 0.8 g/kg was given intraperitoneally in the Saussurea involucrata extract group; edaravone 3 mg/kg was given in the edaravone group; and the same volume of saline was given in the saline group.A model of middle cerebral artery occlusion (MCAO) was induced after 7 days of continuous injection.Cerebral infarct volume was determined by 2,3,5-triphenyltetrazolium staining.Immunohistochemical staining was used to detect TLR4-positive cells in ischemic brain tissue.Reverse transcriptase polymerase chain reaction was used to detect the expression of TLR4/NF-κB mRNA.Results The cerebral infarct volume in mice in the saline,Saussurea involucrata extract and edaravone groups was 131.55± 28.25 mm3,84.10 ±13.92 mm3 and 65.10 ± 6.78 mm3,respectively.There were significant difference (F =10.158,P =0.012).The infarct volume in the Saussurea involucrata extract group (P =0.020) and edaravone group (P0.005) was significantly less than that in the saline group,and there was no significantly difference between the 2 groups.The numbers of cortex and TLR4 positive cells in hippocampus area at the ischemic sides in the saline group were significantly more than those in the sham operation group (all P ＜0.001).The numbers of positive cells of cortex and TLR4 in the Saussurea involucrata extract group and the edaravone group were significantly decreased compared to the saline group (all P ＜ 0.05),and there was no significant differences between the Saussurea involucrata extract group and the edaravone group.The expressions of TLR4,p50,and p65 mRNA in the saline group were significantly up-regulated compared to the sham operation group (all P =0.000).Saussurea involucrata extract could significantly down-regulate the expressions of TLR4,p50,and p65 mRNA at 24 hours after ischemia/reperfusion (all P =0.000).Edaravone could significantly down-regulate the expressions of TLR4 and p65 mRNA (all P =0.000) and it had a down-regulated trend for the expression of p50 mRNA (P =0.053); while there was no significant difference in the expressions of TLR4 and p65 mRNA between the Saussurea involucrata extract group and the edaravone group.Conclusions Saussurea involucrata extract pretreatment may significantly reduce the cerebral infarct volume,down-regulate the expressions of TLR4 and NF-κB subunit,and play a neuroprotective effect by inhibiting inflammatory response after ischemia.

Objective Salidroside is a major active component of integripetal rhodiola herbal medicine, which has a significant activity against hypoxia and ischemia.This study was to investigate the effects of side chain carbon losssalidroside analogues (N04) on the expressions of HIF-1α-related factors in the hypoxia-injured EAhy926 human vascular endothelial cells.Methods EAhy926 human umbilical vein endothelial cells in the logarithmic growth phase were randomly divided into a normal control, a hypoxia model control, a salidroside, a high-dose N04, a medium-dose N04, and a low-dose N04 group.The hypoxia model was established by depriving the culture medium of sugar and serum and culturing the EAhy926 cells in an environment of 95%N2+5%CO2 for 2 hours, followed by intervention with salidroside at 1×10-6 mol/L and N04 at 1×10-6, 1×10-7, and 1×10-8 mol/L, respectively.Then, the activity of the cells was detected by MTT assay, their LDH activity examined by spectrophotometry, the mRNA expressions of HIF-1α and VEGF measured by RT-PCR, the protein expressions of HIF-1α, VEGF and pVHL determined by Western blot, and the activity of eNOS measured by ELISA.Results Compared with the normal control group, the hypoxia model cells showed significantly reduced activity (0.51±0.05 vs 0.27±0.02, P<0.01), an elevated LDH level ([6.65±1.43] vs [78.82±2.33] U/L, P<0.01), and decreased eNOS activity ([1.56±0.23] vs [1.16±0.20] U/100 mL, P<0.01).In comparison with the hypoxia model group, the cells treated with high-, medium-, and low-dose N04 exhibited remarkably increased activity (0.27±0.02 vs 0.0.42±0.05, 0.40±0.03 and 0.37±0.04, P<0.01), a reduced LDH level ([78.82±2.33] vs [53.05±3.90], [58.42±4.45] and [62.73±3.63] U/L, P<0.01), and increased eNOS activity ([1.16±0.20] vs [3.01±0.47], [2.60±0.26] and [2.32±0.29] U/100 mL, P<0.01).The activity of eNOS was also increased in the salidroside group ([2.32±0.29] U/100 mL, P<0.01).The cell activity in the high-and medium-dose N04 groups was markedly higher than that in the salidroside group (P<0.05), and so was the eNOS activity in the high-dose N04 group and the LDH level in the medium-and low-dose N04 groups (P<0.05).In comparison with the normal control group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were significantly up-regulated in the hypoxia model group (P<0.01) while that of the pVHL protein markedly down-regulated (P<0.01).Compared with the hypoxia model group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were remarkably reduced (P<0.05), while that of the pVHL protein markedly elevated (P<0.05).Both the expressions of VEGF mRNA and HIF-1α protein were significantly lower in the medium-and low-dose N04 groups than in the salidroside group (P<0.05).Conclusion N04 can protect vascular endothelial cells against hypoxia-induced injury by regulating the expression of HIF-1α-related factors and eNOS.

Objective To investigate the safety and efficacy of endoscopic submucosal dissection (ESD) in elderly patients (age ≥ 65 years) with early gastric cancer.Methods The retrospective cohort study was adopted.The clinical data of 145 patients with early gastric cancer who underwent ESD at the Peking University Cancer Hospital between January 2011 and June 2014 were collected.Of 145 patients,57 (age≥65 years) were allocated into the elderly group and 88 (age ＜ 65 years) were allocated into the non-elderly group.ESD was performed to all the patients after the multidisciplinary discussion and evaluation.Observation indicators included operation time,volume of intraoperative blood loss,duration of hospital stay,complications,integrity of resected specimens,negative resection margin,tumor cure and follow-up (survival,tumor recurrence and metastasis).The follow-up using outpatient examination and telephone interview was performed to detect survival of patients and tumor recurrence and metastasis till June 2015.Measurement data with normal distribution were presented as x ± s and comparison between groups was analyzed using t test.Measurement data with skewed distribution were presented as M (range) and comparison between groups was analyzed using nonparametric U test.Count data were analyzed using the chi-square test.Results Operation time,volume of intraoperative blood loss,duration of hospital stay,numbers of patients with postoperative perforation,bleeding,en bloc resection of tumor,piecemeal resection of tumor,negative resection margins,positive resection margins,curative resection,extended curative resection and non-curative resection were 100 minutes (range,20-470 minutes),25 mL (range,5-200 mL),5 days (range,2-10 days),1,2,52,5,50,7,30,17,10 in the elderly group and 110 minutes (range,25-480 minutes),25 mL (range,5-600 mL),4 days (range,2-29 days),3,3,85,3,83,5,47,24,17 in the non-elderly group,respectively,with no significant differences between the 2 groups (U =2 451.000,2 183.500,2 116.500,x2=1.544,1.018,1.210,0.142,P ＞ 0.05).Patients with complications were improved after symptomatic treatment.Of 145 patients,135 were followed up for a median time of 26 months (range,12-53 months).Tumor recurrence rates in the elderly and non-elderly groups were 1.8％ (1/57) and 2.3％ (2/88),showing no significant difference between the 2 groups (x2=0.000,P ＞ 0.05).No tumor recurrence and metastasis and death were occurred in other patients.Conclusion ESD is safe and feasible in the treatment of elderly patients with early gastric cancer,and it is equivalent to short-and long-term efficacies of ESD in non-elderly patients.