Objective To summarize and discuss the clinical results of lumbar catheter drainage auxiliary ventricle drainage surgical puncture on patients with intraventricular hemorrhage about postoperative ADL.Methods The 86 patients with intraventricular hemorrhage admitted to our hospital were randomly divided into two groups,43 patients in control group were given conventional lateral ventricle puncture and continued external drainage,43 patients in observation group were given lumbar catheter drainage auxiliary ventricle drainage surgical puncture based on the conventional treatment.Clinical efficacy were observed,and functional outcomes were evaluated.Results The clearance rate of intraventricular hemorrhage and hospitalized days of the observation group were lower than those of the control group (t =4.23,11.82,all P ＜ 0.05),Quality of life in the observation group was better than that in the control group(x2 =1.73,P ＜ 0.05).Conclusion The clinical result of lumbar catheter drainage auxiliary ventricle drainage surgery for intraventricular hemorrhage in patients is satisfactory and worthy of clinical application.

Objective To study the relationship between the MDR1 gene expressions and CD56 antigen expression in patients with de novo acute myeloid leukemia(AML) and to explore the role of this two factors in clinical drug resistance and their correlation. Methods A real-time quantitative RT-PCR method was established for detecting MDR1 expression levels and three-color flow cytometry analysis using CD34/ SSC gating was used to examined CD56 antigen expression in 79 de novo AML patients. Results CD56 an-tigen was recorded in 19 out of 79 cases (24.1%) and particularly in those with M5 cytotypes. Moreover, CD56 expression was significantly associated with unfavorable cytogenetic abnormalities (P<0.05), Patients with t(8:21)had a significantly higher incidence (57.1%, 4/7) of CD56 expression than those with favora-ble karyotype(P<0.05). CD56~+ AML patients had a higher incidence of splenohepatomegalia and lactate dehydrogenase level than CD56~- patients(P<0.05). The median expression levels of MDR1 was statistical-ly higher in CD56~+ AML patients than that in CD56 patients(P<0.001). Patients with both high levels of MDR1 and CD56~+ had a significantly lower CR(complete remission) rate than those with both low MDR1 level and CD56 (58.8% vs 89.2%, P<0.01). Conclusion There is a linear correlation between MDR1 gene expression and CD56 expression in AML. Quantification of the MDR1 gene expression together with CD56 antigen expression is more effective to the judgement of prognosis in AML.

Objective To analyses the magnetic resonance imaging(MRI)findings of different clinical patterns of neurosyphilis(NS).Methods Clinical records and MRI of 26 patients with NS were retrospectively studied.Results Abnormal MRI was found in 17 patients of 26 patients with NS.In 7 patients were with meningo-vascular syphilis,the MRI commonly showed multiple cerebral ischemia focus and cerebral infarction focus,very few similar to those of encephalitis;Six patients had general paresis,who presented cerebral MRI abnormalities of frontal and temporal atrophy,and few simultaneously with cerebral ischemia focus,granular apendymitis and hippocampus sclerosis;Three patients had syphilitic myelitis,their MRI showed mild tumefaction with multiple ischemic focus all the way through lower cervical spinal cord to lower thoracic spinal cord:One patient was with tabes dorsalis,whose cerebral MRI showed ischemic locus.Another 9 patients had normal MRI,of whom 4 patients with meningitis NS and 5 with tabes dorsalis.Conclusion The MRI of neurosyphilis has diverse presentations,and clinicians should pay much attention to it.

Objective To investigate the alterations of dopamine,norepinephrine,cortisol,C-reactive protein (CRP),heart rate (HR),respiratory rate (RR) and mean artery pressure (MAP) before and after the laser photoeoagulation for retinopathy of prematurity (ROP) with topical anesthesia and to provide the guideline for improving its routine management and interventions.Methods Thirty children with ROP who received ROP laser photocoagulation in Guang zhou Women and Children's Medical Center from May to Dec.2012 were selected.The blood of the 30 cases of infants were collected at 4 time points:before laser therapy,the end of laser therapy,1 hour and 24 hours after laser therapy.The concentrations of dopamine,norepinephrine,cortisol and CRP in plasma were measured at each time point with radio immunoassay,and the values of HR,RR and MAP of infants were recorded as well.Results The levels of dopamine,norepinephrine and cortisol at the end and 1 hour after therapy were higher than those in the quiet state before therapy,and the differences were statistically significant (t =6.39,2.55 ; t =7.74,2.91 ; t =8.87,2.15 ; all P ＜ 0.05) ; the levels of CRP at the end of therapy,1 hour and 24 hours after therapy had no statistical difference in comparison with those in quiet state before therapy (t =0.06,0.89,1.16; all P ＞ 0.05) ; the levels of HR and RR at the end of therapy,1 hour and 24 hours after therapy had statistical difference in comparison with those in the quiet state before therapy (t =4.33,3.84,3.38 ; t =6.81,4.42,2.96 ; all P ＜ 0.05).The level of MAP at the end of therapy had statisti cal difference in comparison with that in the quiet state before therapy (t =6.10,P ＜ 0.001).Conclusions Infantswho experieneed ROP laser photocoagulation had stress response.Clinicians should pay more attention to monitoring HR and RR of preterm infants receiving retinal laser photocoagulation under topical anesthesia and take active intrvventions in order to relieve the stress response.

BACKGROUND: Recombinant adeno-associated virus 2(rAAV-2) has attracted considerable attention due to its nonpathogenic nature in contrast to other viral vectors such as adenoviral and retroviral vectors in gene therapy attempts.OBJECTIVE: To explore rAAV-2 transduction to bone marrow mesenchymalstem cell(BMSC) in vitro and evaluate the possibility of using rAAV-2 as a vector for gene therapy of acute myelogenous leukemia(AML).DESIGN: An open experiment with cells as the observational subjects.SETTING: Department of Hematology, Nanfang Hospital, Southern Medical University.MATERIALS: The experiment was conducted in the Department of Hematology, Nanfang Hospital, Southern Medical University from February to July 2004. We used passages 3 to 5 BMSCs derived from six de novo AML patients and four healthy volunteers in this study.METHODS: BMSC was isolated from 6 to 10 mL of bone marrow aspirates obtained from the iliac crests of the patients who had been diagnosed as having de novo AML and from those of healthy volunteers. The acquired BMSC was infected by rAAV-2 which contained enhanced green fluorescent protein (rAAV-2-eGFP) at different multiplicity of infection(MOI) (MOI = 1 × 102,1 × 103, 1 × 104, 1 × 105, 1 × 106, 1 × 107) . Then we observed through phase contrast fluorescent microscope and flow cytometer to evaluate green fluorescent protein(GFP) expression 10 to 14 days after transduction. GFP expression was observed as the rAAV-2-eGFP transduced BMSC cultured in vitro. We also observed the in vitro gene expression profile of GFP in rAAV-2-eGFP transduced BMSC which was selected by neomycin ( G418). First, we confirmed GFP expression in BMSC through phase contrast fluorescent microscope, then on flow cytometer to detect the percentage of GFP expression.MAIN OUTCOME MEASURES: The efficiency of rAAV-2-eGFP transduction to BMSC. GFP expression was observed through phase contrast fluorescent microscope and flow cytometer at different time points after transduction.rAAV-2-eGFP to BMSC derived from normal volunteers and AML patients had no significant differences. GFP began to express 10 to 14 days after transduction, and the transduction efficiency ranged from 0. 3% to 1.4%. By changing infection condition, we could not make a higher transduction efficiency( P ＞ 0.05) . One round infection of BMSC by rAAV-2-eGFP at a MOI of 1 × 105 was ( 1. 030 ± 0. 034) %, 3 rounds of infection of BMSC by rAAV-2-eGFP at a MOI of 1 × 105 was (1. 140 ±0. 036)%, and coinfected by LipofectAMINE was (1. 380 ± 0. 054)%. However, 293 cell line which was the package cell of rAAV-2 could be efficiently transduced by AML patients transduced by rAAV-2-eGFP at MOI = 1 × 105: The percentage of GFP expression cell gradually decreased from 1.14% at day 12 after transduction to 0. 6% as cell passaged from 2 to 3, and maintained at a level of 0. 5% to 0. 6% later on till 61 days after transduction. After selected by neomycin(G418) 1 month later, rAAV-2-eGFP transduced BMSCs could maintain a long-term GFP expression at a level of 6.0% in vitro without significant decay within 100 days of observation period after transduction.CONCLUSION: The advantages of rAAV-2 mediated gene transduction lie in safety, no immune response to the host, and long-term expression maintained by the target gene. rAAV-2 and BMSC can be used for in vitro gene therapy, and as a systemic gene delivery system, it might be an alternative for systemic gene therapy in the future.

ObjectiveTo investigate the therapeutic effects of the conditioning regimen with or without total body irradiation on allogeneic hematopoietic stem cell transplantation in acute leukemia.Methods We retrospectively evaluated clinical outcomes in 287 allo-HSCT recipients with acute leukemia (ALL- 105,AML-129,and AUL-53) who received myeloablative conditioning regimen with or without total body irradiation from January 2002 to August 2011.Two hundred and six patients obtained complete remission (CR) and 81 non-remission (NR) before transplantation.One hundred and ninety-nine patients received conditioning with total body irradiation (TBI+ Cy group,9 Gy given over 2 days),and 88 patients received busulfan (BuCy group,3.2 mg·kg-1 ·d-1 ),both followed by cyclophosphamide.ResultsThere were no statistically significant differences in hematopoietic reconstitution,regimen-related toxicity (RRT),graft-versus-host disease (GVHD) and relapse between two groups.For patients with AML and AUL,there was no significant difference in the 5-year survival between the two regimens (P＞ 0.05),while for ALL-CR patients,the TBI + Cy regimen had a higher over survival rate (52.0％ vs.31.3％,LogRank=4.249,P＜0.05) and DFS (50.4％ vs.27.8％,LogRank =4.445,P＜0.05) than BuCy.In TBI + Cy group and BuCy group,the proportion of CD19+ B cells at the first month after HSCT was (4.04 ± 1.86)％ and (1.47 ±0.99) ％ (P＜0.05),that of NK cells at 12th month after HSCT was (23.38 ± 12.19) ％ and (13.11± 7.99) ％ (P＜0.05),and that of CD4+ CD45RO+ cells at 9th month after HSCT was (14.63 ±6.17)％ and (9.07 ± 3.12)％ (P＜0.01),respectively.ConclusionUsing TBI-containing regimen was more effective for treating ALL-CR patients than busulphan-containing regimen,but no difference was found for long-term outcomes in patients with AML and AUL between the two regimens.The 9 Gy TBI-based regimens may not affect recipients' thymic function,T-cells reconstitution and immune tolerance,coming out a non-increase of GVHD.

OBJECTIVETo construct the recombinant plasmid containing human microdystrophin cDNA, and study the microdystrophin expression in vivo and in vitro.

METHODSMicrodystrophin cDNA was obtained from recombinant plasmid pBSK-MICRO digested with restrictive endonuclease Not I, the product was inserted into plasmid pVAX1, resulting in pAMICDYS. And then 3T3 cells were transfected with pAMICDYS. Forty-eight hours after transfection, the expression of the microdystrophin was detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry. Finally, TA muscles of mdx mice were injected with the recombinant plasmid pAMICDYS through i.m. and the pathological change of TA was evaluated by histology, and the expression of microdystrophin in mdx TA was detected by immunohistochemical analysis.

RESULTSThe recombinant plasmid containing human microdystrophin cDNA was constructed successfully. The recombinant plasmid was proved to be able to express microdystrophin protein both in vivo and in vitro. Moreover, treatment of the TA of mdx mice with the recombinant plasmid could decrease the number of centrally nucleated myofibers.

CONCLUSIONRecombinant plasmid containing the microdystrophin gene was constructed successfully, and it could express microdystrophin protein both in vivo and in vitro. It provides basis for further study on microdystrophin as a target gene to treat Duchenne muscular dystrophy (DMD) by electrotransfer, i.v, arterial injection and combining with other exogenous gene to enhance microdystrophin expression.

Animals ; Cloning, Molecular ; DNA Restriction Enzymes ; metabolism ; DNA, Complementary ; genetics ; metabolism ; DNA, Recombinant ; genetics ; metabolism ; Dystrophin ; genetics ; Gene Expression ; Genetic Engineering ; Genetic Therapy ; Genetic Vectors ; metabolism ; Humans ; Immunohistochemistry ; Mice ; Muscular Dystrophy, Duchenne ; genetics ; metabolism ; therapy ; NIH 3T3 Cells ; Plasmids ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

Objective:Patients are breathing freely during adjuvant proton pencil beam radiotherapy after breast conserving surgery. Fluctuation of the thorax may affect the position of the end of the proton beam flow, which needs to be precisely evaluated on a millimeter scale.Methods:For 20 patients with breast cancer treated with proton radiotherapy after breast conserving surgery, PET-CT scan was performed approximately 10 min after the end of proton radiotherapy. The images of PET-CT were processed for ROI determination and sampling line (profile) extraction on a Raystation RV workstation to calculate the actual difference between the predicted and real radioactivity from the same spatial location as obtained by PET acquisition R50. Then, the differences in the spatial location between the actual process of proton irradiation and the planned process were obtained. Depth difference values for each pair of sampling lines were presented. Results:For 20 patients with breast cancer with a median follow-up of 22 months (range 12 - 46 months), all patients survived at the last follow-up, and no radiation pneumonitis was observed during the follow-up period. Among the verification results of 21 cases, the depth difference of evenly distributed was (-0.75±1.89) mm in the primary field and (-0.82±2.06) mm in the secondary field; The depth difference of sequential treatment was (1.81±1.87) mm in the primary field and (1.32±1.74) mm in the secondary field; The depth difference of synchronous addition in the primary field was (-1.47±1.44) mm, and the depth difference in the secondary field was (-1.48±2.11) mm.Conclusion:The results of off-line PET-CT in vivo biological verification show that the accuracy of the dose boundary cut-off was within 3 mm in breast cancer patients, which meets the clinical and physician requirement for the precision in breast cancer treatment.