OBJECTIVETo investigate the methylation status of zonula occludens-1 (ZO-1) gene promoter and its clinical significance in children with stage IV non-Hodgkin lymphoma (NHL) and to provide a basis for further etiological study and early diagnosis of this disease.

METHODSFifty-five children with a confirmed diagnosis of stage IV NHL (40 cases of T-NHL and 15 cases of B-NHL) were selected as the case group, and 20 children with diseases other than hematologic malignancies were selected as the control group. Bone marrow samples were collected from these subjects. Methylation-specific PCR (MS-PCR) was applied to evaluate the methylation status of ZO-1 gene promoter, and the integrated optical density (IOD) was determined. RT-PCR was used to measure the mRNA expression of ZO-1.

RESULTSMS-PCR showed that the methylated bands of ZO-1 gene promoter were found in 39 (70.9%) of 55 patients in the case group before treatment, while no ZO-1 gene promoter methylation was detected in the control group. With close tracking of 47 cases in the study group, consisting of 32 cases of T-NHL and 15 cases of B-NHL, the rates of ZO-1 gene promoter methylation prior to treatment were 72% and 67%, respectively, (P>0.572). The cases of T-NHL and B-NHL showed no significant changes in methylation rate in the early and middle phases of chemotherapy (P>0.05), but they showed significant changes in methylation rate in the late phase of chemotherapy (P<0.05). RT-PCR showed that the NHL cases carrying methylated ZO-1 gene had no mRNA expression of ZO-1, while all children in the control group had mRNA expression of ZO-1. There was no linear relationship between the total number of peripheral blood leukocytes and ZO-1 gene IOD (r=0.093, P=0.575); a positive correlation was found between the number of malignant cells in bone marrow and ZO-1 gene IOD (r=0.669, P<0.001).

CONCLUSIONSZO-1 gene shows a hypermethylation status in children with NHL, and the methylation level is positively correlated with the number of malignant cells in bone marrow. ZO-1 may be used as a novel molecular marker in early diagnosis, outcome assessment, prognostic evaluation, and detection of minimal residual disease.

Adolescent ; Child ; Child, Preschool ; DNA Methylation ; Female ; Humans ; Infant ; Lymphoma, Non-Hodgkin ; genetics ; Male ; Promoter Regions, Genetic ; Zonula Occludens-1 Protein ; genetics

China ; Congresses as Topic ; Fatty Liver ; Humans

Objective To discuss imaging classification of blunt traumatic thoracic aortic injury (BTTAI) and weigh guideline value of its imaging classification to surgical options.Methods BTTAI in 12 patients were divided into three types in accordance with outline of aortic injury revealed by CT imaging,i.e.,type A of normal outline of aortic blood vessel but free endothelium in aortic cavity,type B of abnormal aortic outline and contrast extravasation to aortic lumen exterior but only confining to mediastina,and type C of abnormal aortic outline and contrast leakage to thoracic cavity.Significance of BTTAI morphological classification was analyzed according to data,such as systemic injury severity score (ISS),local injury sites,surgical approaches and patients' outcome.Results Multiple injuries combined with BTTAI were observed in all patients whose aortic lesion image could be all generalized by above-mentioned classification.BTTAI image showed type A in three patients,type B in seven and type C in two.Pseudoaneurysm was confirmed as the foremost common BTTAI.There were no significant differences in ISS,hypotension morbidity,treatment methods or mortality between each type of BTTAI.Conclusions BTTAI occurs mostly in multiple injuries and thus overall injury severity assessment is not only depended on aortic lesion classification.BTTAI classification in this study reflects injury severity of vessel wall,takes account to location of lesion and adjacent relations (especially length of landing zone) and hence has accurate referential value for surgical decisions.

Objective To investigate feasibility and validity of endovascular aortic repair (EVAR) of traumatic thoracic aortic injuries (TTAI).Methods A retrospective analysis was conducted on data of 13 patients with TTAI.Pathological changes were evaluated by spinal CT angiography (CTA) preoperatively and re-evaluated by digital subtraction angiography (DSA) in EVAR.CTA was performed again to confirm therapeutic effects at postoperative 3,6,12 months and annually thereafter.Results All patients had successful EVAR.Complete or partial cover of left subclavian artery was observed in four patients.Endoleak in angiography shortly after stent delivery was noticed in three patients.However,endoleak disappeared in one patient after short stent placement for twice; endoleak was evidently decreased in one patient after balloon dilation.Follow-up was performed for another patient with slight endoleak.A total of 12 patients were followed up,which showed no complications,such as endoleak,ischemia of left upper extremity,paralysis or stent-graft migration.Conclusion EVR is safe and effective in treatment of TTAI.

Animals ; Apoptosis ; Herpes Simplex ; physiopathology ; virology ; Humans ; MicroRNAs ; genetics ; metabolism ; Simplexvirus ; genetics ; physiology ; Virus Latency

Objective To develop mycobacterial inducible expression vectors which permit to overexpress Mycobacterium tuberculosis (Mtb) immunodominant antigen, and to analyze its immunogenicity after purification by affinity chromatography. Methods The regulatory region of M. smegmatis (Ms) acet-amidase(pACE) was obtained by PCR amplification, and was used as promoter to construct the mycobacteri-al inducible expression vectors, pMF series. The coding gene of Mtb chimeric antigen Ag856A2 which is a recombinant Ag85A with 2 copies of ESAT-6 inserted in its Acc Ⅰ site and showed excellent immunogenicity in the animal experiments we described previously, was cloned into the pMF vector series, and was induced to express by the addition of acetamide. The recombinant protein expressed in the Ms was purified by the Ni~(2+)-NTA affinity chromatography, the resulted homologous recombinant antigen was added into the spleen cells separated from BCG vaccinated mice, and the immunogenicity was analyzed by the IFN-γ ELISPOT as-say. Results The mycobacterial inducible expression vectors, pMF series was constructed successfully, target antigen could be. induced to express in the Ms by the addition of 0.02% acetamide, and could be puri-fied by the Ni~(2+) -NTA affinity chromatography due to the addition of 6×His tag in the vector pMF406. Fur-thermore, the mycobactefial homologous antigen could induce more IFN-γ secretion than the heterogonous one. Conclusion The mycobacterial inducible expression system based on the regulatory region of Ms acet-amidase as promoter could permit the Mtb target antigen of interest overexpression and purification, and the immunogenicity of the homologous antigen from Ms is better than that of be expressed from E. coli, which may be more potential for immunological detection of tuberculosis.

Objective To evaluate the dosimetric errors of organs-at-risk (OARs) induced by the optimal auto-segmentation using Mim Maestro based on dose calculation and measurement.Methods The Mim atlas library composed of 240 nasopharyngeal carcinoma,breast cancer,and rectal cancer patients that were retrospectively selected was used for the auto-segmentation of OARs on the CT images of corresponding regions in 76 patients.Relative to the manual contouring,one optimal case was selected from each site based on conformity index (CI),mean distance to conformity (MDC),relative volume difference (Dv％),DICE,sensitivity index (Se.Idx),and inclusion index (Inc.Idx).Treatment plans were made to satisfy the DVH constraints of OARs based on auto-contours,and then the dose errors to the actual organs were evaluated in terms of calculation and measurement.The paired t-test (normal distribution) or rank sum test (non-normal distribution).Results Significant differences were observed in the 76 patients between the manual and automated segmentation (P＜0.05).For the optimal cases,the DICE index of various OARs ranged from 0.43 to O.98,and 73％(16/22) of DICE values were higher than 0.70.The calculated dose errors to various OARs were (-1.15±15.94)％(95％ CI:-8.21％ to 5.92％) (mean dose) and (-6.53±21.13)％ (95％ CI:-15.90％ to 2.84％) (maximum dose).The measured dose errors were (-2.43± 24.52)％ (95％ CI:-13.30％ to 8.44％)(mean dose) and (-3.38±20.87)％(95％ CI:-12.63％ to 5.87％)(maximum dose).Conclusion Without human interference,even the optimal auto-segmentation results are not clinically acceptable for treatment planning.

AIM:To study the feasibility of recombinant adeno-associated virus(rAAV)as a vector to transfer the green fluorescent protein(GFP)gene as a target gene into rabbit retina.METHODS:Intravitreal injection of rAAV-gfp was performed in either eye for each rabbit with the other eye taken as control.At the 3rd,7th,and 14th day after injection,the eyeballs were removed,and the retinas were flat-mounted on glass slides to inspect the retinal fluorescence,respectively.RESULTS:After intravitreal injection of rAAV-gfp,the presence of fluorescent spots in the cytoplasm of retinal cells indicated that GFP gene was efficiently transferred and expressed in the rabbit retina.CONCLUSION:Recombinant adeno-associated virus is a reliable and simple vector for transferring target gene,e.g.,GFP gene,to the retina.

OBJECTIVETo investigate the effect of salidroside on hypoxia-induced apoptosis of corpus cavernosum smooth muscle cells (CCSMCs) in rats.

METHODSRat CCSMCs were cultured in vitro by the enzyme digestion method and identified by immunofluorescent staining of anti-alpha-SMA and anti-Desmin. The non-toxic dose of salidroside was determined by MTT assay. Low-oxygen mixed gas (1% O2, 5% CO2, and 94% N2) was piped into a modular incubator chamber to induce hypoxia. The CCSMCs were divided into a normal, a hypoxia, and a 32 microg/mL salidroside intervention group. The apoptosis of the CCSMCs was detected by flow cytometry and the expression of the caspase-3 protein determined by Western blot.

RESULTSThe majority of the CCSMCs were positive for alpha-SMA and Desmin at immunofluorescent staining. Salidroside at < 32 microg/ml produced no obvious toxicity to CCSMCs. Compared with the normal control group, the rates of early and late apoptosis of CCSMCs were both increased significantly in the hypoxia group ([12.77 +/-1.41]% vs [18.69 +/- 1.29]%, P < 0.01 and [14.63 +/- 2.00]% vs [21.03 +/- 1.530]% , P < 0.05). Western blot showed a markedly increased expression of cleaved caspase-3 (P < 0.01). Intervention with 32 microg/ml salidroside significantly reduced hypoxia-induced early apoptosis of CCSMCs ([13.46% +/- 1.87]%, P < 0.01) and decreased the expression of cleaved caspase-3 (P < 0.01).

CONCLUSIONSalidroside can reduce the expression of cleaved caspase-3 and inhibit hypoxia-induced apoptosis of CCSMCs in rats.

Animals ; Apoptosis ; drug effects ; physiology ; Caspase 3 ; metabolism ; Cell Hypoxia ; physiology ; Cells, Cultured ; Glucosides ; pharmacology ; Humans ; Male ; Myocytes, Smooth Muscle ; cytology ; drug effects ; enzymology ; Penis ; cytology ; drug effects ; Phenols ; pharmacology ; Rats

To study the equipment modular allocation in the emergency medical unit after Beijing emergency public affairs. Using an improved Delphi method, 25 experts from various fields in Beijing were involved into two rounds consultations. The questionnaire of the first round consultation was formed based on analysis of emergency rescue equipment. Based on the result of the first round consultation, the questionnaire of the second round consultation was set up. The two rounds of questionnaires had the responding rates of experts 100% respectively, and the au-thoritative coefficients of the experts were 0.81 and 0.88. An emergency medical rescue equipment solution with three de-grees modular configuration in Beijing was made based on the consultations and experts' advices. The solu-tion develops the emergency medical service ability of mobile medical units and vehicle-mounted mobile hospital. LUO Ying and GAO Hong-wei are the first authors who contributecl equally to the article.