Objective To explore cause of death and the lessons learned in critically ill patients with organophosphate poisoning.Methods The clinical manifestations,treatment,cause of death in 31 patients with severe organophosphate poisoning deaths were retrospectively analyzed.Results 31 deaths of patients,12 patients died of cholinergic crisis(38.70％),16 patients died of respiratory failure(51.61％),2 patients died of atropine intoxication (6.45％),1 patient died of the solution be excessive phosphorus(3.23％).6.01％ incidence of intermediate syndrome,mortality was 16.94％.Conclusion Poison not completely clear,atropine and cholinesterase agents applied inappropriately,patients with mechanical ventilation for respiratory failure improper treatment,the merger is an important organ of the original basis of disease leading cause of death,accurately determine the condition is the key to reducing mortality.

BACKGROUND: Mesenchymal stem cells (MSCs) are commonly observed under the scanning electron microscope, transmission electron microscope and inverted microscope. However, above-mentioned observation methods have disadvantages on observing ultrastructure of cell membrane and cytoskeleton. OBJECTIVE: To establish a more effective and appropriate method to isolate, culture and identification of human umbilical cord mesenchymal stem cells (hUCMSCs) in vitro and to study the ultrastructure of osteogenic differentiation with the atomic force microscope. METHODS: The hUCMSCs were isolated from human umbilical cord by digested with collagenase. After serial subcultivation in vitro, the stem cells were passaged, and osteogenic differentiation was determined by alkaline phosphatase calcium-cobalt staining and alizarin red staining. hUCMSCs immunophenotype was measured by Flow cytometry.The membrane surface ultrastructure of osteogenic differentiation was observed by Atomic Force Microscope before and after induction. RESULTS AND CONCLUSION: The isolated hUCMSCs by digested with collagenase was efficient. After seeded 24 hours, the adherent cell showed spindle shape, polygonal shape and fibroblast-cell-like shape and the size of hUCMSCs was homogeneous. Flow cytometry analysis revealed that CD29, CD44, CD105 were highly expressed on the surface of passages 3 cells, but there was negative for CD34, CD45 and HLA-DR. These cells were high positive for alkaline phosphate staining and also showed significant calcium node using alizarin red staining after 4 weeks culture induction of osteogenic differentiation. Under atomic force microscopy, undifferentiated stem cells demonstrated insignificant microtubule protrution in a parallel distribution following analysis of cytoskeleton before and after differentiation.

Crohn's disease (CD) is a chronic nonspecific inflammatory disease.CD can affect any location in the digestive tract,and it also affect other organs,including the eyes,skin,liver and joints,which are termed extraintestinal manifestations (EIMs).The cutaneous manifestations of CD are common and occur in about one-third of patients.EIMs of CD have been divided into 3 categories.(1) Specific lesion,cutaneous manifestations of CD were the same as histopathologic findings of underlying gastrointestinal lesion.(2) Reactive lesion,it was also inflammatory lesion which was usually accompanied by underlying gastrointestinal disease while inflammatory injury was different from histopathologic findings of gastrointestinal lesion.(3) Associated lesion,it was caused by sequelae of human leucocyte antigen and chronic inflammation.In the current era of ever-expanding therapeutic options for CD,some investigators have proposed a fourth category of EIMs,namely those that are therapy-related lesion.The therapy-related lesion is closely related to disease-associated conditions in light of certain skin findings,and there is potential overlap between them.

Objective:To investigate the efficacy and safety of less invasive surfactant administration(LISA) of neonatal respiratory distress syndrome(NRDS).Methods:From July 2017 to December 2018, 50 premature infants with birth weight ≤1 500 g and/or gestational age≤32 weeks diagnosed as NRDS at the Fifth Medical Center of PLA General Hospital were randomly divided into LISA group( n=25)and INSURE group( n=25). The patients in LISA group was inserted fine duct into the trachea through direct laryngoscope under nasal continuous positive airway pressure (nCPAP) and pulmonary surfactant was injected.The INSURE group adopts endotracheal intubation-pulmonary surfactant-nCPAP was performed after unplugging.The changes of vital signs, blood gas indexes, adverse reactions and the incidence of complications were compared between two groups at different time points. Results:There was no significant difference in respiration, heart rate, systolic blood pressure, diastolic blood pressure and PaO 2, PaCO 2, BE, SpO 2 between two groups at different administration time points.Although the pH value of LISA group was lower than that of INSURE group, it was within the normal range.There was no significant difference in bronchopulmonary dysplasia, intraventricular hemorrhage, periventricular leucomalacia and other complications between two groups, and there was no death, air leakage, retinopathy of prematurity and pulmonary hemorrhage in both two groups.In addition, there was no significant difference in hospitalization days, total medical expenses, oxygen use time between the two groups(all P>0.05). Conclusion:Compared with INSURE technology, LISA technology has its feasibility for premature infants with NRDS, but the effectiveness and safety in the practical application need to be further confirmed.

Obje:ctive To establish an optimized method to isolate, culture and identify human umbilical cord mesenchymal stem cells (hUCMSCs) in vitro and induce their osteogenic and adipogenic differentiation. Methods The hUCMSCs were isolated from human umbilical cord by digestion with collagenase. After serial subcultivation in vitro, the stem cells were passaged. Morphologic appearance of hUCMSCs was observed under an optical microscope and atomic force microscope. The proliferation rate was measured by MTT assay. Cell cycle and surface antigens were measured by flow cytometry. The osteogenic and adipogenic differentiation was tested and evaluated by specific staining methods. Results The isolation of hUCMSCs by digestion with collagenase was efficient. After seeded for 24 hours, the adherent cells showed spindle shape and fibroblast cell-like shape and the size of hUCMSCs was homogeneous. The similar growth curves of passage 3 and 7 exhibited a great potential for proliferation. Flow cytometry analysis revealed that CD29, CD44 and CD105 were highly expressed on the surface of passages 3 cells, but the expression was negative for CD34, CD45 and HLA-DR. After culture in inducing medium, the cells were successfully induced into osteogenic and adipogenic lineages. These cells were highly positive for alkaline phosphate staining and also showed mineralization presented with von kossa staining after 4 weeks' culture induction of osteogenic differentiation. Furthermore, liquid vacuoles were detected by oil red O staining after 3 weeks' culture induction of adipogenic differentiation. Conclusion An in vitro method for isolation and purification of hUCMSCs from human umbilical cord has been established. The cultured cells were composed of only undifferentiated cells and their biological properties were stable. The hUCMSCs are expected to be a new type of stem cells of tissue engineering.

Objective To investigate the expression of miRNA-16 in peripheral blood monouclear cells (PBMC)from systemic lupus erythematosus( SLE)patients. Methods Sixteen SLE patients who meet the diagnostic criteria of SLE revised in 1997 American rheumatology and 12 healthy individuals were selected as our subjects. Their peripheral blood were sampled. Total RNAs were extracted and purified. The level of miRNA-16 was determined by quantitative reverse transcription PCR( qRT-PCR). U6 was used as housekeeping control. The amount of target miRNA was normalized relative to the amount of U6(ΔCt =ΔCt miRNA-ΔCtU6 ). Relative expression levels were expressed as 2-ΔCt . Results The expression level of miRNA-16 in the SLE patients was 919. 87 ± 715. 45,significantly higher than that in the healthy control group(413. 6 3 ± 330. 69;t= -2. 497,P﹤0. 05). And miRNA-16 expression in SLE active group was 1 298. 79 ± 803. 79,significantly higher than that in SLE stable group(540. 95 ± 350. 15;t= -2. 445,P﹤0. 05). The level of miRNA-16 was related with AnuA (r=0. 669,P=0. 005),ESR(r=0. 608,P=0. 012)and SLEDAI(r=0. 530,P=0. 035). Conclusion The expression of miRNA-16 is high in SLE patients and it is related with SLE activity.

BACKGROUND: Percutaneous vertebroplasty for osteoporotic vertebral compression fractures has achieved very good results, but its long-term efficacy as well as impact on patients has been rarely reported so far.OBJECTIVE: To investigate the long-term effect of vertebroplasty with bone cement on osteoporotic vertebral compression fractures through a follow-up.METHODS: Thirty-four patients with osteoporotic vertebral compression fractures who had undergone percutaneous vertebroplasty were recruited. Visual analogue scale scoring was measured and compared as well as lesioned vertebral height and kyphosis angle shown on lateral X-ray examination prior to, 1 week and 6 years after percutaneous vertebroplasty.RESULTS AND CONCLUSION: The kyphosis angle was improved 1 week and 6 years after percutaneous vertebroplasty, and it changed insignificantly during the follow-up period. The vertebral height was also improved significantly after percutaneous vertebroplasty (P < 0.01); however, there was no obvious variation in the vertebral height at 1 week and 6 years after percutaneous vertebroplasty. The visual analogue scale exhibited an improvement after percutaneous vertebroplasty (P < 0.01); however, with time going by, the scoring on the visual analogue scale had an increased tend. All the parameters remained stable and had no large fluctuations. It is proved that the percutaneous vertebroplasty is effective and safe to treat osteoporotic vertebral compression fractures with an excellent long-term effect.

Objective To study plasmid-mediated transfer,plasmid replicon typing,and genetic environment of blaNDM-1 gene in Enterobacteraerogenes(E.aerogenes).Methods E.aerogenes HN-NDM0711 was used as the subject of this research,the transferable properties of plasmid were analyzed by conjugation testing,conjugant was performed stability testing,plasmid type was determined by PCR-based replicon typing (PBRT),downstream and upstream of blaNDM-1 were sequenced using chromosome walking method,genetic context was analyzed by BLASTN and BALSTP,as well as annotated using Vector NTI 11.5.1 software,sequence pipeline graph was made,the sequence was submitted to Genbank through software Banklt.Results The conjugation testing of E.aerogenes pHN-NDM0711 was positive,after positive conjugant was conducted 4-day passage,minimal inhibitory concentrations (MICs) of imipenem and meropenem to all the cloned strains didn't change,blaNDM-1 were all positive.The replicon type was IncA/C;blaNDM-1 gene was localized between ISAba14 and IS91,at upstream of the blaNDM-1,class 1 integron and Tn3 transposon were identified,class 1 integron contained a new mosaic structure of a drug-resistant resistance gene cassette.Conclusion E.aerogenes pHN-NDM071 1,bearing blaNDM-1 gene in IncA/C plasmid,derived from gene recombination under different antimicrobial selection pressure.Antimicrobial use in clinical,industrial and agricultural area should be strictly controlled,so as to reduce the emergence of such bacteria.

AIM:To explore an ideal method to induce the differen-tiation of human umbilical cord mesenchy-mal stem cells (hUCMSCs) into neuron-like cells and to provide some evidence for the transplantation of hUCMSCs for spi-nal cord injury .METHODS:The hUCMSCs were isolated from human umbilical cord digested with collagenase Ⅱ.The hUCMSCs was verified by flow cytometry analysis .The passage 5 cells were randomly divided into 4 groups.The differentiation of hUCMSCs was induced by bFGF in group A , bFGF and BDNF in group B, or BHA, bFGF and BDNF in group C, while the cells in group D served as a control group cultured with DMEM-F12 and 10%FBS.Two weeks later , the expression of nestin , neurofilament protein H ( NEFH) and glial fibrillary acidic protein ( GFAP) was detected by real-time PCR and immunocytochemistry .The morphological changes of cells were observed under an atomic force microscope . RESULTS:Mesenchymal stem cells were isolated and cultured from human umbilical cord by enzyme digestion .hUCMSCs expressed CD29, CD44 and CD105, but no CD34, CD45 or HLA-DR.After cultured with inducing medium for 2 weeks, the cells were successfully induced into neuron-like cells.The appearance of the cells had great change .The induced hUC-MSCs developed round cell bodies with multiple neurite-like extensions observed under an atomic force microscope .The re-sult of real-time PCR showed that nestin was positive in A , B and C groups , and NEFH was positive in A and B groups , but GFAP was negative in 4 groups.The difference of nestin and NEFH expression among the induced groups was signifi -cant (P<0.05).CONCLUSION:Mesenchymal stem cells were isolated and cultured from human umbilical cord by en-zyme digestion in vitro, and all the hUCMACs presented stable biological properties .Moreover, hUCMSCs were induced to differentiate into neuron-like cells in vitro via bFGF combined with BDNF .

Objective To evaluate the efficacy and safety of 0.05% desonide cream in the treatment of patients with eczema. Methods A randomized, double-blind, multicenter, vehicle-controlled study was conducted. The patients of the study and control groups applied 0.05% desonide cream and vehicle respectively, twice daily for 3 consecutive weeks. The efficacy was determined by measuring the total scores of erythema, erosion, infiltration, papule, exudation/crust, pruritus and the extent of lesions. Results At the end of the 3 weeks study, the total clinical effective rate was 80.8% in the study group,compared to 41.1% in the control group, with a significant difference between the two groups (P