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.

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.

ObjectiveTo investigate the independent risk factors of 30-day mortality of nontraumatic acute chest pain in emergency department so as to get non - traumatic acute chest pain risk score,MethodsThe clinical data of 532 patients with non - traumatic acute chest pain were reviewed.The independent risk factors of 30 - day mortality were identified after analysis of medical history,symptom and sign,laboratory findings by uuivariate analysis and logistic regression.Non- traumatic acute chest pain risk score was made as per the odds ratios of these risk factors. ResultsThe average age of the patients was (55.7 + 12.7 ) years,and 45 patients ( 8.4％ ) died after 30 days.In patients with non - traumatic acute chest pain,history of hypertension (OR:4.28; 95％ CI:1.59-11.55 ),prolonged chest pain (OR:1.1; 95％ CI:1.05-1.15),dyspnea (OR:6.61; 95％CI:2.40-18.10) and tachycardia (OR:1.02; 95％CI:1.00-1.04),high leucocyte count (OR:1.18; 95％CI:1.06-1.31) and D - Dimer ( OR:1.002; 95％ CI:1.001-1.002 ) predicted 30 - day mortality independently,whereas chest pain relieved by medicine (OR:0. 15; 95％ CI:0.04-0.65),high blood oxygen saturation (SaO2) (OR:0.89; 95％CI:0.83-0.98) and normal hematocrit (OR:0.92; 95％CI:0.86-0.99) were good markers to predict optimistic prognosis.Non - traumatic acute chest pain risk score was higher in 30 - day dead group than those in survival group significantly ( P ＜ 0.01 ),and mortality was significantly different between groups with various risk stratification (P ＜ 0. 01 ).Conclusions Clinical physician can predict 30 - day mortality and evaluate prognosis in patients with acute chest pain by using non - traumatic acute chest pain risk score quickly and effectively.

Objective:To analyze and evaluate the effect of OX40L as a potential adjuvant for H7N9 whole-virion inactivated vaccine (WIV).Methods:Fifty BALB/c mice were randomly divided into five groups and immunized intramuscularly with PBS (control group) and 1.5 μg WIV alone or in combination with 0.6, 1.8 or 3.0 μg Fc-fused OX40L (OX40L/Fc) adjuvant. Three weeks after immunization, IgG, IgG1 and IgG2 titers were measured by ELISA and hemagglutination inhibition (HI) assay. Moreover, the mice were challenged with 50×median lethal dose (LD 50) of homologous virus and the changes in mouse body weight and survival rate were recorded to evaluate the effects of OX40L. Flow cytometry was used to analyze the mechanism of OX40L as an adjuvant 7 d after immunization. Results:Compared with immunization with WIV alone, co-immunization of WIV with OX40L/Fc induced higher antigen-specific IgG in mice. The geometric mean titers (lgGMT) of antibodies induced by 0.6, 1.8 and 3.0 μg OX40L/Fc reached 3.79, 4.40 and 4.20, respectively. WIV combined with OX40L/Fc induced high levels of IgG1 and IgG2a without influencing Th1/Th2 balance. HI antibodies were also higher in WIV+ 1.8 μg OX40L/Fc and WIV+ 3.0 μg OX40L/Fc groups than in WIV group (6.25±0.50 and 5.70±0.97 vs 3.00±0.97, both P<0.05). WIV combined with 1.8 or 3.0 μg OX40L/Fc could protect 80% or 75% of mice against lethal challenge with H7N9 and result in less weight loss as compared with WIV alone. The most effective dose of OX40L/Fc was 1.8 μg. Flow cytometry showed that WIV (0.6, 1.8, 3.0 μg) in combination with OX40L/Fc enhanced the proliferation of T follicular helper cells (Tfh) through promoting the expression of CXCR5 and PD-1 as compared with WIV alone (all P<0.05). Conclusions:This study suggested that OX40L was beneficial to potent antibody responses induced by H7N9 WIV through promoting Tfh cell proliferation.

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 evaluate the applicability of activated charcoal in treatment of oral poisoning. Methods The feasibility of clinical manipulation and tastiness of 4 forms of activated charcoal were investigated, with different dosage forms in 35 nurses and 50 volunteers, respectively. The feasibility of the clinical manipulation was assessed by gastrelavage time and block numbers of stomach duet,and the tastiness by volunteers' taking orally. Results The gastrolavage time (151.8±17.8) s for powder, (96.9±24.80) s for suspension, (319.0±82.4) s for tablet and (314.3±93.3) s for suspension(P <0.001). Conclusion There are significant differences in the applicability of activated charcoal in different dosage forms. The suspension is the best form in feasibility. Capsule and tablet are better than powder and suspension in tastiness. Suspension and powder are the worst to accept.