Objective To observe characters of sputum of patients with tracheotomy by heat and moisture exchanger(HEM)undergoing hyperbaric oxygen therapy(HBOT)in multi-person chamber.Methods In randomly block design,239 patients were selected and received HBOT using HEM combined with L-shape tube(group HEM,n=128)and by L-shape tube(group L,n=111)respectively.The characters and volume of sputum,times of sucking sputum and cases of sealing/extubation of tubes were assessed over first and tenth sessions of HBOT.Results The times of sucking and volume of sputum over first session of HBOT in the group HEM were more than those in group L.The sputum was changing from ropiness over first session to tenuity over tenth sesion(P＜0.01).Cases with sealing/extubation of tubes in group HEM were more than those of group L(P＜0.01).Conclusions It is helpful to humidify and discharge deposited sputum of patients over routine hyperbaric oxygen by HEM.And the case8 of sealing/extubation of tubes are also reduced.

Objective To discuss the value of combining the physiological and operative severity score for enumeration of mortality and morbidity(POSSUM)and acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)in severe acute pancreatitis(sAP).Methods 60 cases of SAP were divided into surgery group(n=15)and non-surgical group(n=45),Non-surgical group was vahde with APACHE Ⅱ score,the surgery group with POSSUM score,decided to continue conservative treatment or surgery treatment,Observation of two ways with the complications and death.Results The rates of compHcafions and death were(0.37±0.08)and(0.27±0.09)in Single-operation group(n=15)higher than(0.76±0.14)and(0.61±0.15)in surgical group(n=11)(t1=3.125,t2=3.211,P＜0.01);APACHEⅡ score of mortality in operation group and non-surgical group were no significant difference(x2=2.28,x2=1.98,P＞0.05);APACHE Ⅱ score were(10.12±6.27)in survival group(n=46)were lower than (25.75±7.90)in death group(n=14)(t=2.525,P＜0.05.Conclusion The score of dynamic APACHE Ⅱ has better effect to judge the timing of surgery,and the score of POSSUM has high value to predict post-operative complications and deaths occurred in patients with severe acute pancreatitis.

Objective This study aims to investigate the clinical value of percutaneous vertebroplasty (PVP)combined with strontium-89 (89 Sr)on patients with spinal malignant tumor.Methods Seventy four patients who were diagnosed as spinal malig-nant tumor were divided into two groups randomly.37 patients of the control group were treated by PVP only,and 37 patients of joint group were treated by PVP and 89 Sr together.Then the clinical effect of the joint group and control group was compared after treatment.Results After treatment,the quality of life,ratio of pain relief,clinical effect and survival time in joint group were dis-tinctly better than control group (P <0.05).Conclusion PVP combined with 89 Sr can be an effective and safe treatment for pa-tients with spinal malignant tumor.

Objective:To investigate the clinical efficacy of fluorescence in situ hybridization(FISH)in prenatal diagnosis of chromosomal aneuploidy.Methods:FISH and karyotyping analysis were done in 120 samples of amniotic fluid from pregnant women.Results:FISH analysis results were consistent with those of karyotyping,the dectecting rate can reach 100%for chromosomal aneuploidies.There was no significant difference in the preference for FISH or karyotying for prenatal diagnosis when facing high matemal age,multi-indications and other factors(P>0.05).FISH was better when biochemical datafor down's syndrome were positive (P=0.029),however karyotyping was better when there was abnormal fetal ultrasound scan(P=0.000).Conclusions:FISH can detect the chromosomal aneuploidy quickly and accurately,especially when biochemical data for down's syndrome were positive.FISH could be the first choice for prenatal diagnosis in the third trimester of high-risk pregnancy.

ObjectiveTo purify and characterize the monoclonal antibody (McAb) against Chlamydia trachomatis pORF5 plasmid protein.Methods The hybridoma cells stably secreting specific McAb against pORF5 were cultured in a large scale,and protein G purification by affinity chromatography was used to purify 2H4 McAb.ELISA was used to determine the antibody titer,and identify McAb isotype.Immunofluorescence assay (IFA) and Western blot were performed to detect McAb specificity.Results The purity of 2H4 antibody was 93％,the titer reached 1:1024,and 2H4 McAb was identified to belong to IgG2a isotype,2H4 McAb reacted strongly with the GST-pORF5 fusion protein and endogenous pORF5 protein expressed by Chlamydia trachomatis serovar A,D,L2,Chlamydia muridarum ( MoPn ),Chlamydia psittaci 6BC,but not other chlamydial plasmid proteins and Chlamydia pneumoniae(Cpn) AR39 strain.Conclusion2H4 McAb against pORF5 protein was successfully purified with a high titer and specificity which lay a foundation for further study on pORF5 protein structure and function.

Objective To investigate the efficiency of antitumor immune responses induced by a controlled live dendritic cell(DC)vaccine Methods DC precursors were isolated from Fischer 344 rat bone marrow and cultured with granulocyte macrophage colony stimulating factor and interleukin 4 The rat ovarian tumor cell line NuTu 19 was genetically modified by retroviral mediated suicide gene(HSV 1 TK), and the positive clones were selected using G418 Live DC vaccine was then fused with DC and NuTu 19/TK cell by polyethylene glycol The characteristics of live DC vaccine were assayed with flow cytometry and confocal laser scanning microscopy The specific expression of HSV 1 TK gene in live DC vaccine was evaluated by RT PCR and western blot The sensitivity of live DC vaccine to ganciclovir (GCV) was evaluated by methylthiazoletetrazolium assay In vivo, rats vaccinated twice with live DC vaccine were compared to those vaccinated with killed DC vaccine, unfused DC and NuTu 19/TK cell or phosphate buffered saline Seven days following the last immunization, the rats were sacrificed to test the specific cytotoxic T lymphocyte (CTL) activity by lactate dehydrogenase release assay, or challenged with NuTu 19 and tumor incidence was observed Results The fusion efficiency was approximately (23?14) Live DC vaccine displayed an up regulated expression of major histocompatibility complex (MHC) IIOX6 (87 6?3 4)%, costimulatory molecule B 1 2 (71 1?9 3)%, integrin OX 62 (68 0?7 4)%, and adhesion ICAM 1 (77 1?2 0)%, and specifically expressed HSV 1 TK gene. Our data showed that spleen T lymphocytes from rats vaccinated with live vaccine displayed enhanced CTL aetivity (61 8?8 3)% contrast to that of rats vaccinated with killed vaccines (26 0?3 8)% ( P

Objective:To analyze the changes in biological characteristics including infectivity, growth and pathogenicity of Chlamydia muridarum ( Cm) after serial passage in vitro in special conditions in order to provide reference for screening attenuated live vaccines and virulence-related genes. Methods:Wild-type Cm strain (G0) was cultured for several passages using conventional cell culture method under alternate unassisted and assisted culture conditions. Then, the 28th generation (G28) of Cm was selected and compared with the parental G0 strain in terms of centrifugation dependence, attaching ability, intracellular growth curve, plaque size and fallopian tube lesions after genital tract infection in a mouse model. Results:Compared with the parental G0 strain, the G28 strain showed significantly decreased dependence on centrifugation during cell infection ( P<0.05) and increased attachment capacity to cells ( P<0.05). No significant differences were observed in the growth curves 32 h after cell infection or in the plaque sizes between the parental G0 and G28 strains. In the in vivo virulence test, fallopian tube lesions were observed in 87.5% of G0-infected mice and 37.5% of G28-infected mice ( P<0.05). Conclusions:Compared with the parental G0 strain, the G28 strain showed significantly enhanced in vitro infection ability, but decreased in vivo pathogenicity, which brought hope for further identification of virulence genes, isolation of attenuated strains with single genotype and development of live attenuated Chlamydia vaccines.

OBJECTIVETo investigate the implications of a prenatal diagnosis of single umbilical artery (SUA) for chromosomal abnormalities and neonatal outcomes.

METHODSFrom January, 2008 to June, 2012, color Doppler ultrasound identified 44 fetuses with SUA. Prenatal diagnoses with amniocentesis or umbilical blood sampling were subsequently ordered for routine chromosome karyotyping and the newborns were followed up for assessing the neonatal outcomes.

RESULTSOf all the 44 fetuses, 24 had uncomplicated SUA, and 20 had other concurrent abnormalities (including 8 with abnormal ultrasound soft indexes and 12 with chromosomal abnormalities). The two groups of fetuses showed significant differences in gestational weeks at delivery and incidence of chromosomal abnormalities but not in neonatal weight, placenta weight or APGAR score.

CONCLUSIONSFetuses with a prenatal diagnosis of SUA and other development abnormities need to undergo prenatal chromosomal examination. For fetuses with uncomplicated SUA, careful ultrasound examination is necessary to avoid missed diagnosis of potential congenital abnormalities.

Adult ; Chromosome Disorders ; diagnostic imaging ; genetics ; Female ; Fetus ; abnormalities ; Humans ; Karyotyping ; Pregnancy ; Pregnancy Trimester, Second ; Pregnancy Trimester, Third ; Single Umbilical Artery ; diagnostic imaging ; Ultrasonography, Prenatal ; Young Adult

Objective:To evaluate the role of histone demethylase (JMJD3) in drug-induced acute kidney injury (AKI) in mice.Methods:Twenty-four male C57BL/6 mice, aged 8-10 weeks, weighing 20-30 g, were divided into 4 groups ( n =6 each) using a random number table method: control group (C group), AKI group, a specific JMJD3 inhibitor GSKJ4+ control group (GSKJ4 group), and GSKJ4-AKI group.Folic acid 250 mg/kg was injected intraperitoneally to develop AKI model.GSKJ4 20 mg/kg was intraperitoneally injected at 1 h before developing AKI model in GSKJ4-AKI group and at the corresponding time point in GSKJ4 group.Blood samples were collected at 72 h after development of AKI model for determination of serum BUN and Cr concentrations.The animals were then sacrificed and renal tissues were collected for microscopic examination of histopathological morphology (using HE and PAS staining) and for determination of cell apoptosis (by TUNEL) and expression of JMJD3, Bax and cleaved caspase-3 (by Western blot), the number of JMJD3, myeloperoxidase (MPO), F4/80 and CD3 positive cells, expression of cleaved caspase-3 and Bax, and expression of IL-1β, IL-6, TNF-α and monocyte chemotactic protein 1 (MCP-1) mRNA (by reverse transcription polymerase chain reaction). The damage to the renal tubules was scored. Results:Compared with C group, the serum BUN and Cr concentrations and renal tubular damage score were significantly increased, the number of JMJD3, myeloperoxidase (MPO), F4/80 and CD3 + positive cells was increased, the number of apoptotic cells was increased, and the expression of Bax, cleaved caspase-3, JMJD3 and IL-1β, TNF-α, IL-6 and MCP-1 mRNA was up-regulated in AKI group ( P<0.05), and no significant change was found in the parameters mentioned above in GSKJ4 group ( P>0.05). Compared with AKI group, the serum BUN and Cr concentrations and renal tubular damage score were significantly decreased, the number of JMJD3, myeloperoxidase (MPO), F4/80 and CD3 positive cells was decreased, the number of apoptotic cells was decreased, and the expression of Bax, cleaved caspase-3, JMJD3, and IL-1β, TNF-α, IL-6 and MCP-1 mRNA was down-regulated in GSKJ4-AKI group ( P<0.05). Conclusions:The mechanism of drug-associated AKI may be related to up-regulation of JMJD3 expression and thus induces cell apoptosis and inflammatory responses in mice.

Objective:To evaluate the role of CXC chemokine receptor 6 (CXCR6)-mediated activation of natural killer T (NKT) cells in renal fibrosis following acute kidney injury (AKI) in mice.Methods:Eighteen male wild-type C57BL/6 mice and 18 CXCR6 knockout C57BL/6 mice, aged 8-10 weeks, weighing 20-30 g, were divided into 6 groups ( n=6 each) using a random number table method: wild-type mouse control group (group WT-CON), CXCR6 knockout mouse control group (group CXCR6 -/--CON), wild-type mouse with AKI group (group WT-AKI), CXCR6 knockout mouse with AKI group (group CXCR6 -/--AKI), wild-type mouse with AKI + NKT cell adoptive transfer group (group WT-AKI-NKT) and CXCR6 knockout mouse with AKI + NKT cell adoptive transfer group (group CXCR6 -/--AKI-NKT). Folic acid 250 mg/kg was intraperitoneally injected to establish the model of renal fibrosis in mice with AKI.NKT cellsuspension 250 μl(1×10 6 cells) was injected through the tail vein on the 4th and 9th days after folic acid injection in group WT-AKI-NKT and group CXCR6 -/--AKI-NKT, respectively.Blood samples were taken from orbital at day 14 after folic acid injection for determination of the concentrations of serum blood urea nitrogen (BUN) and creatinine (Cr). The animals were sacrificed, and renal tissues were obtained for observation of the area of renal fibrosis (by Sirius red staining) and renal injury (using H&E staining) which was scored and for determination of the proportion of CD1d Tetramer+ cells (by flow cytometry), the number of CD206 and α-smooth muscle actin (α-SMA) double positive (CD206 + -α-SMA + ) cells (by immunofluorescence) and expression of interleukin (IL)-4 and IL-13 mRNA (by real-time polymerase chain reaction). Results:Compared with group WT-CON, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased, and the expression of IL-4 and IL-13 mRNA was up-regulated in group WT-AKI and WT-AKI-NKT ( P<0.05). Compared with group WT-AKI, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased, and the expression of IL-4 and IL-13 mRNA was up-regulated in group WT-AKI-NKT ( P<0.05), and the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly decreased, and the expression of IL-4 and IL-13 mRNA was down-regulated in group CXCR6 -/--AKI ( P<0.05). Compared with group CXCR6 -/--CON, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased in group CXCR6 -/--AKI and group CXCR6 -/--AKI-NKT ( P<0.05). Compared with group CXCR6 -/--AKI, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased, and the expression of IL-4 and IL-13 mRNA was up-regulated in group CXCR6 -/--AKI-NKT ( P<0.05). Conclusion:CXCR6-mediated activation of NKT cells is involved in renal fibrosis following AKI in mice, and the mechanism may be related to promoting Th2 cytokine-mediated M2 macrophage-myofibroblast transformation.