Objective To survey the value of quantitative detection of urinary endotoxin for the diagnosis of gram-negative bacteria infection in urinary system,in order to provide evidences for early diagnosis and treatment.Methods The levels of urinary endotoxin were detected by kinetic turbidimetric technique,at the same time urine was cultured and WBC,CRP were detected.Results In 379 patients with gram-negative bacteria infection in urinary system,132 patients were upper urinary tract infection and 247 were lower urinary tract infection.The level of WBC,CRP and endotoxin were 11.25(7.98,13.99)× 109/L,81.36(76.64,83.50)mg/L and 49.57(17.28,78.37)pg/ml in upper urinary tract infection patients and in lower uri-nary tract infection patients were 10.24(7.94,12.24)×109/L,70.46(70.46,77.32)mg/L and 21.18(21.18,37.31)pg/ml. The group of upper urinary tract infection was higher than lower urinary tract infection on CRP and endotoxin,and have sig-nificant difference between the two groups of patients (P<0.05).Analysis of ROC,the diagnostic Cut-off value of 39.7pg/ml,the sensibility and specificity of endotoxin for the diagnosis of gram-negative bacteria infection on upper or lower urinary system were 84.1% and 90.3%.Conclusion The levels of urinary endotoxin detected by kinetic turbidimetric technique is simple and efficient,and have significant diagnosis value of gram-negative bacteria infection on upper or lower urinary sys-tem.

Objective To investigate the apoptosis response to mechanical stretch in human alveolar type Ⅱ epithelial cells A549.Methods First,A549 cells were given different stretch(0%,5%,15%,30%) at frequency of 0.5 Hz for 4 h.Then,A549 cells were stretched for different periods(0 h,2 h,4 h,8 h) at stretch 15%,0.5 Hz.Last,A549 cells were stretched for different frequency(0 Hz,0.2 Hz,0.5 Hz and 1 Hz) at stretch 15% for 4 h.The early apoptosis was detected by flow cytometry. Results A549 cells submitted to cyclic stretch caused the early apoptosis in a time-,stretch-and frequency-dependent manner.In details,at 0.5 Hz and 4 h stretch-dependence occurred,at 15% stretch and 0.5 Hz time-dependence occurred,and at 15% stretch and 4 h frequency-dependence occurred. Conclusion Mechanical stretch can induce the early apoptosis in human alveolar type Ⅱ epithelial cells A549,which may be one of the mechanisms of ventilator-induced lung injury.

Objective To investigate the value of color Doppler ultrasonography in early diagnosis and subsequent follow-up of the transient synovitis of the hip in children. Methods Color Doppler ultrasonography was performed on 80 children with transient synovitis of the single hip, and the characteristics of the blood signal changes in the synovial membrane of the hip joint were observed before treatment, 4 days and 8 days after treatment, while the normal hip joints were examined as the controls. Results Abnormalities in the injured side of the hip joints were displayed in all 80 children. Obviously increased blood signal of the injured side was detected with injured before treatment, but gradually decreased 4 days after treatment, and became similar to the healthy side after 8 days. Conclusion CDFI has an important role in monitoring changes of blood supply, and affords clinical application value in the therapeutic effect observation in transient synovitis of the hip in children.

In order to observe the therapeutic effect of azithromycin combined with IFN-γ on mouse toxoplasmosis and its impact on the cellular immune function of mouse, a total of 100 BALB/c mice were selected and divided into 5 groups, namely an infection control group (Group A) , azithromycin treatment group (Group B) , azithromycin combined with IFN-γ treatment group (Group C) , IFN-γ treatment group (Group D) and blank control group (Group E). The mice in Group A, B, C, D were infected by Toxoplasma tachyzoites through intraperitoneal injection and those in Group B, C, D were treated with relative drugs 24 h later for S days. The survival time of mice in each group and the levels of CD4 ~+ and CD8~+ T cells in blood were observed. The results showed that azithromycin combined with IFN-γ could improve the therapeutic effect of mouse toxoplasmosis and the cellular immune function of mice.

Objective To evaluate the effects of sevoflurane postconditioning on mitophagy during myocardial ischemia-reperfusion (I/R) in rats.Methods Forty-two pathogen-free adult male SpragueDawley rats, weighing 250-300 g, were randomly divided into 3 groups (n=14 each) using a random number table: sham operation group (group S) , I/R group and sevoflurane postconditioning group (group SP).Myocardial I/R was induced by 30 min ligation of the left anterior descending branch of the coronary artery followed by 2 h of reperfusion.In group SP, 2.4％ sevoflurane was inhaled for 15 min starting from the onset of reperfusion, while 33％ oxygen was inhaled in group I/R.The rats were sacrificed at the end of reperfusion, and the hearts were removed for measurement of myocardial infarct size (by 1％ 2, 3, 5 triphenyltetrazolium chloride) , expression of LC3 Ⅱ/LC3 Ⅰ , Beclin-1, p62 and Parkin (by Western blot) ,and mitochondrial menbrane potential (by using JC-1 probe) , and for examination of the uhrastructure of cardiomyocytes (with transmission electron microscope).Results Compared with group S, the myocardial infarct size was significantly increased, mitochondrial membrane potential was decreased, the expression of LC3 Ⅱ/LC3 Ⅰ , Beclin-1 and Parkin was up-regulated, and the expression of p62 was down-regulated in group I/R.Compared with group I/R, the myocardial infarct size was significantly decreased, the mitochondrial membrane potential was increased, and the expression of LC3 Ⅱ/LC3 Ⅰ , Beclin-1, p62 and Parkin was down-regulated in group SP.Conclusion Sevoflurane postconditioning can mitigate I/R injury in rats, and inhibition of excessive activation of mitophagy may be involved in the nechanism.

Objective To explore the effect of different method in preventing broncho-pulmonary dysplasia(BPD) in very low birth weight infants(VLBWI) during mechanical ventilation.Methods The gestational age of 71 infants with VLBWI was (30?2) weeks and the average weight was (1235.2?160.6)g.The infants were randomly divided into three groups,they received different therapeutic regimens during mechanical ventilation:32 cases were treated with permissive hypercapnia(PHC) ventilation in group Ⅰ;20 cases were treated by intratracheal instillation of pulmonary surfactant(PS) in group Ⅱ;19 cases were treated by inhalation of becotide(beclomethasone dipropionate,BDP) in group Ⅲ.The ventilator settings,blood gas analysis,clinical symptoms and therapeutic effect were statistically analyzed and compared.Results The peak inspiratory pressure(PIP) and ventilation rate(VR) of ventilator parameter were (18.3?1.6)cmH_2O and (35?5)bpm in group Ⅰ,there was significantly lower than that in group Ⅱ and group Ⅲ(P0.05).The incidence of BPD was 16%(5/32) in group Ⅰ,5%(1/20) in group Ⅱ and 5%(1/19) in group Ⅲ;The mortality rate of PHC group(group Ⅰ) was 6%(2/32) and 5%(1/20) in group Ⅱ.Conclusion The application of permissive hypercapnia,intracheal instillation of pumlonary surfactant and inhalation of becotide all can prevent effectively the occurrence of severe broncho-pulmonary dysplasia in VLBWI during mechanical ventilation.

Objective To investigate the mechanism of therapeutic action of dexamethasone on asthmatic mice by detecting the levels of IL-25 and IFN-γ in bronchoalveolar lavage fluid (BALF). Methods Balb/c mice with SPF grade were randomly divided into normal control group, asthma group and dexamethasone group. Asthma group and dexamethasone group were sensitized and challenged with ovalbumin ( OVA) . Dexamethasone group was intraperitoneally injected with dexamethasone one hour before challenging. The mice were executed 24 hours after the last challenge, and the HE stained pathological sections of the right lung were made. Pathological sections of lung were observed. BALF in the left lung was also collected. The total white blood cell count and absolute eosinophile ( EOS) count were observed, and the percentage of EOS was calculated. The levels of IL-25 and IFN-γwere measured with ELISA, and correlation analyses were made. Results The counts of total white blood cell and EOS, and the percentage of EOS were significantly higher in the asthma group than in the normal control group and dexamethasone group (P<0. 05). No differences were found between the normal control group and dexamethasone group. The IL-25 level was higher in the asthma group than in the normal control group and dexamethasone group (P<0. 05), and its level in the dexamethasone group was also higher than that in the normal control group. The IFN-γlevel was lower in the asthma group than in the normal control group and dexamethasone group (P<0. 05), while there was no significant difference between the normal control group and dexamethasone group. IL-25 was negatively correlated with IFN-γin each group. Conclusion Part of the mechanisms of dexamethasone acting on asthma are related to its inhibition on the pulmonary inflammation and promotion on the expression of IFN-γ, and possible inhibition of IL-25 expression.

Objective To observe the curative effect of autologous bone marrow stem cells implantation to bone inducing active material combined with core decompression in the treatment of early femoral head osteonecrosis (FHON).Methods From April,2010 to March,2012,in Department of Orthopaedics,the First Affiliated Hospital of Zhengzhou University,a total of 79 adult patients with 108 hips suffered from the early stage FHON were treated with autologous bone marrow stem cells implantation to bone inducing active material combined with core decompression through the core of the femoral canal,male of 65 cases,female of 14 cases,the mean age was 29.5 (20-50) years old.According to the etiology classification:the alcohol-induced FHON was in 54 patients with 66 hips,steroid-induced FHON in 14 patients with 20 hips,steroid and alcohol-induced ONFH was in 11patients with 22 hips.According to association research circulation osseous (ARCO)classifying,Ⅰ-A,Ⅰ-B,Ⅱ-A,Ⅱ-B phases were 6,16,8,and 78 hips,respectively.There were 43 hips in left side and 65 hips in the right side.Results All patients were followed up from 4 to 6 (4.8 ± 0.6) years.Compared with before operation,the scores of all patients were significantly increased (P ＜ 0.05).All patients with hip pain symptoms were relieved or disappeared.The healing tine of the patients in all age groups was statistically significant (P ＜ 0.05),and with the increase of age,the healing time was prolonged.The excellent and good rates of Ⅰ-A,Ⅰ-B,Ⅱ-Aand Ⅱ-B were 100％ (6 / 6),100％ (16/16),100％ (8/8),and 98.7％ (77/78).The X-ray showed that coarse channel osteogenic phenomenon is obvious,there is 1 case collapse of femoral head of stage Ⅱ-B,the rest were not collapse.Conclusion The treatment of early osteonecrosis of the femoral head with autologous bone marrow stem cells implantation to bone inducing active material combined with core decompressionis definitely effective,especially in patients with ARCO:Ⅰ-A,Ⅰ-B and Ⅱ-A phase,and the effect of ARCO:Ⅰ-A and Ⅱ-A is the best.

OBJECTIVETo investigate the effects of butyl alcohol extract of baitouweng decoction (BAEB) on the fungal cell surface hydrophobicity (CSH), filamentation and biofilm formation of Candida tropicalis.

METHODGradual dilution method was used to determine the MIC. XTT assay was applied to determine the SMIC80. Time-Kill assay was employed to draw the Time-Kill curve. The water-hydrocarbon two-phase assay was used to measure the cell surface hydrophobicity. Scanning electron microscopy (SEM) was applied to observe the morphological changes of the biofilm. Confocal laser scanning microscopy (CLSM) was applied to determine the thickness of the biofilm. The quantification real-time PCR (qRT-PCR) was used to detect expression changes of releated genes (UME6, ALST3 and NRG1). result: The MICs of BAEB against C. tropicalis strains are determined as 64-128 mg x L(-1). The SMIC80 s of BAEB against the biofilm of Candida tropicalis strains are determined as 256-512 mg x L(-1). Time-Kill curve results indicate that BAEB has a promise fungicidal effect at 256 and 512 mg x L(-1). SEM results shows that 512 mg x L(-1) BAEB can inhibit the formation of C. tropicalis biofilm on Silicone catheter, and the morphology of biofilm is also affected by BAEB. The thickness of C. tropicalis biofilm is reduced by BAEB according to CLSM results. Furthermore, qRT-PCR results indicate that expression of UME6 and ALST3 are significantly down-regulated by BAEB 256,512 mg x L(-1), and NRG1 is not affected by BAEB.

CONCLUSIONBAEB inhibits effectively the CSH, filamentation and biofilm formation of VVC strains of C. tropicalis.

Antifungal Agents ; chemistry ; pharmacology ; Biofilms ; drug effects ; Candida tropicalis ; drug effects ; genetics ; physiology ; Candidiasis ; microbiology ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Fungal Proteins ; genetics ; metabolism ; Gene Expression Regulation, Fungal ; drug effects ; Humans ; Virulence Factors ; genetics ; metabolism

BACKGROUND: At present, the commercial artificial small vascular grafts (diameter < 6 mm) are still unsatisfactory, due to poor blocompatibility and low long-term patency rate. Therefore, finding a vascular substitute with normal biological function and studying construction and function of tissue-engineered blood vessel have become hot topics recently.OBJECTIVE: To construct a novel tissue-engineered blood vessel by rabbit bone marrow mesenchymal stem cells (MSCs) and vascular acellular matrix, and to investigate the biocompatibility and patency rate of tissue-engineered blood vessels.DESIGN, TIME AND SETTING: An in vitro randomized controlled study at level of cytology and histopathology was performed at the Laboratory of Affiliated Drum Tower Hospital of Nanjing University Medical School from January 2006 to June 2008.MATERIALS: The decellularized vascular acellular matrix was obtained by a detergent-enzymatic procedure. MSCs from rabbits were isolated using density gradient centrifugation method and cultured in culture flasks coated with fibronectin. Subsequently, the expanded MSCs were seeded on the decellularized scaffolds, and then co-cultured in the self-made bioreactor to construct the tissue-engineered blood vessels.METHODS: Sixty rabbits were randomly divided into three groups. A1.0-cm abdominal aorta was sheared, and a tissue-engineered blood vessel was transplanted on the abdominal aorta using 8/0 polypropylene thread. Tissue-engineered blood vessel group: Tissue-engineered blood vessel was considered as the transplanted vessel; vascular acellular matrix group:Xenoma artery treated by vascular acellular matrix was considered as the transplanted vessel; xenoma artery group: Fresh xenoma artery was considered as the transplanted vessel.MAIN OUTCOME MEASURES: Immunocytochemical staining was used to identify the cultured MSCs. After 3 months of transplantation, the grafts were retrieved for digital subtraction angiography, pathological test and scanning electron microscope examination.RESULTS: Rabbits MSCs presented a whirlpool-like appearance at 8 days after culture. The immunocytochemistry results were consistent with the phenotype of MSCs. After high proliferation, MSCs were seeded onto the vascular acellular matrix for 12 days,and seed cells attached to well in the lumen of blood vessels. Three months after implantation, the patency rate was 90% of tissue-engineered blood vessel group and 80% of vascular acellular matrix group, which was superior to xenoma artery group (25%). At three months after transplantation, HE staining and scanning electron microscope demonstrated that internal, middle,and external membrane were clearly observed in the tissue-engineered blood vessel group, and the membrane morphology was similar to normal artery. The endothelial cells were covered completely. However, the endothelial cells were not covered completely in the vascular acellular matrix group, while mural thrombosis, mild proliferation of intima, and inflammatory cell infiltration were observed. The intima was thick and necrotic in the xenoma artery group, while lumens were stenotic and accompanied with a certain degree of thrombus organization.CONCLUSION: This study provides a new strategy to develop a tissue-engineered blood vessel with excellent biocompatibility and high patency rate constructed by rabbit MSCs and vascular acellular matrix.