Objective To study the clinical features ,the high risk features ,the treatments and the outcomes of chronic obstruc-tive pulmonary disease(COPD) patients combined with invasive pulmonary aspergillosis(IPA) .Methods The clinical dates of 48 COPD patients combined with IPA were retrospectively analyzed .Results The clinical manifestations of 48 COPD patients com-bined with IPA were dyspnea ,lung rales ,chest pain ,cough ,cough phlegm ,fever .Radiological findings were presented as pulmonary inflammatory exudation ,consolidation ,cavity of lungs .The treatments were effective in 16 cases ,ineffective in 32 cases .Conclusion Early diagnosis and treatment is important in COPD patients combined with IPA .

[ ABSTRACT] AIM:To study the influence of insulin resistance on fatty liver in the mice fed with high-fat diet (HFD).METHODS:Male 8-week-old C57BL/6J mice were randomly divided into HFD group (with 60% calories by high saturated fatty acid) and control group (with chow diet).The mice in both groups were fed for 12 weeks.The body weight, liver weight, serum triglyceride (TG) and total cholesterol (TC), and blood glucose and insulin levels were meas-ured.Hyperinsulinemic euglycemic clamp experiment was applied to reflect insulin sensitivity .The lipid deposition in the liver was analyzed by HE staining , Sudan IV staining and measurement of liver fat content .The phosphorylation levels of IRS1 and Akt, and the protein levels of SREBP-1 and FAS were determined by Western blot to reflect the activities of insu-lin signaling and lipid synthesis .RESULTS:Compared with control group , the body weight and liver weight were signifi-cantly increased in HFD group .TG and TC contents in serum and liver tissues were remarkably increased in HFD group . High-fat diet induced insulin resistance , as evidenced by increased serum insulin levels , reduced glucose infusion rate and decreases in IRS1 and Akt phosphorylation levels .In livers of HFD group, HE staining showed that the cytoplasm of hepa-tocytes was filled with vacuoles .Sudan IV staining also displayed that many different sizes of red lipid drops existed in the hepatocytes , and the protein levels of SREBP-1 and FAS were significantly increased .In primary normal hepatocytes with exogenous oleic acid intervention for 48 h, the phosphorylation levels of IRS 1 and Akt were reduced , and the protein ex-pression of SREBP-1 and FAS was significantly increased in a dose-dependent manner .CONCLUSION: Feeding with HFD leads to insulin resistance , resulting in activation of lipid synthesis and accumulation of lipid deposition in the liver , thus inducing fatty liver .

To establish a better method of primary culture for alveolar epithelial type II cells (AEC II) and to study its bionomics, alveolar epithelial type II cells were isolated by digestion with trypsin and collagenase, which were then purified by plated into culture flask coated with rat immunoglobulin G. The purified AEC II were identified by alkaline phosphatase staining, electron microscopy, immunocytochemical staining of pulmonary surfactant protein A (SPA). The SPA expression and transfection characteristics were compared with those of A549 cell line. The results showed that AEC II could be isolated by digestion with trysin and collagenase and purified by adhesive purification by using IgG, with a yield of about 2-3 x 10(7), and a purity of about 75%-84%. Cells could be quickly identified with AKP staining. AEC II were different from A549 cell line in terms of SPA expression and transfection characteristics. It is concluded that adhesive purification with IgG can improve the purity of AEC II, and AKP staining is simple in cell identification. AEC II can not be completely replaced by A549 cells in some studies because the differences between them, such as SPA expression.
Cell Culture Techniques/*methods ; Cell Separation/*methods ; Cells, Cultured ; Ecology ; Epithelial Cells/*cytology ; Immunoglobulin G/pharmacology ; Pulmonary Alveoli/*cytology ; Pulmonary Surfactant-Associated Protein A/biosynthesis ; Rats, Wistar

2,4,6-Trinitrotoluene (TNT) and its by-products dinitrotoluene (DNT) pose a significant threat to human health and other living organisms.However, the conventional analytical methods involved in bulky and expensive instruments are complicated and time-consuming, impeding quick and on-line determination.In this work, a facile yet effective strategy of utilizing UV-vis spectroscopy coupled with partial least squares (PLS) was proposed, through which TNT and two isomers of DNT (2,4-DNT and 2,6-DNT) in nature water could be rapidly and simultaneously determined without any pre-separation.Variable combination population analysis (VCPA) was utilized to select important feather variables and significantly improved the predictive performance of the PLS model.The calibration set contained 25 samples constructed by orthogonal array design (OAD).The predictive ability of the models was validated by an independent prediction set including 15 samples, achieving up to 0.99 of the determination coefficients (R2) for each of the analytes.The optimized models were successfully applied to determine the 3 ingredients in 8 environmental samples involving in tap, lake and two kinds of river water with the recovery values of great than 97%.Finally, the proposed method was further validated by high performance liquid chromatography method.UV-vis spectroscopy coupled with chemometrics may be used as simple and effective strategy with high potential in environmental monitoring.

Objective To investigate the effects of ulinastatin on pulmonary and hepatorenal function of elderly patients undergoing resection of esophageal cancer. Methods The clinical data of 40 elderly patients with esophageal cancer who had been admitted to Southwest Hospital from January 2005 to January 2008 were retrospectively analyzed. All the patients were randomly divided into control group (n = 20) and ulinastatin group (n = 20) according to random number table. Patients were administered with total parenteral nutrition, and patients in ulinaslatin group were additionally instilled with 4×10~5 U/d of ulinastatin. The levels of PaO_2, PaCO_2, PaO_2/FiO_2, alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (Tbil), blood urea nitrogen, creatinine and uric acid were detected preoperatively and at postoperative day 2 and day 6. All the data were analyzed by t-test and chi-square test. Results The levels of PaO_2 and PaO_2/FiO_2 were (87.3±4.2) mm Hg (1 mm Hg =0.133 kPa) and (416±20)mm Hg in ulinastatin group, which were significantly higher than (79.0±4.3)mm Hg and (376±20)mm Hg in control group (t =6.2, 6.2, P <0.05). The levels of ALT, AST and Tbil were (23±7)U/L, (38±8)U/L and (13.4±3.0) μmol/L in ulinastatin group, and (39±8) U/L, (50±9) U/L and (24.5±6.0) μmol/L in control group, with significant difference between the 2 groups (t = 7.0, 4.4, 7.6, P < 0.05). The levels of uric acid, blood urea nitrogen and creatinine were (279±84) μmol/L, (4.1±1.7) mmol/L and (66±12) μmol/L in ulinastatin group, and (386±67)μmol/L, ( 8.9±2.7) mmol/L and (95±38) μmol/L in control group, with significant difference between the 2 groups (t = 4.4, 6.4, 3.3, P < 0.05). The recurrence of complications in ulinastatin group was significantly lower than that in control group (χ~2 = 4.8, P <0.05). Conclusion Postoperative supplementation of ulinastatin and total parenteral nutrition is helpful in improving the pulmonary and hepatorenal function of elderly patients undergoing resection of esophageal cancer.

AIM:To clone the SPA gene promoter and construct its luciferase report vector of SPA gene and to study its transcriptional targeting activity.METHODS:① The SPA gene sequence was acquired from GenBank,of which the upstream was analyzed according to bioinformatics.The results showed that the upstream region of SPA gene sequence about 163bp has the function of promoter.② The SPA gene promoter fragment was generated by polymerase chain reaction and then subcloned into the multiple clone site(MCS)of luciferase report gene vector pGL3-basic to generate the recombined plasmid pGL3-SPA.This fragment was also subcloned into pGL3-control to generate recombined plasmid pGL3-SPA-enhancer by replacing its primary SV40 promoter.③ pGL3-SPA,pGL3-SPA-enhancer,pGL3-control,pGL3-basic were cotransfected with pRL-TK into A549 cells and H441 cells.The luciferase activities were measured by dual luciferase reportor(DLR)system.RESULTS:Sequencing and restricted digestive results showed that SPA gene promoter was successfully cloned and identified,and also correctly subcloned into plasmid pGL3-basic and pGL3-control to construct its luciferase report plasmid pGL3-SPA and pGL3-SPA-enhancer,respectively.The transcriptional activity was high in H441 cells.CONCLUSION:The luciferase report system of SPA gene promoter is successfully constructed with high transcriptional activity.

Objective To construct the eukaryotic expression vector harboring the fragment of Alia gene, and to investigate the effects of it on the signal of quorum sensing and virulence factors producted by Pseudomonas aeruginosa(Pa). Methods The plasmid pET-AiiA was cutted by Nhe Ⅰ and Xho Ⅰ , then the AiiA fragment was cloned into eukaryotic expression vector pEGFP-N2. After the plasmid was transfected into A549 cells, the protein was extracted and AiiA protein was found in it by Western blot. After the extrac- tion was admixed into the LB broth, from culture supernatant extracts of Pa, the N-acylhomoserine lactone (AHL) was detected by bioassay, and the expression of pyocyanin and elastase were assayed by RT-PCR and optical density. Results The fragment of AiiA gene was cutted and then cloned into pEGFP-N2. AiiA protein was found in the transfected cells. After admixed with the extract harboring AiiA protein, in Pa medium, the AHL was hydrolyzed, and the expression of pyocyanin and elastase were reduced. Conclusion The virulence factors synthesized by Pa were reduced by the AiiA protein expressed in eukaryotic cell.

AIM:To investigate the influence of calorie restriction ( CR) on contractility and mitochondrial bi-osynthesis in different types of rat skeletal muscles .METHODS:CR rat model was set up by feeding 60%normal food in-take of control rat every day for 8 weeks.Soleus (SOL) and extensor digitorum longus (EDL) were isolated under anesthe-sia.The twitch tension, titanic tension and fatigue resistance of SOL and EDL in response to electrical stimulation were measured to reflect the contractile function of the muscles .The copy number ratio of mitochondrial gene cytochrome C oxi-dase subunit I ( COX I) to nuclear gene β-actin was determined to evaluate the mitochondrial biosynthesis .ATP content was measured to mirror mitochondrial function .RESULTS:Compared with control group , CR for 8 weeks significantly in-creased twitch tension and titanic tension of both SOL and EDL , but only improved fatigue resistance in SOL .Markedly in-crease in ATP content in both skeletal muscles by CR intervention was observed , especially in SOL .Although CR activated AMP-activated protein kinase (AMPK) in both 2 muscles, up-regulation of mitochondrial biosynthesis and transcription of mitochondrial regulatory genes peroxisome proliferator-activated receptor γcoactivator 1α( PGC-1α) and nuclear respirato-ry factor ( NRF) was only observed in SOL .CONCLUSION:CR for 8 weeks enhanced the contractility of both rat SOL and EDL in response to electrical stimulation , especially in SOL composed of slow-twitch fibers.The mechanisms may be related to the activation of AMPK and the promotion of mitochondrial biosynthesis in SOL .

Objective To study the clinical isolation situation of Enterococcus bacteria and the drug resistance change to provide the basis for guiding clinical rational drug use and controlling infection.Methods The clinical specimens in our hospital from Janu-ary 2008 to June 2013 were performed the routine bacterial culture,identification and drug sensitivity test.The WHONET 5.6 soft-ware was used to analyze the distribution of the clinical Enterococcus bacteria l isolates in various samples and the departments and the drug resistance change during this period.Results Total 420 strains of Enterococcus bacteria were isolated during this period, including 288 strains of E.faecium(68.6%),128 strains of E.faecalis(30.5%),3 strains of E.gallinarum(0.7%)and 1 strain of E.casseliflavus(0.2%).The Enterococcus bacteria isolates were mainly distributed in urine(60.5%),wound secretion(20.0%) and blood(13.6%).The large changes occurred in the strains and drug-resistance of isolated Enterococcus bacteria during this peri-od.The proportion of E.faecium maintained about 65% every year.The proportion of E.faecalis was about 35%.The resistance rate of E.faecium to ampicillin,penicillin and nitrofurantoin was significantly higher than that of E.faecalis .Its resistance rates to penicillin,tetracycline,nitrofurantoin,high concentration gentamycin,high concentration streptomycin,ciprofloxacin and ampicillin were close to or over 70%.Total 3 strains of vancomycin-resistant E.faecium were isolated,including 2 strains in 2008 and 1 strain in 2009.Total 13 strains of vancomycin-intermediate E.faecalis were isolated,including 1 strain in 2008,5 strains in 2009 and 7 strains in 2010.Conclusion The drug resistance of Enterococcus bacteria is very serious and the drug resistance of different strains is different.The clinical anti-infection should rationally select the antibacterial drugs for increasing the curative effects according to the in vitro susceptibility of the isolated strains to the bacterial drugs.

Objective To investigate the therapeutic indications,dosage and prognosis of dermato-mucous membrane heamangioma(DMMH)in infants using low-dose superficial X-ray irradiation.Methods The patients were treated with a superficial X-ray machine(DERMOPAN typeⅡ)at voltages of29kV,43kV and50kV according to the depth and position of angioma,one or two times per week,1~2Gy per time,with the average total doses of5~10Gy and the maximal dose of20Gy.Results In a total of267patients,230cured,24markedly improved,10improved,and3failed.The cure rate was86.14%,and the total improve-ment rate was95.13%.Conclusion Treatment of DMMH with low-dose superfical X-ray is simple,conve-nient,safe,painless and cheap.The method has a high cure rate and is well accepted by the chlidren and their parents.