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.

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 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.

In order to confirm the alteration and significance of cigarette smoke exposure on SP-A in rats, 20 Wistar rats were assigned randomly to two groups: an N group (n=10), and an S group (n=10). The ultra-structural change was observed by electron microscopy. The number of cells positive for SPA was by immunohistochemically measured. The mRNA expression in the lung tissues was determined by reverse transcription polymerase chain reaction (RT-PCR). The number of cells positive for SPA of the S group (0.52 +/- 0.05) was lower than that of the N group (0.72+/-0.06) (P<0.05). The levels of mRNA of SPA in the lung tissues of the S group (0.3522+/-0.0512) was significantly lower than that of the N group (0.4432+/-0.05628) (P<0.05). It is concluded that cigarette smoke alone decreased the level of SP-A and that might have an important effect on surfactant metabolism and the host defense functions of surfactant in the peripheral airways, which might play a crucial role in the development of chronic obstructive lung disease.
Gene Expression Regulation ; Immunohistochemistry/methods ; Lung/metabolism ; Microscopy, Electron ; Pulmonary Surfactant-Associated Protein A/*biosynthesis ; RNA, Messenger/metabolism ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Smoking/*adverse effects ; Tobacco Smoke Pollution/*adverse effects

Aim This study was aimed to explore the influence and mechanism of the long-term exercise on skeletal muscle contraction and mitochondrial biosyn-thesis in different muscle fibers.Methods Soleus (SOL)and extensor digitorum longus (EDL)were i-solated from SD male rats with platform running train-ing for eight weeks.The changes of contractility under different electrical stimulation were observed, mito-chondrial biosynthesis,including ATP content,mito-chondrial DNA,the gene expression of PGC-1αand NRF were also detected.Results Long-term endur-ance exercise can improve twitch tension and titanic tension of SOL and EDL ,but only enhanced the fa-tigue resistance in SOL.ATP contents were significant-ly increased in the two types of muscles,but mtDNA content,PGC-1αexpression and NRF translation were only obviously enhanced in SOL,in accompanied with an increase in p-AMPK/AMPK protein ratio.Conclu-sion Long-term endurance exercise increased skeletal muscle contractility and improved the anti-fatigue abili-ty in SOL,which may be associated with increase in mitochondrial biosynthesis via activated AMPK-PGC-1αaxis.

Objective To investigate the effects of calorie restriction (CR) for 4 weeks on twitch tension,titanic tension,and fatigue contraction induced by electrical stimulation in different kind skeletal muscles from rats and explore the possible mechanisms.Methods The rat model of CR was established by a limitation of 40％ calorie intake of control rats for 4 weeks,and then oral glucose tolerance test (OGTT) was performed.The soleus (SOL) and extensor digitorum longus (EDL) were isolated under anesthetization to detect twitch tension,titanic tension,and fatigue contraction induced by electrical stimulation.Adenosine triphosphate (ATP) content was measured by fluorescent enzymatic methods to reflect mitochondrial function.The ratio of mitochondrial gene COX Ⅰ and nuclear gene β-actin copy number was analyzed to evaluate mitochondrial biogenesis.Furthermore,the transcriptions of peroxisome proliferatoractivated receptor γ coactivator-1 (PGC-1α) and nuclear respiratory factor 1 (NRF1) genes,expressions of phosphorylated adenosine 5'-monophosphate-activated protein kinase (AMPK) and nitric oxide synthase (NOS) proteins,and nitric oxide (NO) content were determined in skeletal muscle.Results The blood glucose level at 30 min and area under the curve of blood glucose levels at various time points during OGTT were significantly decreased in the CR group compared to the control group.The twitch tension [(2.5 ± 0.15)N/cm2 vs (1.24±0.12)N/cm2,(2.66 ±0.21)N/cm2 vs (1.69 ±0.17)N/cm2,P ＜ 0.05],titanic tension [(10.43 ± 0.36) N/cm2 vs (8.06 ± 0.19) N/cm2,(11.35 ± 1.02) N/cm2 vs (8.12 ± 0.23) N/cm2,P ＜ 0.05],and fatigue contraction force in SOL and EDL from CR rats were significantly increased in association with increases of ATP content [(34.82 ±4.31)) mnol/mg protein vs (15.32 ± 1.94) nmol/mg protein,(30.82 ± 2.15) nmol/mg protein vs (12.32 ± 0.97) nmol/mg protein,P ＜ 0.05] and mitochondrial biogenesis (2.75 ± 0.20 vs 1.52 ± 0.06,1.32 ± 0.10 vs 0.84 ± 0.11,P ＜ 0.05) compared to control rats.CR for 4 weeks upregulated the transcriptions of PGC-1α and NRF genes as well as the phosphorylation of AMPK protein in SOL but not in EDL.Furthermore,CR also enhanced NOS expression and NO content in both skeletal muscles.Conclusions CR for 4 weeks can strengthen the contractile function of SOL and DL from rats,and the underlying mechanisms might be related to the upregulation of PGC-1α transcription and AMPK activation,resulting in the enhances of mitochondrial biogenesis and mitochondrial function in skeletal muscles.