Objective To construct pEGFP-N1-TGF-β1 recombinant plasmid and transfect it into primary cultured neonatal piglet type Ⅱ alveolar epithelium cell (AEC-Ⅱ) by using lipofectamine 2000,in order to provide basis of methodology for producing recombinant plasmids for transplantation of transfected AEC-Ⅱ into ALI/ARDS animal model lungs.Methods PCR primers were designed to amplify the human TGF-β1 cDNA fragment from plasmid.XhoI and EcoRI were used for double digesting the empty plasmid pEGFP-N1 and cDNA fragment of human TGF-β1.Then the products of double enzyme digestion by using T4 DNA ligase were connected and transformed into DH5α and cultured over night for 16 hours.The structure of recombinant plasmid was identified by using PCR and base sequencing to verify the correctness of pEGFP-N1-TGF-β1 recombinant plasmid.It was then transfected into primarily cultured AEC-Ⅱ by lipofectamine2000 mediated transfection and cultured for another 48 hour.Plasmid DNA (pEGFP-N1-TGF-β1 recombinant plasmid) and lipofectamine 2000 were added into serum-free DMEM respectively,then DNA suspension and Lipofectamine 2000 suspension were blended together and added into cells.After 24-48 hours later,the expression level of enhanced green fluorescent protein (EGFP) was evaluated under fluorescence microscope.Results The structure of vector was verified as pEGFP-N1-TGF-β1 recombinant plasmid by using PCR and base sequencing.Green fluorescence found in some cells showed that the pEGFP-N1-TGF-β1 recombinant plasmids had been successfully transfected into primary cultured AEC-Ⅱ,however,the transfection efficiency still need tobe further improved such as repeating the transfection procedure once again or using adenovirus mediated transfection method to improve the efficiceny and to transplant the cells into animal lungs eventually.Conclusions pEGFP-N1-TGF-β1 recombinant plasmid was successfully constructed and,for the first time,transfected into primarily cultured AEC-Ⅱ of newborn piglets.This established method should be useful for investigation of therapeutic effect and outcomes of lung with experimental acute lung injury.

In the course of hospital regrouping it is imperative to integrate medical functions, resource elements and hospital cultures. Yet the integration of hospital cultures plays a pivotal role, for the culture of a hospital is the basis of its survival and development and cultural integration can lead to the fusion of the staffs feelings, values and concepts and to the coordinated development of the hospitals hardware and software, which will produce greater benefits. Ways of cultural integration include: ①doing a good job of intention surveys and ideological persuasion prior to hospital regrouping; ②adopting different forms of cultural fusion for different forms of regrouping: with unbalanced regrouping, the cultures of the weaker hospitals should not be totally negated and attention should be paid to equality and impartiality, whereas with balanced regrouping, efforts should be made to enhance communications rather than to differentiate the primary and the secondary; ③giving full play to the role of informal organizations within the hospital in the fusion of different cultures.

Objective To find out universal problems in restructuring and explore relevant methodologies so as to provide the government with decision-making basis. Methods The method of partial quantification from statistics and operational research and the qualitative method from sociology were employed. Results Hospitals A and B assumed the functions of a second-tier hospital and satisfied the basic medical needs of District Z. With Hospital A being superior to Hospital B and inpatient service superior to outpatient service, the obstruction to the restructuring lay in the staffs of the two hospitals. If the restructuring should turn out to be successful, there would be more space for the expansion of their share of the medical market. The focus of the restructuring should be put on the advancement of technologies and the improvement of quality and efficiency. In particular, the basic medical needs of the residents in the areas concerned must be met. Conclusion In restructuring two hospitals, the model of loose cooperation and steady combination is recommended in the first place while the government ought to play the role of providing guidance, coordination and policy support.

Objective To evaluate the effect of ?-lipoic acid(ALA)as a antioxidant on the histomorphologic changes of the kidney in type 2 diabetic Goto Kakisaki rats.Methods Four healthy Wistar rats(normal control group)and 10 GK rats were divided randomly into two groups:diabetic control group and lipoic acid-treatment diabetic group(?-lipoic acid 35 mg/kg intraperitoneal injection qd alt),who were both fed for 12 weeks.Indexes of oxidative stress of kidney were measured.Sections of renal tissue were evaluated using both a light and electron microscope to assess histomorphologic changes.Paraffin sections of the left kidneys were prepared and stained with H&E,PAS and Masson stains to compare the histomorphologic changes.The image analysis software of Motic Image Advanced 3.1 was used to measure the area of glomerulus and mesangial region,and the thickness of the glomerular basement membrane(GBM).Results Compared with normal control group,there were significant decreases in GSH,SOD,VitC and VitE levels in diabetic control group.The kidneys of the normal control group exhibited normal cortical morphology as determined by light microscope.Mild glomerulosclerosis,characterized by widening of the mesangial region,mesangial expansion and intercapillary cells proliferation,and GBM thickening under electron microscope were observed in the diabetic control group.In the ALA-treatment group,all the above pathological changes were alleviated.Conclusion In the kidney of type 2 diabetic rat,there exists oxidative stress which is associated with diabetic nephropathy.?-Lipoic acid can alleviate the abnormal histomorphologic changes of the kidney via antioxidative effects.

40 cases of parotidectomy were reviewed.Anterograde approach(group A)and retrograde approach(group B)for facial nerve dissection were used in 1 7 and 23 cases respectively.1 patients in group A and 8 in group B presented symptoms of temporary facial nerve paralysis(P <0.05).The anterograde method is safer and quicker.

Diagnostic Errors ; Encephalocele ; diagnosis ; Female ; Humans ; Meningocele ; diagnosis ; Middle Aged ; Mucocele ; diagnosis ; Sphenoid Sinus ; pathology

AIM:This paper described a HPLC method.Paeoniflorin and puerarin in Rg soft-exact(Radix Puerariae lobatae,Redix Paeoniae alba,etc.) were simultaneously determined.METHODS:A simplified HPLC procedure was developed.A 4.6 mm?250 mm Alltima C_(18) column with a mobile phase consisting of NaH_2PO_4(20 Mm,pH 2.4)-Acetonitrile:Tetrahydrofuran(900:120:5) was used.The flow rate was 1.00 mL/min.It was detected at 320 nm.RESULTS:Paeoniflorin and puerarin were well separated.The average recoveries of paeoniflorin and puerarin were 98.10% and 101.2% respectively.CONCLUSION:The result shows that this method is convenient,rapid and accurate.

OBJECTIVE To investigate the prevailing strains of pathogens of hospital acquired infections in ICU and their drug-resistances in order to provide treatment basis for critical and severe patients in fighting against infection.METHODS A retrospective investigation analysis was made for all the isolated bacteria as well as their drug-resistance in our hospital from Jan 2000 to Dec 2005.RESULTS Totally 869 bacterial strains were isolated which included 666 strains of G-bacteria(74.3%),and 230 strains of G+bacteria(25.7%).The G-bacteria included Pseudomonas aeruginosa,Klebsiella pneumoniase,Escherichia coli,Acinetobacter baumannii,etc,which were isolated mainly from respiratory tract.The G+ bacteria consisted of Staphylococcus and Enterococcus.Staphylococcus were mainly isolated from respiratory tract and Enterococcus from urinary tract.Bacteria were highly resistant to commonly used antimicrobials and demonstrated multi-drug resistance.The isolated rate of G+bacteria and drug resistance of G-bacteria to the commonly used antibiotics was increasing yearly.CONCLUSIONS The pathogens are mainly isolated from respiratory tract and the most are G-strains and multi-drug-resistant.The selection and use of antibiotics should be based on the results of drug-sensitivity tests.

Catheterization ; instrumentation ; methods ; Endoscopy ; Humans ; Paranasal Sinuses ; Sinusitis ; therapy

Objective To study the changes in tumor necrosis alpha(TNF ?), interleukin 6(IL 6), procalcitonin(PCT) and C reactive protein (CRP) levels and their values in the prediction and assisting diagnosis of infection complications in severe multiple trauma patients. Methods TNF ?, IL 6 and CRP were measured by enzyme linked immunosorbant assay(ELISA) and PCT was determined by immunoluminometric assay(ILMA) in serial samples of plasma from 34 patients with severe multiple trauma(ISS≥16) and compared with those of 11 normal control volunteers. Patients were divided into 2 groups(infection and non infection groups). The differences of the above parameters between groups were compared. In addition, all data were managed by receiver operating characteristic(ROC), and sensitivity, specificity, negative and positive predictive values for the diagnosis of infection complications were calculated. Results Plasma levels of TNF ?, IL 6, PCT and CRP in multiple trauma patients without infection complications were increased differently in the early posttrauma period. TNF ? and CRP levels in the early posttrauma period before infection were not significantly different from those in patients without infection complications, but they increased significantly following infection. However, during the whole observing period, plasma levels of PCT and IL 6 in the patients with infection complications were higher than those in patients without. The value of those parameters in assisting diagnosis of infection was evaluated. PCT was of the highest specificity and higher negative and positive predictive values. Conclusion All the parameters are of clinical value in infection diagnosis after severe multiple trauma. PCT is the best index for the prediction and assisting diagnosis of infection after multiple trauma.