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.

Thirty-four patients with chronic lingual tonsillar hypertrophy were treated in our department from 1994 to 2000. There were 18 males and 16 females ranging in age from 21 to 60 years old. The clinical feature of lingual tonsillar hypertrophy usually appeared a series of pharyngeal symptoms, such as foreign-body sensation, sore throat, speech change, difficulty in swallowing, dysapnea and obstructive sleep apnea. The CO2 laser surgical lingual tonsillectomy was used for these patients. Result showed that the cure rate was 100% without any serious complications by following up 2 months to 3 years. The variety of presentations of lingual tonsillar hypertrophy and methods of surgical treament were discussed.

Breast Neoplasms ; diagnosis ; therapy ; Chemotherapy, Adjuvant ; Combined Modality Therapy ; Female ; Humans ; Neoadjuvant Therapy

Objective To improve the quality of antibiotics prescriptions in outpatient and emergency departments, to reduce the irrational use of antimicrobial drugs and to improve the quality of pharmacy service through system of rational drug administration.MethodsAnalyzing and evaluating the data of irrational administration of antimicrobial drugs between 2014 and 2015.The data was collected by the system of rational drug administration.ResultsFrom 2014 to 2015, the qualified rate of antibiotics prescriptions increased from 95.40% to 98.71%.Different types of irrational use of antimicrobial agents significantly reduced.ConclusionThe application of system of rational drug administration can improve the qualified rate of antibiotics prescriptions in outpatient and emergency departments and can promote pharmacy service.

Objective To study the effect of MK801 on behavioral changes and the possible mechanisms. Methods To observe the behavioral changes of levodopa induced dyskinesia (LID) rats during the period of chronic MK801 treatment, immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to measure the changes in expression of FosB, preproenkephalin (PPE) mRNA and prodynorphin (PDyn) mRNA in striatum, respectively. Double labling technique including immunohistochemistry of FosB and retrograde HRP transport tracing was used to observe the cell distribution of FosB. Results Pulsatile treatment with levodopa induced Abnormal involuntary movements (AIM) in PD rats, similar to LID in PD patients. FosB positive neurons and expressions of PPE mRNA and PDyn mRNA in striatum of 6-OHDA-lesioned hemisphere were increased in LID rats, and AIM scores of LID rats were reduced by MK801 treatment(41.9?15.6 vs 7.2?3.0), accompanied by the decrease in expressions of FosB and PDyn mRNA, but not PPE mRNA. Neurons immunoreactive for FosB were mainly located in striatonigral neurons which were labeled by cholera toxin-HRP (CT-HRP) injected in the substantia nigra pars reticulata (SNr). Conclusions MK801 could prevent the occurrence of dyskinesias induced by chronic levodopa treatment. The mechanism might be involved in the high expression of immediate early gene FosB and specific gene PDyn on the direct pathway. It suggests that LID might be related to the abnormal activity of direct pathway.

Objective To evaluate the effects of cardiac resynchronization therapy with optimized dynamic AV delay.Methods The study included 63 patients who underwent implantation of biventricular pacemakers for congestive heart failure.The dynamic AV delay was optimized on the day 1 week,6 months and 12 months after pacemaker implantation.The echocardiography and ECG,6MHW,QOL and NYHA grade were evaluated at the same time.Results The best optimized AV delay was between 90 ms and 140 ms(110.45?19.47 ms).Comparing the effectiveness 1 week after CRT with the day before CRT:the LVEF increased from 18.53%?4.83% to 22.07%?6.77% with the QRS interval decreased from 138.25?28.00 ms to 129.40?6.65 ms.Comparing the effectiveness of the day before and in 6-12 months after CRT optimized AV delay,the LVFT increased from 293.27?24.79 ms to 347.57?26.21 ms.There was no difference in QRS intervals.The LVEDD was upgraded from 71.43?7.12 mm to 57.06?6.54 mm while the QOL was upgraded from 74.14?6.24 to 55.15?11.02 and the NHYA upgraded from 3.12?0.55 to 1.82?0.43.Conclusion Optimized dynamic AV delay of the pacemaker parameters enhanced the short-term and long-term efficacy of CRT.

Objective To study the behavioral characteristics of a rat model of the levodopa-induced dyskinesia (LID) and the related factors, and to define clinically the relevant methods for assessing akinesia and dyskinesia in LID rats. Methods Unilaterally lesioned rat model of Parkinson′s disease using 6-hydroxydopamine were treated by levodopa and benserazide once daily for 3 weeks, on the 21st day the acute systemic administration of MK-801 was performed 15 min prior to levodopa treatment to observe the behavior (abnormal involuntary movement, rotation behavior and forelimb stepping) and to estimate the quality of AIM by using the rat AIM rating scale. Immunohistochemical technique was used to measure the number of TH-positive neurons in the substantia nigra (SN) and ventral tegmental area (VTA), which was then correlated to the AIM scores. Results Pulsatile treatment with a subthreshold dose of levodopa gradually induced abnormal involuntary movement (AIM), including stereotypy (limb dyskinesia, axial dystonia and masticatory dyskinesia) towards the side contralateral to the dopamine-denervated striatum and increased rotational behavior. The onset of AIM and motor pattern of each subtype was highly stereotypic across individual rats, and the proportion of each subtype was not consistent among individual rats. The number of TH-positive neurons in the VTA, but not in the SN, was significantly decreased in the LID rats compared with the non-LID rats. MK-801 prevented stereotypy but not rotational behavior. Contralateral forepaw performance was signi-ficantly improved after levodopa treatment, but gradually reduced with more and more severe AIM following repeated levodopa therapy. Conclusion Levodopa-induced rat AIM model of PD demonstrated similar properties with the levodopa-induced dyskinesia (LID) in PD patients, and provided an effective tool for LID study. AIM rating and forelimb stepping test are useful for evaluating the dyskinesia and akinesia of PD rats.

Objective To study the relationships between retinal microvascular disease and acute coronary event (ACE) among aged people. Methods A controlled study for senile people in communities was conducted. Xinglong Zhuang Coal Mine Community in Jining city, Shandong province was chosen to carry out the study, and the residents in that area aged≥60 years were asked to take questionnaire survey, physical and laboratory examinations. There were 139 cases met the diagnostic criteria of ACE being in the observation group, and 1 509 cases without ACE were assigned in the control group. The gender, age, smoking, alcohol intake, hypertension, diabetes mellitus, education, physical exercise, retinal microvascular disease, fasting blood-glucose, high density lipoprotein cholesterin (HDL-C), low density lipoprotein cholesterin (LDL-C), triacylglycerol (TG), systolic pressure, diastolic pressure, body mass index (BMI) were collected in the two groups to perform univariate analysis. Multivariate non-conditional logistic regression analysis was used for the factors with statistical significance to screen out the independent risk factors that could affect the occurrence of ACE. Results The univariate analysis showed:the risk factors that might cause the occurrence of ACE included age, gender, smoking, hypertension, diabetes mellitus, LDL-C, systolic pressure, diastolic pressure, BMI, and retinal microvascular disease (P<0.05 or P<0.01). In the ACE patients of observation group, the rates of presence of arteriovenous crossing sign [44.6%(62/139) vs. 27.8%(419/1 509)], hard exudates [9.4%(13/139) vs. 4.9%(74/1 509)] and cotton-wool patches [19.4%(27/139) vs. 7.3%(110/1 509)] in retinal microvascular disease were significantly higher than those in control group (P<0.05 or P<0.01). The logistic regression analysis showed:age [P=0.002, odds ratio (OR)=1.06, 95%confidence interval (95%CI)=1.04-1.09], smoking (P=0.032, OR=2.17, 95%CI=2.04-2.30), retinal microvascular disease (P = 0.010, OR = 2.33, 95%CI = 0.97 - 1.27), hypertension (P < 0.001, OR = 5.21, 95%CI=4.11-6.36), diabetes mellitus (P=0.021, OR=1.03, 95%CI=1.01-1.05) and LDL-C (P=0.020, OR=2.80, 95%CI = 2.65 - 2.99) were the independent risk factors for the occurrence of ACE. Conclusions Retinal microvascular disease is the independent risk factor for the occurrence of ACE. The retinal angiography can be a useful examination to forecast ACE.

Objective To study in patients with stable chronic obstructive pulmonary disease treatment, the use of Tiotropium Bromide Powder for Inhalation treatment combined with psychological intervention effect.Methods In 40 cases of patients with stable chronic obstructive pulmonary disease treatment using tiotropium bromide powder for inhalation, and classified as the control group, the other 40 patients in the control group based on the use of psychological intervention combined with treatment the treatment, and classified as the observation group, two groups of patients were in our hospital from January 2016 to January 2017.Results There was no significant difference in anxiety and depression of patients in the two groups before intervention, after the intervention group patients in the observation group improved significantly better;two groups of patients before intervention PaCO2, there was no significant difference in PaO2 index, after group intervention of patients in the observation group improved significantly better;compared two groups of patients with the treatment efficiency, the obvious observation group were higher, there were significant differences (P<0.05).Conclusion Tiotropium Bromide Powder for Inhalation combined psychological intervention treatment of patients with stable chronic pulmonary disease blocking was observed, which can improve the patient's condition, and improve the treatment effect, the patient's mental state adjustment also has a positive effect, it is worthy of reference.

The effects of solid-state fermentation (SSF) and submerged fermentation (SmF) and the addition of olive oil on the whole-cell lipase production by Rhizopus chinensis CCTCC M201021 were investigated.Compared with SSF, higher biomass, hydrolytic activity and synthetic activity were observed in SmF.By the addition of olive oil, the synthetic activity of whole-cell lipase in both fermentations was enhanced significantly, especially in SmF, while the biomass and the hydrolytic activity were also increased.Hence, olive oil serves as both carbon source and the inducer of lipases in fermentation.It was also found that the synthetic activity of whole-cell lipase was not accordant to the hydrolytic activity during both SmF and SSF, suggesting that isoenzymes with difference in lipase properties may be produced by Rhizopus chinensis.