Pigs are increasingly recognized as "natural" hosts of infection by human respiratory viruses because of their similarities to humans in terms of lung physiology, airway morphology, cell types, and distribution of cell receptors in the respiratory tract. We wished to explore the mechanisms of infection by respiratory viruses and screening of drug that could be used to treat respiratory-system diseases. Hence, we developed a model of well-differentiated porcine airway epithelial cells (PAECs) derived from pig-lung tissue and cultured them with serum-free medium under an air-liquid interface condition in vitro. We identified the PAEC model using scanning electron microscopy, electrophysiology, and immunohistology. To evaluate application of gene therapy of adeno-associated virus (AAV)6 on the PAEC model, we generated recombinant adeno-associated virus 6-green fluorescent protein (rAAV6-GFP) using the three-plasmid transfection method and infected PAECs from the apical surface with rAAV6-GFP. Results demonstrated that the PAEC model comprised a multilayer epithelial structure containing ciliated mucous secretory cells, with basal cells located directly beneath the multilayer. rAAV6-GFP could infect PAECs from the apical surface and efficiently transduce PAECs to mediate the long-term expression of the exogenous gene. Establishment of a model of well-differentiated PAECs in vitro could lay a solid foundation for the study of infection by respiratory pathogens, as well as the screening and gene therapy of agents used to treat diseases of the respiratory system.
Animals ; Cell Differentiation ; Dependovirus ; genetics ; Epithelial Cells ; cytology ; metabolism ; Green Fluorescent Proteins ; genetics ; HEK293 Cells ; Humans ; Lung ; cytology ; Membrane Potentials ; Mucins ; metabolism ; Swine ; Transduction, Genetic ; Tubulin ; metabolism

Objective To investigate the characteristics in sleep architecture of sub-healthy people with in-somnia,and to study the relationship between the sleep architecture and the degree of insomnia.Methods Sleep ar-chitecmre and Pittsburg sleep quality index(PSQI)value and PSQI scale were detected respectively.Results Sleep architecture of 46 subjects was abnormal-including shortened total-sleep-time(26.1%),excessive superficial-sleep stage(100%).inadequate deep-sleep stage(87.0%),insufficient rapid eye movement sleep(REM) (60.9%),longer sleep latency(65.2%)-more wakening times(47.8%)and longer wakeful time(43.5%).PSQI value of each insomniac exceeded 7,and the valtie of most objects was between 12 and 16(73.9%).The in-gredients of sleep architecture were not significantly correlated with the values of PSQI (P>0.05).Conclusion The sleep architecture of sub-healthy people with insomnia is mainly characterized with difficulty in falling asleep,light sleep and festless sleep,but the characteristics of sleep architecture is not inevitablly related with the degree of insomnia.

B ecause of the exist of com plex m atrix, the confirm ing indicators of qualitative results for toxic substances in biological sam ples by chrom atography-m ass spectrom etry are different from that in non-biological sam ples. E ven in biological sam ples, the confirm ing indicators are different in various ap-plication areas. T his paper review s the sim ilarities and differences of confirm ing indicators for the ana-lyte in biological sam ples by chrom atography-m ass spectrom etry in the field of forensic toxicological analysis and other application areas. T hese confirm ing indicators include retention tim e (R T ), relative re-tention tim e (R R T ), signal to noise (S/N ), characteristic ions, relative abundance of characteristic ions, parent ion-daughter ion pair and abundance ratio of ion pair, etc.

Objective:To study the effects on lymphocyte & T subset of SARS patients treated by integrative traditional Chinese and western medicine(ICWM). Methods:48 patients were randomly divided into two groups, the control group treated by western medicine (WM) alone and the test group treated by ICWM. The test group patients took the traditional Chinese medicine by 3 weeks based on their conditions. Lymphocyte and T subset were measured. Results:① At pretreatment, absolute value of lymphocyte of 38 patients in all patients were low. There were 19 patients with lowered CD3 +, and 28 patients with lowered CD4 +/CD8 + ratio in 38 patients with T subset measured. ② The value of lymphocyte in plasma in patients after ICWM treatment increased from 1 00?10 9?0 46?10 9 L -1 to 1 92?10 9?0 74?10 9 L -1 ( P

Objective:To evaluate the nutritional status before and after chemotherapy in patients with colon cancer by measuring energy metabolism and body composition.Methods:Fifty-one patients with colon cancer were included and were investigated using self-control method (before and after chemotherapy).Resting energy expenditure (REE) was determined by bed metabolic instrument,and body composition was measured by body composition analyzer.Results:For all patients,the REE on the 2nd day after chemotherapy was decreased significantly compared with that before chemotherapy (P ＜ 0.05).After chemotherapy,body fat percentage and body moisture percentage were increased significantly (P ＜ 0.05),and lean body mass percentage was decreased significantly (P ＜ 0.05).Conclusion:After chemotherapy,the REE and lean body mass percentage of the patients with colon cancer were decreased,and body fat percentage and body moisture percentage were increased.

Objective To investigate the risk of death between older and non-older critical patients in intensive care unit (ICU) in Shuyang People's Hospital.Methods A retrospective cohort study was conducted. The critical patients who aged 15 or above, and admitted to ICU of Shuyang People's Hospital from January 2014 to December 2016 were enrolled, and all the data was collected from theregistration and electronic medical records in the ICU. The prevalence and causes of death in ICU critical patients during the study period were observed. The patients were divided into elderly group (65 years and older) and non-elderly group (15-65 years), and logistic regression analysis was performed for the risk of death in the two groups.Results During the study period, 2707 critical patients in emergency were admitted to the ICU of Shuyang People's Hospital, and patients not satisfied the inclusion criteria were excluded. Finally, a total of 2466 patients were enrolled in the analysis with the male and female ratio of 1.6 : 1, an average age of (61.8±17.3) years, a median Glasgow coma scale (GCS) score of 6 (4, 8), and with a median ICU stay of 3 (1, 6) days. In 2466 critical patients, the most common cause of critical state was spontaneous intracerebral hemorrhage (25.5%) and traumatic brain injury (17.0%), with a fatality rate of 46.0% and 39.5% within first 7 days respectively. Compared with the non-elderly patients (n = 1415), the incidences of death of the elderly patients (n = 1051) due to traumatic brain injury, cerebral infarction, heart failure/cardiovascularcrisis, and respiratory critically ill were significantly increased (9.4% vs. 4.7%, 2.9% vs. 0.8%, 5.0% vs. 2.1%, 2.5% vs. 1.0%, respectively), while the incidence of death for pesticide/drug poisoning in the elderly group was significantly lower than that in the non-elderly group (0.2% vs. 1.2%, allP < 0.01). Stepwise logistic regression analysis showed that traumatic brain injury [hazard ratio (HR) = 1.878, 95% confidence interval (95%CI) = 1.233-2.864,P = 0.003), cerebral infarction (HR = 0.435, 95%CI = 0.229-0.826, P = 0.011), heart failure/cardiovascular crisis (HR = 0.399, 95%CI = 0.238-0.668,P = 0.000), and respiratory critically ill (HR = 0.239, 95%CI = 0.126-0.453,P= 0.000) in the older patients were significantly high risk factors of death as compared with those in non-older patients.Conclusions In the general ICU, the most common cause is spontaneous intracerebral hemorrhage and traumatic brain injury in critical patients with a high fatality rate. The risk of death in elderly patients with severe traumatic brain injury, cerebral infarction, heart failure/cardiovascular crisis, respiratory critically ill is higher than that of the non-elderly patients.

Objective To establish a method for rapid detection of JWH-018, JWH-250 and AM-2201 in blood by direct analysis in real-time coupled with tandem mass spectrometry. Methods These samples were extracted by acetonitrile-methanol (4:1), using DART 12Dip-it automatic sampling system. They were analyzed by positive ion and MRM mode. Results The detection limits of JWH-018, JWH-250 and AM-2201 were in good linearity in the range of 0.02-5.00μg/mL. The correlation coefficients were above 0.99 and the detection limits were 0.016μg/mL, 0.003μg/ mL and 0.017μg/mL, respectively. The intra-day and inter-day RSD were less than 15%. Conclusion The method has the advantages of high sensitivity and good accuracy. The sample processing is simple and can be analyzed in short time. This method is suitable for the analysis of JWH-018, JWH-250 and AM-2201 in practical cases.

Background Blindness and low vision represent significant public health issues in China.Late diagnosis is the major reason for the irreversible vision impairment.A feasible,cost-effective screening and referral program is very important for the eye health care,prevention and treatment of blindness in China.Objective This study was to evaluate the feasibility and effectiveness of a health examination center-based opportunistic eye disease screening program.Methods This was a cross-sectional study.Subjects undergoing a routine physical examination at the health examination center of the Fourth People's Hospital of Shenyang were invited to attend this program.Presenting visual acuity,intraocular pressure,and nonmydriatic fundus photography were obtained.Optic diso photographs were evaluated independently by two ophthalmologists.Blindness and moderate to severe vision impairment were defined based on the criteria of World Health Organization Visual Impairment Classification in 2009.Glaucoma,diabetic retinopathy (DR) and other suspected eye diseases were diagnosed according to the fundus photography and intraocular pressure.This study was approved by Ethic Committe of the Fourth People's Hospital of Shenyang,the informed consent of each subject was obtained.Results Totally,15 303 subjects were enrolled and 15 197 of them finished the exanimations,giving a response rate of 99.3％.The overall percentage of blindness and moderate to severe visual impairment was 0.08％ (12/15 197) and 2.34％ (355/15 197).Two hundred and twenty-eight (1.50％) subjects were defined as glaucoma suspects and 80 individuals (0.53％) were diagnosed as epimacular membrane.Other suspected eye diseases included DR (0.41％),branchial retinal vessel occlusion (0.24％),macular degeneration (0.09 ％),and macular hole (0.06％).More than 95 ％ of the eye disease suspects have never been previously diagnosed or treated.A total of 358 subjects (2.36％) were defined as ocular hypertension suspects.Conclusions This health examination center-based opportunistic eye disease screening shows a good efficiency and feasibility.It may become an optional program in the national eye health care project,as well as the work of prevention and treatment of blindness.

Objective To investigate the inhibitory effect of rapamycin,an mammalian target of rapamycin (mTOR) pathway inhibitor,on the proliferation,migration and fibrosis of human pterygium fibroblasts (PFBs). Methods Pterygium tissues were collected from patients with primary pterygium who underwent surgical excision in Shenyang Fourth People's Hospital from May to July 2015. The tissues were cultured in vitro and the PFBs were identified by anti.human vimentin immunofluorescence assay. The 3 to 5 generation cells were used for the experiments. The viability of cells treated with different concentrations of rapamycin was detected by methyl thiazolyl tetrazolium ( MTT) . The cells were divided into normal control group and rapamycin group, and the scratch wound healing test was used to evaluate migration of the PFBs. The expressions of MKI67,α.smooth muscle actin (α.SMA), fibronectin,caspase3, mammalian target of rapamycin ( mTOR ) and LC3B mRNA were detected by real.time quantitative PCR. Results The cultured cells showed morphology of long spindle and were vimentin immunopositive. The cell viability in rapamycin treated PFBs demonstrated a dose.dependent decrease. At 24 hours after culture,The cell viability in 30μmol/L rapamycin group was (76. 67±8. 84)％ of that in 0μmol/L rapamycin group ( P<0. 001 ) . The relative residual scratch width in 30μmol/L rapamycin group was ( 35. 40 ± 11. 62 )％ 48 hours after scratch,which was significantly greater than (2. 45±0. 76)％ in the normal control group (P<0. 05). Real.time quantitative PCR showed that the mRNA expressions of MKI67,α.SMA,fibronectin and mTOR in rapamycin group were significantly decreased when compared with those in normal control group (all at P<0. 05). The expression of LC3B mRNA in rapamycin group was significantly higher than that in normal control group (P<0. 05). The mRNA expression of caspase3 was not significantly different between the two groups (P=0. 861). Conclusions Rapamycin can effectively inhibit the proliferation, migration and fibrosis of PFBs without affecting the cell survival. Detailed mechanism remains to be further studied. Rapamycin may serve as an anti.fibrosis agent to prevent the progression and recurrence of pterygium in the future.

Objective To determine the spectrum of conjunctival flora and the antibiotic susceptibility profiles of patients scheduled for penetrating intraocular surgeries.Methods A prospective case control study was performed.A total of 192 patients (192 eyes) scheduled for penetrating intraocular surgeries at the Fourth People's Hospital of Shenyang from February to August 2015 were enrolled.Samples from the conjunctival sac were collected before instillation of any ophthalmic solutions for both aerobic and anaerobic culture.The positive rate and bacterial spectrum were observed.Bacterial isolates were tested for antibiotic susceptibility to 7 commonly used ophthalmic antibiotics using automated drug resistance analyzing system.The research was approved by the Ethics Committee of the Fourth People's Hospital of Shenyang.Results Totally 91 strains were collected from 81 conjunctival samples during aerobic culture,the positive rate was 42.19％.Staphylococcus epidermidis was the most common microorganism (64.84％),followed by Staphylococcus lentus (7.69％) and Staphylococcus aureus (3.30％).Coagulatase negtive Staphylococcus (CNS) accounted for 80.22％ of the positively cultured aerobes.For anaerobic culture,a total of 28 strains were isolated from 28 conjunctival samples,the positive rate was 14.58％ Propionibacterium acnes was the predominant species (71.43％),followed by Finegoldia magna (10.71％).Majority of the CNS were sensitive to gentamycin and vancomycin,with resistance rates lower than 10％,but their resistance rate to erythromycin and ceftazidime was 87.67％ and 63.01％,respectively.Resistance rate of these CNS to levofloxacin,ciprofloxacin,and moxifloxacin was 42.47％,39.73％ and 17.81％,respectively.Multidrug resistance to at least 3 antibiotic classes was present in 38.36％ of the CNS.Conclusions Bacteria in the conjunctiva sac of preoperative patients are resistant to various ophthalmic antibiotics.To follow-up the bacterial distribution and antibiotic resistance is great meaningful in the prophylactic and treatment in ocular surgery-related infections.