Well-differentiated primary human airway epithelial cell culture (HAE), an organotypic human airway culture system, has been increasingly used as a linkage between researches conducted on animal models and humans.In this review, we focus on progress in technological development and its application in virology.The following aspects are covered in this review: (1)Structure and function of the normal airway, (2)Role of respiratory epithelial cells in pulmonary innate immunity, (3)Development of primary human airway epithelial cell cultures, (4)Application of primary human airway epithelial cell cultures in virology.

Animals ; Autophagy ; Coronaviridae ; genetics ; pathogenicity ; physiology ; Coronaviridae Infections ; physiopathology ; virology ; Humans ; Virus Replication

Objective To explore the effects of different levels of selenium nutrition on thyroid function,liver deiodinase type Ⅰ(DⅠ) and brain deiodinase type Ⅱ(DⅡ) activities in experimental autoimmune thyroiditis (EAT) rats. Methods A total of 32 female Lewis rats were randomly divided into four groups that included control group,model group,EAT with selenium-supplementation group and EAT with selenium-deficiency group. After two weeks of the basal diet administration,the rats were fed on forage containing different levels of selenium for nine weeks and the total intake of selenium were 4,4,40 and 0.4 ?g/d,respectively. The rats of model group,EAT with selenium-supplementation group and EAT with selenium-deficiency group were induced to establish the model of experimental autoimmune thyroiditis from the third week to eighth week. The pathological change of thyroid was examined,and the levels of selenium in the blood,serum thyroid autoantibody and thyroid hormone,DⅠ and DⅡ activities were measured simultaneously. Results Compared with control group,thyroid autoantibody and thyroid hormone levels significantly increased in model group,EAT with selenium-supplementation group and EAT with selenium-deficiency group (P

Objective To investigate the effects of rhBMP2 on the differentiation of human dental pulp stem cells and expression of Delta protein in vitro. provide a theoretical basis for research on human dental pulp stem cells. Methods Monocell suspension was separated from the human adult dental pulp with collagenase Ⅰ and dispase digestion.Clonogenic cells were observed under light microscope and the expressions of surface markers were determined with immunofluorescence. divided into experimental group (containing 50 ?g?L-1rhBMP2 in the culture medium) and negative control group (containing culture medium only).alkaline phosphatase and the expression of Delta proterin of the cells at at passage 5 were detected.Results The human dental pulp stem cells showed colony growth,the nestin and vimentin staining were positive by immunohistochemical staining.STRO-1 was positive in cells by immunofluorescence.Compared with negative control group, the activities of alkaline phosphatase increased after treatment with rhBMP2 for 7,14 and 21d(P

Objective To investigate the effect of glycosylphosphatidylinositol specific phospholipase D (GPI-PLD) on artherosclerosis.Methods The GPI-PLD activities and mononuclear cells GPI-PLD gene mRNA expression was detected in 102 patients with coronary artery disease and 121 healthy adult.The GPI-PLD highly expressing cell model was constructed and induced by ox-LDL,the HUVEC and HUVEC transfected with pcDNA3.1(+) as blank and control group,respectively.Before and after the induction,the change of cellular function and biological features was detected.Results The peripheral blood GPI-PLD activity of patients with coronary heart disease and normal control were 31.80±4.21 and 44.32±4.50,and the IA ratio of GPI-PLD mRNA expression of mononuclear cells were 0.92±0.16 and 1.53±0.14,respectively.The activity of GPI-PLD and mRNA expression in patients with coronary artery disease were decreased up to 28.2％ (t=21.31,P＜0.01) and 39.9％ (t=30.36,P＜0.01) as compared with healthy control.The adhesion cells,ET-1,reactive oxygen,malondialdehyde (MDA) and the apoptosis rate of GPI-PLD overexpressing HUVEC were lower as compared with blank [29.59=1.40,3.51 ± 0.45,(50.63 ±4.22) ng/L,0.043±0.011,(3.32±0.44) nmol/L vs.41.39±2.15,10.57±1.12,(59.35±4.45)ng/L,0.052±0.011,(5.01±0.69) nmol/L],and as compared with control [42.68±2.45,9.92±1.03,(61.11±4.12) ng/L,0.051±0.007,(4.89±0.71) nmol/l] after inducing by ox-LDL; while the level of nitrogen noxidium (NO) was higher than blank [(29.88± 1.37)μmol/L vs.(21.76±1.02)μmol/L] and control(23.43±0.85)μmol/L groups.Conclusions The expression and activity of GPI-PLD in patients with coronary artery disease are lower than health people.Stable high expression of GPI-PLD is beneficial to vascular endothelial cell injury repairment and prevent the occurrence and development of atherosclerosis.

The safety of simultaneous bilateral total knee arthroplasty (TKA) remains controversial. The objective of the current study was to investigate perioperative morbidity and mortality rates within 30 days of simultaneous bilateral TKA. A detailed analysis of medical, surgical and anaesthesia records of 183 consecutive patients who underwent total knee arthroplasty between 2002 and 2006 was performed. The mean age of the patients was 67.6 years old. More than 80% had one or more co-morbidities, but none of them had ASA score greater than class 2. The mean hospital stay was 10 days, and the mean surgical time 156 minutes. Less than half of the patients (42.6%) required blood transfusion. The rate of perimorbidity was 15.3 % and there was no mortality in this series. We believe that simultaneous bilateral total knee arthroplasty is a safe and cost effective option for our patients, provided that patients are selected and informed appropriately.

Objective To investigate the expression and clinical significance of zinc finger protein 217 (ZNF217)in human pancreatic cancer.Methods 43 cases with pancreatic cancer undergoing surgery in the PLA 117 Hospital and People's Hospital of Ruian City from Apr .2011 to May.2014 were enrolled in the study . The pancreatic cancer and the corresponding tumor-adjacent tissues were collected .Real-time quantitative PCR (qRT-PCR)was applied to detect ZNF217 mRNA expression in pancreatic cancer (n=43)and the corresponding tumor-adjacent normal tissues.The protein expression of ZNF217 was measured by immunohistochemistry(IHC). The relationship between the expression of ZNF 217 and clinical features was analyzed by pearson chi-square test . Results The expression level of ZNF217 mRNA and protein was significantly higher in pancreatic cancer tissues than in adjacent normal tissues(P<0.05).The high expression of ZNF217 protein was positively correlated with perineural invasion, tumor size, lymphatic metastasis and advanced TNM stage (P<0.05).Conclusions The expression of ZNF217 is significantly higher in pancreatic cancer tissues than in tumor-adjacent normal tissues , and the upregulation of ZNF 217 is associated with clinicopathological features of tumor malignance .ZNF217 may become a new marker and effective therapeutic target in early diagnosis of pancreatic cancer .

Objective To study the effects of glycogen synthase kinase-3?(GSK-3?)and free radical on neuron apoptosis of hypoxic-ischemic brain damage(HIBD)in newborn rats.Methods Eighty 7-day-old neonatal rats were randomly divided into 2 groups:normal group and hypoxic-ischemic(HI)group.Rats in HI group were subjected to left common carotid artery ligation,exposed to 80 mL/L oxygen and 920 mL/L nitrogen gas in 37 ℃ closed container for 2.5 h.Rats in 2 groups were killed at 6 hours,24 hours,48 hours,72 hours,5 days after hypoxia respectively.The neuron apoptosis was detected by flow cytometry.The activity of GOD-PX and the contents of SOD and MDA were detected by spectrophotometry,the level of GSK-3? was mensurated by Enzyme-linked immumosorbent assay(ELISA).Results The rates of neuronal apoptosis in HI group were significantly higher than those in normal group at 6,24,48 and 72 hours after hypoxia,respectively(Pa

BACKGROUND:Studies have indicated that melatonin can promote the differentiation of adipose-derived stem cels into neurons, and the effect of melatonin on the osteoblasts form adipose-derived stem cels is rarely reported.
OBJECTIVE:To observe the osteogenic differentiation of adipose-derived stem cels and the effect of melatonin on the bio-viability of differentiated cels.
METHODS:(1) Adipose-derived stem cels were isolated and purified from the inguinal fat of Kunming mice by type I colagenase digestion and differential adhesion method, respectively. Immunohistochemical staining of CD44 was used as a quality control. (2) Osteogenic induction medium was added to induce osteogenic differentiation of passage 2 adipose-derived stem cels. Alkaline phosphatase staining and von Kossa method were combined to evaluate differentiation condition. (3) Melatonin at variable concentrations was added to treat mature osteocytes originated from adipose-derived stem cels and MTT was applied to determine their viability at 24 and 48 hours after culture respectively to find out optimal condition of melatonin treatment. (4) Melatonin at the optimal concentration was used to treat differentiated cels and detect alkaline phosphatase activity after 3 days and 6 days respectively.
RESULTS AND CONCLUSION: (1) After seeding for 48 hours, most cels were adherent, and after 4 days, the cels displayed multiple shapes and colonies of different sizes formed. After subculture, cel morphology homogenized as spindle shape. Cels positive for CD44 were brownish yelow, and localized mainly on the cel membrane. (2) Differentiated cels were positive for von Kossa staining and black sediments scattered in the extracelular matrix. Alkaline phosphatase expressed positively, and brown-black particles, appeared within cels. (3) Melatonin supplement improved the viability of differentiated cels; and 1, 10 and 100 μmol/L was observed as the optimal concentrations both at 24 and 48 hours. (4) The intracelular alkaline phosphatatse activity was increased with time in al the groups (P < 0.05). Compared with the blank group, the intracelular alkaline phosphatase activity in Melatonin groups (1, 10 and 100 μmol/L) had nochanges at 3 days, but significantly increased at 6 days (P < 0.05). These findings indicate that melatonin can enhance the proliferation of osteocytes differentiated from adipose-derived stem cels, and improve the activity of intracelular alkaline phosphatase.

Objective To establish the molecular weight distribution of Ganlong capsule by HPSEC the content of the peptide determined by Lowry and Methods The superdex peptide 10/300 GL (10 mm ×300 mm) column was used.The pH=6.0 and phosphate buffer of 0.05 mol/L was used as the mobile phase, containing 0.1 mol/L NaCl.The flow rate was set at 0.7 mL/min;The column temperature was 25℃;The detection wavelength was 214 nm.Results The content of the peptide ranged from 0.08 mg to 0.4 mg ( r =0.9996 ) .The RSDs of measurement precision of molecular weight and content were 0.08% and 0%(n=6), respectively.The RSDs of the repeatability were 1.3% and 1.1%(n=6);The regression equation of standard material was logMr =5.1455 -0.0871tR, r =0.9983,the relative molecular weight ranged from 2.68 ×102 Da ~5.73 ×103 Da(r =0.9983). Conclusion on The method is simple and rapid for determining the peptide content and the molecular weight distribution of Ganlong capsule.It can be used quality control method for Ganlong capsule.