Autophagy is an important physiological process in eukaryotic cells and is contributive to normal update of cellular components.When the body is in hunger state,autophagy can not only produce amino acid and ATP through degrading proteins to provide conditions for cell survival,but also remove the damaged organelles to maintain the normal functions of cells.The abnormal functions of autophagy are closely related to the formation and development of tumors,and the degree which is too high or too low both can inhibit tumors.It has been found that autophagy is regulated by many kinds of molecules,and the results are different.So in-depth studies on the mechanisms of these autophagy-related molecules will become a starting point for further understanding the autophagy and a sally port for treating tumors.

[Objective] To study the aeroallergens in the patients with allergic rhinitis in Guangzhou.[Methods] Using the German screen allergens quantitative detection system to determinate serum aeroallergen specific IgE and total IgE levels of 625 cases with allergic rhinitis diagnosed in The Third Hospital of SUN Yat-sen University.The samples were stratified on two age groups:the juvenile and the adult.Difference of serum aeroallergen specific IgE of the two groups was investigated.[Results]Aeroallergen specific IgE antibody in the serum of 625 patients with allergic rhinitis were positive.Total IgE in serum was positive in 413 cases,accounting for 66.1%.The positive rate of total IgE was less than that of aeroallergen specific IgE.The positive rate of aeroallergen specific IgE in order were mite mixture (84.32%),cockroach (19.04%),cypress (15.20%),cat-dog epithelia (13.12%),tree mixture (7.20%),ragweed (5.12%),humulus lupulus (1.76%),mugwort (1.76%),mould mixture (1.44%).In the two groups,the positive rate of aeroallergen specific IgE in order were similar.There were significantly higher sensitivity and positive rate of mite mixture in the juvenile group than the adult group (P ＜ 0.01).[Conclusion] The most important aeroallergens are mite mixture,cockroach,cypress,cat-dog epithelia,which could be referenced as Guangzhou patients .The juvenile is more sensitive to mite than the adult.

ABSTRACT]OBJECTIVETo study the compliance to sublingual immunotherapy(SLIT) in patients with allergic rhinitis(AR) in Guangzhou city.METHODSFrom January 2014 to May 2014, 202 patients with AR received SLIT were followed up by telephone. According to age, the patients were divided into group A(age<14 years) and group B(age≥14 years). The compliance to SLIT was analyzed and the reasons of poor compliance were investigated.RESULTSAmong 202 patients, only 93 cases(46.04%) were successfully followed up by telephone, 109 cases(53.96%) were lost to visit. Among the 93 cases of successful follow-up, the good compliance rate was 29.03%(n=27), the poor compliance rate was 70.97%(n=66). compliance to SLIT was not affected by age and gender(P>0.05). Main reasons for poor compliance included poor efficacy (48.48%), insufficient education about SLIT (16.67%), inconvenience (15.15%), and adverse reactions(10.61%).CONCLUSION In Guangzhou city, lost follow-up rate in AR patients receiving SLIT is high. Compliance to SLIT is relatively low and improvements shall be made.

ObjectiveTo determine survival and differentiation of cultured neural stem cells (NSCs) into viable and functional neurons upon transplantation into the brain of MPTP-induced Parkinson's Disease (PD) mode micel.MethodsMice model of PD was established with intraperitoneal (i.p.) injections of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP,40mg/kg) twice interspersed by 16 hr. Tissues isolated from an embryonic rat midbrain were cultured in clonal density. After labeled with 5-bromo-2′-deoxyuridine (BrdU), the NSCs were transplanted into the uni- or bilateral striatum of PD mice. Tyrosine hydroxylase (TH) immunofluorescence was used to evaluate the toxicity of MPTP on the neural cells in the substantia nigra. Immunohistochemistry and laser confocal were used to detect the survival and differentiation of transplanted NSCs.ResultsTH positive neural cells were significantly reduced in the substantia nigra after MPTP injection. Immunohistology showed that the uni-or bilateral transplanted NSCs could survive in the brains of PD model mice.Some transplanted NSCs could properly differentiate into targeted TH positive neural cells in vivo.ConclusionsThe transplanted multipotent NSCs could survive, differentiate into functional dopamine neurons in the brains of PD model mice.

ObjectiveTo observe the changes of neurogenesis in the hippocampus of Alzheimer's model mice and the effect of traditional Chinese medicine Nao Fu Cong. MethodsAlzheimer's type dementia of mice was induced by Aβ25-35 icv.Space learning and memorial ability were tested with Morris Water Maze.The activity of NMDA and M receptor were measured with radio-ligand of MK-801 and QNB.Neurogenesis was observed with the BrdU immunohitochemistry.ResultsSpace learning and memorial ability significantly decreased(P<0.05),MK-801 binding increased and QNB binding decreased (P<0.001),BrdU positive cells decreased in hippocampus(P<0.05).After given drugs for 2 weeks,the mentioned changes were improved significantly(P<0.05,P<0.001,P<0.05).ConclusionsThe toxic effect of Aβ25-35 was involved with the inhibitory action of neurogenesis.Promoting the neurogenesis may be one of the mechanism of traditional Chinese medicine to treat the neurodegenerative diseases.

[Objective] To investigate the effects of airway dysbacteriosis on the development of murine atlergic airway diseases (AAD).[Methods] Female C57BL/6 mice were neubulized with Vancomycin for 10 days and then were sacrificed.The bacterial population in bronchoalveolar lavage fluid (BALF) were evaluated using 16S rRNA high-throughput sequencing technology,exploriug the method of establishing an airway dysbacteriosis mouse model.After the mouse model was established successfully,airway dysbacteriosis mouse models were established by the same method,and based on that,the mice were sensitized and challenged with ovalbumin (OVA) to induce airway allergic inflammation.The frequency of nasal rubbing behaviors per mice was counted;the total cell number and eosinophil relative abundance in BALF were evaluated;the lung tissue inflammation and goblet cell metaplasia were assessed according to histopathological features;and the IgE level in serum,IFN-γ,IL-4 and IL-5 levels in BALF,and IL-33 levels in serum,BALF and intestine tissue were measured by ELISA.[Results] Nebulization of Vancomycin increased Bradyrhizobium,Sphingopyxis,Cupriavidus,Pelomonas,and decreased Akkermansia and Prevotella_6 in airway,inducing significant airway dysbacteriosis.Using the animal model,further study found that airway dysbacteriosis exacerbated OVA-induced airway allergic inflammation,including increased nasal rubbing frequency,higher serun IgE level,more total cell count especially eosinophil infiltration,more serious lung tissue inflammation and goblet cell metaplasia.Additionally,compared to OVA group,mice in Dysbacteriosis and OVA group had significantly increased level of Th2 cytokine IL-4 and IL-5,and significantly decreased Thl cytokine IFN-γin BALF,which revealed that mice in Dysbacteriosis and OVA group had mote remarkable Thl/Th2 imbalance.Furthermore,IL-33 level showed a significant increase in BALF,but didn't change in serum or intestine tissue in Dysbacteriosis and OVA group compared to OVA group.Indicating that airway dysbacteriosis may only affect the local production of IL-33.[Conclusions] An airway dysbacteriosis mouse model was established by Vancomycin nebulization successfully.Airway dysbacteriosis may activate innate lymphoid cells (ILC) and Th2 cell by inducing local IL-33 secreting,which leads to the imbalance of Th1/Th2,and in turn promotes the development of AAD.

Objective Folate receptors ( FRs) , overexpressed on the surface of a variety of tumor cells , are potential targets for tumor targeting therapy .This study aimed to prepare an FR-mediated drug nanocarrier with folate conjugated pullulan acetate ( FPA) to target chemotherapeutic agents to FR-overexpressed tumor cells and investigate its tumor-suppressing effect in vitro. Methods The cytotoxicity of epirubicin-loaded FPA nanoparticles ( FPA/EPI NP) against HepG2 and Hela cells was evaluated by MTS assay.The HepG2 and Hela cells were divided into five groups to be treated with NPs (NP control), chlorpromazine, chloro-quine, amiloride, and folate, respectively, followed by detection of the fluorescence intensity by flow cytometry . Results FPA/EPI NP was successfully formulated into NPs , with the mean particle size of (268.5 ±12.0) nm, by dialysis with an almost spherical shape . The drug-loading rate and entrapment rate of FPA/EPI NP were (6.45 ±1.04) and (72.45 ±11.50) %, respectively.The survival rates of the HepG2 and Hela cells were both >95%after 24 hours of incubation with FPA NP at 5, 40, 200, 400 and 1000μg/mL and 90.0%after 72 hours.The survival rates of the HepG2 cells treated with 5, 40, 200, 400 and 1000μg/mL FPA/EPI NP for 24 hours were (92.3 ±5.2), (70.4 ±4.6), (50.0 ±4.0), (41.1 ±4.1) and (27.0 ±3.6) %, respectively.Compared with the NP control group, the Hela cells of the chlorpromazine , amiloride and folate groups showed a significantly lower rate of NP uptake (P<0.05), and so did the HepG2 cells pretreated with chlorpromazine or amilo-ride (P<0.05).At 72 hours, the half maximal inhibitory concentrations (IC50) of FPA/EPI NP against HepG2 and Hela cells were 168 and 105μg/mL, respectively . Conclusion Clathrin-mediated endocytosis and macropinocytosis are involved in the internaliza-tion of FPA/EPI NP in HepG2 cells, while clathrin-and FR-mediated endocytosis in that of Hela cells .FPA NP may serve as a new drug carrier for tumor-targeted therapy .

Objective To study the conditions and methods of hydrodynamics-based transgene into rat regenerating liver in vivo. Methods The solution with concentration 30mg/L gene-containing plasmid was injected into rat tail veins at a speed of 2ml/s, then partial hepatectomy (PH) was performed at different times before/after injection, finally the rat (g) and regenerating liver (g) were weighed, and the liver coefficient (Lc) was calculated. Out of 15 groups which are Lc±Lc*0%, *5%, *10%, *15%, *20%, *25%, *30%, *35%, the most suitable group was chosen as correction coefficient to calculate the most appropriate volume of plasmid solution which was injected into the regenerating liver at different recovery times, and at the same time, right lobe of liver was gathered to make frozen section, then observe and quantify the positive green fluorescent protein (GFP) rate at 488 nm excitation wavelength. Results Injection of either physiological saline or empty plasmid has no significant difference compared with control (only PH performance). The appropriate time of hydrodynamics-based transgene is more than 12 hours before PH or anytime after PH. The solution volume of hydrodynamics-based transgene into liver regenerating rat after PH is rat weight (g) ×9%×1/3×corresponding correction coefficient (Trc). Both vector and target gene have effect on the time and abundance of gene expression. Conclusion Hydrodynamics-based transgene can effectively be applied to gene transfection in rat regenerating liver.

Objective To investigate the biological effects of 12C heavy ions with different LETs,and to construct the dose-effect curves of chromosomal aberrations and micronucleus.Methods Human peripheral blood lymphocytes were irradiated with 0.5 - 5.0 Gy of 60Co γ-rays,29 and 148 keV/μm 12C with a dose rate of 0.5 Gy/min,respectively.The colchicine method and cytokinesis-blocking method were applied to measure chromosome aberrations and micronuclei formation,respectively. Results The chromosomal aberrations induced by 29 and 148 keV/μm of 12C ion had a linear dose response,while the dose-response of micronucleus induction followed to a linear-quadratic model.Chromosome aberration and micronucleus frequency increased with the LET of 12C ions. However, the micronucleus frequency increased slowly when the radiation doses exceeded 3.0 Gy..Chromosomal aberrations induced by 148 keV/μm 12C ions increased significantly when the time of colchicine treatment was prolonged.Conclusions Chromosome aberration and micronucleus induced by 12C ions increased with the LET,and the micronucleus frequency increased with dose less than 3 Gy.The biological effects of 12C ions were stronger than those of low LET irradiation of γ-rays.

Objective]In order to study the impact of adenoid hypertrophy(AH)with allergic rhinitis(AR)in the otitis media with effusion(OME)in children,as well as to discuss the risk factors on the inducement of OME in patients with AH.[Methods]The clinical materials were collected and analyzed from 205 children with AH who admitted for surgical treatment between 2013 and 2015, including medical history and signs,acoustic immittance measurement,allergy screen as well as blood routine,and to evaluate the situation of OME and AR in patients with AH. All the data were analyzed by SPSS 20.0.[Results]Among the 205 AH children,66 cases(32.20%)were accompanying with OME. The AH patients aged 3-5 years had the highest incidence of OME,which decreased with age(P=0.018). It is significant on the prevalence of the OME in patients with AH between the two groups with and without AR (P = 0.010). In the logistic model investigating the risk factors of inducing OME among patients with AH,the third-degree AH, Type-three and the accompanying with AR were significant. The third-degree and third-type AH children were 2.729 and 6.390-folds higher than others respectively (P = 0.047,P = 0.001). The incidence of OME appeared to be 1.212-folds higher among patients with AR(P = 0.010).[Conclusion]Mechanical obstruction and AR played a superimposed role in the inducement of OME among patients with AH,which was effected by multiple factors. We should pay attention to the“lateral respiratory”allergic diseases on eustachian tube and middle ear.