To investigate the effect of polydatin（PD） on the airway inflammation in the ovalbumin（OVA）-induced asthmatic mice，and to clarify its possible mechanism.

A total of 50 female BALB / c mice were randomly divided into control group， OVA model group， low，middle and high doses of PD groups；there were 10 mice in each group.The experimental asthmatic mouse models were established by OVA induction and stimulation. The mice were sensitized by intraperitoneal injection the mixture of OVA and Al （OH）₃ on the 1st， 7th and 14th days of experiment. From the 21st day， the mice in different doses of PD groups were injected with 15，30，and 45 mg·kg^－1 PD， while the mice in control group and OVA group were given normal saline. HE and PAS staining were used to observe the pathomophology of lung tissue of the mice in various groups； Diff-Quick method was used to detect the total number of cells and the number of eosinophils， neutrophils and lymphocytes in bronchoalveolar lavage fluid（BALF）； the levels of IL-4， IL-5， and IL-13 in BALF of the mice in various groups were detected by ELISA method； immunohistochemistry was used to observe the expressions of SIRT1 and NF-κB p65 in lung tissue of the mice in various groups； Western blotting method was used to detect the expression levels of SIRT1 protein and acetylized NF-κB p65 protein in lung tissue of the mice in various groups.

Compared with control group，the airway wall of the mice in OVA group was thickened， a large number of inflammatory cells infiltrated around the airway and the goblet cells were proliferated significantly.Compared with OVA group，the findings mentioned above in different doses of PD groups were improved. Compared with control group， the total number of cells，the number of eosinophils， neutrophils，and lymphocytes and the levels of IL-4， IL-5， and IL-13 in BALF of the mice in OVA group were increased （P<0.05）. Compared with OVA group， the total number of cells，the number of eosinophils and neutrophils， and the levels of IL-4， IL-5， and IL-13 in BALF of the mice in middle and high doses of PD groups were decreased （P<0.05）. Compared with control group， the expression levels of SIRT1， NF-κ B p65 and acetylized NF-κB p65 proteins in lung tissue of the mice in OVA group were significantly increased （P<0.05）. Compared with OVA group， the expression levels of SIRT1 protein in lung tissue of the mice in middle and high doses of PD groups were increased （P<0.05）， while the expression levels of NF-κB p65 and acetylized NF-κB p65 proteins in ling tissue of the mice in middle and high doses of PD groups were decreased （P<0.05）.

PD can alleviate the airway inflammation in the asthmatic mice， and its mechanism may be related to the SIRT1/NF-κB pathway.

To explore the protective effect of rutin on the oxidative stress injury of liver cancer HepG2 cells， and to clarify its mechanism.

The liver cancer HepG2 cells in the logarithmic growth phase were divided into control group， tert-butyl hydroperoxide （TBHP） group （400 μmol·L^－1 TBHP）， 0.25， 0.50，and 1.00 μmol·L^－1 rutin groups （0.25， 0.50， 1.00 μmol·L^－1 rutin+400 μmol·L^－1 TBHP）. The vitalities of liver cancer HepG2 cells in various groups and the survival rates of liver cancer HepG2 cells in various groups were detected by MTT assay；the levels of malondialdehyde （MDA） and glutathione （GSH） and activities of superoxide dismutase （SOD） in the liver cancer HepG2 cells in various groups were assessed by colorimetric method； the levels of reactive oxidative species （ROS） in the liver cancer HepG2 cells in various groups were quantified with DCFH-DA immnofluorescence staining. The expression levels of PI3K， p-PI3K， Akt， p-Akt， NF-κB， p-NF-κB，and p53 proteins in the liver cancer HepG2 cells in various groups were mesured by Western blotting method.

The MTT results showed that when the concentrations of rutin were less than or equal to 1.00 μmol·L^－1， the vitality of liver cancer HepG2 cells was higher than 90%， and the rutin had no toxic effect； when the concentration of rutin was higher than 1.00 μmol·L^－1， the vitality of the liver cancer HepG2 cells was lower than 90%，and the rutin had toxic effect. Compared with control group， the survival rate of the HepG2 cells in TBHP group was significantly decreased （P<0.01）； compared with TBHP group， the survival rates of the HepG2 cells in different concertrations of rutin groups were significantly increased （P<0.01）. Compared with control group， the level of MDA in the HepG2 cells in TBHP group was significantly decreased （P<0.01）， the level of GSH and the activity of SOD were significantly increased （P<0.01）； compared with TBHP group， the levels of MDA in the HepG2 cells in different concentrations of rutin groups were significantly increased（P<0.01）， the levels of GSH and the activities of SOD in the HepG2 cells in different concertrations of rutin groups were significantly decreased （P<0.01）. The results of DCFH-DA immunofluorescence staining showed that the ROS level in the HepG2 cells in TBHP group was higher than that in control group； compared with TBHP group， the ROS levels in the HepG2 cells in different concentrations of rutin groups were gradually decreased（P<0.05）. The Western blotting results showed that compared with control group， the expression levels of PI3K， p-PI3K， Akt， and p-Akt proteins in the HepG2 cells in TBHP group were decreased （P<0.05）， and the expressions levels of NF-κB， p-NF-κB，and p53 proteins were increased （P<0.05）； compared with TBHP group， the expressions levels of PI3K， p-PI3K， Akt， and p-Akt proteins in the HepG2 cells in different concentrations of rutin groups were increased （P<0.05）， and the expressions levels of NF-κB， p-NF-κB， and p53 proteins were decreased （P<0.05）.

Rutin has the protective effect on TBHP-induced HepG2 cell damage， and its mechanism may be related to the regulation of PI3K/Akt and NF-κB pathways.

To observe the effect of acupuncture combined with medicine on the autophagy in hippocampus tissue of the mice with ApoE gene knockout， and to explore the mechanism of acupuncture combined with medicine in regulating the autophagy through the phosphatidylinosital-3-kinase（PI3K）/protein kinase B（AKT）-mamalian target of repamycin（mTOR） signaling pathway.

Fifty 8-week-old ApoE gene knockout mice were fed with high-fat diet， and after 16 weeks， the ApoE gene knockout mice were randomly divided into model group， low dose of traditional Chinese medicine（TCM） group， high dose of TCM group， acupuncture group， acupuncture+ TCM group；there were 10 rats in each group.Another 10 C57BL/6L mice with the same age fed with normal diet were regarded as control group. From the 17th week， the mice in each group except blank group were given intervention for 8 weeks. After the intervention， the escape latency time of the mice in various groups was observed by Water Maze experiment； the ultrastructures of hippocampus tissue of the mice in various groups were observed by transmission electron microscope； Western blotting and immunohistochemistry methods were used to detect the expression levels of PI3K，AKT and mTOR proteins in hippocampus tissue of the mice in various groups.

The Water Maze results showed that the escape latency time of mice in model group was significantly longer than that in control group （P<0.05）， and the escape latency of the mice in acupuncture group was significantly shorter than that in model group （P<0.05）. The transmission electron microscope observation results showed that the neurons in hippocampal region of the mice in model group appeared deformed， irregular or atrophied， and the nuclear membrane was rough and incomplete， the double-layer structure was fuzzy， and the intracellular phagocytosis was reduced； the neurons in the hippocampal region of the mice in acupuncture group still had some irregular deformation and the arophy and autophagic vesicles could be seen in the cytoplasm. The Western blotting and immunohistochemistry results showed that compared with control group， the expression levels of PI3K， AKT， and mTOR proteins in hippocampus tissue of the mice in model group were significantly increased （P<0.05）； compared with model group， the expression levels of PI3K， AKT， and mTOR proteins in hippocampus tissue of the mice in high dose of TCM group， acupuncture group，and acupuncture+ TCM group were decreased（P<0.05），especially in acupuncture+TCM group （P<0.05）.

The combination of acupuncture and medicine can improve the cognitive function of ApoE gene knockout mice， and its mechanism may be achieved by inhibiting the excessive activation of the PI3K/AKT/mTOR signaling pathway and promoting the autophagy in hippocampal cells.

To investigate the intervention effects of single acupoint and acupoint compatibility on the gastric mucosal injury index， the morphology of gastric mucosa and the expression of epidermal growth factor receptor （EGFR ）protein in gastric tissue of the rats with gastric ulcer， and to clarify the effect of single acupoint and acupoint compatibility and the mechanism of acupuncture for treatment of gastric ulcer in the rats.

A total of 100 SD rats were randomly divided into blank group， model group， Zusanli group， Zhongwan group， and acupoint compatibility group，and there were 20 rats in each group. Except for blank group， the rats in the other groups were subjected to restraint-water immersion stress method to establish the stress gastric ulcer rat models.After the models were successfully established， no intervention was given to the rats in blank group and model group，and the rats in Zhongwan group， Zusanli group，and acupoint compatibility group were respectively treated with Zhongwan， Zusanli， and Zusanli+Zhongwan electroacupuncture treatment. After 10 d， the gastric ulcer of the rats in various groups was observed， the gastric mucosal injury indexes of the rats in various groups were detected， the pathomorphology of gastric tissue of the rats in various groups was observed by HE staining， and the expression levels of EGFR protein in gastric mucosa tissue of the rats in various groups was detected by immunohistochemistry.

The gastric ulcer index of the rats in model group was higher than that in blank group （P<0.05）， and the gastric ulcer indexes of the rats in Zhongwan group， Zusanli group and acupoint compatibility group were lower than that in model group （P<0.05）. The HE staining results showed that the gastric mucosa of the rats in model group was destroyed，and the gastric mucosa of the rats in Zusanli group， Zhongwan group， and acupoint compatibility group had varying degrees of repair and healing，especially in acupoint compatibility group. The expression level of EGFR protein in gastric mucosa tissue of the rats in model group was higher than that in blank group （P<0.05）；compared with Zhongwan group，the expression level of EGFR protein in gastric mucosa tissue of the rats in Zusanli group was increased （P<0.05），and the expression level of EGFR protein in gastric mucosa tissue of the rats in acupoint compatibility group was increased （P<0.05）； compared with Zusanli group， the expression level of EGFR protein in gastric mucosa tissue of the rats in acupoint compatibility group was increased （P<0.05）.

Electroacupuncture has therapeutic effect on the gastric ulcer by promoting the expression of EGFR， and its mechanism may be related to that EGFR can control the proliferation， differentiation and survival of the gastric mucosal cells.

To investigate the effect of M1 macrophages on the immune status of the model mice with porphyromonas gingival （P.gingivalis ）-induced chronic periodontitis， and to clarify the role of M1 macrophages in chronic periodontitis.

Ten 7-week-old female C57BL/6 mice were randomly divided into control group（n=5）and chronic periodontitis group（n=5）.The chronic ?periodontitis?models of mice were induced? by? oral? infection? with P.gingivalis. The resorption of alveolar bone of the mice in two groups was estimated by measuring the distance from the cemento-enamel junction（CEJ） to the alveolar bone crest（ABC）；HE staining was performed to examine the infiltration of inflammatory cells， attachment loss of connective tissue and resorption of alveolar bone in periodontal tissue of the mice in two groups.? flow cytometry was used to detect the percentages and number of F4/80^＋iNOS^＋ M1 macrophages in gingival tissue of the mice in two groups； real-time quantitative PCR method was used to detect the expression levels of iNOS mRNA in gingival tissue of the mice in two groups.

Compared with control group， the distance from CEJ to ABC of the mice in chronic periodontitis group was increased significantly（P<0.01）； the gingival junction epithelium attached to the root side of CEJ， and the absorption of ABC became flat， the osteoclasts and bone resorption lacunae of the mice in chronic periodontitis group were found.The number and percentage of M1 macrophages in gingival tissue of the mice in chronic periodontitis group were significantly higher than those in control group（P<0.01）.Compared with control group，the expression level of iNOS mRNA in gingival tissue of the mice in chronic periodontitis group was significantly increased（P<0.01）.

In the mouse model of chronic periodontitis， the immune response mediated by M1 macrophages is enhanced， and M1 macrophages are involved in the inflammatory response and tissue damage of chronic periodontiti.

To explore the expression differences of proteins in the neural stem cells （NSCs） in hypothalamic tissue and hippocampal tissue in the C57BL/6J mice by proteomic analysis and clarify the reasons for different protein expression profiles of NSCs from different sources （regions）， and to provide the reference for the basic research and transplantation treatment of NSCs in hypothalamic and hippocampal tissues.

The primary NSCs were isolated from the newborn C57BL/6J mice within 24 h， and were expanded and purified by passage. The morphology of cells was observed； the expressions of Nestin and Sox2 were detected by immunofluorescence method；the differential proteins of NSCs in hypothalamic and hippocampal tissues were analyzed by tandem mass tag （TMT） proteomics， and the obtained differential proteins were analyzed by bioinformatics analysis.

The morphology detection results showed that the NSCs in hypothalamic and hippocampal tissues presented the typical neurosphere morphology with opacity and high refractive index. The immunofluorescence detection results showed that the positive expression rates of Nestin and Sox2 in the NSCs in hypothalamic and hippocampal tissues were increased with the increase of culture generation， and it reached over 98% in the 4th generation， but the difference in the NSCs between hypothalamic and hippocapal tissues was not statistically significant（P>0.05）. The proteomics analysis results showed that compared with the hippocampal tissue，the expression levels of 6 kinds of proteins in the NSCs in hypothalamic tissue were significantly increased（P<0.05 or P<0.01）， while the expression levels of 4 kinds of proteins were significantly decreased（P<0.05 or P<0.01）. The gene ontology （GO） analysis results revealed that the differential proteins were mainly involved in 8 biological processes such as negative regulation of dopamine uptake involved in synaptic transmission， 6 kinds of molecular functions such as rRNA （guanine） methyltransferase activity and so on， and 6 cellular components such as integral component of Golgi membrane and so on. The Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway analysis results revealed that the differential proteins were mainly involved in 7 signaling pathways such as SNARE interactions in the vesicular transport and so on.

The primary NSCs in hypothalamic and hippocampal tissues from C57BL/6J mice which are cultured in vitro have no differences in morphology and the expressions of NSCs markers. The expressions of certain specific proteins are different，the differentially expressed proteins mainly play a role in synaptic transmission， vesicle transport， and metabolism and may play an important role in the orientation differentiation and functional specificity of NSCs in different regions.

To construct the GLI1 eukaryotic expression vector，and to discuss the effect of over-expression of GLI1 on the proliferation of lung cancer PC9 cells.

The full length sequence of GLI1 gene was amplified by PCR method and it was cloned into the pcDNA3.1 eukaryotic expression vector. The PC9 cells were transfected with the recombinant vector pcDNA3.1-GLI1 （recombination vector group） and the control vector pcDNA3.1 （empty vector group）. The construction of GLI1 over-expression vector was detected by enzymatic digestion identification method. The cells stably expressed with GLI1 and empty vector were established by G418 screening. The expression levels of GLI1 mRNA in the PC9 cells in two groups were detected by RT-PCR method； the expression levels of GLI1 protein in the PC9 cells in two groups were detected by Western blotting method；the proliferation activities of the PC9 cells in two groups were detected by CCK-8 method； the clone formation number of the PC9 cells in two groups was detected by cell clonal formation assay.

The GLI1 gene fragment was specifically amplified successfully； the target genes and vector fragments were obtained by double enzyme digestion of the GLI1 eukaryotic expression vectors. The RT-PCR results showed that the expression level of GLI1 mRNA in the PC9 cells in recombination vector group was significantly higher than that in empty vector group （P<0.01）. The Western blotting results showed that the expression level of GLI1 protein in the PC9 cells in recombination vector group was significantly higher than that in empty vector group （P<0.05）. The CCK-8 results showed that the proliferation activities of the PC9 cells in recombination group were significantly higher than those in empty vector group at 72 and 96 h after culture （P<0.01）. The clonal formation assay results showed that the clone formation number of the PC9 cells in recombination group was significantly more than that in empty vector group at 10 d of culture （P<0.01）.

The GLI1 eukaryotic over-expression vector and the PC9 cells stably over-expressed with GLI1 are established successfully. Over-expression of GLI1 can promote the proliferation ability of lung cancer PC9 cells.

To investigate the regulatory effect of forkhead box O1（FOXO1） on the alveolar epithelial sodium channel （ENac） in the lipopolysaccharide （LPS）-induced acute lung injury mice， and to illustrate the mechanism.

Twenty C57BL/6J mice were randomly divided into control group and LPS group， and there were 10 mice in each group. After treatment， the mouse samples were obtained. HE staining was used to observe the pathomorphology of lung tissue of the mice in two groups， RT-PCR method and Western blotting method were used to detect the expression levels of α-subunit ENaC（αENaC） mRNA and pFOXO1 （Ser256） protein in lung tissue of the mice in two groups.The alveolar epithelial A549 cells were cultured， and then infected with ADV-FOXO1 or shRNA-FOXO1， respectively.RT-PCR method was used to determine the expression of αENaC mRNA. The A549 cells were divided into control group， insulin group， insulin+PI3K inhibitor group， and insulin+AKT inhibitor group. After corresponding intervention， Western blotting method was used to detect the expression levels of pFOXO1 （Ser256） protein，RT-PCR method was used to determine the expression levels of αENaC mRNA， and immunofluorescence method was used to detect the intracellular localization of FOXO1.

Compared with control group， the lung injury score of mice in LPS group was increased significantly （P<0.05）， the expression levels of pFOXO1 （Ser256） and αENaC proteins in the lung tissue were significantly decreased（P<0.05） and they showed positively correlated relationship（r=0.703， P<0.05）.Compared with control group； the expression level of αENaC mRNA in A549 cells in over-expression FOXO1 group was significantly decreased （P<0.05），and the expression level of αENaC mRNA in silencing FOXO1 group was significantly increased（P<0.05）； compared with control group， the expression levels of pFOXO1 protein and αENaC mRNA in the A549 cells in insulin group were increased significantly （P<0.05）， and FOXO1 was mainly located in the cytoplasm. The expression levels of αENaC mRNA and pFOXO1 protein in A549 cells in insulin+PI3K inhibitor group and insulin+AKT inhibitor group were lower than those in insulin group （P<0.05）.

After inactiviation of FOXO1 phosphorylation，FOXO1 can translocate from nucleus to cytoplasm to weaken the inhibitory effect of αENaC and upregulate the expression of αENaC.

To explore the effect of astragalus polysaccharide （APS） on the cisplatin（DDP） resistance of human lung cancer A549/DDP cells， and to clarify its possible mechanism.

The A549/DDP cells at logarithmic growth phase were divided into control group （without drug intervention）， APS groups （ given 25， 50， 100， 200， and 400 mg·L^－1 APS）， DDP groups （given 2， 4， 8， 16， and 32 mg·L^－1 DDP） and APS+DDP groups （given 100 mg·L^－1 APS and 2， 4， 8， 16， 32 mg·L^－1 DDP）. After the cells were treated for 24 h， the inhibitory rates of proliferation of the A549/DDP cells in various groups were detected by CCK-8 method and the half inhibitory concentration （IC₅₀） was caluculated. The A549/DDP cells were divided into control group （without drug intervention）， APS group （given 100 mg·L^－1 APS）， DDP group （given 11.46 mg·L^－1 DDP） and APS+DDP group （given 100 mg·L^－1 APS and 11.46 mg·L^－1 DDP）， the cells in various groups were treated with drugs for 24 h. The migration of A549/DDP cells in various groups was detected by Transwell chamber assay； the apoptotic rates and mitochondrial membrane potentials of the A549/DDP cells in various groups were detected by flow cytometry， and the level expression levels of B-cell lymphoma 2 （Bcl-2）， Bcl-2-related X protein （Bax），P-glycoprotein （P-gp），phosphatidylinositol-3 kinase （PI3K）， phosphorylated phosphatidylinositol-3 kinase （p-PI3K）， protein kinase B （AKT） and phosphorylated protein kinase B （p-AKT） proteins in the A549 / DDP cells in various groups were detected by Western blotting method.

The CCK-8 results showed that compared with DDP group， the IC₅₀ of DDP in APS+DDP group was decreased. The Transwell chamber assay results showed that compared with control group， the migration number of A549 / DDP cells in APS group was not significantly changed； compared with DDP group， the migration number of A549 / DDP cells in APS+ DDP group was decreased.The results of flow cytometry showed that compared with control group， the apoptotic rate and mitochondrial membrane potential of A549 / DDP cells in APS group had no significant changes （P>0.05）， and the apoptotic rate and mitochondrial membrane potential of the A549/DDP cells in DDP group were significantly increased （P<0.05）； compared with DDP group， the apoptotic rate and mitochondrial membrane potential of A549 / DDP cells in APS+DDP group were significantly increased （P<0.05）；the Western blotting results showed that compared with control group， the expression levels of Bcl-2 and P-gp proteins in the A549/DDP cells in APS group was significantly decreased （P<0.05）， the ratios of p-PI3K/PI3K and p-AKT/AKT were decreased（P<0.05）， and there was no significant difference in the expression level of Bax protein （P>0.05）；compared with APS group， the expression levels of Bcl-2 and P-gp proteins in the A549 / DDP cells in DDP group were significantly decreased （P<0.05），the ratios of p-PI3K/PI3K and p-AKT/AKT were decreased（P<0.05）， and the expression level of Bax protein was significantly increased （P<0.05）；compared with DDP group， the expression levels of Bcl-2 and P-gp proteins in the A549 / DDP cells in DDP+APS group were significantly decreased （P<0.05）， the ratios of p-PI3K/PI3K and p-AKT/AKT were decreased（P<0.05），and the expression level of Bax protein was significantly increased （P<0.05）.

APS can reverse the drug resistance of A549/DDP cells，and its mechanism may be related to blocking the PI3K/AKT signal pathway and inducing the mitochondrial damage.

To investigate the damage of bone marrow hematopoietic stem/progenitor cells （HSCs/HPCs） and the changes of CD47 expression levels in the HSCs/HPCs of the mice after X-ray irradiation，and to clarify the role of CD47 in irradiation-induced injury of HSCs/HPCs.

The C57BL/6 mice were divided into nonirradation group and irradation group， and the C57BL/6 mice in irradation group were divided into 18 h， 5 d， 11 d， and 6 weeks after irradiation subgroups，and there were 5 mice in each group. The mice in irradation groups were given by an X-ray irradiator with a dose of 3.0 or 9.0 Gy at one time. The mice were sacrificed at different time points after irradiation.The percentages of HSCs to HPCs in bone marrow of the mice in various groups were detected by flow cytometry， and the expression levels of CD47 in HSCs/HPCs of the mice in various groups were detected at different time points after irradiation.

The level of CD47 expression in HPCs of the mice in non-irradation group was more than 2-fold higher than the mature HSCs in bone marrow （P<0.05）. Compared with non-irradation group， there were no statistical differences in the CD47 expression levels in HPCs of the mice in irradation groups at 18 h， 5 d， and 11 d after irradation （P>0.05）； the percentages of HPCs in lineage-negative （lin^－） cells and whole bone marrow cells were decreased significantly（P<0.05）， which returned to normal level at 6 weeks after irradiation. Compared with non-irradiation group， the CD47 expression level in HSCs of the mice in 18 h after irradiation group was increased （P<0.01）， and there were no significant differences in the CD47 expression levels in HSCs of the mice in 5 d，11 d and 6 weeks after irradiation groups（P>0.05）； the percentage of HSCs in Lin^－ cells in 18 h after irradiation group was increased （P<0.01）， and the percentage of HSCs cells in whole bone marrow cells in 11 d after irradiation group was decreased （P<0.01）.The percentage of CD47 low expression subgroup was significantly lower than that of CD47 high expression subgroup in the HPCs in 18 h after irradiation group（P<0.01）.The percentage of CD47 low expression subgroup in the HPCs in 5 d，11 d and 6 weeks after irradiation groups were similar to those in CD47 high expression subgroup.

The total body ionizing radiation can cause the bone marrow damage in the mice and the decreasing in the percentage of HSCs to HPCs， and the mechanism may be related to the transient increase of CD47 expression in the HSCs after irradiation.

To discuss the effect of gardenoside on the sleep disorder in the Parkinson’s disease （PD） rats，and to explore its possible mechanism.

Seventy Wistar rats were selected and 6-hydroxydopa（6-OHDA） was used to establish the PD rat models. After 4 weeks， the PD models were identified by rotation test induced by intraperitoneal injection of apomorphine.After the models were successfully constructed， the PD model rats were divided into model group， low dose of gardenoside group and high dose of gardenoside group， and there were 10 rats in each group；another 10 normal rats were used as normal control group.The rats in low and high doses of geniposide groups were given geniposide solution （50 and 100 mg ? kg^－1） by gavage every day. The rats in normal control group and model group were given the same amount of normal saline for 4 weeks.The sleep time of the rats in various groups was recorded using embedded electroencephalogram（EEG）and electromyography（EMG） electrodes，and the levels of dopamine in striatum of the rats in various groups were analyzed by high performance liquid chromatography （HPLC）.

Compared with normal control group， the total sleep time， slow wave sleep time and fast wave sleep time of the rats in model group were significantly shortened （P<0.01）， and the awaken time was significantly prolonged （P<0.05）.Compared with model group， the total sleep time，slow wave sleep time and fast wave sleep time of the rats in low and high doses of gardenoside groups were significantly prolonged （P<0.05）， while the awaken time was significantly shortened （P<0.05）. Compared with model group， the levels of dopamine in striatum of the rats in low and high doses of gardenoside groups were significantly increased （P<0.05）.

Gardenoside can improve the sleep distorder in the PD model rats， and its mechanism may be related to increasing the level of dopamine in striatum.

To investigate the effect of cardamom （CAR） on the proliferation of the cervical cancer HeLa cells， and to clarify its possible mechanism.

The cervical cancer HeLa cells were divided into control group（given no treatment） and different concentrations （10， 20，and 40 μmol?L^－1） of CAR groups（given different concerntrations of CAR）. CCK-8 method was used to detect the viabilities of the cervical cancer HeLa cells in various groups at 24， 48，and 72 h after treatment； the colony formation experiment was used to detect the clone formation number of the cervical cancer HeLa cells in various groups；and the apoptotic morphology of cervical cancer HeLa cells in various groups was observed by Hoechst33258 staining； Western blotting method was used to detect the expression levels of B-cell lymphoma-2（Bcl-2），Bcl-2 associated X protein（Bax），phosphorylated-phosphatidylinositol 3-kinase（p-PI3K），and phosphorylated-protein kinase B（p-AKT）proteins in the HeLa cells in various groups.

Compared with control group， the viabilities of the cervical cancer HeLa cells in different concentrations of CAR groups after treated for different time were decreased （24 h：P=0.011 8； 48 h：P=0.000 4； 72 h：P=0.002 2）；compared with control group， the clone formation number of HeLa cells in different concentrations of CAR groups was significantly decreased （P<0.01）；compared with control group， the apoptotic morphology of HeLa cells in different concentrations of CAR groups showed as nuclear aggregation， shrinkage or fragmentation； compared with control group， the expression levels of Bcl2， p-PI3K（P=0.011 8）， and p-AKT（P=0.034 9） proteins in the cervical cancer HeLa cells in different concentrations of CAR groups were significantly decresed， while the levels of Bax protein were increased， and the ratios of Bcl-2/Bax were decreased（P<0.01）.

CAR can induce the apoptosis and inhibit the proliferation of cervical cancer HeLa cells by regulating the PI3K/AKT signaling pathway.

The pSilencer4.1-B7-H4-shRNA and pSilencer4.1-scrambled shRNA vectors were converted into the HT-29 cells by liposome method as B7-H4 shRNA group and scrambled shRNA group. The expression levels of B7-H4 mRNA in the HT-29 cells in two groups was detected by RT-qPCR method； the expression levels of B7-H4 protein in the HT-29 cells in two groups were detected by Western blotting method； CCK-8 method was performed to detect the proliferation activities of the HT-29 cells in two groups；flow cytometry was used to detect the percentages of HT-29 cells at different cell cycles and the apoptotic rates of the HT-29 cells in two groups； Transwell chamber assay was used to measure the number of migration cells of the HT-29 cells in two groups； the levels of matrix metalloproteinases 2（MMP-2）and matrix metalloproteinases 9（MMP-9）in liquid supernatant of the HT-29 cells in two groups were detected by ELISA assay；the expression amounts of Bcl-2 and Caspase-3 in the HT-29 cells in two groups were detected by Western blotting method；the expression levels of B7-H4 mRNA in HT-29 cells before and after inhibiting the PI3K/Akt/mTOR signal pathway were analyzed by RT-qPCR method；Western blotting method was used to detect the expression levels of B7-H4 protein in the HT-29 cells before and after inhibiting the PI3K/Akt/mTOR signal pathway.

The expression levels of B7-H4 mRNA and protein in the HT-29 cells in B7-H4 shRNA group were significantly lower than those in scrambled shRNA group （P<0.01）. Compared with scrambled shRNA group， the proliferation ability of the HT-29 cells in B7-H4 shRNA group was decreased （P<0.05）， and the percentages of cells in G₀/G₁ phase and S phase had were no statistically significant differences（P>0.05）， the apoptotic rate and the expression level of Caspase-3 protein in the HT-29 cells were significantly increased （P<0.05）， the expression level of Bcl-2 protein was significantly decreased （P<0.05），the number of migrating cells was decreased （P<0.01）， and the levels of MMP-2 and MMP-9 were also significantly decreased （P<0.05）.Compared with control group， the expression levels of B7-H4 mRNA and protein in the HT-29 cells in LY294002 group and rapamycin group were significantly decreased（P<0.05）.

Silencing of B7-H4 expression can significantly inhibit the proliferation， apoptosis and migration of the HT-29 cells， and its mechanism may be related to the activity of PI3K/Akt/ mTOR signaling pathway.

To observe the effect of interleukin-17A-signal transducer and activator of transcription 3-vascular endothelial growth factor（IL-17A-Stat3-VEGF） signaling pathway activation on the migration of prostatic cancer cells， and to explore the relative mechanism.

A total of 73 prostatic samples including 8 cases of normal prostate （NP） tissue （adjacent normal tissue）， 30 cases of benign prostatic hyperplasia （BPH） tissue and 35 cases of prostate cancer tissue. Immunohistochemical staining method was used to detect the expressions of IL-17A， IL-17RA， Stat3 and VEGF in NP， BPH and prostate cancer tissues， and their correlations with the malignancy degrees of prostate cancer were analyzed.ELISA method was used to detect the serum levels of IL-17A， IL-6， VEGF and matrix metallopeptidase-9 （MMP-9） of the subjects in healthy control group and prostate cancer group. The prostatic cancer cells were cultured with IL-17A recombinant protein in vitro， and scratching test and Transwell chamber experiment were used to observe the migration ability of the cells；Western blotting method was used to detect the expression levels of Stat3， p-Stat3 and VEGF in the PC3 cells in various groups.

The immunohistochemical staining results showed that the expression levels of IL-17A and IL-17RA in BPH tissue were significantly increased compared with NP tissue（P<0.05）； the expression levels of IL-17A， IL-17RA， Stat3， and VEGF in prostate cancer tissue were higher than those in NP tissue（P<0.05）. Compared with BPH tissue， the expression levels of IL-17A， IL-17RA， Stat3， and VEGF in prostate cancer tissue were increased （P<0.05）. The ELASA results showed that the serum levels of IL-17A， IL-6，and VEGF in the patients with prostate cancer were significantly higher than those in healthy controls（P<0.05）. In the prostate cancer tissue， the expression levels of IL-17A， IL-17RA， Stat3， and VEGF were significantly increased with the increasing of Gleason score （P<0.05）. After the PC3 cells were co-cultured with IL-17A recombinant protein for 24 h， the number of migration cells in 50 and 100 μg · L^－1 IL-17A groups was significantly increased compared with blank control group （0 μg · L^－1 IL-17A） （P<0.05）；the expression levels of Stat3，p-Stat3，and VEGF in the PC3 cells in 50 and 100 μg·L^－1 IL-17A groups were significantly increased compared 0 and 20 μg·L^－1 IL-17A groups （P <0.05）.

IL-17A can obviously promote the migration of prostatic cancer cells， and its mechanism may be related with the activation of IL-17A-stat3-VEGF signaling pathway.

To observe the effects of Astragalus Injection on the proliferation of human keratinocytes（KC） and the expression of adhesion factor β-catenin， and to explore the possible mechanism of Astragalus Injection in the treatment of psoriasis.

The HaCaT cells were collected. In the proliferation function experiment， the HaCaT cells were divided into experimental groups （added with different concentrations of Astragalus Injection） and control group （without drugs）， at the same time， blank group （without drugs and cells） was set up. The effective concentration of Astragalus Injection was determined by cell counting method.The HaCaT cells in experimental groups were incubated for 24， 48 and 72 h after adding different concentrations of Astragalus Injection with two times gradient dilution， and the proliferation rates of HaCaT cells in various groups were detected by CCK-8 method. In the adhesion function experiment， the HaCaT cells in experimental group were treated with 1.0 g·L^－1 Astragalus Injection and the HaCaT cells in control group were not treated with Astragalus Injection. After cultured for 24 and 48 h， the expression levels of β- catenin mRNA in the HaCaT cells in two groups were detected by qPCR method； after cultured for 48 h， the expression levels of β-catenin protein in the HaCaT cells in two groups were detected by Western blotting method.

The proliferation function test results showed that compared with 0，0.01， and 0.10 g·L^－1 Astragalus Injection groups，the proliferation rates of the HaCaT cells in 1.0 g·L^－1 Astragalus Injection group were decreased after treated for 24 and 48 h （P<0.01）. The HaCaT cells were incubated with 2 times concentration gradient dilution of Astragalus Injection （ 0， 0.5， 1.0， 2.0， 4.0， 8.0， 16.0 g·L^－1）for 24， 48， and 72 h， compared with control group， the proliferation rates of HaCaT cells in 1.0 g·L^－1 Astragalus Injection group were decreased after treated for 24 and 48 h （P<0.05）， the proliferation rate of HaCaT cells was increased after treated for 72 h（P<0.05）. In the adhesion function experiment， compared with 0 g·L^－1 Astragalus Injection group， there was no significant difference in the expression level of β-catenin mRNA in the HaCaT cells in 1.0 g·L^－1 Astragalus Injection group after treated for 24 h （P>0.05）；after treated for 48 h， the expression level of β-catenin protein in the HaCaT cells in 1.0 g·L^－1 Astragalus Injection group was decreased significantly （P<0.01）.

The mechanism of Astragalus injection in the treatment of psoriasis may be related to the inhibition of proliferation of human KC and the expression of adhesion factor β-catenin. As a single factor， Astragalus Injection could inhibit the proliferation of HaCaT cells and reduce the expression of β-catenin protein，and the inhibitory effect may decrease with the prolongation of time.

To investigate the pharmacodynamic effects of nasal inhalation of microRNA-124（miRNA-124） nanoparticles on the durg distribution in brain tissue， neurological function score， water content in brain tissue and levels of serum inflammatory factors in the ischemic stroke model rats， and to elucidate its neuroprotective effect on the rats with ischemia stroke.

The target RVG29 was coupled to the poly（ethylene glycol）-poly（lactic-co-glycolic acid）（PEG-PLGA） nanoparticles， the particle size and encapsulation efficiency of the prepared nanoparticles were measured.The PEG-PLGA nanoparticles modified with neurotropic virus-derived peptide RVG29 labeled near-infrared dye 1，1'-dioctadecyl-3，3，3，3-tetramethylindole three-carbon cyanine iodide （DiR）were given to the rats， and the durg biodistribution at different time points after nasal inhalation was measured. The rat model of cerebral ischemia-reperfusion was established by the suture method， and 90 rats were randomly divided into sham operation group， model group， Fasudil group， miRNA-124 nasal inhalation group， PEG-PLGA/miRNA-124 nasal inhalation group， and RVG29-PEG-PLGA/miRNA-124 nasal inhalation group， and there were 6 rats in each group. The neurological function scores and water contents in brain tissue of the rats in various groups were evaluated； the levels of interleukin-4（IL-4）， interleukin-6（IL-6）， tumor necrosis factor-α（TNF-α）， and interleukin-1β （IL-1β）in peripheral serum of the rats in various groups were detected by ELISA method.

The average particle size of RVG29-PEG-PLGA nanoparticles was 204 nm， and the encapsulation efficiency was 28%. The rats in sham operation group had no neurological dysfunction. Compared with sham operation group， the neurological function score， water content in brain tissue， and levels of serum IL-4， IL-6， TNF-α， and IL-1β of the rats in model group were increased （P<0.05）. Compared with model group， the neurological function scores， water contents in brain tissue， and levels of serum IL-4， IL-6， TNF-α， and IL-1β of the rats in Fasudil group， miR-124 nasal inhalation group， PEG-PLGA/miRNA-124 nasal inhalation group， and RVG29-PEG-PLGA/miRNA-124 nasal inhalation group were decreased （P<0.05）. Compared with Fasudil group， the neurological function score， water content in brain tissue， and levels of serum IL-4， IL-6， TNF-α， and IL-1β of the rats in RVG29-PEG-PLGA/miRNA-124 nasal inhalation group were decreased （P<0.05）.

The RVG29-PEG-PLGA nanoparticles have a good ability of nasal inhalation into the brain. Intervention of nanoparticle-carrying miRNA-124 of the rats with ischemic stroke can improve the neurological dysfunction， reduce the cerebral edema and improve the inflammation， and exert the neuroprotection effect.

To investigate the protective effect of Bupleurum chinense polysaccharide （BCP） on the aging model mice induced by D-galactose，and to elucidate its mechanism.

A total of 25 mice were randomly divide into control group（given saline）， model group （given 120 mg·kg^－1 D-galactose）， positive control group （given 120 mg·kg^－1 D-galactose and 100 mg·kg^－1 VE）， low dose of BCP group（given 120 mg·kg^－1 D-galactose and 200 mg·kg^－1 BCP）， high dose of BCP group （given 120 mg·kg^－1 D-galactose and 400 mg·kg^－1 BCP）. The mice in control group were injected intraperitoneally daily with saline， and the mice in the other groups were injected with D-galactose intraperitoneally to construct the aging models. The body weights， liver indexes，and kidney indexes of the mice in various groups were detected；the serum activities of superoxide dismutase （SOD） and glutathione peroxidase （GSH-Px） and the serum levels of malondialdehyde （MDA） of the mice in various groups were detected；HE staining was used to detect the morphology of liver tissue of the mice in various groups； Western blotting method was used to detect the expression levels of p53 and p16 proteins in kidney tissue of the mice in various groups.

After 7 weeks of continuous injection of D-galactose， compared with control group， the body weight of the mice in model group was significantly decreased （P<0.05）， and the liver index and kidney index were increased （P<0.05），the activities of SOD and GSH-Px in serum were decreased （P<0.01）， and the level of serum MDA was increased （P<0.01）； the expression levels of p53 and p16 proteins in kidney tissue were increased （P<0.01）； meanwhile， the hepatocyte edema was obvious， the liver tissue injury was severe. Compared with model group， the body weights of the mice in low and high doses of BCP groups were significantly increased （P<0.05），the liver indexes and kidney indexes were decreased （P<0.05）， the activities of SOD and GSH-Px in serum were increased （P<0.01）， the levels of serum MDA were decreased （P<0.01），and the expression levels of p53 and p16 proteins in kidney tissue were decreased （P<0.05 or P<0.01 ）； the morphology of liver cells was gradually normalized and the liver injury was alleviated.

BCP can inhibit the D-galactose-induced aging in the mice by inhibiting the oxidative stress and its downstream p53 and p16 signaling pathways.

To study the effects of osteoprotein on the osteoclast differentiation and p38-mitogen-activated protein kinase （p38-MAPK） signal pathway during the orthodontic tooth movement in the rats，and to explore the possible mechanism of osteoprotein in inhibiting the tooth movement during orthodontic treatment.

A total of 128 rats were randomly divided into control group and osteoprotectin group， and there were 64 rats in each group.Three days before modeling， the rats in osteoprotectin group were injected with recombinant human osteoprotectin （0.05 mg·kg^－1） submucosally on the left maxillary first molar，and the rats in control group were injected with the same amount of saline at the same site.The orthodontic models of rats were established by spring traction method.The moving distances of first molar of the rats at the 1st week，the 2nd week，the 3rd week， and the 4th week after stressing were measured.Tartrate-resistant acid phosphatase （TRAP） staining was used to determine the number of osteoclasts of the rats in two groups；immunohistochemical staining was used to determine the expression level of p-p38 on the pressure side of periodontal tissue of the rats in two groups；Western blotting method was used to determine the expression levels of receptor activators nuclear factor kappa B （RANK） and matrix metalloproteinase-9 （MMP-9） proteins in periodontal tissue of the rats in two groups.

There were no significant differences in the tooth movement distances， osteoclast number and p-p38 expression levels of the rats between two groups at the 1st week after stressing； at the 2nd week， the 3rd week， and the 4th week after stressing， the tooth movement distances， osteoclast number and p-p38 expression levels of the rats in osteoprotegein group were lower than those in control group （P<0.05 or P<0.01）； the expression levels of RANK and MMP-9 proteins in alveolar bone tissue of the rats in osteoprotegein group were lower than those in control group （P<0.01）.The number of osteoclasts（r=0.654）， the expression levels of RANK（r=0.576） and MMP-9（r=0.583） proteins in periodontal tissue of the rats were positively correlated with the expression level of p-p38 protein（P<0.05）.

Osteoprotectin can inhibit the generation and differentiation of osteoclasts during the orthodontic tooth movement in the rats， and inhibit the activation of p38-MAPK signal pathway， thereby inhibit the orthodontic tooth movement and enhance the anchoring effect.

A total of 80 rats were randomly divided into control group， model group， positive drug group and treatment group；there were 20 rats in each group.The rheumatoid arthritis model was established with type Ⅱ collagen（Col Ⅱ） induction.The rats in positive drug group were treated with diclofenac sodium sustained-release tablets，and the rats in treatment group were treated with triptolide.After treatment，the body weights， paw thickness and arthritis index scores of the rats in various groups were measured；HE staining was used to determine the pathomorphology of joint synovial tissue of the rats in various groups；immunohistochemical staining was used to determine the microvessel density（MVD） of vascular endothelial growth factor （VEGF） expression level in synovial tissue of the rats in various groups； ELISA method was used to determine the serum levels of VEGF of the rats in various groups；Western blotting method was used to determine the expression levels of PTEN， PI3K， AKT and phosphorylated-AKT （p-AKT） proteins in synovial tissue of the rats in various groups.

Compared with control group， the body weight of rats in model group was decreased （P<0.05）， the paw thickness was increased （P<0.05）， the MVD and expression level of VEGF in synovial tissue were increased （P<0.05）， the expression level of PTEN protein in synovial tissue was decreased （P<0.05）， and the expression levels of PI3K， AKT and p-AKT proteins were increased （P<0.05）.Compared with model group， the body weights of the rats in positive drug group and treatment group were increased （P<0.05），the paw thickness was decreased （P<0.05），the MVD and the expression levels of VEGF in synovial tissue were decreased （P<0.05）， the expression levels of PTEN protein were decreased （P<0.05）， and the expression levels of PI3K， AKT and p-AKT proteins were decreased （P<0.05）.Compared with positive drug group， the MVD and expression levels of VEGF in synovial tissue of the rats in treatment group were decreased （P<0.05）， the expression level of PTEN protein in synovial tissue was increased （P<0.05）， and the expression levels of PI3K， AKT and p-AKT proteins were decreased （P<0.05）.The paw thickness and arthritis index of rheumatoid arthritis rats were positively correlated with VEGF（r=0.564， r =0.492） and p-AKT（r=0.561， r=0.468） （P<0.05），and negatively correlated with PTEN （r=－0.437， r=－0.521）（P<0.05）；MVD was positively correlated with VEGF（r =0.587）， PI3K（r =0.567）， and p-AKT（r=0.601） （P<0.05）， and negatively correlated with PTEN （r=－0.502）（P<0.05）.

Triptolide can inhibit the angiogenesis and PTEN / PI3K / AKT pathway activation in the rheumatoid arthritis rats， thereby exerting its therapeutic effect on rheumatoid arthritis.

A total of 80 cases of thyroid follicular carcinoma tissue specimens and 80 cases of thyroid follicular adenoma tissue specimens confirmed by pathology were selected. Immunohistochemistry was used to detect the expressions of BTg-3 in thyroid follicular carcinoma and thyroid follicular adenoma tissues. Western blotting method was used to detect the expression levels of BTg-3 protein in thyroid follicular carcinoma CGTHW-1 cells and thyroid follicular adenoma Nthy-ori cells.The thyroid follicular carcinoma CGTHW-1 cells were randomly divided into blank control group （without transfection）， negative control group（transfected with negative control vector），and BTg-3 over-expression group （transfected with BTg-3 over-expression vector）.Plate clone experiment was used to detect the clone formation rates of the thyroid follicular carcinoma CGTHW-1 cells in various groups；Transwell chamber assay was used to determine the number of invasion and migration cells in various groups；Western blotting method was used to detect the expression levels of Ki67， proliferating cell nuclear antigen （PCNA）， Vimentin， epithelial cadherin （E-cadherin）， WNT1， β-catenin， glycogen synthesis kinase 3β （GSK3β） ，and phosphorylated GSK3β （p-GSK3β） proteins in the thyroid follicular carcinoma CGTHW-1 cells in various groups.

The positive expression rate of BTg-3 in thyroid follicular carcinoma tissue was lower than that in thyroid follicular adenoma tissue （χ² = 62.864， P<0.01）.The expression level of BTg-3 protein in the thyroid follicular carcinoma CGTHW-1 cells was lower than that in thyroid follicular epithelium Nthy-ori cells （t = 34.484， P<0.01）.Compared with blank control group and negative control group， the expression level of BTg-3 protein in the thyroid follicular carcinoma CGTHW-1 cells in BTg-3 over-expression group was increased （P<0.05）；the clone formation rate， number of invation cells and number of migration cells in the thyroid follicular carcinoma CGTHW-1 cells in BTg-3 over-expression group were decreased （P<0.05）；the expression levels of Ki67， PCNA， Vimentin， WNT1， β-catenin， and p-GSK3β proteins in the thyroid follicular carcinoma CGTHW-1 cells in BTg-3 over-expression group were decreased （P<0.05）；the expression level of E-cadherin protein was increased （P<0.05）.

The expression levels of BTg-3 in the thyroid follicular cancinoma tissue and cells are decreased. Over-expression of BTg-3 can inhibit the proliferation， invasion，and migration of thyroid follicular carcinoma cells， and inhibit the activation of WNT / β-catenin signaling pathway.

To observe the effect of total saponins of clematis （TSC） on the T lymphocyte subsets in peripheral blood and the serum levels of inflammatory factors in the rats with rheumatoid arthritis （RA）， and to illuminate its inhibitory effect on RA.

A total of 50 SD rats were randomly divided into control group （given normal saline）， model group （given normal saline）， low， medium， and high doses of TSC groups （given 50， 100， and 200 mg·kg^－1 TSC）， and there were 10 rats in each group. In addition to control group， the RA rat models were established in the other groups. The swelling degrees of ankle joint of the rats in various groups were measured and the arthritis indexes were calculated after 6 weeks of administration. Flow cytometry was used to detect the percentages of CD4⁺ T lymphocytes and CD8⁺ T lymphocytes in peripheral blood of the rats in various groups；ELISA method was used to detect the levels of serum tumor necrosis factor-α（TNF-α），interleukin-1β（IL-1β）， and interleukin-6（IL-6） of the rats in various groups； the spleen weights of the rats in various groups were measured and the spleen index was calculated.

Compared with control group， the arthritis index of the rats in model group was significantly increased （P<0.05）， the percentage of CD4⁺ T lymphocytes and the ratio of CD4⁺/CD8⁺ T lymphocytes were significantly increased （P<0.05）， the percentage of CD8⁺ T lymphocytes was significantly decreased （P<0.05）， and the levels of serum TNF-α，IL-1β，and IL-6 and the spleen index were significantly increased （P<0.05）. Compared with model group， the arthritis indexes of the rats in low， medium and high doses of TSC groups were significantly decreased （P<0.05）， the percentages of CD4⁺ T lymphocytes and the ratioes of CD4⁺/CD8⁺ T lymphocytes were significantly decreased （P<0.05）， the percentages of CD8⁺ T lymphocytes was significantly increased （P<0.05）， and the levels of serum TNF-α， IL-1β，and IL-6 and the spleen indexes were significantly decreased （P<0.05）. Compared with low dose of TSC group， the arthritis indexes of the rats in medium and high doses of TSC groups were significantly decreased （P<0.05）， the percentages of CD4⁺ T lymphocyte and the ratios of CD4⁺/CD8⁺ T lymphocytes were significantly decreased （P<0.05）， the percentages of CD8⁺ T lymphocytes were significantly increased （P<0.05）， and the levels of serum TNF-α， IL-1β and IL-6 and the spleen indexes were significantly decreased （P<0.05）. Compared with medium dose of TSC group， the arthritis index of the rats in high dose of TSC group were significantly decreased （P<0.05）， the percentage of CD4⁺ T lymphocytes and the ratio of CD4⁺/CD8⁺ T lymphocytes were significantly decreased （P<0.05）， the percentage of CD8⁺ T lymphocytes was significantly increased （P<0.05）， and the levels of serum TNF-α， IL-1β ，and IL-6 and the spleen index were significantly decreased （P<0.05）.

TSC can inhibit RA by regulating the percentages of CD4⁺ and CD8⁺ T lymphocytes and inhibiting the secretion of inflammatory cytokines in a dose-dependent manner.

To investigate the effect of long non-coding RNA （lncRNA）-colon cancer-associated transcript 2 （CCAT2） on the apoptosis of cervical cancer CaSki cells and its mechanism.

The human cervical cancer CaSki cells were divided into control group， over-expression CCAT2 group， sh-CCAT2 group， over-expression NC group and sh-NC group. The over-expression and interference vectors of CCAT2 were constructed and transfected into the cervical cancer CaSki cells in over-expression CCAT2 and sh-CCAT2 groups， respectively. The expression levels of CCAT2 mRNA in cervical cancer CaSki cells in various groups were detected by qRT-PCR method； MTT assay was used to detect the viability of cervical cancer CaSki cells in various groups；flow cytometry was used to detect the apoptotic rates of cervical cancer CaSki cells in various groups； qRT-PCR method was used to detect the expression levels of phosphatase and tensin homolog deleted on chromosome ten （PTEN） mRNA in cervical cancer CaSki cells in various groups；Western blotting method was used to detect the expression levels of PTEN， phosphoinositide 3-kinase （PI3K），phosphorylated PI3K（ p-PI3K）， protein kinase B （AKT），phosphorylated AKT （p-AKT）， B-cell lymphoma-2 （Bcl-2）， and Bcl-2 associated X protein （Bax） proteins in the cervical cancer CaSki cells in various groups.

Compared with control group， the cell proliferation rate and the expression levels of CCAT2 mRNA and Bcl-2， p-PI3K，and p-AKT proteins in the cervical cancer CaSki cells in over-expression CCAT2 group were significantly increased（P<0.05）， while the apoptotic rate and the expression levels of Bax protein， PTEN mRNA and protein were significantly decreased （P<0.05）. Compared with control group， the proliferation rate of cervical cancer CaSki cells and the expression levels of CCAT2 mRNA and Bcl-2， p-PI3K， and p-AKT proteins in the cervical cancer CaSki cells in sh-CCAT2 group were significantly decreased （P<0.05）； the apoptotic rate and the expression levels of Bax protein， PTEN mRNA and protein were significantly increased （P<0.05）. Compared with control group， the cell proliferation rate， apoptotic rate， and expression levels of CCAT2 mRNA and PTEN， Bcl-2， Bax， p-PI3K，and p-AKT proteins in the cervical cancer CaSki cells in over-expression NC group and sh-NC group had no significant differences（P>0.05）.

LncRNA-CCAT2 can inhibit the apoptosis of cervical cancer cells， and its mechanism may be related to the PTEN/PI3K/AKT pathway.

To explore the protective effect of ranolazine preconditioning on the H9C2 cardiomyocytes injured by hypoxia/reoxygenation， and to elucidate the mechanism of ranolazine in protecting myocardium of ischemia-reperfusion injury of the point of regulation autophagy view.

The H9C2 cells were randomly divided into normal control group， hypoxia/reoxygenation model group， ranolazine preconditioning group， ranolazine+Wortmannin group， and rapamycin group. The cells in normal control group didn’t receive any treatment， and the H9C2 cells in the other groups were pretreated with the corresponding drugs， and then were treated with hypoxia for 4 h and reoxygenation for 3 h. The viabilities of the H9C2 cells in various groups were detected by CCK-8 method； the levels of lactate dehydrogenase （LDH） in the H9C2 cells in various groups were detected by LDH kit； the positive expression rates of microtubule-associated protein light chain 3（LC3） in the H9C2 cells in various groups were analyzed by immunofluorescence method； Western blotting method was used to detect the expression level of LC3， mammalian target of rapamycin（ mTOR） and phosphorylated mTOR（ p-mTOR） in the H9C2 cells in various groups，　and the ratios of LC3Ⅱ/LC3Ⅰ and p- mTOR/ mTOR were calculated.

Compared with hypoxia/reoxygenation model group， the viabilities of H9C2 cells in ranolazine precoditioning group and rapamycin group were increased significantly （P<0.01）， the activities of LDH in the H9C2 cells were decreased significantly （ P<0.05）， and the positive expression rates of LC3 were increased significantly （ P<0.05）.The Western blotting results showed that compared with normal control group， the ratio of LC3 Ⅱ/LC3Ⅰ in the H9C2 cells in hypoxia/reoxygenation model group was increased significantly （P<0.01） and the ratio of p-mTOR/ mTOR was decreased significantly （P<0.01）.Compared with hypoxia/reoxygenation model group， the ratios of LC3 Ⅱ/LC3Ⅰ in the H9C2 cells in ranolazine preconditioning group and rapamycin group were increased significantly （P<0.01）， and the ratios of p-mTOR/mTOR were decreased significantly （P<0.01）.

Ranolazine preconditioning has a protective effect on the H9C2 cells injured by hypoxia/reoxygenation，and its mechanism may be related to its inhibition in mTOR protein phosphorylation and induction of the autophagy of H9C2 cells injured by hypoxia/ reoxygenation.

To detect the effects of nobiletin on proliferation， apoptosis， migration and invasion of the human prostate cancer DU145 cells， and to explore its mechanism.

The DU145 cells were cultured and divided into control and 0.8，1.6，3.2，6.4，12.8，25.6，and 51.2 μmol·L^－1 nobiletin groups. MTT method was used to detect the proliferation activities of human prostate cancer DU145 cells in various groups； cell scratch test was used to analyze the cell migration abilities of human prostate cancer DU145 cells in various groups；Transwell test was used to analyze the invasion abilities of human prostate cancer DU145 cells in various groups；flow cytometry was used to detect the apoptotic rates of human prostate cancer DU145 cells in various groups. The expression levels of phosphorylated c-Jun N-terminal kinase （p-JNK）， phosphorylated P38 mitogen activated protease （p-P38）， and phosphorylated extracellular regulated protein kinase （p-ERK） proteins in human prostate cancer DU145 cells in various groups were analyzed by Western blotting method.

Compared with control group， the proliferation activities of human prostate cancer DU145 cells in 25.6 μmol·L^－1 nobiletin group after treated for 24， 48， and 72 h were significantly decreased（P<0.01）； nobiletin inhibited the growth of DU145 cells in time- and concentration-dependent manners. Compared with control group， after treated for 48 h，the scratch healing rate and the number of invasion cells of human prostate cancer DU145 cells in 12.8 μmol·L^－1 nobiletin group were significantly decreased（P<0.01）， and the expression level of p-ERK protein was decreased （P<0.01）； the expression level of p-ERK protein in human prostate cancer DU145 cells in 25.6 μmol·L^－1 nobiletin group was decreased（P<0.01），the apoptotic rate was significantly increased（P<0.01）， and the expression levels of p-JNK and p-P38 proteins were increased（P<0.01）.

Nobiletin has the inhibitory effect on the growth of human prostate cancer DU145 cells and it can promote the apoptosis and inhibit the cell proliferation，migration and invasion. The changes of MAPK related protein expressions may be one of the mechanisms of the inhibitory effect of nobiletin on the growth of human prostate cancer DU145 cells.

To investigate the effect of neutrophil/lymphocyte ratio （NLR） in judgement of progrosis in the patients with non-small cell lung cancer（NSCLC） after treated with epidermal growth factor receptor tyrosine kinase inhibitors （EGFR-TKIs）， and to provide the evidence-based medical basis for the prognosis judgement of NSCLC patients.

PubMed， Cochrane Library， EMBASE， Web of Science and China National Knowledge Internet（CNKI） databases were searched for collecting the studies about the relationship between NLR and the prognosis of NSCLC patients treated with EGFR-TKIs. Meta-analysis was conducted on the literatures that met the inclusion criteria.

Eight studies contained 1 451 NSCLC patients were eventually included in the Meta-analysis. The progression-free survival（PFS）［hazard ratio（HR）=1.75， 95% confidence interval（95%CI）：1.24－2.48， Z=3.17， P=0.002］ and overall survival（OS）（HR=1.87， 95%CI：1.21－2.88， Z=2.84， P=0.005） of the NSCLC patients in high NLR group were significantly lower than those in low NLR group. The subgroup analysis results showed that after grouping by the ethnicity and NLR cutoff value， the PFS of NSCLC patients in the Asian（HR=1.70， 95%CI：1.14－2.53， Z=2.62， P=0.009）， the non-Asian（HR=2.00， 95%CI：1.27－3.15， Z=3.00， P=0.003） and the NLR cutoff value≥3.5（HR=2.21， 95%CI：1.54－3.17， Z=4.29， P<0.001） in high NLR group were significantly low than those in low NLR group.The sensitivity analysis results showed that 8 included studies were stable and reliable.

In the NSCLC patients after treated with EGFR-TKIs， the high level of NLR is associated with the poor prognosis of NSCLC patients.

To detect the expression levels of 17 kinds of gastric cancer-related cytokines in serum of the gastric cancer patients and study their correlations with miR-10b level， and to screen the tumor markers which were meaningful for the diagnosis of gastric cancer and to explore their application values in diagnosis of gastric cancer.

Based on the case-control study design， 34 cases of serum specimens from patients with definite histopathological diagnosis of gastric adenocarcinoma were selected as gastric cancer group， and 80 cases of serum specimens from healthy people who passed the physical examination were selected as control group. Luminex detection technology was used to detect the levels of gastric cancer-related cytokines in serum samples of the subjects in two groups. The tumor markers that were meaningful for the diagnosis of gastric cancer were screened from the following indexes such as macrophage inflammatory protein-3α （CCL20）， interleukin-2 （IL-2）， interleukin-4 （IL-4）， Interleukin-5 （IL-5）， interleukin-6 （IL-6）， interleukin-10 （IL-10）， interleukin-17A （IL-17A）， tumor necrosis factor-α （TNF-α）， interferon-γ （IFN-γ）， monocyte chemoattractant protein-4 （CCL13）， interferon-induced T cell alpha chemokine （CXCL11）， B-lymphocyte chemoattractant （CXCL13）， bone morphogenetic protein 4 （BMP4）， bone morphogenetic protein7 （BMP7）， hepatocyte growth factor （HGF）， granulocyte-macrophage colony stimulating factor （GM-CSF）， platelet derived growth factor-BB （PDGF-BB）.Quantitative real-time polymerase chain reaction（qRT-PCR） method was used to detect the serum miR-10b level of the patients in gastric cancer group；electrochemiluminescence method was used to detect the serum tumor markers carcinoembryonic antigen（CEA）， carbohydrate antigen 19-9（CA19-9） and carbohydrate antigen 72-4（CA72-4） levels of the patients in gastric cancer group.

Compared with control group， the serum levels of CCL20， IL-2， IL-4， IL-6， IL-17A， TNF-α， CCL13， BMP-4， and HGF of the patients in gastric cancer group were significantly increased （P<0.05）； the Logistic regression analysis results showed that the serum levels of CCL20， IL-2， IL-4， IL-6， IL-17A， TNF-α， CXCL11， CCL13， BMP-4， BMP-7， and HGF were associated with the risk of gastric cancer （ P<0.05）； the receptor operation characteristic curve（ROC） analysis results showed that the area under the curve （AUC） of CCL20 in diagnosis of gastric cancer was significantly better than combinnation diagnosis of the three tumor markers CEA， CA19-9，and CA72-4； When the cut-off value of CCL20 was 6.71 ng·L^－1， the diagnostic specificity， sensitivity， positive predictive value， negative predictive value， positive likelihood ratio， negative predictive ratio， positive likelihood ratio， negative predictive ratio and diagnostic accuracy were 76.5%， 91.3%， 78.8%， 90.1%， 8.74， 0.26，and 86.84%， respectively. The Spearman correlation analysis results showed that 17 kind of cytokines levels had poor correlations with miR-10b level， and the level of TNF-α had a certain correlation with miR-10b level（r=0.335， P=0.075）.

When 6.71 ng·L^－1 is used as the diagnostic cut-off value of the serum cytokine CCL20， its diagnostic sensitivity is significantly better than the tumor markers CEA， CA19-9 and CA72-4 commonly used in clinic， and the specificity is also higher.

To investigate the differentially expressed circRNAs in the colorectal cancer tissue and adjacent cancer tissue， and to elucidate the potential mechanism of inhibitory effect of hsa_circ_0043278 on colorectal cancer.

The data were downloaded from the GEO database and the GSE126094 chip data contained 10 samples of colorectal cancer tissues and 10 samples of adjacent cancer tissues. R software was used to select the differentially expressed circRNAs； the miRNAs that bound to differentially expressed circRNAs were found in the CSCD database；Perl software was used to predict the target genes of the miRNAs； GO biological function enrichment analysis and KEGG pathway enrichment analysis were performed in the target genes.

Compared with adjacent cancer tissue samples，there were 23 circRNAs with increasing expression levels （logFC>2 and P<0.05） and 36 circRNAs with decreasing expression levels （logFC<－2 and P<0.05） in the colorectal cancer tissue samples. The expression level of hsa_circ_0043278 was decreased significantly（logFC=－7.481 and P<0.05）. There were 66 miRNAs combined with hsa_circ_0043278，and the target genes were predicted. The GO enrichment analysis results showed that the target genes were mainly involved in Wnt-mediated cell signal transduction；the KEGG pathway enrichment analysis results showed that the target genes mainly enriched in PI3K-AKT signaling pathway.

The expression level of hsa_circ_0043278 in the colorectal cancer tissue samples is significantly decreased， which indirectly affect the functions of target genes such as UTP18， CLIP4 and STC2， and further affect the regulation of Wnt-mediated cell signal transduction and PI3K-AKT signaling pathway， and finally reduce the inhibitory effect on the colorectal cancer.

To summarize the data of a family with Angelman Syndrome（AS） and analyze their clinical and genetic characteristics， and to improve the clinicans’ understanding of AS.

The history， clinical manifestation results of auxiliary examination and genetic detection of two brothers and their relatives in the family with AS were collected， and the relevant literatures were reviewed.

The patient was a boy，aged 4 monthes and 3 days who presented developmental delay， dyskinesia， feeding difficulties； his second elder brother aged 5 years and 2 months who displayed language disorder， dyskinesia， mental retardation， inappropriate laughter， hyperactive， abnormal behavior， seizures and characteristic electroencephalogram（EEG）. The second generation gene sequencing results showed the patient and his second elder brother had a novel maternal nonsense mutation of the UBE3A gene（c.766 C>T）. The results of Sanger sequencing showed the mutation derived from their mother.

AS is a rare neurodevelopmental disorder. Its early clinical manifestations are atypical and need to be confirmed by molecular biological techniques.

To investigate the levels of serum transformed growth factor beta 1 （TGF-β1） in the bronchiolitis pediatric patients with different severities and its correlations with the indexes of tidal breathing pulmonary function，and to discuss the significance of TGF-β1_{in the diagnosis and treatment of bronchiolitis.}

A total of 84 cases of pediatric patients with bronchiolitis were selected as bronchiolitis group， and they were divided into mild bronchiolitis group （n=46） and moderate-severe bronchiolitis group （n=38） according to the severity of the disease.Another 20 healthy infants who underwent physical examination at the same time were selected as healthy control group.The levels of serum TGF-β1 and the ridal breathing pulmonary function indexes of the pediatric patients at acute and convalescent phases and the subjects in healthy control group were detected， and Pearson correlation analysis was used to analyze the correlations between the serum level of TGF-β1_{and the tidal breathing pulmonary function indexes.}

The serum levels of TGF-β1 of the pediatric patients in mild bronchitis group and moderate-severe bronchiolitis group at acute phase were significantly higher than those in healthy control group （P<0.01）. Compared with mild bronchitis group， the serum TGF-β1 level of the pediatric patients in moderate -severe bronchiolitis group at acute stage was significantly increased （P<0.01）. Compared with acute stage， the serum TGF-β1_{levels of the pediatric patients in mild and moderate-severe bronchiolitis groups at convalescence stage were significantly decreased （P<0.01）. The TPTEF/TE and VPEF/VE of the pediatric patients in mild and moderate- severe bronchiolitis groups were significantly lower than those in healthy control group （P<0.05）. Compared with mild bronchiolitis group， the TPTEF/TE and VPEF/VE of the pediatric patients in moderate-severe bronchiolitis group were significantly decreased （P<0.05）. There was no significant difference in the tidal volume per kilogram of body weight among the subjects in three groups （P>0.05）.The serum level of TGF-β1 of the bronchiolitis pediatric patients was negatively correlated with TPTEF/TE and VPEF/VE （r=－0.552， P<0.01； r=－0.517，P<0.01）.}

The serum level of TGF-β1 in the bronchiolitis pediatric patients can indirectly reflect the degree of airway obstruction， which can be used as a reference indicator for the condition judgment of bronchiolitis and the prognosis evaluation of lung function.

To study the changes of cardiac function indexes， changes of related factors levels in blood and complications such as cerebral infarction in the patients with valvular disease complicated with atrial fibrillation after valve replacement combined with radiofrequency ablation and left atrial appendage closure.

The clinical data of 128 patients were retrospectively analyzed and the patients were divided into two groups according to the patient’s operation types： mitral valve replacement combined with radiofrequency ablation and left atrial appendage closure treatment group （combined surgery group） and mitral valve replacement treatment group.The sinus rhythm restoration rates， cardiac function indexes， complications， levels of interleukin-1β （IL-1β）， interleukin-6（IL-6）， and tumor necrosis factor-α（TNF-α） and homocysteine in blood of the patients in two groups were detected， and the prognosis was evaluated.

Compared with mitral valve replacement treatment group， the sinus rhythm restoration rates of the patients in combined surgery group in the end of surgery， three months， six months and one year after surgery were increased （P<0.01）. There were no significant differences in the preoperative cardiac function indexes of the patients between mitral value replacement treatment group and combined surgery group （P>0.05）. One year after surgery，compared with mitral value replacement treatment group， the left atrial diameter （LAD） and left ventricular end-diastolic diameter （LVEDD） of the patients in combined surgery group were decreased（P<0.05）， the ejection fraction （EF） was increased（P<0.05），the incidence rates of complications were deceased （P<0.05）， and the levels of IL-1β， IL-6， TNF-α， and homocysteine in blood of the patients were significantly decreased（P<0.01）.

Valvular replacement combined with radiofrequency ablation and left atrial appendage closure in the treatment of patients with valvular disease complicated with atrial fibrillation has good effect，and it can improve the cardiac function of patients， decrease the levels of inflammatory cytokines and homocysteine， reduce the incidence of cerebral infarction and bleeding， and improve the life quality of patients.

To investigate the therapeutic effect of minocycline hydrochloride in the postmenopausal women with periodontitis and its influence in the levels of inflammatory and bone metabolism factors in the gingival crevicular fluid， and to clarify the possible mechanism of minocycline hydrochloride in the treatment of postmenopausal women with periodontitis.

According to the patients’ willing，eighty-six postmenopausal women with periodontitis were divided into minocycline hydrochloride group （observation group， n=43） and iodine glycerin group （control group， n=43）. The total effective rates of the patients in two groups were evaluated. The probing depth （PD）， plaque index （PLI）， and bleeding index （BI） of the patients in two groups were recorded and compared before treatment and 4 weeks after treatment. The levels of interleukin 6 （IL-6）， hypersensitive C-reactive protein （hs-CRP） and tumor necrosis factor-α （TNF-α） in gingival crevicular fluid of the patients in two groups before treatment and 4 weeks after treatment were measured by ELISA method and the levels of osteocalcin （OC） in gingival crevicular fluid of the patients in two groups were measured by electrochemiluminescence.

There were no significant differences in PD，PLI，BI and the levels of IL-6， hs-CRP， TNF-α and OC in gingival crevicular fluid of the patients in two groups before treatment （P>0.05）. Four weeks after treatment，PD，PLI，BI and the levels of IL-6， hs-CRP， TNF-α and OC in gingival crevicular fluid of the patients in two groups were significantly lower than those before treatment （P<0.05）； compared with control group， the total effective rate of the patients in observation group was significantly increased （P<0.05），PD， PLI and BI were significantly decreased（P<0.05），the levels of IL-6， hs-CRP，and TNF-α in gingival crevicular fluid were significantly decreased （P<0.05）， and the level of OC in gingival crevicular fluid decreased was significantly decreased（P<0.05）.

Minocycline hydrochloride has a significant effect on the treatment of postmenopausal women with periodontitis. It can improve the periodontal parameters.

To investigate the diagnostic value of apparent diffusion coefficient （ADC） and apparent diffusion coefficient ratio （rADC）of diffusion weighted imaging（DWI） in the differentiation of benign and malignant parotid tumors.

Eighty-four patients with parotid gland tumor confirmed by pathology were selected. According to the postoperative pathological types， the patients were divided into benign tumor group and malignant tumor group.The patients received preoperative routine MRI and DWI sequence scanning.HE staining was used to detect the pathomorphology of benign and malignant parotid gland tumors and their parotid gland tumor tissue with different pathological types，and the ADC of benign and malignat parotid gland tumors and their adjacent glands of and contralateral glands of were measured， respectively.The corresponding rADC were calculated.

There were 60 cases of benign parotid gland tumor and 24 cases of malignant parotid gland tumor；the benign tumor had the clear multi-boundary， and the mixed tumor and the adenomas were the most common types；the T2WI results showed the isostatic and low-signal lesions； the malignant parotid gland tumor was more common in mucoepidermoid carcinoma with irregular morphology， blurred boundaries and uneven signals， usually accompanied by invasion of peripheral structures and enlargement of cervical lymph nodes. Compared with benign parotid gland tumor，the ADC of parotid gland malignant tumor was decreased（P<0.05），and the rADC of contralateral glands and adjacent glands of parotid malignant tumor were decreased （P<0.05）；the ROC curve detection results showed that the area under the curves （AUC） of ADC， rADC of contralateral，glands， and rADC of adjacent glands were 0.859， 0.874，and 0.894， respectively.The AUC of ADC and contralateral glands and the AUC of ADC and adjacent glands were 0.874 and 0.894 calculated by combined multiple indexes.

The ADC and rADC of DWI can provide the reference for the diagnosis and differential diagnosis of benign and malignant parotid gland tumors， which has important clinical significance.

To establish a double antibody sandwich ELISA for rapid determination of varicella zoster virus（VZV） glycoprotein level， and to validate its specificity， linearity， accuracy， and precision.

Double antibody sandwich ELISA was established with the recombinant antibodies against VZV glycoprotein and used for the determination of VZV glycoprotein level. The antibody combination， antibody concentration， coating conditions， blocking conditions， enzyme-labled antibody reaction conditions，and chromogenic condition were optimizated in the experiment， and the established method was validated for the specificity， linearity， accuracy， and precision.

The optimal combination of the double antibody was the group of VZV-mAb-03 as the capture antibody at a concentration of 450.0 μg·L^－1 and VZV-mAb-HRP-02 as the enzyme-labeled antibody at a concentration of 0.1 μg·L^－1； the optimal buffer for antibody coating was 0.05 mol·L^－1 Tris-HCl（pH=8.7±0.1），while the optimal temperature and time for coating were 4 ℃ and overnight（18 h） respectively； the bovine serum albumin（BSA） at a concentration of 1% was served as the blocking agent， while the optimal temperature and time for blocking were 37 ℃ and 2 h， respectively； the optimal reaction time of VZV and enzyme-labeled second antibody with PBST were both 1 h at the temperature of 37 ℃， and the optimal temperature and time for color reaction were 37 ℃ and 15 min， respectively. It was proved that this method had no cross reactions with the other vaccines or ingredients. The quantitation scope was 500－2 600 PFU·mL^－1， regression coefficient（R² ）> 0.95； the in-batch recovery rate was 82.2%－115.7% ， and the in-batch coefficient of variable（CV）< 10.2%； the batch-to-batch recovery was 80.5%－118.6%， the CV in validation for inplate<17.4% and the CV inter-plate precisions<14.9%.

Double antibody sandwich ELISA suitable for determination of VZV glycoprotein level is established，which meets the requirements for quantitative determination， and might be used to determine the VZV antigen level during the development and production of VZV vaccine.