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Objective To explore the application of ZnxNi1-xS@DOX nanostructures in combined photothermal-chemotherapy treatment of tumors, and to provide new ideas for the study of tumor treatment methods. Methods The morphology and elementary composition of ZnxNi1-xS nanostructures were characterized by scanning emission electron microscopy and other methods. The photothermal properties of ZnxNi1-xS nanostructures were explored by infrared thermal camera. The release of DOX under different temperature and pH was investigated by fluorescence spectrophotometer. The biosafety of ZnxNi1-xS nanostructures and thein vitroantitumor properties of ZnxNi1-xS@DOX nanostructures were investigated by MTT assay. Results The as-prepared ZnxNi1-xS nanostructure had good dispersion and uniform size. The ZnxNi1-xS nanostructures had good photothermal effect. The drug release behavior showed temperature and pH responsiveness. The MTT assay showed that ZnxNi1-xS nanostructures were nontoxic to normal cells. ZnxNi1-xS@DOX nanostructures could significantly inhibit the growth of tumor cells under near-infrared illumination. Conclusion The as-prepared ZnxNi1-xS nanostructure has good biocompatibility, on the basis of which the ZnxNi1-xS@DOX nanostructures achieve good antitumor performance, which can be used for combined photothermal-chemotherapy treatment of tumors.

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Objective To investigate the effect of paeoniflorin(PF) on liver injury mediated by NLRP3 inflammasome pathway in db/db mice. Methods In this study, db/db mice were used as the mice model of type 2 diabetic while db/m mice were used as control group. The mice were randomly divided into six groups: db/m group,db/m+ PF group,db/db group,db/db + PF 25 mg/kg group,db/db + PF 50 mg/kg group,db/db + PF 100 mg/kg group. After 12 weeks of PF gavage,mouse serum samples were collected to detect the levels of alanine aminotransferase( ALT),aspartate aminotransferase( AST),total cholesterol( TC),triglyceride( TG) and free fat acid( FFA).HE staining,oil red O staining and Sirius red staining were used to observe the degree of pathological injury of liver. The expression of F4/80,α-SMA and Col Ⅲ protein was detected by immunohistochemistry. The expression of interleukin-1β( IL-1β), interleukin-18( IL-18), tumor necrosis factor-α( TNF-α), NOD-like receptor 3( NLRP3),apotpsis associated spck-like protein( ASC) and cysteinyl asparate specific proteinase-1( Caspase-1)was detected by Western blot. Results Compared with db/m group,the levels of serum ALT,AST,TC,TG and FFA increased in db/db group,and these indexes decreased after PF gavage. Compared with db/m group,histopathological examination of the liver revealed increased hepatic tissue lipid accumulation,inflammatory cell infiltration,and collagen deposition in db/db mice,and PF treatment reduced hepatic lipid accumulation,inflammatory cell infiltration,and collagen deposition. Meanwhile,compared with db/m group,the expression of proinflammatory cytokines( IL-1β,IL-18 and TNF-α),F4/80,α-SMA,Col Ⅲ,NLRP3,ASC and Caspase-1 protein in the liver of db/db mice increased,and the expression of these proteins decreased after gavage with PF. Conclusion PF inhibited the NLRP3 inflammatory pathway and attenuated liver inflammation and fibrosis in db/db mice.

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Objective The CRISPR/dCas9-SAM system was used to explore genes related to the proliferation of gastric cancer cells AGS, and their role in the occurrence and development of gastric cancer was analyzed. Methods sgRNA was designed for genes with differential expression between gastric cancer and normal gastric tissue, and a lentiviral library was obtained after packaging was constructed. The AGS cells at different time points after the library was infected with AGS cells were used as the screening pressure, and the AGS cells at three time points on days 0, 7 and 14 were collected. High-throughput sequencing analyzed sgRNA enrichment in AGS cells at different time points after infection to obtain differential genes related to AGS cell proliferation. Results Bioinformatics showed that compared with the 0 d group, 42 and 45 negative screening differential genes and 59 and 40 positive screening differential genes were obtained in the 7 d group and 14 d group, respectively. Among them, the 7 d group and the 14 d group had 11 genes in the negative screening and the positive screening. Conclusion In this study, 11 genes inhibiting the proliferation of AGS cells were screened, of which 5 were protein-coding genes and 6 were long non-coding RNA(lncRNA) genes. 11 candidate genes that promoted AGS cell proliferation were screened, of which 3 were protein-coding genes and 8 were lncRNA genes. It laid a foundation for further functional verification and comprehensive analysis of the occurrence and development process of gastric cancer.

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Objective To study the osteogenic effect of odontogenic biphasic calcium phosphate(BCP) in maxillary sinus in rabbits compared with Bio-Oss®and CERASORB®M β-tricalcium phosphate(β-TCP). Methods BCP was prepared by calcination of extracted teeth, the phase and element analysis were carried out, and its surface characteristics were observed. The osteogenic ability was evaluated by micro-CT and histological analysis in rabbit maxillary sinus model. Results The material was BCP composed of hydroxyapatite(HA) and β-TCP, the Ca/P ratio was 1.627, dentin tubular-like pores and dense granular structures were found on the surface of BCP. Animal experiments showed that the BCP group had more new bone formation and better quality of new bone. Conclusion The odontogenic BCP has good osteogenic effect in rabbit maxillary sinus.

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Objective To explore the effect of ATP binding cassette subfamily A member 1(ABCA1) on the invasion, migration and epithelial-mesenchymal transition(EMT) of human gastric cancer and investigate the underlying mechanism. Methods The expression of ABCA1 in human gastric cancer and its relationship with the prognosis of gastric cancer patients were analyzed by Bioinformatics analysisbased on network databases. RT-qPCR was performed to detect the expression of ABCA1 in 25 pairs of clinical gastric cancer tissue samples; RT-qPCR and Western blot were used to analyze the expression of ABCA1 in different human gastric cancer cell lines; Gene silencing technology was used to construct stable low-expressing ABCA1 gastric cancer cell line. Cell migration and invasion were determined by transwell assay; Western blot was used to detect the expression of related marker proteins in EMT and epidermal growth factor receptor(EGFR)/signal transducer and activator of transcription(STAT3) signaling pathway. Results The results of bioinformatics analysis indicated that ABCA1 was highly expressed in gastric cancer, which was positively correlated with advanced grade and stage as well as the poor prognosis of gastric cancer patients. RT-qPCR results showed that ABCA1 was highly expressed in gastric cancer tissues and gastric cancer cells HGC-27; The results of Transwell indicated that knockdown of ABCA1 could inhibit the invasion and migration of HGC-27 cells. Western blot results showed that the expression of mesenchymal cell marker proteins(Vimentin, N-Cadherin) and EMT-related transcription factors(Twist, Zeb1 and Snail) in the ABCA1 knockdown group was down-regulated, while the expression of epithelial cell marker protein E-Cadherin was up-regulated. Western blot results showed that compared with the control group, knockdown of ABCA1 inhibited the expression of EGFR downstream molecule STAT3, and reduced the phosphorylation level of EGFR and its downstream molecule STAT3. Conclusion ABCA1 is up-regulated in gastric cancer and is correlated with cell invasion, migration, EMT and indicate poor prognosis of gastric cancer patients. ABCA1 may affect the expression and activation of STAT3 by regulating EGFR, thereby inhibiting the invasion, migration and EMT of gastric cancer cells.

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Objective To study the correlation between eukaryotic translation initiation complex 4 f(EIF4 F) and clinical prognosis of patients with hepatocellular carcinoma(HCC). Methods By following up the clinical data of 743 HCC patients in the specimen bank, 94 HCC tissue specimens with complete follow-up information were selected, and the clinical data were collected. The paired tissue specimens were made into tissue chip for immunohistochemical staining. Image J was used to analyze the optical density value of tissue chip staining, and R4.0.5 software was used to conduct nonparametric test analysis, draw KM curve, Cox regression analysis, and Nomogram statistical analysis on experimental data and follow-up data. Results Phosphorylation of 4 EBP1 was significantly activated in HCC tissues(P<0.001), and the activation Phosphorylation of 4 EBP1 was associated with the clinical prognosis of HCC patients,P=0.038. Conclusion The activation of 4 EBP1 phosphorylation in tumor tissue predicts shorter overall survival time in HCC patients.

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Objective To investigate the effect of Qiyu Sanlong Decoction(QYSLD) on inhibiting lung cancer by regulating tumor immune microenvironment through PD-1 signaling pathway and on Th1 immune response in lung cancer bearing mice. Methods Lewis lung cancer cell line(LLC) was used to model subcutaneously implanted tumor in mice. 72 mice were randomly divided into Control group(n=18), cisplatin group(n=18), QYSLD group(n=18) and combined group(n=18).The general survival of mice was observedand recorded; the specific Th1 cell response to tumor antigen was detected by ELISA method; the proportion of PD-1 positive cells and the apoptosis of memory CD4+T cells in spleen T cells of mice were detected by flow cytometry; and the number and volume of pulmonary metastatic nodules were counted under an anatomical microscope after 21 days of continuous treatment. Results QYSLD could improve the general survival of mice bearing lung cancer, up-regulate the immune response of CD4+T cells and the specific killing effect of CD8+T cells. The proportion of apoptotic cells in PD-1 positive cells and CD4+memory T cells in spleen T cells of mice was down-regulated, and lung metastasis of mouse tumor cells was inhibited. Conclusion QYSLD can improve the survival status of mice bearing lung cancer and enhance Th1 immune response by mediating PD-1 signaling pathway, improve immune function and inhibit immune escape of tumor cells, thus inhibiting the metastasis of lung cancer.

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Objective To explore the effect of perampanel regulation of mitogen actived protein kinase(MAPK) signaling pathway after injection of nitroglycerin(NTG) induced migraine rats. Methods Thirty-two male SD rats were randomly divided into four groups with eight in each group: normal saline, NTG group, perampanel 50 μg/kg+NTG group, perampanel 100 μg/kg+NTG group. The time of scratching the head and the number of climbing the cage were observed, the periorbital pain threshold of the rat was measured by the fiber filament, and the photophobia of the rat was observed in the light and dark box experiment. Immunofluorescence was used to detect the rat TGs PACAP expression level; RT-qPCR was used to determine the expression level of PACAP mRNA and ELISA was used to detect the concentration of PACAP in rat blood. Meanwhile, Western blot was used to detect p-p38 MAPK,p-JNK,p-ERK protein expression level in rats TG. Results Compared with the control group, the periorbital withdrawal threshold decreased, the cage climbing behaviors and facial grooming behaviors increased, and the time spent in the light compartment in the light-aversive behaviors decreased in the NTG group, the expression levels of PACAP mRNA and protein increased(P<0.05). While, the perampanel 50 μg/kg+NTG group and the perampanel 100 μg/kg+NTG group increased the periorbital withdrawal threshold, reduced the cage climbing behaviors and facial grooming behaviors, increased the time spent in the light compartment and decreased the expressions of PACAP mRNA and protein in comparison with the NTG group(P<0.05). Compared with the control group, the expression levels of p-p38 MAPK, p-ERK and p-JNK in NTG group increased(P<0.05); compared with the NTG group, the expression levels of p-p38 MAPK, p-ERK and p-JNK decreased in perampanel 50 μg/kg+NTG group and the perampanel 100 μg/kg+NTG group(P<0.05). Conclusion Perampanel may relieve migraine symptoms by inhibiting MAPK signaling pathway.

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Objective To investigate the expression of E3 ubiquitin ligase F-box and WD-40 domain protein 7(FBXW7) in esophageal squamous cell carcinoma(ESCC) and its effect on its biological behavior. Methods The expression of E3 ubiquitin ligase FBXW7 in 171 cases of ESCC and 53 cases of paracancer tissues was detected by immunohistochemistry. The relationship between FBXW7 expression and survival prognosis and clinicopathological features of patients with ESCC was explored. The ESCC cells of the overexpressed FBXW7 group and the ESCC cells of the control group were constructed, and the effects of FBXW7 on the proliferation, migration and invasion of ESCC cells were detected by CCK-8 assay, plate clone formation assay, Transwell cell migration and invasion assay and cell scratches assay. Results The expression of FBXW7 in paracancer tissues was higher than that in ESCC tissues(P=0.002). ESCC with low FBXW7 expression had lower differentiation degree(P=0.011) and TNM stage was earlier in ESCC patients with high FBXW7 expression(P=0.032), and the survival time of patients with high FBXW7 expression was longer than that of patients with low FBXW7 expression(P=0.030). CCK-8 assay results showed that the absorbance of cells in overexpressed FBXW7 group was lower than that in control group(P<0.001), and plate clone formation assay showed that the colony number of overexpressed FBXW7 group was lower than that of the control group(P<0.001). Cell scratches assay results showed that the cell migration rate of the overexpressed FBXW7 group was lower than that of the control group(P<0.001), Transwell cell migration and invasion assay showed that the number of cell migration and invasion in the overexpressed FBXW7 group was lower than that in the control group(P<0.001). Conclusion The expression of FBXW7 is low in ESCC and low expression of FBXW7 suggests poor prognosis. Overexpression of FBXW7 can inhibit the proliferation, migration and invasion of ESCC cells.

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Objective To observe the effect of sodium fluoride(NaF) and 2-(2-benzofuranyl) 2-imidazoline(2-BFI) on mitochondrial morphology, function, and apoptosis of SH-SY5 Y cells, and to explore the protective effect of 2-BFI on NaF-induced injury. Methods Different concentrations of NaF(0.5, 1.0, 1.5, 2.0, 4.0 mmol/L) were used to treat SH-SY5Y cells for 24 h,and then Cell Counting Kit-8( CCK-8) assay was used to screen the proliferation ability of SH-SY5Y cells; SH-SY5Y cells were treated with Na F and different concentrations of 2-BFI( 2. 5,5. 0,10. 0,20. 0,40. 0 μmol/L) for 24 h,and the proliferation ability of SH-SY5Y cells was detected by CCK-8 assay. The transmission electron microscope was used to observe the mitochondrial morphology,the mitochondrial function was detected by ATP detection kit and Mito-Tracker Red CMXRos kit,cell apoptosis was detected by TUNEL staining,and the expressions of Bcl-2,Bax,and cleaved caspase-3 protein were detected by Western blot. Results The cell survival rate of SH-SY5Y gradually decreased with the increase of Na F concentration.The selected Na F was 2. 0 mmol/L. 2-BFI treatment for 24 h could improve the cell survival rate. At 10 mmol/L,the cell survival rate was higher( 99. 169%),and 10 mmol/L was selected as the 2-BFI follow-up experimental concentration. Compared with the control group,the mitochondrial morphology of the Na F group changed,ATP levels and mitochondrial membrane potential( Δψm) decreased,the apoptosis rate increased,Bcl-2 protein content decreased,expression of Bax and cleaved caspase-3 protein increased. Compared with Group Na F,2-BFI administration improved mitochondria morphology,increased ATP levels and Δψm,reduced cell apoptosis rate,increased Bcl-2 protein content,and reduced Bax,cleaved caspase-3 protein expression. Conclusion 2-BFI can improve mitochondrial morphology and function,reduce cell apoptosis,and play neuroprotective role in fluorosis SH-SY5Y cells.

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Objective To study the effect of EFHD2 protein deletion in Sertoli cells on Occludin, a component of tight junction protein and the localization and expression of EF-hand domain family member D2(EFHD2) in mouse testis. Methods Total RNA and protein were extracted from adult miceʼs heart, liver, spleen, lung, kidney, brain and testis tissues. The mRNA and protein levels of EFHD2 in each organ tissue were detected by qRT-PCR and Western blot. Immunofluorescence and immunohistochemistry detected the localization and expression of EFHD2 in testicular tissues. SiRNA interference was used to reduceEFHD2in Sertoli cells to detect Occludin protein expression. Results qRT-PCR showed that the expression ofEFHD2was the highest in the testis. Western blot results showed that the expression level increased in testis tissue. Indirect immunofluorescence and immunohistochemistry results showed that the protein was mainly distributed in Sertoli cells and co-localized with cytoskeletal Vimentin, indicating that the protein was expressed in Sertoli cells.After the decrease of EFHD2 protein expression, Occludin protein expression also decreased. Conclusion The expression ofEFHD2protein in the testis is relatively high, mainly distributed in Sertoli cells of the testis, co-localized with Vimentin, and can affect the normal expression of tight junction protein Occludin. It is suggested that EFHD2 can promote and maintain the junction structure of Sertoli cells and provide a stable microenvironment for spermatogenesis.

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Objective To investigate the effect of teriparatide(TPTD) on the generation of MC3 T3-E1 cells towards osteogenic differentiationviathe Wnt3 a/β-catenin pathway in a high-glucose environment. Methods The experiment was divided into five groups: low glucose group, low glucose+TPTD group, high glucose group, high glucose+TPTD group, high glucose+TPTD+Wnt3 a inhibitor G244-LM group. Cell proliferation activity was detected by Calcein-AM and CCK-8 assay, cell mineralized nodule formation was observed by ALP and alizarin red staining, and actin formation was analyzed by immunofluorescence assay. Real-time PCR was performed to detectWnt3 a,β-catenin,Tcf1,OPG and COLⅠ mRNA expression. Results TPTD had no significant effect on the proliferative activity of MC3 T3-E1 cells under high glucose condition.The ALP staining area, protein activity and alizarin red staining area of the cells in the low glucose+TPTD group were higher than those in the other four groups(P<0.05); the high glucose group was lower than the low glucose group(P<0.05); the high glucose+TPTD group was higher than the high glucose group and the high glucose+TPTD+G244-LM group(P<0.05). The cytoskeleton in the low glucose+TPTD group was the clearest; the cytoskeleton was less clear in both the high glucose and high glucose+TPTD+G244-LM groups than in the high glucose+TPTD group. Genes such asWnt3 a,β-catenin,Tcf1,OPG and COLⅠ had the highest mRNA levels in the cells of the low glucose+TPTD group(P<0.05); the mRNA levels of all genes were higher in the low glucose group than thosein the high glucose group(P<0.05); the mRNA levels of all genes in the cells of the high glucose+TPTD group were higher than those in the high glucose group and the high glucose+TPTD+G244-LM group(P<0.05). Conclusion High glucose inhibited osteoblast differentiation, and TPTD promoted osteoblast differentiation in high glucose environment by regulating Wnt3 a/β-catenin pathway.

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Objective To investigate the effect of overexpression of tyrosine kinase receptor 1(Tie-1) in cervical cancer cells on the malignant biological behavior of tumor-related endothelial cells(TRECs). Methods Immunohistochemical method was used to detect the expression of Tie-1 in cervical cancer cells(CCCs) and TRECs of 96 patients with cervical cancer, and to analyze the correlation between the expression of Tie-1 in TRECs and clinicopathological features and prognosis of patients. HeLa cells overexpressing Tie-1(Hela-Tie1 OE) were constructed, and HeLa-Tie1 OE was co-cultured with human umbilical vein endothelial cells(HUVECs) by Transwell cell co-culture method to obtain cervical cancer TRECs. Western blot was used to analyze Tie-1 protein expression. The migration, invasion and tubulogenesis of TRECs were detected by cell scratch assay, Transwell invasion and migration assay and tubulogenesis assay. Results The expression of Tie-1 was positively correlated with CCCs and TRECs in 96 cervical cancer patients. The positive expression rate of Tie-1 in TRECs of patients with stage(FIGO) ⅠB2-ⅡA, tumor diameter ≥4 cm, cervical muscle invasion depth ≥1/2 full layer, adenocarcinoma, medium and low differentiation cervical cancer was higher than that of patients with ⅠA 1-ⅠB 1, tumor diameter<4 cm, cervical muscle invasion depth<1/2 full layer, squamous carcinoma, and high differentiation cervical cancer, respectively. The differences were statistically significant(P<0.05). The expression of Tie-1 in TRECs of cervical cancer patients had no significant correlation with age, lymph node metastasis and lymphatic space invasion. The positive expression of Tie-1 in cervical cancer TRECs was negatively correlated with 5-year progression-free survival time and overall survival time(P<0.05). The Tie-1 expression of TRECs obtained by co-culture of Hela-Tie1 OE and HUVECs was up-regulated, and the invasion, migration and tubulogenesis of TRECs cells were enhanced. Conclusion The high expression of Tie-1 in TRECs of cervical cancer is related to FIGO stage, tumor diameter, degree of differentiation, depth of cervical muscular invasion, type of cervical cancer, and poor prognosis of patients. Overexpression of Tie-1 can promote TRECs invasion, migration and tubulogenesis.

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Objective To study the new target and molecular mechanism of E3 ubiquitin ligase Ring finger 43(RNF43) in human gastric adenocarcinoma cell AGS. Methods The transfection efficiency of RNF43 in AGS cell was observed by fluorescence microscopy; the potential interaction between RNF43 and endogenic phosphates and tensin homologue deleted on chromosome ten(PTEN) in cancer cells was analyzed by Western blot; the specific interaction between RNF43 and PTEN and the ubiquitination of PTEN by RNF43 was verified by CO-IP; the apoptosis, proliferation and migration ability of gastric cancer cells was determined by flow cytometry, CCK-8 assay and scratch test. Results Cytofluorescence results showed that exogenous RNF43 was successfully overexpressed in AGS cells; RNF43 interacts with PTEN in AGS cells, and RNF43 could ubiquitinate PTEN through K63 ubiquitination and increase the protein of PTEN expression level. The results of cell function experiments showed that RNF43 could accelerate the apoptosis of AGS and inhibit the proliferation and migration of AGS by targeting and regulating the functional level of PTEN. Conclusion RNF43 inhibits the proliferation of AGS cellviaits functionality of ubiquitination towards PTEN.

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Objective To explore the mechanism of nuclear factor E2-related factor 2(Nrf2) in bleomycin(BLM)-induced DNA damage of lung cancer cells. Methods The fluorescence intensity of tissue reactive oxygen species(ROS) in human lung cancer cells A549 and H1299 was detected by a reactive oxygen species fluorescent probe(DCFH-DA). DNA damage of A549 and H1299 cells treated with 5 μg/ml BLM was detected by Western blot and micronucleus. Then, the Nrf2-low expression transient siRNA and the stably shRNA-transformed lung cancer cells were respectively constructed to detect the DNA damage after BLM. The interaction between Nrf2 and Rad51 in cells was detected by co-immunoprecipitation method. Results Under the action of BLM, ROS levels in A549 and H1299 cells increased significantly, and Nrf2 entered the cell nucleus for activation(P<0.01). Under the effect of BLM, resveratrol(Res) stimulated A549 and H1299 cells to up-regulate the protein expression of Nrf2(P<0.01), while the expression of DNA damage marker protein γ-H2 AX decreased(P<0.01). On the contrary, after interfering the protein expression of Nrf2 in A549 and H1299 cells with siRNA, the protein expression of γ-H2 AX in A549 and H1299 cells was significantly up-regulated under the action of BLM(P<0.01). Western blot results showed that the expression of homologous recombination repair protein Rad51 and the expression of Nrf2 protein in A549 cells with low Nrf2 expression were significantly down-regulated(P<0.01). Conclusion The antioxidant transcription factor Nrf2 in tumor cells can interact with Rad51 to regulate the expression of Rad51 and reduce BLM-induced cellular DNA damage.

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Objective To investigate the co-labeling of c-Fos protein expression and oxytocin(OXT) neurons in related brain regions in mice injected with dexmedetomidine(Dex) by intraperitoneal injection. Methods 8-10 week old C57 mice were divided into Saline group and Dex group. The Saline group was intraperitoneally injected with normal saline, the Dex group was intraperitoneally injected with Dex(100 μg/kg), the control group was given normal saline(Saline group), and the experimental group was given Dex 100 μg/kg(Dex group). 120 minutes after intraperitoneal injection, the brains of two groups of mice were collected by perfusion, and thenfrozen section and immunofluorescence microscopy imagingwere performed. Results Immunofluorescence c-Fos protein expression in Dex group and Saline group. The results showed that compared with the Saline group, the Dex group had higher thalamotomy in the supraoptic nucleus(SON), ventrolateral preoptic nucleus(VLPO), subfornical organ(SFO), solitary tract(SOL) and area postrema(AP), and the difference was statistically significant.C-Fos in the locus coeruleus(LC), lateral hypothalamic area(LH), laterodorsal tegmental nucleus(LDT), parabigeminal nucleus(PB), ventral tuberomammillary nucleus(TMN) and motor cortex M1/M2 protein expression decreased, and the difference was statistically significant(P<0.05). There was no difference in c-Fos protein expression in parathalamic nucleus(PVN) and suprachiasmatic nucleus(SCN). The results of the co-labeling group of OXT neurons and c-Fos protein in the PVN and SON brain regions showed that: compared with the Saline group, there was no significant difference in the co-labeling of OXT neurons and c-Fos protein in the PVN and SON brain regions of the Dex group. Conclusion The immunofluorescence co-labeling experiment shows that Dex can activate sleep-promoting and other related brain regions, and has a certain inhibitory effect on wake-promoting and exercise-related brain regions, however, it does not affect the co-labeling of c-Fos protein and OXT neurons in PVN and SON brain regions.

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Objective To investigate the effect of Rotundic acid(RA) on proliferation, migration and invasion ability of human lung adenocarcinoma cells as well as its possible mechanisms. Methods Human lung adenocarcinoma A549 and PC9 cells were divided into control group, blank control group, solvent group and 20, 40, 60, 80 μmol/L RA groups. CCK-8 assay and scratch assay were used to detect the proliferation and horizontal migration of human lung adenocarcinoma cells. Transwell migration assay and Transwell invasion assay were used to detect the longitudinal migration and invasion ability of A549 and PC9 cells in each group. The protein expression levels of janus kinase 2(JAK2) and signal transducer and activator of transcription 3(STAT3) in the supernatants of A549 and PC9 cells were detected by ELISA. The mRNA expression levels of JAK2 and STAT3 were detected by RT-PCR. Statistical analysis was made on the differences among groups in each index. Results After RA treatment on human lung adenocarcinoma cells, compared with the control group, the proliferation activity of A549 and PC9 cells in the experimental groups decreased(P<0.05), the number of cells crossing polycarbonate membrane and matrix glue decreased(P<0.05), the expression of JAK2 and STAT3 proteins in cell supernatant decreased(P<0.05), and the mRNA expression of JAK2 and STAT3 decreased(P<0.05). The decrease of the above indices was concentration-dependent and had statistical significance(P<0.05).Compared with the control group, the proliferation activity of A549 and PC9 cells in the solvent group showed no significant difference. Conclusion RA may inhibit the proliferation, migration and invasion of human lung adenocarcinoma A549 and PC9 cellsin vitro, possibly through the inhibition of JAK2/STAT3 pathway.

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Objective To investigate the expression of alpha-actinin-4(ACTN4) and the effect on cell proliferation in esophageal squamous cell carcinoma(ESCC). Methods The expression of ACTN4 in ESCC tissues and paired normal tissues was detected by immunohistochemistry, and the correlation between ACTN4 and the clinicopathologi-cal features was analyzed statistically. The ACTN4 shRNA lentiviral plasmids were constructed, and the stable ECA109 strain with ACTN4 shRNA knockdown was established using lentivirus packaging technology. The knockdown efficiency on protein level was checked by Western blot,and cell proliferation was detected by colony formation assays. The downstream target proteins were validated in ESCC cell line ECA109 based on the previous proteomics analyses in melanoma cell line A375 with or without ACTN4 shRNA knockdown. Results The expression of ACTN4 in ESCC tissues was significantly higher than that of normal tissues. ACTN4 shRNA stable knockdown ECA109 cell strains were successfully constructed. The results of colony formation assays showed that ACTN4knockdown inhibited the cell proliferation and down-regulated NADH ∶ Ubiquinone oxidoreductase core subunit V1( NDUFV1) protein expression in ECA109 cells. Conclusion Upregulation of ACTN4 in ESCC cells promotes the cell proliferation and enhances the protein expression of NDUFV1.

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Objective To reveal the effect of lipopolysaccharide-binding protein(lbp) gene on sepsis in rats and its important regulatory mechanism. Methods The acute liver inflammatory injury model was induced by lipopolysaccharide(LPS). The levels of serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were detected by biochemical analyzer to evaluate the liver injury, and the proinflammatory factors TNF-α and IL-6 were detected by RT-PCR to evaluate the level of inflammation, liver hematoxylin-eosin staining was used to observe the infiltration of inflammatory cells and hepatocyte injury in liver tissue. RNA-Seq was performed to compare the transcriptome expression differences betweenlbpgene knockout(lbp-/-) rats and wild-type(WT) rats to excavate the downstream genes regulated bylbpin LPS induced liver inflammatory injury. The biological process and signal pathway were explored by GO function annotation and KEGG enrichment analysis. RT-PCR was used to verify the mRNA expression level of differential genes. Results Compared with WT group, the levels of ALT, AST and pro-inflammatory factors in serum oflbp-/-rats decreased(P<0.05), and histological observation showed that the infiltration of inflammatory cells decreased. Transcriptomic analysis of liver tissue showed that 168 genes were differentially expressed afterlbp-/-(P<0.05), among whichCyp7 a1、Cyp4 a2and other up-regulated genes were enriched in peroxisome proliferator activated receptor(PPAR) signal pathway and steroid hormone synthesis pathway(P<0.05). Conclusion lbp-/-may participate in biological processes such as bacterial clearance and lipid metabolism by promoting PPAR signal pathway, so as to reduce liver inflammatory injury and slow down the occurrence and development of sepsis.

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Objective To explore the effects of Blonanserin on hippocampal neurons damage and PI3 K/AKT/GSK3β signaling pathways in rats with MK-801 induced schizophrenia. Methods The schizophrenia models were induced by one-time intraperitoneal injection of MK-801. A total of 48 SD rats were randomly divided into normal group, model group, Blonanserin group(1 mg/kg Blonanserin) and risperidone group(0.54 mg/kg risperidone), with 12 cases in each group. The behaviors of rats were observed by stereotyped behavior assay, open field test and Morris water maze. The pathological damage of hippocampus was observed by HE staining, which was scored. The levels of serum glucose-lipid metabolism indexes were detected. The mRNA and proteins of phosphatidylinositol 3-kinase(PI3 K), protein kinase B(AKT) and glycogen synthase kinase 3β(GSK3β) in hippocampal tissues were detected by real-time fluorescent quantitative PCR and Western blot. Results Compared with model group, scores of stereotyped behaviors, total distance in open field test, escape latency and score of pathological damage in hippocampus decreased in Blonanserin group and risperidone group, while times of crossing platform original site, levels of PI3 K, AKT and GSK3β mRNA, and phosphorylation levels of PI3 K, AKT and GSK3 increased(P<0.05). However, there was no significant difference in the above indexes between Blonanserin group and risperidone group. The differences in levels of fasting blood glucose(FPG), glycosylated hemoglobin(HbA1 c), triglyceride(TG), total cholesterol(TC), high-density lipoprotein(HDL) and low-density lipoprotein(LDL) among normal group, model group and Blonanserin group was not statistically significant. However, levels of FPG, HbA1 c, TG and TC in risperidone group were higher than those in the other three groups(P<0.05), and there was no significant change in HDL or LDL. Conclusion Blonanserin may protect schizophrenia rats from hippocampal neurons damage, improve cognitive function, learning and memory ability by activating PI3 K/AKT/GSK3β signaling pathways.

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Objective To investigate the correlation between nocturnal sleep duration combined with snoring in the first trimester of pregnancyand small for gestational age(SGA), large for gestational age(LGA). Methods Multivariate Logistic regression model was used to analyze the association between nocturnal sleep duration, snoring, their combined effects and SGA, LGA. Results Compared to nocturnal sleep duration 7 to 9 h in the first trimester of pregnancy, sleep duration<7 h was positively correlated with SGA in male newborn(OR=4.22, 95%CI: 1.69-10.52); After stratified by snoring, the sleep duration of snoring women<7 h was positively correlated with SGA(OR=5.68,95%CI: 1.02-31.51), and the sleep duration of non-snoring women<7 h was positively correlated with LGA(OR=2.10, 95%CI: 1.16-3.81). Conclusion Sleep duration<7 h in the first trimester of pregnancy is a risk factor for SGA and LGA, and snoring may enhance the association between sleep duration<7 h in the first trimester of pregnancy and SGA. Pregnant women should keep adequate nocturnal sleep duration to reduce the risk of abnormal neonatal weight.

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Objective To investigate the association between adiposity rebound and metabolic abnormalities in preschools. Methods A prospective cohort study was designed on the basis of the Maanshan birth cohort. Venous blood samples were collected at 5 to 6 years of age to detect metabolic indicators. 2022 children aged 0 to 6 years with ≥8 consecutive measurements were enrolled. χ2test and Logistic regression model were used to analyze the data. Results The detection rate of abnormal metabolism in preschool children was 16.9%, and the risk of metabolic abnormalities in preschool children with high BMI level at the AR time point and earlier AR time phase was 2.59 and 1.82 times that of the normal group respectively. Conclusion High AR level and earlier AR phase can increase the prevalence of metabolic abnormalities in preschool children.

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Objective To explore the relationship between geriatric nutritional risk index(GNRI) and perioperative nutritional status, postoperative recovery and complications in elderly patients with gastric cancer. Methods In this retrospective study, 212 elderly patients(aged≥60 years) with gastric cancer who underwent gastrectomy were recruited. GNRI was used to retrospectively assess the patients′ preoperative nutritional status, and analyze the relationship between GNRI and perioperative nutritional status, postoperative recovery and complications.The ROC curve was applied to explore the value of GNRI in predicting postoperative complications. Results The incidence of preoperative nutritional risk in elderly patients undergoing gastric cancer surgery was 45.07%. Compared with the patients whose GNRI>98 points, the patients whose GNRI≤98 points had different degrees of decrease in serum total protein, albumin, prealbumin, hemoglobin and lymphocyte counts before surgery, day 1 and day 5-8 after surgery(P<0.05).The patients whose GNRI<92 points had longer postoperative hospital stay than those with GNRI>98 points(P<0.05).With the decrease of GNRI scores, the incidence of complications showed an upward trend(P<0.001). The multivariate analysis of the relationship between GNRI and postoperative complications showed that TNM staging of III-IV and GNRI<92 points were independent risk factors for complications. GNRI had a good predictive value for the occurrence of complications(AUC=0.639, 95%CI: 0.570-0.703,P=0.001, Cut-off value: 92.21). Conclusion GNRI can be used for preoperative nutritional assessment for elderly gastric cancer patients. Patients with GNRI<92.21 points should be actively given nutritional therapy to improve perioperative nutritional status, speed up postoperative recovery, and reduce the occurrence of complications.

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Objective To investigate the expression and prognosis of urokinase plasminogen activator(PLAU) and v-akt murine thymoma viral oncogene homolog 1(AKT1) in oral squamous cell carcinoma(OSCC) and normal tissues and the correlation of PLAU and AKT1 in OSCC tissues. Methods The expression levels of PLAU and AKT1 in 70 cases of oral squamous cell carcinoma and 50 cases of normal tissues were detected by immunohistochemical method, and the correlation between PLAU and AKT1 expression and clinicopathological features and prognosis as well as the correlation between PLAU and AKT1 expression in OSCC tissues was analyzed, and the results were further verified by bioinformatics database. Results The expression of PLAU and AKT1 in OSCC tissues was higher than that in normal tissues(P<0.05),Kaplan-Meier analysis showed that patients with low PLAU and AKT1 expression had longer survival time than those with high AKT1 expression(P<0.05),Spearman rank sum correlation test showed that there was a strong correlation between PLAU and AKT1 expression in OSCC tissues(r=0.357,P<0.05),GEPIA bioinformatics database analysis results are consistent with experimental results. Conclusion PLAU and AKT1 are highly expressed in OSCC tissues and are associated with poor prognosis of patients. There is a correlation between PLAU and AKT1 in OSCC tissues.

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Objective To observe the dynamic changes of reactive oxygen species(ROS), isocitrate dehydrogenase 2(IDH2) and sirtuin 3(SIRT3) in patients with acute myeloid leukemia(AML) before and after chemotherapy, and to explore the differences of ROS, IDH2 and SIRT3 levels in different AML patients and their application value in clinical diagnosis and efficacy evaluation. Methods Bone marrow and peripheral blood samples were taken from 20 newly diagnosed AML(non-APL) patients who had not received chemotherapy(new diagnosis group); bone marrow and peripheral blood samples were taken from 40 AML(non-APL) patients who had received chemotherapy(by Treatment group); 20 normal human bone marrow and peripheral blood samples were taken as control(control group). The expression of ROS in bone marrow mononuclear cells and mitochondria of bone marrow mononuclear cells in each group was observed under a fluorescence microscope, and the expressions of IDH2 and SIRT3 in peripheral blood of each group were determined by ELISA. The expression of AML1-ETO fusion gene in patients was summarized, and the expression of SIRT3 in different groups was compared. The expression of SIRT3 in different treatment effects and treatment courses in the treated patients were compared. Results The expression of mitochondrial ROS in bone marrow mononuclear cells and cells in the three groups was higher in the newly diagnosed group than that in the control group(P<0.05), and lower in the treated group than that in the newly diagnosed group(P<0.05). The expression of IDH2 in peripheral blood of the three groups was higher in the newly diagnosed group than that in the control group(P<0.000 1), and lower in the treated group than that in the newly diagnosed group(P<0.05). After peripheral blood samples were diluted 5 times, the expression of SIRT3 in the newly diagnosed group was lower than that in the control group(P<0.000 1); the expression of SIRT3 in the treated group was higher than that in the newly diagnosed group(P<0.01). The SIRT3 expression of fusion gene-positive patients was higher than that of fusion gene-negative patients; the SIRT3 expression of partial remission patients was higher than that of complete remission patients; the SIRT3 expression of patients with complete remission in the subgroup of previously treated patients was higher than that of patients with a longer treatment course than those with a short treatment course. Conclusion SIRT3 is an important protein that regulates metabolic function in the mitochondria of cells. Its expression is down-regulated in patients with acute myeloid leukemia, and the expressions of IDH2 and ROS increased accordingly. In AML, the abnormal expression of SIRT3 can accelerate the occurrence and development of tumor and reduce the therapeutic effect.

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Objective To explore the occurrence and related factors of liver fibrosis in patients with End-stage renal disease(ESRD). Methods A total of 83 ESRD patients were included in the study. Transient elastography was used to diagnose whether hepatic fibrosis occurred or not. According to the occurrence of hepatic fibrosis or not, the included patients were divided into a non-hepatic fibrosis group(n=37) and a hepatic fibrosis group(n=46). The demographic data and clinical laboratory indexes of the two groups were compared. Statistically significant variables were selected and included in the multivariate Logistics stepwise regression analysis to explore the influencing factors of liver fibrosis in ESRD patients. Results The prevalence of liver fibrosis in ESRD patients was 55.42%. Compared with the non-hepatic fibrosis group, the hepatic fibrosis group had lower white blood cells, erythrocyte sedimentation rate, complement C3 and C4(P<0.05). Multivariate Logistics analysis showed that complement C4 level(OR=0.930, 95%CI: 0.872-0.992,P=0.028) and higher lactate dehydrogenase(LDH) level(OR=1.016, 95%CI: 1.005-1.027,P=0.004) were the independent influencing factors of liver fibrosis in ESRD patients. Conclusion The probability of liver fibrosis in ESRD patients is high. Serum complement C4 and LDH are independent influencing factors of liver fibrosis. Dynamic monitoring of serum complement C4 and LDH levels is conducive to target liver fibrosis in ESRD patients.

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Objective To investigate the value of cranial ultrasonography in the diagnosis of periventricular-intraventricular hemorrhage(PIVH) and analyze the risk factors of PIVH in neonates. Methods 163 neonates hospitalized in the neonatal intensive care unit were examined by bedside cranial ultrasonography(CUS), The ultrasonography of PIVH were reviewed and analyzed. Chi-square test was used to study the correlations between perinatal parameters and the morbidity rate of PIVH. Logistic regression analysis was used to detect the risk factor of PIVH. Results Ultrasonography of PIVH was characterized by hyperechoic masses in germinal matrix and ventricles, whereas in severe cases, ventricular dilatation and hyperechoic foci in the brain parenchyma could be scanned. Among 156 neonates, 49 cases(31.4%) were diagnosed with PIVH by CUS, including 45 cases(28.8%) of mild PIVH(grade Ⅰ and Ⅱ) and 4 cases(2.6%) of severe PIVH(grade Ⅲ and Ⅳ). There were statistically higher incidence of PIVH in neonates with low gestational age, low birth weight, 3 min Apagar score<7, respiratory distress syndrome, pneumonia and sepsis, and the differences were statistically significant(P<0.05). Logistic regression analysis showed that low gestational age was an independent risk factor for PIVH(OR=0.783, 95%CI: 0.639-0.959,P=0.018). Conclusion CUS is a reliable examination method to the diagnosis and grading of PIVH. CUS screening is essential to susceptible neonates with low gestational age, low birth weight and hypoxia related complications.

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Objective The study provided evidence for clinical prevention of colorectal cancer by analyzing the characteristics of recurrence after colorectal polypectom. Methods According to the number of polyps during the second colonoscopy, patients were divided into multiple recurrence group(≥3) and non-multiple recurrence group(<3). The characteristics and risk factors of polyp recurrence were analyzed. Results The clinical data of 370 patients who underwent two colorectal polypectomy were retrospectively analyzed. In univariate statistical analysis, polyp size(P=0.033), polyp site(P=0.002), and polyp color(P=0.032) were the risk factors for multiple polyp recurrence.In multivariate regression analysis, the number of polyps detected at the first colonoscopy(P=0.001) and the polyp site(P=0.001) were independent risk factors for multiple recurrence. Conclusion Multiple polyps on the first colonoscopy and polyps in the distal colon indicate an increased risk of multiple recurrence.

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Objective In the present study, the changes of left atrial matrix characteristics and the effect of catheter ablation on cardiac function in elderly patients with persistent atrial fibrillation were observed. Furthermore, the application of two catheter ablation methods including circular pulmonary vein isolation combined with left atrial matrix modification or left atrial posterior wall isolation were compared by researchers in order to explore effectiveness and safety of the two methods in elderly persistent atrial fibrillation patients. Methods A total of 86 elderly patients with persistent atrial fibrillation were selected and divided into two groups using randomization principle; the pulmonary vein isolation combined with left atrial matrix modified catheter ablation method was named matrix modified group(43 cases); the pulmonary vein isolation combined with left atrial posterior wall isolation catheter ablation method was named posterior wall isolation group(43 cases). During ablation left atrial electro matrix mapping were analyzed by multipole catheter for all patients. After ablation all patients were follow-up in 12 months by cardiac color doppler ultrasound examination, ECG and Holter. The incidence of arrhythmia, serious adverse event and complications were compared between two groups. Successful rate of single ablation and blank period recurrence after ablation were observed in 12 months follow-up time. Results Through electrogram mapping of left atrium low-voltage areas or scar areas were founded in 72 cases among the 86 cases after ablation, After a 12-month follow-up, the incidence of arrhythmia in the blank period between the two groups was 41.9%(matrix modified group) and 23.3%(posterior wall isolation group), independently,P<0.01, successful rates of single ablation in the two groups were 83.7%(matrix modified group), 60.5%(posterior wall isolation group), independently,P<0.01. No adverse event occurred in the two groups. After AF ablation LVEF increased, LVEDD and LVAD significantly decreased in all of the cases. There was no significant difference between the two groups in patients' characters ablation time and complications rate. Conclusion Circular pulmonary vein isolation combined with left atrial matrix modification is as safe as left atrial posterior wall isolation, and it is a simplified, personalized, and more effective ablation strategy in elderly patients with persistent atrial fibrillation.