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Objective To investigate the differential expression and distribution of forkhead-box O class 1(FoxO1) protein in lung cancer and esophageal carcinoma tissues, and to explore the association between subcellular distribution and function of FoxO1. Methods Immunohistochemistry was conducted to evaluate the cytoplasmic expression of FoxO1 in tissue microarray(TMA) of clinical lung cancer and esophageal carcinoma samples. The differential expression of FoxO1 in pattern and distribution between cancer and normal tissues were analyzed. Statistical analyses were performed to explore the correlation between the cytoplasmic FoxO1 protein and the clinicopathological characteristics of patients; the expression and localization of FoxO1 in lung cancer cell line A549 and esophageal carcinoma cell line Eca109 were analyzed by immunofluorescence; the siRNA knockdown and subsequent colony formation experiments were performed to investigate the effect of FoxO1 on tumor cell proliferation. Results The cytoplasmic FoxO1 expression in lung cancer and esophageal carcinoma tissues was higher than that in the corresponding paracancerous tissues(P<0.05), and there was no significance in the differential expression of cytoplasmic FoxO1 between lung squamous cell carcinoma and lung adenocarcinoma tissues; FoxO1 was dominantly localized to the cytoplasm, with less in the nucleus(P<0.05); the FoxO1 knockdown in A549 and Eca109 cells inhibited cell proliferation. Conclusion The expression of FoxO1 in lung cancer and esophageal carcinoma tissues is upregulated in the cytoplasm, while downregulated in the nucleus; the increased cytoplasmic expression of FoxO1 in cancer cells may promote cell proliferation.

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Objective To investigate the molecular mechanisms of inflammatory response and mucus hypersecretion of N-Acetyl-L-cysteine(NAC)or Resolvin-D1(RvD1)inhibiting hydrogen peroxide(H2O2)-induced diseased human bronchial epithelial cell of chronic obstructive pulmonary disease(COPD)primary airway epithelial cells(DHBE)of inflammation and mucus hypersecretion. Methods DHBE cells were isolated, identified and cultured. The oxidative stress model was established with the optimum stimulation concentration of H2O2by stimulating DHBE and incubated with NAC and RvD1in vitro. Reactive oxygen species(ROS) were measured by immunofluorescence and flow cytometry; IKKβ and NF-κB protein expressions and the phosphorylation level were detected respectively by Western blot; the mRNA expression and secretion levels of tumor necrosis factor(TNF)-α and interleukin(IL)-8 were detected by ELISA and qRT-PCR; mucin 5 AC(MUC5 AC) protein expression was detected by qRT-PCR and immunofluorescence. Results Compared with the control group, H2O2stimulated DHBE could significantly increase the production of ROS(P<0.05), the expressions of IKKβ and NF-κB phosphorylation increased in a concentration-dependent manner(P<0.05), The intracellular mRNA levels of TNF-α, IL-8 and protein levels of TNF-α, IL-8 in cell culture medium increased(P<0.05), MUC5 AC expression increased(P<0.05). NAC and RvD1 could significantly inhibit the expression of TNF-α, IL-8 and MUCSAC induced by H2O2(P<0.05). Conclusion NAC and RvD1 may be exerted protective effect against oxidative stress on inflammatory response and mucus hypersecretion of DHBE by the activation of the NF-κB pathway.

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Objective To study the effect of microRNA-200 a(miR200 a) on the progression of hepatocellular carcinoma and the therapeutic effect of sorafenib. Methods The stable transformation cell lines of human hepatocellular carcinoma miR200 a were constructed: miR200 a overexpression, Control and miR200 a knockdown. The expression of miR200 a in each transfected cell line was detected by qRT-PCR, the expression of protein was detected by Western blot method, the cell′s vitality was detected by MTT method, and the ability of cell migration and invasion was detected by transwell method. After adding sorafenib to the transfected cell line, the effect of miR200 a on the regulation of sorafenib on cells and the changes of cell vitality and corresponding protein expression were detected by MTT and Western blot. Eighteen SPF male BALB/c-nu mice were randomly divided into 3 groups, with 6 mice in each group. miR200 a overexpression, Control and miR200 a knockdown cells were subcutaneously implanted in each group. After tumor formation, each group of nude mice were injected with sorafenib every day for 14 days. The nude mice were killed and the tumor volume was calculated. The expression and localization of tumor target protein were verified by immunohistochemistry. Results miR200 a could inhibit the cell viability and migration ability of hepatocellular carcinoma cells, and reduce the expression of SIRT1 at the same time. MiR200 a significantly reduced the tumor volume of nude mice. Conclusion miR200 a can inhibit the proliferation and migration of human hepatocellular carcinoma cell line and increase the efficacy of sorafenib, which may be achieved by regulating the expression of SIRT1.

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Objective To investigate the influence of sodium aescinate on the autophagic flux in hepatocellular carcinoma(HCC), and the influence of chloroquine on the effect of sodium aescinate. Methods HCC cell lines(Huh7, HepG2 and Hep3B) were treated with sodium aescinate, then the proteins extracted from these cells were analyzed by the Western blot. After the co-administration of sodium aescinate and chloroquine, the Western blot, MTT assay, cell apoptosis assay, and colony formation assay were executed.Results Compared with the control, sodium aescinate upregulated the LC3B protein and inhibited the PI3K/AKT signaling pathway. The combination of sodium aescinate and chloroquine induced more expression of LC3B protein than the single drug. Furthermore, compared with the control or single drug, the combined treatment significantly decreased the cell viability(P<0.05), increased the early apoptosis(P<0.01), and attenuated the ability of colony formation(P<0.001).Conclusion Sodium aescinate induces the protective autophagy in HCC, and chloroquine can potentiate the effect of sodium aescinate.

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Objective To study the antibacterial effect of a new photosensitizer NHS-BODIPY-Br on the biofilm ofP gingivalisandT forsythia in vitro. Methods The biocompatibility of photosensitizer NHS-BODIPY-Br was detected by CCK8.P gingivalisandT forsythiawere cultured in anaerobic condition(10% CO2,10% H2,80% N2) for 48 h to form a biofilm and then treated with antibacterial photodynamic therapy.The photosensitizer concentration was 5,10,15 mg/L and the laser intensity was 15 and 30 J/cm2.After treatment, bacterial activity, residual LPS and laser confocal analysis were performed to determine the optimal treatment combination. Results The reduction rate ofbacterial biofilm was about 98%when the concentration of NHS-BODIPY-Br was 10 mg/L and the laser intensity was 15 J/cm2. Conclusion This study shows that the NHS-BODIPY-Br-mediated photodynamic therapy can effectively inhibit the biofilm ofP gingivalisandT forsythia.

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Objective To explore the effect of knocking down WD repeat domain 12(WDR12) on mouse spermatogonial cells(GC-1). Methods After transfecting cells with shWDR12 and negative control EGFP plasmids by liposome transfection, Western blot and Q-PCR were used to detect the expression of WDR12 in the cells; CCK-8 kit detected cell proliferation; TUNEL kit stains detected cell apoptosis; Q-PCR detected the differential expression of ribosomal protein genes in cells. Results Compared with the cells transfected with the negative control plasmid, the detection results of the cells transfected with shWDR12 showed that the expression of WDR12 in GC-1 cells was successfully reduced(P<0.01); the proliferation of GC-1 cells knocked down by WDR12 was inhibited; the results of TUNEL staining showed that the number of apoptotic cells increased after WDR12 knockdown; the expression of ribosomal protein genes decreased after shWDR12 transfection, ribosomal protein S3(RPS3)(P<0.05), ribosomal protein L11(RPL11)(P<0.01), ribosomal protein S15(RPS15)(P<0.05), and ribosomal protein L31(RPL31)(P>0.05). Conclusion Transfection of shWDR12 can effectively knock down the expression of WDR12 in GC-1 cells. The decrease of WDR12 expression leads to a decrease in the proliferation ability of GC-1 cells, an increase of apoptosis and the down-regulation of ribosomal-related protein gene expression.

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Objective To clarify the positive correlation between C-Myc and the expression of long non-coding RNA nuclear enriched abundant transcript 1(lncRNA NEAT1), and to explore how C-Myc regμlates lncRNA NEAT1 proliferation of oral quamous cell carcinoma(OSCC). Methods High-throμghput sequencing technology detected the expression of lncRNA in OSCC cells with knockdown of C-Myc, the expression of lncRNA NEAT1 in OSCC cells with overexpression and knockdown of C-Myc, overexpression and knockdown the proliferation ability of OSCC cells of lncRNA NEAT1; glycolysis stress detection experiment detected the glycolytic metabolism of OSCC cells overexpressing lncRNA NEAT1. Results The results showed that if C-Myc was overexpressed or knocked down, the expression of lncRNA NEAT1 showed an upward or downward trend. Overexpressing or knocking down lncRNA NEAT1, the proliferation ability of OSCC cells showed an upward or downward trend. Overexpression of lncRNA NEAT1 could increase the glycolysis level of OSCC cells. Conclusion C-Myc can increase the glycolysis level of OSCC cells by regulatinglncRNA NEAT1, and promote the proliferation of oral cancer cells and the development of oral cancer.

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Objective To explore the role of m6A demethylase FTO in the resistance of breast cancer cell to trastuzumab and the effect of the new anti-HER2 antibody HuA21 on the proliferation of drug-resistant breast cancer cell. Methods Trastuzumab-resistant cell line BT474/TR was prepared. The morphology of BT474/TR and BT474 cells were observed under optical microscope. MTT, immunofluorescence and flow cytometry were used to detect cell proliferation, the expression of proliferation-related antigen Ki67 and cell cycle. qPCR and Western blot methods were used to determine the expression of m6A-related enzymes FTO and METTL3. Results BT474/TR showed pyknosis of the nucleus, the pseudopodia disappeared, and the cell proliferation rate decreased. As BT474 transited to BT474/TR with trastuzumab treatment, Ki67 expression and S phase cell proportion gradually decreased(P<0.05), and FTO expression gradually increased(P<0.05). However, there was no difference in METTL3 expression. The FTO inhibitor MA2 promoted the BT474/TR proliferation and increased its sensitivity to trastuzumab treatment(P<0.05). HuA21 antibody had an inhibitory effect on the proliferation of BT474/TR cells(P<0.05), and the combination of MA2 and trastuzumab had a better inhibitory effect on BT474/TR cells(P<0.05). Conclusion There is a correlation between the resistance of HER2-positive breast cancer cell to trastuzumab and FTO gene expression and the inhibition of FTO activity can enhance the sensitivity of drug-resistant cells to trastuzumab treatment. Moreover, HuA21 antibody has an inhibitory effect on the proliferation of BT474/TR cells.

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Objective To investigate the protective effect of ulinastatin(UTI) on the myocardial hypertrophy induced by isoproterenol in rats and the possible mechanism. Methods Rat model of myocardial hypertrophy was established by subcutaneous injection of 5 mg/(kg·d) of isoproterenol for seven consecutive days. Ultrasound examination was used to detect cardiac function and myocardial hypertrophy; heart mass index(HMI) and left ventricular mass index(LVMI) were measured. Cardiac function and myocardial hypertrophy were measured by ultrasound. Myocardial tissue was examined by HE staining and Masson staining. Western blot was used to detect the expressions of autophagy protein Beclin-1, ATG-5, ATG-7, LC3-Ⅱ autophagy protein as well as inflammatory factor tumor necrosis factors(TNF)-α and interleukin(IL)-6. Results Compared with the ISO group, diastolic ventricular septal thickness, left ventricular posterior wall depth as well as HMI and LVMI in the UTI group significantly decreased, while left ventricular ejection fraction and left ventricular short axis shortening rate significantly increased; histopathological examination showed that UTI could significantly inhibit isoproterenol-induced myocardial injury and myocardial fibrosis; UTI could also significantly inhibit isoproterenol-increased protein expressions of TNF-α,IL-6 and autophagy-associated protein Beclin-1, ATG-5, ATG-7, LC3-Ⅱ in rat myocardium. Conclusion UTI significantly improves isoproterenol-induced cardiac dysfunction and myocardial hypertrophy in rats, which may be related to the inhibition of inflammation and autophagy.

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Objective To explore the relationship between the expression of macrophage-derived Bruton′s tyrosine kinase(BTK) with renal function in the kidneys of patients with diabetic nephropathy(DN). Methods 49 patients with DN diagnosed by renal biopsy were collected as the DN group. 18 normal human serum and 18 normal kidney tissues adjacent to the cancer in patients with kidney cancer were collected as a control group. Clinical indicators of all participants were collected. The expression of interleukin(IL)-1β, tumor necrosis factor(TNF)-α and monocyte chemotactic protein(MCP)-1 in the serum of the control group and the DN group was detected by ELISA. The expression of TNF-α, CD68, BTK and p-BTK in renal tissues of the control group and the DN group was detected by immunohistochemistry. Laser confocal was performed to observe the co-expression of inducible carbon monoxide synthase(iNOS) and p-BTK in kidney tissue macrophages in the control group and the DN group.Results Compared with the control group,the levels of Hb A1 c,blood pressure,creatinine,urea nitrogen and uric acid in DN group significantly increased,and the levels of serum inflammatory cytokines TNF-α,IL-1β and MCP-1 significantly increased. Compared with the control group,the expression of TNF-α,the infiltration of macrophages and the activation of BTK significantly increased in DN group,and the expressions of iN OS and p-BTK in renal macrophages also increased. In DN group,BTK and p-BTK expression were positively correlated with 24 hour urinary protein,serum creatinine and blood urea nitrogen,and negatively correlated with estimated glomerular filtration rate( e GFR). Conclusion The activation of BTK in renal macrophages is increased in DN patients and is correlated with renal function progression.

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Objective To explore the preventive effect of epigallocatechin gallate(EGCG) on dental caries through the rat dental caries model, and provide experimental and theoretical basis for clinical research of natural medicine EGCG on preventing caries. Methods Twenty-four SD rats were selected and randomly divided into EGCG group, sodium fluoride(NaF) group, chlorhexidine acetate(CHX) group and distilled water(control) group, each with 6 rats. The rats were inoculated with standard strain ofstreptococcus mutans(S.mutans) and given cariogenic feed 2000# and 5% sucrose solution. The oral cavity of rats was rinsed and coated with minimum inhibitory concentration(MIC) EGCG, 250 mg/L NaF solution, 0.12% CHX solution and distilled water, respectively. After 6 weeks of continuous treatment, the rats were sacrificed. The jaws of the rats were collected and analyzed by Keyes score method to evaluate the caries of the rats′ maxillary molars. Results The Keyes scores of caries showed that compared with the control group, drugs in each group had different degrees of inhibition effect on the formation and severity of caries in rats. Among them, the EGCG group had better effect on enamel caries(P<0.01) and dentin caries, and had much better effect on dentin deep caries(P<0.001) of maxillary fossa and fissure caries in rats. However, the prevention and treatment effect of superficial and middle dental caries was lower than that of CHX group. Conclusion EGCG with MIC concentration reduces the incidence and severity of dental caries, and has a significant inhibitory effect on the occurrence of dental caries in rats.

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Objective To investigate the expression of sigma-1 receptor in oral squamous cell carcinoma(OSCC) tissues and cell lines, reduce the expression of sigma-1 receptor in OSCC cells by transfection with specific siRNA, detect the effect on their biological behavior, and preliminarily analyze the possible mechanism. Methods The expression of sigma-1 receptor in human OSCC tissues were detected by immunohistochemistry. Western blot and immunofluorescence were used to detect the expression of sigma-1 receptor in human OSCC cell lines. GEPIA database was used to analyze the correlation between sigma-1 receptor expression level and prognosis of patients withOSCC. The expression of sigma-1 receptor was down-regulated by transfection of specific siRNA. Then the proliferation, migration and invasion of OSCC cells after transfection were detected by CCK-8 and Transwell assay, and the expression level of voltage-gated sodium ion channels subtype Nav1.5 in OSCC cells was detected by qRT-PCR and Western blot. Results Immunohistochemical analysis showed that the expression of sigma-1 receptor was higher in OSCC tissues than that in normal tissues. Western blot revealed that the sigma-1 receptor was expressed in all four OSCC cell lines. The results of GEPIA database analysis showed that the expression level of sigma-1 receptor was negatively correlated with the overall survival rate of patients with OSCC. Immunocytochemistry and confocal microscopy showed that the sigma-1 receptor was mainly localized in the plasma membrane in CAL-27 cells. CCK-8 analysis showed that the down-regulated expression of sigma-1 receptor reduced the proliferation of CAL-27 cells through its specific siRNA. Transwell analysis showed that the reduced expression of sigma-1 receptor significantly inhibited the migration and invasion of CAL-27 cells. In addition, down-regulation of sigma-1 receptor expression was found to inhibit the protein and mRNA expression levels of Nav1.5. Conclusion sigma-1 receptor participates in the occurrence and development of OSCC, promotes the proliferation, migration and invasion of OSCC cells, and may be involved in the progression of OSCC by regulating the Nav1.5.

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Objective To investigate the effect of SIRT6 on the proliferation and migration of renal cancer cell lines ACHN and 786-O. Methods Immunohistochemistry was used to detect the expression of SIRT6 in renal cell carcinoma and adjacent tissues; Western blot was used to detect the expression levels of kidney cancer cell line SIRT6 and related downstream signaling pathway molecules; SIRT6-siRNA transfection and SIRT6 specific inhibitor OSS_128167 were used to treat ACHN cells, Scratch test and Tetramethylazazole blue test were usedto observe the proliferation and migration ability of treated ACHN cells, and Western blot was used to detect changes in downstream related signal pathways. Results The results of immunohistochemistry and Western blot showed that the expression of SIRT6 in renal cancer tissues was significantly higher than that in adjacent tissues. Compared with normal renal tubular epithelial cells HK-2, the expression of SIRT6 in renal cancer cell line ACHN significantly increased. After transfection of ACHN with SIRT6-siRNA, the expression of SIRT6 was significantly reduced. Scratch test and MTT showed that the proliferation and migration ability of ACHN cells was reduced. After OSS_128167 treatment of renal cancer cell lines, the deacetylation activity of SIRT6 decreased, and the cell proliferation ability decreased. Conclusion The SIRT6 deacetylase can inhibit the proliferation and migration of renal cancer cells by down-regulating the PI3 K-AKT pathway, and may become a potential new target for drug therapy.

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Objective To investigate the effects of up-regulation of GADD45α by Lentivirus vector on the proliferation, migration and apoptosis of imatinib-resistant K562 IR cells. Methods K562 cells and imatinib-resistant K562 IR cells were cultured. The expression levels of GADD45α mRNA and protein in K562 and K562 IR cells were detected by qRT-PCR and Western blot. The Lentivirus vector that over-expressed GADD45α and control were transfected in K562 IR cells, respectively Lv-GADD45α and Lv-NC. The expression level of GADD45α in each group was detected by Western blot, and the changes in cell proliferation, migration and apoptosis after transfection were further detected by CCK-8, Tanswell and Annexin V-FITC/PI double staining. Results The expression levels of GADD45α protein and mRNA in K562 IR cells were lower than those in K562 cells. After GADD45α was transfected, the expression levels of GADD45α in Lv-GADD45α group were higher than those in Lv-NC group. After imatinib treatment, the IC50value of Lv-GADD45α group was lower than that of Lv-NC group, the apoptosis rate of cells in Lv-GADD45α group was higher than that of the Lv-NC group, and the cell migration ability also decreased, with statistically significant difference(P<0.05). Conclusion By up-regulating GADD45α in imatinib-resistant K562 IR cells, the sensitivity of K562 IR to imatinib in chronic myeloid leukemia cells is enhanced, leading to proliferation inhibition and the apoptosis of chronic myeloid leukemia cells.

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Objective To explore the effect of ATP-binding cassette transporter subfamily G2(ABCG2) gene on human gastric cancer cells and explore its possible mechanism. Methods 84 pathological specimens that are positive for epidermal growth factor 2(HER2) were collected. ABCG2 protein expression was detected by immunohistochemistry, and their clinicopathological characteristics was analyzed. Lentivirus was transfected into HER2-positive gastric cancer cell lines NCI-N87 and MKN-7 to construct a stable cell line model; RT-PCR, Western blot and immunofluorescence were used to detect transfection efficiency and ABCG2 expression; Scratch experiment was used to observe the influence of ABCG2 on cell migration; Western blot was used to detect the protein expression of HER2, PI3 K/Akt signaling pathway. Results Among 84 pathological specimens, 60 cases were ABCG2 positive(71.4%) while 24 cases were negative(28.6%). The double positive of HER2 and ABCG2 was closely related to the differentiation degree of gastric cancer and the later treatment; compared with the Cell group and the LV5 NC group, the mRNA and protein expression levels of ABCG2 in the LV5-homo group increased(P<0.01). Compared with the Cell group and the LV5 NC group, the expression levels of HER2, PI3 K and Akt phosphorylation(P-Akt) in the signaling pathway of the LV5-homo group increased(P<0.05). Compared with the Cell group and the LV5 NC group, the expression of ABCG2 in the LV5-homo group increased and promoted the migration of gastric cancer cells(P<0.05). Conclusion ABCG2 is widely expressed in HER2-positive gastric cancer cells and is closely related to the metastasis, differentiation and TNM staging of gastric cancer; ABCG2 up-regulates the HER2-PI3 K/Akt pathway, promotes the migration of gastric cancer cells, and affects the expression of key proteins in the pathway, which provides a new idea for the study of drug resistance mechanisms in gastric cancer.

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Objective To explore the influences of activating transcription factor 4(ATF4) on the proliferation, migration and apoptosis of hepatocellular carcinoma(HCC) and itsmolecular mechanism.Methods Expression of ATF4 andthe hexokinase domain component 1(HKDC1) were down-regulated in HepG2 and Huh7 cells by RNA interference technique. In HCC cell lines, CCK-8, clone formation, wound-healing assay and flow cytometry apoptotic analysis were used to observe the changes in the proliferation, migration and apoptosis of HCC after transfection, and the expressions of ATF4,HKDC1 and apoptosis-related proteins Bcl-2 and Bax in hepatocellular carcinoma cells were determined by Western blot. Results Functional studies had shown that knockdown of ATF4 and HKDC1 not only led to an increase in cell apoptosis and a decrease in cell proliferation but also inhibited the cells' abilities of migration; ATF4 silencing could inhibit the expression of HKDC1 protein. Conclusion ATF4 may play a role in the progression of hepatocellular carcinoma by regulating the expression of HKDC1 protein,and HKDC1 may be a potential target of therapeutic intervention for HCC.

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Objective To compare the early osteogenesis effect of three commercialized implant surfaces SLA®,SLActive®,Xpeed®in maxillary sinus.Methods The adhesion and proliferation of MC3T3E1 cells on three titanium disks were evaluated by cell staining and CCK-8. In vivo study, 16 New Zealand rabbits were randomly divided into SLActive group and Xpeed group. Bilateral maxillary sinus external augmentation model was established. The SLA discks were used as control group, and the rest discks were implanted according to preceding group. The mRNA expression of bone morphogenetic protein 2(BMP2), core binding factor(Runx2), osteocalcin(OCN), type I collagen(COLI) were measured by qRT-PCR in 4 weeks after surgery. To evaluate their osteogenic effect, Micro CT was used to analyze the bone miner density(BMD), trabecular thickness(Tb.Th), trabecular separation(Tb.Sp) and trabecular number(Tb.N) of disk. Results The result of 6-hour cell adhesion showed no statistically significant difference in three disks, and 24-hour fluorescent staining showed that the cell morphology and spread in the Xpeed group were more sufficient than those in SLA and SLActive groups. CCK-8 test showed that cell proliferation in slactive group and Xpeed group on day 3 and 7 was higher than that in SLA group(P<0.05), there was no statistical difference between the SLActive and Xpeed groups. qRT-PCR showed that osteogenic related gene expression level in Xpeed group was higher than that in SLA and SLActive groups(P<0.01). The results of micro-CT showed that the Tb.Th, Tb.Sp of each group showed statistically significant differences(P<0.05), Tb.Th of Xpeed was larger than that of SLActive and SLA group.Tb.Sp in Xpeed was less than that in SLActive group and SLA group(P<0.05).Conclusion Which shows that Xpeed®surface has a better effect on early osteogenesis in maxillary sinus.

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Objective To observe the effects and mechanisms of plasma-activated solution(PAS) on the proliferation and apoptosis of hemangioma endothelial cells(HemECs). Methods Phosphate buffer saline(PBS) was treated by cold atmospheric plasma(CAP) for 10 s,20 s,30 s,40 s and 50 s to form PAS. The PBS with 0 sCAP treatment was taken as the control group. The p H value and active medium concentration of PAS was detected by electronic p H meter and kit respectively. CCK-8 kit was used to detect cell proliferation. Flow cytometry was used to detect cell apoptosis. Western blot was used to detect the expression of kinase B(AKT),phosphorylated AKT(pAKT),phosphatidylinositol-3 kinase(PI3K) and phosphorylated PI3K(p-PI3K) protein in the HemE Cs. Results Compared with the control group,the p H value of PAS gradually decreased in a dose-dependent manner. And the concentration of active mediums such as NO3-,H2O2and O3in PAS increased in a dose-dependent manner.Compared with the control group,PAS significantly increased the level of ROS in HemE Cs,inhibited the proliferation and promoted the apoptosis. Compared with the control group,the expression of p-PI3K had no significant change,and the expression of p-AKT had no significant change at 10 s. However,the expression of p-AKT decreased rapidly,when the processing time increased to 20 s,30 s,40 s,and 50 s. Conclusion PAS can inhibit proliferation and promote apoptosis of HemE Cs. This effect is related to the increase of intracellular ROS level and the down-regulation of AKT phosphorylation level.

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Objective To investigate the effect of miR-223-3 p on osteogenic differentiation of human dental pulp stem cells(hDPSCs) by inhibiting Von Hippel-Lindau(VHL) gene. Methods Hypoxia induced hDPSCs for 24 h(oxygen concentration 1%).The changes of cell proliferation were observed by MTT assay.The miRNA of miR-223-3 p was predicted by bioinformatics, and the relationship between miR-223-3 p and VHL was verified by luciferase.The expression of miR-223-3 p mimics or miR-223-3 p was transfected into DPSCs, and the expression of miR-223-3 p, VHL, hypoxia inducible factor-1α(HIF-1α),runt-related transcription factor 2(Runx2), bone morphogenetic protein 2(BMP2) mRNA, VHL, HIF-1α was detected by qRT-PCR and Western blot. Results miR-223-3 p increased in the hypoxic-induced DPSCs(P<0.05).The MTT results showed that miR-223-3 p mimics promoted the proliferation of DPSCs(P<0.05).Moreover, miR-223-3 p regulated the expression level of VHL by targeting VHL 3′-UTR.The transfection of miR-223-3 p was detected by qRT-PCR and Western blot, which significantly promoted the expression of HIF-1α, Runx2 and BMP2 genes(P<0.05). Conclusion miR-223-3 p promotes osteogenic differentiation of dental pulp stem cells by inhibiting VHL gene.

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Objective To study the protective effect and molecular mechanism of long non-coding RNA(lncRNA) PWAR5 on cardiomyocytes in lipopolysaccharide-induced myocarditis. Methods The primary rat cardiomyocytes were isolated and cultured, and were infected with lentivirus carrying PWAR5 sequence(PWAR5 group) and empty lentivirus(Control group). Real-time quantitative polymerase chain reaction(qRT-PCR) was used to detect infection efficiency. After lipopolysaccharide-induced myocarditis, the MTS method was used to detect the effect of PWAR5 on the viability of primary rat cardiomyocytes, and the ELISA method was used to detect the levels of interleukin(IL)-1β and tumor necrosis factor(TNF)-α in the supernatant. Flow cytometry was used to detect the effect of PWAR5 on the apoptosis of primary rat cardiomyocytes. Bioinformatics methods and dual-luciferase reporter gene experiments were used to predict and verify the target gene of PWAR5. qRT-PCR was used to detect the expression of target gene. Western blot was used to detect the expression of target gene protein and apoptosis-related proteins. Results Compared with the Control group, the expression of PWAR5 in the PWAR5 group increased(P<0.01). After lipopolysaccharide treatment, compared with the Control group, the cell viability of the PWAR5 group increased(P<0.01), the contents of IL-1β and TNF-α in the supernatant decreased(P<0.01), and the percentage of apoptosis in the PWAR5 group decreased(P<0.01). The target gene of PWAR5 might be miR-30 a-5 p, and PWAR5 could complement miR-30 a-5 p(P<0.01). Compared with the Control group, the expression of miR-30 a-5 p in the cells of the PWAR5 group decreased(P<0.01), and the expression of suppressor of cytokine signaling 3(SOCS3) mRNA increased(P<0.01). Compared with the Control group, the expression of SOCS3 and Bcl-2 protein in the PWAR5 group increased, while the expression of Bax and Caspase-3 protein decreased. Conclusion PWAR5 can reduce the damage of cardiomyocytes caused by myocarditis by targeting the miR-30 a-5 p/SOCS3 molecular axis.

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Objective To explore the relationship between weight change since 20 years old and hyperuricemia(HUA) in a middle-aged Hefei population. Methods The subjects were 1 727 adults aged 45 to 60. Data was collected through face-to-face questionnaire and physical examination. The relationship between weight change since 20 year and HUA in a middle-aged Hefei population was studied by multivariate logistic regression analysis. Results After adjusting for age, education, smoking and other confounding factors, compared with men who gained<5 kg, the risk of HUA in men who gained 5-10 kg, ≥10 kg significantly increased(OR=2.12, 95%CI: 1.09-4.13,OR=2.82, 95%CI: 1.61-4.91). Compared with women who gained<10 kg, the risk of HUA in women who gained≥10 kg also significantly increased(OR=3.44, 95%CI: 1.66-7.13). Conclusion Weight gain in adulthood is associated with an increased risk of HUA, especially in men.

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Objective To investigate the correlation between the cerebral gray-matter volume changes and Montreal Cognitive Assessment Scale(MoCA), cerebral structure of end-stage renal disease(ESRD) is evaluated by using voxel-based morphometry(VBM). Methods 47 patients with end-stage renal disease(ESRD group) and 46 normal volunteers(control group) underwent T1 WI structure scanning and MoCA tests. The DPABI software was used to preprocess the cerebral structure data, independent samplet-test was performed for cerebral structure data of two groups, and the difference of brain regions was obtained by using SPM 12(P<0.05,FWE corrected). Pearson correlations between volume of brain regions of difference and MoCA score were conducted by using SPSS(P<0.05). Results Compared with healthy control group, gray-matter volume of ESRD group decreased in the left caudate, left parahippocampal gyrus, left anterior cingulate gyrus, left transverse temporal gyrus, left middle frontal gyrus, left anterior insula, bilateral middle cingulate gyrus, right temporal pole and right precentral gyrus medial segment(allP<0.05,FWE corrected). The volume of the left anterior cingulate gyrus positively was correlated with MoCA score, attention, language and abstract score(P<0.05), that of the left anterior insula positively was correlated with MoCA score, visual and executive ability, attention score(P<0.05), that of the left middle cingulate gyrus positively was correlated with attention score(P<0.05), and that of the right precentral gyrus medial segment positively was correlated with language score(P<0.05). Conclusion The volume of some brain regions of ESRD patients decreases, involving left anterior/middle cingulate gyrus, left anterior insula and right precentral gyrus medial segment. The brain regions are mainly related to cognitive, attention and language.

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Objective To explore the mechanism of positive Hoffman sign in patients with cervical spondylosis myelopathy(CSM). Methods DTI analysis was performed in 88 patients with CSM(50 positive and 38 negative with Hoffman sign) and 30 healthy controls. In the high cervical spinal cord level(C1-C3), the reticular spinal tract, thin wedge tract, anterior corticospinal tract and lateral tract were selected as the regions of interest, and it was analyzed whether the DTI parameters of different conduction tract areas within and between groups were statistically significant. Results There was a significant difference in the ratio of DTI parameters in different conduction tract areas of the high cervical spinal cord in patients with CSM. Compared with the Hoffman sign negative group and the healthy control group, the Hoffman sign positive group showed lower FA ratio and higher ADC ratio in the corticospinal tract area(P<0.05). Conclusion The occurrence of Hoffman sign is related to the damage of corticospinal tract. For CSM patients with positive Hoffman sign, the microstructural damage of the spinal cord has occurred at the high level of the cervical spinal cord far from the lesion.

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Objective To investigate the prevalence of sarcopenia in patients with lung cancer, and to explore the influencing factor in lung cancer patients with sarcopenia. Methods A total of 78 lung cancer patients(lung cancer group) and 78 healthy subjects(control group) were selected. The body measurement of the two groups were measured by bioelectrical impedance method. Height-adjusted appendicular skeletal muscle mass index(ASMI) was used to define sarcopenia. Results The prevalence of sarcopenia in lung cancer patients was higher than that in controls(61.5%vs17.9%, χ2=30.943,P<0.001). Compared with the control group, the protein, fat free mass(FFM), soft lean mass(SLM), appendicular skeletal muscle mass(ASM) and ASMI levels of the lung cancer group decreased(allP<0.05), while the visceral fat area(VFA) of lung cancer group increased(P<0.001). The weight, body mass index(BMI), protein, FFM, SLM, ASM, ASMI and body fat mass(BFM) of lung cancer patients with sarcopenia were lower than those of lung cancer patients without sarcopenia(allP<0.05). There was a difference in the incidence of sarcopenia between different BMI groups in the lung cancer patients(100%vs62.5%vs33.3%, χ2=13.569,P=0.001). Pearson correlation analysis showed that ASMI was positively correlated with height, weight, BMI, protein, FFM, SLM and ASM(allP<0.001), while it was negatively correlated with age and percent body fat(PBF)(allP<0.05). Binary Logistic analysis showed that higher BMI(OR=-1.997, 95%CI: 0.046-0.400,P<0.001) and higher ASM(OR=-2.696, 95%CI: 0.012-0.389,P=0.003) were the independent protective factors for the lung cancer patients with sarcopenia. Conclusion Lung cancer patients have higher prevalence of sarcopenia. Higher BMI and higher ASM are the protective factors for the incidence of sarcopenia in patients with lung cancer.

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Objective To investigate the immune exhaustion of CD4+T cells and CD8+T cells in peripheral blood of patients with hepatocellular carcinoma(HCC) and its relationship with disease progression. Methods 30 patients with HCC and 20 healthy subjects(HC) were collected. Flow cytometry was used to detect the frequencies CD4+T cells and CD8+T cell expressing surface inhibitory receptors PD-1, TIGIT, TIM-3 and intracellular IL-10. Cell microsphere array(CBA) was used to detect the content of serum cytokine(IL-10). Results The percentages of TIM-3+CD4+T cells, TIGIT+CD4+T cells, PD-1+CD4+T cells, TIM-3+TIGIT+CD8+T cells, IL-10+CD4+T cells, TIM-3+TIGIT+CD4+T cells in peripheral blood of HCC patients was higher than those of healthy control group(P<0.001), the percentages of TIM-3+CD8+T cells, TIGIT+CD8+T cells and PD-1+CD8+T cells in peripheral blood of HCC patients were higher than those in healthy control group(P<0.01). In addition, the proportions of PD-1+TIM-3+TIGIT+CD4+T cells and PD-1+TIM-3+TIGIT+CD8+T cells in HCC patients were respectively higher than those in PD-1+TIM-3-TIGIT-CD4+T cells and PD-1+TIM-3-TIGIT-CD8+T cells(P<0.001), and serum IL-10 levels in HCC patients were higher than that in healthy controls(P<0.05). ROC curve analysis showed that the frequencies of TIM-3+TIGIT+CD8+T cells and TIM-3+TIGIT+CD4+T cells in the diagnosis of HCC was higher than the percentage of mono-molecule positive cells. The neutrophil/lymphocyte ratio(NLR) of the groups with high TIM-3+TIGIT+CD8+T cells and high TIM-3+TIGIT+CD4+T cells ratio were higher than those of the groups with low TIM-3+TIGIT+CD8+T cells and low TIM-3+TIGIT+CD4+T cells ratio group(P<0.05). Conclusion The proportion of TIM-3+TIGIT+CD4+T cells and TIM-3+TIGIT+CD8+T cells in peripheral blood of HCC patients is higher than that of the control group, and PD-1 is highly expressed in all above cells, which is closely related to IL-10 concentration. And high percentages of TIM-3+TIGIT+CD4+T cell and TIM-3+TIGIT+CD8+T cell are associated with HCC progression disease and poor prognosis, indicating that double molecular targeted therapy may have greater clinical significance than single molecule targeted therapy. Therefore, this work may provide a new and more effective therapeutic target for HCC according to the bimolecular targeted therapy, and promote the development of immune checkpoint targeted therapy for HCC patients.

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Objective To study the expression and clinical significance of annexin A1(ANXA1) in papillary thyroid carcinoma(PTC) and explore the role of ANXA1 in the development of PTC and its relative molecular mechanism. Methods From the TCGA public database, the relevant data of ANXA1 expression was obtained and analyzed in thyroid cancer and adjacent normal tissues, and explored the relationship between the ANXA1 expression and the patient′s overall survival; then the paraffin sections were collected which contained PTC, nodular goiter(NG), and normal thyroid tissue(NT), and the expression of ANXA1 protein were detected by immunohistochemical method; the fresh tissues were collected and ANXA1 mRNA was checked by the way of qRT-PCR; then the relationship between the expression level of ANXA1 and clinicopathological characteristics was studied. Results Through database analysis, the expression of ANXA1 in thyroid cancer was more prominent than that in NT(P<0.05), and the expression level of ANXA1 was associated with the survival of patients(P<0.05); ANXA1 protein had a higher level in the paraffin sections of PTC, which compared with NT and NG; and ANXA1 mRNA was lower than them. The results of clinical data analysis showed that ANXA1 was related to the location, diameter and lymph node metastasis(P<0.05), but had nothing with the patient′s age, gender and clinical stage. Conclusion ANXA1 has a closed relationship with PTC, which is essential to potential diagnostic and prognostic indicators.

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Objective To analyze the value of accruate quantitative SPECT/CT in differential diagnosis of benign and malignant thyroid nodules. Methods 81 patients with thyroid nodules diagnosed by ultrasound were selected.All patients were examined by99mTcO4-imaging and quantitative SPECT/CT,cool and cold nodules were performed99mTc-MIBI imaging and quantitative SPECT/CT on the next day. Region of interest( ROI) was used to obtain thyroid uptake rate and nodule's standard uptake value. The difference between the parameters of benign and malignant nodules was compared based on patient's postoperative pathology. The diagnostic thresholds of standardized uptake value( SUV) were obtained by drawing the ROC curves,and the diagnostic efficacy was evaluated by comparing with the results of99mTcO4-and99mTc-MIBI imaging. Results In 81 patients,postoperative pathology showed that 35 thyroid nodules were malignantand 46 thyroid nodules were benign. The difference of thyroid uptake rate and nodule's standard uptake value between benign and malignant groups was statistically significant( P<0. 05).The diagnostic sensitivity,specificity and accuracy of99mTcO4-combined with99mTc-MIBI imaging were 41. 6%,68. 8%,56. 8% respectively. ROC curve results showed the diagnostic thresholds,area under curve( AUC),sensitivity,specificity,accuracy of99mTcO4-SUVmax and SUVavg were 43. 83 g/ml,0. 808,100%,51. 1%,72. 8% and30. 08 g/ml,0. 808,94. 1%,57. 4%,74. 1% respectively. The diagnostic thresholds,AUC,sensitivity,specificity,accuracy of99mTc-MIBI SUVmax and SUVavg were 2. 24 g/ml,0. 899,79. 4%,92. 9%,85. 4% and 1. 37 g/ml,0. 816,88. 2%,71. 4%,82. 2% respectively. Conclusion Accurate quantitative SPECT/CT has high accuracy in diagnosis of benign and malignant thyroid nodules,and99mTc-MIBI SUVmax has the highest diagnostic efficiency when the diagnostic threshold is 2. 24 g/ml.

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Objective To analyze the results of the micro lymphotoxin cross-matching test and explore the primary distribution of donors and recipients and its clinical significance. Methods 344 patients were analyzed, including age, gender, blood group distribution, donor-recipient relationship, and the influence was also investigated, respectively. Results In the aspect of recipients: the number of male recipients was more than that of female recipients, and the age of recipients was mainly between 21 and 50 years old. The PDL of 31～40 years receivers was the lowest in the low-risk group, which was(3.85 ±2.08); the recipients′ blood types were mostly typed A and O, of which 85.17% of the donors and recipients had the same blood type. In the aspect of donors: 72.67% of the donors were DCD;in the low-risk group, the PDL of the DCD group was(5.30±2.24), which was higher than that of LDR group, and the difference was statistically significant(P<0.05). In LDR, female donors were more than male donors, the donors over 50 years old were the most, type O donors were the most, and 93.02% of the donors were parents. Conclusion Although the number of DCD was more than that of LDR, the positive rate of PDL was lower in LDR, and it can provide kidney sources more quickly and effectively solve the pain of patients with renal failure.

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To investigate the clinical characteristics and lymphocyte subsets of children with infectious mononucleosis complicated with multiple pathogens. One hundredchildren with IM were included and divided into simple EB virus infection group(53 cases) and multiple infection group(47 cases) according to pathogen infection. In addition,80 healthy children in the outpatient department of pediatrics were selected as the control group. The distribution characteristics of pathogens in the multiple infection group were analyzed. The clinical characteristics of simple EBV infection group and multiple infection group were compared,and the changes of lymphocyte subsets among three groups were compared. The main co-infected pathogens in children with IM were Mycoplasma pneumoniae(MP)(53. 19%) and Cytomegalovirus(CMV)(42. 55%). The incidence of pharyngitis,splenomegaly and liver function damage in the multiple infection group was significantly higher than that in the simple EBV infection group(P<0. 01). The expression levels of CD8+and CD3+in multiple infection group and simple EBV infection group were significantly higher than those in the control group(P<0. 01),while CD4+and CD4+/CD8+significantly decreased(P<0. 01). There were also differences between multiple infection group and simple EBV infection group(P<0. 01). The distribution characteristics of pathogens in children with IM were diverse,and the multiple infectious of EBV,MP and CMV were the main ones. Compared with simple EBV infection,the clinical symptoms of children with multiple infections were more serious,lymphocyte subsets were more disordered,and immune function was lower.

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To analyze the clinical data of 362 patients with acute cerebral infarction who underwent mechanical thrombectomy. Cardiogenic embolism was the most common type according to the trial of org 10172 in acute ischemic stroke test(TOAST) classification. The internal carotid system was the most frequently involved. According to the modified rankin score(mRS), there were 157 patients in good prognosis group(mRS≤2 score) and 205 patients in poor prognosis group(mRS≥3 score). There were significant differences in age, hypertension, vascular involvement, national institutes of health stroke scale(NIHSS) score at admission and discharge, postoperative pulmonary infection and intracranial hemorrhage between the two groups(P<0.05). The risk factors for poor prognosis were advanced age, involvement of vertebrobasilar artery system, high NIHSS score at discharge and postoperative pulmonary infection.