T cell infiltration and proliferation in tumor tissues are the main factors that significantly affect the therapeutic outcomes of cancer immunotherapy. Emerging evidence has shown that interferon-gamma (IFNγ) could enhance CXCL9 secretion from macrophages to recruit T cells, but Siglec15 expressed on TAMs can attenuate T cell proliferation. Therefore, targeted regulation of macrophage function could be a promising strategy to enhance cancer immunotherapy via concurrently promoting the infiltration and proliferation of T cells in tumor tissues. We herein developed reduction-responsive nanoparticles (NPs) made with poly (disulfide amide) (PDSA) and lipid-poly (ethylene glycol) (lipid-PEG) for systemic delivery of Siglec15 siRNA (siSiglec15) and IFNγ for enhanced cancer immunotherapy. After intravenous administration, these cargo-loaded could highly accumulate in the tumor tissues and be efficiently internalized by tumor-associated macrophages (TAMs). With the highly concentrated glutathione (GSH) in the cytoplasm to destroy the nanostructure, the loaded IFNγ and siSiglec15 could be rapidly released, which could respectively repolarize macrophage phenotype to enhance CXCL9 secretion for T cell infiltration and silence Siglec15 expression to promote T cell proliferation, leading to significant inhibition of hepatocellular carcinoma (HCC) growth when combining with the immune checkpoint inhibitor. The strategy developed herein could be used as an effective tool to enhance cancer immunotherapy.

Objective To investigate the mechanism of action of integrin α4 (ITGA4) in liver fibrosis based on the anti-liver fibrosis effect of sticky sugar amino acid (SSAA) in rats. Methods A rat model of liver fibrosis was induced by intraperitoneal injection of CCl 4 , and then colchicine and low-, middle-, and high-dose SSAA were used for intervention, with blank control group and SSAA group as control. After 12 weeks of experimental intervention, serum and liver samples were collected to measure the serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and HE staining and Sirius Red staining were used to observe the pathological conditions of liver tissue; quantitative real-time PCR was used to measure the transcriptional level of ITGA4, integrin β1 (ITGB1), transforming growth factor-β1 (TGFβ1), alpha-smooth muscle actin (α-SMA), and TIMP2 in liver tissue; Western blot was used to measure the relative protein expression levels of ITGA4, ITGB1, TGFβ1, α-SMA, MMP2, TIMP1, and TIMP2; immunohistochemistry was used to observe the protein expression of TGFβ1 and α-SMA. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for comparison between two groups. Results There were significant increases in AST and ALT in the CCl 4 model group, and intervention with colchicine or low-, middle-, and high-dose SSAA reduced the levels of AST and ALT, with a significant difference between the CCl 4 model group and the other groups (all P < 0.05). HE staining and Sirius Red staining showed disordered structure of hepatic lobules and an increase in collagen fibers in the CCl 4 model group, and the structure of hepatic lobules was improved after intervention with colchicine or low-, middle-, and high-dose SSAA. The CCl 4 model group had significantly higher transcriptional levels of ITGA4, TGFβ1, α-SMA, and TIMP2 than the other groups, and there were significant reductions in the transcriptional levels of each factor after intervention with colchicine or SSAA, with a significant difference between the CCl 4 model group and the other groups (all P < 0.05). The CCl 4 model group had significantly higher protein expression levels of ITGA4, TGFβ1, α-SMA, TIMP2, and TIMP1 and a significantly lower protein expression level of MMP2 than the other groups, and intervention with colchicine or SSAA inhibited the expression of ITGA4, TGFβ1, α-SMA, TIMP2, and TIMP1 and promoted the expression of MMP2. Immunohistochemistry showed that the CCl 4 model group had significantly higher expression levels of TGFβ1 and α-SMA than the other groups, which was inhibited by intervention with colchicine or SSAA. The high-dose SSAA group had the most significant effect in reducing aminotransferases, improving lobular structure, and inhibiting the protein expression of liver fibrosis factors. Conclusion The high expression of ITGA4 in the liver is associated with the development of liver fibrosis, which is consistent with the increases in the expression of TGFβ1 and α-SMA. Inhibiting the expression of ITGA4 can provide more therapeutic targets for liver fibrosis and expand the anti-liver fibrosis mechanism of SSAA.

【Objective】 To investigate the relationship between hepatitis B virus X （HBx） protein and EGFR promoter， and the role of HBx protein in activating EGFR/PI3K/p-Akt signaling pathway and inhibiting apoptosis. 【Methods】 EGFR promoter plasmids were constructed and the relationship between HBx and EGFR promoters was characterized using a luciferase reporter assay. EGFR-overexpressing trophoblast cells were constructed， and EGFR expression in the overexpressing cells was knocked down using EGFR shRNA. The expression and localization of EGFR/PI3K/p-Akt were detected by Western blotting and confocal laser microscopy. Cell apoptosis was analyzed using flow cytometry. HBV plasmids carrying either full-length HBx or HBx with a deletion mutation （ΔHBx） and HBx plasmids were transfected into two types of trophoblast cells； HBx and PI3K/p-Akt protein expressions were detected by Western blotting. Cell apoptosis was analyzed using flow cytometry. 【Results】 Co-transfection of HBx and EGFR promoter plasmids in JEG-3 and HTR-8/Svneo cells significantly elevated the expression of EGFR promoter driven luciferase compared with the control group （P<0.01）. In EGFR-overexpressing cells， the expression of PI3K/p-Akt was significantly increased （P<0.01）， whereas the apoptosis rate was significantly decreased for JEG-3 cells and HTR-8/Svneo cells （both P<0.01）. These results were reversed in the EGFR-knock down group. When the intracellular HBx protein was expressed in JEG-3 and HTR-8 cells， PI3K/p-Akt protein expression was significantly increased （both P<0.05）， and the proportion of apoptosis was significantly decreased （both P<0.05）. 【Conclusion】 In placental trophoblast cells， HBx protein activates the expression of EGFR by acting on the EGFR promoter， and inhibits the apoptosis of trophoblast cells via the downstream EGFR/PI3K/p-Akt signaling pathway.

【Objective】 To investigate the effect and mechanism of inhibiting Yes-associated protein1 (YAP1) expression by verteporfin on proliferation, migration and invasion of human umbilical vein endothelial cells (HUVECs) exposed to hypoxia environment and the possible mechanisms that further affect placental angiogenesis in preeclampsia. 【Methods】 MTT method was used to detect the cell viability of HUVECs at different concentrations (0, 4, 8, 12 and 16 μg/mL) after 12 h and 24 h treatment with verteporfin under hypoxia and calculate the IC50 value to select the subsequent experimental drug concentration. Flow cytometry was made to analyze verteporfin’s effect on HUVEC apoptosis in hypoxic environment. The wound healing assay and Transwell invasion assay were used to determine the effect of verteporfin on HUVEC cell migration and invasion abilities under hypoxic environment. Angiogenesis test was used to detect the effect of verteporfin on the angiogenesis of HUVECs under hypoxic environment. The effects of verteporfin on the expression levels of YAP1 and TEAD1 in Hippo signaling pathway under normoxia and hypoxia were determined by Western blotting. 【Results】 Under hypoxic environment, verteporfin could inhibit the proliferation of HUVECs by calculating the IC50 value, the subsequent experimental group selected 16 μg/mL verteporfin to treat cells. Flow cytometry showed that verteporfin induced the apoptosis rate of HUVECs under hypoxia (P<0.01). The results of wound healing, Transwell invasion and the angiogenesis experiments confirmed that compared with the control group, verteporfin could inhibit the migration, invasion and angiogenesis of HUVECs in hypoxic environment (P<0.05). Western blotting assay indicated that under normoxia and hypoxia, the expressions of YAP1 and TEAD1 were reduced (P<0.01). 【Conclusion】 In hypoxic environment, verteporfin inhibits the proliferation of HUVECs by inhibiting the expressions of YAP1 and TEAD1, and reduces the migration, invasion and angiogenesis of HUVECs. It is confirmed that the Hippo-YAP1 signaling pathway may affect the placental angiogenesis of preeclampsia and participate in the occurrence of preeclampsia by regulating the proliferation and invasion of vascular endothelial cells.

Objective To explore the association between the GRIN3B gene and Tourette syndrome (TS) in children by screening mutations in the coding region of this gene.Methods Fifty-one children with TS and their parents in the Affiliated Hospital of Qingdao University from October 2015 to November 2016 were selected as an experimental group,41 cases of boys,and 10 cases of girls,aged 6-16 years[(9.78 ±3.64)years],while 60 people aged 22-45 years in the health examination center were selected in the control group,49 cases were male,1 1 cases were female,aged 22-45 years [(29.08 ± 2.89) years].DNA was extracted from 51 patients with TS,their parents and 60 controls.PCR was applied to amplify the encoding region of GRIN3B gene and Sanger sequencing was used to sequence,then GRIN3B sequencing results were compared with the NCBI gene encoding region sequence (NM_138690.2)to test whether these patients carried gene mutation and to verify the findings from their family.Results c.C460T gene variant of GRIN3B was found in 2 patients (p.P154S);c.T1187C (p.L396S) variant of GRIN3B gene was found in 10 patients and both of abnormal GRIN3B sites lead to changes in amino acid.The 2 peak sequencing maps were obtained by Sanger sequencing but nothing was found in their parents.Conclusion The mutation of GRIN3B gene may be related to the development of TS.

Objective To investigate the relationship between LUTS and degree,type of prostate histological inflammation in patients with benign prostatic hyperplasia (BPH).Methods From October 2013 to June 2016, 150 patients with BPH(62 -82 years old)were treated with selective TURP.After operation,the pathology examina-tion was practised.According to the international prostatitis histological classification standard,the patients were divid-ed into group A(without inflammation,18 cases),group B(inflammation around gland,45 cases),group C(gland inflammation,33 cases)and group D(mixed inflammation group,54 cases).There were no statistically significant differences among the 4 groups in age,course of disease,number of urinary retention and prostate volume(all P >0.05).The IPSS score before and 2 months after operation and the correlation between the IPSS and degree,type of histological inflammation were observed.Results In group A and C,the preoperative IPSS score and IPSS score at 2 months after surgery were (17.20 ±11.14)points,(8.45 ±5.14)points and (18.05 ±13.11)points,(9.85 ± 5.25)points respectively,there were no statistically significant differences between the two groups(t =0.485,1.294, all P >0.05).There were no statistically significant differences between group B[(22.55 ±11.85)points,(13.24 ± 6.46)points]and group D[(21.45 ±10.35)points,(12.36 ±6.04)points](t =0.824,1.436,all P >0.05).The group B had statistically significant differences compared with group A and group C (t =2.689,5.016,3.642,4.382, all P <0.05).The group D also had statistically significant differences compared with group A and group C (t =2.786,4.672,6.415,5.106,all P <0.05 ).There was positive correlation between the IPSS score before and 2 months after operation and the degree of inflammation in group B,D(B:r =0.698,0.795,D:r =0.546,0.485,all P <0.05),but there was no correlation in group C between them(r =0.182,0.170,all P >0.05 ).Conclusion There is a close correlation between LUTS and inflammation around gland in patients with BPH,and the LUTS can aggravate with the increased inflammation around gland and do not relate with the gland inflammation.

Objective To evaluate the clinical efficacy of bladder gasification and stasis dispersion in BPH patients with histological prostatitis after TUEP about IPSS /Qol score.Methods 126 BPH patients with histological prostatitis after TURP were divided into group A,B and C of equal number,and received oral placebo,tamsulosin,and herbal decoction a week after operation,respectively,for a course of 4 weeks.IPSS and Qol score were evaluated. Results After 5 weeks,121 cases finished the follow -up,the unfinished were 2 cases in group A,2 cases in group B and 1 cases in group C.The differences of the IPSS total scores,irritative subscore,obstructive subscore,quality of life score in group C(7.16 ±3.37)points,(4.22 ±2.70)points,(2.94 ±2.06)points,(1.47 ±1.44)points were statis-tically significant than those in group A[(13.77 ±4.50)points,(7.48 ±3.50)points,(6.29 ±2.63)points,(2.83 ± 1.72)points]and group B [(12.77 ±5.24)points,(8.05 ±3.88)points,(4.72 ±3.24)points,(3.07 ± 1.67)points](t =2.692,3.544,3.296,2.698 and t =2.326,3.532,2.418,2.414;all P <0.05).There was only obstructive subscore in group B better than that in group A(t =2.572,P <0.05).Conclusion Bladder gasification and stasis dispersion is effective for the treatment of BPH patients with histological prostatitis after TUEP about IPSS and Qol score.The clinical effect is indeed and satisfactory.

Objective:To investigate the status of oral hygiene behavior of the people of 1 8 yeas old and above in Guizhou prov-ince.Methods:A total of 9 280 adults were included by multi-staged sampling from 1 2 counties in Guizhou province of China.The double entry data were processed by Epiinfo 3.5.1 and statistically analysed by SPSS 1 9.0.Results:The survey of 9 280 subjects were permanent residents from 201 0 -201 1 .1 5.9% of the total subjects experienced dental visit,those from city and contrside were 23.0%,1 2.5%;male and female were 1 4.4%,1 7.1 %,respectively.5.6% of the them experienced tooth scaling,those from city and countryside were 9.9% and 3.4%,male and female were 4.9%,6.2% respectively.The subjects with 1 time and 2 time tooth brushing a day were 58.1 % and 28.6%,respectively.6% of the subjects did not brush teeth,mainly were those over 60 years (21 .3%),those of city residents was significantly lower than countryside(7.0% vs 4.0%).Conclusion:The oral heal behavior of Guizhou residents is poor,the education of oral health should be strengthened.

Objective:To analyze the levels of arachidonic acid (AA)and docosahexaenoic acid (DHA)in the breast milk of lactating mothers in Changchun City of Jilin Province, and to explore their influence factors. Methods:The lactating mother’s basic information was collected with questionnaire, and the breast milk of lactating mothers on postpartum 22-25 d was obtained and the 3-day 24-hour dietary recall method was used to investigate the dietary intake information of 514 healthy lactating mothers.The Food Composition Table of China 2009 was used to calculate the intakes of five kinds of polyunsaturated fatty acids (PUFAs) in diet of lactating mothers and the gold key nutrition expert system software for corresponding nutrient analysis was used to calculate the amount of various kinds of foods in the lactating mothers’daily diet.The levels of AA and DHA in breast milk were determined with gas chromatography and the linear regression was used to analyze the related factors of AA and DHA levels in breast milk.Results:①The average concentration of AA in breast milk of 514 lactating mothers was (0.08±0.04)g·100 g-1,and the average concentration of DHA was (0.05±0.04)g·100 g-1. ②The single factor correlation analysis results showed that the oil intake was both positively correlated with AA and DHA levels in milk of lactating mothers (r= 0.360,r=0.354,P<0.001),while the intakes of linoleic acid (LA), alpha linolenic acid (ALA),eicosapentaenoic acid (EPA),DHA,dairy and meats and seafood in diet were both negatively correlated with AA (r= -0.321,r=-0.280,r=-0.255,r=-0.299,r=-0.196 ,r=-0.306,P<0.05)and DHA (r=-0.315,r=-0.279,r=-0.175,r=-0.189,r=-0.248,r=-0.142,P<0.05).③The linear regression analysis results showed regression equation that dairy intake (β=-0.265)and EPA intake (β=-0.144)were both negatively correlated with the level of AA (P=0.009),and dairy intake was also negatively correlated with the level of DHA (β=-0.233,P<0.001).Conclusion:The AA and DHA levels in breast milk of lactating mothers didn’t increase with the increasing of intake of milk or dairy products in the study.Moreover there is a competitive relationship between n-6 and n-3 series polyunsaturated fatty acids in the process of metabolism.

OBJECTIVE: To explore the effect of ROCK inhibitor Y-27632 on the matrix metalloproteinase 2 and 9 (MMP2 and MMP9) gene expression and activity in tumor necrosis factor α (TNF-α)-treated human umbilical vein endothelial cell (HUVEC).
 METHODS: HHUVEC was divided into 3 groups, a control group, a TNF-α group, and a TNF-α plus Y-27632 group. The expressions of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), MMP2 and MMP9 were examined by real-time PCR. The MMP2/9 activity was measured by gelatin zymography.
 RESULTS: Compared to the control group, the mRNA expressions of ICAM-1, VCAM-1, MMP2 and MMP9 were increased TNF-α-treated cells, which were suppressed by ROCK inhibitor (P<0.01). The MMP2/9 activity was elevated in TNF-α-treated cells, which was reversed by ROCK inhibitor (P<0.05).
 CONCLUSION ROCK inhibitor can suppress TNF-α-induced inflammation in endothelial cells through down-regulation of MMP2/9.
Amides ; Cells, Cultured ; Down-Regulation ; Endothelial Cells ; Humans ; Intercellular Adhesion Molecule-1 ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinase 9 ; Pyridines ; Signal Transduction ; Tumor Necrosis Factor-alpha ; Umbilical Veins ; Vascular Cell Adhesion Molecule-1 ; rho-Associated Kinases