Objective To investigate the expressions and the relationship of FEZ1 and p16 in cervical intraepithelial neoplasia (CIN).Methods The expressions of FEZ1 and p16 in 93 cases of CIN and 10 cases of normal cervical specimens were detected by immunohistochemistry.Results The positive expression rates of FEZ 1 were 90.0％ (9/10) of normal cervical specimens,67.9％ (19/28) of CIN I,36.0％ (9/25) of CIN Ⅱ,22.5％(9/40) of CIN Ⅲ.The positive expression rates of p16 were 10.0％(1/10) of normal cervical specimens,32.1％(9/28) of CIN I,76.0％(19/25) of CIN Ⅱ,92.5％(37/40) of CIN Ⅲ.There were significant differences in the positive expression rates of FEZ1 and p16 between CIN Ⅰ and CIN Ⅲ (P ＜ 0.01),there was no significant difference in the positive expression rates of FEZ1 and p16 between CIN Ⅱ and CIN Ⅲ.The expression of FEZ1 and p16 was negatively correlation in CIN (r =-0.712,P＜ 0.01).Conclusion The abnormal high expression of p16 and abnormal low expression of FEZ1 in CIN may be involved in the occurrence and development of CIN,detecting the expressions of the two indexes may be helpful for clinical diagnosis.

BACKGROUND:Previous studies have demonstrated that transplantation of control ed release glial cellline-derived neurotrophic factor and bone marrow mesenchymal stem cells-derived neuron-like cells can effectively promote the motor function and sensory function recovery of rhesus monkeys with spinal cord injury.
OBJECTIVE:To validate whether co-transplantation of control ed release glial cellline-derived neurotrophic factor and bone marrow mesenchymal stem cells-derived neuron-like cells exhibits better protective effects on spinal cord glial scar of rhesus monkeys with spinal cord injury than celltransplantation alone.
METHODS:Twelve rhesus monkeys were col ected to prepare animal models of acute severe spinal cord injury using modified Al en’s method, and then randomly divided into three groups:experimental group, co-transplantation of control ed release glial cellline-derived neurotrophic factor and bone marrow mesenchymal stem cells-derived neuron-like cells;control group, simple transplantation of bone marrow mesenchymal stem cells-derived neuron-like cells;blank control group, PBS. After 5 months, paraffin specimens of the spinal cord were made for detection of morphological and compositional characteristics of glial scar, regeneration of nerve fibers in the scar, glial scar area, and average absorbance of glial fibril ary acidic protein.
RESULTS AND CONCLUSION:Glial scar in the injured spinal cord was composed of astrocytes and histocytes. Less spinal cord glial scar area and lower absorbance value could be observed in the experimental and control groups as compared with the blank control group (P<0.05). In addition, in the blank control group, neurofilament negative fibers could be observed in the glial scar, and astrocytes proliferated obviously. The experimental and control groups showed less fibers passed through the scar area. The glial scar area and average absorbance in the experimental group was lower than that in the control group (P<0.05). These findings suggest that compared with simple transplantation of bone marrow mesenchymal stem cells-derived neuron-like cells, co-transplantation of control ed release glial cellline-derived neurotrophic factor and bone marrow mesenchymal stem cells-derived neuron-like cells shows better protective effects spinal tissue structure after spinal cord injury, which may be one of mechanisms by which the number of glial scars is reduced to a greater extent.

AIM: To investigate the effects of metformin on airway inflammation, remodeling and neovascularization in a mouse model of chronic asthma and its possible mechanisms.METHODS: BALB/c mice were randomly divided into saline group, ovalbumin (OVA) group and OVA+metformin group, with 8 in each.At the end of OVA exposure, blood and bronchoalveolar lavage fluid (BALF) were collected for the measurement of OVA specific IgE and leukocyte counts.Lung tissue sections were stained with hematoxylin-eosin, periodic acid-Schiff and Masson's trichrome to detect inflammatory cell infiltration, goblet cell hyperplasia, and collagen deposition around the airway, respectively.Immunohistochemistry was used to evaluate the number and percentage area of new blood vessels (CD31+), and the protein level of phosphorylated AMP-activated protein kinase (p-AMPK) in the airway.RESULTS: Compared with saline group, the eosinophil percentage and OVA specific IgE in serum in OVA group were all increased obviously (P<0.01).Metformin inhibited the above increases (P<0.05).Compared with control group, a marked increase in inflammation infiltration, PAS+ cells and collage deposition in the airway mucosa in OVA group were observed.Metformin partially relieved the above changes.CD31+ vessels in the wall of bronchi showed the abundance of blood vessels observed in OVA group compared with control group, which was suppressed by the treatment with metformin (P<0.05).The protein level of p-AMPK was reduced in the lung tissue challenged with OVA as compared with control group (P<0.05), while metformin increased the protein level of p-AMPK (P<0.01).CONCLUSION: The protein level of p-AMPK in the airway in OVA group is attenuated.Metformin effectively inhibits airway inflammation, remodeling and neovascularization possibly via activating AMPK signaling pathway.

ObjectiveTo investigate the relationship between small intestinal bacterial overgrowth (SIBO) and the severity of hepatitis B cirrhosis. MethodsForty-seven patients with hepatitis B cirrhosis who visited Department of Gastroenterology, Zhengzhou People′s Hospital, from June 2014 to June 2015 were selected as cirrhosis group; these patients consisted of 11 Child-Pugh class A cases, 16 Child-Pugh class B cases, and 20 Child-Pugh class C cases. Fifteen healthy volunteers were selected as control group. The levels of procalcitonin, bilirubin, plasma albumin, and globulin were measured in all subjects. The lactulose hydrogen breath test (LHBT) was performed to measure small intestinal bacterial growth. Comparison of categorical data was made by chi-square test or Fisher's exact test. Comparison of normally distributed continuacs data between groups was performed by t test, The correlations of SIBO with procalcitonin, bilirubin, plasma albumin, and albumin/globulin ratio were investigated by Pearson correlation analysis. ResultsSIBO was found in 22 (46.8%) of the 47 patients with liver cirrhosis, consisting of 2 (18.1%) in the 11 Child-Pugh class A cases, 7 (43.7%) in the 16 Child-Pugh class B cases, and 13 (65.0%) in the 20 Child-Pugh class C cases. The prevalence of SIBO showed a significant difference between the controls and all patients with hepatitis B cirrhosis, Child-Pugh class B cases, or Child-Pugh class C cases (P=0.005, 0.037, or 0.001), but no significant difference between the controls and Child-Pugh class A cases (P=0556); there was a significant difference in the prevalence of SIBO between Child-Pugh class A cases and Child-Pugh class B/C cases (χ2=4727,P=0.030). The levels of procalcitonin, bilirubin, plasma albumin, and globulin were significantly correlated with LHBT set-value (r=0.895, 0.907, -0.810, and 0.755, respectively, all P＜0.001). ConclusionThe prevalence of SIBO increases with exacerbation of liver injury in patients with hepatitis B cirrhosis. The levels of procalcitonin, bilirubin, and globulin are positively correlated with SIBO, while plasma albumin level is negatively correlated with SIBO.

Glucocorticoids/urine* ; Humans ; Hydrocortisone ; Marathon Running ; Saliva/chemistry* ; alpha-Amylases/analysis*

Objective:To investigate the signaling pathway of inhibiting macrophage phagocytosis of TIR domain-containing protein encoded by Escherichia coli (TcpC) N-terminal ubiquitin ligase active fragments of uropathogenic Escherichia coli. Methods:Bioinformatics software was used to analyze the amino acid sequences and the function of TcpC N-terminal ubiquitin ligase active fragments as well as the functional sites. PCR was performed to amplify tcpc-330, tcpc-450 and tcpc-510 genes and a prokaryotic expression system was constructed to express the target proteins. The recombinant proteins rTcpC-N110, rTcpC-N150 and rTcpC-N170 were purified by Ni-NTA affinity chromatography. LPS in the recombinant proteins was removed by Detoxi-gel chromatography. The expression of MyD88 at protein and mRNA levels in macrophages incubated with rTcpC-N110, rTcpC-N150, rTcpC-N170 or rTcpC-TIR was detected by Western blot and qRT-PCR. The activation of NF-κB signal pathway and the levels of proinflammatory factors in macrophages incubated with the above TcpC protein fragments were measured by Western blot and ELISA, respectively. Results:Cys12, Trp104 and Trp106 in the N-terminal fragment of TcpC were crucial amino acids in maintaining its ubiquitin ligase activity. The target recombinant proteins rTcpC-N110, rTcpC-N150 and rTcpC-N170 were successfully expressed and purified. After Detoxi-gel chromatography, rTcpC-N110, rTcpC-N150 and rTcpC-N170 extracts were undetectable for LPS. TcpC ubiquitin ligase fragments inhibited the expression of MyD88 at protein level, but not affect its expression at mRNA level in macrophages. LPS-induced phosphorylation of NF-κB signaling pathway-related proteins p50 and p65 was significantly inhibited in macrophages treated with TcpC ubiquitin ligase fragments. Moreover, LPS-induced production of pro-inflammatory factors was also significantly inhibited.Conclusions:The recombinant proteins rTcpC-N110, rTcpC-N150 and rTcpC-N170 could inhibit the expression of MyD88 at protein level and suppress the activation of NF-κB signaling pathway, suggesting that they were closely related to the inhibition of innate immune activity of macrophages.

Objective:To investigate the role of TcpC in uropathogenic Escherichia coli (UPEC)-induced cystitis in mice and to preliminarily analyze the pathogenic mechanism. Methods:C57BL/6 mice were injected with 10 9 CFU wild-type UPEC CFT073 (CFT073 wt) or tcpc gene-deleted mutant (CFT073 Δ tcpc) from urethra into bladder to construct the mouse model of cystitis. The mice were sacrificed 3 d after infection and the bladders were taken to observe the gross pathological changes. Histopathological changes in bladder tissues were observed after HE staining. Immunohistochemistry was used to detect TcpC in bladder tissues. Bacterial loads in urine samples of UPEC-infected mice were counted by tenfold dilution method, and the presence of tcpc gene in the genomic DNA of bacteria from the bladder and urine samples of CFT073 wt-infected mice was measured by PCR. Real-time quantitative RT-PCR (qRT-PCR) and Western blot were performed to detect the expression of TcpC at mRNA and protein levels in macrophages after CFT073 wt infection. The influence of UPEC strains on the activation of NF-κB signaling pathway in macrophages were determined by Western blot. The levels of proinflammatory factors and the bacterial and cell activity after infecting macrophages with UPEC strains were detected by ELISA, laser confocal microscope and fluorescence microscope, respectively. Results:Compared with the mice with CFT073 Δ tcpc infection, CFT073 wt-infected mice had significantly enlarged bladder and severe neutrophil infiltration and abundant TcpC in bladder tissues. The number of bacteria in the urine of CFT073 wt-infected mice was significantly greater than that of the CFT073 Δ tcpc group. PCR results showed that the bacteria in bladder or urine were CFT073 wt. The expression of TcpC at both mRNA and protein levels in macrophages increased significantly after CFT073 wt infection. Moreover, in CFT073 wt-infected macrophages, the expression of IκBα was promoted and the phosphorylation of p65 and the production of proinflammatory factors were suppressed. TcpC was instrumental in the survival and invasion of CFT073 wt in macrophages. Conclusions:TcpC expression increased significantly in mice with CFT073 wt-induced cystitis. TcpC inhibited the activation of NF-κB signaling pathway and the production of proinflammatory factors in macrophages to improve the survival rate of CFT073 wt, which was closely related to the pathogenesis and immune evasion of UPEC.

Objective:To investigate whether the hypervirulent Klebsiella pneumoniae (hvKP) induces liver abscess through activating NLRP3 inflammasome. Methods:K1-hvKP and K35-non-hvKP bacterial suspensions were intraperitoneally injected into C57BL/6 mice to establish the models of liver abscess. Human peripheral blood neutrophils were sorted by immunomagnetic beads with CD45 + and Gr-1 + , and the purity was detected by flow cytometry. The concentrations of capsular polysaccharide of K1-hvKP and K35-non-hvKP were detected by total carbohydrate assay kit. The expression of IL-18 and IL-33 by neutrophils at mRNA and protein levels was detected by real-time fluorescence quantitative PCR and ELISA, respectively. The activation of NLRP3 inflammasome in neutrophils was detected by Western blot. Neutrophil extracellular trap formation (NETosis) was observed under confocal laser scanning microscope. Results:The C57BL/6 mice with K1-hvKP infection had significantly serious liver abscess as compared with the K35-non-hvKP-infected mice. The purity of human neutrophils was more than 95%. The concentration of capsular polysaccharide in K1-hvKP was significantly higher than that in K35-non-hvKP. Compared with K35-non-hvKP, K1-hvKP significantly promoted the neutrophils to express IL-18 and IL-33 at both mRNA and protein levels, enhanced the activation of NLRP3 and induced NETosis.Conclusions:This study suggested that hvKP could promote NETosis by activating NLRP3 inflammasome to cause liver abscess.

Objective: To investigate the effect of compound sophora flavescens injection on T helper cells 17 (Th17), regulatory T-cells (Treg cells) and the related cytokines in lung cancer patients with cirrhosis. Methods: A total of 63 patients with early non-small cell lung cancer and cirrhosis in Shouguang People's Hospital of Shandong Province from January 2016 to January 2018 were selected. All patients were scheduled to undergo surgical treatment. The patients were randomly divided into observation group (33 cases) and control group (30 cases) according to the random number table method. The patients in the control group were treated with liver protection, and the patients in the observation group were given compound sophora flavescens on the basis of the control group. After 3 months of treatment, the changes of liver fibrosis index, the levels of Th17 and Treg cells in peripheral blood and the changes of interleukin (IL)-10, IL-17, IL-6, and transforming growth factor (TGF)-β in serum were compared between the two groups. Results: After treatment, the alanine aminotransferase (ALT) of 15 cases (45.45%) in the observation group were normalized and 6 cases (20.00%) in the control group were normalized. There was a significant difference in ALT normalization rate between the two groups (P < 0.05). The levels of serum laminin (LN) and hyaluronic acid (HA) in the two groups after treatment were lower than those before treatment (all P < 0.05), and the serum LN [(156.74±30.52) ng/ml] and HA [(179.56±25.32) ng/ml] in the observation group after treatment was lower than those in the control group [(210.58±39.42) ng/ml and (203.75±28.79) ng/ml] (t values were 18.236 and 12.184, both P < 0.01). There was no significant difference in the level changes of type Ⅲ procollagen and type Ⅳ collagen between the two groups before and after treatment (all P > 0.05). After treatment, the levels of Th17 and Th17/Treg increased and the level of Treg cells decreased (all P < 0.05). After treatment, the levels of Th17 [(1.32±0.18)%] and Th17/Treg (0.23±0.04) in the observation group were higher than those in the control group [(1.21±0.17)% and 0.20±0.03] (t values were 3.201 and 1.087, both P < 0.01), while the level of Treg cells in the observation group was lower than that in the control group (P < 0.05). After treatment, the serum levels of IL-10, IL-17, IL-6, and TGF-β in the two groups were lower than those before treatment (all P < 0.05), and their levels in the observation group were lower than those in the control group (all P < 0.05). Conclusion Compound sophora flavescens injection can improve the liver function and fibrosis index of patients with lung cancer and cirrhosis, and it can regulate the levels of Th17, Treg cells and their related factors, which can be used as a clinical adjuvant.