Dental Implantation ; Humans ; Surgery, Computer-Assisted

Objective:To construct the human DcR3 expression vector and verify its expression in vitro. Methods: 915 bp human DcR3 gene CDS was amplified from porcine lung tissues,and was cloned into eukaryotic expression vector pEF1a-IRES-DsRed-Express2 which show red fluorescence. And then pEF1a-IRES-DsRed-Express2-DcR3 was transfected into LX-2 cells by FuGene HD. Expression of mRNA and protein lever of Human DcR3 were detected by RT-PCR and Western blot. Results:The levels of DcR3 gene transcription and translation in the hepatic stellate cells were significantly increased after transfection with pEF1a-IRES-DsRed-Ex-press2-DcR3 by RT-PCR and Western blot analysis. Conclusion: DcR3 expression vector was successfully constructed and highly expressed in LX-2 cells.

This study was designed to investigate the correlation between autophagy and polarization of macrophages in atherosclerosis (AS) plaque in arteriosclerosis obliterans amputees. Femoral artery specimens from arteriosclerosis obliterans amputees were performed hematoxylin and eosin (HE) staining, oil red O and immunofluorescence staining to observe the morphology of atherosclerotic plaque, phenotype of macrophages and autophagy in plaque; using real-time quantitative RT-PCR technology to detect the mRNA level of M1 and M2 type markers in arterial tissue; to analyze polarized signal pathway and autophagy protein levels in macrophages by Western blotting. Arterial specimens staining showed obvious lipid deposition and obvious infiltration of amount of foam cells and inflammatory cells. Macrophages were mainly expression M1 type in percentage in fibrous plaque. Although both M1 and M2 macrophages were upregulated in atheromatous plaque, the increase was dominant in M2 type in percentage. The level of autophagy was significantly higher in the atheromatous plaque than that of fibrous plaque. The expression of tumor necrosis factor- α (TNF-α), monocyte chemotactic protein-1 (MCP-1), inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6) and interleukin-12 (IL-12) mRNA was significantly higher in fibrous plaque than that of atheromatous plaque (P < 0.01 or 0.05), and arginase-1 (Arg-1), transforming growth factor-β (TGF-β), CD163 and interleukin-10 (IL-10) mRNA was significantly lower than that in atheromatous plaque (P < 0.01). The levels of p-STAT1 and NF-κB were significantly increased in fibrous plaque (P < 0.01), while p-STAT6 expression was significantly increased in atheromatous plaque (P < 0.01). The level of LC3-II was significantly higher in atheromatous plaque than that in fibrous plaque (P < 0.01). Macrophages in early atherosclerotic plaque were induced to M1 type through p-STAT1/NF-κB pathway and expressed moderate levels of autophagy; while macrophages in advanced plaques were induced to polarization of M2 type through p-STAT6 pathway. M2 macrophages expressed a higher level of autophagy than M1 macrophages.

Objective To investigate the effects of granulocyte macrophage colony-stimulating factor (GM-CSF) combined with interleukin-12 (IL-12) genes on apoptosis of hepatoma cells.Methods The hepatoma cell lines were cultured in vitro and were divided into four groups: GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group,negative control group (empty load group),respectively.The PIB-CMV3-GM-CSF and PIB-CMV3-IL-12 eukayotic expression vector was built,and 36 h after transfection,fluorescence microscope was used to detect the transfection effect;the expression level of IL-12,GM-CSF,p53,p38 and C-JUN mRNA were detected by RT-PCR,and Western blot was used to examine the expression level of IL-12,GM-CSF,p53,p38 and C-JUN protein.In addition,the flow cytometry was applied to detect cell apoptosis.Results Through fluorescence microscope,green fluorescence was observed in cells of GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group,indicating that the plasmid has successfully transferred into cells.In addition,the expression of p53mRNA in empty load group,GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group were 1.2±0.10,4.3±0.98,4.2±0.34,9.2±0.87,and the protein expression were 1.0±0.10,3.6±0.34,3.8±0.30,5.0±0.60.Compared with the empty load group,the expression level of p53 mRNA and protein were significantly increased in the three plasmid transfection groups (P<0.01).The expression of p53 mRNA and protein were significantly increased in co-transfection group than GM-CSF group and IL-12 group (P<0.01),while in the comparison with GM-CSF transfection group and IL-12 transfection group,the expression level of p53mRNA and protein in the co-transfection group could be improved to a higher degree(P<0.01).Meanwhile,p38 C-JUN mRNA expression levels in empty load group,GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group were as follows: 7.5± 0.9,3.5±0.45,3.7±0.25,1.0±0.11,while p38protein expression levels were 10.1±1.03,6.1± 0.67,7.1 ± 0.61,1.0 ± 0.12,respectively,C-JUN mRNA expression levels were 11.2 ± 1.20,4.1 ± 0.19,3.3 ± 0.30,1.0 ± 0.01,separately,C-JUN protein expression levels were 2.25 ± 0.2,1.8 ± 0.13,1.4 ± 0.12,1.0 ± 0.09.P38, C-JUN mRNA and protein levels were significantly reduced in the three plasmid transfection groups compared with the empty load group (P<0.01).The expression of p38,C-JUN mRNA and protein were reduced to a lower degree in co-transfection group than in GM-CSF transfection group and IL-12 transfection group (P<0.01).Flow cytometer showed that the hepatoma cell apoptosis rate of the empty load group,GM-CSF transfection group,IL-12 transfection group,co-transfection group were (3.43±0.9)%,(5.87±1.02)%,(7.32±1.1)%,(17.47±2.11)%,the rates of the three plasmid transfection groups were significantly higher than that of the empty load group (P<0.01).And the apoptosis rate was significantly increased in the co-transfected group compared with other plasmid groups (P<0.01). Conclusion The combination of GM-CSF and IL-12 could significantly accelerate the apoptosis of hepatoma cells by up-regulating the expression of p53,and down-regulating the expression of p38 and C-JUN.

ObjectiveTo investigate the effect of decoy receptor 3 (DcR3) gene on hepatocyte apoptosis, as well as the possible mechanism of action of DcR3 in this process. MethodsThe human liver cell lines were cultured in vitro, and pEF1α-DcR3 transfection group, pEF1α-IRES transfection group, and negative control group were established. The pEF1α-IRES-DsRed-Express2-DcR3 eukaryotic expression vector was constructed and transfected into human liver cell lines for 36 hours. qRT-PCR was used to measure the mRNA expression of DcR3, Fas ligand (FasL), α-smooth muscle actin (α-SMA), and transforming growth factor-β1 (TGF-β1), Western blot was used to measure the change in the protein expression of DcR3, and flow cytometry was used to measure apoptosis. An analysis of variance was used for comparison of continuous data between groups, and the least significant difference t-test was used for comparison between any two groups. ResultsAfter human liver cell lines were transfected with pEF1α-DcR3 for 36 hours, the pEF1α-DcR3 transfection group showed significant increases in the mRNA and protein expression of DcR3 compared with the pEF1α-IRES transfection group and negative control group (F=33 1695 and 14154, all P＜0.01). Compared with the other two groups, the pEF1α-DcR3 transfection group showed significant reductions in the mRNA expression of FasL, α-SMA, and TGF-β1 (F=269 4518, 20 7904, and 80678, all P＜0.01), which suggested that DcR3 inhibited the expression of FasL, α-SMA, and TGF-β1. Compared with the pEF1α-IRES transfection group and negative control group, the pEF1α-DcR3 transfection group showed a significant reduction in apoptosis rate (F=55863, all P＜0.01). ConclusionDcR3 can inhibit hepatocyte apoptosis and downregulate the mRNA expression of FasL, α-SMA, and TGF-β1.

Liver disease during pregnancy is substantially different from common liver diseases, and it is very important to raise the awareness of such disease and improve the diagnostic level. The literature on liver disease during pregnancy published in recent years is reviewed in this article, and the research advances in the pathogenesis, diagnosis, and treatment of liver disease during pregnancy are summarized.

Objective To explore the value of susceptibility-weighted imaging (SWI) for the assessment of chronic renal injury. Methods Thirty-nine patients with clinical diagnosis of chronic renal injury (RI group) who underwent routine renal MRI and SWI examination were retrospectively analyzed. They were divided into mild injured group (15 cases) and moderate to severe injured group (24 cases) by estimated glomerular filtration rate (eGFR). At the same time, 17 volunteers without chronic renal injury who had normal serum creatinine (Scr) and blood urea nitrogen were recruited as control group. All subjects underwent routine renal MRI and SWI examination. The ratios of cortex to medulla were measured and calculated in both kidneys' magnitude image and susceptibility weighted image, which were indicated as C/MMAG and C/MSWI. Independent sample t test was used to compare the differences of C/MMAG and C/MSWI between control group and RI group, and paired sample t test was used to compare the differences betweenC/MMAG and C/MSWI in each group. One-way ANOVA was used to compare the difference of C/MMAG and C/MSWI between the control group and the different RI groups. ROC was employed to assess the diagnostic efficacy of C/MMAG and C/MSWI in renal injury. Pearson linear correlation analysis was used to evaluate the correlation between C/MMAG, C/MSWI and eGFR, Scr in patients with renal injury. Results The C/MMAG and C/MSWI in the RI group were 1.101±0.039 and 1.071±0.046, respectively. C/MSWI was obviously lower than C/MMAG, and the difference was statistically significant (t=5.056, P<0.01). There was no significant difference between C/MMAG and C/MSWI in the control group (P>0.05). The C/MMAG and C/MSWI in the RI group were obviously lower than those in the control group, and the difference was statistically significant (t=4.564, 6.122;P<0.01).The C/MMAG and C/MSWI in the mild injured group and the moderate to severe injured group were significantly lower than those in the control group, the difference was statistically significant (P<0.05). While the differences of those between mild injured group and moderate to severe injured group showed no statistical significance (P>0.05). The area under ROC of C/MMAG and C/MSWI in diagnosis of renal injury were 0.853 and 0.952, respectively. C/MMAG was positively correlated with eGFR (r=0.460,P<0.01). Conclusions Susceptibility-weighted imaging can be used to assess chronic renal injury. Although it cannot reflect the degree of renal function damage, it has some value in the early diagnosis of mild renal injury.

Background: There is an urgent need to find reliable and rapid bovine tuberculosis (bTB) diagnostics in response to the rising prevalence of bTB worldwide. Toll-like receptor 2 (TLR2) recognizes components of bTB and initiates antigen-presenting cells to mediate humoral immunity. Evaluating the affinity of antigens with TLR2 can form the basis of a new method for the diagnosis of bTB based on humoral immunity. Objectives: To develop a reliable and rapid strategy to improve diagnostic tools for bTB. Methods: In this study, we expressed and purified the sixteen bTB-specific recombinant proteins in Escherichia coli. The two antigenic proteins, MPT70 and MPT83, which were most valuable for serological diagnosis of bTB were screened. Molecular docking technology was used to analyze the affinity of MPT70, MPT83, dominant epitope peptide of MPT70 (M1), and dominant epitope peptide MPT83 (M2) with TLR2, combined with the detection results of enzyme-linked immunosorbent assay to evaluate the molecular docking effect. Results: The results showed that interaction surface Cα-atom root mean square deviation of proteins (M1, M2, MPT70, MPT83)-TLR2 protein are less than 2.5 A, showing a high affinity.It is verified by clinical serum samples that MPT70, MPT83, MPT70-MPT83 showed good diagnostic potential for the detection of anti-bTB IgG and M1, M2 can replace the whole protein as the detection antigen. Conclusions Molecular docking to evaluate the affinity of bTB protein and TLR2 combined with ELISA provides new insights for the diagnosis of bTB.

Objective To investigate the occlusal force in the position of maximum intercuspation in the population with individual normal occlusion using a new occlusal analyzing equipment (TeeTester).Methods Twenty-nine volunteers aged from 21 to 32 years were recruited.Occlusal analysis was performed using TeeTester.An arch model of each subject was retrieved in order to measure the tooth width and to calculate the force applied on each tooth.Results The occlusal force in the position of maximum intercuspation was (900±361) N (range:335-1 727 N).The maximum occlusal force was correlated with the contacting area.The mean value of occlusal force on the molars ranged from 107 to 156 N,with the mean value on the first molar greater than that on the second molar within the same quadrant.The mean value of occlusal force ranged from 39 to 66 N on the premolars,and from 11 to 33 N on the front teeth.Conclusions There is a great variation of occlusal force in maximum intercuspation in individual normal occlusion.TeeTester occlusal analyzing system provides absolute occlusal force in kilogram and can be cowerted to N,which may assist clinical examination in patients.

Objective To summarize the clinical characteristics of Beh?et's disease (BD) in children with gastrointestinal involvement. Methods We retrospectively analyze the children BD with gastrointestinal involvement who were diagnosed in our hospital in recent 10 years. Results Twenty-two children were identified. The average age of onset was(6.1±4.0) years. The time from disease onset to clinical diagnosis was (1.2±2.1) years on average. Fifteen children had abdominal pain, diarrhea and hematochezia. Seven cases had positive endoscopic findings without any gastrointestinal symptoms. Twenty cases received corticosteroids therapy, 13 cases of them were treated with Cyclophosphamide/Methotrexate (CTX/MTX), 3 refractory cases were treated with biologics. Patients were followed up for (28±32) month on average. Eight patients' condition was stable, 7 patients were refractory, 3 patients died, 4 patients were lost to follow-up. At the same term, 5 patients without gastrointestinal involvement who received corticosteroids and CTX/MTX therapy were stable. Conclusion It is difficult to diagnose children BD at early stage. Gastrointestinal involvement may not be found, while the gastrointestinal endoscopy is of great importance in the diagnosis of the disease. Gluco-corticoid combined with immunosuppressive agents are effective. As to refractory patients, biological agent might be used although the recurrence is common. Compared with BD without gastrointestinal involvement, children BD with gastrointestinal involvement have serious condition and poor prognosis.