ObjectiveTo observe effect of interferon-γ in preventing intestinal adhesion following abdominal surgery in rats.MethodsA total of 60 Wistar rats were selected, which were randomly divided into a sham operation (SO) group, model group, dexamethasone (DXMS) group, and interferon-γ group, then the interferon-γ group was randomly divided into a low, medium, and high concentrations subgroups. Except for the SO group, the laparotomies with file friction caecum were performed for all the other groups to establish the intestinal adhesion model. SO group and model group were intraperitoneally injected with saline for 4 mL/kg. The 10 mg/kg dexamethasone was injected into the abdominal cavity of rats in the DXMS group. The 7.5×104, 1.5×105, and 3.0×105 U/kg interferon-γ concentrations were injected into the abdominal cavity of rats in the low, medium, and high concentrations subgroups respectively. The Nair grading of cecum adhesion degree was assessed on the 8th day after the surgery, then the histopathological change was observed by the HE staining under the microscopy and the hydroxyproline content in the cecum tissue was detected.Results① The intestinal adhesion: Compared with the SO group, the intestinal adhesions occurred in all the other groups and the degrees of intestinal adhesions evaluated by the Nair grading were more significantly serious (P<0.05), which in the DXMS group and the medium and high concentrations of interferon-γ subgroups were significantly reduced (P<0.05) as compared with the model group, which in the high concentration of interferon-γ subgroup was significantly reduced (P<0.05) as compared with the DXMS group. ② The microscopic observation of histopathological results: Compared with the DXMS group, the high concentration of interferon-γ could effectively reduce the occurrence of fibrous tissue and inflammatory cell infiltration, the intestinal wall muscular layer structure was complete, a few inflammatory cells scattered in the infiltration. ③ The hydroxyproline content: The contents of hydroxyproline in the cecum tissue of the model group, DXMS group, and interferon-γ subgroups were significantly increased (P<0.05) as compared with the SO group, which of the DXMS group and medium and high concentrations of interferon-γ subgroups were significantly decreased (P<0.05) as compared with the model group, which of the high concentration of interferon-γ subgroup was significantly decreased (P<0.05) as compared with the DXMS group.ConclusionInterferon-γ has a preventive and therapeutic effect on postoperative intestinal adhesion and cecum injuries.
Objective To explore the effect of NaOH on the surface morphology of three-dimensional (3D) printed poly-L-lactic acid (PLLA) mesh scaffolds. Methods The 3D printed PLLA mesh scaffolds were prepared by fused deposition molding technology, then the scaffold surfaces were etched with the NaOH solution. The concentrations of NaOH solution were 0.01, 0.1, 0.5, 1.0, and 3.0 mol/L, and the treatment time was 1, 3, 6, 9, and 12 hours, respectively. There were a total of 25 concentration and time combinations. After treatment, the microstructure, energy spectrum, roughness, hydrophilicity, compressive strength, as well as cell adhesion and proliferation of the scaffolds were observed. The untreated scaffolds were used as a normal control. Results 3D printed PLLA mesh scaffolds were successfully prepared by using fused deposition molding technology. After NaOH etching treatment, a rough or micro porous structure was constructed on the surface of the scaffold, and with the increase of NaOH concentration and treatment time, the size and density of the pores increased. The characterization of the scaffolds by energy dispersive spectroscopy showed that the crystal contains two elements, Na and O. The surface roughness of NaOH treated scaffolds significantly increased (P<0.05) and the contact angle significantly decreased (P<0.05) compared to untreated scaffolds. There was no significant difference in compressive strength between the untreated scaffolds and treated scaffolds under conditions of 0.1 mol/L/12 h and 1.0 mol/L/3 h (P>0.05), while the compression strength of the other treated scaffolds were significantly lower than that of the untreated scaffolds (P<0.05). After co-culturing the cells with the scaffold, NaOH treatment resulted in an increase in the number of cells on the surface of the scaffold and the spreading area of individual cells, and more synapses extending from adherent cells. Conclusion NaOH treatment is beneficial for increasing the surface hydrophilicity and cell adhesion of 3D printed PLLA mesh scaffolds.
Objective To study the relationship between expression of nm23, CD44 in gastric carcinoma and lymph-node metastasis and prognosis. Methods Expression of nm 23, CD44H and CD44V6 in 105 cases of gastric carcinoma were assayed by immunohistochemistry. Among them, 59 cases were followed up. Results The incidences of nm23, CD44H and CD44V6 protein positivity in gastric carcinoma were 44.8%, 54.3% and 48.6% respectively. The positive expression of nm23, CD44V6 protein in human gastric carcinoma tissues was related to the differentiation, depth of invasion, TNM stage and prognosis (Ｐ＜0.05), but expression of CD44H was not correlated with other clinicopathologic indices. The reactivity to these three antibodies were correlate with metastasis of lymph nodes (Ｐ＜0.01 for CD44V6 and Ｐ＜0.05 for nm23, CD44H). Conclusion Expression of the standard form of CD44 (CD44H) might be useful in observing the progression of the disease, wile CD44V6 and nm23 hold promise as a prognostic indicator.
【Abstract】 Objective To explore the preventing effects of TGF-β1 antibody (TGF-β1Ab) compounded with fibringlue (FG) on postoperative adhesions of flexor tendon. Methods Seventy-two Leghorn chickens were randomly divided into 4 groups (groups A, B, C and D), 18 chickens for each group, and the long flexor tendons of the 3rd and 4th toes in zone Ⅱ of all chickens were transversed and sutured with the 4-strand cruciate repair technique to make defect models. In group A, 0.2 mL TGF-β1 Ab was appl ied at repair site. In group B, 0.2 mL FG was appl ied at repair site. In group C, 0.2 mL TGF-β1Ab and FG was appl ied at repair site. In group D, 0.2 mL normal sodium was appl ied at repair site. At 1, 3 and 8 weeks after operation, the tendons of 6 chickens in each group were harvested for morphological and histological evaluation. Six specimens of each group were obtained for biomechanical test at 3 and 8 weeks. Results The gross observation showed that the differences ingrading of tendon adhesion were not significant among 4 groups at 1 week after operation (P gt; 0.05), but the differences were significant between groups A, B, D and group C at 3 and 8 weeks after operation (P lt; 0.05). Histological observation showed that collagen fibers arranged irregularly in groups A, B and D, but arranged regularly in group C at 3 and 8 weeks after operation. At 3 weeks after operation the gl iding excursion ratio of the tendon in groups A, B, C and D were 0.45 ± 0.05, 0.40 ± 0.10, 0.79 ± 0.09 and 0.25 ± 0.07 respectively ; the simulated active flexion ratio were 0.61 ± 0.02, 0.67 ± 0.03, 0.91 ± 0.03 and 0.53 ± 0.04 respectively; the work of flexion were(18.00 ± 0.77), (17.80 ± 1.13), (27.60 ± 1.73) and (15.60 ± 1.27)?/N respectively. There were significant differences between group C and other three groups (P lt; 0.05). The tendon anastomosis breaking strengthwere (14.2 ± 1.9), (15.2 ± 2.2), (16.0 ± 2.2) and (14.7 ± 2.7) N, showing no significant differences among 4 groups (P gt; 0.05).At 8 weeks after operation, the gl iding excursion ratio of the tendon in groups A, B, C and D were 0.45 ± 0.07, 0.43 ± 0.08, 0.80 ± 0.09 and 0.29 ± 0.05 respectively; the simulated active flexion ratio were 0.61 ± 0.02, 0.63 ± 0.03, 0.92 ± 0.03 and 0.53 ± 0.03 respectively, the work of flexion were (18.30 ± 0.84), (18.60 ± 0.80), (27.90 ± 1.24) and (15.30 ± 0.75) ?/N respectively. There were significant differences between group C and other three groups (P lt; 0.05). The tendon anastomosis breaking strength were(51.9 ± 3.0), (51.4 ± 1.4), (53.3 ± 1.3) and (52.3 ± 2.2) N, showing no significant differences among 4 groups (P gt; 0.05). Conclusion TGF- β1Ab compounded with FG could significantly prohibit the formation of fibrous adhesions without interfering with the heal ing process.
ObjectiveTo observe the effects of p21 activated kinase 4 (PAK4) on the mitochondrial function and biological behavior in retinal vascular endothelial cells. MethodsThe experimental study was divided into two parts: in vivo animal experiment and in vitro cell experiment. In vivo animal experiments: 12 healthy C57BL/6J male mice were randomly divided into normal control group and diabetes group, with 6 mice in each group. Diabetes mice were induced by streptozotocin to establish diabetes model. Eight weeks after modeling, quantitative real-time polymerase chain reaction and Western blots were performed to detect the expression of PAK4 in diabetic retinas. In vitro cell experiments: the human retinal microvascular endothelial cells (hRMEC) were divided into three groups: conventional cultured cells group (N group), empty vector transfected (Vector group); pcDNA-PAK4 eukaryotic expression plasmid transfected group (PAK4 group). WB and qPCR were used to detect transfection efficiency, while scratching assay, cell scratch test was used to detect cell migration in hRMEC of each group. In vitro white blood cell adhesion experiment combined with 4 ', 6-diamino-2-phenylindole staining was used to detect the number of white blood cells adhering to hRMEC in each group. The Seahorse XFe96 cell energy metabolism analyzer measures intracellular mitochondrial basal respiration, adenosine triphosphate (ATP) production, maximum respiration, and reserve respiration capacity. The t-test was used for comparison between the two groups. Single factor analysis of variance was used for comparison among the three groups. ResultsIn vivo animal experiments: compared with normal control group, the relative expression levels of PAK4 mRNA and protein in retina of diabetic mice were significantly increased, with statistical significance (t=25.372, 22.419, 25.372; P＜0.05). In vitro cell experiment: compared with the N group and Vector group, the PAK4 protein, mRNA relative expression and cell mobility in the hRMEC of PAK4 group were significantly increased, with statistical significance (F=36.821, 38.692, 29.421; P＜0.05). Flow cytometry showed that the adhesion number of leukocytes on hRMEC in PAK4 group was significantly increased, and the difference was statistically significant (F=39.649, P＜0.01). Mitochondrial pressure measurement results showed that the capacity of mitochondrial basic respiration, ATP production, maximum respiration and reserve respiration in hRMEC in PAK4 group was significantly decreased, with statistical significance (F=27.472, 22.315, 31.147, 27.472; P＜0.05). ConclusionOver-expression of PAK4 impairs mitochondrial function and significantly promotes leukocyte adhesion and migration in retinal vascular endothelial cells.
【Abstract】Objective To study the correlation between focal adhesion kinase (FAK) expression and biological behavior of invasion and metastasis in gastric carcinoma. MethodsThe immunoreactivity of FAK was revealed by immunohistochemical method in gastric carcinoma tissues,canceradjacent tissues, normal gastric mucosa,and 200 regional lymph nodes in 50 collected specimens when radical resection of gastric carcinoma were carried out. ResultsThe percentage of FAK bly positive immunoreactivity were 10.0%（5/50）， 20.0%（10/50） and 78.0%（39/50） in normal gastric mucosa， cancer-adjacent tissues and gastric carcinoma tissues respectively. The b immunoreactivity was obviously higher in gastric carcinoma tissues than that in normal gastric mucosa or canceradjacent tissues (P＜0.01). There were no difference between normal gastric mucosa and canceradjacent tissues (Pgt;0.05). The percentage of bly positive immunoreactivity of FAK were 68.8%（22/32） and 33.3%（6/18） in cases with metastasis and without metastasis of lymph node respectively. The immunoreactivity of FAK in cases with metastasis of lymph node was significantly higher than that without metastasis of lymph node (P＜0.05).The b immunoreactivity of FAK were 87.2%（136/156） and 40.9%(18/44) in withmetastatic lymph node and withoutmetastatic lymph node. The immunoreactivity of FAK in withmetastatic lymph node was significantly higher than that withoutmetastatic lymph node （P＜0.01）. The percentage of FAK bly positive immunoreactivity showed correlation with the cellular differentiation and depth of infiltration of gastric carcinoma. The deeper infiltration and lower differentiation, the ber expression rate was obtained (P＜0.05), which showed no correlation with Borrmann type, location and size of tumor (Pgt;0.05). ConclusionIncreased immunoreactivity of FAK is an important role of invasion and metastasis for gastric carcinoma cells. Detection of FAK expression in cancer tissues can be helpful to understand the carcinogenic biological behavior of gastric carcinoma as well as to make judgment and treatment of prognosis of patients.
ObjectiveTo investigate the effect of Smad4 on the fibrosis of tendon derived fibroblasts (TDFs) induced by transforming growth factor β1(TGF-β1) by targeted regulation of miRNA219-5P (miR219-5P). MethodsThe tendons donated by the volunteers were harvested to isolate and culture TDFs. The 3rd generation cells were used for experiment. Chemically synthesized miR219-5P mimics, miR219-5P inhibitor, and negative control sequences were transfected into TDFs. The gene expression of miR219-5P in TDFs was detected by real-time PCR, and the protein expression of Smad4 in TDFs was detected by Western blot at 48 hours after transfection. The combining sites of miR219-5P and Smad4 in 3'UTR district were predicted by informatics software. Wild type and mutant type reporter gene expression vectors were constructed and then targeted verification was carried out by the luciferase reporter gene test. Transfected TDFs were then induced by TGF-β1. The proliferation activity of the cells were measured by the cell counting kit 8 after culturing for 24, 48, and 72 hours. The expressions of fibrosis related proteins in TDFs were detected by Western blot at 72 hours. ResultsAfter TDFs were transfected by miR219-5P mimics, miR219-5P expression was significantly up-regulated, but the expressions of Smad4 was decreased subsequently (P<0.05). Intracellular expression of miR219-5P was inhibited by miR219-5P mimics inhibitor, however, the protein expression of Smad4 was significantly increased (P<0.05). Luciferase reporter gene test showed that luciferase activities were significantly decreased in pGL3-WT-Smad4+mimics group, but were significantly increased in pGL3-WT-Smad4+inhibitor group when compared with pGL3-WT-Smad4 transfected group (P<0.05), but no significant difference was found between GL3-MT-Smad4+mimics and pGL3-MT-Smad4+inhibitor groups (P>0.05). Cell proliferation and the fibrosis related proteins were increased in TGF-β1 induced TDFs, however, decreased in TGF-β1 induced TDFs after transfected by miR219-5P inhibitor (P<0.01). ConclusionmiR219-5P can significantly inhibit fibrosis of TDFs induced by TGF-β1 by down-regulating Smad4 expression.
ObjectiveTo systematicly evaluate expression of epithelial cell adhesion molecule (EpCAM) in colorectal cancer (CRC) and its correlation with clinicopathologic characteristics of patient with CRC.MethodsPubMed, Web of Science, Cochrane Library, Embase, CNKI, Wanfang, VIP, and other databases were searched comprehensively. The retrieved literatures were imported into Endnote X9. The data about the expression of EpCAM in the CRC and the relationship between EpCAM expression and clinicopathologic characteristics of patients with CRC were screened and extracted. RevMan 5.3 software was used for meta-analysis.ResultsA total of 5 396 patients with CRC were included. The meta-analysis results showed that the expression rate of EpCAM in the CRC tissues or blood was significantly higher than that in the benign colorectal tumor and normal tissue or blood (P<0.05). The high expression rates of EpCAM in the Dukes C+D stage, tumor diameter >3 cm, infiltration state of tumor margin, with lymph node and distant metastasis of the CRC were significantly higher than those in the A+B stage, tumor diameter ≤3 cm, dilated state of tumor margin, without lymph node and distant metastasis (P<0.05).ConclusionResults of this meta-analysis suggest that expression of EpCAM might be related to some clinicopathologic characteristics (carcinogenesis, Dukes stage, tumor size, tumor margin morphology, lymph node metastasis, distant metastasis) of patients with CRC.
ObjectiveTo investigate the effects and clinical significance of edaravone on serum tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and intercellular adhesion molecule-1 (ICAM-1) in elderly patients with obstructive sleep apnea hypopnea syndrome (OSAHS).MethodsA total of 90 elderly patients with moderate to severe OSAHS confirmed by polysomnography were recruited from North China University of Science and Technology Affiliated Hospital in February 2016 to October 2017. According to random number table method the OSAHS patients were randomly divided into group A (n=30), group B (n=30) and group C (n=30). Group A received continuous positive airway pressure treatment for six months, group B received edaravone therapy and continuous positive airway pressure treatment for six months, and group C only received edaravone therapy for six months. The changes of serum TNF-α, IL-6 and ICAM-1 were detected by enzyme-linked immunosorbent assay before and after treatment.ResultsThe differences of serum TNF-α, IL-6 and ICAM-1 before treatment in the three groups were not statistically significant (P>0.05). Compared with before treatment, the levels of serum TNF-α, IL-6 and ICAM-1 decreased in the three groups (P<0.05). After six months of treatment, the levels of serum TNF-α, IL-6 and ICAM-1 decreased in group A and group B compared with group C (P<0.05), and decreased significantly in group B compared with group A (P<0.05).ConclusionEdaravone can inhibit the expressions of serum TNF-α, IL-6 and ICAM-1 in elderly patients with moderate to severe OSAHS, and thereby reduce vascular endothelial dysfunction and injury.
ObjectiveTo explore the feasibility and safety of the artificial pneumoperitoneum and gastrointestinal contrast CT imaging, and imaging diagnostic value on abdominal wall adhesion to intestine after operation. MethodsThirtynine patients with adhesive intestinal obstruction after operation relieved by conservative therapy were included from January 2008 to November 2009. After the artificial pneumoperitoneum established by injection of gas into abdominal cavity and gastrointestinal comparison by oral administration low concentration of meglucamine diatrizoate, CT scan imaging was performed and the radiographic results were compared with surgical findings. ResultsFour patients refused surgery and discharged, so enterolysis was performed in the remaining patients. The surgical findings were consistent with radiographic results. It was showed by laparoscopic operation that intestinal obstruction caused by the fibrous adhesions and the intestine did not adhere to the abdominal wall in eight patients with fibrous adhesion diagnosed by CT. Of eighteen patients with the abdominal wall septally adhered to the intestinal, the surgical findings showed the intestine and the abdominal wall formed “M”type adhesions and omentum adhesions in sixteen patients underwent open operation, and clear fat space was showed in eight patients and close adhesion was found in another eight patients between the intestine and abdominal wall. Of thirteen patients with the abdominal wall tentiformly adhered to the intestinal, the surgical findings showed the intestine and the abdominal wall formed continuous and tentiform adhesions and omentum adhesions to the intestine in eleven patients. After the followup of 6-18 months (mean 9 months), incomplete intestinal obstruction occurred in one patient and was relieved by conservative treatment. One patient with discontinuous discomfort in abdomen after operation did not receive any treatment. The other patients were cured. ConclusionThe artificial pneumoperitoneum and gastrointestinal contrast CT imaging can accurately show the location, area, and structure composition of the postoperative abdominal wall adhesion to intestine, which is safety, simple, and bly repeatable, and a better imaging method for the diagnosing of abdominal wall adhesion to intestine after operation.