ObjectiveTo investigate the proliferation and apoptosis effects of adenovirus-mediated interleukin-24 (Ad-IL-24) gene on Karpas299 cells in vitro. MethodsThe Karpas299 cells were divided into blank control group, Ad-IL-24 group, and the adenovirus which carrying green fluorescent protein gene group (Ad-GFP group). Karpas299 cells of Ad-IL-24 group were infected by adding 200.0 μL Ad-IL-24, Karpas299 cells of Ad-GFP group were infected by adding 200.0 μL Ad-GFP, but Karpas299 cells of blank control group were treated by adding 200.0 μL PBS. Cells' proliferation inhibition rates of 3 groups were detected by cell counting kit (CCK-8) method at 12, 24, and 48 hours after treatment, respectively, and the cells' apoptosis rates of 3 groups were detected by flow cytometry at 48 hours after treatment. ResultsAd-IL-24 can suppress the growth of Karpas299 cells, and the inhibition rate increased over time. Compared with Ad-GFP group at the same time, the cell' proliferation inhibition rate of Ad-IL-24 group was higher at 12, 24, and 48 hours after treatment (P<0.05). In addition, the cells' apoptosis rate of Ad-IL-24 group was higher than those of Ad-GFP group and blank control group at 48 hours after treatment (P<0.05). ConclusionAd-IL-24 can suppress the growth of Karpas299 cells and induce the apoptosis of it.
Objective To examine the levels of interferon-gamma; (INF-gamma;), tumor necrosis factor-alpha; (TNF-alpha;) and interleukin-6(IL-6) in serum of patients with acute uveitis before and after treatment, and to explore the possible roles of those cytokines in the initiation and progression of the uveitis. Methods A series of 75 patients with acute uveitis,and 30 healthy persons from our hospital were investigated. The levels of INF-gamma;, TNF-alpha; and IL-6 in acute phase and convalescent phase were measured by the enzymelinked immunosorbent assay. Result The serum levels of INF-gamma;, TNF-alpha; and IL-6 in acute phase were significantly higher than that of the convalescent phase and the healthy controls (F=65.805/50.418/155.381, P=0.000). A significant negative correlation was found between the serum levels of INF-gamma;, TNF-alpha; and IL-6 in acute phase with their initial visual acuity(r=-0.656, -0.592 and -0.653, Plt;0.01). There was also a positive correlation among the serum levels of INF-gamma;, TNF-alpha; and IL-6(r=0.340, 0.467 and 0.338, Plt;0.05). Conclusions There are high serum levels of INF-gamma;, TNF-alpha; and IL-6 in patients with acute uveitis, and the cytokines levels were decreased after the treatment. The results suggested that the INF-gamma;, TNF-alpha; and IL-6 involved in initiation and progression of uveitis.
In perioperation period, the dynamic changes of solubla interleulcin-2 receptor (sIL-2R) in serum were determined by ELISA in 60 patients with gastric cancer (GC), and then was compared with those of 30 normal individuals and 40 selective patients who necieved common abdominal surgery. Results: At the day before and ten days after operation, the sIL-2R of patients with GC was higher than that of normal individual. But twenty days after operation, the sIL-2R reduced to as normal level. Conclusion: As a immunodepressive index, the sIL-2R of patients with GC was increased obviously, and after radical gastrectomy, it decreased gradually. So by determining sIL-2R, we can evaluate the immunologic function of patientswith GC.
Objective To observe the histopathologic features and expression patterns of tumor necrosis factor-alpha; （TNF-alpha;）, interleukin-1beta;（IL-1beta;） and Escherichia coli lipopolysaccharide (LPS) in the rat vitreous with LPS inducedendophthalmitis. Methods Wistar rats were randomly divided into saline control group (SC,136 rats）,endophthalmitis group （EO, 168 rats）and blank control group （BC,12 rats）.EO group received an intravitreal injection of 5 mu;l LPS; SC group received 5 mu;l sterile saline and no intervention for BC group.Six,12,24,48, and 72 hours,5 and 7 days after injection, intraocular inflammation were observed and the eyes and vitreous were collected for histopathological examination and measurement of TNF-alpha;, IL-1beta; and LPS expression. Results Severe inflammatory responses in the eyes were observed in EO group between six and 72 hours after LPS injection，ocular inflammation subsided seven days after LPS injection. In the vitreous, a peak neutrophil count was observed at 24 hours （1224.64plusmn;132.2） cells/eye that rapidly declined at 72 hours （342.25plusmn;47.7） cells/eye. The levels of TNF-alpha; and IL-1beta; in EO group were peaked at 24 hours with （996.18plusmn;89.45） and（5556plusmn;1440）pg/ L, respectively；Persisted at 48 hours and began to decline rapidly thereafter. Seven days after LPS injection, levels of TNF-alpha; and IL-1beta; returned to baseline with （22.16plusmn;5.84）and （73.7plusmn;18.7） pg/L, respectively. LPS concentration in EO group decrease rapidly at 72 hours with （11.03plusmn;3.41） ng and disappear on days 7 with (0.22plusmn;0.08) ng after LPS injection.Conclusions Massive neutrophils infiltration, high levels expression of TNF-alpha; and IL-1beta; and spontaneous elimination of bacterial elements in vitreous cavity were major pathologic characteristics in this experimental model. The expression patterns of TNF-alpha;,IL-1beta; were in accord with LPS clearance process.
ObjectiveTo investigate expressions of ALCAM/CD166, Bcl-2, and Ki-67 in breast infiltrative ductal cancer tissues, so as to assess the role of ALCAM/CD166 protein in the carcinogenesis and progression of breast infiltrative ductal cancer. MethodsThe expressions of ALCAM/CD166, Bcl-2, and Ki-67 proteins were examined by immunohistochemistry(ElivisionTM Plus) in 96 breast infiltrative ductal cancer specimens and 30 adjacent normal tissues of breast cancer specimens(control group). The relation between ALCAM/CD166 protein expression and patient's age, tumor diameter, histopathologic grade, axillary lymph node metastasis, or TNM stage of breast infiltrative ductal cancer was analyzed, and the correlation between ALCAM/CD166 expression and Bcl-2 or Ki-67 was analyzed too. Results①In 96 cases of breast infiltrative ductal cancer, the positive rate of ALCAM/CD166 protein expression was 79.2%(76/96), which was significantly higher than that in the control group〔10.0%(3/30), P < 0.01〕.②In the breast infiltrative ductal cancer tissues, the expression of ALCAM/CD166 was related to axillary lymph node metastasis(P < 0.05), but was not related to patient's age, tumor diameter, histopathologic grade, and TNM stage(P > 0.05).③The ALCAM/CD166 protein expression was positively related to Bcl-2(rs=0.307, P=0.001) and not related to Ki-67(rs=0.064, P=0.475). ConclusionALCAM/CD166 protein expression might be related to the apoptosis and metastasis of breast infiltrative ductal cancer cells and it could serve as an important marker for predicting biological behavior and prognosis of tumor.
Objective To examine the effects of carbon dioxide (CO2) pneumoperitoneum on local pancreas pathological changes, serum levels of amylase, IL-1, IL-6, and the positive rate of dissolubility adhesion molecule (CD11a/CD18 and CD11b/CD18) expression in rats with severe acute pancreatitis (SAP). Methods Fifty healthy male SpragueDawley (SD) rats were randomly divided into 3 groups: CO2 pneumoperitoneum group (n=20): SAP was induced by injecting 5% sodium taurocholate through retrogradely common biliopancreatic ducts via duodenal papilla， and then CO2 pneumoperitoneum was established at a pressure of 12 mm Hg (1 mm Hg=0.133 kPa) for 30 min; SAP group (n=20): The rats were treated as same as CO2 pneumoperitoneum group, except CO2 pneumoperitoneum； Simple operation group (n=10): Laparotomy was performed and nothing was done to duodenum and pancreas except for moving them softly. The blood samples were collected for examining serum levels of amylase, IL-1, IL-6, and the positive rates of CD11a/CD18 and CD11b/CD18 expression, and histopathologic examination of pancreas was performed. Results Compared with simple operation group, the pancreatic pathologic histology score, serum levels of amylase, IL-1, IL-6, and the positive rates of CD11a/CD18 and CD11b/CD18 expression were significantly higher in CO2 pneumoperitoneum group and SAP group (P=0.000). The levels of IL-1 and IL-6 were significantly lower in CO2 pneumoperitoneum group as compared to SAP group (P=0.000). There was no significant difference between CO2 pneumoperitoneum group and SAP group in pancreatic pathologic histology score (P=0.294), the level of serum amylase (P=0.073), the positive rates of CD11a/CD18 (P=0.155) and CD11b/CD18 expression (P=0.201). Conclusion CO2 pneumoperitoneum has inhibitory effect on the levels of IL-1 and IL-6, rather than the positive rates of CD11a/CD18 and CD11b/CD18 expression in SD rats with SAP.
Objective To investigate the expression and significance of Fork head /winged helix protein 3 (Foxp3) , retinoic acid-related orphan receptorγt (RORγt) , and interleukin-17 (IL-17) in Guinea pigs with emphysema. Methods Smoking and active immunization with elastin were separately used in guinea pigs to establish emphysema model. Then the destruction of lung tissue was assayed by measurement of the average radius of alveolar. The expressions of Foxp3 , RORγt, and IL-17 in lung tissue of the guinea pigs were detected by immunohistochemical technique. The results were compared with the normal control group by the analysis of variance or kruskal-Wallis test. Spearman rank correlation was used to analyze the correlation between the ratio of Foxp3/RORγt and IL-17, also the correlation between Foxp3/RORγt and the average radius of alveolar. Results In the smoking group and the active immunization group, the average radius of alveolar were significantly longer than the control group (Plt;0.05) . And the expression of Foxp3/RORγt was significantly unbalanced, with the number of Foxp3-positive cells decreased and RORγt-positive cells increased (Plt;0.05) . Meanwhile the level of IL-17 was significantly increased compared with the control group ( Plt;0.05) . The difference between the smoking group and the active immunization group was not significant (Pgt;0.05) . The ratio of Foxp3/RORγt was negatively correlated with the level of IL-17 and the average radius of alveolar. Conclusions Active immunization with elastin can induce emphysema in guinea pigs. The Foxp3/RORγt expression was unbalanced in lung tissue of guinea pigs with emphysema.This imbalance may be an important mechanism attributed to the disordered expression of CD4+ Treg cells and Th17 cells, which may be involved in autoimmune regulation and development of chronic obstructive pulmonary disease.
ObjectiveTo study the inhibitory effects of pigment epithelium derived factor (PEDF) on oxygen-induced retinal neovascularization in mice, and to investigate the possible involvement of interleukin-1β (IL-1β) in the neovascular-inhibitory function of PEDF. Methods A total of 140 postnatal day (P)7 C57BL/6 mice were randomly divided into normal control group, oxygen-induced retinopathy (OIR) model group, PEDF treatment group and PBS treatment control group. All mice except normal control group with their mothers were exposed to (75±2)% oxygen environment for 5 days and then kept in room air for another 5 days to establish the OIR model. Mice in normal control group were kept in room air only. At P12 and P14, respectively, mice in PEDF treatment group received intravitreous injections of 1 μl PEDF (2 μg/μl), while PBS treatment control group received the same volume of PBS (10 mmol/L, pH7.4).All mice were euthanized at P17 and eyes were isolated. The changes of retinal vessels were observed on retinal flat mounts and cryosections by fluorescence microscopy. Retinal specimens were prepared for IL-1β protein and mRNA analysis by Western blot and real time fluorescence quantitative reverse transcription-polymerase chain reaction (Real-time RT-PCR). ResultsChanges of retinal vessels had been viewed by fluorescence microscopy on flat-mounted retina, the relative retinal neovascularization areas were significantly increased in OIR model group compared with normal control group (t=15.02, P < 0.01), and the relative retinal neovascularization areas were obviously smaller in PEDF treatment group than those in PBS treatment control group (t=5.96, P < 0.01). Fluorescence staining revealed that retinal vascular tufts were extending from outer plexiform layer (OPL) to ganglion cell layer (GCL) of the retina along with multiple interconnections; Neovascular tufts in OIR model group and PBS treatment control group were presenting distinctly more than those of normal control group and PEDF treatment group. The specific expression levels of IL-1β protein in retinas of OIR mice by Western-blot analysis were higher than those of normal control group(t=3.35, P < 0.05), While these of PEDF treatment group showed a considerable decline in comparison with PBS treatment control group (P < 0.01), and there were no difference in normal control group and PEDF-treated group (F=11.764, P > 0.05). Similarly, expression levels of IL-1β mRNA tested by Real-time RT-PCR were obviously increased in the OIR model group when compared to normal control group(t=4.43, P < 0.01). After treated with PEDF, expression levels of IL-1β mRNA showed a considerable decrease when compared to PBS treatment control group (P < 0.01), and there were no difference in normal control group and PEDF-treated group (F=11.15, P > 0.05). ConclusionsPEDF can inhibit oxygen-induced retinal neovascularization. The mechanism may be related to that PEDF can downregulate the expression of IL-1β in retina.