ObjectiveTo investigate the role and mechanism of P-selectin glycoprotein ligand-1 (PSGL-1) in hydrochloric acid-induced acute lung injury (ALI) in mice.MethodsWild-type mice (WT) and PSGL-1 knockout mice (PSGL-1 -/-) were randomly subjected to normal saline (NS) or hydrochloric acid (HCl) challenged group. The mice were intratracheally instilled with NS or HCl (1 μl/g weight) into the left lung with a catheter. After 2 hours, respiratory function index enhanced pause (Penh), PaO2 and PaO2 were analyzed. The wet to dry weight ratio (W/D) of the left lung and total protein concentration in bronchoalveolar lavage fluid (BALF) were measured. The number of leukocytes in BALF was counted too. Targeted lung tissue was processed for further HE or immunohistochemistry staining. Meanwhile, the expressions of interleukin-6 (IL-6), IL-1β, nuclear factor-κB (NF-κB), IκBa and p-IκBa in lung tissue were measured.ResultsThe Penh (4.77±1.22 vs. 5.80±0.84) and PaCO2 [(63.7±3.9) mm Hg vs. (74.4±7.4) mm Hg] in the PSGL-1 knockout mice were significantly lower than those in the WT mice after HCl stimulation (P<0.05), while the PaO2 was higher than that in the WT mice [(81.0±7.1) mm Hg vs. (62.0±8.9) mm Hg, P<0.05)]. The lung W/D ratio (4.86±0.15 vs. 5.22±0.20), protein concentration [(3.71±0.64) μg/μl vs. (4.74±0.98) μg/μl] and total leukocyte count [(13.00±2.18) ×107/L vs. (49.42±3.35) ×107/L] in BALF were significantly lower in the PSGL-1 knockout mice challenged with HCl than those in the WT mice (P<0.05). Besides, the protein expressions of IL-6, IL-1β, p65 and p-IκBa in the PSGL-1 knockout mice were lower than those in the WT mice after HCl instillation, while the IκBa expression was higher than that in the WT mice (P<0.05). More numbers of neutrophils and macrophages were found in the lung of the WT mice than the PSGL-1 knockout mice challenged with HCl. However, the differences of above values between the WT mice and the PSGL-1 knockout mice instilled with NS were not found, all of which were significantly lower than the correspongding HCl group except for IκBa (P<0.05).ConclusionPSGL-1 may play important roles in the development of HCl-induced ALI via the NF-κB signaling pathway and inflammation.
Objective To explore the role of nuclear factor kappa B(NF-KB)in the pathogenesis of chronic obstructive pulmonary disease(COPD)and the therapeutic efects of glucocorticoid．Methods Twenty-four Wistar rats were randomly divided into three groups，ie．normal control，COPD model and prednisone preventive treatment group．Rat COPD model Was established by exposing the rats to cigarette smoke daily．Prednisone Was given through stomachal injection on altemate days．After COPD model Was set up，bronchoalveolar lavage(BAL)Was performed．Total cell counts and neutrophil counts in BALF were examined．Pathological changes of lung tissue Was observe0 by hematoxylin-eosin staining．The morphological indices of pulmonary emphysema(MLI，MAN and PAA)Was measured by a computerizedimage analyzer and compared in three groups．NF-KB expression in lung tissues were detected by immunohistochemistry assay．Rults Emphysema Was confirmed by three morphological indices in COPD model group compared to those of normal control group[MLI：(97.97±11.10)×10-6m vs (47．23±2．80)×10-6 m，MAN：(95.98±l4.89)×106 ／m vs (164.21±9．30)×106 ／m ，PAA：(64 ±5.7)％vs (44±2.7)％，Plt;0.01]．Total cell counts and neutrophil counts in BALF of COPD model group were significantly higher than those of control group[（5.76±0.29）×108/L vs （1.64±0.12）×108/L，（1.26±0.25）×108/L vs （0.099±0.065）×108/L，Plt;0.01]．After the preventive treatment with prednisone，MLI，MAN and PAA were significantly changed[（57.66±4.62）×10－6mvs （97.97±11.10）×10－6m，（111.40±16.92）×106個/m2 vs （95.98±14.89）×106個/m2，Plt;0.01；（58±6.1）% vs （64±5.7）%，Plt;0.05]，which indicated that airway inflammation and emphysematous injury in preventive treatm ent group were milder than those of COPD mode1．Total ceil counts and neutrophil countsin BALF were found in preventive treatment group as compared to those of COPD model[[（3.18±0.29）×108/L vs （5.76±0.29）×108/L，（0.57±0.12）×108/L vs （1.26±0.25）×108/L，Plt;0.01].The percentage of positive cells of NF-KB nuclear staining in bronchiolar epithelial ceils was significantly increased in the COPD group than that in the control group[（29.02±1.25）% vs （12.17±1.13）%，Plt;0.01]，but was significantly decreased in the preventive treatment group[(19.23±1.18)％vs (29.02±1.25)％，Plt;0．01]．Conclusions NF-KB may be responsible for the persistence and amplification of inflammation in COPD through neutrophil recruitment and activation．Prednisone may suppress airwayinflammation in COPD by inhibiting NF-KB．
Objective The usefulness of measurement of nuclear DNA content elevation for diagnosis of early hepatocellular carcinoma was evaluated by a study of 186 patients with liver cirrhosis. Methods Nuclear DNA content was measured using an automatic image analysis system.Results ①Hepatocellular carcinoma was found in 37 patients during 10 years follow-up, the cumulative incidence of hepatocellular carcinoma was 19.89%. ②The incidence of hepatocellular carcinoma increased with the increase of the patterns of α-fetoprotein (AFP), 5c exceeding rate (5cER), FORM PE, but positive predictive value of 5cER was the highest of three parameters, the difference among all groups was significant by the χ2 test (Ｐ＜0.05). ③When 5cER joined AFP for monitoring development of hepatocellular carcinoma, the incidence of hepatocellular carcinoma was 72.00%, which was significantly higher than that of 5cER or AFP alone, the difference between groups was highly significant (Ｐ＜0.01). Conclusion Patients who had 5cER levels of 3%-5% or more, who had transient increases in 5cER or who had both, should be treated as being in a super-highrisk group for hepatocellular carcinoma. Frequent and careful examination by ultrasonography of such patients is recommended. It is important that measurement of 5cER join with AFP in cirrhotic patients monitored for early development of hepatocellular carcinoma.
ObjectiveTo investigate the molecular mechanism of osteoclast differentiation induced by Staphylococcal peptidoglycan (PGN-sa). MethodsRaw264.7 cells were stimulated with PGN-sa and with PGN-sa+SC75741[a potent inhibitor of nuclear factor κB (NF-κB) activation] in a concentration of 200 ng/mL. The protein expression of nuclear factor of activated T-cells cytoplasmic 1 (NFATc1) was tested at 0, 1, 2, and 3 days; the proteins related to osteoclast differentiation of extracellular regulated protein kinases (ERK), p38, c-Jun N-terminal kinase (JNK), NF-κB, inhibitor of NF-κB (IκB-α), Akt, and the phosphorylation forms of p38, ERK, JNK, Akt, NF-κB were measured at 0, 5, 10, 20, 40, and 60 minutes by Western blot. In addition, Raw264.7 cells were stimulated with PGN-sa in the concentrations of 100 ng/mL (group A), 200 ng/mL (group B), 400 ng/mL (group C), and with PBS (group D) for 1, 2, and 3 days; the expression levels of tumor necrosis factor α (TNF-α), interleukin 1α (IL-1α), and IL-6 were detected by ELISA. ResultsThe results of Western blot showed that the expression of NFATc1 increased gradually with time, showing significant difference between different time points (P<0.05). However, after SC75741 was added, the expression of NFATc1 was inhibited at 2 and 3 days, showing significant difference when compared with no addition of SC75741 (P<0.001). After stimulation of PGN-sa, the expression of IkB-α decreased significantly at 5 and 10 minutes when compared with those at the other time points (P<0.001), and returned to normal at 20 minutes. Meanwhile, the expression of p-NF-κB increased significantly at 5 and 10 minutes when compared with those at the other time points (P<0.001), and returned to normal at 20 minutes; and the expression of p-NF-κB at 5 minutes was significantly higher than that at 10 minutes (P<0.001). After the addition of SC75741, there was no change in the expressions of IκB-α and p-NF-κB, showing no significant difference between different time points P>0.05). Moreover, the expressions of ERK, p38, JNK, NF-κB, Akt, p-p38, p-ERK, p-JNK, and p-Akt showed no significant change between different time points P>0.05). ELISA results showed that there were no expressions of TNF-α and IL-1α in groups A-D at different time points. The expression of IL-6 had an increasing trend with time prolonged in each group, showing significant differences between different time points (P<0.05). Moreover, at 1 day after culture, the expression of IL-6 showed no significant difference among groups P>0.05). At 2 and 3 days after culture, the expression of IL-6 in groups A-C showed an increasing trend and was significantly higher than that in group D, showing significant difference among groups (P<0.05). ConclusionPGN-sa can promote osteoclast differentiation through NF-κB signaling pathway, and IL-6 may play a role in this process.
Objective Series of compl icated molecule signal pathway are involved in the bone regeneration. To explore the possibil ity of nuclear factore kappa B (NF-κB) which is taken as the “key activation” during the fracture healing and provide the laboratory evidence for the gene therapy of nonunion or delayed union of fractures. Methods Thirtythree adult male Wistar rats (weighing 180-220 g) were selected and divided randomly into 4 groups: group A (the control group, n=3), the rigth lower segments of radius were injected with normal sal ine 0.3 mL for 7 days, once per day; group B (Bay 11-7082 injection group, n=6), the middle and distal radius were injected with normal sal ine containing 50 μmol/L NF- κB inhibitor Bay 11-7082 0.3 mL for 7 days, once per day; group C (fracture group, n=12), the right middle and distal radius were cut by a sharp scissors to form per fracture model; and group D (Bay 11-7082 treatment group, n=12), based on group C, 0.3 mL of 50 μmol/L Bay 11-7082 were injected into the fracture site for 7 days, once per day. The callus tissues were harvested at 3, 7, 14, and 28 days after fracture for Western blot analysis, alkal ine phosphatase (ALP) activity assessment, prostaglandins E2 (PGE2) production assay, and histological observation. Results The rats of all groups were survivaltill the experiment completion. At 3 and 7 days after injection, there was no significant difference in the ALP activity and PGE2 production between group B and group A (P gt; 0.05); but group C was significantly higher than group A (P lt; 0.01) and group D was significantly lower than group A (P lt; 0.01). The expressions of NF-κB p65, bone morphogenetic protein 7 (BMP-7), and inhibitor of DNA binding 2 (Id2) were observed at fracture sites of 4 groups. There was no significant difference in the expressions of NF-κB p65, BMP-7, and Id2 between group B and group A (P gt; 0.05); the expressions of NF-κB p65 and BMP-7 were significantly higher and the expression of Id2 was significantly lower in group C than group A (P lt; 0.01); and the expressions of NF-κB p65 and BMP-7 were significantly lower and the expression of Id2 was significantly higher in group D than group A (P lt; 0.01). The histological observation showed that a lot of osseous callus formed in group C at 14 and 28 days, but osseous callus just began to form in group D at 28 days. Conclusion NF-κB p65 can facil itate early fracture heal ing of rat radius by elevating the PGE2 production and regulating BMP-7 and Id2 expression.
Objective To study the relationship between smoking as well as smoke cessation and lung inflammation of chronic obstructive pulmonary disease( COPD) , and its possible mechanism. Methods Thirty-two Wistar rats were randomly divided into a control group, a low-dose smoking group, a high-dose smoking group, and a smoke cessation group. The expressions of NF-κB and E-selectin in rats lung were detected by immunohistochemistry and hybridization in situ. Results The mRNA and protein expressions of NF-κB in bronchial endothelial cells of the high-dose smoking group( 0. 43 ±0. 01, 0. 41 ±0. 04) , the lowdose smoking group( 0. 41 ±0. 01, 0. 40 ±0. 01) , and the smoke cessation group( 0. 39 ±0. 01, 0. 37 ±0. 03) , were significantly higher than the control group( 0. 29 ±0. 06, 0. 28 ±0. 06) ( all P lt; 0. 05) , with the high-dose smoking group and the smoke cessation group significantly higher than the low-dose smoking group ( both P lt;0. 05) . The mRNA and protein expressions of E-selectin had the similar pattern with NF-κB. The expressions of E-selectin mRNA and protein were positively correlated with the expression of NF-κB mRNA and protein respectively ( r = 0. 80, r = 0. 89, P lt; 0. 01) . Conclusions Smoking can result in high expressions of NF-κB and E-selectin in a dose dependent manner. Smoke cessation can relieve airway inflammation and is an effective measure for preventing COPD.
OBJECTIVE:To investigate the diagnostic meaning of MRI in intraocular tumors. METHODS:Forty-six cases of confirmed intraocular tumors,including choroidal melanoma(20 cases),retinoblastoma(18 cases),Coats disease(6 cases)and choroidal hemangioma(2 cases),were studied with MRI and compared with ultrasonography and CT. RESULTS:In making discoveries about intraocular tumors,there were no sighificant difference between MRI and B-ultrasonography or CT （P＞0.03，chi;2=1.0716）while there were highly statistic sighificance in dediding characters and position （P＜0.01，deceding character chi;2=29.8314，positionchi;2=13.659）of them. CONCLUSION:Among the examinations to find out about the position,character and secondary pathological insults of in traocular tumors MRI might be more available than CT and ultrasonography. （Chin J Ocul Fundus Dis,1997,13:93-95 ）
ObjectiveTo detect the metabolites of the serum and joint fluid from rabbits’ osteoarthritis model with 1H nuclear magnetic resonance spectroscopy (NMR) technique, study the metabolic differences and connections of serum, synovial and cartilage of rabbits after the articular cavity injection of sodium hyaluronate, and explore osteoarthritis and metabolic mechanism in the process of treating sodium hyaluronate using sodium hyaluronate, thus provide new ideas and basis of the specific mechanisms in the treatment of osteoarthritis via sodium hyaluronate.MethodsWe selected 30 healthy New Zealand white rabbits, 6 months old, and randomly divided them into three groups as follows: blank control group, model phosphate buffer saline (PBS) liquid injection group and model injection of sodium hyaluronate group, with 10 rabbits in each group. Ten weeks after surgery, all experimental animals were put to death and observed in correlation studies regarding general condition, imaging examination, and histological examination. Metabolites 1H NMR detection and data preprocessing were performed in the serum and joint fluid samples.ResultsThe results considering general condition, general sample observation, imaging examination and histology indicated advantages in sodium hyaluronate group over PBS group. Metabolomics analysis showed statistically significant changes of metabolites in the serum and joint fluid compared with the PBS group and the blank control group (P<0.05). According to the relevant ways of differences metabolites retrieval, analysis found that the effect of sodium hyaluronate on osteoarthritis might be related to protein biosynthesis, amino acid circulation, the metabolic process of pyruvic acid, gluconeogenesis and other metabolic pathways.ConclusionsBased on the research of 1H-NMR metabolomics, the results suggest that the effect of sodium hyaluronate on osteoarthritis is mainly related with the activation of protein metabolism, abnormal lipid and energy metabolic pathways. This study provides new ideas and basis on the concrete mechanism in the treatment of knee osteoarthritis using sodium hyaluronate.
ObjectiveTo determine the nuclear factor kappa B (NFkB) activity in peripheral blood mononuclear cells (PBMC) in patients with acute cholangitis of severe type (ACST) and correlate the degree of NFkB activation with severity of biliary tract infection and clinical outcome.MethodsTwenty patients with ACST were divided into survivor group (14 cases) and nonsurvivor group (6 cases). Other 10 patients undergoing elective gastrectomy or inguinal hernia repair were selected as control group. Peripheral blood samples were taken 24 hours after operation, PBMC was separated and nuclear proteins were isolated from PBMC, and NFkB was determined with electrophoretic mobility shift assay (EMSA). The levels of TNFα, IL6 and IL10 in plasma were determined by using an enzymelinked immunoassay (ELISA). ResultsThe NFkB activity was 5.02±1.03, 2.98±0.51 and 1.02±0.34 respectively in three groups. It was increased in all patients with ACST, versus the control group (P＜0.05), and the patients of nonsurvivor group had higher levels of NFkB activation than those of survivor group (P＜0.05). The levels of TNFα and IL6 were (496.28±52.35) ng/L and (578.13±67.72) ng/L in nonsurvivor group; (284.47±39.41) ng/L and (318.67±34.92) ng/L in survivor group; (89.43±10.39) ng/L and (101.27±13.47) ng/L in control group. All patients with ACST had increased levels of TNFα and IL6, which were many fold greater than that of control group, and there was an evidence of significantly higher levels in nonsurvivor group than in survivor group (P＜0.05). All patients had also increased levels of IL10 as compared to control group (P＜0.05), but the IL10 concentrations in plasma were not significantly higher in nonsurvivors than that of in those survivors (Pgt;0.05). ConclusionNFkB activation in PBMCs in patients with ACST
Dysplasia of gastric stump mucosa in 47 cases was studies.Nuclear DNA contents were measured with an automatic imagie analysis system.The results showed that the mean values of the nuclear DNA contents,area,perimeter,maximum diameter,minimum diameter increased with the increase of severity of dysplasia in gastric stump mucosa(Plt;0.01);where as nuclear form factor decreased with the increase of severity of dysplasia in gastric stump mucosa(Plt;0.05).Severe dysplasia is similar to that of gastric stump cancer in the DNA ploidy histogram.Our results indicate biological behaviour of gastric stump mucosa dysplasia.This study suggests that DNA contents analysis may be used as an important reference for grading,screening,and treating dysplasia of gastric stump mucosa.