1.Effects of Clostridium butyricum combined with glutamine on the expression of intestinal tight junction protein in stress mouse model
Donghui CHEN ; Yong YU ; Zhiqiang LIU ; Fuguang LI ; Pengyuan ZHENG
Chinese Journal of Digestion 2014;34(8):544-549
Objective To investigate the mechanism of the permeability of intestinal mucosa in the pathogenesis of irritable bowel syndrome (IBS) and the interventional effects of Clostridium butyricum combined with glutamine.Methods According to random number method,fifty BALB/c mice were divided into control group,experimental control group,glutamine group,Clostridium butyricum group and combination group.IBS mice model was established by water-avoidance stress (WAS) experiment.The defecating time of mice and fecal water content were detected by dyed stool after mice gavaged with methylcellulose (1.5%).The pathological injury of intestine was assessed by hematoxylin and eosin staining.The visceral sensitivity was evaluated by colorectal distention test (CRD).The changes of the permeability of intestine was evaluated by detecting the changes of serum D-lactic acid (D-LA),level of diamine oxidase (DAO),expressions of intestinal epithelial cells (IEC) cell tight junction protein (TJ) (occludin-1,claudin-1,zonula occludens-1 (ZOL-1)) at protein level.The interventional effects of Clostridium butyricum combined with glutamine were evaluated.t test was performed for comparison between groups,and analysis of variance was used for comparison among multi-groups.Results Compared with the control group,the defecating time of experimental control group was significantly shorten ((100.40±14.80) min vs (75.88±12.20) min and water content of fecal significantly increased ((54.76±9.98)% vs (74.95±7.15)%,t =3.692 and 4.023; P=0.002 and 0.002).The lowest threshold of visceral sensitivity significantly decreased ((40.87 ± 4.82) mmHg (1 mmHg=0.133 kPa) vs (27.80±3.18) mmHg; t=8.761,P<0.01),while the mucosal pathological injury score significantly increased (0.50±0.15 vs2.60±0.97; t=6.034,P<0.01).The level of D-LA ((1 476±246.8) ng/L vs (913.6±90.1) ng/L)) and DAO ((3 391.0±256.9) vs (5 096.0±725.2) ng/L) significantly increased (t=40.920 and 29.810; both P<0.05),and the expression of tight junction protein ZOL-1 (0.165±0.005 vs0.119±0.003),occludin-1 (0.104±0.016 vs 0.022±0.006) significantly decreased (t=19.830 and 19.830; both P<0.01).Compared with the experimental control group,after intragastric intervention the defecating time of glutamine group,Clostridium butyricum group and combination group increased ((90.50±3.78),(97.56±8.79) and (99.89±11.90) min and water content of fecal decreased ((69.33±6.71)%,(58.07±8.97)% and (56.74±8.12)%) and the differences were statistically significant (F=10.020 and 8.740; both P<0.01).The results of Clostridium butyricum group and combination group were good (F=2.481 and 4.874; both P<0.05).And the lowest threshold of visceral sensitivity significantly increased ((31.80±2.69),(36.04±5.06) and (38.93±3.30) mmHg; F=2.420,P<0.05),the result of combination group was the best (F=3.550,P<0.01).Jejunal mucosal injury was significantly reduced (2.00 ± 0.94,1.30 ± 0.68 and 1.30±0.48; F=11.350,P<0.01).After intragastric intervention,serum levels of D-LA ((1 370.0± 78.9),(1 066.0±155.5) and (1 039.0±129.0) ng/L) and DAO ((4 808.0±477.4),(3 713.0± 595.0) and (3 725.0±615.9) ng/L) of glutamine group,Clostridium butyricum group and combination group significantly decreased (F=37.480 and 27.670; both P<0.01).The level of ZOL-1(0.126± 0.014,0.125±0.006,0.138±0.004) and occludin 1 (0.037±0.013,0.073±0.028,0.078±0.027) of glutamine group,Clostridium butyricum group and combination group significantly increased,and the differences were statistically significant (F=5.867 and 10.630; both P<0.05).The change of ZOL-1 of combination group was more than that of Clostridium butyricum group (t =5.457,P < 0.05).Conclusions WAS experiment can induce visceral hypersensitivity,increase the permeability of intestine and reduce the function of intestinal epithelial barrier.Clostridium butyricum and glutamine are effective in the recovery of visceral hypersensitivity and the permeability of mucosal epithelia cells.
2.Effects of Bifidobacterium infantis on intestinal corticotropin releasing factor receptors and mast cells activation in a murine model of irritable bowel syndrome
Simeng LIU ; Pengyuan ZHENG ; Fuguang LI ; Donghui CHEN
Chinese Journal of Microbiology and Immunology 2014;(8):571-576
Objective To investigate the effects of Bifidobacterium infatn ison the expression of in -testinal corticotropin releasing factor ( CRF) receptors and how the peripheral CRF receptors activate mast cells in a murine model of irritable bowel syndrome (IBS).Methods Thirty BALB/c male mice were ran-domly divided into three groups including control group , model group and Bifidobacterium infantis group. The mouse model of IBS was established by using chronic restraint stress .Mice in Bifidobacterium infantis group received daily intragastrical administration of Bifidobacterium infantis for 14 days.Mice in control and model groups were treated with equal volume of saline .Then all mice were killed after the assessment of weight and abdominal withdrawal reflex ( AWR) .The levels of histamine , tryptase and tumor necrosis fac-tor-α( TNF-α) in serum samples were detected by ELISA .The expression of CRF in colonic mucosa was analyzed by immunohistochemistry .The expression of CRF-R1 and CRF-R2 in mast cells and the number of mast cells in colonic mucosa were detected by double immunofluorescence staining assay .The expression of CRF-R1 and CRF-R2 at mRNA level in colon were detected by reverse transcription polymerase chain reac-tion ( RT-PCR) .Results Compared with control group , the levels of histamine , tryptase and TNF-αin pe-ripheral blood samples , the expression of CRF-R1 and CRF-R2 at mRNA level , and the number of mast cells, CRF-R1+mast cells and CRF-R2+mast cells in colonic mucosa were increased significantly in model group (P<0.05), but were remarkably down-regulated with the treatment ofB ifidobacterium infantis (P<0.05).Conclusion Bifidobacterium infantis could reduce the activation of mast cells in a murine model of IBS by inhibiting the expression of CRF-R1 and CRF-R2 in intestinal mast cells .
3.Effects of aryl hydrocarbon receptor on Th17 cells activation in a murine model of irritable bowel syndrome
Yuying WANG ; Donghui CHEN ; Pengyuan ZHENG ; Fuguang LI
Chinese Journal of Microbiology and Immunology 2014;(4):299-303
Objective To detect the dynamic Th17 cells in a murine model of irritable bowel syn-drome ( IBS) and to study the effect of aryl hydrocarbon receptor ( Ahr) on Th17 cells activation .Methods Thirty BALB/c male mice were randomly divided into three groups including experiment group ,control group and Ahr antagonist group .A murine model of IBS was established by perfusing three nitrobenzene sulfonic acid (TNBS) into the colon of mice.Equal volume of saline was used to set up the control .The mice in Ahr antagonist group were intraperitoneally injected with 10 μg Ahr antagonist for four consecutive days .All mice were evaluated for visceral hypersensitivity and colonic mucosal inflammation .Mesenteric lymph nodes (MLNs) and peripheral blood mononuclear cells (PBMCs) were detected by flow cytometry through staining Th17 cells.The distribution of Ahr and IL-17A in colon and the number of Th17 cells activated by Ahr (Ahr and IL-17A double positive ) were detected by double immunofluorescence staining .Results ( 1 ) The percentage of Th17 cells in MLNs was significantly increased in experiment group followed by those in Ahr antagonist group and control group (P<0.05).(2)Compared with control group,the number of Th17 cells in peripheral blood samples was significantly increased in experiment group and Ahr antagonist group ( both P<0.05 ) ,but there was no difference between Ahr antagonist group and experiment group ( P=0.642 ) .( 3 ) The number of Ahr-activated Th17 cells ( Ahr+IL-17A+) was significantly increased in experiment group (10.00±1.58) as in comparison with that in control group (3.80±0.83,P<0.05),but the number was de-creased with Ahr antagonist intervention ( 5.80 ±0.83 , P<0.05 ) .Conclusion The number of activated Th17 cells was increased in MLNs and peripheral blood samples from mice with IBS .Ahr played an important role in the activation of Th17 cells in intestines.However,the number of Ahr-activated Th17 cells in intestinal mucosa and the proportion of Th 17 cells in MLNs could be down-regulated through blocking Ahr .
4.A pilot study on Lauren classification of advanced gastric carcinoma using double contrast-enhanced ultrasonography
Shiyan LI ; Pintong HUANG ; Yanping LI ; Liang WANG ; Fuguang HUANG ; Zhiqiang ZHENG ; Zongmin WANG
Chinese Journal of Ultrasonography 2008;17(12):1041-1044
Objective To investigate the clinical value of double contrast-enhanced ultrasonography (DCUS)in diagnosing Lauren classification of advanced gastric carcinoma.Methods Fifty-eight patients with gastric cancer proved by endoscopic biopsy underwent preoperative DCUS examination,and the findings were compared with postoperative pathologic results via hematoxylin-eosin and alcian blue-periodic acid schiff(AB-PAS)staining.Results In 58 patients,34 cases were diagnosed as intestinal type and 24 cases as diffuse type by pathology after operation.In intestinal type,30 cases were enhanced as global homogeneous intensity by using DCUS.another 2 cases as"lamellar"model and the other 2 cases as heterogeneous intensity.However,in diffuse type,22 cases were enhanced heterogeneously with"barrier"signs in 5 cases,"map"signs in 9 cases and"lamellar"signs in 8 cases.the remaining 2 cases of diffuse type enhanced as homogeneous model.The sensitivity and specificity of heterogeneous enhancement in diagnosing diffuse type of advanced gastric cancer was 91.7%and 88.2%respectively,and the Youden's index was 0.8.Conclusions DCUS could be considered as a new valuable method to distinguish the Lauren classification and evaluate the invasiveness,metastasis and prognosis in patients with gastric carcinoma.
5.The value of double contrast-enhanced ultrasonography in Borrmann classification of advanced gastric carcinoma
Liang WANG ; Pintong HUANG ; Yaping ZHAO ; Fuguang HUANG ; Yanping LI ; Shiyan LI ; Zhiqiang ZHENG ; Zongmin WANG
Chinese Journal of Ultrasonography 2008;17(11):965-968
Objective To investigate the clinical value of double contrast-enhanced ultrasonography (DCUS)in Borrmann classification of advanced gastric carcinoma preoperatively. Methods A total of 69 patients with advanced gastric carcinoma were examined by ultrasound after taking oral contrast agent and bolus injection of SonoVue preoperatively.A comparison with final pathologic results was made after surgery. Results The accuracy of oral contrast-enhanced ultrasonography and DCUS in Borrmann classification of advanced gastric cancer were 78.3%and 91.3%,respectively.There was a significant difference between two methods(X2=4.087,P<0.05).In addition,Kappa value of two methods were 0.73 and 0.90,respectively.Conclusions DCUS is a new valuable method for Borrmann classification with its high accuracy preoperatively.
6.Pilot study of double contrast ultrasonography for differentiation degree of gastric cancer
Yanping LI ; Pintong HUANG ; Yan YANG ; Shiyan LI ; Liang WANG ; Zhilin ZHAO ; Fuguang HUANG ; Zhiqiang ZHENG
Chinese Journal of Ultrasonography 2008;17(10):865-868
Objective To investigate the value of oral and intravenous contrast enhanced ultrasonography-double contrast ultrasonography (DCUS) in the evaluation of differentiation degree of gastric cancer. Methods Fifty-three patients who had been diagnosed as gastric cancer by gastroscope and confirmed by pathology after operation were examined by DCUS, and they were divided into moderately to well differentiated group and poorly differentiated group. The correlation of size and enhancement features of the lesions was compared with differentiation degree of gastric cancer. Results The size and enhancement distribution of lesions were markedly correlated with the degree of differentiation. The baseline intensity(BI)in poorly differentiated group was lower than that in well and moderately differentiated group[(0.52±1.12)dB vs (1.53±1.87) dB, P<0.05], but enhanced intensity(EI) was higher than that in the second group [(26.22±3.69)dB vs (23.69±5.02)dB, P<0.05]. Conclusions DCUS could initial evaluate the degree of differentiation of gastric cancer before operation.
7.IFN-γ inhibits acute allergic airway inflammation in mice
Qiuhong GAN ; Lili JIANG ; Qianyang LI ; Liu YANG ; Zhaojing ZHANG ; Fuguang LI
Chinese Journal of Microbiology and Immunology 2017;37(8):573-579
Objective To investigate the inhibitory effect of IFN-γ on acute allergic airway inflammation induced by IL-33 in mice.Methods Twenty-four female C57BL/6 mice (6-8 weeks) were randomly divided into four groups: IL-33 model group, IFN-γ treatment group, IL-33+IFN-γ treatment group and PBS control group.A mouse model of acute allergic airway inflammation was induced by IL-33.Samples of bronchial alveolar lavage fluid (BALF) and lung tissues were collected.Group 2 innate lymphoid cells (ILC2s) and eosinophils were analyzed by flow cytometry.Levels of IL-5 and IL-13 in the supernatants of lung homogenate and BALF were measured by ELISA.Expression of IL-5, IL-13 and ST2 at mRNA level was detected by real-time PCR.Pathological changes in lung tissues were observed following hematoxylin and eosin (HE) and periodic acid-Schiff (PAS) staining.Results Compared with the PBS control group, no infiltration with inflammatory cells, goblet cell hyperplasia or mucus secretion was observed in the IFN-γ group;the numbers of ILC2s and eosinophils were not affected by IFN-γ;the levels of IL-5 and IL-13 in the supernatants of BALF and lung homogenate, and the expression of IL-5, IL-13 and ST2 at mRNA level in lung tissues were not significantly changed by IFN-γ (P>0.05).Compared with the PBS control group, massive infiltration with inflammatory cells, excessive mucus secretion, increased numbers of ILC2s and eosinophils, up-regulated levels of IL-5 and IL-13 in the supernatants of BALF and lung homogenate, and enhanced expression of IL-5, IL-13 and ST2 at mRNA level in lung tissues were detected in the IL-33 model group (P<0.05).Compared with the IL-33 model group, the combined treatment with IL-33 and IFN-γ significantly alleviated inflammatory cell infiltration, inhibited mucus secretion, reduced the numbers of ILC2s and eosinophils, down-regulated the levels of IL-5 and IL-13 in the supernatants of BALF and lung homogenate, and suppressed the expression of IL-5, IL-13 and ST2 at mRNA in lung tissues (P<0.05).Conclusion IFN-γ can inhibit the proliferation of eosinophils and ILC2s induced by IL-33, and reduce the secretion of IL-5 and IL-13, which indicates that IFN-γ has an inhibitory effect on acute allergic airway inflammation induced by IL-33 in mice.
8.Application of echocardiography in the diagnosis of neonatal ductus arteriosus aneurysm
Hongju KOU ; Fuguang HUANG ; Yaping ZHAO ; Haiyan SUN ; Daozhu WU ; Xiuyun LI ; Shuying JIANG
Chinese Journal of Ultrasonography 2013;22(10):865-868
Objective To investigate the application of echocardiography in the diagnosis of neonatal ductus arteriosus aneurysm (DAA).Methods A total of 27 neonates with DAA were detected by echocardiography.Echocardiographic features in these cases were explored and outcome of these neonates were followed up.Results There were four crucial features in DAA detected by echocardiography:① A vessel bump was shown on the left side of the origination of left pulmonary artery(LPA) in the high parasternal short-axis view.②A horizontal shunt jet from the bump to the origination of LPA was shown when interrogate the top of the bump.③The bump joined the descending arota when interrogate it down and back orientately.A ‘V' shaped sign was shown when the aortic arch appeared on the right side and the bump on the left side.④A slow red flow signal was found in bump when the scale of color Doppler was cut down.The follow-up echocardiography showed spontaneous closure of the DAA when 3 months of age except one case lost.There were no abnormality in the figure and function in heart.Conclusions Echocardiography can be a valuable method for the diagnosis of neonatal ductus arteriosus aneurysm.Neonate with DAA have a good outcome.
9.Quantitative Analysis of Longitudinal Strain and Strain Rate of Left Ventricle in Normal Infants and Young Children
Hongju KOU ; Haiping ZHANG ; Liang WANG ; Fuguang HUANG ; Shuying JIANG ; Xiuyun LI
Chinese Journal of Medical Imaging 2013;(11):837-840
Purpose To quantitatively evaluate longitudinal strain and strain rate (SR) of left ventricular (LV) in normal infants and young children using velocity vector imaging (VVI). Materials and Methods A total of 50 normal infants and young children were enrolled in the study. The longitudinal strain peaks and SR peaks in three sections of LV walls during systole period and relaxation period were measured by VVI from apical four-chamber view, apical two-chamber view and long axis view, respectively. Results Forty-one cases (82%) presented a smooth strain curve without incisures and a double peak in SR curve (systolic peak and single diastole peak). The strain in anterior septum and posterior septum, the systolic SRs and diastolic SRs decreased from basal to apical segments. However, only the strain in basal segment and that in apical segment in anterior septum showed statistic difference (t=1.79, P<0.05);the systolic SRs of the same level in different walls and those in the same wall at different levels had no statistic difference (P>0.05). Conclusion Most normal infants and young children present a smooth diastolic strain curve and a double peak SR curve of LV, which can provide clinical guidance in the early diagnosis of myocardial abnormality for infants and young children.
10.Study on the approximal side angle of carotid plaques in patients with cerebral infarction using ultrasonography
Pintong HUANG ; Li WANG ; Xiaojing ZHANG ; Xinqiao TIAN ; Haiyan SUN ; Xiaotong WANG ; Chunpeng ZOU ; Fuguang HUANG
Chinese Journal of Ultrasonography 2009;18(12):1040-1042
Objective To assess the value of approximal side angle of carotid atherosclerotic plaques in patients with cerebral infarction by ultrasonography.Methods One hundred and seventy six patients with carotid atherosclerotic plaques were examined with uItrasonography and divided into two groups:cerebral infarction group (n=67)and none cerebral infarction group(n=109).Vessel area and residual lumen area of carotid artery were measured in axial view using linear probe.Area stenosis rate was calculated.The approximal side angle and thickness of plaque were measured respectively.The results of two groups were compared.Results The approximal side angle of plaque of cerebral group was significantly higher than that of none cerebral infarction groups(P<0.05).The sensitivity and specificity for approximal side angle of plaque(cutoff value:13.0°)were 79.1%,71.6%,respectively.There were no significant difference of area stenosis rate and plaque thickness between two groups(P>0.05).Conclusions The approximal side angle of plaque can be regarded as a novel index to assess the stability of plaque.