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Objective:Using atenolol as a model drug,the aim of this study was to develop a sustained and controlled transdermal drug delivery system(TDDS)based on polyethyleneimine-modified MoS2 nanoparticles(PEI-MoS2 NPs)that were responsive to near infrared(NIR)laser irradiation.Methods:The three-dimensional flower-like PEI-MoS2 NPs were successfully synthesized and further characterized by attenuated total reflection Fourier transform infrared spectroscopy,X-ray diffraction measurements,scanning electron microscopy,and transmission electron microscopy.The controlled release capacity of PEI-MoS2 NPs was examined using in vitro drug release and skin penetration experiments.Results:The PEI-MoS2 NPs exhibited a drug loading efficiency of 53.86% and high photothermal conversion ability.Moreover,the release of atenolol was enhanced by NIR stimulation with an enhancement ratio of 1.56.Conclusion:NIR-controlled PEI-MoS2 NPs was essential for the control and sustained release of drugs in TDDS.
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Objective To investigate the changes and formation mechanism of plasma endothelial microparticles (EMPs) in patients with acute pancreatitis (AP). Methods Blood samples were collected from 60 patients with AP who were treated in The First Affiliated Hospital of Anhui Medical University from August 2020 to June 2021, and these patients were divided into mild acute pancreatitis (MAP) group with 23 patients, moderate-severe acute pancreatitis (MSAP) group with 23 patients, and severe acute pancreatitis (SAP) group with 14 patients; 20 individuals who underwent physical examination were enrolled as control group.Differential centrifugation was used to obtain platelet-poor plasma, flow cytometry was used to measure the level of CD31 + CD41 - EMPs, and ELISA was used to measure the levels of endothelin-1(ET-1), von Willebrand factor (vWF), nitric oxide (NO), and vascular cell adhesion molecule-1(VCAM-1).HUVECs were stimulated by the plasma of AP patients, and then flow cytometry and qRT-PCR were used to measure the changes in EMPs, reactive oxygen species (ROS), and mitochondrial membrane potential and the expression of endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS), intercellular adhesion molecule-1(ICAM-1), VCAM-1, NADPH oxidase, and P-selectin.A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups.The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between groups and within each group.The chi-square test was used for comparison of categorical data between groups, and the Pearson correlation test was used for correlation analysis. Results Compared with the control group, the MAP, MSAP, and SAP groups had a significant increase in the level of EMPs (all P < 0.05).Compared with the MAP and MSAP groups, the SAP group had a significant increase in the level of EMPs (both P < 0.05).In the patients with AP, the level of EMPs was negatively correlated with Acute Physiology and Chronic Health Evaluation Ⅱ score, Bedside Index for Severity in Acute Pancreatitis, Ranson score, CT score, and C-reactive protein ( r =0.686 2, 0.777 3, 0.713 8, 0.771 8, and 0.473 9, all P < 0.01).Compared with the control group, the MAP, MSAP, and SAP groups had significant increases in the levels of ET-1, vWF, and VCAM-1 and a significant reduction in the level of NO (all P < 0.05).Compared with the control group, the MSAP and SAP groups had the plasma that promoted the release of a large amount of EMPs (both P < 0.05).Compared with the control group, all the other groups, except the MAP group in terms of VCAM-1 and eNOS, had significant increases in the mRNA expression levels of eNOS, iNOS, ICAM-1, P-selectin, VCAM-1, and NADPH oxidase (all P < 0.05).Compared with the HC group, the MAP, MSAP, and SAP groups and the LPS group had a significant increase in the level of ROS and a significant reduction in mitochondrial membrane potential in HUVECs (all P < 0.05). Conclusion There is a significant increase in the plasma level of EMPs in AP patients, which is correlated with the severity of pancreatitis.Meanwhile, the plasma of AP patients can promote the formation of EMPs in HUVECs in vitro, which may be associated with cell oxidative injury.
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Objective:To understand the epidemiological characteristics and pathogenic spectrum of influenza-like illnesses in Tianjin Children′s Hospital from October 2020 to March 2021, and to provide reference for the prevention, control and clinical diagnosis and treatment of influenza-like illnesses.Methods:A total of 520 throat swabs samples were collected from patients with influenza-like illnesses in sentinel hospitals. Thirty respiratory tract pathogens were detected by real-time fluorescence quantitative PCR. The results were statistically analyzed by descriptive epidemiological methods.Results:Among the 520 samples, 239 were positive for 16 respiratory pathogens with a positive rate of 45.96%. The top three pathogens were respiratory syncytial virus (9.62%, 50/520), rhinovirus (9.62%, 50/520) and cytomegalovirus (5.96%, 31/520). The positive rate of respiratory pathogens was 49.67% in males and 40.91% in females and the difference between males and females was statistically significant (χ 2=3.919, P<0.05). There were significant differences in the positive rates among three age groups (χ 2=6.182, P<0.05) with the highest positive rate in the <2 years old group (52.91%, 91/172) and the lowest rate in the >4 years old group (38.10%, 40/105). There were significant differences in the positive rates detected in different months (χ 2=15.358, P<0.05) and the highest detection rate was in December (58.00%, 58/100), followed by those in November (52.50%, 42/80) and January (47.50%, 38/80). The multiple infection rate was 21.76% (52/239) and most of the multiple infections were caused by rhinovirus and other pathogens (48.08%, 25/52). Conclusions:Respiratory syncytial virus, rhinovirus and cytomegalovirus were the predonimant pathogens responsible for influenza-like illnesses in Tianjin Children′s Hospital from October 2020 to March 2021. Multiple infections were common and children under 2 years old were more susceptible. The detection rate of respiratory pathogens varied in different months. It was necessary to strengthen the surveillance and research on those respiratory pathogens in order to provide scientific data for the prevention and control of respiratory diseases in children.
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Objective:To retrospectively analyze the test results of novel coronavirus (2019-nCoV) in different samples (throat swab, sputum and feces) collected from recovered COVID-19 patients in order to provide a more reliable basis for discharge and reduce the risk of recurrence after discharge.Methods:Throat swabs and sputum were sampled in pairs from 78 patients before discharge and sampled in pairs twice from 54 cases with an interval of 1-5 d. Real-time fluorescence quantitative RT-PCR was used to detect the virus in the two types of samples. Throat swab, sputum and fecal samples of six patients were tested for 2019-nCoV during follow-up.Results:The detection rate of viral nucleic acid was 46.15% in throat swabs and 50.00% in sputum samples. Test results of the second paired samples showed that the detection rate of viral nucleic acid was 25.93% in throat swabs and 46.30% in sputum samples, and the difference between the two types of samples was statistically significant ( P<0.05). During follow-up, 2019-nCoV nucleic acid could be detected in the fecal samples of the six patients, but not in their throat swab and sputum samples. Their fecal samples remained positive up to 52 d. Conclusions:In the late convalescence, the respiratory symptoms of COVID-19 patients gradually disappeared with the improvement of clinical symptoms. Moreover, the virus might enter the gastrointestinal tract from respiratory tract, and could long-term exist in recovered patients and be excreted in feces. In order to reduce the rate of missed detection and avoid false negative results, it was suggested to test the viral nucleic acid in different types of samples before a COVID-19 patient was discharged.
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Objective To analyze the clinical characteristics, treatments and prognosis of acute obstructive suppurative pancreatic ductitis ( AOSPD) , and to discuss its pathogenesis, diagnosis and treatment strategy. Methods 63 AOSPD cases reported in Chinese and foreign literature from June 1993 to January 2019 were collected. The sex, age of onset, etiology and potential risk factors, clinical manifestations, laboratory examinations, imaging findings, treatments and prognosis were recorded. Results The male to female ratio was 53 / 10, and the median age of onset was 59 years. The etiology and risk factors included chronic pancreatitis, pancreatic neoplasms, diabetes mellitus, history of endoscopic intervention and alcoholism before the onset of AOSPD. The main clinical manifestations were epigastric pain and fever, and sepsis and shock might occur in a few cases. The serum amylase was 13-1946 ( IU/L) at the early stage of onset and it decreased to varying degrees after treatments. Imaging examination showed that pancreatic duct dilatation was found in 54 patients and pancreatic duct stones were found in 42 patients. Pancreatic juice culture was bacteria-positive in more than 31 cases, and the common pathogenic bacteria were Enterococcus and Escherichia coli. Therapeutic methods included endoscopic pancreatic stent implantation ( n=36 ) , endoscopic nasopancreatic drainage (n=22), surgical operation (n=4) and antibiotic treatment, and the condictions in most of the patients were improved to some extent after treatments. Conclusions Older age, male, chronic pancreatic disease, history of endoscopic intervention and drinking, and diabetes mellitus were the main etiological factors of AOSPD. The clinical manifestations of AOSPD were nonspecific but could be complicated by severe complications. Imaging examination and pancreatic juice culture can help to confirm the diagnosis. Antibiotic therapy, timely endoscopic interventions and surgical procedures can improve the short-term prognosis.
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Objective@#To compare the safety and immunogenicity of two different sequential schedules of inactivated poliomyelitis vaccine made from Sabin strain (sIPV) followed by typeⅠ+Ⅲ bivalent oral poliovirus vaccine (bOPV) in Drug Candy (DC) form or liquid dosage form).@*Methods@#This randomized, blinded, single center, parallel-group controlled trial was done from September 2015 to June 2016 in Liuzhou, Guangxi province. Healthy infants aged ≥2 months were eligible for enrollment and divided into 1sIPV+2bOPV or 2sIPV+1bOPV sequential schedules. According to the bOPV dosage form each sequential schedules, the subjects again were divided into drug candy(DC) form or liquid dosage form group, being 1sIPV+bOPV (DC)/1sIPV+2bOPV(liquid)/2sIPV+1bOPV(DC)/2sIPV+1bOPV(liquid). According to 0, 28, 56 d immunization schedule, Each group were given 3 doses. We recorded adverse events during the clinical trial (399 participants who receive at least one dose). 28 days post-Dose 3, we receive a total of 350 blood samples (excluding the quitters or subjects against trial plan), using cell culture trace against polio virus neutralization test Ⅰ, Ⅱ, Ⅲ neutralizing antibody (GMT), calculating the antibody positive rate.PolioⅠ,Ⅱand Ⅲ antibody titers were assessed by virus-neutralizing antibody assay and the seroconversion (4-fold increase in titer) from pre-Dose 1 to 28 days post-Dose 3 was calculated (total 350 samples) .@*Results@#During the vaccination, the incidence of AEs in 1sIPV+2bOPV(DC), 1sIPV+2bOPV (liquid), 2sIPV+1bOPV(DC), 2sIPV+1bOPV (liquid) group were 79%, 76%, 80% and 74% (χ2=1.23, P=0.747) , respectively. The severe AEs in groups were 6%, 5%, 6% and 4% (χ2=0.57, P=0.903) , respectively, and none was considered to be vaccination related. 28 days after 3rd vaccination, the seroconversion rates in 1sIPV+2bOPV (DC), 1sIPV+2bOPV (liquid), 2sIPV+1bOPV (DC), 2sIPV+1bOPV (liquid) group, were 99%, 100%, 99% and 99% (χ2=0.94, P=0.815) , respectively, for type Ⅰ poliovirus; and 47%, 57%, 80%, 79% (χ2=31.56, P<0.001) , respectively, for type Ⅱ; and were 100%, 99%, 100%, 99% (χ2=2.02, P=0.568) , respectively, for type Ⅲ. In each group, the GMT of antibody against poliovirus typeⅠ were 4 539.68, 6 243.43, 6 819.53 and 7 916.29 (F=25.87, P<0.001) , respectively; Type Ⅱ were 12.98, 10.54, 63.75 and 84.21 (F=8.68, P=0.034) , respectively; Type Ⅲ were 1 172.55, 1 416.03, 2 648.89 and 3 250.75 (F=14.50, P=0.002) , respectively.@*Conclusion@#On the same sequential schedules, there was no significant difference between the dosage forms, all of them showed good safety and immunogenicity. In the same dosage forms with different sequential schedules, the seroconversion rate was higher in 2 dose sIPV group than the 1 dose sIPV group, especially at the neutralizing antibody GMT level against polio type Ⅱ and Ⅲ after vaccination.
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Objective@#To investigate the effects of resolvin D1 on autophagy in the prevention of acute pancreatitis (AP) in mice. @*Methods@#Thirty C57BL/6 mice were divided into control group, AP group and resolvin D1 group. AP model was established by intraperitoneal injection of cerulein at 50 μg·kg-1·h-1. Resolvin D1 was intraperitoneally given at 50 μg/kg one hour before and four hours after modeling. The mice of control group were intraperitoneally injected the same volume of 0.9% sodium chloride solution. The serum levels of amylase and lipase were measured by colorimetric method. The pathological injury of the lung and pancreatitis were observed under optical microscope. Autophagic vacuoles in acinar cells of pancreas of mice were evaluated by transmission electron microscope. And the expressions of autophagy related markers Beclin-1, p62 and LC3-Ⅱ at the mRNA and protein levels in pancreas of mice were detected by real time quantitative polymerase chain reaction (RT-qPCR) and Western blotting method. One-way analysis of variance and SNK-q were performed for statistical analysis. @*Results@#There were statistically significant differences in serum amylase and lipase levels between control group, AP group and resolvin D1 group (F=62.99 and 149.69, both P<0.01). The serum amylase and lipase levels of mice in resolvin D1 group were lower than those of AP group ((525.08±41.12) U/L vs. (752.62±42.03) U/L, (758.24±134.77) U/L vs. (1 201.06±112.53) U/L), and the differences were both statistically significant (both SNK-q test and P<0.01). In addition, there were statistically significant differences in the ratio of the pancreas and lung wet mass to body mass between control group, AP group and resolvin D1 group (F=11.36 and 18.51, both P<0.05). Pathological injury scores of pancreas and lung of resolvin D1 group were both lower than those of AP group (3.3±0.6 vs. 5.6±0.6, 5.4±0.5 vs. 8.8±0.4), and the differences were statistically significant (both SNK-q test and P<0.05). The results of transmission electron microscopy observation revealed that the number of autophagic vacuole of resolvin D1 group was less than that of AP group, and the size was smaller. Moreover, there were statistically significant differences in Beclin-1, p62 and LC3-Ⅱ mRNA between control group, AP group and resolvin D1 group (F=270.95, 151.83 and 124.77, all P<0.05). The relative expression mRNA levels of Beclin-1, p62 and LC3-Ⅱ of resolvin D1 group were 1.59±0.12, 2.75±0.27 and 1.34±0.14, respectively, which were lower than those of AP group (2.68±0.13, 3.32±0.30 and 3.37±0.26, respectively), and the differences were statistically significant (all SNK-q test and P<0.05). There were statistically significant differences in the expressions of Beclin-1, p62 and LC3-Ⅱat the protein level between control group, AP group and resolvin D1 group (F=116.63, 384.40 and 192.45, all P<0.05). The expressions of Beclin-1, p62 and LC3-Ⅱ at protein level of resolvin D1 group were 0.98±0.03, 0.57±0.04 and 0.31±0.03, respectively, which were lower than those of AP group (1.34±0.07, 1.02±0.03 and 0.48±0.04, respectively), and the differences were statistically significant (all SNK-q test and P<0.05). @*Conclusion@#Resolvin D1 ameliorates the severity of AP by attenuating the impaired autophagy and restoring autophagic flux in AP mice.
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Aim To investigate whether Hcy influenced the intestinal mucosal permeability by regulating MEK-ERK-MLCK pathway. Methods SD rats were divided into 4 groups:normal group, normal+Hcy group, TN-BS/ethanol group, TNBS/ethanol+Hcy group. Experi-mental colitis model with hyperhomocystinemia was es-tablished in rats with intracolonic administration of TN-BS and subcutaneous injection of Hcy. The colonic mucosal tissue was collected for histopathological exam-ination and activity of myeloperoxidase ( MPO ) . The protein expression of MLCK, p-MLCK, MEK, ERK and p-ERK in intestinal mucosal tissues was examined by Western blot method. The mRNA expression of ML-CK was examined by RT-qPCR method. Result Com-pared with the normal group and TNBS group, the DAI and HI scores and the MPO activity were increased in TNBS/ethanol+Hcy group ( P <0. 01 ) . Western blot and RT-qPCR showed that expression of MLCK, p-ML-CK, MEK, ERK and p-ERK increased in small intes-tine in TNBS/ethanol+Hcy group. Conclusion Hcy can increase intestinal permeability in TNBS-induced colitis rats by regulating the expression of MEK-ERK-MLCK signal pathway.
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BACKGROUND:Distraction osteogenesis is one of the most important tissue engineering technologies. However, the exact signaling pathway controling mesenchymal stem cel-osteoblast lineage (MSC-OB) migration during distraction osteogenesis has not yet been elucidated. More efforts should be paid to make a ful understanding of the mechanism on MSC-OB lineage migration, which can improve the clinical efficacy of distraction osteogenesis. OBJECTIVE:To evaluate the effects of mechanical stretch on the ability of MSC-OB mobility and expression of mammalian target of rapamycin (mTOR) signaling pathway as wel as matrix metaloproteinases (MMPs) in MSC-OB, and to make clear the mechanism by which controls MSC-OB migration during distraction osteogenesis. METHODS:Twelve Sprague-Dawley rats were randomized into two groups: experimental group (n=6), anin vivo rat mandibular distraction osteogenesis model was established on the right side of rats; non-stretch group (n=6), only the mandibular resection was done but with no distraction osteogenesis. Immunohistochemical staining was used to detect phosphorylated mTOR expression in new osteotylus at 15 days after operation. In addition, an in vitro cel stretch model was made in the mandibular mesenchymal stem cels from healthy Sprague-Dawley rats under resting tension force (6%, 4 hours); no distraction was done in control group. The ability of MSC-OB mobility, the expression of mTOR, Raptor, p70S6K and MMPs were evaluated using experiment methods including immunohistochemistry staining, real-time PCR and scratch assay. RESULTS AND CONCLUSION: The expression of phosphorylated mTOR in MSC-OB was upregulated in the mandibular bone calus of the stretch group than the non-stretch group (P < 0.05). In thein vitro experiments, MSC-OB applied with mechanical stretch (6%, 4 hours) showed elevated gene expression levels of mTOR, Raptor, p70S6K, MMP-2, MMP-9 and MMP-13 compared with the control group (0%, 4 hours). Meanwhile, MSC-OB in the experiment group (6%, 4 hours) showed a greater ability of mobility, as demonstrated by a farther distance after 48 hours of observation (P < 0.05). The present study suggests that the enhancement of MSC-OB mobility correlates with increase of the gene expression of MMPs and mTOR signaling pathway. Mechanical stretch may promote MSC-OB migration through activation of mTOR/MMPs signaling pathway.
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Objective To explore the clinical characteristics and medicine treatment of patients with severe ulcerative colitis (UC),the efficacy of rescue treatment in patients with glucocorticoid (GCS) resistant severe UC,and the clinical risk factors in patients with GCS-refractory severe UC.Methods From January 2001 to December 2012,clinical,laboratory,endoscopy,imaging data and medication of treatment of 106 patients with severe UC were retrospectively analyzed.Then the patients were followed up,and the clinical efficacy and under endoscopic presentation of Mayo score were evaluated.Logistic regression analysis was performed to analyze the high risk factor of GCS-refractory severe UC.Results Among 106 patients with severe UC,95 were chronic relapse type accounting for 89.6 ％.The percentage of patients with defecation times over six was 73.6％ (78/106),with severe purulent bloody stool was 51.0％ (54/106),and with moderately or severe abdominal pain was 83.0％ (88/106).The percentage of diffuse colon type was 83.0％ (88/106),endoscopic presentation of Mayo score over two was 87.7％ (93/106).Hemoglobin decreased in 65.1％ (69/106) patients,blood platelet increased in 48.1％ (51/106) patients,C-reaction protein elevated in 88.7％ (94/106) patients,and hypoalbuminemia decreased in 42.5％ (45/106) patients.Account to 89.6％ (95/106) of patients with severe UC received GCS treatment,and the percentage of induced remission was 64.2％ (61/95),effective rate was 16.8％ (16/95),and ineffective rate was 18.9％ (18/95).The percentage of GCS refractory was 35.8％(34/95).There were 23 patients with GCS resistance and 11 patients with GCS dependence.Ten patients with GCS resistant severe UC accepted medicine rescue therapy.Five cases were treated with cyclosporin A,of which two cases induced remission,one case was effective,and two cases were ineffective.Another five cases were treated with infliximab,of which three cases induced remission,and two cases were ineffective.The results of Logistic regression analysis showed that severe anemia (OR=6.750,95％CI:2.656 to 17.152,P＜0.01),elevated blood platelets (OR=4.032,95％CI:1.226 to 13.261,P=0.015) and albumin level less than 25 g/L (OR =3.022,95 ％ CI:1.236 to 7.390,P =0.022) were risk factors of GCS-refractory severe UC.Conclusions GCS resistant or dependent occurred in part of patients with severe UC.Patients with G-CS resistant severe UC receive rescue treatment of cyclosporin A or infliximab.Severe anemia,elevated blood platelets,albumin less than 25 g/L may be clinical predicting factors in patients with GCS-refractory severe UC.
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BACKGROUND:Central nerve damage and peripheral nerve injury are common clinical problems that have no ideal treatment. Nerve growth factor has an important role in neuronal repairing and growth. But its local injections may have shorts of inactivation and loss. OBJECTIVE:To construct human nerve growth factor beta recombinant plasmids, which are transfected into bone marrow mesenchymal stem cells from the rabbit mandible by lentiviral vectors, and to investigate the bioactivity of human nerve growth factor beta. METHODS:pDC316-hNGFβ-mCMV-EGFP plasmids were constructed via lentiviral vectors using Hind III+Not I digestion. Bone marrow mesenchymal stem cells from the rabbit mandible were isolated and cultured, and then transfected by recombinant plasmids. The expression of human nerve growth factor beta in transfected cells was detected by ELISA method. RESULTS AND CONCLUSION:pDC316-hNGFβ-mCMV-EGFP plasmids were proved to be constructed successful y by gene sequencing and enzyme identification. The transfected cells under a fluorescence microscope emitted green fluorescence, and the fluorescence intensity had no change with incubation time. The expression of human nerve growth factor beta was maintained at a level of 25μg/L at 7 days after celltransfection, and the bioacitivty was increased significnalty.
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Objective To investigate the association between MYO9B rs962917 and rs1545620 gene polymorphism and clinical pathological characteristics of patients with inflammatory bowel disease (IBD) and permeability of intestinal mucosa.Methods From September 2010 to May 2012,a total of 196 cases of patients with IBD were collected,100 cases were ulcerative colitis (UC) and 96 cases were Crohn's disease (CD).At the same time,99 gender and age matched healthy individuals were collected as healthy controls.The 5 mL blood of participants was obtained and DNA was extracted.The MYO9B gene rs962917 and rs1545620 polymorphism was detected by polymerase chain reaction (PCR) and ligase detection reaction (LDR).After 60 patients with UC and 58 patients with CD orally took intestinal permeability testing fluid (with lactulose and mannitol),the urine of the patients was analyzed with high pressure liquid chromatography-pulsed lectrochemical dection (HPLC-PED).The permeability of intestinal mucosa was determined according to the ratio of lactulose and mannitol.Chisquare test was used for count data.Results Compared with healthy control group,there was no significant difference in genotype and allelic gene distribution of MYO9B rs962917 and rs1545620 of IBD group,UC group and CD group (all P＞0.05).The genotype of MYO9B rs962917 and rs1545620 of patients with UC was not related with the disease activity and location of lesions (rs962917:x2 =0.481 and 3.812,rs1545620..x2 =0.398 and 4.543 ;all P＞0.05).The genotype of MYO9B rs962917 of patients with CD was not related with the disease activity,lesion type and occurrence of perianal lesions (x2 =0.384,0.476 and 3.486,all P＞0.05) and was related with location of lesions (x2=15.926,P＜0.05).The genotype of MYO9B rs1545620 of patients with CD was not related with the disease activity and lesion type (x2 =1.407 and 5.126,both P＞0.05),however was related with location of lesions and occurrence of perianal lesions (x2 =18.165 and 7.629,both P＜0.05).The permeability of intestinal mucosa of all 58 patients with CD was high.The genotype of MYO9B rs962917 and rs1545620 of patients with UC was not related with the permeability of intestinal mucosa (x2=1.508 and 1.025,both P ＞ 0.05).Conclusion MYO9B rs962917 and rs1545620 gene polymorphism is related with the location of lesions in CD and is not related with the permeability of intestinal mucosa of patients with UC.
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Objective To investigate the effect and mechanism of anti-tumor necrosis factor (TNF)-α on the intestinal mucosal permeability in dextran sulfate sodium (DSS) induced colitis mice.Methods Eighteen C57BL/6J mice were evenly divided into healthy control group,model group and anti-TNF-α treated group.The mice of model group and anti-TNF-α treated group were fed with 5％DSS solution for 7 days.The mice of anti-TNF-α treated group were injected anti-TNF-α (5 mg/kg)intraperitoneally on the first and fourth day; control group and model group were substituted with equal volume saline injection.The mice were sacrificed at 7 days after modeling.The disease activity index (DAI) score was evaluated everyday.The intestinal permeability was examined by Evan′s blue (EB) method and FITC-dextran (FITC-D) method.The colon tissue was collected for observation under microscope and histological index (HI).The small intestinal tissues were examined under electron microscope.The 10％ homogenate of colon and intestinal mucosa was prepared,the activity of myeloperoxidase (MPO),the content of TNF-α and epithelial myosin light chain kinase (MLCK) concentration were determined with kits respectively.The expression of MLCK in intestinal mucosa was tested by Western blot assay.Single factor of variance between groups were analyzed.Results Compared with control group,the DAI of model group increased daily.Compared with model group,the DAI of anti-TNF-α treated group improved.In model group,mice intestinal epithelial cells junctional complex shortened and widened and the cell gap expanded.In anti-TNF-α treated group,the connection structure of mice intestinal epithelial cells was tighter.The activity of HI and MPO and the content of TNF-α of model group were higher than those of control group (P ＝ 0.008,0.006 and 0.001,respectively),all of those of anti-TNF-α treated group were lower than those of model group (P=0.004,0.008 and 0.005,respectively).The F value of three groups was 131.98,218.28 and 58.93,respectively.The contents of EB in mice intestinal wall and serum FITC-D of model group were higher than those of control group (P=0.003 and 0.010),and those of anti-TNF-α treated group were lower model group (P＝0.001 and 0.009).The F value of three groups was 69.36 and 17.96.The MLCK concentration in mice intestinal mucosa of model group [(71.10± 7.52) ng/g] was higher than that of control group [(18.56±9.92) ng/g,P＜0.01],that of anti-TNF-α treated group [(37.56±15.84) ng/g] was lower than model group (P=0.008),and the difference among these three groups was statistically significant (F＝ 17.23).The Western blot results indicated the expression of MLCK in intestinal mucosa of model group was higher than that of control group,and that of anti-TNF-α treated group was lower than model group.Conclusions Anti-TNF-α play an important role in improving colitis,and the intestinal mucosal permeability.The mechanism may be related with the regulation of MLCK expression.
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Objective To explore the clinical correlation of the variation of plasma homocysteine (HCY), melatonin (MLT) and ulceative colitis (UC). Methods The clinical data of 112 UC patients was collected, and 110 normal healthy persons as control. The level of plasma HCY and MLT was detected by high pressure liquid chromatography-fluorescence detection (HPLC-FD) method. The level of plasma folate ( FA) and vitamin B12 was detected by enzyme-linked immunosorbent assay (ELISA) method. The correlation of these four indexes and UC was analyzed. Results The serum level of HCY in UC patients was significantly higher than that in normal healthy persons [(11. 27± 7.26) μmol/L vs (8. 19±4. 81) μmol/L, P = 0. 000]. The serum level of MLT in UC patients was significantly lower than that in normal healthy persons [(49. 06 + 31. 40) pg/ml vs (64. 28±41. 16) pg/ml,P=0. 008]. The serum level of FA in UC patients was significantly lower than that in normal healthy persons [(7. 64 + 1.95) nmol/L vs (9. 14 + 1.23) nmol/L, P = 0. 005]. The serum level of vitamin B12 in UC patients was significantly lower than that in normal healthy persons [(108. 64 ±32. 22) pmol/L vs (112. 64±33. 33) pmol/L, P = 0. 004]. There was no correlation between plasma HCY, MLT and UC disease activity degree, range, disease duration, erythrocyte sedimentation rate (ESR), or C reactive protein (CRP) in UC patients. There was no significant correlation between MLT and HCY in UC patients. Conclusions The serum level of HCY is higher in UC patients than that in normal control, and MLT is lower than that in normal control. However there is no significant correlation between them.
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Objective To investigate the relationship between TNF-α, IFN-γand intestinal muco-sal permeability in a mouse colitis model and its inhibiting effect by balsalazide. Methods Forty-five C57BL/6J mice were divided randomly into five groups. Normal group was only fed with distilled water, DSS group and balsalazide groups at doses of 42, 141,423 mg/kg were both fed with 5% DSS. Balsalazide was given by intragastric administration. At the end of the experiment, colon tissue was collected for assessment of histological index(HI) and the MPO activity. Small intestinal mucosa was collected for assessment of the content of TNF-α and IFN-γ,transmission electron microscope(TEM), and detection of permeability by Ev-arts blue method. Results Compared with normal group, DSS group mice all manifested severe weight loss associated with hematocbezia and diarrhea, HI score, and the colon MPO activity and the content of TNF-α and IFN-γ were increased significantly. Intestinal mucosa showed a thinning of microvillous carpet, with de-curtated and broaden junctional complex and enlarged intercollutar space under TEM observations. The amount of Evans blue permeated into intestinal wall was obvious. Compared with DSS group, the HI score, the MPO activity and the content of TNF-α and IFN-γ were decreased by balsalazide. The amount of Evans blue permeated into intestinal wall was less. Ileal microvillous carpet was ameliorated dose dependently by balsalazide. Conclusion In DSS-induced colitis model, the change of the content of the TNF-α and IFN-γ, was accordance with the increase of intestinal mucosal permeability while balsalazide can significantly amelio-rate intestinal mucosal permeability by its anti-colitis effect.