BACKGROUND: Reumatoid arthritis (RA) is a well-known autoimmune disease. Recently, polyglycosides of tripterygium wilfordii hook F (T Ⅱ), a traditional Chinese herb, has been widely used to treat rheumatoid arthritis.But the effects of TⅡ on the joints' synovium inflammation and whether TⅡcan prevent the reumatoid arthritis need to be investigated further.OBJECTIVE: To study the effects of T Ⅱ on the relative data of ankle joints of adjuvant arthritis (AA) rats.DESIGN: A randomized controlled experiment based on rats. SETTING: Departments of Histology and Embryology and Neurobiology, West China School of Preclinical and Forensic Medicine, Sichuan University. MATERIALS: A total of 20 healthy clean grade female SD rats, aged 2 to 3 months old, weighing 185-215 g, were provided by the Experimental Animal Center of West China Medical Center of Sichuan University. Fre und's complete adjuvant (FCA) was produced by Sigma Company. TⅡ was produced by Zhuzhou 3rd pharmacy of Hunan Province (certification number: 2005 No 055172, 10 mg/pill).METHODS: The experiment was completed in Department of Histology and Embryology and Neurobiology in Sichuan University from May 2004 to March 2005. ① All the 20 rats were randomly divided into 4 groups with 5 rats in each group by lots: normal group, without Freund's complete adjuvant (FCA) injection and TⅡ administration; model group, with FCA intradermal injection (0.2 mL) into the left hind paw and without TⅡ administration; TⅡ preventive group, first we use the same way as model group to replicate the AA model in rats, then on the 7th day AA rats were feed by TⅡ 30 mg/kg every day for 7 days; TⅡ therapeutic group, AA rats model were built with the same way as model group, on the 19th day AA rats were feed by TⅡ 30 mg/kg every day for 7 days. During this period, the swelling dimension of hind paw both primary and secondary are mea sured before immunization with FCA and after immunization, that was, on the 2nd, 10th, 15th, 19th, 22nd and 26th days. ② Arthritis index have been recorded according to inflammatory state of other three uninjected limbs.③ On the 28th day, all the rats were killed, the ankle joints are collected after perfusion-fixation. These joints were sectioned and colorated with H. E staining. Then we observe the histopathological changes in the synovium of ankle joints. MAIN OUTCOME MEASURES: Swelling dimension of joints and arthritis index, histopathology of anklebone joint's synovium. RESULTS: All of the 20 rats completed the experiment without missing. ① On the 2nd day after FCA injection, the primary hind paw of other three groups beside normal group appeared obvious swelling; from 2nd to 26th days, the volume of hind paw in other three groups was larger than that of normal group (t=2.315-3.041, P ＜ 0.05). The volume of primary hind paw in TⅡ preventive group at different time points (10th, 15th, 19th, 22nd and 26th days) was obviously less than that of model group (t=2.064-2.683, P ＜ 0.05). The volume of primary hind paw in TⅡ therapeutic group on the 22nd and 26th days was less than that of model group (t=2.112-2.578, P ＜ 0.05). Fifteen days after FCA injection, the volume of secondary hind paw in model group and T Ⅱ therapeutic group was larger than that of normal group (t=2.201-2.546, P ＜ 0.05). On the 15th, 19th, 22nd and 26th days, the volume of secondary hind paw in T Ⅱ preventive group was obviously less than that of model group (t=2.373-2.425, P ＜ 0.05). The volume of secondary hind paw in T Ⅱ therapeutic group on the 26th day was obviously less than model group (P ＜ 0.05). ② On the 14th day after FCA injection(after T Ⅱ preventive administration for 7 days), arthritis index of model group was (8.3±2.0) points, while arthritis index of TⅡ preventive group was (0.4±0.95) points (t=2.64, P ＜ 0.05), there was an obvious decline in T Ⅱ preventive group compared with model group. On the 26th day after FCA injection (after TⅡ therapeutic administration for 7 days), arthritis index of model group was (11.2±0.7), whileinflammatory disease in AA rats and prevent the secondary arthritis in the rats of AA as well.

Objective To investigate the effects of N - acetylcysteine (NAC) on apoptosis and the expressions of Fas/FasL mRNA in lung tissue of rats with paraquat - induced acute lung injury.Methods Forty five male SD rats were randomly (random number) divided into normal control group,paraquat (PQ) group,and NAC treatment group.The rat model of acute lung injury was made with 2％ PQ induction in dose of 25 mg/kg injected,and NAC was injected into the PQ poisoning rats (200 mg/kg) 30 minutes after PQ administration in NAC treatment group.In the control group,equal amount of saline instead was injected into the rats.Apoptosis was detected by using TUNEL method and the expressions of Fas/FasL mRNA were evaluated by using reverse transcription polymerase chain reaction (RT- PCR),and the levels of Fas/FasL protein were detected by using western blot analysis.Results Compared with control group,cell apoptosis and expressions Fas/FasL mRNA in PQ group were significantly different ( P ＜ 0.05 ).Compared with PQ group,cell apoptosis and expressions Fas/FasL mRNA in NAC group were significantly decreased,were significant lower (P ＜ 0.05).Conclusions NAC inhibited apoptosis in lung tissue of rats with paraquat induction by regulating the activation of Fas/FasL systems.

Objective To observe the expression of tumor necrosis factor-α(TNF-α)in acute lung injury caused by paraquat(PQ)in rats,and investigate the mechanism of the rhubarb in respect of pmteetive effects.Method PQ intragastrically poisoning at the dose of 50 mg/kg made a model of the acute lung injury in Sprague-Dawley(SD)rats.Totally 144 adult healthy SD rats(72 female,72male)were randomly divided into control group (group A,n=24),poisoned group(group B,n=48),rhubarb treated group(group C,n=48)and the shaln poisoning group(group D,n=24).Rats of group B and group C were poisoned intmgastrically with PQ(50 mg/kg).and rats of group C and group D were intervened intragastrieally with 300 mg/(kg·d)of rhubarb in 15 min-utes.The white blood cells and total cells in bronchoalveolar lavage fluid(BALF)were counted by using a blood cell counting plate and the protein content of BALF was measured by using the way of Lowry in order to calculate the neutmphiks pereentage and lung permeability index.A small portion of left lung was stained with HE to observe the pathological changes and the expression oftumor necrosis factor-α in the rest of the left lung was observed with immunohistochemistry.The data were handled by the analysis of variance and NK method using SPSS 14.0.Re-suits Compared with group A,the lungs of rats mainly showed congestion,edema and leukocytes infiltration in group B,and fibrosis was found onlyt in a few rats.And the rate neutrophils percentage,protein content and lung permeability index in BALF increased(P＜0.01).The expression of TNF-α were obviously inereased at 12 hours after PQ poisoning,and immtmohistochemistry score (IHS)was higher,and peaked at 24 hours later(P＜0.05),then remained on a high level for a while and sluggishly declined.Compared with group B,the changes of above mentioned were alleviated obviously,and the expression of TNF-α delayed with the less magnitude of increasing an an obvious tendency of less expression.Compared with group B,delayed,lower increasing extent,obviously re-ducing tendency in group C with statistical difference in IHS(P＜0.05).Conclusions Rhubarb ameliorates a-cute lung injury caused by PQ poisoning in rats by means of inhibiting the expression of TNF-α in turn to alleviate inflammatory reaction.

Objective To evaluate clinical significance of the grade of ischemia by QRS complex on the admission electrocardiogram(ECG)to predict severe arrithmia in patients with acute ST-segment elevation myocardial infarction(STEMI).Methods Patients with acute ST-segment elevation myocardial infarction(STEMI)admitted to emergency department from July 2003 to April 2008 were enrolled.A total of 223 patients met the criteria(ischemic chest pain ≥ 30 min,2 or more adjacent leads of ST segment elevation and onset time within 12 h).Exclusion criteria were bundle branch block and left ventricular hypertrophy.All enrolled patients were divided into two groups based on the enrollment electrocardiogram:grade 2 ischemia(ST elevation without terminal QRS distortion; n =134)and grade 3 ischemia(ST elevation with terminal QRS distortion; n =89).Patients of the two groups had comparable genderproportion,average age and coronary heart disease risk factors etc.All patients received thrombolytic therapy.The incidence rate of ST segment resolution(STR)and severe arrithmia in hospital stay were observed.Numerical variables were expressed mean ± standard deviation and compared by unpaired Student't test,Categorical variables were expressed percentage and compared by chi square test.Multiple logistic regression analysis was used to determine independent predictors of severe arrithmia.Results Patients with grade 3 ischemia had greater Σ ST on admission and 2 h after thrombolysis ECGs(P ＜ 0.01),the incidence rate of STR in patients with grade 3 ischemia was lower than that in patients with grade 2 ischemia(P ＜0.01).The peak creatine kinase MB fraction was higher in patients with grade 3 ischemia than that in patients with grade 2 ischemia(P ＜ 0.01).There was no significant difference of the incidence of severe arrithmia,such as ventricular premature beat,ventricular tachycardia or fibrillation,second-degree or third-degree atrioventricular block,and sinus arrest between the two groups(P ＞ 0.05),but there was a trend of higher incidence of severe arrithmia in patients with grade 3 ischemia compared with that in patients with grade 2 ischemia.Multiple logistic regression analysis demonstrated that the independent predictors of severe arrithmia were duration from symptom to thrombolysis and initial.Σ ST,whereas grade 3 ischemia remained a strong predictor of severe arrithmia.Conclusions Grade 3 ischemia on admission is associated with lower incidence of STR in patients with ST-segment elevation myocardial infarction(STEMI)after thrombolysis and a strong predictor of severe arrithmia.

Objective To investigate the effects of melatonin (MT) on rats with acute paraquat (PQ) poisoning. Method Fifty-four Sprague-Dawley (SD) rats were randomly divided into three groups (each group 18 rats) and given the following treatment: intragastric injection of PQ at 50 mg/kg (PQ); intragastric injection of paraquat followed by intraperitoneal injection of MT at 10mg/kg once a day (MT); intragastric injection of normal saline (Control). Serum assays for malondialdehyde (MDA) levels and superoxide dismutase (SOD) and glu tathione peroxidase (GSH-Px) activities were determined on the 1st, 3rd and 7th day post treatment. Clinical manifestations of poisoning and pathological changes in the lungs were also observed. Results Serum MDA levels were significantly increased (P ＜ 0.01), and SOD and GSH-Px activities significantly decreased (P ＜ 0.05) in the PQ group compared to the control group. Serum MDA levels were significantly decreased, and serum SOD and GSH-Px activities increased in MT group compared to the PQ group (P ＜ 0.05). Clinical manifestations of intoxication and pathological lung changes were also ameliorated in poisoned rats treated with MT. Condutions Administration of MT alleviates clinical manifestations of acute paraquat poisoning in rats by Limiting the damage from lipid peroxidation.

OBJECTIVETo investigate the effects of acetamide at different doses on the expression of inhibitory amino acids (gamma-aminobutyric acid, GABA) and excitatory amino acid (glutamate, Glu) in the cerebral cortex of rats with acute tetramine (TET) poisoning.

METHODSEighty Sprague-Dawley rats (SPF) were randomly divided into five groups, with 16 rats in each group: saline control group, dimethyl sulfoxide (DMSO) control group, TET exposure group, high-dose (2.8 g/kg/d) acetamide treatment group, and super-high-dose (5.6 g/kg/d) acetamide treatment group. Rats in the exposure group and treatment groups were exposed to TET by intragastric administration after fasting, and were then intramuscularly injected with saline or different doses of acetamide in the following 5 days. The cortex of the temporal lobe was collected at 3 h, 12 h, 48 h, or 7 d after treatment. The expression levels of GABA and Glu in the cortex of the temporal lobe were determined by average optical density (OD) values in immunohistochemistry.

RESULTS1) Expression of GABA: The OD value of GABA in TET exposure group started to increase at 12 h after treatment, reached the peak at 48 h, and decreased to the normal level at 7 d. In the high-dose acetamide treatment group, the increase in OD at 12 h was not so significant as that in the TET exposure group, OD value decreased to the normal level at 48 h and was lower than that in the exposure group, and the changes were more like those in the control groups. In the super-high-dose acetamide treatment group, OD value began to increase significantly at 3 h and was significantly higher than that in the TET exposure group (P < 0.01), it reached the peak at 12 h, and was restored to the normal value at 48 h. 2) Expression of Glu: The OD value of Glu in TET exposure group at 3 h after treatment was significantly lower than those in the two control groups, it increased gradually from 12 h to 48 h, and recovered to the normal level at the 7th d. The changes in the high-dose acetamide treatment group were similar to those in the TET exposure group, but became more like those in the control groups after 48 h; the OD value in super-high-dose acetamide treatment group was significantly higher than that in the TET exposure group at 3 h after treatment (P < 0.01), while no significant difference was found at 12 h; it was significantly lower than those of all other groups at 48 h and 7 d (P < 0.01).

CONCLUSIONSTreatment with high dose of acetamide has some curative effect on TET poisoning-induced central nervous lesion, while the effect of super-high-dose acetamide on expression of neurotransmitters is too complex to evaluate.

Acetamides ; pharmacology ; Animals ; Bridged-Ring Compounds ; poisoning ; Cerebral Cortex ; drug effects ; metabolism ; Female ; Glutamic Acid ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; gamma-Aminobutyric Acid ; metabolism

OBJECTIVETo observe the effect of different doses of acetamide on the histopathology in the cerebral cortex of rats with tetramine (TET) poisoning and to provide a basis for the treatment of fluoroacetamide poisoning with acetamide.

METHODSEighty clean Sprague-Dawley rats were randomly divided into five groups: saline control group,dimethylsulfoxide water solution control group,TET poisoning group, acetamide (2.88 g/kg/d) treatment group, and acetamide (5.68 g/kg/d) treatment group, with 16 rats in each group. Rats in the poisoning group and treatment groups were poisoned with TET by intragastric administration after fasting; then, saline was injected intramuscularly into rats of the poisoning group, and different doses of acetamide were injected intramuscularly into rats of treatment groups; the course of treatment was 5 d. At 3 h, 12 h, 48 h, and 7 d after treatment, the cerebral cortex was harvested from rats in each group, and the histopathological changes in the cerebral cortex were evaluated under light and electron microscopes.

RESULTSThe light microscopy showed that the TET poisoning group had hypoxia changes in the cerebral cortex, which worsened over time; the treatment groups had reduced hypoxia changes, and the acetamide (2.88 g/kg/d) treatment group had more reduction than the acetamide (5.68 g/kg/d) treatment group. The electron microscopy showed that the apoptosis of neuronal cells were the main pathological changes in the TET poisoning group; the treatment groups had reduced apoptotic changes, and the acetamide (2.88 g/kg/d) treatment group had more reduction than the acetamide (5.68 g/kg/d) treatment group.

CONCLUSIONNo pathological changes associated with the synergistic toxic effect of acetamide and TET are found in the cerebral cortex. Acetamide (2.88 g/kg/d) could reduce central nervous lesions, but the efficacy is not improved after increasing the dose. For patients who cannot be identified with TET or fluoroacetamide poisoning, acetamide could be considered for treatment.

Acetamides ; pharmacology ; Animals ; Bridged-Ring Compounds ; toxicity ; Cerebral Cortex ; drug effects ; pathology ; Disease Models, Animal ; Male ; Rats ; Rats, Sprague-Dawley

Gut microbiota dysbiosis is an avenue for the promotion of atherosclerosis (AS) and this effect is mediated partly via the circulating microbial metabolites. More microbial metabolites related to AS vascular inflammation, and the mechanisms involved need to be clarified urgently. Paeonol (Pae) is an active compound isolated from Paeonia suffruticoas Andr. with anti-AS inflammation effect. However, considering the low oral bioavailability of Pae, it is worth exploring the mechanism by which Pae reduces the harmful metabolites of the gut microbiota to alleviate AS. In this study, ApoE^-/- mice were fed a high-fat diet (HFD) to establish an AS model. AS mice were administrated with Pae (200 or 400 mg·kg^-1) by oral gavage and fecal microbiota transplantation (FMT) was conducted. 16S rDNA sequencing was performed to investigate the composition of the gut microbiota, while metabolomics analysis was used to identify the metabolites in serum and cecal contents. The results indicated that Pae significantly improved AS by regulating gut microbiota composition and microbiota metabolic profile in AS mice. We also identified α-hydroxyisobutyric acid (HIBA) as a harmful microbial metabolite reduced by Pae. HIBA supplementation in drinking water promoted AS inflammation in AS mice. Furthermore, vascular endothelial cells (VECs) were cultured and stimulated by HIBA. We verified that HIBA stimulation increased intracellular ROS levels, thereby inducing VEC inflammation via the TXNIP/NLRP3 pathway. In sum, Pae reduces the production of the microbial metabolite HIBA, thus alleviating the ROS/TXNIP/NLRP3 pathway-mediated endothelial inflammation in AS. Our study innovatively confirms the mechanism by which Pae reduces the harmful metabolites of gut microbiota to alleviate AS and proposes HIBA as a potential biomarker for AS clinical judgment.
Animals ; Mice ; Atherosclerosis/drug therapy* ; Diet, High-Fat ; Endothelial Cells ; Inflammation/drug therapy* ; Mice, Inbred C57BL ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Reactive Oxygen Species