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Objective:To observe the protective effect of forsythiaside A on acute lung injury (ALI) in septic rats.Methods:Male Sprague-Dawley (SD) rats were randomly divided into normal control group, sham operation group, sepsis model group, and forsythiaside A intervention group, with 10 rats in each group. The rats in the normal control group did not receive any intervention; the rats in the sham operation group only underwent abdominal surgery; and those in the model group and forsythiaside A intervention group received cecal ligation and puncture (CLP) to establish the sepsis rat model. The rats in the forsythiaside A intervention group were given 75 mL/kg of forsythiaside A within 0.5 hour after operation, and repeated after 6 hours. The rats in the sham operation group and model group were given the same amount of normal saline at the same time points. The lung tissues were collected for pathological examination 12 hours after operation. The lung homogenate was prepared, and enzyme-linked immunosorbent assay (ELISA) was used to detect tumor necrosis factor-α (TNF-α), interleukins (IL-1β, IL-6). The activity of superoxide dismutase (SOD) was detected by xanthine oxidase method, and the content of malonaldehyde (MDA) was detected by colorimetry. The expression of nuclear factor-κB p65 (NF-κB p65) was detected by Western blotting.Results:There was no significant pathological change of lung tissue in both normal control group and sham operation group, and there was no significant difference in each parameter between the two groups. The rats in the model group had interstitial infiltration of inflammatory cells, alveolar structure destruction, alveolar septum thicken, extensive alveolar hemorrhage, telangiectasia; the levels of TNF-α, IL-1β, IL-6, MDA and NF-κB p65 protein expression in lung tissue were significantly higher than those in the normal control group and sham operation group [TNF-α (ng/L): 132.81±16.15 vs. 45.08±5.98, 46.10±6.72, IL-1β (ng/L): 137.32±15.22 vs. 51.03±7.89, 50.92±8.13; IL-6 (ng/L): 138.39±14.28 vs. 51.68±7.03, 52.48±7.36; MDA (kU/g): 1.79±0.13 vs. 0.96±0.05, 0.97±0.05; NF-κB p65 protein (NF-κB p65/GAPDH): 2.82±0.23 vs. 1.76±0.12, 1.82±0.13; all P < 0.05], the activity of SOD decreased significantly (kU/g: 45.90±5.46 vs. 92.11±10.13, 93.36±10.56, both P < 0.05). The changes in lung histopathology in the forsythiaside A intervention group were obviously improved as compared with the model group, which showed less inflammatory cell infiltration, less alveolar septum thickening, less bleeding and more intact structures; the levels of TNF-α, IL-1β, IL-6, MDA and the expression of NF-κB p65 protein in lung tissue were significantly lower than those in the model group [TNF-α (ng/L): 72.48±9.78 vs. 132.81±16.15, IL-1β (ng/L): 83.85±12.46 vs. 137.32±15.22, IL-6 (ng/L): 81.88±11.89 vs. 138.39±14.28, MDA (kU/L): 1.29±0.09 vs. 1.79±0.13, NF-κB p65 protein (NF-κB p65/GAPDH): 2.29±0.19 vs. 2.82±0.23, all P < 0.05], SOD activity increased significantly (kU/g: 66.03±7.98 vs. 45.90±5.46, P < 0.05). Conclusions:Forsythiaside A can effectively alleviate ALI in septic rats. The mechanism may be related to down-regulate the expression of NF-κB p65 and reduce the level of inflammatory factors and free radicals in lung tissue, thereby against acute lung injury in septic rats.
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Objective:To evaluate the therapeutic effect of electroacupuncture on acute gastrointestinal injury (AGI) in patients with severe traumatic brain injury (sTBI).Methods:A prospective randomized controlled trial was conducted. 126 consecutively hospitalized patients with AGI after sTBI admitted to intensive care unit (ICU) of the First Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine from January 2018 to December 2019 were enrolled. The patients were divided into observation group and control group by random number table. All the patients of two groups were given conventional treatment of western medicine for consecutive 7 days, including the treatments of primary diseases, indwelling nasogastric tube to extract gastric contents every 6 hours to determine gastric residual volume (GRV). When vital signs were basically stable, enteral nutrition (EN) was implemented and EN feeding amount and speed were adjusted according to GRV. On the basis of conventional western medicine treatment, the observation group was treated with electroacupuncture at Zusanli, Tianshu, Shangjuxu, Xiajuxu and Zhongwan, once in the morning and once in the evening, 30 minutes each time. The gastrointestinal function parameters including intra-abdominal pressure (IAP), serum diamine oxidase (DAO) and gastrointestinal failure (GIF) scores were observed before treatment and at day 3 and day 7 of treatment. The incidence of ICU hospital-acquired pneumonia (HAP-ICU), duration of mechanical ventilation (MV), length of ICU stay, 28-day mortality and adverse reactions of electroacupuncture were also observed in the two groups. Kaplan-Meier method was used for 28-day survival analysis.Results:During the 7-day treatment and observation, 26 cases of 126 patients withdrew from the study, and 100 cases were actually enrolled, 50 cases in the observation group and 50 cases in the control group. IAP and DAO at day 3 of treatment in both groups were significantly lower than those before treatment [control group: IAP (cmH 2O, 1 cmH 2O = 0.098 kPa) was 13.75±2.76 vs. 18.11±3.97, DAO (U/L) was 129.88±24.81 vs. 158.01±22.64; observation group: IAP (cmH 2O) was 13.56±2.19 vs. 18.50±3.54, DAO (U/L) was 129.11±29.32 vs. 159.36±28.65; all P < 0.01]. The gastrointestinal function parameters of the two groups improved gradually with the extension of treatment time, and the IAP, DAO and GIF scores at day 7 of treatment in the observation group were significantly lower than those in the control group [IAP (cmH 2O): 11.28±3.61 vs. 12.68±3.23, DAO (U/L): 49.69±17.56 vs. 57.27±20.15, GIF score: 2.02±0.74 vs. 2.40±0.70, all P < 0.05). The duration of MV and the length of ICU stay in the observation group were significantly shorter than those in the control group [duration of MV (days): 15.72±4.60 vs. 18.08±4.54, length of ICU stay (days): 16.76±4.68 vs. 19.26±5.42, both P < 0.05], and the incidence of ICU-HAP and 28-day mortality were significantly lowered (12.0% vs. 30.0%, 22.0% vs. 32.0%, both P < 0.05). Survival analysis showed that the 28-day cumulative survival rate in the observation group was significantly higher than that in the control group (86.4% vs. 76.1%; Log-Rank test: χ 2 = 37.954, P < 0.001). The patients in the observation group had no significant adverse reaction of electroacupuncture treatment. Conclusion:Electroacupuncture at corresponding acupoints can effectively improve gastrointestinal function in patients with AGI after sTBI, which is beneficial to shortening the length of ICU stay, promoting the recovery of the patients, and reducing the 28-day mortality.
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Objective To discuss the influence of Tanshinone ⅡA on the tight junction protein of intestinal mucosal epithelial cells in rat severe septic models. Methods Seventy-five Sprague-Dawley (SD) rats were randomly divided into sham operation group, model group and Tanshinone ⅡA injection high (20 mg/kg), medium (10 mg/kg) and low (5 mg/kg) dose groups, each group 15 rats. Sepsis rat models were established by cecal ligation and puncture (CLP) method, in sham operation group, only switched abdominal surgery was performed without CLP. In Tanshinone ⅡA injection groups, different doses of Tanshinone ⅡA were injected intraperitoneally after modeling for 10 minutes and 6 hours; in sham operation and model groups, equal volume of normal saline was injected intraperitoneally at the same times as above. After operation, 3 L/kg of normal saline was injected into the caudal vein in all rats for fluid resuscitation.Twelve hours after operation, the rats were killed, the abdominal lymph nodes, liver, spleen and kidney tissues were taken for bacterial culture and calculating the rate of bacterial translocation; under microscope, the histopathological changes of ileum mucosal tissues were examined and Chiu scoring was carried out; TdT-mediated dUTP nick end labeling (TUNEL) was applied to detect the ileum mucosal epithelial cell apoptosis and calculating the index (AI);fluorescence immunoassay and Western Blot methods were used to measure the contents and protein expression levels of tight junction protein, junctional adhesion molecule-1 (JAM), Claudin-1, Zonula occludens-1 (ZO-1), Occludin, c-Fos and Tryptase. Results ① In bacterial cultures of abdominal lymph node, liver, spleen and kidney, the positive rate of mesenteric lymph node was the highest, followed by liver and spleen, mainly Escherichia coli, Proteus mirabilis, etc. The highest positive rate of bacterial culture was in model group (38.8%), followed by low dose of Tanshinone ⅡA injection group (35.0%), and the lowest was 16.6% in high dose Tanshinone ⅡA injection group, the differences being statistically significant in comparisons between any pair of groups (all P < 0.05). ② Pathological examination showed that the pathological changes of ileum mucosa were obvious and the Chiu score (4.17±0.98 vs. 0) and AI (11.70±2.87 vs. 2.17±0.80) in model group were significantly higher than those in sham group (all P < 0.05); with the increase of dosage of Tanshinone ⅡA injection, the pathological changes of rat ileum mucosa were improved gradually, the Chiu score and AI were decreased gradually, and the degrees of decrease in high dose Tanshinone ⅡA group were more significant than those in model group (Chiu score: 1.12±0.79 vs. 4.17±0.98, AI: 3.65±1.98 vs. 11.70±2.87, both P < 0.05).③ Immunofluorescence staining showed that the positive staining of protein JAM, ZO-1 and c-Fos were all green in color, Claudin-1, Occludin and Tryptase were all red in color, the localizations of all of them were in the cytoplasm, the protein expression of JAM, Claudin-1, ZO-1, Occludin from strong to weak in turn were Sham group, high, medium, low dose Tanshinone ⅡA group and model group, the expression of c-Fos, Tryptase from strong to weak in turn were model group, low, medium, high dose Tanshinone ⅡA group and Sham group. ④ Western Blot showed that the expressions of ileum tissue JAM, Claudin-1, ZO-1 and Occludin in model group were all significantly lower than those of the sham group, while the expressions of c-Fos, Tryptase were obviously higher than those of the sham group, with the increase of dosage of Tanshinone ⅡA, the expressions of JAM, Claudin-1, ZO-1 and Occludin were increased gradually and the protein expressions of c-Fos and Tryptase were gradually decreased, and the changes in high dosage group of Tanshinone ⅡA were more significant than those in low and moderate groups [JAM (gray value): 25.39±1.82 vs. 12.41±1.34, 19.45±1.66, Claudin-1 (gray value): 28.44±1.56 vs.17.26±1.46, 21.23±1.34, ZO-1 (gray value): 28.84±1.59 vs. 16.45±1.21, 24.22±1.46, Occludin (gray value): 25.49±1.63 vs. 13.34±1.45, 19.45±1.37, c-Fos (gray value):15.76±1.36 vs. 27.84±1.36, 21.22±1.73, Tryptase (gray value): 14.44±1.41 vs. 28.14±1.38, 22.32±1.57], all the above comparisons of different dosage groups were statistically significant (all P < 0.05). Conclusion Tanshinone ⅡA injection may improve intestinal wall structure and reduce bacterial translocation by improving the intestinal mucosal tight junction protein in sepsis model rats, and this effect is positively correlated to Tanshinone ⅡA dosage.
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The benefits of early enteral nutrition (EEN) during critical illness have been widely accepted by global experts. To popularize this new concept and provide standardized, reasonable and effective EEN therapy for critically ill patients in China, more than 20 experts from throughout the country discussed and developed this consensus. We used the GRADE approach for consensus development, focusing on important clinical issues such as nutrition assessment, initiating mode, route selection and tolerance monitoring of EEN support therapy for current critically ill patients. This consensus would be certainly help for intensive care physicians in the clinical application of EEN support therapy for critically ill patients.
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Humanos , China , Consenso , Cuidados Críticos , Estado Terminal , Nutrição EnteralRESUMO
Objective To develop a traditional Chinese medicine (TCM) syndrome score scale for acute gastrointestinal injury (AGI) in sepsis, and to carry out its reliability and validity analyses and its clinical preliminary application. Methods ① According to the characteristics of intensive care unit (ICU) patients, combined with the understanding of etiology, pathogenesis and physical signs of TCM and literature search, a preliminary framework of scoring system for TCM syndromes of AGI in sepsis was constructed to carry out the scoring by this scale. ② After the scale and data were obtained, the analyses of split-half reliability (indicated by Guttman's split-half reliability of the a and b groups), test-retest reliability and the internal consistency reliability (expressed by the Cronbach's coefficient α) were carried out, and the structural validity and criterion validity were also analyzed. ③ The AGI patients were divided into two groups according to the 28-day survival and death conditions, and the AGI TCM syndrome score, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score, sequential organ failure assessment (SOFA) score, and multiple organ dysfunction syndrome (MODS) score were compared between the two groups to determine the best cut-off point for survival analysis. Results ① The first draft of the septic AGI TCM syndrome rating scale was prepared, The TCM syndrome indicators include: abdominal distension, constipation/diarrhea, diet situation, vomiting/stomach retention, tongue proper, tongue coating, pulse manifestation, belching, body temperature, and accompanied syndrome, there were 6 points for scoring, 0 - 6 points, and they were divided into normal (0 points), mild (2 points), moderate (4 points), and severe (6 points) in severity. ② Eighty-eight patients with septic AGI were included in the final statistics. The retest of correlation coefficient of this scale was R = 0.974 (> 0.85), Guttman's split-half reliability was 0.793 (> 0.7) and the Cronbach's coefficient α was > 0.7. This scale was suitable for factor analysis. After rotation, 3 factors were determined, which were named as TCM syndrome differentiation, related physical signs, and gastrointestinal tolerance. After modeling, the confirmatory factor analysis showed that the model approximate error root mean square (RMSEA) was 0.07 (< 0.08), and the goodness of fit index (CFI) = 0.90; the Pearson correlation analyses between the criteria validity of APACHE Ⅱ, SOFA, MODS scores and TCM 1 score and TCM 2 score of this scale showed that the r values were 0.802 and 0.752, 0.524 and 0.519, 0.619 and 0.590, respectively, all P < 0.01. ③ Compared with the survival group, TCM score (33.73±5.95 vs. 37.28±5.26, t = 2.945, P = 0.004), the APACHE Ⅱ score (19.90±4.47 vs. 22.28±5.79, t = 2.069, P = 0.043), SOFA score (8.73±1.11 vs. 9.64±1.38, t = 3.329, P = 0.020) in the death group were significantly decreased; MODS score in the death group showed a decreasing trend (6.65±1.22 vs. 7.28±1.60, t = 2.078, P = 0.050). Cox regression analysis showed that when the survival analysis was performed with a cut-off point of 35, the 28-day survival rate of patients with TCM syndrome score ≥ 35 was significantly lower than that of patients with < 35 score, χ2= 6.362, P = 0.012. Conclusions The TCM syndrome rating scale for AGI in sepsis was successfully prepared. The statistical reliability and validity of this scale are good. Preliminary clinical application shows that this scale can predict the prognosis and severity of patients with septic AGI. Trial registration China Clinical Trial Registry Center, ChiCTR-IOR-15007625.
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Objective To investigate the prevalence of feeding intolerance (FI),and to explore the FI within 7 days of ICU admission in association with clinical outcome in critically ill patients.Methods The adult patients from 14 general ICUs in Zhejiang Province with an expected admission to ICU for at least 24h were recruited from March 2014 to August 2014,and all clinical,laboratory,and survival data were prospectively collected.The AGI (acute gastrointestinal injury) grade was daily assessed based on gastrointestinal (GI) symptoms,feeding details and organ dysfunction within the first week of ICU stay.The intra-abdominal pressures (IAP) was measured using AbViser device.Results Of 550 patients enrolled,418 were assessed in GI symptoms and feeding details within 7 days of ICU stay.The mean age and SOFA score were (65.1 ± 18.3) years and (8.96 ±4.10),respectively.Of them,355 patients (84.9％) were under mechanical ventilation support,and 37 (8.85％) received renal replacement therapy.The mean length of time for enteral feeding was (30.8 ±26.2) h,and the prevalence of FI on the 3rd and 7th day of ICU stay accounted for 39.2％ and 25.4％,respectively.Compared to those with FI within 7 days of ICU stay,the patients without FI had higher rate of successively weaning from mechanical ventilation (21.3％ vs.5.7％,P =0.003) and higher rate of withdrawal of vasoactive medication (45.5％ vs.20.0％,P =0.037),as well as lower mortality rate of 28-day (24.4％ vs.38.7％,P =0.004) and 60-day (29.6％ vs.44.3％,P =0.005).In multivariate Cox regression model with adjustment for age,sex,participant center,serum creatinine and lactate,AGI grade on the first day of ICU stay,and comorbidities,the FI within 7 days of ICU stay (x2 ≥ 7.24,P ＜ 0.01) remained to be independent predictors for 60-day mortality.After further adjusted for SOFA score,the FI within 7 days of ICU stay (HR =1.71,95％ CI:1.18-2.49;P =0.006) and AGI grade on the first day of ICU stay (HR =1.33,95 ％ CI:1.07-1.65;P =0.009) could provide independent prognostic values of 60-day mortality.Conclusions There is high rate of FI occurred within 7 days of ICU stay,and is significantly associated with worse outcome.In addition,this study also provides evidence to further support that measurement of gastrointestinal dysfunction could increase value of SOFA score in outcome prediction for the risk of 60-day mortality.
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[Objective] To further study tanshinone role in intestinal barrier dysfunction in sepsis. [Methods] Mainly refer to nearly a decade at home and abroad on tanshinone pharmacological effects, and tanshinone in sepsis, especially the impact on the intestinal barrier dysfunction. [Results] Tanshinone in sepsis, and heart head blood-vessel, anti-tumor and so on in the field of study is more, but improving sepsis of intestinal barrier function in aspects of the study is less, but through tanshinone antibacterial anti-inflammatory, antioxidant, anti platelet aggregation, improve microcirculation, regulate immune function, this paper expounds tanshinone improving sepsis, possible mechanisms of intestinal barrier function. [Conclusion] Tanshinone can improve sepsis pathophysiological process, ischemia oxygen to the tissues and organs and whole body strong inflammatory reaction factors such as the cause of intestinal barrier function is impaired, but for its specific mechanism, further research is needed.
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Objective To observe the effect of Shenfu injection on intestinal function in rats with sepsis. Methods Forty Sprague-Dawley (SD) rats were randomly divided into four groups: sham operation, sepsis model, low and high concentration Shenfu injection groups, each groupn = 10. The sepsis model was replicated by cecal ligation and puncture (CLP), while the rate in sham operation group just underwent abdominal incision without CLP. Ten minutes after CLP, the low and high dose Shenfu injection groups were given 5 mL/kg and 10 mL/kg Shenfu intravenous injection via a tail vein respectively. The rats in the model group were treated by intravenous injection of 10 mL/kg normal saline through a tail vein in 10 minutes after CLP. Twelve hours later, the rats were sacrificed. The levels of Ghrelin, Gastrin, tumor necrosis factor-α (TNF-α), high mobility group B1 protein (HMGB1), myeloperoxidase (MPO) and diamine oxidase (DAO) activity in serum were detected by enzyme linked immunosorbent assay (ELISA). The levels of protein of Ghrelin and gastrin receptor (GHSR) were detected by Western Blot. Under light microscope, the histopathological changes in intestinal mucosa were investigated, and Chiu score was determined, and the apoptosis index (AI) of intestinal mucosal epithelial cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).Results Compared with sham operation group, in model group, the levels of Ghrelin and Gastrin in serum were significantly decreased [Ghrelin (ng/L): 121.23±3.53 vs. 146.28±5.43, Gastrin (ng/L): 81.78±3.27 vs 102.78±4.07], the serum levels of TNF-α and HMGB1 were markedly increased [TNF-α (mg/L): 93.71±3.66 vs. 11.69±1.44, HMGB1 (mg/L): 76.25±4.12 vs. 22.41±3.08], the DAO activity and protein expressions of Ghrelin and GHSR of intestinal tissue were obviously decreased [DAO (U/mL): 14.64 ±0.68 vs. 25.13±1.98, Ghrelin (grey value): 0.12±0.02 vs. 0.23±0.04, GHSR (grey value): 0.18±0.02 vs. 0.32±0.03], the MPO activity in intestinal tissue, Chiu score of intestinal mucosa and AI of ileum mucosal epithelial cells were remarkably increased [MPO (mg/L): 175.98±6.95 vs. 45.64±4.48, Chiu score: 3.90±0.52 vs. 0.30±0.30, AI: 29.31±1.65 vs. 5.45±1.35, allP 0.05). Under light microscope, the pathological changes were seen as follows: destruction and obvious edema of intestinal mucosal villi, ulcer formation, significant perivascular hemorrhage, presence of neutrophil infiltration and fracture of basement membrane in model group, while in low and high Shenfu groups, the intestinal villi had little defect, focal necrosis, small amounts of hemorrhage and neutrophil infiltration. Conclusions Shenfu injection can significantly improve the abnormal expressions of serum Ghrelin, reduce the levels of serum TNF-α and HMGB1, lowered MPO activity and enhance DAO activity in intestinal tissue, alleviate pathological changes in ileum mucosa, and decrease AI of ileum mucosal epithelial cells in rats with sepsis. And the degree of therapeutic effect is proportional to the Shenfu injection dose.
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Objective] To investigate the protective effect and potential mechanism of Astragalus injection on mucosa epithelial cells in rats with sepsis . [Methods] Thirty Sprague-Dawley rats were randomly divided into sham,model and Astragalus injection treatment groups with 10 rats in each group.Rat sepsis model was constructed by cecal ligation and puncture.Rats in the Astragalus injection treatment group received 800 mg·kg-1 of Astragalus injection at 15 minutes before and 6 hours after operation. Intestinal mucosa were harvested and frozen at -80℃ on post-operation 24 hours. Intestinal mucosa epithelial cell apoptosis was detected by TUNEL,while Bcl-2 and Bax levels were determined by real-time quantitative PCR(qPCR) and Western blot technique.[Results] The apoptosis indexes were 5.2±0.3,22.3±1.2 and 11.2±0.9 of sham,model and Astragalus injection treatment group,respectively. Apoptosis index in model group was significantly higher than that in sham group( P<0.05),while the apoptosis of mucosa epithelial cells was significantly attenuated after Astragalus injection treatment(P<0.05).In model group,the expression of Bcl-2 was lower than that of sham group,whereas the expression of Bax was higher than that in sham group(P<0.05).Astragalus injection treatment significantly lowered Bax level while elevated Bcl-2 expression, compared with model group(P<0.05). [Conclusion] Astragalus injection could suppress sepsis-induced apoptosis of mucosa epithelial cells via up-regulating Bcl-2 and down-regulating Bax.
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Objective To study the effects of pneumonia caused by Klebsiella pneumoniae on apoptosis of thymocytes in rats and its possible mechanism. Methods A total of 48 Sprague Dawley rats were randomly (random number) divided into 2 groups, namely the control group (n =24) and the infection group ( n = 24). The pneumonia models of rats were made with 0.3 mL Klebsiella pneumoniae suspension administered intratracheally per animal. On the 2nd, 4th, and 6th day after intratracheal instillation of bacteria, 1/ 3 of the rats in each group were sacrificed and TdT-mediated dUTP nick end labeling (TUN EL) method was used to assess the apoptosis of thymocytes. The expressions of cleaved Caspase-3, Bcl-2 and Fas in thympcytes of rats were detected with immunohistochemical staining. Results On each interval, apoptosis index of thymocytes, and the expressions of Cleaved Caspase-3 and Fas in the infection group were all higher than those in the control group (P < 0.01) , while the expressions of Bcl-2 lower than those in the control group (P<0.01). As more time consumed, the apoptotic index of thymocytes and the expressions of cleaved Caspase-3 in the infection group increased significantly (P<0.05). The expressions of Bcl-2 declined gradually (P < 0.05), but the expressions of Fas reached their peak 4th day after infection. There were no significant dynamic changes in all above mentioned variables in control group. Conclusions Pneumonia caused by Klebsiella Pneumoniae can lead to the increase in thymocyte apoptosis in rats. The mechanism may be associated with the decreased expression of Bcl-2 and the increased expression of Fas in thymocytes of rats with pneumonia caused by Klebsiella pneumoniae. The different apoptosis regulation pathways have different effects on different phase of pneumonia, that the effects of Fas decrease 4th day after pneumonia, while the effects of Bcl-2 increase further.
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Objective To study the effect of anti-IL-5 monoclony antibody (TRFK-5) on migration of Eos from BM to the airways in sensitized mice. Methods Male C57BL/6 (6-8wk of age) murine model of Asthma and control group were estabolished with routine method. The outcome measurements include white blood cell (WBC) total count, differential count of bronchoalveolar lavage fluid (BALF) and peripheral blood (PB), nuclear cell count and eosinophil percentage of BM. These parameters were collected 12 h after the final allergen challenge. To cheek Eos infiltration, the histology of lung tissues was also observed. Further, the effects of intranasal TRFK-5 on above changes were investigated. Results Eosinophil numbers of BALF, PB, BM and the infiltration of Eos in pulmovnary tissues were increased considerably 12 h after final OVA challenge compared with negative group(P
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0.05).The amount of insulin in ZL group was significantly lower than that in the control group(P