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.

Objective To investigate the risk factors of prognosis of gram-positive bacteria associated nosocomial bloodstream infections,and to investigate the drug resistance of the strains.Methods A total of 132 patients with gram-positive bacteria associated nosocomial bloodstream infections were collected from the First Affiliated Hospital of Zhejiang Chinese Medicine University during January 2010 and December 2012.Clinical data including demographic characteristics,underlying diseases,risk factors and use of antibacterial agents were retrospectively analyzed.According to 28-day prognosis,patients were divided into survival group (n =97) and death group (n =35).Binary logistic regression was used to identify the risk factors of 28-day fatality.Results Among 132 patients,49 (37.12％) were infected with coagulase-negative Staphylococcus,46 (34.85％) were infected with Staphylococcus aureus,37 (28.03％)were infected with Enterococcus.The rates of methicillin resistant coagulase negative Staphylococci (MRCNS) and methicillin-resistant Staphylococcus aureus (MRSA) were 77.55％ (38/49) and 54.35％ (25/46),respectively.The rate of linezolid resistant coagulase negative Staphylococci was 8.16％ (4/49) ; Four out of 37 strains (10.81％) of Enterococcus were both resistant to vancomycin and linezolid.Binary logistic regression showed that septic shock (OR =34.344,95％ CI:6.539-180.389,P =0.000),deep venous catheterization (OR =13.411,95％ CI:1.877-95.832,P =0.010),no catheter removal after infection (OR =8.759,95％ CI:2.197-34.911,P =0.002),parenteral nutrition (OR =3.684,95％ CI:1.072-12.663,P =0.038),inappropriate antibacterial therapy in early stage (OR =12.951,95％ CI:2.075-80.836,P =0.006) and Enterococcus associated bloodstream infections (OR =4.227,95％ CI:1.090-16.394,P =0.037) were independent risk factors of 28-day fatality in patients with gram-positive bacteria associated nosocomial bloodstream infections.Conclusions The predominant pathogens are coagulase-negative Staphylococcus,Staphylococcus aureus and Enterococcus in gram-positive bacteria associated nosocomial bloodstream infections.Patients with septic shock,deep venous catheterization,no catheter removal after infection,parenteral nutrition,inappropriate antibacterial therapy in early stage and Enterococcus associated bloodstream infections are likely to have high fatality rate.

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.

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.