Effect of Guizhitang on Atherosclerosis by Regulating Immune and Intestinal Flora / 中国实验方剂学杂志

Objective:To observe the effect of Guizhitang （GZT） on peripheral blood monocytes， intestinal flora and AS plaque formation of ApoE^-/- mice induced by Western diet （WD）. Method:In this study， 40 12-week-old homozygous female ApoE^-/- mice were randomly divided into chow diet （CD） group （ApoE^-/-+CD）， WD group （ApoE^-/-+WD）， GZT group （ApoE^-/-+WD+GZT， 7.83 g·kg^-1） and atorvastatin （Atr） group （ApoE^-/-+WD+Atr， 3.33 mg·kg^-1）. And 10 matched C57BL/6 mice were set as wild CD control group （C57BL/6+CD）. Except the CD group， the rest groups were given WD to induce models. Treatment groups were given Guizhitang or atorvastatin orally in addition to WD， while ApoE^-/-+CD and ApoE^-/-+WD model groups were treated with the same volume of double steam water at the same time. After 4 weeks of intervention， 5 mice in each group were selected to collect the eyeball blood samples. The levels of plasma lipids were detected by automatic biochemical analyzer， and the proportion of peripheral blood mononuclear cells and its subtypes， and the expression levels of surface receptors toll like receptor 4 （TLR4） and CD36 were detected by flow cytometry， the intestinal flora of mice was detected by 16S rDNA sequencing. The remaining 5 mice in each group were intervened for 12 weeks， and the aorta was taken to detect the formation of aortic plaque by oil red O staining. Result:After intervention for 4 week， compared with C57BL/6+CD group， the levels of plasma total cholesterol （TC） and low-density lipoprotein （LDL） levels in ApoE^-/-+CD and ApoE^-/-+WD groups were increased （<italic>P</italic><0.01）. ApoE^-/-+WD group showed increase in the proportion of monocytes， their inflammatory subtypes Ly6C^__， and TLR4 expression on monocyte surface in blood （<italic>P</italic><0.05）. ApoE^-/-+WD group induced the imbalance of intestinal flora， with increase of Firmicutes and decrease of Verrucomicrobia in ileum of ApoE^-/-mice. Compared with ApoE^-/-+WD group， there was no significant change in blood lipid level and monocyte proportion in ApoE^-/-+WD+GZT group， but with decrease in the proportion of Ly6C^__， increase in the proportion of anti-inflammatory subtype Ly6C^-， and decrease in the expression of TLR4 and CD36 on monocyte surface （P<0.05）. ApoE^-/-+WD+GZT group showed decrease of Firmicutes and increase of Bacteroidetes and Verrucomicrobia in ileum of ApoE^-/-mice. After 12 weeks of intervention， ApoE^-/-+WD group showed increase in the number and area of aortic plaques in ApoE^-/-mice. ApoE^-/-+WD+GZT group showed decrease of the area of aortic AS plaques. Conclusion:GZT can reduce the immune damage and imbalance of intestinal flora caused by WD， then inhibit the formation of AS plaque.

Effect of Qingzao Jiufei decoction and its decomposing agent on lung inflammation?related factors in mice infected with Mycoplasma pneumoniae / 中国实验动物学报

Objective To investigate the effects of Qingzao Jiufei Tang(QJD)and its decomposing agent on the levels of TNF-α,INF-γ levels of TNF-α of Qingo, P1 and AQP5 in lung tissue of mice infected with Mycoplasma pneu-moniae(MP),and to clarify its molecular anti-infective mechanism. Methods One hundred-forty-four SPF grade BABL/c mice were randomly divided into the normal group(group A),model group(group B),QJD group(group C),QJD de-composition agent I group(group D)、QJD decomposition group II Group(group E)and azithromycin group(group F), with 24 rats in each group. Besides the group A,the other 5 groups of mice were treated with MP infection. After the mod-eling,the mice were given corresponding drugs by gastric gavage,and samples were obtained on the days 3,7,10,14 af-ter the model was established. The lung tissue sections were examined by histopathology, and the degrees of inflammation in lung tissues in the mice were evaluated,and the lung index and the ratio of dry and wet lung weight ratio in the mice were calculated. The levels of MPN372 and P1 genes were determined by qPCR assay. The serum TNF-α and INF-γ levels in the mice were assessed by enzyme linked immunosorbent assay(ELISA). The expression of AQP5 protein was detected by Westernblot. Results After MP infection, the pathological examination revealed thickening of alveolar septum and bronchioles,and extensive inflammatory cell infiltration in the lung tissues. The lung index was increased and the ratio of dry and wet lung weight(P<0.05). TNF- α and INF-γ cedd and was increased, and reached the peak on the seventh day. The expression of AQP5 protein showed a downward tendency,and began to gradually increase on the 14th day. Com-pared with the group B,the expression levels of MPN372,P1 and TNF-α in the group D were down-regulated,and the ex-pression levels of INF-γ levein the group E were up-regulated from the 7th day. Conclusions QJD can control pulmonary inflammation in mice after MP infection. The decrease of production of MP toxin MPN372 and the expression of adhesion protein P1,the up-regulation of expression of INF-γ and AQP5 proteins,and down-regulation of TNF-α expression are one of the mechanisms of its action.

Effect of cathepsin L activation on trichloroethylene-induced skin allergy in mice / 中国职业医学

OBJECTIVE: To observe the expression of cathepsin L( CTSL) on the skin of trichloroethylene( TCE)-sensitized mice,and explore the mechanism of CTSL in TCE-induced immunological skin damage. METHODS: The specific pathogen free female BALB/c mice were randomly divided into blank control group( n = 5),solvent control group( n = 5),TCE group( n = 15) and inhibitor group( n = 15). Skin sensitization experiments were performed according to the maximum guinea pig test method. The TCE group and inhibitor group were divided into sensitized subgroups and non-sensitized subgroups according to skin sensitization results. The skin tissues were taken 72 hours after the last TCE challenge.Hematoxylin-eosin staining was used to observe the pathological structure of skin tissues and measured the thickness of epidermis. The level of Ctsl mRNA was examined by real-time quantitative polymerase chain reaction,and the expression of CTSL, interleukin( IL)-6 and IL-17 were studied by immunohistochemical staining technique. RESULTS: The sensitization rate of TCE group and inhibitor group were 40. 0%(6/15) and 33. 3%(5/15) respectively. There was no statistical difference in the sensitization rate between the two groups( P > 0. 05). The thickness of epidermis and relative expression of Ctsl mRNA,CTSL,IL-6 and IL-17 in TCE sensitized subgroup and inhibitor sensitized subgroup were higher than that in blank control group,solvent control group,TCE non-sensitized subgroup and inhibitor non-sensitized subgroup(P < 0. 05). The above-mentioned indexes were higher in TCE sensitized subgroup than that in inhibitor sensitized subgroup( P < 0. 05). The relative expression of Ctsl mRNA,CTSL,IL-6 and IL-17 in skin of TCE sensitized subgroup were positively correlated between any two indexes( P < 0. 05). CONCLUSION: CTSL activation may play an important role in immunological skin damage of TCE-sensitized mice,which may be related to the promotion of IL-6 and IL-17 release.

Diagnostic value of serum procalcitonin for infection in the immunocompromised critically ill patients with ;suspected infection / 中华危重病急救医学

Objective To evaluate the diagnostic and prognostic value of the serum procalcitonin ( PCT ) level in the non-acquired immune deficiency syndrome ( AIDS ) immunocompromised critically ill patients suspected to have infection. Methods A retrospective study was conducted in the non-AIDS immunocompromised patients who were admitted to Department of Critical Care Medicine of Xiangya Hospital, Central South University during January 2011 to December 2014. Demographic characteristics, underlying disease, acute physiology and chronic health evaluationⅡ( APACHEⅡ) score at admission, and clinical records including baseline and peak levels of temperature, white blood count ( WBC ), PCT, and survival rate within 28 days, infection focus, infectious agents ( bacterial, fungi or mixed infection ), and the severity of infection ( sepsis, severe sepsis, or septic shock ) were recorded. Receiver operating characteristic ( ROC ) curve was plotted, and the diagnostic and protective value of above parameters was evaluated. Results A total of 98 patients ( 43 male and 55 female ) were enrolled in the study with a median age of 44 ( 28, 52 ) years old and a median APACHEⅡscore of 17 ( 11, 20 );47 with malignant hematological tumor, 45 with autoimmune diseases, and 6 post solid organ transplantation. Among them 53 patients ( 54.1%) died within 28 days. Twenty-seven patients were diagnosed as systemic inflammatory response syndrome ( SIRS ) without infection. Among 71 patients with infection, 45 were diagnosed as bacterial infection, 10 with fungal infection, and 16 with mixed infection. Sepsis was diagnosed in 7 patients, severe sepsis in 32 patients , and septic shock in 32 patients .①There was no statistical significance in the baseline and peak levels of PCT and WBC, or baseline level of temperature between the groups of SIRS patients without infection and infected patients. The peak level of temperature was significantly higher in the patients with infection as compared with that of the SIRS without infection patients [℃:39.4 ( 38.9, 40.0 ) vs. 38.8 ( 37.8, 39.2 ), Z=-3.268, P=0.001 ]. It was showed by subgroup analysis that in patients with hematological malignant disease or autoimmune diseases, higher level of body temperature was found in infection group compared with non-infection SIRS group [℃:39.5 ( 39.0, 40.0 ) vs. 39.0 ( 38.4, 39.4 ), Z=-2.349, P=0.019;39.0 ( 38.4, 39.5 ) vs. 38.2 ( 37.0, 38.9 ), Z=-2.221, P=0.026 ].②The baseline level of PCT (μg/L ) were 0.54 ( 0.20, 4.19 ), 2.78 ( 0.50, 9.54 ), 1.00 ( 0.45, 6.89 ), and 0.22 ( 0.07, 1.86 ) in non-infection SIRS patients or the patients with bacterial, fungal, and mixed infection, respectively. The peak level of PCT (μg/L ) were 4.19 ( 1.95, 13.42 ), 12.37 ( 3.82, 45.89 ), 1.82 ( 0.49, 17.86 ), and 5.14 ( 2.66, 12.62 ), respectively, in each subgroup. When the comparison was conducted among the patients with different infectious agent, the baseline level of PCT in patients with bacterial infection was significantly higher than that in SIRS patients without infection ( P=0.026 ) and mixed infection patients ( P=0.001 ), and the peak level of PCT was significantly higher than that in the SIRS patients without infection ( P=0.009 ) and the patients with fungal infection ( P=0.016 ). ROC curve showed that the higher value was found in the baseline and peak levels of PCT for diagnosis of septic shock in all patients [ area under ROC curve ( AUC ) of baseline level = 0.681±0.054, P = 0.001; AUC of peak level = 0.690±0.054, P=0.002 ], and the same value was also found in the baseline and peak levels of PCT for diagnosis of bacterial infection in the patients with malignant hematological tumor ( AUC of baseline level=0.687±0.080, P=0.008;AUC of peak level=0.697±0.079, P=0.021 ).③The peak level of PCT (μg/L ) were 4.05 ( 0.53, 31.22 ), 5.78 ( 2.14, 16.68 ), and 11.64 ( 2.94, 58.14 ) in subgroup of patients with sepsis, severe sepsis and septic shock, respectively, and they showed no statistical significance among subgroups ( P>0.05 ). A high serum level of peak PCT strongly indicated the presence of septic shock ( AUC=0.646±0.060, P=0.019 ), especially in the subgroup of patients with systemic autoimmune disease ( AUC=0.689±0.081, P=0.035 ).④The peak level of PCT (μg/L ) in the APACHEⅡ>18 group ( 38 cases ) was significantly higher than that of APACHEⅡ≤18 group [ 60 cases, PCT (μg/L ):11.64 ( 3.36, 39.39 ) vs. 4.42 ( 1.32, 14.70 ), P=0.016 ];there was a certain correlation between the peak level of PCT and the severity of the disease.⑤The peak level of PCT in death group was significantly higher than that of the survival group [μg/L:9.07 ( 3.05, 33.09 ) vs. 4.19 ( 1.26, 14.61 ), P=0.043 ]. ROC curve showed that the peak level of PCT might be valuable in predicting the prognosis in immunocompromised patients ( AUC=0.619±0.057, P=0.043 ). Conclusions The serum level of PCT is found to be a reliable marker for the diagnosis of bacterial infection in immunocompromised critical patients, especially in those with hematologic malignancy. Additionally, PCT provides a useful tool for evaluating the severity of infection and the prognosis of critically ill patients.