Curcumin inhibits NLRP3 inflammasome reduction on the early myocardial damage in septic rats / 中华急诊医学杂志

Objective:To investigate the effect of curcumin on the nucleotide-binding oligomerization domain-like receptor protein (NLRP3) inflammasome in cardiomyocytes of rats with early sepsis and its mechanism.Methods:Twenty-four male SD rats were randomly divided into three groups: the sham-operated group (sham group, n=8), the sepsis group ( n=8), and the curcumin intervention group (Cur group, n=8). A rat model of sepsis was prepared by cecal ligation and perforation (CLP). After modeling, 100 mg/kg of curcumin was intraperitoneally injected and repeated 24 h later. Rats in the sepsis group were injected with normal saline. The levels of myocardial injury-specific troponin T (cTnT) in rat plasma were detected by ELISA at 6, 24, and 48 h. Hematoxylin-eosin (HE) staining was used to observe the pathological injury of the myocardium in the myocardial tissue of rat at 48 h. The apoptosis of cardiomyocytes was detected by TUNEL assay. The expression levels of Cleaved caspase-1, NLRP3 and IL-1β protein were detected by Western blot, and the ultrastructural changes of the cardiomyocytes were observed under a transmission electron microscope. Results:The levels of cTnT in rat plasma at 6, 24, and 48 h in the Cur group were significantly lower than those in the sepsis group ( P<0.05). HE staining showed infiltration, cell edema and necrosis of myocardial inflammatory cells in the sepsis group, while only partial cell edema and necrosis were observed in the Cur group. TUNEL assay showed that the apoptosis of cardiomyocytes in the Cur group was significantly lower than that in the sepsis group [(28.4±2.3)% vs. (43.6±3.8)%, P<0.05]. The expression levels of Cleaved caspase-1, NLRP3 and IL-1β in the Cur group were lower than those in the sepsis group and higher than those in the sham-operated group ( P<0.05). Under transmission electron microscopy, the nuclei in the sham-operated group had intact membranes and uniform chromatin distribution; in the sepsis group chromatin margination, pyknosis, mitochondrial cristae breakage, cavitation, and partial breakage of sarcomere were observed; while in the Cur group partial chromatin margination, slightly edema and dilation of mitochondria, with basically complete morphology were observed. Conclusions:Curcumin inhibits NLRP3-mediated acute myocardial injury in septic rats, and its mechanism may be related to pyroptosis induced by the down-regulation of the expression of Cleaved caspase-1 and IL-1β protein.

Traditional Chinese Medicine excessive pattern correlation analysis of chronic obstructive pulmonary disease complicated acute exacerbation of chronic pulmonary heart disease / 国际中医中药杂志

Objective:To explore the correlation between Traditional Chinese Medicine (TCM) excessive patterns and clinical characteristics of acute exacerbation of chronic obstructive pulmonary disease (COPD) complicated with chronic pulmonary heart disease (CPHD) in high altitude environment.Methods:Patients with acute exacerbation of COPD complicated with CPHD admitted to the Pulmonology Department of Qinghai Provincial Hospital of Traditional Chinese Medicine from December 2016 to November 2017 were selected. Demographic data and clinical medical characteristics data of the patients were collected, and TCM patterns differentiation was conducted. The correlation between each pattern type and clinical characteristics and all collected laboratory indexes were analyzed by multivariate logistic regression.Results:Phlegm obstructing lung pattern showed a negative correlation relationship with mMRC score [ OR=0.419, 95% CI (0.219-0.802), P=0.009], PCT [ OR=8.132×10 -11, 95% CI (1.632×10 -16-4.1×10 -5), P<0.001], Hb [ OR=0.971, 95% CI (0.952-0.989), P=0.002] and PaCO 2[ OR=0.914, 95% CI (0.853-0.980), P=0.011]; turbid phlegm obstructing lung pattern showed a negative correlation relationship with gender(0 male, 1 female) [ OR=0.427, 95% CI (0.204-0.892), P=0.024], Hb [ OR=0.960, 95% CI (0.945-0.975), P<0.001], and there was a positive correlation relationship with LVEF [ OR=1.061, 95% CI (1.006-1.118), P=0.028]; phlegm-heat obstructing lung pattern showed a negative correlation relationship with Hb [ OR=0.950, 95% CI (0.927-0.974), P<0.001]and cardiac function grade [ OR=0.468, 95% CI (0.248,0.881), P=0.019], and there was a positive correlation relationship with PCT [ OR=1.118×10 8, 95% CI (1.466×10 4-8.523×10 11), P<0.001] and D-D [ OR=2.283, 95% CI (1.300-4.010), P=0.004]; there was a negative correlation between phlegm and stasis blocking lung pattern with cardiac function grade[ OR=0.309, 95% CI (0.167-0.570), P<0.001], and there was a positive correlation relationship with Hb[ OR=1.060, 95% CI (1.042-1.078), P<0.001]; there was a negative correlation between wet phlegm and blood stasis heat pattern with PCT [ OR=1.266×10 -13, 95% CI (1.658×10 -21-0.1×10 -4), P<0.001], SaO 2 [ OR=0.934, 95% CI (0.892-0.979), P=0.004], LVEF [ OR=0.896, 95% CI (0.826-0.971), P=0.008], D-D [ OR=0.030, 95% CI (0.002-0.508), P=0.015], and there was a positive correlation relationship with CRP [ OR=1.042, 95% CI (1.018-1.067), P<0.001], RBC [ OR=3.411, 95% CI (1.684-6.910), P<0.001], cardiac function grade [ OR=8.573, 95% CI (2.410-30.504), P<0.001], pulmonary arterial pressure difference [ OR=2.091, 95% CI (1.243-3.516), P=0.005]. Conclusions:Male patients are more prone to phlegm and turbidities than female patients. PCT and D-D were the main risk factors of phlegm-heat obstruction syndrome. Elevated hemoglobin is a risk factor for patients with phlegm stasis and lung syndrome. Heart function classification is the main risk factor of phlegm-dampness-stasis heat syndrome.

Diagnosis and treatment of community-acquired pneumonia in elderly patients combined with coronavirus disease 2019: analysis of 1 case / 中华危重病急救医学

Objective:To explore the pathological characteristics and outcomes of elderly patients with community acquired pneumonia (CAP) accompanied by coronavirus disease 2019 (COVID-19).Methods:The diagnosis and treatment process of one elderly patient with CAP accompanied by COVID-19 who was admitted to COVID-19 Treatment Center of Liaoning Province on February 7, 2020 were reviewed. The experience of treatment by analyzing the characteristics of such type of patients during diagnosis and treatment were summarized.Results:A female patient, aged 79 years ald, was admitted to the Center with following features: fever, dry cough, fatigue with dyspnea, scattered moist rales in both lungs, oxygenation index (PaCO 2/FiO 2) of 95 mmHg (1 mmHg = 0.133 kPa), and diffuse interstitial pneumonia in both lungs indicated by chest CT, of which the majority were ground glass-like and fibrous lesions. It was confirmed to be consistent with the feature of severe COVID-19 cases. The patient was successfully cured one month later following anti-inflammatory, anti-viral and high-flow oxygen therapies, homeostasis maintenance of the body, psychological counseling, etc. Accordingly, the treatment experience in CAP combined with COVID-19 in the elderly patients was summarized as follows. In respiratory system, the timing of high-flow oxygen therapy and mechanical ventilation should be seized. As for anti-inflammatory and antiviral therapy, attention should be paid to the treatment of CAP as well as antiviral therapy and symptomatic and supportive therapy. With the progression of the disease, the production of drug-resistant bacteria and the possibility of fungal infection should be paid attention to. For the circulatory system, we should pay attention to the stability of fluid volume and internal environment, and strengthen hemodynamic monitoring and bedside ultrasound to evaluate the cardiovascular capacity-load. In the aspect of the immune system, the selection of the application time of immune-enhancers and glucocorticoids should be paid attention to. In terms of enteral nutrition, early low-fat and high-protein diet is conducive to the recovery of intestinal function and the prevention of bacterial translocation. In addition to the protection of the function of important organs, therapies such as reasonable sedation and psychological intervention should also be used. Conclusions:Elderly patients with CAP accompanied by COVID-19 have complicated conditions and high degree of difficulty in treatment. Comprehensive evaluation of the disease as well as synthetic and effective intervention are the key factors of successful treatment of such patients.

The effect of Ulinastatin on myocardial cell apoptosis and Caspase-3 channel / 中华急诊医学杂志

Objective To investigate the effect of ulinastatin on myocardial injury in rats with sepsis.Methods Thirty male SD rats were randomly (random number) divided into 3 groups:sham operation group (Sham group,n=10),sepsis group (sepsis group,n=10) and ulinastatin group (UTI group,n=10).The sepsis model of the rats was subjected to cecal ligation and puncture (CLP).Rats of UTI group were given 200 000 U/kg ulinastatin at 6 hour after modeling,and dosing was repeated every 12 h.Blood samples were collected from inferior vena cava at 6,12,24,36 h after modeling for determination of cardiac troponin-Ⅰ (cTnI) and the inflammatory factor by ELISA,then the rats were sacrificed and hearts were removed for myocardial tissue stained with hematoxylin eosin (HE) staining,the expression of Bax/Bcl-2 protein level and myocardial cell apoptosis were detected by TUNEL.The level of caspase-3 protein in myocardial tissue was detected by Western-blot.Results The level of cTnI (ng/mL) in serum in UTI group at 6,12,24 and 36 h after modeling were significantly lower compared with sepsis group(P＜0.05).The protein expression levels of TNF-α and IL-6 (ng/mL) in group UTI were higher than those in Sham group,but was significantly lower than those in Sepsis group (P＜0.05).HE staining showed that inflammatory cell infiltration present in myocardial cells,edema and vacuole formation were observed in sepsis group,while those were significantly attenuated in UTI group compared with sepsis group.UTI increased the level of myocardial Bcl-2 protein in the rats (P＜0.05),and reduced the level of myocardial Bax protein (P＜0.05).TUNEL and HE staining showed apoptosis cells in UTI group was significantly reduced compared with sepsis group [(32.2±4.8)％ vs.(58.4±5.6)％,P＜0.05];Western-blot method showed the level of Caspase-3 protein in UTI group was higher than that in group sham (0.32±0.048) vs.(0.12±0.03),P＜0.05],but significantly lower than that of Sepsis group [(0.32±0.048) vs.(0.55±0.052),P＜0.05].Conclusions Ulinastatin can reduce proinflammatory mediators release in the blood of sepsis rats,and inhibit the apoptosis of myocardial cells protecting myocardial cells through the regulation of the Caspase-3 pathway during sepsis.

Inspiration on building medical emergency clinical teaching platform brought by college student clinic skill competition / 中华医学教育探索杂志

Based on the clinical skill competition of college students of the advanced medical colleges and universities nationwide,we aimed at the analysis on the weaknesses of medical emergency clinical practical teaching and emphasis on the theoretical education and neglect the practical education,and humanistic care,etc.In the clinical practice teaching of emergency,we combined the clinical skill training with physician occupation quality training,pay attention to the practice of advanced simulation exercises,gradual transition,clinical thinking,training medical students hands-on,team cooperation ability and humane accomplishment,to improve their ability of analyzing and solving problems and eventually optimize medical emergency clinical practical teaching,formulate the clinical practice standards as well as promote the reform and innovation of clinical teaching.

Regulation of α-tocopherol on NFκB and Nrf2 signaling pathway at early stage of N-nitrosomethylbenzylamine⁃induced human esophageal cell carcinogenesis / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the regulation of α-Tocopherol on NFκB and Nrf2 signaling pathway at early stage of N-nitrosomethylbenzylamine (NMBzA)-induced human esophageal carcinogenesis.</p><p><b>METHODS</b>Human normal esophageal HET-1A cells were treated with NMBzA at 50 µmol/L, 100 µmol/L for 24 h to intimate the initiation of esophageal carcinogenesis. For intervention groups, HET-1A cells were pre-treated with α-T at 25, 50, 100 µmol/L for 3 h and then co-treated with NMBzA (100 µmol/L) for 24 h. In comparison with HET-1A cells, human esophageal cancer EC109 cells were treated with α-T at corresponding concentrations. Cells treated with 0.1% DMSO were used as negative control. Immunofluorence staining was used for the determination of distribution and activation of NFκB p65 and Nrf2 in the cell. Real time PCR and Western blot were used to determine the expression levels of target genes including cyclinD1, KI67, proliferating cell nuclear antigen (PCNA), cyclo-oxygen-ase 2 (COX2), 5LOX, HO-1, NQO1 and GCLC. Flow cytometry was utilized to analyze the reactive oxygen species contents in the cells.</p><p><b>RESULTS</b>As compared to the control group (1.00 ± 0.08), the expression of CyclinD1 (2.99 ± 0.15), KI67 (2.35 ± 0.38) and PCNA (2.46 ± 0.25) in HET-1A were all markedly increased by NMBzA treatment (F values were 97.23, 65.28, 34.62, P < 0.001). Also, the proportion of cells with nucleus translocation of NFκB p65 (71.0%, 98/138) or Nrf2 (36.3%, 49/135) were significantly increased (χ² values were 194.71, 133.72, P < 0.001), and the expression of COX2 (3.22 ± 0.17), 5LOX (2.87 ± 0.12) as well as HO-1 (1.87 ± 0.22), NQO1 (2.14 ± 0.08), GCLC (2.63 ± 0.41) at protein levels were elevated (F values were 72.35, 43.87, 69.23, 71.34, 85.79, P values were 0.013, 0.015, 0.010, 0.011, 0.002). Under the treatment with 50 µmol/L α-T, comparing with the control group(59.1%,65/110),the nuclear translocation of NFκB p65 (77.7%, 8/104) was clearly inhibited (χ² = 148.1, P < 0.001), and protein expression levels of COX2 (0.74 ± 0.19) and 5LOX (0.42 ± 0.13) were decreased (F values were 56.31, 73.25, P values were 0.003, 0.001). However, no changes on Nrf2 signaling pathway were observed; α-T showed little impact on NFκB or Nrf2 pathway in EC109 cells.</p><p><b>CONCLUSIONS</b>At the early stage of NMBz-induced esophageal cancer, α-T could block the initiation of carcinogenesis through suppressing the activation of NFκB signaling pathway. It might be the major mechanism by which α-T is potentially chemopreventive to esophageal cancer. During the progression of esophageal cancer, the cells may acquire the adaptive functions to accommodate oxidative stress via activating Nrf2 pathway.</p>

Comparison of Three Procedures in the Establishment of Rat Models of Focal Cerebral Ischemia / 中国实验动物学报

@#Objective To compare and evaluate the efficacy of three operating procedures to produce permanent rat models of cerebral ischemia, including mortality, neurological evaluation score, infarction volume of ischemia and time consumed in the operation. Methods The rats were randomly divided into three groups. Group 1: The common carotid artery (CCA) and external carotid artery (ECA) were ligated during the operation and ICA was clipped temporarily by artery clamp. Group 2: The CCA and ECA were ligated and the superior thyroid artery and pterygopalatine artery were exposed but not ligated. Silk suture was utilized to hang the internal carotid artery (ICA). Plastic suture was inserted into ICA and stepped over the initiation point of pterygopalatine artery under microscopic observation. Group 3: The CCA and ECA were ligated and ICA was hanged with a silk suture, and the pterygopalatine artery was not exposed, but during the period of inserting plastic suture, the proximal part of the suture was pressed to make the suture's round distal end elevated, and then pass the initial point of pterygopalatine artery. Mortality, neurological score, volume of ischemic infarction and operation time consumed of the three groups were compared after the operation. Results The procedure to prepare the model was most efficient in the group 3, taking only 17.5 min to complete, significantly less than that in the group 1 (50 min) and group 2 (40 min), (P<0.05), and with a lower mortality and more steady neurological evaluation score and infarction volume. Conclusion The use of the third operating method can shorten the operation time and improve the efficacy of operation. Using this method, more consistent and repetitive focal cerebral ischemia models can be produced effectively, and meet the demands of clinical trials.

Experimental study on the cell culture and secretory function of parathyroid cells in rats / 中国现代普通外科进展

Objective:To investigate the culture technique and secretory function of parathyroid cells in rats in order to establish experimental basis of parathyroid cell transplantation.Methods:After parathyroid glands were digested by collagenase and trypsin,isolated parathyroid cells were cultured.The cells cultured for 5 days were observed by transmission electron microscopy.Parathyroid hormone obtained from the cell culture fluid on day 1,3,5 and 7 was measured respectively.Results:Parathyroid cells cultured for 5 days had abundant rough surfaced endoplasmic reticula、Golgi bodies、stab mitochondria and secretory granules.Cultured parathyroid cells could secrete parathyroid hormone normally,especially on the 5th day.Conclusion:Parathyroid cells cultured for 5 days still possess original morphologic structure and have maximum secreting function,so it is an optimal donor for transplantation.