Effect of Euphorbia helioscopia Aqueous Extract on Chronic Obstructive Pulmonary Disease and Precancerous Lesion-associated Proteins of Lung Tissues in Mice Induced by Cigarette Smoke / 中国实验方剂学杂志

ObjectiveTo investigate the protective effect and mechanism of Euphorbia helioscopia aqueous extract （EHE） on mice with chronic obstructive pulmonary disease （COPD） and its influence on precancerous lesion-associated proteins in lung tissues induced by cigarette smoke （CS）. MethodThe COPD model was induced by CS in 60 mice and the model mice were randomly divided into control group， model group， positive drug group （dexamethasone， 2 mg·kg-1）， and low-， medium-， and high-dose EHE groups （1.875， 3.75， 7.5 g·kg-1）. The high-performance liquid chromatography （HPLC） method was used to determine the related components in EHE. The changes in end-expiratory pause （EEP）， airway resistance （Penh）， expiratory flow at 50% vital capacity （EF50）， and other pulmonary function indexes were detected by the spirometer. The levels of inflammatory factors， such as interleukin （IL）-2， IL-5， IL-18， IL-17A， and IL-27 in bronchoalveolar lavage fluid （BALF） of mice were detected by high-throughput liquid protein chip technology. Hematoxylin-eosin （HE） staining was used to detect the pathological changes in lung tissues in mice. The content of malondialdehyde （MDA）， myeloperoxidase （MPO）， and glutathione peroxidase （GSH-Px） in lung tissues was determined by the colorimetric method. The mRNA relative expression of tumor necrosis factor-α （TNF-α）， transforming growth factor-β （TGF-β）， matrix metalloproteinase-2 （MMP-2）， matrix metalloproteinase-9 （MMP-9）， and matrix metalloproteinase-12 （MMP-12） was detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Immunohistochemistry （IHC） was used to detect the expression of tumor protein （P53） and cell proliferation-associated antigen （Ki67） in lung tissues， and Western blot was used to detect the relative expression of tumor suppressor protein （P16）， DNA （cytosine-5）-methyltransferase 1 （DNMT1）， and fragile histidine triad （FHIT） in lung tissues. ResultThe results showed that the main compounds in EHE included phenols （gallic acid and protocatechuic acid） and flavonoids （such as hyperoside， rutin， myricetin， naringenin， quercetin， luteolin， kaempferol， and licorice chalcone A）， among which gallic acid and rutin were the highest in content. Compared with normal group， model group showed increased levels of EEP， EF50， and Penh （P<0.05）， and showed increased MDA and MPO levels （P<0.01） and decreased GSH-Px （P<0.01）， and the model group displayed increased levels of IL-2， IL-5， IL-18， IL-17A， IL-27， TNF-α， TGF-β， MMP-2， MMP-9， and MMP-12 （P<0.05）. And the model group exhibited up-regulated expression of P53， Ki67， and FHIT in lung tissues （P<0.01） and down-regulated expression of DNMT1 and P16 （P<0.01）. Compared with model group， the EHE groups showed decreased EEP and EF50 levels （P<0.05）. The pathological injury of lung tissues in mice of the model group was observed under HE staining， and the pathological injury of basal cell hyperplasia of lung tissues was gradually improved after treatment with EHE. The EHE groups showed reduced levels of MDA and MPO （P<0.01） and increased GSH-Px （P<0.01）. The EHE groups displayed decreased levels of IL-2， IL-5， IL-18， IL-17A， IL-27， TNF-α， TGF-β， MMP-2， MMP-9， and MMP-12 （P<0.05）. And the EHE groups showed down-regulated Ki67 and FHIT in lung tissues （P<0.05） and up-regulated expression of P53 and DNMT1 （P<0.05）. ConclusionEHE can protect mice from COPD and inhibit precancerous lesions， and the mechanism may be related to the inhibition of inflammation and oxidative stress response， regulation of protease and antiprotease imbalance， and regulation of epithelial cell growth.

Pilot study of the relationship between clinical classification of gallbladder cancer and prognosis: a retrospective multicenter clinical study / 中华外科杂志

Objectives@#To propose a novel clinical classification system of gallbladder cancer, and to investigate the differences of clinicopathological characteristics and prognosis based on patients who underwent radical resection with different types of gallbladder cancer.@*Methods@#The clinical data of 1 059 patients with gallbladder cancer underwent radical resection in 12 institutions in China from January 2013 to December 2017 were retrospectively collected and analyzed.There were 389 males and 670 females, aged (62.0±10.5)years(range:22-88 years).According to the location of tumor and the mode of invasion,the tumors were divided into peritoneal type, hepatic type, hepatic hilum type and mixed type, the surgical procedures were divided into regional radical resection and extended radical resection.The correlation between different types and T stage, N stage, vascular invasion, neural invasion, median survival time and surgical procedures were analyzed.Rates were compared by χ2 test, survival analysis was carried by Kaplan-Meier and Log-rank test.@*Results@#Regional radical resection was performed in 940 cases,including 81 cases in T1 stage,859 cases in T2-T4 stage,119 cases underwent extended radical resection;R0 resection was achieved in 990 cases(93.5%).The overall median survival time was 28 months.There were 81 patients in Tis-T1 stage and 978 patients in T2-T4 stage.The classification of gallbladder cancer in patients with T2-T4 stage: 345 cases(35.3%)of peritoneal type, 331 cases(33.8%) of hepatic type, 122 cases(12.5%) of hepatic hilum type and 180 cases(18.4%) of mixed type.T stage(χ2=288.60,P<0.01),N stage(χ2=68.10, P<0.01), vascular invasion(χ2=128.70, P<0.01)and neural invasion(χ2=54.30, P<0.01)were significantly correlated with the classification.The median survival time of peritoneal type,hepatic type,hepatic hilum type and mixed type was 48 months,21 months,16 months and 11 months,respectively(χ2=80.60,P<0.01).There was no significant difference in median survival time between regional radical resection and extended radical resection in the peritoneal type,hepatic type,hepatic hilum type and mixed type(all P>0.05).@*Conclusion@#With application of new clinical classification, different types of gallbladder cancer are proved to be correlated with TNM stage, malignant biological behavior and prognosis, which will facilitate us in preoperative evaluation,surgical planning and prognosis evaluation.

Comprehensive Proteomic Profiling of Patients' Tears Identifies Potential Biomarkers for the Traumatic Vegetative State / 神经科学通报·英文版

The vegetative state is a complex condition with unclear mechanisms and limited diagnostic, prognostic, and therapeutic methods. In this study, we aimed to explore the proteomic profile of tears from patients in a traumatic vegetative state and identify potential diagnostic markers using tears-a body fluid that can be collected non-invasively. Using iTRAQ quantitative proteomic technology, in the discovery phase, tear samples collected from 16 patients in a traumatic vegetative state and 16 normal individuals were analyzed. Among 1080 identified tear proteins, 57 were upregulated and 15 were downregulated in the patients compared to the controls. Bioinformatics analysis revealed that the differentially-expressed proteins were mainly involved in the wound response and immune response signaling pathways. Furthermore, we verified the levels of 7 differentially-expressed proteins in tears from 50 traumatic vegetative state patients and 50 normal controls (including the samples used in the discovery phase) using ELISA. The results showed that this 7-protein panel had a high discrimination ability for traumatic vegetative state (area under the curve = 0.999). In summary, the altered tear proteomic profile identified in this study provides a basis for potential tear protein markers for diagnosis and prognosis of the traumatic vegetative state and also provides novel insights into the mechanisms of traumatic vegetative state.

Effects of norcantharidin on lipopolysaccharide-induced hepatocyte injury in vitro / 中南大学学报(医学版)

OBJECTIVE@#To study the effects of norcantharidin (NCTD) on lipopolysaccharide (LPS)-induced hepatocyte injury and the expression of TNF-α and IL-6 in vitro.@*METHODS@#Hepatocytes were isolated from male Sprague-Dawley rats by collagenase perfusion. LPS at concentration of 40 mg/L was used to induce injury to the cultured cells, and NCTD (0.5, 1.0, 2.5 μg/mL) was added at the same time. After 24 h of incubation, the cell proliferation rates were detected by MTT. LDH, TNF-α and IL-6 were measured by appropriate reagent kits.NF-κB DNA binding activity was measured.@*RESULTS@#40 mg/L LPS caused a 27% growth inhibition in primary hepatocytes. LDH leakage was 20- fold higher in NCTD-treated hepatocytes than in normal ones. TNF-α and IL-6 expression significantly increased. In cells treated with NCTD at doses of 0.5, 1.0 and 2.5 μg/mL, LDH leakage, TNF-α and IL-6 expression, and NF-κB DNA binding activity were attenuated in a dose dependent manner.@*CONCLUSION@#NCTD protects hepatocytes from injury induced by LPS; the protection is associated with suppression of the inflammatory cytokine TNF-α and IL-6.