1.Interaction Between Bruceoside B and Intestinal Flora and Its Inhibitory Effect on Human Lung Cancer A549 Cells
Lingyu SHI ; Wenmin WANG ; Yulin FENG ; Shilin YANG ; Yang WAN ; Daofeng CHEN ; Quan WEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(13):160-166
		                        		
		                        			
		                        			ObjectiveTo explore the interaction between bruceoside B and gut microbiota and the inhibitory activity of its metabolites on human lung cancer A549 cells, and to explore the value of bruceoside B in the treatment of non-small cell lung cancer(NSCLC). MethodBruceoside B was co-incubated with the human gut microbiota under anoxic conditions in vitro, and ultra high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to analyze the metabolic transformation products. Cell counting kit-8(CCK-8) assay was performed to determine the effects of bruceoside B and its metabolites on the proliferation of human lung cancer A549 cells and the half inhibitory concentration(IC50) was calculated. Five healthy male rats were gavaged with bruceoside B(2 mg·kg-1) for 7 days after adaptive feeding. The feces of rats were collected before and after administration. 16S rRNA sequencing was used to assess gut microbiota. ResultBruceoside B was mainly metabolized to brusatol by human gut microbiota, the IC50 of bruceoside B and the conversion product to A549 cells were 1 755.50, 19.57 μmol·L-1, respectively, and the conversion product had a better activity at inhibiting A549 cells proliferation than bruceoside B. Additionally, The results of intestinal flora analysis showed no significant differences in α diversity and β diversity of gut microbiota after administration. In terms of species abundance, at the phylum level, bruceoside B decreased the relative abundance of Actinobacteriota and Proteobacteria, increased the relative abundance of Firmicutes, Patescibacteria and Cyanobacteria. At the genus level, bruceoside B decreased the relative abundance of Staphylococcus, Aerococcus and Psychrobacter, increased the relative abundance of Romboutsia, Lactobacillus, Clostridium sensu stricto 1, Norank-f-norank-o-Clostridia-UCG-014, Turicibacter, Allobaculum and Candidatus Saccharimonas. The results of functional prediction showed that the gut microbiota functional compositions were relatively stable. ConclusionBruceoside B can be deglycosylated by intestinal flora and converted into brusatol, with a significant increase in antitumor activity. The administration of bruceoside B will not cause significant changes in the structure and function of the intestinal flora, resulting in intestinal microecological balance disorders, and the administration appears to be beneficial to the intestinal flora of NSCLC patients. 
		                        		
		                        		
		                        		
		                        	
2.Pien Tze Huang alleviates LPS-stimulated liver Kupffer cell injury by regulating miR-155 expression
Luzhen QIU ; Zhaobin YANG ; Shaozhen HE ; Daofeng HUANG ; Xiaomei CHENG ; Huiping CHEN ; Hao XIA
Chinese Journal of Emergency Medicine 2024;33(4):536-541
		                        		
		                        			
		                        			Objective:To explore the mechanism by which Pien Tze Huang improves liver Kupffer cell damage induced by lipopolysaccharide (LPS) by regulating the expression of miR-155.Methods:LPS induced liver Kupffer cells to establish a cell injury model to simulate septic liver injury. RT-qPCR was used to detect the expression of miR-155 in damaged cells, and RT-qPCR, Western Blot, ELISA and flow cytometry were used to evaluate the inflammatory response and apoptosis of damaged cells. Then we treated LPS-induced Kupffer cells with Pien Tze Huang at different concentrations (0 mg/L, 5 mg/L, 10 mg/L and 15 mg/L), and detected the expression of miR-155 in the cells, the inflammatory response of the cells and Apoptosis rate. MiR-155 was silenced in the cell injury model, and RT-qPCR, Western Blot, ELISA and flow cytometry were used to evaluate the effect of miR-155 on inflammatory response and apoptosis of model cells. Overexpression of miR-155 in damaged cells treated with Pien Tze Huang was used to detect changes in cellular inflammatory response and apoptosis. Data are expressed in the form of mean ± standard deviation, and each group of data is analyzed using t test or one-way analysis of variance.Results:In the LPS-induced liver Kupffer cell injury model, the expression of miR-155 was significantly increased ( P<0.05), the expression levels of pro-inflammatory factors IL-6 and TNF-α were significantly increased, and the anti-inflammatory factor IL-10 was significantly increased. was inhibited ( P<0.05), and the cell apoptosis rate was significantly increased ( P<0.05). After Pien Tze Huang treatment, the expression of miR-155 in damaged liver cells was inhibited ( P<0.05), the levels of cellular inflammatory factors IL-6 and TNF-α were inhibited, and the expression of anti-inflammatory factor IL-10 was promoted ( P<0.05). Inhibit cell apoptosis ( P<0.05). Silencing miR-155 reduced the inflammatory response and apoptosis rate of cells ( P<0.05). Overexpression of miR-155 can reverse the effect of Pien Tze Huang on liver cell injury ( P<0.05). Conclusions:In the model of LPS-induced liver Kupffer cell injury, Pien Tze Huang can inhibite the inflammatory response and apoptosis of cells by inhibiting the expression of miR-155.
		                        		
		                        		
		                        		
		                        	
3.Mining prognostic marker of glioma based on weighted gene co-expression network analysis
Chunyu ZHANG ; Liguo YE ; Long WANG ; Yinqiu TAN ; Fanen YUAN ; Ye TAO ; Qianxue CHEN ; Daofeng TIAN
Journal of Chinese Physician 2021;23(4):529-533
		                        		
		                        			
		                        			Objective:To identify effective biomarkers for glioma patients.Methods:The mRNA expression profiles of 464 glioma patients with complete clinical follow-up information were downloaded from the Chinese Glioma Genome Atlas (CGGA). Weighted gene co-expression network analysis (WGCNA) was used to identify gene modules related to World Health Organization (WHO) grading of glioma, and univariate and multivatiate Cox regression analysis were performed to identify gliomas survival-related genes.Results:In weighted gene co-expression analysis, the module Brown was significantly positively correlated with glioma WHO stage ( r=0.55, P<0.05). In univariate analysis, five genes (TAGLN2, IGFBP2, METTL7B, ARAP3, PLAT) that were most significantly associated with clinical prognosis were selected for multivariate survival analysis, and the prognosis model was established to calculate the risk score. The receiver operating characteristic curve (ROC) confirmed that the risk score had high accuracy in predicting the 1-, 3-, 5-year survival rate of glioma patients. The above survival analysis results were verified in the Cancer Genome Atlas (TCGA) database. Conclusions:We use mRNA expression profiles to establish prognostic markers for gliomas to assess the overall survival of patients with glioma.
		                        		
		                        		
		                        		
		                        	
4.Curative effect of neurolysis and tendon transplantation for treatment of severe thermal crush injuries of the upper limb
Chen WANG ; Pengfei LUO ; Fei HE ; Zhenci CUI ; Yingying LIU ; Li LI ; Dasheng CHENG ; Daofeng BEN
Chinese Journal of Trauma 2020;36(4):347-352
		                        		
		                        			
		                        			Objective:To explore the effect of neurolysis and tendon transplantation in functional reconstruction of the upper limb with severe thermal crush injury.Methods:A retrospective case series study was conducted to analyze the clinical data of 12 patients with thermal crush injuries of the upper limb admitted to Changhai Hospital of Naval Medical University from January 2014 to December 2018. There were 9 males and 3 females, aged 22-54 years (mean, 38 years). The percentage of total body surface area (TBSA) burn ranged from 3% to 8% [(4.9±1.4)%], and wound depth was III degree. According to the damage condition of nerve/tendon and whether there was any dysfunction of the affected limb after wound healing, 12 patients received 2 to 4 times of neurolysis and tendon transplant-related surgeries, with an average surgery of 2.7 times. Among them, a total of 18 times of neurolysis were performed, including 7 times of radial neurolysis, 6 times of median nerve neurolysis and 5 times of ulnar neurolysis, and 14 times of tendon transplantation were done, including 6 times of anastomosis of superficial flexor tendon and long thumb extensor tendon, 5 times of tendon repair transplantation and 3 times of anastomosis of lateral wrist extensor tendon and long thumb extensor tendon. The time interval of each operation was 3-6 months [(4.5±1.0) months]. The Changhai pain ruler, disability of arm-shoulder-hand table (DASH) and joint activity assessment table were assessed before the first operation, 3 months and 6 months after the last operation.Results:All the patients were followed up for 6-12 months (mean, 9.2 months). The score of Changhai pain ruler in the affected limb improved from 3 (2, 3)points before surgery to 1 (0.5, 1)points 3 months after surgery and 1 (0, 1)points 6 months after surgery ( P<0.01). The score of DASH improved from (69.9±2.7) points before surgery to (35.1±1.7) points 3 months after surgery and (33.8±2.0) points 6 months after surgery ( P<0.01). The range of motion score was improved from (1.3±0.5) points before surgery to (2.4±0.5) points 3 months after surgery and (2.8±0.4) points 6 months after surgery ( P<0.01). Conclusion:Neurolysis and tendon transplantation in the treatment of severe thermal crush injuries of the upper limb can alleviate pain in the affected limbs, improve upper limb dysfunction, increase mobility of the palm and upper limb joints, and enhance the quality of life of the patients.
		                        		
		                        		
		                        		
		                        	
5. Debridement combined with vacuum sealing drainage in the treatment of severe infection in abdominal wall due to allogeneic umbilical cord embedded in abdominal wall for immunotherapy
Lianjie ZHANG ; Chen WANG ; Pengfei LUO ; Tiansheng CHEN ; Daofeng BEN
Chinese Journal of Burns 2018;34(8):556-558
		                        		
		                        			 Objective:
		                        			To explore the effect of debridement combined with vacuum sealing drainage (VSD) on the treatment of severe infection in abdominal wall due to allogeneic umbilical cord embedded in abdominal wall for immunotherapy.
		                        		
		                        			Methods:
		                        			From January 2015 to December 2016, 12 patients with severe infection in abdominal wall due to allogeneic umbilical cord embedded in abdominal wall for immunotherapy were admitted to our department. They were conducted with systemic anti-infective treatment, local debridement, and VSD. The wounds were continuously washed for 3 to 5 days after the VSD device installed, with negative pressure value from -16.0 to -12.0 kPa. The VSD device was removed 5 to 7 days later. Continue wound dressing by aseptic ribbon gauze was stuffed in the cavity, and the incision was sutured after the granulation tissue grew well in the cavity.
		                        		
		                        			Results:
		                        			In all patients, allogeneic umbilical cords were completely removed and abdominal infection was cured. The wounds healed well, the sensory function of abdominal was normal, and the activity was not restricted. All the patients were followed up for 3 to 6 months with no reinfection or incisional hernia.
		                        		
		                        			Conclusions
		                        			Embeding the whole allogeneic umbilical cord in abdominal wall for immunotherapy can lead to severe infection in abdominal wall. Abdominal infection can be cured by debridement combined with VSD with good clinical results. 
		                        		
		                        		
		                        		
		                        	
6.Clinical features and diagnosis and treatment of acute fatty liver of pregnancy:a report of 12 cases
Kedi CHEN ; Daofeng YANG ; Guang CHEN
Journal of Clinical Hepatology 2018;34(3):558-561
		                        		
		                        			
		                        			Objective To investigate the clinical features and diagnosis and treatment of acute fatty liver of pregnancy(AFLP). Methods A retrospective analysis was performed for the clinical data of 12 patients with AFLP who were diagnosed and treated in Department of Infec-tious Diseases,Tongji Hospital,Tongji Medical College of Huazhong University of Science and Technology,from April 2012 to March 2017, including general data,clinical manifestations,laboratory markers,imaging examinations,treatment,and prognosis. Results All 12 pa-tients developed AFLP in late pregnancy,and major clinical manifestations included gastrointestinal symptoms,liver failure,jaundice,and coagulation disorder. All patients were given multimodality therapy to protect the liver,improve coagulation,and reduce infection;11 pa-tients underwent cesarean section;6 underwent blood filtration;5 underwent plasma exchange. One patient died,resulting in a mortality rate of 8.3%;5 perinatal infants died,resulting in a mortality rate of 35.7%. Conclusion In patients with AFLP,early diagnosis,timely ter-mination of pregnancy,maximum symptomatic/supportive treatment,and control of infection,as well as the artificial liver support system,is the key to improving the prognosis of mothers and infants.
		                        		
		                        		
		                        		
		                        	
7.Beneficial effects of polysaccharides on "two-hit" acute lung injury and endotoxic fever in rats associated with anti-complementary activities.
Yan LU ; Yun JIANG ; Lijun LING ; Yunyi ZHANG ; Hong LI ; Daofeng CHEN
Acta Pharmaceutica Sinica B 2018;8(2):218-227
		                        		
		                        			
		                        			Thunb. is a traditional herb used for clearing heat and eliminating toxins, and has also been used for the treatment of severe acute respiratory syndrome (SARS). the crude polysaccharides (CHCP) exhibited potent anti-complementary activity through both the classical and alternative pathways by acting on components C3 and C4 of the complement system without interfering with the coagulation system. This study was to investigate the preventive effects of CHCP on acute lung injury (ALI) induced by hemorrhagic shock plus lipopolysaccharide (LPS) instillation (two-hit) and LPS-induced fever in rats. CHCP significantly attenuated pulmonary injury in the "two-hit" ALI model by reducing pulmonary edema and protein exudation in bronchoalveolar lavage fluid (BALF). In addition, it reduced the deposit of complement activation products in the lung and improved oxidant-antioxidant imbalance. Moreover, CHCP administration inhibited fever in rats, reduced the number of leukocytes and restored serum complement levels. The inhibition on the inappropriate activation of complement system by CHCP may play an important role in its beneficial effects on inflammatory diseases. The anti-complementary polysaccharides are likely to be among the key substances for the heat-clearing function of .
		                        		
		                        		
		                        		
		                        	
8.Influence of intravenous injection combined with oral of metoprolol tartrate on left ventricular function and adverse cardiovascular events of patients with acute anterior myocardial infarction
Ying CHEN ; Xiaoxia WANG ; Jin WANG ; Yingqi ZHANG ; Caili HAN ; Daofeng YOU
Chinese Journal of Biochemical Pharmaceutics 2017;37(5):249-251
		                        		
		                        			
		                        			Objective To explore the influence of intravenous injection combined with oral of metoprolol tartrate on left ventricular function and adverse cardiovascular events of patients with acute anterior myocardial infarction.Methods84 cases of Patients with acute anterior myocardial infarction treated in the First Hospital of Hebei Medical University were selected as the study objects, and were divided into vein group and combination group according to drugs-taking modes, 42 cases in each groups.The vein group were treated with intravenous injection of metoprolol tartrate, and the combination group were treated with intravenous injection combined with oral of metoprolol tartrate.Clinical effect, left ventricular function, BP and HR levels, and incidence of adverse cardiovascular events were observed in the two groups.ResultsThe total effective rate of the combination group was 95.24% significantly higher than that of 80.95% in the vein group(P<0.05).After treatment, LVEF was significantly higher than that before treatment in the two groups (P<0.05), which of the combination group was significantly higher than that of the vein group (P<0.05).The levels of LVESD and LVEDD were significantly lower than those before treatment in the two groups (P<0.05), which of the combination group were significantly lower than those in the vein group(P<0.05).After treatment, DBP, SBP and HR were significantly lower than those before treatment in the two groups (P<0.05), which of the combination group were significantly lower than those in the vein group(P<0.05).The incidence of adverse cardiovascular disease was 11.90% in the combination group, and it was no significantly different from that of 16.67% in the vein group (P>0.05).ConclusionIntravenous injection combined with oral of metoprolol tartrate can effectively improve left ventricular function of patients with acute anterior myocardial infarction, and reduce incidence of adverse cardiovascular events.
		                        		
		                        		
		                        		
		                        	
9.To analyze the risk factors of tetany in patients with hyperventilation syndrome
Daofeng YOU ; Qiuge QIAO ; Bing XIE ; Xiangming JIANG ; Ying CHEN
Tianjin Medical Journal 2016;44(6):763-765
		                        		
		                        			
		                        			Objective To explore the risk factors causing tetany in patients with hyperventilation syndrome. Meth?ods A total of 103 patients with hyperventilation syndrome treated in our hospital were included in this study. According to whether there was tetany, patients were divided into tetany group and non-tetany group. Values of gender, age, electrolyte, pH and p(CO2) were analysed between two groups. The factors of P<0.1 were engaged in binary Logistic regression. Logistic regression (Forward Wald) was used to analyze the risk factors of tetany in patients with hyperventilation syndrome. Re?sults In 103 patients there were 70 patients with tetany (68%), 33 patients without tetany(32%). The serum K+, serum phos?phorus and p(CO2) values were significantly lower in tetany group than those of non-tetany group (P<0.01), while the pH value was significantly higher in tetany group than those of non-tetany group (P<0.01). There were no significant differences in gen?der, age, serum Na+, serum Cl-, serum calcium (bound calcium and ionized calcium), ionized calcium and serum Mg2+levels be?tween two groups (P>0.05). It was revealed that the younger age, the lower level of the serum K+, serum phosphorus and p(CO2) were the risk factors of tetany through binary Logistic regression analysis. Conclusion The risk factors of tetany in patients with hyperventilation syndrome include younger age, lower level of serum K+and serum phosphorus and reduced p(CO2).
		                        		
		                        		
		                        		
		                        	
10.Development of a C3c-based ELISA method for the determination of anti-complementary potency of Bupleurum polysaccharides.
Mulu WU ; Hong LI ; Yunyi ZHANG ; Daofeng CHEN
Acta Pharmaceutica Sinica B 2015;5(4):316-322
		                        		
		                        			
		                        			Traditionally, determination of inhibitory potency of complement inhibitors is performed by the hemolytic assay. However, this assay is not applicable to the lectin pathway, thus impeding the understanding of complement inhibitors against the overall function of the complement system. The main objective of our study was to develop a specific enzyme-linked immunosorbent assay (ELISA) as an alternative method to assess the anti-complement activity, particularly against the lectin pathway. By using respective coating substrates against different activation pathways, followed by capturing the stable C3c fragments, our ELISA method can be used to screen complement inhibitors against the classical pathway and the lectin pathway. The inhibitory effect of suramin on the classical pathway, as measured by our hemolytic assay is consistent with previous reports. Further assessment of suramin and Bupleurum polysaccharides against the lectin pathway showed a good reproducibility of the method. Comparison of the lectin pathway IC50 between Bupleurum smithii var. parvifolium polysaccharides (1.055 mg/mL) and Bupleurum chinense polysaccharides (0.98 mg/mL) showed that, similar to the classical and alterative pathway, these two Bupleurum polysaccharides had comparable anti-complementary properties against the lectin pathway. The results demonstrate that the described ELISA assay can compensate for the shortcomings of the hemolytic assay in lectin pathway.
		                        		
		                        		
		                        		
		                        	
            
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