1.Devolepment of Real-time Fluorescence Quantitative RT-PCR Detection for Avian Influenza Virus H7N9 Type
Journal of Modern Laboratory Medicine 2016;31(3):48-50,54
Objective To establish a rapid and accurate method of fluorescent quantitative PCR detection for Avian Influenza Virus H7N9 type.Methods A pair of primers and TaqMan fluorescent probes were designed specificly for H7N9 gene ac-cording to the published nucleotide sequence from National Center for Biotechnology Information.Positive recombinant plas-mid was built and the minimum copies of detection,repeatability and specificitv were tested.Results The results showed that this assay obtained positive recombinant plasmid,the linear relation was fine in the range from 102 to 108 copies/μl.The minimum copies was 500 copies/μl with this method,specificitv and repeatability(CV<1%)were fine,no cross reaction with other viruses.The precision was 100% (16/16)with this method for detecting 8 positive samples and 8 negative sam-ples.Conclution The establishment methods of fluorescent quantitative PCR detection could be used for the rapid diagnosis Avian Influenza virus H7N9 types.
2.Prick negative electrostatic field(PNESF)and the life spans of mice with ascites tumors
Ti TONG ; Haoyang TONG ; Yu ZHANG ; Yingchun SUN
Chinese Journal of Physical Medicine and Rehabilitation 2008;30(5):299-302
Objective To study the effects of PNESF at different voltages on the life spans of ascites tumor bearing mice.Methods Forty ascites tumor bearing mice were divided into 5 groups and exposed to PNESF at 0,10,17,23 and 30 kV,respectively,for 30 minutes daily on 4 successive days.All of the animals then received routine care until death.Their eating habits,body weights and life spans were observed.A straight line fitting method was used for statistical analysis. Results The body weights of the mice exposed to 17 kV and 23 kV PNESF increased quickly during the four days of PNESF exposure.The weight gain in all groups declined after PNESF exposure was stopped.Thereafter the body weights of the mice exposed to the 17 kV PNESF increased more slowly than those in the control group.The mice exposed to the 23 kV PNESF gained weight more quickly than other groups.The mice exposed to the 17 kV PNESF lived the longest,demonstrating a lengthening of life of 25.5%compared with the control group.Conclusion Exposure to a PNESF of appropriate intensity might significantly increase the life span of ascites tumor bearing mice.
3. Silencing XBP1 expression enhances the sensitivity of human osteosarcoma HOS cells to MPPα-PDT
Tumor 2020;40(1):31-40
Objective: To investigate the role of X-box binding protein 1 (XBP1) in the treatment of human osteosarcoma HOS cells by pyropheophorbide-α methyl ester-mediated photodynamic therapy (MPPα-PDT), and to explore the possible mechanism. Methods: HOS cells were treated by MPPα-PDT for 6, 12 and 24 h, and the expression levels of inositol-requiring enzyme 1 alpha (IRE1α)-XBP1 pathway-related protein IRE1α and XBP1 were detected by Western blotting. The specific siRNA targeting XBP1 gene was transfected into HOS cells by lipofection, and treated by MPPα-PDT. The cells were divided into the blank group, siRNA-negative control (NC) group, siRNA-XBP1 group, MPPα-PDT group and MPPα-PDT+siRNA-XBP1 group. The expressions of XBP1 mRNA and protein were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The proliferation of HOS cells was detected by CCK-8 assay. The apoptotic rate was analyzed by flow cytometry (FCM). The expression levels of cleaved-caspase 3 and cleaved-poly ADP-ribose polymerase (cleaved-PARP) were determined by Western blotting. The intracellular level of reactive oxygen species (ROS) was detected by DCFH-DA probe staining, then the expression levels of Catalase and superoxide dismutase 1 (SOD1) proteins were detected by Western blotting. Results: The expression levels of IRE1α-XBP1 pathway-related proteins IRE1α and XBP1 were increased after the treatment of MPPα-PDT (both P < 0.05 ). siRNA-XBP1 inhibited the expression levels of XBP1 mRNA and protein (both P < 0.01). Silencing siRNA-XBP1 expression inhibited the proliferation activity of HOS cells (P < 0.01), up-regulated the apoptosis rate and the expression level of apoptosis-related protein cleaved-caspase 3 (both P < 0.05), and down-regulated the expression levels of antioxidant enzyme-related proteins Catalase and SOD1 (both P < 0.05). Compared with the MPPα-PDT group, the proliferation activity of HOS cells treated with MPPα-PDT+siRNA-XBP1 was decreased (P < 0.01), the apoptosis rate and the expression levels of cleaved-caspase 3 and cleaved-PARP were increased (all P < 0.05), the intracellular ROS level was up-regulated (P < 0.01), and the expression levels of Catalase and SOD1 were decreased (both P < 0.01). Conclusion: MPPα-PDT can induce the activation of IRE1α-XBP1 pathway in HOS cells. Silencing XBP1 can inhibit the proliferation activity of HOS cells, up-regulate the apoptosis rate, and increase the sensitivity of HOS cells to MPPα-PDT. The mechanism may be related to the up-regulation of intracellular ROS level and the down-regulation of anti-oxidation molecules.
4.Clinical efficacy of endoscopic magnetic compression bilio-enteric anastomosis for the treatment of biliary obstruction after complex abdominal surgery
Yu LI ; Haoyang ZHU ; Hao SUN ; Xuemin LIU ; Xiaogang ZHANG ; Bo WANG ; Yi LYU
Chinese Journal of Digestive Surgery 2020;19(5):544-551
Objective:To investigate the clinical efficacy of endoscopic magnetic compression bilio-enteric anastomosis for the treatment of biliary obstruction after complex abdominal surgery.Methods:The retrospective and descriptive study was conducted. The clinical data of 3 patients with biliary obstruction after complex abdominal surgery who were admitted to the First Affiliated Hospital of Xi′an Jiaotong University between January 2012 and December 2019 were collected. There were 2 males and 1 female, aged from 27 to 57 years, with a median age of 56 years. The 3 patients underwent endoscopic magnetic compression bilio-enteric anastomosis to complete internal drainage of bile ducts after several unsuccessful endoscopic or interventional therapy. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow up using outpatient examination was performed to detect the biliary stent, liver function and patency of anastomotic stoma up to December 2019.Results:(1) Surgical situations: all the 3 patients underwent successful endoscopic magnetic compression bilio-enteric anastomosis, including 2 cases with magnetic compression choledochoduodenal anastomosis and 1 case with magnetic compression choledochojejunostomy. The length of biliary stricture, length and width of magnetic device subsidiary magnet/patent magnet, time of magnetic compression for the 3 patients were 7.1 mm, 3.0 mm, 2.0 mm, 7 mm/8 mm, 6 mm/6 mm, 5 mm/5 mm, 130 minutes, 90 minutes, 75 minutes, respectively. (2) Postoperative situations: the time to biliary tract recanalization after operation for the 3 patients were 15 days, 8 days, 9 days, respectively. None of the patients encountered gastrointestinal perforation, hemorrhage or infection. (3) Follow-up: the biliary stents were inserted into the anastomotic stoma for the 3 patients after biliary tract recanalization, including a catheter of percutaneous transhepatic cholangiodrainage (PTCD) with 12 Fr size, a biliary plastic stent with 7 Fr size, a catheter of PTCD with 14 Fr size, respectively. The biliary stents were removed after 17 months, 2 months, and 6 months from the 3 patients, respectively. The 3 patients were followed up for 40 months, 20 months, and 5 months respectively after removing biliary stents, and the concentration of total bilirubin, concentration of aspartate aminotransferase, concentration of alanine aminotransferase for the 3 patients were 5-19 μmol/L, 25-40 U/L, and 20-35 U/L, respectively. The results of ultrasonic examination or magnetic resonance cholangiopancreatography examination showed that no intra-hepatic bile duct dilation or stricture of choledochojejunostomy was found within the 3 patients. One of the 3 patients was hospitalized for biliary tract infection after 37 months from removing biliary stents, and the results of ultrasonic examination or magnetic resonance cholangiopancreatography examination showed intrahepatic cholelithiasis. The cholelithiasis was removed under endoscopy and stricture of choledochojejunostomy was not found within patient by the endoscopic examination or cholangiography examination.Conclusion:The endoscopic magnetic compression bilio-enteric anastomosis is a safe and feasible technique for the treatment of biliary obstruction after complex abdominal surgery with good long-term effects.
5.Protective role of vasonatrin peptide in hepatic ischemia-reperfusion injury in rats through activation of extracellular signal-regulated kinase signaling pathway
Xin ZHANG ; Yulin ZHU ; Chang LIU ; Haoyang ZHU ; Jun YU ; Yi LYU ; Ge ZHAO
Chinese Journal of Hepatobiliary Surgery 2023;29(2):124-128
Objective:To investigate the protective role of extracellular signal-regulated kinase (ERK) signaling pathway in the process that vasonatrin peptide (VNP) reduces hepatic ischemia-reperfusion injury in rats.Methods:Twenty SD rats, weighting 200-250 g, were randomly divided into four groups and each group has five rats. The four groups were sham operation group (S group), ischemia-reperfusion group (I/R group), VNP group (V group) and PD98059+ VNP group (P+ V group). In the rat model of hepatic warm ischemia and reperfusion, the hepatic artery and portal vein of the left lobe and middle lobe of the liver were clamped with arterial clamp for 45 min followed by reperfusion for 120 min. In the V group, VNP (50 μg/kg) was injected 10 minutes before ischemia. In the P+ V group, PD98059 (2 mg/kg) was injected 20 min before VNP injection followed by VNP administration and I/R treatment. The serum levels of alanine amino transaminase (ALT), aspartate amino transferase (AST), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and the superoxide dismutase (SOD) in liver tissue homogenate and malondialdehyde (MDA) were measured. The histopathology of liver tissue was observed. The contents of p-ERK1/2 were detected by Western blot.Results:Compared with S group, in I/R group and P+ V group the serum levels of ALT [(489.65±11.22), (333.05±24.77) vs. (33.78±4.88) U/L], AST [(651.43±14.99), (503.18±21.48) vs. (154.84±12.32) U/L], TNF-α [(12.83±1.09), (9.64±0.57) vs. (2.11±0.11) ng/L], IL-1β [(7.19±0.62), (5.12±0.22) vs. (1.10±0.49) ng/L], MDA [(8.00±0.88), (5.60±1.01) vs. (2.76±1.29) μmol/mg] increased, while SOD [(54.89±10.60), (68.85±8.33) vs. (126.10±15.63) nmol/mg]decreased (all P<0.05). The histopathology of liver tissue revealed that liver structure damaged more seriously in I/R group and P+ V group. Western blot analysis showed that p-ERK1/2 decreased significantly in I/R group and P+ V group. Compared with I/R group, ALT, AST, MDA, TNF-α and IL-1β decreased significantly and SOD increased significantly in V group (all P<0.05). The histopathology of liver tissue revealed that liver structure was damaged slightly, and p-ERK1/2 increased significantly in V group compared with I/R group ( P<0.05). Conclusion:VNP can significantly reduce hepatic ischemia-reperfusion injury through activation of p-ERK1/2 signaling pathway and inhibition of hepatocyte inflammatory response.
6.Clinical characteristics of limb traumatic osteomyelitis in South China
Xingqi ZHAO ; Haoyang WAN ; Hanjun QIN ; Nan JIANG ; Qingrong LIN ; Yanjun HU ; Jun YANG ; Bin YU
Chinese Journal of Orthopaedic Trauma 2020;22(9):741-745
Objective:To characterize limb traumatic osteomyelitis in a regional trauma center in South China.Methods:The case system at Division of Orthopaedics and Traumatology, Department of Orthopaedics, Nanfang Hospital was searched for the data of confirmed limb traumatic osteomyelitis from January 1, 2010 through September 1, 2019. The clinical items collected were gender, age of onset, cause of injury, injury nature (open or closed), infected site (single site or multiple sites), intraoperative culture of pathogenic microorganisms, number and type of pathogenic microorganisms, and amputation. The above data were sorted out and analyzed statistically.Results:A total of 674 patients suffered limb traumatic osteomyelitis in South China with a male to female ratio of 4.81∶ 1. The limb traumatic osteomyelitis resulted mainly from open injury (64.09%, 432/674). It was mainly caused by a traffic accident (39.51%, 211/534). It was featured by single-site infection (83.68%, 564/674). It affected mainly the lower limbs: the tibia (53.19%, 300/564), the femur (18.97%, 107/564), the calcaneus (11.70%, 66/564) and other foot bones (4.61%, 26/564). The positive rate of intraoperative microorganisms culture was 65.26% (355/544). In the majority of patients (74.65%, 265/355), the traumatic osteomyelitis was caused by infection of a single pathogenic microorganism. The most common single pathogenic microorganism was Staphylococcus aureus (38.11%, 101/265), followed by Pseudomonas aeruginosa (20.00%, 53/265). The rate of amputation related to traumatic osteomyelitis was 3.71% (25/674), and the overall disability rate was 4.45% (30/674).Conclusions:In South China, more males were prone to limb traumatic osteomyelitis. Most cases were secondary to an open fracture and caused by a traffic accident. Traumatic osteomyelitis usually occurred at a lower limb. The most common pathogenic microorganism was Staphylococcus aureus.
7.Metabonomics Combined with Prescription Compatibility Theory to Explore the Lipid-lowering Effect of Qige Decoction Compatibility
Kaixin GUO ; Hui TANG ; Yanfang LI ; Xiaoqing YU ; Haoyang WEI ; Keer HUANG ; Wei CHEN ; Xuehong KE
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(7):971-984
Objective To explore the lipid-lowering effect of Qige Decoction before and after compatibility through the combination of pharmacodynamics and liver metabolomics,and to provide new research strategies for exploring the scientific notation of traditional Chinese medicine compatibility.Methods According to the pharmacodynamic strategy,three groups of drug administration were set up as Qige Decoction group,Astragali Radix-Puerariae Radix group,and Pericarpium Citri Tangerinae group.Four indices of blood lipids,serum biochemical indicators,and liver morphology and pathology were used to evaluate the intervention effect of Qige Decoction on hyperlipidemic rats.Liver metabolomics technology was used to analyze the effects of Qige Decoction on metabolites before and after compatibility,and multivariate statistical analysis was used to evaluate the differences between groups in terms of differential metabolites and metabolic pathways.Results Compared with the model group,the callback abilities of four indices of blood lipid in the Qige Decoction group were higher than those in Astragali Radix-Puerariae Radix group and Pericarpium Citri Tangerinae group,among which the total cholesterol(TC)and triglyceride(TG)levels in the Qige Decoction group decreased(P<0.05).A total of 86 potential biomarkers were identified by liver metabolomics,with 23,13,and 7 metabolites being significantly different in the Qige Decoction group,Astragali Radix-Puerariae Radix group,and Pericarpium Citri Tangerinae group,respectively(P<0.05).Metabolic pathway analysis of 29 specific biomarkers with significant callback effects showed that they were related to glycerophospholipid metabolism,linoleic acid metabolism,α-linolenic acid metabolism,sphingolipid metabolism,arachidonic acid metabolism,and unsaturated fatty acid biosynthesis.Qige Decoction mainly regulates glycerophospholipid and linoleic acid metabolism,and uniquely acts on sphingolipid metabolism.Conclusion Qige Decoction has more lipid-lowering targets after compatibility,with better lipid-lowering effects than the Astragali Radix-Puerariae Radix group and Pericarpium Citri Tangerinae group.This study provides experimental evidence and research strategies for further revealing the scientific notation of traditional Chinese medicine compatibility.
8. Analysis of electrocardiogram and blood related indicators in workers occupationally exposed to mercury in Jiangsu Province from 2016 to 2018
Ping YU ; Haoyang YIN ; Jiadi GUO ; Enmin DING ; Baoli ZHU ; Shuangjing CHEN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2019;37(12):949-952
Objective:
To investigate the health status of occupational mercury workers and reveal the effects of mercury exposure on the cardiovascular system.
Methods:
In June 2019, a total of 2651 mercury workers participated in the occupational health examination between 2016-2018 from a thermometer manufacturing plant and a fluorescent lamp manufacturing plant were included in this study. Then, they were divided into a high-level mercury exposure group (425 workers whose urine mercury concentration >35 μg/g creatinine) and a low-mercury mercury exposure group (2226 workers whose urinary mercury concentration <35 μg/g creatinine) . Mercury concentration in the workplace was also detected. Finally, the results of electrocardiogram (ECG) , blood routine, blood biochemistry and other physical examinations were analyzed. The measurement data of age and exposure years were analyzed by test. Urinary mercury and blood parameters were analyzed by Mann-Whitney nonparametric test. Chi-square test was used for the analyses of gender, ECG abnormality rate and other categorical data.
Results:
The 8-hour weighted average allowable concentration (CTWA) of mercury in the workplace of high-exposure group was 0.002 2-0.152 mg/m3. The abnormal rate of ECG in the high-exposed group (29.6%) was higher than that in the low-exposure group (10.1%) in 2018 (
9.A novel nomogram-based model to predict the postoperative overall survival in patients with gastric and colorectal cancer
Siwen WANG ; Kangjing XU ; Xuejin GAO ; Tingting GAO ; Guangming SUN ; Yaqin XIAO ; Haoyang WANG ; Chenghao ZENG ; Deshuai SONG ; Yupeng ZHANG ; Lingli HUANG ; Bo LIAN ; Jianjiao CHEN ; Dong GUO ; Zhenyi JIA ; Yong WANG ; Fangyou GONG ; Junde ZHOU ; Zhigang XUE ; Zhida CHEN ; Gang LI ; Mengbin LI ; Wei ZHAO ; Yanbing ZHOU ; Huanlong QIN ; Xiaoting WU ; Kunhua WANG ; Qiang CHI ; Jianchun YU ; Yun TANG ; Guoli LI ; Li ZHANG ; Xinying WANG
Chinese Journal of Clinical Nutrition 2024;32(3):138-149
Objective:We aimed to develop a novel visualized model based on nomogram to predict postoperative overall survival.Methods:This was a multicenter, retrospective, observational cohort study, including participants with histologically confirmed gastric and colorectal cancer who underwent radical surgery from 11 medical centers in China from August 1, 2015 to June 30, 2018. Baseline characteristics, histopathological data and nutritional status, as assessed using Nutrition Risk Screening 2002 (NRS 2002) score and the scored Patient-Generated Subjective Global Assessment, were collected. The least absolute shrinkage and selection operator regression and Cox regression were used to identify variables to be included in the predictive model. Internal and external validations were performed.Results:There were 681 and 127 patients in the training and validation cohorts, respectively. A total of 188 deaths were observed over a median follow-up period of 59 (range: 58 to 60) months. Two independent predictors of NRS 2002 and Tumor-Node-Metastasis (TNM) stage were identified and incorporated into the prediction nomogram model together with the factor of age. The model's concordance index for 1-, 3- and 5-year overall survival was 0.696, 0.724, and 0.738 in the training cohort and 0.801, 0.812, and 0.793 in the validation cohort, respectively.Conclusions:In this study, a new nomogram prediction model based on NRS 2002 score was developed and validated for predicting the overall postoperative survival of patients with gastric colorectal cancer. This model has good differentiation, calibration and clinical practicability in predicting the long-term survival rate of patients with gastrointestinal cancer after radical surgery.
10.Development of Motion Unit of Simulated Intelligent Endotracheal Suctioning Robot.
Wenjun TAN ; Cunyi SHEN ; Yu LUO ; Haoyang ZHU ; Tao MA ; Dinghui DONG ; Aihua SHI ; Zhao XUE ; Rongqian WU ; Y LYU
Chinese Journal of Medical Instrumentation 2019;43(1):17-20
A motion unit for sucking robot with a stable motion, convenient operation and process simulation is introduced. The key parameters and process data of the sucking operation were obtained from the clinical work, which provided the basis for the design of the sucking robot motion unit. According to the points of sucking action, robotic thumb, forefinger and metacarpophalangeal joints were used to grip the suction tube, and the servo and arm structure were used to simulate the motion of the wrist and elbow to complete the rotation and push of the sputum suction tube. The feasibility is verified through the advanced sputum suction training model. The movement unit is stable in movement, and can smoothly complete the clamping, feeding, back off protection and rotating tube removal of the sputum suction tube, so as to achieve effective sputum suction.
Catheterization
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Intubation, Intratracheal
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Robotics
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Suction