1.Construction of eukaryotic expression plasmid of human promyelocytic leukemia gene of six transcripts and subcellular location analysis of its recombinant proteins
Chinese Journal of Biologicals 2023;36(7):781-785
Objective To construct eukaryotic expression plasmids of human promyelocytic leukaemia(hPML) gene of six transcripts and analyze the subcellular location of the recombinant proteins.Methods Primers were designed according to the hPML gene sequences registered in GenBank databases.Six transcripts of hPML gene fragments(hPML Ⅰ,Ⅱ,Ⅳ,Ⅴ,Ⅵ and Ⅶ) were amplified by RT-PCR,which were linked to the eukaryotic expression vector pCAGGS respectively.The obtained eukaryotic expression plasmids of six transcripts of hPML gene were transfected into 293T cells respectively and detected for their protein expression by Western blot,while transfected into Vero cells and detected for their subcellular location by indirect immunofluorescence assay(IFA).Results The target gene fragments of the six eukaryotic expression plasmids were consistent with the hPML gene sequences registered in GenBank.All the six recombinant proteins showed specific binding with Myc antibody,among which the recombinant protein hPML Ⅰ,Ⅱ,Ⅳ,Ⅴ and Ⅵ were located in the nucleus and cytoplasm,while the recombinant protein hPML Ⅶ was mainly located in the cytoplasm,rarely in the nucleus.Conclusion The eukaryotic expression plasmids of six transcripts of hPML gene all can be expressed correctly in mammalian cells,and the expressed recombinant proteins were located in nucleus and cytoplasm simultaneously or mainly in cytoplasm.This study provides an experimental basis for subsequent study on the antiviral and other biological functions of recombinant protein hPML.
2.Clinical Analysis of Patients with Primary Intestinal Tumous:A Report of 68 Cases
Jiayu LIN ; Yugang WU ; Qi CHEN
Journal of Chinese Physician 2002;0(S1):-
Objective To explore the diagnosis and surgical treatment of primary intestinal tumors, and improve the level of treatment. Methods Retrospective analysis of the clinical was made on the 68 cases of primary small intestinal tumors confirmed by pathological examination in our department in recent 20 years. HZ Results 34.8%(21/68) was benign tumors in 68 cases, and 69.1%(47/68) was malignancies. The common clinical prevsentations were abdominal pain (69.1%,47/68). gastrointestinal he morrhage (41.1%,28/68) and abdomen mass (13.2%,9/68). The preoperative misdiagnosis rate was 70.5%(48/68) .All the 68 cases performed operation, and no death. The 1 ,2 and 5 years survival rates of malignant tumors were 65.8%,42.1% and 29.3% respectively. Conclusions The clinical presertation of primary small intestinal tumor is non-spectific and the misdiagnosis rate is high. Kinds of diagnosis examinations should be done for the cases whose diagnosis are uncertain, and laboratory examinations should be considered if it is need. The main choice of treatment is surgery and the chemotherapy is necessary for malignant tumors also.
3.Effects of ERK signal transduction pathway on delaying the healing of acetic acid-induced gastric ulcer by Celecoxib in rats
Meirong HE ; Jingqiu LIN ; Yugang SONG
Medical Journal of Chinese People's Liberation Army 1983;0(02):-
Objective To investigate the effects of Celecoxib,a selective COX-2 inhibitor,on the healing of gastric ulcer in rats,and reveal the rale of ERK signal transduction pathway in the mechanism of delaying the healing of gastric ulcer.Methods Forty-eight rats were randomly divided into model group(n=40) and sham operation group(n=8).In the rats in model group acetic acid-induced gastric ulcer was reproduced,while in sham operation group,rats underwent the sham procedure.Eight rats in sham-operation group and eight rats in model group were euthanized three days after the procedure.The other thirty-two rats in model group were divided into two subgroups including Celecoxib group and NS group(n=16).Three days after the procedure,rats in Celecoxib group received a gavage of 0.2% Celecoxib solution,and those in NS group received equal amount of 0.9% NaCl solution.Rats in Celecoxib group and NS group were euthanized on sixth and ninth day after ulcer induction(8 rats at each time point in each group).The effects of Celecoxib on the healing of gastric ulcer were observed.Its effects on the activity of Raf-1 and ERK1/2,and the expression level of two transcription factors c-Fos and c-Jun were also determined by Western blot analysis.Results Nine days after ulcer induction,the ulcer area was 11.9?3.1mm2 and 19.7?3.8mm2 in NS group and Celecoxib group,respectively,and they were much smaller than those on third day(P
4.Expression of HDC and H~+,K~+-ATPase in gastric mucosa during healing of experimental gastric ulcer in rats
Meirong HE ; Jinqiu LIN ; Yugang SONG ; Zhuosheng LAI
Journal of Third Military Medical University 2003;0(14):-
Objective To investigate the expression changes of histidine decarboxylase(HDC)and H+,K+-ATPase in gastric mucosa during the healing of experimental gastric ulcer in rats.Methods The ulcers were caused by applying acetic acid to the serosal surface of the anterior face of the rat gastric body.At different time points during ulcer healing,HDC and H+,K+-ATPase mRNA and protein expressions were studied by using reverse transcription-polymerase chain reaction and Western blot respectively.Results An ulcer crater developed on the anterior face of the gastric body on day 1 after the induction of ulcers,and the ulcer area was biggest on day 3.On day 12,most of the gastric ulcers had healed.Compared with the control group,the HDC and H+,K+-ATPase mRNA expression in the gastric mucosa of ulcer rats showed a decrease on day 1,and increased back to initial level on day 9.The protein expression of HDC and H+,K+-ATPase in gastric mucosa of ulcer rats decreased immediately on day 1,more on day 6,and returned to the initial levels on day 12.Conclusion The mRNA and protein expressions of HDC and H+,K+-ATPase decrease in the healing process of gastric ulcers,resulting in accelerated ulcer healing through inhibiting gastric acid secretion.
5.Flow Injection Analysis of Tannic Acid withEnhanced Chemiluminescent Detection
Shifeng LI ; Hua CUI ; Yugang SUN ; Xiangqin LIN
Chinese Journal of Analytical Chemistry 2001;29(3):333-335
A novel flow injection chemiluminescent enhancement method for the determination of tannic acid based on luminol-KIO4-Mn2+ system is developed. The method is simple, rapid and effective to determine tannic acid in the range of 3.0×10-8~5.0×10-6 mol/L with a detection limit of 9×10-9 mol/L. The relative standard deviation is 1.5% for the determination of 1.0×10-6 mol/L tannic acid (n=11). The mehtod has been applied to determine the content of tannic acid in Chinese gall with satisfactory results.
6.Effect of sevoflurane on myocardial injury induced by hemorrhagic shock and resuscitation in pigs
Li WANG ; Keyan CHEN ; Yugang DIAO ; Lin LI ; Tiezheng ZHANG
Chinese Journal of Anesthesiology 2015;35(9):1065-1067
Objective To evaluate the effect of sevoflurane on myocardial injury induced by hemorrhagic shock and resuscitation (HS/R) in pigs.Methods Twenty-four Bama miniature pigs (12 males, 12 females) , weighing 20-25 kg, aged 3-5 months, were randomly divided into 3 groups (n=8 each) using a random number table: sham operation group (group S) , HS/R group and sevoflurane group (group Sev).The left and right femoral arteries and right femoral vein were cannulated for blood pressure monitoring, blood-letting, blood sampling and fluid infusion.HS/R was induced by blood-letting maintaining for 1 h, followed by resuscitation with autologous blood reinfusion and infusion of lactated Ringer's solution 2 times the volume of the blood withdrawn.The pigs in group Sev were exposed to 2% sevoflurane for 30 min before resuscitation.After cannulation, at 30 min after hemorrhagic shock, before resuscitation, and at 30 min, and 1.5, 2.5 and 3.5 h after resuscitation, blood samples were collected from the femoral artery for determination of creatine kinase-MB (CK-MB) activity and cardiac troponin Ⅰ (cTnI) concentration in serum using an automatic biochemical analyzer.Myocardial specimens were obtained at 3.5 h after resuscitation for detection of tumor necrosis factor-alpha (TNF-ot) and interleukin-6 (IL-6) contents (by ELISA) , and phosphor-signal transducer and activator of transcription 1 (p-STAT1) expression (by Western blot), and for examination of the pathological changes (with light microscope).Results Compared with S group , the CK-MB activity and cTnI concentration in serum and contents of TNF-α and IL-6 were significantly increased, and the expression of p-STAT1 was up-regulated in HS/R and Sev groups (P<0.05).Compared with HS/R group, the CK-MB activity and cTnI concentration in serum and contents of TNF-α and IL-6 were significantly decreased, and the expression of p-STAT1 was downregulated (P<0.05) , and the pathological changes of myocardia were alleviated in Sev group.Conclusion Sevoflurane can alleviate HS/R-induced damage to myocardia of pigs, and inhibited STAT1 activity and attenuated inflammatory responses in the myocardium are involved in the mechanism.
7.Effect of environmental hypothermia exposure on hemodynamics and oxygen metabolism during general anesthesia in a pig model of hemorrhagic shock
Wei DU ; Yugang DIAO ; Jin ZHOU ; Huijuan CAO ; Tiezheng ZHANG ; Lin LI
Chinese Journal of Anesthesiology 2017;37(2):247-250
Objective To evaluate the effect of environmental hypothermia exposure on hemodynamics and oxygen metabolism during general anesthesia in a pig model of hemorrhagic shock.Methods Twelve pathogen-free Bama miniature pigs of both sexes,weighing 20-24 kg,were divided into 2 groups (n=6 each) using a random number table:room temperature group (group RT) and environmental hypothermia group (group EH).The animals inhaled 2% isoflurane for maintenance of anesthesia.The pigs were placed at room temperature (20-22℃) and at low temperature (-10 ℃) in group RT and group EH,respectively.Hemorrhagic shock was induced by withdrawing blood from the right femoral artery within 20 min (30 ml/kg).Before withdrawing blood (T1),immediately after the end of withdrawing blood (T2),and at 1,2,3,4 and 5 h of shock (T3-7),heart rate,mean arterial pressure,mean pulmonary artery pressure,cardiac index and systemic vascular resistance index were recorded.Blood samples were collected from the right femoral artery and internal jugular vein for blood gas analysis,and lactic acid concentrations,hemoglobin,arterial oxygen partial pressure,arterial oxygen saturation,and mixed venous oxygen saturation were recorded.Oxygen delivery index,oxygen consumption index and O2 extraction rate were calculated.Results Compared with group RT,heart rate at T3-7,cardiac index and oxygen delivery index at T5-7,oxygen consumption index at T1-7,O2 extraction rate at T2-6,and lactic acid concentrations at T5-7 were significantly decreased,and mean arterial pressure at T4,5,mean pulmonary artery pressure at T4-7,systemic vascular resistance index at T3-7,and mixed venous oxygen saturation at T2-6 were increased in group EH (P<0.05).Conclusion Environmental hypothermia exposure inhibits cardiac compensatory responses,increases the peripheral vascular resistance,and aggravates oxygen dysmetabolism during general anesthesia in a pig model of hemorrhagic shock.
8.Radiotherapy patients with identification study on continuous quality improvement
Yugang YANG ; Hongzhi QI ; Lin XU ; Jie HAO ; Ge SHANG ; Yongxing BAO
Chinese Journal of Radiation Oncology 2016;25(2):172-174
Objective To study the recogniting patients identity for the safety and reliability of radiotherapy. Methods Through PDCA 4 footwork, namely, plan, do, check, action the technicians in the hospital to improve patients' identity verification.Results After 4 months of PDCA cycle,the patient identity verification qualified rate increase gradually,from 88.17% up to 99.07%,the privacy of patients satisfaction rate rose from 52. 69% to 98. 15%. The patients identification accuracy rate of 100%, technicians working efficiency has been greatly improved. Conclusions The measure of patient identification can improve the working process of radiotherapy for safety and efficiency and can get better privacy protection.
9.Effect of Euphorbia kansui on urination and kidney AQP2, IL-1beta and TNF-alpha mRNA expression of mice injected with normal saline.
Huiyu LI ; Fan LEI ; Yugang WANG ; Xinyue XIAO ; Jun HU ; Xianlong CHENG ; Dongming XING ; Lei HUA ; Ruichao LIN ; Lijun DU
China Journal of Chinese Materia Medica 2012;37(5):606-610
OBJECTIVETo observe the effect of Euphorbia kansui (E. KS) alcohol extracts on urination and kidney-related expressions of mice injected with normal saline and to discuss its impact on kidney.
METHODMice intraperitoneally injected with normal saline were observed for urination and changes in kidney-related histiocytic factors of after intragastrical administration of E. KS and compared with normal mice.
RESULTE. KS alcohol extracts can promote urination of mice injected with normal saline and enhance peripheral serum creatinine, with no obvious pathological change showed in tissue sections. It had a certain effect on reducing AQP2 expression and enhancing TNF-alpha expression.
CONCLUSIONEuphorbia kansui in large dose has a remarkable effect on kidney but may be accompanied with pathological reactions to some extent, especially the dose of 1.2 g x kg(-1). The pathological reactions may be related with increased serum creatinine and TNF-alpha expression.
Animals ; Aquaporin 2 ; genetics ; Euphorbia ; Interleukin-1beta ; genetics ; Kidney ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Plant Extracts ; pharmacology ; RNA, Messenger ; analysis ; Tumor Necrosis Factor-alpha ; genetics ; Urination ; drug effects
10.Correlation between chemokine CX3C ligand 1, CX3C chemokine receptor 1 and heart function grade, prognosis in patients with chronic heart failure
Chun YANG ; Lei LYU ; Yugang YIN ; Lin CHEN
Chinese Journal of Postgraduates of Medicine 2024;47(9):780-785
Objective:To analyze the correlation between chemokine CX3C ligand 1 (CX3CL1), CX3C chemokine receptor 1 (CX3CR1) and heart function grade, prognosis in patients with chronic heart failure (CHF).Methods:The clinical data of 200 patients with CHF from June 2021 to June 2023 in General Hospital of Eastern Theater of the Chinese People′s Liberation Army and Wuhan Asia Heart Hospital were retrospectively analyzed, and all patients received standardized treatment for heart failure. The baseline clinical data were recorded; the levels of CX3CL1 and CX3CR1 were detected by enzyme linked immunosorbent assay; the heart function grade was evaluated by New York Heart Association (NYHA) heart function grade method. The patients were followed up until December 2023, the patients were divided into poor prognosis group (all-cause death and readmission due to heart failure) and good prognosis group based on their prognosis. Pearson method was used for correlation analysis. Multivariate Logistic regression analysis was used to analyze the independent risk factors of poor prognosis in patients with CHF.Results:Among the 200 patients, NYHA heart function grade Ⅰ to Ⅱ was in 80 cases, Ⅲ to Ⅳ in 120 cases. The levels of CX3CL1 and CX3CR1 in patients with NYHA heart function grade Ⅲ to Ⅳ were significantly higher than those in patients with NYHA heart function grade Ⅰ to Ⅱ: (3.34 ± 0.45) mg/L vs. (2.45 ± 0.26) mg/L and (8.71 ± 0.92) mg/L vs. (2.53 ± 0.35) mg/L, and there were statistical differences ( t = 15.99 and 57.34, P<0.01). The proportion of age<60 years old, rate of coronary heart disease, CX3CL1, CX3CR1, body mass index and high-sensitivity C-reactive protein in poor prognosis group (40 cases) were significantly higher than those in good prognosis group (160 cases): 82.50% (33/40) vs. 10.62% (17/160), 90.00% (36/40) vs. 68.12% (109/160), (3.26 ± 0.77) mg/L vs. (2.25 ± 0.27) mg/L, (8.35 ± 2.01) mg/L vs. (2.48 ± 0.31) mg/L, (26.80 ± 3.55) kg/m 2 vs. (24.74 ± 2.76) kg/m 2 and (9.31 ± 2.19) mg/L vs. (3.58 ± 2.28) mg/L, the rate of smoking history and left ventricular ejection fraction were significantly lower than those in good prognosis group: 37.50% (15/40) vs. 46.88% (75/160) and (30.14 ± 5.77)% vs. (59.40 ± 6.58)%, and there were statistical differences ( P<0.01). Pearson correlation analysis result showed that the CX3CL1 and CX3CR1 were positively correlated with NYHA heart function grade ( r = 0.29 and 0.34, P<0.05), and negatively correlated with prognosis ( r = - 0.54 and - 0.36, P<0.05). Multivariate Logistic regression analysis result showed that the CX3CL1 and CX3CR1 were the independent risk factors of poor prognosis in patients with CHF ( OR = 2.110 and 1.566, 95% CI 0.445 to 3.125 and 0.270 to 3.455, P<0.01). Conclusions:The CX3CL1 and CX3CR1 are closely related to the heart function grade in patients with CHF. At the time of CHF patient admission, it may be considered to combine the two indicators for preliminary evaluation of and provide targeted interventions to improve prognosis.