3.Site-directed mutagenesis of human IL-29 and antineoplastic activity of the recombinant human IL-29 variant.
Wei CHEN ; Rong ZHU ; Chunlei GE ; Yuan LU ; Liyun LI ; Fei LI ; Minchen WU
Chinese Journal of Biotechnology 2015;31(5):702-710
To explore the anti-tumor proliferation activity of human interleukin-29 (hIL-29) variant and based on bioinformatics analyzed data of hIL-29, a mutant gene hIL-29(mut33,35) was amplified by site-directed mutagenesis and megaprimer PCR. The hIL-29(mut33,35) was inserted into an eukaryotic expression plasmid pPIC9K and successfully expressed in Pichia pastoris GS115. A recombinant variant protein (rhIL-29(mut33,35)) was purified from the ferment supernatant of the engineering GS115. To observe the antineoplastic activity of the variant rhIL-29(mut33,35), a CCK-8 reagent was used to detect the anti-proliferation effect. Results show that it has strong anti-proliferation effect when acted on liver cancer cell BEL7402, colon cancer cell HCT8 and gastric cancer cell SGC7901. The inhibition ratios of the three tumor cells were (30.99 ± 1.58)%, (22.47 ± 1.37)% and (32.05 ± 2.02)%, respectively. In high dose group, the anti-proliferation effect of the rhIL-29(mut33,35) was stronger than that of wild type rhIL-29 (P < 0.01). This indicates the variant rhIL-29(mut33,35) has potential development value for medicine.
Antineoplastic Agents
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pharmacology
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Carcinoma, Hepatocellular
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pathology
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Cell Line, Tumor
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drug effects
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Humans
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Interleukins
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biosynthesis
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pharmacology
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Liver Neoplasms
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pathology
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Mutagenesis, Site-Directed
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Pichia
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Plasmids
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Polymerase Chain Reaction
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Recombinant Proteins
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biosynthesis
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pharmacology
4.Effect of electro-acupuncture at zusanli (ST36) on the expression of ghrelin and HMGB1 in the small intestine of sepsis rats.
Jian-Nong WU ; Wan WU ; Rong-Lin JIANG ; Mei-Fei ZHU ; Shu LEI ; Bin LU
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(9):1113-1117
OBJECTIVETo explore the expression of Ghrelin and high mobility group protein B1 (HMGB1) in the serum and the intestinal tissue of sepsis model rats, and to evaluate the effect of electro-acupuncture (EA) at Zusanli (ST36) on the expression of HMGB1 and Ghrelin.
METHODSForty-eight male Wistar rats were randomly divided into four groups, i.e., the sham-operation (sham), the cecal ligation and puncture group (CLP), the CLP + EA at Zusanli (ST36) group (EA), and the CLP + Ghrelin receptor blocking agent + EA group (GHSRA), 12 in each group. A sepsis rat model was prepared by CLP. The incision of the abdominal wall was immediately sutured along the ventral midline for rats in the Sham group. In the EA group EA at Zusanli (ST36) was performed 20 min after CLP surgery with the constant voltage (2 - 100 Hz, 2 mA) for 30 min. In the GHSRA group, Ghrelin receptor blocking agent, [D-Arg1, D-Phe5, D-Trp79, Leu11]-substance P (700 nmol/kg), was administered through intravenous injection immediately after CLP, and 20 min later, EA at Zusanli (ST36) was performed in the same way as for rats in the EA group. Blood samples were withdrawn 12 h after CLP. The serum levels of Ghrelin and HMGB1 were detected using ELISA. Ghrelin expressions and the number of Ghrelin immunopositive cell in the jejunum were determined by immunohistochemistry. HMGB1 contents of the jejunum tissue were detected by Western blotting.
RESULTSCompared with the Sham group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly increased and levels of Ghrelin and the expression rate of immunopositive cells significantly decreased in the CLP group (P < 0.05). Compared with the CLP group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly decreased, but levels of Ghrelin and the expression rate of immunopositive cells significantly increased in the EA group (P < 0.05). Compared with the EA group, the number of serum immunopositive cells and the expression of HMGB1 in the jejunum tissue significantly increased in the GHSRA group (P < 0.05), but there was no statistical difference in levels of Ghrelin between the two groups (P > 0.05). The serum level of HMGB1 was negatively correlated with Ghrelin in the Sham group, the CLP group, and the EA group (r = -0. 528, P < 0.01).
CONCLUSIONSEA at Zusanli (ST36) could inhibit the expression of HMGB1 in the jejunum of septic rats, and promote the expression of Ghrelin. The expression of HMGB1 was inhibited by Ghrelin receptor blocking agent, which suggested that the anti-inflammation of EA at Zusanli (ST36) might be associated with Ghrelin.
Animals ; Disease Models, Animal ; Electroacupuncture ; Ghrelin ; metabolism ; HMGB1 Protein ; metabolism ; Jejunum ; metabolism ; Male ; Rats ; Rats, Wistar ; Sepsis ; metabolism
5.Effect of deferoxamine on autophagy induction after blast-induced brain injury in rats
Lijun ZHANG ; Rong HU ; Fei LI ; Hui MENG ; Jiangkai LIN ; Gang ZHU ; Hua FENG
Chinese Journal of Trauma 2015;31(8):748-752
Objective To determine the effect of deferoxamine administration on autophagy in a rat model of blast-induced brain injury.Methods Thirty-nine male SD rats were allotted to shamoperated group,injury group and deferoxamine group with 13 rats in each,according to the random number table.Feeney's method was applied to establish the model.Deferoxamine group received deferoxamine of 100 mg/kg intraperitoneally.Sham-operated and injury group were injected with saline intraperitoneally.All treatments were started two hours postinjury at 12 hour interval for up to 28 days.Hemoglobin,rectal temperature,blood glucose and mortality were detected at 1,3,7,14 and 28 days.Morris water maze was conducted.Rats were killed later for detecting the brain defect volume and level of Beclin 1 at the site of injury.Results There were no significant differences among the three groups with respect to hemoglobin,rectal temperature and blood glucose (P > 0.05).Mortality in injury versus deferoxamine groups did not differ significantly (P > 0.05).Volume of defected brain in deferoxamine group was (115.35 ± 13.70) mm3,smaller than (209.99 ± 16.70) mm3 in injury group (P < 0.05).In Morris water maze test,the time spent in the searching the platform and latency to reach the platform were improved in deferoxamine group compared to those in injury group [(3.13 ± 0.35) vs (2.13 ± 0.64);(36.15 ± 26.63) s vs (110 ± 47.34) s respectively] (P < 0.05).Both immunohistochemisty and western blot showed dramatically increased level of Beclin 1 after injury,but treatment with deferoxamine significantly reduced the Beclin 1 expression.Conclusion Level of Beclin 1 is significantly upregulated after blast-induced brain injury in rats,resulting in elevated autophagy postinjury,but the treatment with deferoxamine is neuroprotective possible by lessening autophagy damage.
6.Mechanism ud protection of electro-acupuncture at Zusanli points(足三星穴) on the apoptosis of thymocytes in rats with severe abdominal infection
Shu LEI ; Rong-Lin JIANG ; Jian-Nong WU ; Mei-Fei ZHU ; Yi-Hui ZHI ;
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2006;0(05):-
Objective To study the influence of electro-acupuncture(EA)at Zusanli points(足三里穴) on the apoptosis of thymocytes in rats with abdominal infection and its mechanism.Methods A total of 40 Sprague-Dawley(SD)rats were randomly divided into four groups,including normal control group,model group,non-acupoint group and Zusanli group.The abdominal infection model of rat was made by cecal ligation and puncture(CLP).After abdominal cavity infection for 36 hours,the apoptosis of thymocytes was observed under electron microscope and light microscope,and the apoptosis ratio of thymocytes was determined by Annexin V-PI method with flow cytometry technique.The content of Bcl-2 protein of thymocytes and concentration of corticosterone in plasma were determined.Results Abdominal infection resulted from CLP could significantly increase the apoptosis of thymocytes and lead to the typical histopathological changes of apoptosis of thymocytes under electron microscope and light microscope.Apoptosis ratios of thyrnocytes in model group[(44.7?3.3)%],non-acupoint group[(42.7?3.0)%]and Zusanli group[(32.6?3.3)%] were significantly higher than the ratio in the control group[(21.2?2.3)%,all P0.05).Abdominal infection resulted from CLP also could reduce the content of Bcl-2 protein of thymocytes.The content of Bcl-2 protein of thymocytes in model group(71.2?5.6),non-acupoint group(73.5?5.9)and Zusanli group(82.4?6.8) were significantly lower than normal control group(95.3?6.3,all P
7.Continuous hemodialysis/hemofiltration for treatment of the critical hemophagocytic syndrome in children
Yucai ZHANG ; Qunfang RONG ; Rongxin CHEN ; Yun CUI ; Fei WANG ; Liang XU ; Yiming ZHU
Chinese Journal of Emergency Medicine 2013;22(7):749-754
Objective To study critical hemophagocytic syndrome (HPS) or macrophage activation syndrome (MAS) presented with multiple organ dysfunction syndrome (MODS) in pediatric intensive care unit (PICU),including clinical features and outcomes In order to explore the effect of bedside continuous hemodialysis/hemofiltration (CBP) as adjuvant treatment for severe HPS/MAS.Methods A total of 19 children with HPS/MAS were hospitalized met the diagnostic criteria for HPS from January,2009 to December,2012.Twelve cases were treated with CBP by continuous venin-venin hemodialysis/hemofiltration (CVVHDF) or high-volume hemofiltration (HVHF) following conventional anti-inflammatory therapy.The replacement liquid dose was 50-75 ml/ (kg · h).The organs function were evaluated and laboratory biomarkers including blood 、electrolytes,ferritin changes were measured before and after CBP treatment.Results Ninteen cases of HPS were acute onset and developed to MODS rapidiy after admission to PICU.The main clinical features were the irregular fever or high fever,hepatosplenomegaly and significant liver damage,nervous system dysfunction and disseminated intravascular coagulation (DIC).Eight cases were death and mortality rate was 42.1%,and all death occurred in those aged less than 3 years old.The mortality rate were 25% (3/12) and 71.4% (5/7) in CBP group and non-CBP group respectively.After CBP for 6-24 hours,the fever returned to normal range and blood electrolytes improved.The serum ferritin,serum alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) reduced significantly.Serum creatinine (sCr),blood urea nitrogen (BUN) level improved.Four cases with acute respiratory distress syndrome (ARDS) improved and the ventilator parameters were downregulated.Conclusions Our findings indicate that HPS/MAS complicated with MODS is life threatening with high mortality rate.CBP therapy can lower the fever within a short time,correct electrolyte imbalance,stable circulatory function,improve the lung,liver,and brain function.It is suggested that CBP may be the potential effective therapy in severe HPS/MAS with MODS in children.
8.Study on the protective effect of Guanxinning Tablet on myocardial ischemia reperfusion injury in rats
Fei HU ; Yili RONG ; Keyan ZHU ; Hong LU ; Cheng CHEN ; Minli CHEN ; Yongming PAN
Chinese Journal of Comparative Medicine 2017;27(5):76-82
Objective To observe the effect of Guanxinning Tablet (GXNT) on myocardial infarction and cardiac autonomic nervous function in rats with myocardial ischemia reperfusion injury (MI/RI).Methods Seventy SD rats were divided into 7 groups randomly (n=10);the sham group, the MI/RI group, 75 mg/kg, 150 mg/kg, 300 mg/kg and 600 mg/kg GXNT groups and 300 mg/kg Compound Danshen Tablets (DST) group.All rats were administered orally for 7 days, and then the MI/RI model was made by ligating the left anterior descending branch of coronary artery in rats.The changes of electrocardiogram were recorded and the electrocardiogram of J points and heart rate variability (HRV) parameters were analyzed.At the end of reperfusion, the myocardial infarct size was measured by using Evans blue and tetrazolium chloride (TTC) double staining, and pathological changes of myocardium were observed by HE staining.The changes of serum lactate dehydrogenase (LDH), creatine kinase (CK), malondialdehyde (MDA) and nitric oxide (NO) levels were also detected.Results Compared with MI/RI group, GXNT and DST groups were significantly reduced myocardial infarct size and inhibited the rising of serum LDH and CK activities (P < 0.05, P < 0.01), and also reduced the total or average value of J point during reperfusion (P < 0.05, P < 0.01).Meanwhile, GXNT and DST groups were markedly increased HRV and serum NO level as well as decreased serum MDA content (P < 0.05, P < 0.01), and improved myocardial tissue pathology.Conclusions GXNT can reduce the myocardial infarction in rats with MI/RI, and also improve the cardiac autonomic nervous function.
9.Noninvasive pulmonary arterial pressure monitoring in the children with severe pneumonia and respiratory failures and its significance
Qunfang RONG ; Yucai ZHANG ; Liang XU ; Rongxin CHEN ; Yun CUI ; Yan ZHU ; Fei WANG
Chinese Journal of Applied Clinical Pediatrics 2015;30(4):271-274
Objective To evaluate the value of noninvasive monitoring of pulmonary arterial pressure in the children with severe pneumonia and respiratory failure.Methods A prospective study was adopted to investigate 69 patients who suffered from severe pneumonia and respiratory failure in Pediatric Intensive Care Unit in Shanghai Children's Hospital from June 2013 to December 2013 were involved in this study,except for heart disease.The pulmonary arterial pressure (PAP) and cardiac function were monitored by using bedside color doppler ultrasound cardiogram,such as PAP,cardiac index (CI),left ventricle ejection fraction(LEFT),and heart early diastolic filling velocity maximum/heart late diastolic filling velocity maximum (E/A ratio).They were divided into 2 groups according to PAP,one group as pulmonary arterial pressure normal group,the other group as pulmonary arterial hypertension(PAH) group,and the impact of the PAP on the prognosis and mechanical ventilation was assessed.Milrinone[0.5 μg/(kg · min)] were given the patients who were combined with pulmonary hypertension,and the PAP and cardiac function before using Milrinone and 24 h,48 h and 72 h after giving medicine was observed.Results Among 69 cases,40 cases were male and 29 cases were female,age ranging from 2 months to 12 years old,and the weight range was (14.3 ± 8.9) kg.The pediatric critical illness score(PICS) was 70.5 ± 9.6,and the pediatric risk of score m ortality Ⅲ was 13.5 ± 5.0.Among 69 cases,46 cases had pulmonary arterial hypertension,38 cases of them experienced mechanical ventilation,and 9 cases died.Among 23 cases who had no pulmonary arterial hypertension,only 8 cases experienced mechanical ventilation.There was a significant difference in the mechanical ventilation rate and mortality between two groups(x2 =15.78,P <0.0l ; x2 =5.18,P < 0.05).The mechanical ventilation time was longer in pulmonary arterial hypertension group (t =3.89,P <0.01).PAP was (58.23 ±5.44) mmHg(1 mmHg =0.133 kPa),(49.10 ±4.69) mmHg,(42.53 ±4.54)mmHg and(35.63 ±4.78) mmHg respectively before and after using Milrinone 24 h,48 h and 72 h in 46 cases with pulmonary arterial hypertension,and the pressure decreased significantly after using medicine (F =67.11,P < 0.01).There was no significant difference in CI,LVEF and E/A(all P >0.05).However,9 cases of them did not show any response to Milrinone,and in the end they couldn't live without mechanical ventilation,they died.Conclusions Noninvasive pulmonary arterial pressure monitoring could be beneficial in judging patient's condition and assessing prognosis of children with severe pneumonia and respiratory failure,and milrinone could decrease PAP.
10.Effects of dietary consumption of monounsaturated fatty acid on control of type 2 diabetes mellitus
jing-fen, ZHU ; fei, DAI ; qing-wen, XIE ; yi, FENG ; rong, SHI
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(03):-
Objective To investigate the effects of dietary consumption of monounsaturated fatty acid (MUFA) on the control of type 2 diabetes mellitus. Methods One hundred and eighty-five patients with type 2 diabetes mellitus from a community in Shanghai were randomly divided into MUFA intervention group (MUFA group,n=125) and control group (n=60). The patients in MUFA group consumed tea oil enriched in MUFA for 3 months,while those in control group consumed normal oil. The serum glucose (fasting plasma glucose and 2-hour postprandial glucose),fasting insulin and blood lipid were examined before intervention and three months after intervention,and the parameters were compared within groups and between groups. Results The serum fasting glucose,fasting insulin,total cholesterol and low density lipoprotein-cholesterol 3 months after intervention were significantly lower than those before intervention in MUFA group (P0.05),while the serum fasting glucose,total cholesterol and low density lipoprotein-cholesterol in MUFA group were significantly lower than those in control group 3 months after intervention (P0.05). Conclusion Dietary consumption of MUFA can improve the glucose and lipid metabolism in patients with type 2 diabetes mellitus.