1.Application of metabolomics in research on biomarkers for hepatocellular carcinoma
Jiangang WANG ; Huacheng LI ; Xinying FEI
Journal of Clinical Hepatology 2014;30(8):838-840
As a novel research tool,metabolomics technology can reveal the differences in metabolic profiles during the development and progression of hepatocellular carcinoma (HCC),so it has been widely applied for research on HCC biomarkers.The significance of metabo-lomics in the diagnosis of HCC is briefly described,and the metabolomics research aiming at the discovery of HCC biomarkers,including an-imal experiments and clinical studies of metabolites in the serum,urine,and liver tissue,is reviewed.It is pointed out that analyzing and monitoring metabolites in the development and progression of HCC is of great significance for individualized treatment.
2.Effects of biliverdin on lung injury induced by brain death in rats
Juan WANG ; Huacheng ZHOU ; Yulong BO ; Nan ZHANG ; Wenzhi LI
Chinese Journal of Anesthesiology 2010;30(5):615-618
Objective To investigate the effects of exogenous biliverdin on lung injury induced by brain death (BD) in rats. Methods Twenty-three adult male Wistar rats in which Fogarty balloon catheter was successfully inserted into cranial cavity were randomly divided into 3 groups: group Ⅰ sham operation (group S,n = 7); group Ⅱ brain death (group BD, n = 8) and group Ⅲ biliverdin + BD (group B, n = 8). The animals were anesthetized, intubated and mechanically ventilated. Femoral artery and vein were cannulated for MAP monitoring and drug and fluid administration. Brain death was induced by injecting slowly normal saline into the balloon in group Ⅱ and Ⅲ. BD was confirmed by dilated and fixed pupils, apnea, transient hypertension and EEG changes. In group Ⅲ biliverdin 35 mg/kg was injected intraperitoneally as soon as BD was confirmed. The animals were mechanically ventilated for another 1.5 h during which MAP was maintained at 80-120 mm Hg by iv norepinephrine infusion. Arterial blood samples were obtained before anesthesia, immediately before and at 5, 30,60, 90 min after intraperitoneal biliverdin for blood gas analysis and determination of plasma bilirubin concentration. PaO2/FiO2 was calculated. The animals were sacrificed at 1.5 h after biliverdin administration. The left lung was removed for detection of MDA content, SOD activity, total antioxidant capacity, cell apoptosis and biliverdin reductase expression in lung tissue. Results Brain death significantly decreased PaO2/FiO2, lung SOD activity and total antioxidant capacity and increased lung MDA content and apoptosis as compared with sham operation group. IP biliverdin significantly attenuated BD-induced lung injury in group B as compared with group BD. The plasma bilirubin concentration and biliverdin reductase expression were significantly higher in group B than group BD. Conclusion Exogenous biliverdin can attenuate BD-induced lung injury by inhibiting pulmonary oxidative stress response and apoptosis.
3.Effects of inhalation of different concentrations of carbon monoxide on brain death-induced lung injury in rats
Huacheng ZHOU ; Jinfeng LIU ; Wenzhi LI ; Di JIN ; Peng PAN ; Wengang DING
Chinese Journal of Anesthesiology 2010;30(8):976-979
Objective To investigate the effects of inhalation of different concentrations of carbon monoxide (CO) on brain death (BD)-induced lung injury in rats. Methods Thirty-two pathogen free adult male Wistar rats weighing 250-300 g were randomly divided into 4 groups ( n= 8 each): group Ⅰ sham operation (group S);group Ⅱ brain death (group BD) and group Ⅲ and Ⅳ BD + CO 0.025% and 0.050% (group C1, C2 ). The animals were anesthetized and tracheally intubated. Fogarty catheter was inserted into the skull. BD was induced by inflating the balloon slowly at 20 μl/min until apnea developed. The animals were then mechanically ventilated (VT 10 ml/kg, RR 50 bpm, PEEP 2 cm H2O) with 40% O2 in N2 . In group Ⅲ and Ⅳ CO 0.025% and 0.050%were added to the air mixture respectively. In group S the balloon was not inflated. BD was confirmed by apnea,dilated pupils and flat EEG. In group BD,C1 and C2, MAP was maintained at 80-120 mm Hg by norepinephrine infusion. The arterial blood gas analysis was performed before (baseline) and immediately after BD was confirmed (T1) and at 30, 60, 90 and 120 min (T2-5) of CO inhalation. The animals were then sacrificed. The plasma concentrations of IL-6 and TNF-α and the activity of myeloperoxidase (MPO) in the lungs were measured. The W/D lung weight ratio and lung injury score (LIS) were recorded. Results BD significantly decreased PaO2/FiO2, BE and pH while increased plasma IL-6 and TNF-α concentrations, MPO activity in the lungs, the W/D ratio and lung injury score as compared with group S. CO inhalation ameliorated the deleterious effects induced by BD. The antiinfiammatory effect of 0.050% CO was better than that of 0.025 % CO. Conclusion Inhalation of 0.025 % or 0.050% CO can ameliorate BD-induced lung injury in rats, but there is no significant difference in the efficacy.
4.Construction and identification of differential expression genes of peripheral blood cells in radon-exposed mice
Rui CHEN ; Jianxiang LI ; Jihua NIE ; Minhua SHI ; Huacheng HU ; Jian TONG
Chinese Journal of Radiological Medicine and Protection 2009;29(1):17-19
Objective To screen and identify the differential expression genes on peripheral blood cells of mice based on the experimental animal model of radon exposure. Methods BALB/c mice were exposed in a type HD-3 multifunctional radon-room, with the accumulative doses of radon-exposure group at 105 WLM and control group at 1 WLM. Total RNA was extracted from peripheral blood cells and the methods of SMART for dscDNA synthesis and SSH for gene screening was applied. With the construction of the cDNA library enriched with differentially expressed genes, the pMD 18-T plasmid containing LacZ operator at the multiple cloning site was used to allow a blue-white screening. The TA clones were amplified by nested PCR and the reverse Northern blot was used to identify up and down regulation of the clones. The differently expressed cDNA was then sequenced and analyzed. Results The subtracted cDNA libraries were successfully constructed. A total of 390 recombinant white colonies were randomly selected. Among the 312 cDNA monoelones selected from bath forward- and reverse-subtracted libraries,41 clones were chosen to sequence for their differential expressions based on reverse Northern blot. Among the 41 sequenced clones, 10 clones with known function/annotation and 3 new ESTs with the GenBank accession numbers were obtained. Most of the known function/annotation genes were revealed to be related with cell proliferation, metabolism, cellular apoptosis and carcinogenesis. Conclusions The animal model of radon exposure was established and the cDNA library of peripheral blood cells was suceessfully constructed. Radon exposure could up- and down-regulate a series of genes. Differentially expressed genes could be identified by using SSH technique and the results may help exploring mechanisms of random exposure.
5.Clinical analysis of 4 cases of Leigh syndrome in children
Lihui WANG ; Huacheng ZHENG ; Huafang YANG ; Ling YUE ; Yuexian ZUO ; Baoguang LI ; Xiaopu CUI
Journal of Clinical Pediatrics 2016;(2):111-114
Objective To explore the clinical manifestation, diagnosis and prognosis of Leigh syndrome in children. Method Clinical data from 4 cases of Leigh syndrome conifrmed by genetic testing were retrospectively analyzed. The related literature were reviewed. Results In 4 cases, 3 were boys and one was a girl, 3 cases were onset in infant and one case was in school age. The main manifestations were mental retardation, low muscle tone, convulsions, feeding dififculties, drooping eyelids, extraocular muscle paralysis and nystagmus, irritation, activity intolerance etc. The brain magnetic resonance imaging (MRI) revealed symmetry long T1, T2 abnormal signal in brainstem, bilateral globus pallidus, thalamus, cerebellar dentate nuclei, and periaqueductal, 3 cases involved midbrain, one case involved thalamus, and one case involved cerebellar dentate nuclei;2 cases had encephalatrophy. Electromyography was normal in all cases. The levels of lactate in blood and cerebrospinal lfuid were increased. Mitochondrial DNA (mtDNA) detection found the mutation of mtDNA 8993 T>G in one case, and the mutation of mtDNA 9176 T>C in another 3 cases. The case onset in school age died of respiratory failure one month later, and another 3 cases were still in follow up, there were mental retardation, but no signiifcant setback. Conclusion The clinical manifestations of Leigh syndrome in children are diverse. The diagnosis is based on the typical clinical manifestations and MRI, blood and/or cerebrospinal lfuid lactate levels. The genetic testing is the golden standard for diagnosis.
6.Correlation between CYP2J2 gene polymorphism and coronary heart disease in Chinese Han population and effects of CYP2J2 geneover-expressionon aortic smooth muscle cells of ApoE-/-mice
Rui LI ; Jian QIU ; Lei SHI ; Yuan ZHANG ; Huacheng YAN ; Yunjun RUAN ; Hua XIAO
The Journal of Practical Medicine 2015;31(23):3831-3836
Objective To investigate the relationship between CYP2J2*7 mutation(G-76T) and coronary heart disease (CHD) in Chinese Hanpopulation and to study the effects of CYP2J2 geneover-expressionon the proliferation and migrationof aortic smooth muscle cells of ApoE-/- mice. Methods CYP2J2*7 genotype was detectedin 500 patients with CHD and 478 controlsubjects by the Polymerase Chain Reaction-Restriction Frag-ment Length Polymorphism (PCR-RFLP). Culturedaortic smooth muscle cells of ApoE-/- mice were divided into control group, sham transfectiongroup and CYP2J2 over-expression group. Cell proliferation and migration were investigated after CYP2J2 over-expressionby MTS and Transwell assay. Results The frequency of CYP2J2*7 in CHD group was significantly higher than that incontrol group (10.00% vs. 6.49%, P = 0.046). Same is the case in female cases(P = 0.026). Compared with these of aortic smooth muscle cells incontrol group and sham trans-fectiongroup, the cell proliferation in 24, 48, 72 h, and the cell migration in 48 h after CYP2J2 over-expression in CYP2J2 group were significantly suppressed. Conclusions CYP2J2*7 mutation might increase the risk of CHD in Chinese Han population. CYP2J2 over-expression can suppress the proliferation and migration of aortic smooth muscle cells and CYP2J2 might have the effect of anti-atherosclerosis.
7.Influence of Gleevec on expression of miR-146a,miR-29b and DNMT in K562 cells
Lina WANG ; Jianming ZENG ; Huacheng WANG ; Mo LI ; Yifei LONG ; Guangyuan DENG ; Cha CHEN
Chongqing Medicine 2014;(3):301-303
Objective To investigate the changes of microRNA (miR)-146a ,miR-29b expression levels and the 3 kinds of meth-ylase DNMT1 ,DNMT3a and DNMT3b levels in K562 cell lines after BCR/ABL inhibitor Gleevec treatment .Methods The half maximal inhibitory concentration(IC50 ) of Gleevec on K562 cells was detected by the MTT method .The stem loop primers method and the fluorogenic quantitative PCR were adopted to detect miRNAs and the methylase gene level .Results IC50 of Gleevec acting on K562 cells was 40 .85μmol/L .After Gleevec action ,miR-29b showed the increasing trend ,but 3 kinds of methylase expression level were decreased to some extent .Gleevec could significantly increase the miR-146a level in K562 cells(P<0 .05) .Conclusion Gleevec can influence the expression of miR-146a ,miR-29b and DNMTs levels in K562 cells .
8.Relationship between concentration of exhaled pentane and degree of lung injury in non-heart-beating rabbits
Yuting WEI ; Huacheng ZHOU ; Jingchun XING ; Zhijie FU ; Jinfeng LIU ; Wenzhi LI
Chinese Journal of Anesthesiology 2012;32(7):857-859
Objective To evaluate the relationship between the concentration of pentane in the exhaled air and degree of the lung injury in non-heart-beating (NHB) rabbits.Methods Twenty-four healthy male Japanese white rabbits weighing 2.4-3.0 kg were randomly divided into 4 groups ( n =6 each):A,B,C and D groups.The NHB model was established by exsanguination through the femoral artery.The exhaled gases were collected and lung tissues were removed at 0,30,60 and 120 min after cardiac arrest in A,B,C and D groups respectively.The concentration of pentane in the exhaled gases was detected immediately using the gas chromatography-mass spectrography.The wet to dry (W/D) lung weight ratio and content of malondialdehyde (MDA) in lung tissues were measured.The lung injury score (LIS) was recorded.The maximal volume ( Vmax ) of the lung was recorded when the airway pressure reached 30 cm H2O.Results Compared with groups A and B,the exhaled pentane concentration was significantly increased in group C,and the W/D ratio,content of MDA and LIS were significantly increased,while Vmax was significantly decreased in group D ( P < 0.05).Compared with group C,W/D ratio and LIS were significantly increased in group D ( P < 0.05 ).Conclusion The concentration of exhaled pentane can not reflect the degree of the lung injury in NHB rabbits.
9.Effects of Ternatolide on expression granulysin mRNAin the peripheral blood lymphocytes patients with mycobacterium tuberculosis
Li ZHAN ; Huacheng DAI ; Zhiping YANG ; Zhuwen YI ; Ping SHU ; Shiming CHENG ;
Chinese Pharmacological Bulletin 1986;0(04):-
AIM To study the effects of ternatolide(tern) on expression of granulysin mRNA in the activated peripheral blood lymphocytes(PBLS) of the adults and children with pulmonary Mycobacterium tuberculosis(PMTB). METHODS Using competitive quantitative reverse transcription polymerase chain reaction(QC TR PCR)analyze granulysin gene expression in the PBLs. RESULTS The dosage of 100 mg?L -1 , 200mg?L -1 of ternatolide significantly enhanced the granulysin mRNA experession in PBLs in vitro stimulated phytohemagglutinin (PHA) from adults and children with PMTB (P0 05). CONCLUSION The molecular mechanism of tern. against intracellular pathogens might be associated with the inducting highly level on the expression of granulysin mRNA, a anti bacterial peptide in activated human cytotoxic lymphocytes.
10.Clinical effect of arthroscopic reduction and suture for tibial intercondylar eminence fractures by perforated Kirschner wire
Fei LIU ; Huacheng HOU ; Li JU ; Fengyong MAO ; Kai TANG
Chinese Journal of Applied Clinical Pediatrics 2023;38(9):693-697
Objective:To investigate the clinical effect of arthroscopic reduction of tibial intercondylar ridge fractures in children using a 2.0 mm perforated Kirschner wire to establish bone tunnels and the Krackow-" 8" suture.Methods:From January 2018 to December 2020, 24 children (14 males and 10 females, aged 6-15 years) with tibial intercondylar ridge fractures admitted to Children′s Hospital of Nanjing Medical University were retrospectively recruited for analyses.All patients were treated with arthroscopic reduction, non-absorbable Krackow-" 8" suture for the base of the anterior cruciate ligament, and establishment of bone tunnels by 2.0 mm perforated Kirschner wire from the proximal tibial epiphysis for inserting a suture to fix the fractures.The affected limb was immobilized with a cast for 4 weeks, followed by active exercises postoperatively.Knee function was assessed using the Lysholm score and IKDC 2000 subjective score.Knee stability was evaluated using the Lachman test.X-rays were taken at 3 and 6 months postoperatively to evaluate fracture healing.Growth retardation of epiphyseal plate at 2 years of follow-up was assessed by bilateral knee X-rays.Results:All the 24 cases were successfully operated, with the operation time of (63.1±20.2) (40-115) min.All children were followed up for 24-36 months[(28.7±3.4) months]. All children achieved an anatomical reduction of the fracture postoperatively.No complications like fracture displacement, suture rupture, or infection were reported.All fractures healed completely at 3 months postoperatively.At the last follow-up, the Lysholm score and IKDC 2000 subjective score were (94.4±4.8) points and (93.8±5.6) points, respectively.The positive Lachman test was detected in 2 patients.No inhibition of proximal tibial epiphyseal growth was observed.Conclusions:Arthroscopic reduction with non-absorbable Krackow-" 8" suture and establishment of bone tunnels using a 2.0 mm perforated Kirschner wire for the treatment of tibial intercondylar ridge fractures in children has the advantages of simple surgical procedures, minimal invasiveness, which is an effective treatment method.