2.Autophagy and cardiocyte apoptosis after heterotopic transplantation of the mouse heart preserved in high-pressured mixed gas
Rui ZHANG ; Shuai HUANG ; Qi FU ; Shaoyi ZHENG ; Huiming GUO ; Jimei CHEN ; Jian ZHUANG ; Ping ZHU
Journal of Medical Postgraduates 2015;(12):1236-1241
Objective Heart transplantation is an effective treatment of end-stage heart diseases and extending the time of donor heart preservation helps to make up for the shortage of donor hearts. This study was to investigate whether high-pressured mixed gas ( HPMG) of carbon monoxide and oxygen could prolong the time of donor heart preservation and its mechanisms. Methods Forty-eight C57BL/6 male mice aged 4-6 weeks were randomly divided in-to four groups of equal number:control ( the donor heart isolated but not transplanted) , immediate transplantation ( the donor heart transplanted right after isolated) , HTK-preservation ( the donor heart preserved in histidine-tryptophan-ketoglutarate solution for 24 hours after isolated, and HPMG preservation ( the donor heart preserved in an HPMG chamber with the oxygen partial pressure of 3200 hPa and carbon monoxide partial pressure of 800 hPa for 24 hours after isolated) .Another 36 recipient mice aged 6-8 weeks were randomly assigned to receive the donor heart immediately after harvested (n=12), preserved in HTK solution (n=12), or preserved in HPMG (n=12).At 2 hours after transplantation, the status of heart re-beating and cardiac function were compared among different groups of recipient mice.At 24 hours, tissues were taken from the transplanted hearts for examination of pathologic changes by HE stai-ning, detection of the apoptosis of cardiac cells by TUNEL, and determination of the expressions of microtubule-associated protein 1 light chain 3 -Ⅱ(LC3-Ⅱ) and B cell lymphoma/leukemia-2 (Bcl-2) by Western blot. Resul ts The re-beating rates of the imme-diately transplanted and HPMG-preserved hearts were significantly higher than that of the HTK-preserved ones (P<0.05).At 2 hours after transplantation, the cardiac function scores were 2.5 (2.0-2.9), 0.8 (0.5-1.0), and 4.5 (4.0-4.5) in the immediate implantation, HPMG-preservation and HTK-preservation groups respectively, with statistically significant differences between any two groups (P<0.05).The expressions of LC3-Ⅱand Bcl-2 were 2.06 ±0.29 and 0.87 ±0.18 in the HPMG-preserved heart recipients and 1.24 ±0.20 and 2.07 ±0.32 in the immediately transplanted heart recipients, both higher than 0.13 ±0.03 and 0.19 ±0.02 in the controls and 0.16 ±0.06 and 0.26 ±0.08 in the HTK-preserved heart recipients (P<0.05), the Bcl-2 higher in the HTK-pre-served heart recipients than in the controls (P<0.05), and the LC3-Ⅱ expression higher in the HPMG-preserved heart recipients than in the immediately transplanted heart recipients (P<0.05).HE staining showed that cell edema and inflammatory cell infiltration were more obvious in the HPMG-preserved heart recipients than in the controls and immediately transplanted heart recipients but less obvious than in the HTK-preserved heart recipients.The rate of cell apoptosis was dramatically increased in the HPMG-and HTK-pre-served heart recipients ([5.04 ±1.77]%and [26.72 ±5.23]%) in comparison with the controls ([1.08 ±0.56]%) (P<0.01) and immediately transplanted heart recipients ([2.13 ±1.71]%) (P<0.01) but decreased in the HPMG as compared with the HTK-preserved heart recipients (P<0.01). Conclusion High-pressured mixed gas preservation can reduce cold ischemia-reperfu-sion injury of the donor heart, which may be associated with its promotion of autophagy, provision of energy to cells, and apoptosis of cardiocytes in the donor heart.
3.Quality assurance of clinical biochemistry testing:a mualti-center study based reference interval for clinical chemistry tests in the Chinese population
Chuanbao ZHANG ; Xianzhang HUANG ; Lanlan WANG ; Runqing MU ; Baishen PAN ; Jie ZHANG ; Wenxiang CHEN ; Junha ZHUANG ; Hengjian HUANG ; Yueyun MA ; Xiaoou YU ; Wei GUO ; Rui QIAO ; Hong SHANG
Chinese Journal of Laboratory Medicine 2015;(5):301-305
Objective To verify and monitor the performance of accuracy, precision and comparability of 26 clinical biochemical analytes (29 methods) in the six centers involved in multi-centers reference intervals research, and to ensure the reliability of theirmeasurement results.Methods During the period of the systems evaluating, two levels of commercial quality control materials and fresh frozen human serum reference materials were applied to verify the performance of inter-laboratory precision and accuracy of analysis systems. During the period of samples testing, the commercial quality control materials were measured whenever samples were analysed, the fresh frozen serum reference materials were measured once a month.The coefficient of variations (CVs), bias and total errors were calculated to assess the precision, accuracy and comparability.Results Verification of precision and accuracy: ( 1 ) the ranges of CVs of 29 methods in the six laboratory laboratories were 0.4%-6.0%, the CVs of all 29 methods met the criterion . (2) The overall average bias of the analysis systems of 21 analytes (24 methods) ranged from -5.15%( ALT) to 4.46% ( Ur ) .Among 24 methods the overall average bias of TP, Glu-GOD, Ur, Cl, Ca exceeded the acceptable range.The quality assessment during the period of samples testing:(1) The overall average bias ranged from -1.95%(Ca) to 2.92%(Ur), median 1.26%, they all met the requirements of relevant standards.( 2 ) When commercial control materials were tested, the requirements of CVs were fulfilled for most methods in the six laboratories,and the CVs of TP, Alb, Cl, Ca exceeded the acceptable range.The overall average TE of all methods met the quality specification for the C-N controls material.For the C-P control material, only the overall average TE of TP (5.05%) exceeded thearceptable range while the other methods met the requirement in criterion.Conclusions The performance of precision and accuracy of the analysis systems used in the six laboratories passed the verification.During the period of sample testing, the performance of precision and accuracy of the most methods in the 6 laboratories met the requirements of quality specifications, and the overall performance was good.Because of the limitation of current technology the performance of some methods didn't fulfill the requirement of specifications, and need to be improved.
4.Construction and preparation of human metapneumovirus vaccine based on influenza virus vector
Mengxue GAO ; Xiaoman LIU ; Liru GUO ; Mei KONG ; Zhichao ZHUANG ; Aiping YU ; Rui LI ; Xiaoyan LI
Chinese Journal of Experimental and Clinical Virology 2024;38(1):77-85
Objective:To construct and prepare recombinant virus strains chimeric with human metapneumovirus (HMPV) antigenic epitopes.Methods:Recombinant influenza virus vectors which chimeric with different HMPV antigenic epitopes were rescued by reverse genetics using eight-plasmid system. The recombinant influenza virus strain used the internal genes of A/PR/8/34 (PB1, PB2, PA, NP, NS, M, HA, and NA) as a backbone, with concomitant genetic modifications to insert the B-cell epitopes of HMPV into the HA gene, and the CTL+ Th cell epitopes of HMPV into the NA gene. Preparation of recombinant influenza virus strains using reverse genetics in a " 7+ 1" model. The recombinant virus strains were evaluated by measuring hemagglutinin (HA) titers, half tissue culture infectious dose (TCID 50) and growth curves. Sequencing analysis was conducted to verify whether the rescued viruses carried the chimeric HMPV epitopes. Results:The epitopes of HMPV were inserted into the influenza virus genome and two recombinant influenza virus strains were rescued successfully, named as FLU/HMPV/B and FLU/HMPV/CTL+ Th. HA titers of the recombinant strains were both 2 7, their TCID 50 were 10 5.2/ml and 10 5.0/ml, respectively. After cultured for three passages in chick embryo, these two recombinant strains could proliferate steadily. Whole genome sequencing verified that the FLU/HMPV/B carried the B-cell epitopes of HMPV, the FLU/HMPV/CTL+ Th carried the CTL and Th cell epitopes of HMPV. Growth curve tests also verified that the recombinant strains could proliferate steadily in eggs. Conclusions:Two recombinant influenza virus vector strains carrying the B cell, CTL and Th epitopes of HMPV were rescued successfully. The result of the recombinant virus strains in terms of growth characteristics as well as genetic stability indicate that they meet the requirements for proceeding to the next step of animal experiments. The immunogenicity and immunoprotective effect will be further evaluated by mouse experiments. Ultimately new ideas for the realization of " one vaccine for two uses" or " one vaccine formultiple uses", as well as a new strategy for the development of HMPV vaccine will be proposed.
5.Estrogen on stimulating the angiogenesis of children's hemangioma in vitro.
Cheng-hong JIANG ; Fu-lian ZHUANG ; Ba-rui HUANG ; Zhi-hui GUO ; Yi-de XIE ; Ya-kuan ZHOU ; Biao WANG
Chinese Journal of Plastic Surgery 2007;23(2):86-90
OBJECTIVETo demonstrate that estrogen stimulates the angiogenesis of children' s hemangioma.
METHODSA piece of hemangioma biopsy was embedded in fibrin gel, and a model in vitro of angiogenesis of human hemangioma was then established. The angiogenesis of hemangioma in each group was interfered by the estrogen and tamoxifen. There were four groups divided into the followings: the group with estrogen, the group with tamoxifen, the group with estrogen + tamoxifen and the control. The dimension of newborn tubule area in the 3rd, 6th, 9th day after the culture was calculated to compare statistically differences among the groups.
RESULTSIn the model of angiogenesis of hemangioma, microvessels grew out from the tissue sample in 2 to 3 days after the culture, and in 8 to 9 days a complex network of microvessels had been shown, the tending to inactivity. On the 3rd,6th and 9th day after the culture the dimension of newborn tubule area of the group of estrogen [(2.84 +/- 0.20) mm2 (12.93 +/- 0.85) mm2 (22.47 +/- 1.40) mm2] were larger than those of the control [(1.98 +/- 0.17) mm2, (7.51 +/- 0.48) mm2, (11.26 +/- 0.73) mm2]. Those of the group of estrogen + tamoxifen [(1.08 +/- 0.11) mm2, (3.54 +/- 0.31) mm2, (5.72 +/- 0.40 mm2] and the group of tamoxifen [(1.13 +/- 0.14) mm2 (4.26 +/- 0.29) mm2, (6.08 +/- 0.42) mm2] were smaller than those of the groups of the estrogen and the control (P < 0.05).
CONCLUSIONSThe estrogen may stimulate the angiogenesis of children's hemangioma, and the tamoxifen may reverse the process.
Child ; Estrogens ; adverse effects ; Hemangioma ; pathology ; Humans ; In Vitro Techniques ; Neovascularization, Pathologic ; pathology
6.A multicenter study of the effectiveness of interferon alpha-1b (Hapgen) in treating HBeAg positive chronic hepatitis B patients.
Rui JIN ; Xin-hui GUO ; Jian-feng HUANG ; Zhuang LIU ; Han-wei LI ; Zhen-biao WANG ; Bin ZHANG ; Yu-min XIE
Chinese Journal of Hepatology 2011;19(1):25-28
To compare the efficacy and safety of interferon a-1b and interferon a-1b combined with lamivudine in the treatment of HBeAg positive chronic hepatitis B (CHB), to analyze the impact of variable factors on the efficacy, and to investigate the individualized anti-viral regimen for CHB patients. 111 CHB patients were enrolled and randomly divided into two groups. Group A: patients received interferon a-1b (49 patients, 50mug I. M. , qod. ) , Group B: interferon a-1b (idem) combined with lamivudine for 6-12 months or longer(62 patients, 100 mg, P.O. , q.d. ). (1) The HBeAg seroconversion rates of treatment by 12 and 18 months were 28.6% and 36.7% in group A, 29.0% and 38.7% in group B, respectively, no significant difference found between the two groups at the end of treatment (x2=0.003, P value is more than 0.05; x2=1.500, P value is more than 0.05). (2) The HBV DNA undetectable rates of treatment by 6 months, 12 months and 18 months were 8.2%, 53.1% and 57.1% in group A, 66.1%, 83.9% and 88.7% in group B, respectively, still no significant difference existed between the two groups (x2=38.150, P value is less than 0.05; x2=12.073, P value is less than 0.05, x2=14.459, P value is less than 0.05). (3) In group A, the HBeAg seroconversion rates for male and female patients were 34.5% and 40.0% respectively, no significant difference found between. As regard ages the rates were 34.9% and 50.0% for patients younger or more than 40 years of age, no significant difference existed between. The HBeAg seroconversion rate was higher in patients with lower baseline serum HBV DNA loads ( less than 6 log10 copies/ml) . (4) The rates of patients with fever and blood abnormality were 36.7% and 34.7% in group A, 32.3% and 27.4% in group B, respectively. The total incidences of adverse events were similar between group A and B (x2=0.244, P value is more than 0.05; x2=0.682, P value is more than 0.05). (5) The ratio of drug resistance in group B was only 1.6%. The adverse events of interferon a-1b treatment for CHB are low and mild. The HBeAg seroconversion rate persistently raises with the extension of interferon a-1b treatment course. The HBV DNA undetectable rate of interferon a-1b combined with lamivudine is significantly higher than that of interferon a-1b and the drug resistance of lamivudine can be reduced obviously by combination therapy.
Adult
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Antiviral Agents
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therapeutic use
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Drug Therapy, Combination
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Female
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Hepatitis B e Antigens
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blood
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Hepatitis B, Chronic
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blood
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drug therapy
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Humans
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Interferon-alpha
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administration & dosage
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therapeutic use
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Lamivudine
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therapeutic use
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Male
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Middle Aged
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Young Adult
7.A three-dimension in vitro model for angiogenesis of hemangioma.
Cheng-Hong JIANG ; Fu-Lian ZHUANG ; Ba-Rui HUANG ; Zhi-Hui GUO ; Yi-De XIE ; Ya-Kuan ZHOU ; Biao WANG ; Wen-lie CHEN
Chinese Journal of Plastic Surgery 2005;21(5):364-367
OBJECTIVETo create a three dimension (3D) in vitro model for angiogenesis of hemangioma.
METHODThe fragment of hemangioma specimen was embedded in fibrin gel to set up the three-dimension (3D) in vitro model for angiogenesis of hemangioma.
RESULTIn the model, microvessels grew out from the tissue fragments at the 2nd to 3rd day after culture, and at the 8th to 9th day a compact network of microvessels come into being, then tending to be stationary. The compact network around the tissue fragment was confirmed to be blood vessels by immunohistochemistry and electron microscopy.
CONCLUSIONThis model helps to study the mechanism of hemangioma angiogenesis and investigate the drugs of anti-angiogenesis.
Cells, Cultured ; Endothelium, Vascular ; Hemangioma ; Humans ; Models, Cardiovascular ; Neovascularization, Pathologic
8.Effects of erythropoietin on neuronal proliferation and apoptosis in neonatal rats after infection-induced brain injury.
Fa-Lin XU ; Jia-Jia GUO ; Fang-Li ZHUANG ; Yan-Hua ZHANG ; Cai-Hong WANG ; Rui AN
Chinese Journal of Contemporary Pediatrics 2014;16(3):290-294
OBJECTIVETo investigate the effects of erythropoietin (EPO) on the neuronal proliferation and apoptosis in neonatal rats after infection-induced brain injury and the neuroprotective mechanism of EPO in neonatal rats with infection-induced brain injury.
METHODSTwenty-six two-day-old neonatal rats were randomly divided into 3 groups: control group (intraperitoneally given an equal volume of normal saline), lipopolysaccharide (LPS) group (intraperitoneally given LPS 0.6 mg/kg), and EPO group (intraperitoneally given LPS 0.6 mg/kg and EPO 5 000 U/kg). These groups were injected with respective drugs for 5 consecutive days. Meanwhile, each group was intraperitoneally injected with 5-bromo-2'-deoxyuridine (BrdU) (50 mg/kg) once a day for 5 consecutive days. The expression of BrdU and cleaved Caspase-3 in the hippocampal dentate gyrus was detected by immunohistochemistry at 24 hours after the last injection.
RESULTSThe number of neuronal cells in the hippocampal dentate gyrus in the LPS and EPO groups was significantly greater than in the control group (P<0.05), but there was no significant difference between the LPS and EPO groups. The EPO group had a significantly higher number of BrdU-positive cells in the subgranular zone of hippocampal dentate gyrus than the LPS group (51±9 vs 29±6; P<0.05), but a significantly lower number of BrdU-positive cells than the control group (51±9 vs 67±12; P<0.05). The EPO group had a significantly lower number of cleaved Caspase-3-positive cells in the subgranular zone of hippocampal dentate gyrus than the LPS group (27.9±1.5 vs 34.0±1.3; P<0.05), but a significantly higher number of cleaved Caspase-3-positive cells than the control group (27.9±1.5 vs 21.0±1.7; P<0.05).
CONCLUSIONSEPO can promote hippocampal neuronal proliferation and reduce neuronal apoptosis in neonatal rats after infection-induced brain injury.
Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Brain Diseases ; drug therapy ; pathology ; Bromodeoxyuridine ; metabolism ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Erythropoietin ; pharmacology ; therapeutic use ; Hippocampus ; pathology ; Neurons ; pathology ; Rats ; Rats, Sprague-Dawley
9.Identification of Differentially-expressed Genes in Intestinal Gastric Cancer by Microarray Analysis
Zang SHIZHU ; Guo RUIFANG ; Xing RUI ; Zhang LIANG ; Li WENMEI ; Zhao MIN ; Fang JINGYUAN ; Hu FULIAN ; Kang BIN ; Ren YONGHONG ; Zhuang YONGLONG ; Liu SIQI ; Wang RONG ; Li XIANGHONG ; Yu YINGYAN ; Cheng JING ; Lu YOUYONG
Genomics, Proteomics & Bioinformatics 2014;(6):276-284
Gastric cancer (GC) is one of the most frequent malignant tumors. In order to systematically characterize the cellular and molecular mechanisms of intestinal GC development, in this study, we used 22 K oligonucleotide microarrays and bioinformatics analysis to evaluate the gene expression profiles of GC in 45 tissue samples, including 20 intestinal GC tissue samples,20 normal appearing tissues (NATs) adjacent to tumors and 5 noncancerous gastric mucosa tissue samples. These profiles allowed us to explore the transcriptional characteristics of GC and determine the change patterns in gene expression that may be of clinical significance. 1519 and 1255 differentially- expressed genes (DEGs) were identified in intestinal GC tissues and NATs, respectively, as determined by Bayesian analysis (P < 0.001). These genes were associated with diverse functions such as mucosa secretion, metabolism, proliferation, signaling and development, which occur at different stages of GC development.
10.Clinical value of high frequency ultrasound in the diagnosis of hip joint involvement in patients with ankylosing spondylitis
Guo-Yan XUE ; Ying-Qi LI ; Rui WANG ; Wen-Xing WANG ; Zhuang DONG ; Jian-Feng DING
China Journal of Orthopaedics and Traumatology 2024;37(4):374-380
Objective To analyze the correlation between hip joint musculoskeletal ultrasound score and ankylosing spondylitis(AS)disease activity,as well as to investigate the value of high frequency ultrasound in the assessment of hip joint involvement in AS.Methods The clinical data of 244 patients with AS who were treated in the rheumatology department of from March 2019 to March 2022 were retrospectively analyzed.Among them,there 174 males and 70 females,aged from 19 to 58 years old with an average of(34.22±9.49)years old;the disease duration of AS patients ranged from 8 months to 26 years,with an average of(13.68±4.04)years.The 244 patients were divided into disease group(83 cases)and control group(161 cases)based in the presence of hip joint involuement.According to the the disease activity,patients in the disease group were further categorezed into active phase(45 cases)and stable phase(38 cases).The ultrasound scores of patients in the active and stable phases of the disease group and the control group were compared.Relevant factors of hip joint involvement in AS patients were analyzed,and analyze the correlation between ultrasound score and Bath ankylosing spondylitis disease activity score index(BASD AI),Bath ankylosing spondylitis functional index(BASFI),visual analogue score of pain(VAS),C-reactive protein(CRP),erythrocyte sedimentation rate(ESR),and the correlation between hip joint capsule score and tendon attach-ment end score and BASDAI,BASFI,VAS,CRP and ESR.Results The hip joint capsule score(3.06±1.12),femoral head score(1.45±0.43),tendon attachment end score(3.28±1.30)and total ultrasound score(6.65±2.31)of the disease group were higher than those of the control group(1.51±0.48)、(0.66±0.27)、(1.61±0.53)、(3.81±1.44)scores(P<0.05).Multifactor Logstic re-gression analysis showed that the course of disease,hip joint capsule score and total ultrasound score were independent risk factors for hip involvement in AS patients.The hip capsule score(3.65±1.22)and total ultrasound score(8.28±2.33)in the ac-tive phase of the disease group were higher than those in the stable phase(2.48±1.04)、(6.82±1.96)scores(P<0.05).The hip joint capsule score and total ultrasonic score of AS patients were positively correlated with BASDAI,BASFI,VAS,CRP,and ESR(P<0.05,P<0.01).The score of tendon attachment end was positively correlated with CRP(P<0.05).The score of joint capsule effusion in AS patients was positively correlated with BASDAI,BASFI and VAS(P<0.05,P<0.01).The synovial blood flow score was positively correlated with BASDAI,VAS,CRP and ESR(P<0.05,P<0.01).The synovial thickening score was positively correlated with BASDAI,BASFI,VAS,CRP and ESR(P<0.05,P<0.01).There was no correlation between the score of tendon attachment end and BASDAI,BASFI,VAS,CRP and ESR.Conclusion There is a correlation between hip joint ul-trasonic score of hip joint and clinical indexes in AS patients.Hip joint capsule score and total ultrasonic score were indepen-dent risk factors for hip involvement in AS patients.High frequency ultrasound exhibits clinical value in the diagnosis of hip joint involvement in AS patients.