2.Cloning and Expression of MTSase and MTHase from Sulfolobus solfataricus in E.coli
Xiao-Bin CHEN ; Jian-Ping LIN ; Zhi-Hua JIN ; Pei-Lin CEN ;
Microbiology 1992;0(05):-
The genes of maltooligosyl trehalose synthase (MTSase) and maltooligosyl trehalose tetrahydrolase(MTHase) from Sulfolobus solfataricus ATCC 35092 were amplified using PCR. The expression plasmids, pTrc99a-MTSase and pTrc99a-MTHase, were constructed by inserting these two DNA fragments into E. coli expression vector pTrc99a. The specific activity of MTSase and MTHase in E. coli BL21(DE3) at optimal fermentation conditions reached 31.3U/g (wet cell) and 403U/g (wet cell), respectively. The biotransformation of partially hydrolyzed starch to trehalose catalyzed by MTSase and MTHase was carried out at 75℃ and pH 5.0. The highest yield of trehalose (ca. 53.6%) was gained when the original starch concentration was 15%(w/v) and the DE value was 10.
3.Biphasic calcium phosphate/polyvinyl alcohol scaffolds prepared by 3D-printing at room temperature and their impact on in vitro osteogenic differentiation
Yue SONG ; Kaifeng LIN ; Shu HE ; Shuaishuai ZHANG ; Bin LIU ; Long BI ; Guoxian PEI
Chinese Journal of Orthopaedic Trauma 2017;19(5):409-416
Objective To prepare biphasic calcium phosphate/polyvinyl alcohol scaffolds by 3D printing at room temperature and explore the effect of 3D scaffolds on in vitro osteogenic differentiation of the bone marrow mesenchymal stem cells (BMSCs).Methods After biphasic calcium phosphate and polyvinyl alcohol solutions were mixed,the biphasic calcium phosphate/polyvinyl alcohol composite scaffolds were prepared by room temperature 3D printing combined with freeze drying technique.Non-printing scaffolds were prepared by injection molding.The surface microstructure,porosity,elastic modulus and hydrophilicity of the 2 sorts of scaffolds were measured.The cytological experiments were carried out in 3 groups (n =3):printed scaffold group,non-printed scaffold group and blank control group (no scaffold).After the BMSCs were seeded onto the scaffolds for 7 and 14 days,the 3 groups were compared in terms of cellular proliferation,alkaline phosphatase activity and expression levels of osteogenesis-related genes.Results 3D composite scaffolds with controllable pore size and porosity were prepared successfully,with an average porosity of 59.6% ± 3.6% and an average elastic modulus of 429.3 ± 54.3 kPa.After culture for 7 and 14 days,the cellular absorbance values in the printed scaffold group (0.987 ± 0.047 and 1.497 ± 0.076) were significantly higher than those in the non-printed scaffold group (0.767 ±0.063 and 1.181 ±0.098) (P < 0.05) which were in turn significantly higher than those in the blank control group (0.532 ±0.046 and 0.895 ± 0.062) (P < 0.05).After culture for 7 and 14 days,the ALP activity and expression levels of osteogenesis-related genes in the printed and non-printed scaffold groups showed no significant between-group differences (P > 0.05),but were significantly higher than those in the blank control group (P < 0.05).Conclusions Tissue-engineered composite biphasic calcium phosphate/polyvinyl alcohol scaffolds with controllable pore size and good connectivity can be prepared by freeze-drying and room temperature 3D printing techniques.Co-culture of the scaffolds and BMSCs in vitro promotes adhesion,proliferation and osteogenic differentiation of the cells.
4.Effect of history of open nephrolithotomy on percutaneous nephrolithotomy
Huimin ZHANG ; Qiong PEI ; Haijun CUI ; Lin WANG ; Jianhui LIU ; Bin GAO ; Shengyong CAI ; Peilin CHEN
Chinese Journal of Geriatrics 2011;30(7):578-580
Objective To investigate the effects of previous open nephrolithotomy on the technical features, outcomes and morbidities of subsequent percutaneous nephrolithotomy (PCNL). Methods Ninety-eight patients who underwent PCNL from January 2006 to January 2011 were selected in this study. The 34 patients of them who had previous open nephrolithotomy on the same kidney were assigned as group A, and the other 64 patients who had no previous open surgery as group B. The data of operation time, blood transfusion quantity, residual stones rate, hospitalization time and time of tube evulsion were collected and compared between the two groups. Results There were no significant differences between the group A and B with respect to the mean operative time [(84.0±24.6) min vs. (94.0±22.7) min, t=1.372, P=0.177], hospitalization time [(6.5±1.1)days vs. (6.3±1.8)days, t=0.49, P=0.261], blood transfusion quantity [(82.9±10.6) ml vs. (85.0±11.8) ml, t=0.415, P=0.682], kidney and colostomy channels [single channel(70.6% vs. 75.0%), double channel (29.4% vs. 25.0%), χ2 =0.22, P=0.638] and residual stones rate (5% vs. 3%,χ2=0.42, P=0.282). Conclusions When PCNL is performed after previous open nephrolithotomy, there is no difference in success rate and morbidities.
5.In vivo kinematics analysis of the knee joint of people after fixed-bearing posterior stabilized total knee arthroplasty
Xiaojun SHI ; Jiangli LIN ; Bin SHEN ; Jing YANG ; Zongke ZHOU ; Pengde KANG ; Fuxing PEI
Chinese Journal of Orthopaedics 2013;(3):259-265
Objective To compare the kinematics of the knee joint of normal Chinese people and people after fixed-bearing posterior stabilized total knee arthroplasty while doing weight-bearing deep knee-bending using fluoroscopy analysis.Methods From June to December 2010,ten volunteers and ten patients with fixed-bearing posterior stabilized total knee arthroplasty (TKA) were required to perform weight-bearing deep knee-bending activity under surveillance of roentgenization,motion between femur and tibia was analyzed with interval of 15° according to two-to three-dimensional (2D-3D) registration technique.Results During weight-bearing deep knee-bending,the average weight-bearing maximal flexion was 136° in the normal group,which was significantly higher than the 125° in the TKA group.All 10 normal knees present posterior translation of femoral condyle during deep knee-bending,posterior translation of the lateral condyle was greater than medial condyle,thereby creating a medial pivot type of axial rotational pattern in which the tibia internally rotates relative to the femur as flexion increased.The average amount of posterior femoral translation of the medial condyle was 7.3±1.2 mm,whereas the lateral condyle translated posteriorly 19.3±3.1 mm.All knees experienced tibia rotated internally during progressive flexion,and the average amount of axial rotation for the ten subjects from 0° to 135° was 23.8±3.4°.From extension to maximal flexion,the average amount of posterior translation of medial condyle was 1.4±0.6 rmm,whereas the lateral condyle translated 6.4±1.7 mm in the posterior direction in the fixed-bearing posterior-stabilized TKA.The average amount of tibial internal rotation was 8.5°±3.4°.Conclusion In normal Chinese people,during knee flexion activities the lateral condyle experiences significantly more amount of posterior translation than the medial condyle,leading to the tibia present medial pivot internal rotation relative to the femur,and the PFC-Sigrna fixed-bearing posterior stabilized designs has the similar kinematics with normal knee,but the posterior condylar translation and tibial axial rotation are much less than the normal.
6.Solanine induces the apoptosis of human prostate cancer cells via ROS/p38 signaling pathway
Weifeng ZHONG ; Nanhui CHEN ; Yuqing HUANG ; Pei WAN ; Yifeng LIN ; Huiming JIANG ; Kaihua ZHONG ; Bin PAN ; Siping LIU
Journal of Xi'an Jiaotong University(Medical Sciences) 2017;38(3):457-461,封3
Objective To investigate the molecular mechanism of solanine-induced apoptosis of prostate cancer cells Du145 and LNCaP.Methods The effects of solanine on the viability of Du145 and LNCaP cells were evaluated by MTT assay.The generation of intracellular reactive oxygen species (ROS) and solanine-induced apoptosis were measured by flow cytometry.The protein levels of p38 and p-p38 expressions were examined by Western blot.Results Solanine significantly inhibited the viability of Du145 and LNCaP cells in a dose-dependent manner (P<0.01).The inhibition of solanine on cell viability was suppressed by the ROS scavenger NAC.ROS generation,apoptosis and phosphorylation of p38 were induced by treatment with solanine at 40 μmol/L for 24 h.The expression of p38 and solanine-induced apoptosis were suppressed by NAC and SB203580.Conclusion Solanine induces the apoptosis of human prostate cancer cell via the RO.S-p38 signaling pathway.
7.Study on the effect of treatment and the status of drug resistance among HIV/AIDS patients treated with highly active antiretroviral therapy in Shandong province.
Jing ZHANG ; Bin LIN ; Lin LIN ; Pei-ju YANG ; Ji-hua FU ; Han-ping LI ; Jing-yun LI
Chinese Journal of Epidemiology 2007;28(11):1108-1111
OBJECTIVETo understand the effect of highly active antiretroviral therapy (HAART) on AIDS patients, and to explore the prevalence and the impact of HIV-1 drug resistance in Shandong province.
METHODS2 cross-sectional studies were carried on in 2004 and 2005, to collect data on clinical symptoms and compliance of the AIDS patients with HAART through questionnaire. Informed-consent principle was followed to test on immunological, viral and laboratory index of them. HIV-1 drug genotype resistance by sequencing the gene of HIV-POL after RT-PCR was performed and analyzed.
RESULTS31 AIDS cases with and. 27 AIDS cases without HAART, were studied. 83.3% and 64.5% of the AIDS patients with HAART showed that the CD4+ T cell count was rising to over 350/microl, in the first study (2004) and in the second (2005) study respectively but still 45.8% and 45.2% of AIDS patients under HAART in the 2 years showed a decreasing HIV load under the detected limit. However, these findings were showing remarkable difference when compared with the AIDS without HAART. 7 drug resistance gene sites were found in AIDS patients with HAART and in AIDS patients without HAART. The rate on high degree drug resistance mutation and total drug resistance rate of mutation of the former were higher remarkably than those of the latter.
CONCLUSIONMost of the AIDS patients with HAART met the purpose of rebuilding immunity and control of HIV,as well as alleviation of symptoms. Although the drug resistance stain appeared in Shandong,but had little effect on HAART. AIDS; Drug resistance; Highly active antiretroviral therapy
Acquired Immunodeficiency Syndrome ; drug therapy ; Adult ; Antiretroviral Therapy, Highly Active ; CD4 Lymphocyte Count ; China ; Cross-Sectional Studies ; Drug Resistance, Viral ; genetics ; Genotype ; HIV-1 ; drug effects ; genetics ; Humans ; Middle Aged ; Patient Compliance ; Prevalence ; Surveys and Questionnaires ; Viral Load
9.Role of bcl-2 transcriptional regulation induced by calmodulin I pressure overload rat hypertrophic hearts.
Qi ZHOU ; Ying-Bin XIAO ; Jian LIU ; Pei-Yong WANG ; Lin CHEN ; Qian-Jin ZHONG ; Xue-Feng WANG
Acta Physiologica Sinica 2005;57(6):731-736
This study was designed to evaluate the role of bcl-2 transcriptional regulation induced by calmodulin I (CaM I) in pressure overload rat hypertrophic hearts. The model of hypertensive Sprague-Dawley rat was established by abdominal aortic constriction. The hearts were collected four weeks after abdominal aortic constriction. Velocity and isopyknic gradient centrifugation was employed to fractionate rat myocardial nuclei. Western blot analysis revealed a marked increase in phosphorylated cAMP response-element binding protein (pCREB) of cardiac hypertrophy group compared with that in control group (P<0.05), while the protein level of cAMP response-element binding protein (CREB) was constant (P>0.05). Immunohistochemistry results showed a significant increase of CaM I protein in cardiac hypertrophy group relative to the control group (P<0.05). Nuclear run off transcription assay displayed a significant increase in bcl-2 mRNA treated with trifluoperazne compared with non-drug treatment (P<0.05). The results obtained suggest that the transcription of bcl-2 is possibly regulated by CaM I hypertrophic rat hearts, and CREB phosphorylation seems to be a minor factor in bcl-2 transcriptional regulation.
Animals
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Aorta, Abdominal
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pathology
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Calmodulin
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physiology
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Cardiomegaly
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metabolism
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physiopathology
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Constriction
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Cyclic AMP Response Element-Binding Protein
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genetics
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metabolism
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Male
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Phosphorylation
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Proto-Oncogene Proteins c-bcl-2
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genetics
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metabolism
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RNA, Messenger
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genetics
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metabolism
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Rats
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Rats, Sprague-Dawley
10.Effects of mixed-tocopherols and eicosapentaenoic acid on oxidized LDL-induced oxidative damage and inflammatory cytokine secretion in human umbilical vein endothelial cells.
Ming-Bin XIE ; Mei-Lin LIU ; Ying-Shuo HUANG ; Pei LI ; Ya-Bing YANG ; Xia-Huan CHEN
Chinese Journal of Cardiology 2011;39(3):242-246
OBJECTIVETo observe the influence of either alone or combined mixed-tocopherols combined with eicosapentaenoic acid (EPA) and α-Tocopherol use on oxidized LDL (oxLDL) induced 8-hydroxy-2'-deoxyguanosine (8-OHDG) and interleukin-6 (IL-6) secretion by human umbilical vein endothelial cells (HUVECs) and to explore the potential mechanism.
METHODCultured HUVECs in vitro were incubated with oxLDL, oxLDL + α-tocopherol, oxLDL + mixed-tocopherols, oxLDL + EPA, oxLDL + α-tocopherol + EPA, oxLDL + mixed-tocopherols + EPA for 24 hours, respectively. Secretion of 8-OHDG and IL-6 were detected by cell enzyme linked immunosorbent assay (ELISA). The expressions of superoxide dismutase (SOD), protein kinase C-δ (PKC-δ), phosphorylated PKC-δ (p-PKC-δ) were analyzed by Western blot.
RESULTS8-OHDG and IL-6 secretion of HUVECs was significantly increased significantly after incubated with oxLDL for 24 hours which could be significantly attenuated in the presence of tocopherols and EPA (alone or in combination, all P < 0.05) while the strongest inhibition effects were seen with combined use of mixed-tocopherols and EPA. Moreover, combination of mixed-tocopherols and EPA could also significantly increase SOD activity and decrease PKC activity (all P < 0.05). However, the protein expression of SOD and PKC-was similar among groups.
CONCLUSIONCombined mixed-tocopherols + EPA use enhanced the inhibiting effects on the secretion of 8-OHDG and IL-6 in oxLDL stimulated HUVECs which might be linked with increased SOD activity and reduced p-PKC activity.
Antioxidants ; Cells, Cultured ; Deoxyguanosine ; analogs & derivatives ; secretion ; Eicosapentaenoic Acid ; pharmacology ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; Humans ; Interleukin-6 ; secretion ; Lipoproteins, LDL ; adverse effects ; Protein Kinase C ; metabolism ; Superoxide Dismutase ; metabolism ; alpha-Tocopherol ; pharmacology