2.In vitro biocompatibility of polyhydroxybutyrate-co-volerate membrane with human bone marrow mesenchymal stem cells
Tao ZHANG ; Zhixu HE ; Chuan YE ; Jielin LIU ; Minxian MA ; Bo SUN ; Mei WANG ; Hua YANG
Chinese Journal of Tissue Engineering Research 2014;(12):1920-1925
BACKGROUND:Polyhydroxybutyrate-co-volerate (PHBV) is a noticeable tissue engineering material of polyhydroxyalkanoates family. It has the properties of low immune rejection response and good biocompatibility, and its degradation products are non-toxic.
OBJECTIVE:To investigate the biocompatibility of PHBV membrane material and human bone marrow mesenchymal stem cel s in vitro.
METHODS:Human bone marrow mesenchymal stem cel s at passage 3 were seeded upon PHBV membrane as experimental group and upon conventional culture plates as control group. Then we calculated the adherent cel number of two groups at 1, 2 and 4 hours and got the cel adherent rate. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide assay was used at days 2, 4, 6, 8 to observe the cel proliferation of two groups. Fluorimetric method with the fluorescent dye Hoechst 33258 was used to detect the DNA content of cel s at days 3, 6, 9 and 12 in both groups. After cel s were seeded upon PHBV membrane for 5 days, the cel growth upon the material was examined under a scanning electron microscope.
RESULTS AND CONCLUSION:When the cel s were cultured for 1 hour, the adherent rate in the experimental group was lower than that in the control group;but there were no significant differences between two groups at the other two periods. No difference was found in the cel proliferation and the DNA content between the two groups. Human bone marrow mesenchymal stem cel s seeded upon PHBV membrane for 5 days grew wel with spindle morphology and the intercel ular connections were tight and more extracel ular matrices were observed by scanning electron microscopy. Taken together, PHBV membrane material shows a good biocompatibility with human bone marrow mesenchymal stem cel s.
3.Role of MafA gene in insulin production-Analysis of heterozygous knockout mice
Chuan ZHANG ; Bo LI ; Yan CHENG ; Yuhang YAO ; Lijuan SUN ; Satoru TAKAHASHI
Chinese Journal of Endocrinology and Metabolism 2013;29(12):1048-1051
Objective To clarify the role of MafA gene in development of MODY (maturity onset diabetes of the young) by studying insulin production,gene expression,and serum glucose level in heterozygous MafA gene knockout mice.Methods C57BL/6J mice were used as control animals,MafA gene heterozygous mice were analyzed.The distribution curve of blood sugar levels over time and serum insulin of heterozygous mice were determined by using IPGTT.The sensitivity of the mice to insulin was examined by injecting insulin assay.The expression levels of genes of MafA,insulin,pdX1,Beta2,and other genes of heterozygous mice were analyzed by semi-quantitative RT-PCR.Morphological changes in pancreatic tissue and α-and β-cell counts were obtained by using immunofluorescence/histological examination.Results (1) Two weeks after birth,MafA gene heterozygous mice began to show that the blood glucose level was increased,weight was reduced,and the amount of insulin secretion was clearly decreased (P<0.05 or P<0.01) while the insulin sensitivity did not change significantly.(2)The islet volume in MafA gene heterozygous mice was increased significantly as compared with the control group.However there were no significant changes in the number of pancreatic cells,distribution patterns,and the ratio of α and β cell.(3) Semi-quantitative RT-PCR detection showed that,compared with the control group,MafA gene level,the amount of insulin and Beta2 gene in MafA gene heterozygous mice were significantly reduced(all P<0.05),while no changes in the amount of glucagons and level of Pdx1 were found.Conclusions The blood glucose level of MafA gene heterozygous mice was raised early after birth.MafA gene is likely to be a new disease ralated gene of MODY.
4.MRI study on the cortical thickness of occipital lobe in children with ametropic amblyopia
Hanjian DU ; Jian WANG ; Chuan LI ; Jiuquan ZHANG ; Li CHEN ; Bo LIU
Chinese Journal of Radiology 2008;42(1):47-49
Objective To study cortical thickness of the occipital lobe in children with ametropic amblyopia by using MRI technique and the FreeSurfer software.MethOds Nine children with ametropic amblyopia were included in the amblyopic group and 8 normal children were included in the control group.All the children underwent brain MRI on the Siemens Avanto 1.5 T scanner.For the cortical thickness analysis,3-demensional MPRAGE images were collected and analyzed with FreeSurfer software package.Cortical thickness of related regions in the occipital lobe (including the cuneus,later occipital,lingual,and pericalcarine gyri) were recorded and compared. Results The cortical thickness of the lingual,pericalcarine gyri on the left hemisphere and the cuneus,lateraloccipital,lingual gyri on the right hemisphere in amblyopic group were lower than the control group (P<0.05). Conclusion Morphological changes existed in the occipital lobe in ametropic amblyopic children.The analysis technique with the FreeSurfer package has a potential value in the clinical application.
5.Cloning of human uracil N-glycosylase and its detection in cancer tissues by quantitative RT-PCR.
Hong-Bo BAO ; Chuan-Bao ZHANG ; Jin-Fang WANG ; Chuan-Nong ZHOU ; Fang LIU ; Xiao-Hang ZHAO ; Shi-Jun QIAN
Chinese Journal of Biotechnology 2003;19(5):561-565
The uracil in DNA comes from either the misincorporation of dUTP in place of dTTP or deamination of cytosine. In the latter case, it can result in a GC to AT transition mutation if the uracil is not removed before DNA replication. Base excision repair (BER) is a major pathway for removing DNA lesions arising from endogenous processes as well as those induced by exposure to exogenous chemicals or irradiation. BER is initiated by DNA glycosylases that excise aberrant bases from DNA by cleavage of the N-glycosidic bond linking to the base of its deoxyribose sugar. Uracil N-glycosylase (UNG) is the enzyme responsible for the first step in the BER pathway that specifically removes uracil from DNA. The UNG gene undergoes both temporal and spatial regulation mainly at the level of transcription. Normally cancer cells undergo over-proliferation and up-regulate their UNG during tumorigenesis. In this study we examine the correlation between UNG level and carcinogenesis, and explore the possibility of using UNG as a marker for cancer diagnosis. Human UNG gene was amplified from the total RNA of the human choriocarcinoma cell line, JEG-3, by RT-PCR. After purification, the 942bp full-length UNG cDNA coding sequence was digested with EcoR I and Sal I, and cloned into the digested pET-21 to construct a recombinant vector, pUNG. The UNG protein was expressed under the control of T7 promoter in E. coli BL21 (DE3) cells induced with IPTG. After ultrasonic treatment, the cell lysate and precipitate were analyzed by SDS-PAGE and a 39kD band was detected. The plasmid was serially diluted at appropriate concentrations and employed as standards in the subsequent quantification. Total RNAs were extracted from 18 pairs of clinical samples, each pair contains a sample of esophageal squamous cell carcinoma (ESCC) tissue and its surrounding normal esophageal epithelia. The copy numbers of UNG mRNA in these RNA samples were determined by real-time quantitative RT-PCR using a Lightcycler (Roche). UNG was present in 13 cases of ESCC (13/18, n = 18) but absent in all of the normal tissues. The results indicated that there was a correlation between high level of UNG expression and the carcinogenesis of ESCC.
Carcinoma, Squamous Cell
;
genetics
;
metabolism
;
Cell Line, Tumor
;
Cloning, Molecular
;
Electrophoresis, Polyacrylamide Gel
;
Esophageal Neoplasms
;
genetics
;
metabolism
;
Humans
;
In Vitro Techniques
;
Reverse Transcriptase Polymerase Chain Reaction
;
Uracil-DNA Glycosidase
;
genetics
;
metabolism
6.Manipulative reduction with minimally invasive percutaneous plate osteosynthesis for 60 patients with distal tibiofibular fractures.
Bo-Chuan HAO ; Ke-Bo XIE ; Lin-Xiao XIONG ; Lei ZHANG ; Jun-Song ZHANG ; Jie ZHANG ; Yi-Bing ZHENG ; Li-Qiang ZHANG ; Shu-Ren BAO
China Journal of Orthopaedics and Traumatology 2014;27(6):491-495
OBJECTIVETo explore clinical effects of manipulative reduction with minimally invasive percutaneous plate osteosynthesis in treating distal tibiofibular fractures.
METHODSFrom 2009 to 2011, 60 patients with distal tibiofibular fractures were treated by manipulative reduction with minimally invasive percutaneous plate osteosynthesis. Among them, there were 32 males and 28 females aged from 14 to 70 years old with an average of 41.22 +/- 2.06. According to AO classification of fractures,5 cases were type A1, 22 cases were type A2,21 cases were type A3 and 12 cases were type C1. Operation time, blood loss,time of callus and fracture healing were observed, Mazur scoring of ankle joint were used to evaluate therapeutic.
RESULTSFifty-eight incisions were healed at stage I ,and 2 cases were infected at distal tibial. Operation time was with an average of (62.34 +/- 5.66) min ranged 45 to 90 min;blood loss was 30 to 150 ml with an average of (80.57 +/- 5.59) ml;formation of callus appeared from 4 to 12 weeks,with an average of (8.24 +/- 2.06) weeks, and fracture healing time was from 3 to 6 months, with an average of (4.50 +/- 1.13) months. According to Mazur scoring of ankle joint 40 cases got excellent results, 18 good, and 2 fair.
CONCLUSIONManipulative reduction with minimally invasive percutaneous plate osteosynthesis can obtain reliable fixation. It is a good choice of treating distal tibiofibular fractures by protecting blood supply of fractures.
Adolescent ; Adult ; Aged ; Bone Plates ; Female ; Fracture Fixation, Internal ; instrumentation ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Tibial Fractures ; surgery ; Treatment Outcome ; Young Adult
7.Multi-slices spiral CT perfusion imaging evaluating of microvessel density in maxillofacial tumors
Chuan-Ting LI ; Yu-Bo LV ; Dong-Sheng ZHANG ; Wei-Dong ZHANG ; Jing-Guang LV ; Cheng LIU ; Le-Bin WU ;
Chinese Journal of Radiology 2001;0(08):-
Objective To evaluate the relationship between Multi-slices spiral(MSCT) perfusion and microvessel density (MVD) in maxillofacial tumors. Methods Thirty-one cases of maxillofacial tumors were studied with MSCT perfusion imaging before operation. The time-density curve, perfusion, time to peak(TTP), and Peak enhancement imaging (PEI) of tumors were calculated. MVD of the tumors was measured with immuno-histochemical method by means of detecting factor Ⅷ in all the histologic specimens. Relativity analysis was carried between MSCT perfusion imaging parameters, perfusion curve types and MVD. Results MVD of maxillofacial tumors were higher than normal tissue. MVD remarkably correlated with malignancy of the tumors. Perfusion and time to peak (TTP) correlated well with MVD(t=7.09,4.10, P0.05). Significant difference of MVD in three types of perfusion curve was found(F=8.09,P
8.Type I collagen modified titanium sheet improves the proliferation of human adipose-derived mesenchymal stem cells
Chuan YE ; Minxian MA ; Tao ZHANG ; Jin TANG ; Bo SUN ; Long YANG ; Houxiang REN ; Qi SUN ; Jing YANG ; Junbiao ZHANG
Chinese Journal of Tissue Engineering Research 2014;(25):4032-4037
BACKGROUND:Titanium and titanium aloy are used mostly in artificial joints, fracture fixation, and oral transplantation, while there are complex cases of insufficient bone mass in these areas. The deepened research of stem cels offers a solution for bone injury to promote new bone formation. The biocompatibility of titanium and stem cels and optimization of titanium surface modification have aroused people's attention. OBJECTIVE:To investigate whether the biocompatibility of titanium and human adipose-derived mesenchymal stem cels can be improved by type I colagen modification of titanium sheets. METHODS:The experiment was divided into two groups. Modification group: titanium sheet was modified with type I colagen; control group: titanium sheet was not modified with type I colagen. Human adipose-derived mesenchymal stem cels at passage 6 were implanted into titanium sheet in two groups. Then we calculated the number of adherent cels in two groups at 1, 2 and 4 hours after implantation, and compared the celladhesion rate. MTT assay was used to observe the proliferation of cels on titanium sheet at 2, 4, 6 and 8 days after implantation. DNA and protein content of cels were detected at 3, 6, 9 days after implantation. The growth of human adipose-derived mesenchymal stem cels seeded upon the titanium sheets was observed under scanning electron microscope at 6 days. RESULTS AND CONCLUSION:When the cels were cultured for 1 hour and 2 hours, the number of adherent cels in the modification group was higher than in the control group (P < 0.05). The absorbance of cels in two groups was increased as the culture time, as detected by MTT assay. The modification group had a significantly higher absorbance value than the control group at 4, 6, 8 days (P < 0.05). DNA and protein contents of the cels in the modification group were higher than that in control group at 6 and 9 days (P < 0.05). At 6 days, the number of adherent cels and secretion of adherent stromal cellmatrix in the modification group were significantly better than that in control group, observed by scanning electron microscopy. Type I colagen modified titanium sheets have good surface activity and biocompatibility, and can promote the proliferation of human adipose-derived mesenchymal stem cels.
9.Research progress of the drug delivery system of antitumor platinum drugs with macrocyclic compounds.
Chuan-zhu GAO ; Yan ZHANG ; Ji CHEN ; Fan FEI ; Tian-shuai WANG ; Bo YANG ; Peng DONG ; Ying-jie ZHANG
Acta Pharmaceutica Sinica 2015;50(6):650-657
Platinum-based anticancer drugs have been becoming one of the most effective drugs for clinical treatment of malignant tumors for its unique mechanism of action and broad range of anticancer spectrum. But, there are still several problems such as side effects, drug resistance/cross resistance and no-specific targeting, becoming obstacles to restrict its expanding of clinical application. In recent years, supramolecular chemistry drug delivery systems have been gradually concerned for their favorable safety and low toxicity. Supramolecular macrocycles-platinum complexes increased the water solubility, stability and safety of traditional platinum drugs, and have become hot focus of developing novel platinum-based anticancer drugs because of its potential targeting of tumor tissues/organs. This article concentrates in the research progress of the new drug delivery system between platinum-based anticancer drugs with three generations of macrocycles: crown ether, cyclodextrin, cucurbituril and calixarene.
Antineoplastic Agents
;
pharmacology
;
Calixarenes
;
Crown Compounds
;
Cyclodextrins
;
Drug Delivery Systems
;
Humans
;
Macrocyclic Compounds
;
pharmacology
;
Neoplasms
;
drug therapy
;
Platinum Compounds
;
pharmacology
10.Treatment of lumbar disk herniation with MRI guidance injection of oxygen-ozone
Cheng-Li LI ; Le-Bin WU ; Ji-Qing SONG ; Yu-Bo LU ; Yu-Jun XU ; Chuan-Chen ZHANG ;
Chinese Journal of Radiology 1994;0(06):-
Objective To evaluate the applied value of percutaneous oxygen-ozone injection in the treatment of lumbar disc herniation under open 0.23 T MRI guidance.Methods Mounted with ipath 200 optical tracking system,MR-guided injection of oxygen-ozone were performed via a medial border of the articular processes approach in 73 patients with clinically diagnosed LDH.MR compatible 19.5G or 21.0 G biopsy needle was used. Discography was performed in order to select indication before injection oxygen- ozone into nucleus pulposus in 26 patients.Sixty-four patients were injected to three sites:(1)Six to 10 ml oxygen-ozone was injected into discs centers,injected and suctioned alternately in order to make nucleus pulposus oxidation thoroughly.(2)The needle was withdrawn according to the scale of biopsy needle and optical tracking.Then,10 ml oxygen-ozone was injected into disc herniation. (3)After that,needle was withdrawn further about 1.0—1.5 cm to outside of annulus fibrosus.Fifteen to 20 ml oxygen-ozone was injected into intervertebral foramina around nerve roots.The oxygen-ozone concentration was 35—45?g/ml. Nine patients were only performed injection of oxygen-ozone into around nerve root,while not injection oxygen-ozone to nucleus pulposus for considering bad curative effect after discography.Results All of 73 patients were successfully local targeted and treated under MRI guidance without serious complications, such as nerve root injury.After 3—6 months follow-up,total overall efficacy was 91.3% with the excellent in 28,good in 39,and poor in 6,respectively.Conclusion Open MR-guided injection of oxygen-ozone, mounted with optical tracking system,is a safe and effective minimally invasive therapy for treating LDH.