1.The influence of STK31 on osteosarcoma's malignant biological behavior
Jian MO ; Jianglong ZHU ; Zhe FENG ; Shuzhen LI ; Jie HAN ; Bo SU ; Sitan FENG
Chongqing Medicine 2017;46(23):3195-3197,3200
Objective To observe the expression of serine/threonine kinase 31 (STK31) in osteosarcoma and its effect on the malignant biological behavior of osteosarcoma.Methods Fifteen cases of osteosarcoma specimens and adjacent normal tissue were collected.The expression of STK31 in tumor tissues and normal tissue were detected by immunohistochemistry,real-time quantitative PCR and Western blot.The STK31 knockout plasmids PGenesil-STK31-shRNA or control plasmid pGenesil-1 were transfected into osteosarcoma cell line MG63 cells.The effect of STK31 on the proliferation of MG63 cells was detected by CCK8 cell activity assay.Tanswell experiment was used to observed the effect of STK31 on the migration ability of osteosarcoma cells.Results Immunohistochemical showed that STK31 expressed in the tumor tissue,and it was significantly higher than the adjacent normal tissues;Real time quantitative PCR[(3.65±0.83)vs.(1.05±0.14),P<0.05] and Western blot also revealed that STK31 expression in tumor tissue were significantly higher than adjacent normal tissues(P<0.05);CCK8 experiments showed that knockdown STK31 inhibited proliferation of MG63 cell when compared with the control group after 36 h[(1.71±0.17)vs.(1.39±0.11),P<0.05],72 h[(2.15±0.21)vs.(1.54±0.14),P<0.05];Tansewell experiments showed that transfection of pGenesil-STK31-shRNA could suppress MG63 cell's migration[(13±4)vs.(55±8),P<0.05].Conclusion STK31 is overexpression in osteosarcoma with increased biological activity of osteosarcoma cells.
2.Application of mesenchymal stem cells as a vehicle to deliver replication-competent adenovirus for treating malignant glioma.
Cui HAI ; Yong-Min JIN ; Wen-Biao JIN ; Zhe-Zhu HAN ; Mei-Nv CUI ; Xue-Zhe PIAO ; Xiong-Hu SHEN ; Song-Nan ZHANG ; Hong-Hua SUN
Chinese Journal of Cancer 2012;31(5):233-240
Although gene therapy was regarded as a promising approach for glioma treatment, its therapeutic efficacy was often disappointing because of the lack of efficient drug delivery systems. Mesenchymal stem cells(MSCs) have been reported to have a tropism for brain tumors and thus could be used as delivery vehicles for glioma therapy. Therefore, in this study, we attempted to treat glioma by using MSCs as a vehicle for delivering replication-competent adenovirus. We firstly compared the infectivity of type 3, type 5, and type 35 fiber-modified adenoviruses in MSCs. We also determined suitable adenovirus titer in vitro and then used this titer to analyze the ability of MSCs to deliver replication-competent adenovirus into glioma in vivo. Our results indicated that type 35 fiber-modified adenovirus showed higher infectivity than did naked type 3 or type 5 fiber-modified adenovirus. MSCs carrying replication-competent adenovirus significantly inhibited tumor growth in vivo compared with other control groups. In conclusion, MSCs are an effective vehicle that can successfully transport replication-competent adenovirus into glioma, making it a potential therapeutic strategy for treating malignant glioma.
Adenoviridae
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Animals
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Brain Neoplasms
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pathology
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therapy
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Cell Line, Tumor
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Genetic Vectors
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Glioma
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pathology
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therapy
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Humans
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Mesenchymal Stem Cell Transplantation
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Mesenchymal Stromal Cells
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Mice
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Mice, Inbred BALB C
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Mice, Nude
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Neoplasm Transplantation
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Oncolytic Virotherapy
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Random Allocation
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Virus Replication
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Xenograft Model Antitumor Assays
3.Sequence analysis of a novel HLA-DRB1 allele, DRB1 * 1212.
Fa-ming ZHU ; Wei ZHANG ; Qin-feng LU ; Ji HE ; Wei WANG ; Zhe-dong HAN ; Li-xing YAN
Chinese Journal of Medical Genetics 2006;23(1):47-49
OBJECTIVETo investigate the molecular genetics basis for HLA novel allele HLA-DRB1*1212 in Chinese population.
METHODSGenomic DNA was extracted from whole blood by salting-out method. HLA-DRB1 gene exon 2 was amplified by PCR with group-specific primers from genomic DNA. PCR products were cut back from agarose gels and purified to sequence directly. The polymerase chain reaction-sequence specific oligonucleotide probes (PCR-SSO) was performed to confirm the mutations which were detected by sequencing in this study.
RESULTSThe sequencing results showed HLA-DRB1 alleles of the proband as DRB1*090102 and the novel allele. The sequences of the novel allele have been submitted to GenBank (AY899825). Through BLAST analysis, the novel allele was found to be different from DRB1*120101 at position 199A-->C in exon 2, that results in an amino acid change from Ile to Leu at codon 67.
CONCLUSIONThis allele is a novel and has been officially named as DRB1*1212 by the WHO Nomenclature Committee.
Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; ethnology ; DNA ; analysis ; Ethnic Groups ; Female ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Analysis
4.Experimental study on ectopic prefabrication of vascularized mandible graft with autogenous ribs.
Xiang WANG ; Zhe SHAO ; Han-zhong ZHANG ; Fei ZHU ; Hui SHEN ; Zheng-jun SHANG
Chinese Journal of Stomatology 2012;47(9):544-546
OBJECTIVETo explore a method for ectopic prefabrication of mandible with vascular pedicle.
METHODSCancellous bone blocks harvested from the dog ribs were packaged with mandible-shaped titanium mesh scaffold and implanted into latissimus dorsi of dog with thoracodorsal artery and vein through the scaffold. After 12 weeks, bone formation and vascularization were evaluated by gross inspection, histological examination and immunohistochemistry.
RESULTSVascularized mandible with thoracodorsal artery and vein were formed and histological staining and immunohistochemisty confirmed new bone formation and vascularization.
CONCLUSIONSThere is feasibility for ectopic prefabrication of vascularized mandible graft using cancellous ribs, which provides a new method for mandibular defect reconstruction. Experimental study on ectopic prefabrication of vascularized mandible graft with autogenous ribs.
Animals ; Bone Transplantation ; methods ; Dogs ; Female ; Male ; Mandible ; surgery ; Osteogenesis ; Reconstructive Surgical Procedures ; methods ; Ribs ; surgery ; Tissue Engineering ; methods ; Transplantation, Autologous
5.Sequence analysis of HLA-B*4061 allele newly found.
Wei ZHANG ; Qin-Feng LÜ ; Wei WANG ; Zhe-Dong HAN ; Fa-Ming ZHU ; Li-Xing YAN
Journal of Experimental Hematology 2006;14(6):1188-1190
The aim of this study was aimed to investigate the molecular genetic basis for a novel HLA allele, HLA-B*4061, in Chinese population. DNA was extracted from whole blood by salting-out method. The HLA-B exons 1 - 8 of the proband was amplified and the amplified product was cloned using TOPO TA cloning sequencing kit to split the two alleles apart. Both strands of exons 2, 3 and 4 of chosen colonies were sequencing. The PCR-SSP was performed to confirm the mutations detected by sequencing. The sequencing results showed HLA-B alleles of the proband as B*4601 and the novel allele. The sequences of the novel allele have been submitted to GenBank (DQ089628, DQ089629, DQ089630). After HLA blast analysis, the novel allele showed a single nucleotide mismatch with B*400101 in exon 2 at position 272 C-->A, as the results, changing amino acid from Ser to Tyr at codon 67. It is concluded that this allele is a novel one and has been officially named B*4061 by the WHO Nomenclature Committee.
Alleles
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Amino Acid Substitution
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Asian Continental Ancestry Group
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genetics
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Base Sequence
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China
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HLA-B Antigens
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genetics
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immunology
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Histocompatibility Testing
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Humans
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Molecular Sequence Data
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Sequence Analysis, DNA
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Sequence Homology, Nucleic Acid
6.Sanguinarine Attenuates Lipopolysaccharide-induced Inflammation and Apoptosis by Inhibiting the TLR4/NF-κB Pathway in H9c2 Cardiomyocytes
Yan-Yan MENG ; Yuan LIU ; Zhe-Fu HU ; Yao ZHANG ; Jian NI ; Zhen-Guo MA ; Hai-Han LIAO ; Qing-Qing WU ; Qi-Zhu TANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2018;38(2):204-211
The inflammatory response is involved in the pathogenesis of the most common types of heart disease.Sanguinarine (SAN) has various pharmacological properties such as anti-inflammatory,antioxidant,antibacterial,antitumor,and immune-enhancing properties.However,few studies have investigated the effects of SAN on lipopolysaccharide (LPS)-induced inflammatory and apoptotic responses in H9c2 cardiomyocytes.Therefore,in this study,H9c2 cells were co-treated with SAN and LPS,and the mRNA levels of pro-inflammation markers and the apoptosis rate were measured to clarify the effect of SAN on cardiac inflammation.The underlying mechanism was further investigated by detecting the activation of Toll-like receptor (TLR)4/nuclear factor-κB (NF-κB) signaling pathways.As a result,increased mRNA expression of interleukin (IL)-1 β,IL-6,and TNFα induced by LPS was attenuated after SAN treatment;LPS-induced apoptosis of H9c2 cardiomyocytes and cleaved-caspase 8,9,3 were all significantly reduced by SAN.Further experiments showed that the beneficial effect of SAN on blocking the inflammation and apoptosis of H9c2 cardiomyocytes induced by LPS was associated with suppression of the TLR4/NF-κB signaling pathway.It was suggested that SAN suppressed the LPS-induced inflammation and apoptosis of H9c2 cardiomyocytes,which may be mediated by inhibition of the TLR4/NF-κB signaling pathway.Thus,SAN may be a feasible therapy to treat sepsis patients with cardiac dysfunction.
7.Observation on therapeutic alliance with UDCA and glucocorticoids in AIH-PBC overlap syndrome.
Jiang-yi ZHU ; Yong-quan SHI ; Zhe-yi HAN ; Gui JIA ; Zeng-shan LI ; Xiao-feng HUANG ; Jian-hong WANG ; Rui-an WANG ; Xin-min ZHOU ; Ying HAN
Chinese Journal of Hepatology 2011;19(5):334-339
OBJECTIVETo observe the efficacy of ursodeoxycholic acid (UDCA) combined with glucocorticoids in the treatment of autoimmune hepatitis-primary biliary cirrhosis (AIH-PBC) overlap syndrome.
METHODS19 patients with AIH-PBC overlap syndrome were divided randomly into two groups: initiate combined group and initiate UDCA-monotherapy group. Biochemical responses and pathological features before and after treatment were analyzed retrospectively with student's t test, Wilcoxon rank sum test and Fisher's exact method.
RESULTSIn the initiate combination group, biochemical responses in terms of AIH features (ALT decline to normal, IgG is less than or equal to 16 g/L) and PBC features (ALP decline ≥ 40% or to normal) were achieved. In UDCA-monotherapy group, no statistical difference existed in biochemical responses before adding glucocorticoids, whereas the levels of ALT, AST, GLB and IgG decreased significantly when combined with glucocorticoids. No statistical difference of rates of biochemical responses eixted between the two groups, whereas variance could be seen in different pathological stages. Alleviation of inflammatory infiltration after therapy appeared in 3 patients.
CONCLUSIONCombination therapy of UDCA with glucocorticoids could be suitable for AIH-PBC overlap syndrome. Early treatment is of benefit for achieving better biochemical response and pathological improvement.
Adult ; Alanine Transaminase ; analysis ; Drug Therapy, Combination ; Female ; Glucocorticoids ; administration & dosage ; therapeutic use ; Hepatitis, Autoimmune ; complications ; drug therapy ; Humans ; Immunoglobulin G ; analysis ; Liver Cirrhosis, Biliary ; complications ; drug therapy ; Male ; Middle Aged ; Treatment Outcome ; Ursodeoxycholic Acid ; administration & dosage ; therapeutic use
8.Comparison between two diagnostic criteria for PBC-AIH overlap syndrome.
Jiang-Yi ZHU ; Xin-Min ZHOU ; Jian-Hong WANG ; Rui-An WANG ; Xiao-Feng HUANG ; Zhe-Yi HAN ; Yu CHEN ; Ying HAN
Chinese Journal of Hepatology 2011;19(2):132-133
Not Abstract.
9.Establish predictive model of colorectal cancer by using surface enhanced laser desorption/ionization-time of flight-mass spectrometry.
Yan-Han LAI ; Jian-Min XU ; Xin-Zhe YU ; Yun-Shi ZHONG ; Ye WEI ; Li REN ; De-Xiang ZHU ; Yin-Kun LIU ; Wei-Xin NIU ; Xin-Yu QIN
Chinese Journal of Surgery 2008;46(13):995-997
OBJECTIVETo establish serum proteome fingerprinting predictive models and search for proteins associated with colorectal cancer.
METHODSThirty-six randomly selected colorectal cancer patients and 36 cases with hernia or gall bladder diseases scheduled for elective operation were enrolled as cancer group and control group respectively. Peripheral venous blood samples were collected before the operations. Special serum protein or peptide fingerprint was investigated by using surface enhanced laser desorption/ ionization-time of flight-mass spectrometry (SELDI-TOF-MS) measurement after blood sample had been treated with weak cation exchange protein chip (CM10) for each case. The obtained data were analyzed by Biomarker Wizard software to screen serum proteome tumor markers and set up diagnosis predictive model for colorectal cancer. Blind validation of the model with 44 healthy controls and 88 colorectal cancer patients were carried out by using Biomarker Patterns Software.
RESULTSIn comparing colorectal cancer group with control group, 5 specific protein peaks (P < 0.05) were found. The predictive model had a sensitivity of 100% and a specificity of 97.2%. A sensitivity of 71.6% and a specificity of 72.7% was got with the blind validation. The specific protein peaks with a mass-to-charge ratio (m/z) of 8908 and 13,707 showed in all the results and it showed their strong relationship with colorectal cancer.
CONCLUSIONSThe predictive models built by the differences of serum proteome fingerprint could be a very useful diagnostic tool in colorectal cancer. Proteins with m/z of 8908 and 13,707 would possibly be the tumor markers of colorectal cancer.
Biomarkers, Tumor ; blood ; Blood Proteins ; analysis ; Colorectal Neoplasms ; blood ; diagnosis ; Female ; Humans ; Male ; Middle Aged ; Peptide Mapping ; Proteomics ; methods ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
10.Identification and sequence analysis of a null HLA-B allele HLA-B*5408N newly detected.
Qin-Feng LÜ ; Fa-Ming ZHU ; Wei ZHANG ; Jun-Jun HE ; Wei WANG ; Zhe-Dong HAN ; Li-Xing YAN
Journal of Experimental Hematology 2007;15(4):870-872
The study was purposed to investigate the molecular genetic basis for HLA novel allele HLA-B*5408N in Chinese population. DNA was extracted from whole blood by commercial DNA extraction kit, the HLA-B exons 2 - 4 of the proband was amplified by allele specific primers PCR and the amplified product was sequenced for exons 2, 3 and 4 bidirectionally. The sequencing results showed HLA-B alleles of the proband as B*1527 and the novel allele. The sequences of the novel allele have been submitted to Genbank (DQ295998, DQ295999, DQ296000). After blast analysis, the novel allele showed a single nucleotide mismatch with HLA-B*5401 in exon 3 at position 553 G-->T, which resulted in an amino acid changing from Glu to premature stop codon at position 161. No the HLA-B54 antigen specificity expression in the proband cells was found using HLA-AB serological Typing Trays. It is concluded that this allele is a novel null allele and has been officially named B*5408N by the WHO Nomenclature Committee.
Alleles
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China
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Exons
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genetics
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HLA-B Antigens
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genetics
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Humans
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Polymerase Chain Reaction
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Sequence Analysis, DNA