1.Expression of Recombinant sPDGFR?-Fc in CHO and Its Anti-proliferation Analysis
Yan WAN ; Li-Ling LI ; Qiu-Ling XIE ; Shu-Jun GUO ; Li QIN ; Yong-Cang ZHANG ; Xiao-Jia CHEN ;
China Biotechnology 2006;0(07):-
Orjective:To obtain recombinant CHO-K1 with expressing sPDGFR? and to identify the biological activities of sPDGFR? secreted in non-serum medium.Methods:Recombinant human sPDGFR? expression vector pIRES-Neo3-sPDGFR?-Fc was constructed and then transfected into CHO-K1 cells by using LipofectamineTM 2000.After screened with G418 in 8 weeks,some monoclone cells were selected randomly to amplify in 96-well-plate to 24-well-plates,and then to identify positive cell clones by RT-PCR.Furthermore,the candidate cell clones were test by Real-Time PCR and Western blot assays.Finally,anti-proliferation activities of the expressed sPDGFR? were analyzed by MTT.Results:sPDGFR?-Fc was cloned into pIRES-Neo3 correctly.The sPDGFR?-Fc expression level in recombinant CHO-K1 cell clones were concordant in between Realtime PCR and Western blot assay.sPDGFR?-Fc obtained from cultured non-serum medium of positive CHO-K1 could significantly inhibit proliferation of vascular endothelial cell.Conclusion:Successed to select recombinant CHO-K1 cell lines with high expressed sPDGFR?-Fc.The sPDGFR?-Fc can inhibit the cell proliferation significantly and it means sPDGFR?-Fc might be a new anti-cancer drug in the future.
2.Effect of hyperbaric oxygen on the changes of autophagy and apoptosis in cerebral cortex in rats with brain explosive injury
Hong TANG ; Jia-Chuan LIU ; Yong-Ming ZHANG ; Yan-Yan YANG ; Wen-Jiang SUN ; Liang-Chao ZHANG ; Bing-Cang LI
Chinese Journal of Neuromedicine 2010;09(10):1014-1017
Objective To observe the changes of autophagy and apoptosis in the cerebral cortex in rats with brain explosive injury and received hyperbaric oxygen (HBO) treatment, and investigate the effect and significance of HBO on them. Methods Fifty-four SD rats were randomly divided into control group (n=6), explosive injury group (n=24) and explosive injury treated with HBO group (n=24).The control group was not injured and 600 mg TNT electric detonators were exploded over the brain of rats at a 12 cm distance in the explosive injury group and explosive injury treated with HBO group. HBO management was given to the explosive injury treated with HBO group 3 and 22 h after explosive injury and at the same time of the other 6 d. RT-PCR and Western blotting were used to detect the changes of Beclin-1 and caspase-3 in the cerebral cortex on the 6th and 24th h, 3rd and 7th d of injury. Results The expression levels of Beclin-1 and caspase-3 in the explosive injury group and explosive injury treated with HBO group were obviously higher than those in the control group (P<0.05). The levels of Beclin-1and caspase-3 in the explosive injury treated with HBO group were significantly lower than those in the explosive injury group at the same time points (P<0.05). Conclusion The decreased expressions of autophagy and apoptosis in brain cells by HBO treatment might be one of the possible mechanisms of treating craniocerebral injury.
3.Application of WAIS-RC short forms and adult intelligence disability scale in mental impairment assessment.
Yan-Xia PANG ; Jian ZHANG ; Cheng-Long YANG ; Yong CANG ; Xue-Ling WANG
Journal of Forensic Medicine 2011;27(3):189-192
OBJECTIVE:
Study on the application of WAIS-RC short forms and adult intelligence disability scale in mental impairment assessment.
METHODS:
Mental impairment assessment cases between July 2009 and March 2011 in judicial appraisal institute of Taizhou University were collected. Assessment results obtained with the WAIS-RC short forms and adult intelligence disability scale were compared with the experts assessing conclusions and analyzed using SPSS 11.5 software.
RESULTS:
Assessment results with the two scales did not fully comply with the expert's conclusions, with reliability coefficient were 0.785 and 0.940 respectively, correlation coefficient were 0.850 and 0.922 respectively.
CONCLUSION
The intelligence assessment was influenced by many factors. When the appraised individuals had nerve dysfunction and mild intelligence disability or mental disorders, the two scales should be used together. When the appraised individuals had moderate intelligence disability or mental disorders, adult intelligence disability scale had advantage.
Accidents, Traffic
;
Adolescent
;
Adult
;
Aged
;
Aged, 80 and over
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Brain Injuries/complications*
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Disability Evaluation
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Expert Testimony
;
Female
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Forensic Psychiatry
;
Humans
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Intellectual Disability/psychology*
;
Intelligence
;
Male
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Mental Disorders/psychology*
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Middle Aged
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Reproducibility of Results
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Severity of Illness Index
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Wechsler Scales/statistics & numerical data*
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Young Adult
4.Human enterovirus 71 that firstly isolated in Qinghai Province and their genetic features of VP1 region.
Sheng-Cang ZHAO ; Shuang-Ying JIANG ; Gui-Xiang LIU ; Hu YI ; Yong ZHANG
Chinese Journal of Experimental and Clinical Virology 2010;24(6):436-438
OBJECTIVETo study the genetic characterizations of VP1 gene of human enterovirus 71 (HEV71) isolated from clinical specimens of Hand, Foot and Mouth Disease (HFMD) patients in Qinghai Province in 2008.
METHODS335 clinical samples including stools, throat swabs and vesicle fluids were collected from HFMD patients in Qinghai Province. Viral isolation was performed, and molecular typing was performed with the positive isolates. Then 30 identified HEV71 isolates were performed for entire VP1 coding region amplification and sequencing.
RESULTSAmong the 355 clinical samples, 45 human enteroviruses were isolated, and among them, 30 were identified as HEV71. Then 30 HEV71 positive isolates were performed by nucleotide sequencing. It showed that there was some difference in the nucleotide and the amino acid among the 30 HEV71 strains, the homology were 95.2%-100% and 96.6%-100%, respectively. But they all closed to HEV71 strains isolated in China after 1998, and from the phylogenetic tree constructed with 30 Qinghai HEV71 strains and other 35 HEV71 strains represented all known genotype and subgenotype HEV71 strains available from GenBank, it revealed that the 30 Qinghai HEV71 strains clustered within the C4a evolution branch of C4 subgenotype.
CONCLUSIONHEV71 was isolated in HFMD patients in Qinghai province, and the HEV71 strains causing HFMD outbreaks in Qinghai province in 2008 were all belong to C4a evolution branch of C4 subgenotype with several transmission chains.
China ; Enterovirus A, Human ; genetics ; isolation & purification ; Hand, Foot and Mouth Disease ; virology ; Humans ; Molecular Typing ; methods ; Sequence Analysis, DNA ; methods
5.Hyperbaric oxygen therapy in rabbits after explosive brain injury: a MRS and DWI study
Yan-Yan YANG ; Jia-Chuan LIU ; Yong-Ming ZHANG ; Wen-Jiang SUN ; Hong TANG ; Zhen-Shan HUANG ; Bing-Cang LI ; Liang-Chao ZHANG
Chinese Journal of Neuromedicine 2011;10(1):33-36
Objective To evaluate the role of magnetic resonance spectroscopy (MRS) and diffusion-weighted MRI (DWI) in detecting regional cerebral metabolic changes and changes of water molecular motion in rabbits after explosive brain injury at different time points of injury after being treated with hyperbaric oxygen therapy. Methods Ninety New Zealand white rabbits were divided into control group, trauma group and hyperbaric oxygen treatment group. The injured models in the later 2 groups were established using 600 mg TNT equivalent of paper detonators in the rabbit brain at the top of 6.5 cm vertical distance. Rabbits in the trauma group and hyperbaric oxygen treatment group were sub-divided into 1, 3, 7, 14 and 21 d treatment groups, respectively. The survival of these rabbits was observed at these time points. Hyperbaric oxygen treatments lasting for 1, 3, 7, 14 and 21 d were given to each hyperbaric oxygen treatment sub-groups, respectively. Performance under MRS was detected and dynamic changes of N-acetylaspartate (NAA)/creatine (Cr) ratio and NAA/choline(Cho)+Cr value were observed with MRS. DWI was employed to detect the dynamic changes of apparent diffusion coefficient (ADC) values. Results The NAA/Cr ratio in the trauma group markedly decreased right after the injury and slightly rose 7 d after the injury. Compared with that in the trauma group, the NAA/Cr ratio in the hyperbaric oxygen treatment group was significantly increased (P<0.05). The NAA/Cho+Cr value in the trauma group was markedly decreased right after the injury, while that in the hyperbaric oxygen treatment group was obviously increased as compared with that in the trauma group (P<0.05). ADC values in the region of interest of the trauma group was decreased after 1 d of treatment and gradually increased after 3 d of treatment; the ADC values in the hyperbaric oxygen treatment group was obviously higher than those in the trauma group (P<0.05). Conclusion Hyperbaric oxygen might improve the prognosis by improving local metabolism of neurons, inhibiting brain edema, and enhancing local gliosis repair;, ultra-early intervention (within one week of injury) may be much favorable for animals/patients after explosive brain injury.
6.Expression of estrogen receptor α in hepatitis B-related hepatocellular carcinoma and its clinical significance.
Sheng-jin HAN ; Ge-liang XU ; Wei-dong JIA ; Yong-cang WANG ; Jian-sheng LI ; Jin-liang MA ; Wei-hua REN ; Yong-sheng GE ; Wen-bin LIU ; Chuan-hai ZHANG ; Wei WANG
Chinese Journal of Surgery 2010;48(24):1875-1880
OBJECTIVETo investigate the expression and its clinical significance of estrogen receptor (ERα) and phosphorylated estrogen receptor (p-ERα) in patients with hepatocellular carcinoma. The associations between ERα, p-ERα and IL-6 were also analyzed.
METHODSImmunohistochemistry was used to detect the expression of ERα, p-ERα and IL-6 in tumor tissues from 77 cases with hepatocellular carcinoma. The relations between ERα and the clinical pathological parameters and prognosis were also analyzed.
RESULTSThe positive rates of ERα, p-ERα and IL-6 in hepatocellular carcinoma were 39.0% (30/77), 45.4% (35/77) and 72.7% (56/77), respectively. The expression of ERα and p-ERα were negatively correlated with the expression of IL-6 (r=-0.468, P<0.01; r=-0.370, P<0.01, respectively). The positive rate of ERα in patients with tumor size≤5 cm, serum level of alpha-fetoprotein<400 µg/L, with complete encapsulation and non-microvascular invasion was significantly higher than those with tumor size>5 cm, serum level of alpha-fetoprotein≥400 µg/L, non-complete encapsulation and with microvascular invasion (all P<0.05). The overall survival rates of ERα-positive and ERα-negative patients were 66.7% and 23.4% (P<0.05). And the disease-free survival rates of ERα-positive and ERα-negative patients were 83.3% and 57.4% (P<0.05).
CONCLUSIONSThe tumor biological features of ERα-positive patients are better than that of ERα-negative patients. The role of ERα in hepatocellular carcinoma may be related to IL-6 level.
Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Estrogen Receptor alpha ; metabolism ; Female ; Hepatitis B ; metabolism ; pathology ; Humans ; Interleukin-6 ; metabolism ; Kaplan-Meier Estimate ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Phosphorylation ; Prognosis ; Proportional Hazards Models ; Young Adult
7.The epidemiological characteristics and the source of infection of reemerge human rabies from 2012 to 2017 in Qinghai
Li-li XU ; Hua-yi ZHANG ; Hua-xiang RAO ; You-ju LEI ; Yong-hong LI ; Jin-hua ZHAO ; Hai-lan CAO ; Hua-xiang FENG ; Yan SHI ; Gui-xiang LIU ; Sheng-cang ZHAO
Chinese Journal of Disease Control & Prevention 2019;23(6):690-693,699
Objective To understand the epidemic situation and the source of infection of the reemerge human rabies in Qinghai. Methods We collected the data on human rabies and the data on the cases of multi- victims bitten by the identical dog, and also the laboratory data of the nucleoprotein ( N) gene of rabies virus from the samples which were detected by reverse transcription-polymerase chain reaction (RT-PCR) and direct immunofluorescence assay (DFA) from 2012 to 2017, to describe the epidemiological characteristics of human rabies and the prevalence of rabies virus in host animals, and to explore the source of infection of reemerge human rabies. Results A total of 7 human cases were reported in 2012-2017 in Qinghai province, among which 1 was bitted by wolf, 2 were bitted by stray dogs, 3 were bitted by domestic dogs which injured by stray dogs or wolfs. A total of 892 canine brain tissue samples were collected, from which 46 positive samples were detected with the positive rate of 5.16% (95% CI:3.70%-6.61%). The positive samples were collected from the nomadic region, which were consistent had the location of the human rabies. The samples collected from the cases of multi-victims bitten by the identical dog/animal had the positive rate of 73.08%, and 4 out of 7 human rabies were exposed to the cases of multi-victims bitten by the identical dog/animal. Genetic sequencing of the rabies virus detected from canine brain tissue samples were belong to China IV lineage, which was closely related to the Arctic clade. Conclusions The reemerging rabies happened in nomadic region of Qinghai province could be a consequence of spillover from wildlife especially from wolfs. The better surveillance system covering the human, livestock and wildlife should be set up to mitigate the rabies virus spread from the wildlife.
8.Genotype, environment and their interactions of functional components in Rheum palmatum.
Lei MENG ; Hui-Juan HU ; Tong SHANG ; Wen-Cang TIAN ; Ming-Ming ZHANG ; Yong-Jie LI ; Li LI ; Sheng-Li WEI
China Journal of Chinese Materia Medica 2018;43(12):2495-2502
Eight kinds of provenance of Rheum palmatum collected from 4 provinces Sichuan, Ningxia, Gansu, Shannxi as test materials, which were transplanted under 3 different environments by using complete randomized block design with three replicates. The contents of the chemical components was determined by HPLC. This study aimed at analyzing the effect of genotype, environment and their interactions on the 4 kinds of functional components (phenolic acids, bianthrone, free anthraquinones and combined anthraquinones) in 14 kinds of active components of R. palmatum, in order to provide a theoretical basis for the selection of cultivated R. palmatum in high quality producing area and excellent provenance. The functional components of R. palmatum were influenced by genotype and environment. The content of phenolic acids was mainly influenced by environment, and the other three kinds of functional components were affected by environment and their interactions. The proportion of environment was larger. The cultivation quality of R. palmatum should give priority to environment, then choose a provenance. Sichuan may be beneficial in accumulation of free anthraquinones in R. palmatum, Gansu may facilitate the binding of combined anthraquinone, phenolic acids and bianthrone content. Preliminary inference based on the content and proportion of efficacy components, P2 could be potential special medicinal germplasm that have function of heat-clearing and detoxifying drugs. P6 could be potential special medicinal germplasm that activate blood circulation to dissipate blood stasis. P7 and P1 could all be potential specialmedicinal germplasms that exist diarrhea attack characters. The results of this study have important guiding significance for the production of rhubarb precision medicinal materials.
9.Gene Mutation and Clinical Characteristics of Patients with Acute Leukemia.
Jie YANG ; Yong-Bin YANG ; Rui-Cang WANG ; Jing-Yue LIU ; Yan LI ; Jun YUAN ; Jie LI ; Xiao-Xia ZHANG ; Xiao-Han GAO ; Hong-Ling HAO
Journal of Experimental Hematology 2022;30(6):1693-1699
OBJECTIVE:
To investigate the characteristics of gene mutation, clinical characteristics and significance in acute leukemia (AL) patients.
METHODS:
The clinical data of 102 AL patients in Hebei General Hospital from September 2016 to September 2020 were collected and analyzed retrospectively, including the characteristics of gene mutation, age, peripheral blood cells, bone marrow blasts, leukemia subtypes and myeloperoxidase (MPO).
RESULTS:
The total gene mutation rate was 87.25% (89/102) in all 102 patients. A total of 275 gene mutations were detected, with an average of 2.70 gene mutations per patient. The most frequent mutations of 102 patients were as follows: CEBPA (6.91%), NPM1 and ASXL1(6.18%), TET2 (5.82%), DNMT3A (5.45%), IDH2 and FLT3-ITD (5.09%). Gene mutations often occurred simultaneously. CEBPA mutation occurred in 10 cases of M2 subtype, while TET2 mutation occurred in 9 cases of M2 subtype. Among the most common gene mutations in MPO low expression group, mutation rates of NPM1, DNMT3A, IDH2, SF related gene mutation and RUNX1 were significantly different than those in MPO high expression group (all P<0.05). Univariate analysis showed that age, NPM1, DNMT3A and FLT3-ITD had significant effects on leukocyte level. Logistic regression analysis showed that patients with positive NPM1 mutations may had higher leukocyte levels (p=0.038), and those with positive DNMT3A mutations may had higher platelet levels (p=0.042).
CONCLUSION
The incidence of gene mutation in patients with AL is high, and it often occurs simultaneously. CEBPA and TET2 gene mutations are more common in M2 subtype. In patients with MPO low expression, the most common gene mutations are NPM1, DNMT3A and IDH2. AL patients with NPM1 gene mutation had higher white blood cell levels, while with DNMT3A gene mutation had higher platelet levels.
Humans
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Retrospective Studies
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Leukemia
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Mutation
10.Study on geographical variation of medicinal ingredients contents in Scutellaria baicalensis from different provenances.
Yong-Jie LI ; Rong-Xiu LIU ; Wen-Cang TIAN ; Xue-Sen WANG ; Zhi-Yan HOU ; Lei MENG ; Guo-Hu SONG ; Xue-Wen ZHANG ; Sheng-Li WEI
China Journal of Chinese Materia Medica 2018;43(13):2664-2669
In order to investigate the genetic difference on medicinal components of Scutellaria baicalensis from different provenances on the genetic difference, the S. baicalensis provenance tests were arranged by randomized block design.Excavating the crude drugs that have been growing for three years, with the same drying process, the content of baicalin, baicalein, wogonoside, wogonin and laminarin A in S. baicalensis were detected by HPLC, and then the data were analyzed. The results indicated that the content of baicalin in different provenances of S. baicalensis was significantly different (<0.05), while the variation of baicalein reached extremely significant level (<0.01). Cluster analysis showed that if the distance was divided by 5.0, the provenances in Chengde, Hebei province were divided into two independent populations, while the other two populations had large geographic spans. The results show that the significant geographical variations exist in the content of medicinal components in S. baicalensis. The study laid a theoretical foundation of provenance selection of S. baicalensis.