2.Expression of collagen subtype I mRNA during autogenous tendon formation induced by grafted human hair keratin artificial tendon
Li LOU ; Weiren DONG ; Yingqing XIAO ; Qiaoyan CHEN ; Yingjie PIAO
Chinese Journal of Tissue Engineering Research 2005;9(22):213-215
BACKGROUND: Numerous experiments and clinical practice show that human hair keratin artificial tendon induces the organism to form autogenous tendon. The process of autogenous tendon formation mainly involves the synthesis, secretion and package of collagen subtype I.OBJECTIVE: To explore the role of collagen subtype I mRNA expression in autologous tendon formation after human hair keratin artificial tendon implantation.DESIGN: A completely randomized controlled experiment based on the experimental animals.SETTING: Department of Histology and Embryology and Department of Biochemistry and Molecular Biology of Southern Medical University.MATERIALS: The experiment was conducted in the Experimental Animal Center of the First Military Medical University of Chinese PLA from May 2003 to September 2004. Totally 33 New Zealand rabbits of either gender,weighing 2.0 to 2. 5 kg, were provided by the center. The animals were randomly divided into experiment 3, 6, 9, 12, 16 and 20 weeks groups, negative control 9 and 20 weeks groups and normal control group. Among them,experiment 3 and 6 weeks group and normal control group had 3 rabbits in each and the other groups had 4 rabbits. Human hair keratin artificial tendons were normal human hair treated by a series of biochemical methods and were supplied by the Department of Biochemistry & Molecular Biology of the university. The human hair keratin artificial tendons were divided into three groups with different degradation rates, namely, fast(F), medium(B) and slow(Z). The tendons were made up of the fast, medium and slow degradation groups mixed at the ratio of 4: 3: 3.off by 1.0- 2.0 cm, human hair keratin artificial tendon was grafted by end-to-end anastomosis with both ends of the broken tendon before sewing control group, no artificial tendon was implanted although the animals ungroup was normal rabbits' tendon. Sampling was carried out at 3, 6, 9, 12, 16and 20 weeks after human hair keratin artificial tendon implantation in experiment groups, and at 9 and 20 weeks after operation in negative control group, respectively. The expression of collagen subtype I mRNA was detected at weeks 3, 6, 9, 12, 16 and 20 after grafting using reverse transcription polymerase chain reaction technique.MAIN OUTCOME MEASURES: The ratio of collagen subtype I mRNA to Glyceraldehyde-3-phosphate dehydrogenase(GADPH) mRNA in normal tendon and autogenous tendon induced by human hair keratin artificial tendon at all time points was calculated, and significance test between all these paired groups were performed.collagen subtype I mRNA/GADPH mRNA expression was 0.96 ±0.02 in expression of collagen subtype I mRNA/GADPH mRNA in autogenous tendon induced by human hair keratin artificial tendon in experiment group appeared at week 3, increased rapidly at week 3 to 6, peaked at week 6, and remained stable at week 9 to 20. The expression at week 6 was significantly higher in experiment group than in normal control group( F = 6. 254, P < 0.05); the expression at other weeks was also significantly higher in experiment group than in normal control group( F= 1. 258 - 1. 987, P > 0.05).CONCLUSION: The activation, proliferation and secretion of collagen protein as well as the synthesis of collagen subtype I by tenocytes may be responsible for autologous tendon formation after human hair keratin artificial tendon implantation.
3.Influence of Montelukast on Plasma Nitric Oxide in Preschool Children with Asthma
huan-yin, YAO ; xiao-qin, YAN ; wen-yong, LOU ; shu-mei, LIU ; chuan-chang, DONG
Journal of Applied Clinical Pediatrics 2006;0(22):-
Objective To explore the influence of montelukast on plasma nitric oxide in preschool children with asthma.Methods Forty-four preschool children with asthma aged 2-5 years who firstly met a criterion of asthma and treated 4 weeks with montelukast were investigated;and nitric oxide levels of plasma were inspected respectively before treatment and after treatment 1 week,4 weeks.Results The level of nitric oxide in the plasma of asthmatic children was obviously higher than that in normal control group(P
4.Keratinase production and keratin degradation by a mutant strain of Bacillus subtilis.
Cheng-gang CAI ; Bing-gan LOU ; Xiao-dong ZHENG
Journal of Zhejiang University. Science. B 2008;9(1):60-67
A new feather-degrading bacterium was isolated from a local feather waste site and identified as Bacillus subtilis based on morphological, physiochemical, and phylogenetic characteristics. Screening for mutants with elevated keratinolytic activity using N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis resulted in a mutant strain KD-N2 producing keratinolytic activity about 2.5 times that of the wild-type strain. The mutant strain produced inducible keratinase in different substrates of feathers, hair, wool and silk under submerged cultivation. Scanning electron microscopy studies showed the degradation of feathers, hair and silk by the keratinase. The optimal conditions for keratinase production include initial pH of 7.5, inoculum size of 2% (v/v), age of inoculum of 16 h, and cultivation at 23 degrees C. The maximum keratinolytic activity of KD-N2 was achieved after 30 h. Essential amino acids like threonine, valine, methionine as well as ammonia were produced when feathers were used as substrates. Strain KD-N2, therefore, shows great promise of finding potential applications in keratin hydrolysis and keratinase production.
Amino Acids
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biosynthesis
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Bacillus subtilis
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genetics
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metabolism
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Culture Media
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Hydrogen-Ion Concentration
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Keratins
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metabolism
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Mutation
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Peptide Hydrolases
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biosynthesis
5.The epidemic situation of human rabies from 1984 to 2002 and its preventive measures in China.
Yong-Zhen ZHANG ; Dong-Lou XIAO ; Yu-Hui SUN ; Xiao-Ran YANG ; Yan-Zhen YAN
Chinese Journal of Epidemiology 2003;24(10):883-888
OBJECTIVETo summarize and analyze the epidemic situation of human rabies from 1984 to 2002 in China, and to explore the possible factors causing the increase of cases so as to provide evidence for preventive and control measures.
METHODNational and some provincial data on the prevalence of rabies during 1984 to 2002 were collected and analyzed.
RESULTSFrom 1984 to 1989, the annual reported cases were between 4 000 and 6 000 but decreased after 1990. In 1996, the reported cases decreased to the lowest level from 3 520 in 1990 to 159. However, number of reported cases has been continuously increasing since 1998 which reached 1 122 in 2002, a 7.06 times increase as compared to the number in 1996. The epidemic areas were mainly located in the southeast and southwest parts of the country, such as Sichuan, Hunan, Guangxi, Guangdong, Anhui, Fujian, etc. Furthermore, there was no significant seasonal distribution as it showed before.
CONCLUSIONSuch facts as the increasing numbers of dogs, low inoculation rate to dogs, poor control on the quality of rabies vaccine, mistreatment to the wounds, and lacking good cooperation between different official departments regarding rabies control might serve as important factors responsible for the recurrence of rabies. Therefore, it is necessary to focus on the above mentioned points and to take comprehensive preventive measures to bring down the prevalence of rabies in China.
Animals ; China ; epidemiology ; Dogs ; Humans ; Rabies ; epidemiology ; prevention & control ; Rabies Vaccines ; standards ; Seasons ; Time Factors
6.Promotive effect of LRP16 gene on proliferation of K562 cells.
Bo YANG ; Xue-Chun LU ; Xiao-Hua CHI ; Wei-Dong HAN ; Li YU ; Fang-Ding LOU
Journal of Experimental Hematology 2009;17(5):1154-1158
The study was aimed to investigate the promotive effect of LRP16 gene on K562 cell proliferation. Open reading frame of LRP16 gene was amplified using reverse transcription-polymerase chain reaction (RT-PCR) and ligated to pGEM-T plasmid to construct LRP16 ORF-pGEM-T recombinant vector. Then, LRP16 ORF identified by sequencing was inserted into pcDNA3.1+ plasmid to construct LRP16 ORF-pcDNA3.1+ recombinant expression plasmid which was transfected into K562 cell lines to make overexpression of LRP16 gene in K562 cells. Survival of cells was determined by MTT assay and growth curve of cells was drawn, the cell cycle was detected by flow cytometry. The results showed that LRP16 ORF was successfully amplified, then the LRP16 ORF-pcDNA3.1+ recombinant plasmid was constructed. The K562 cell line with overexpression of LRP16 gene was established. The promotive effect of LRP16 gene overexpression on proliferation of K562 cells was observed and the effect partially related to the enhancement of cells from G0 to S phase induced by LRP16 gene. It is concluded that LRP16 gene overexpression shows a promotive effect on proliferation of K562 cells.
Cell Proliferation
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Genetic Vectors
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Humans
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K562 Cells
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Neoplasm Proteins
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genetics
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Open Reading Frames
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Plasmids
7.A preliminary report on treatment of myelodysplastic syndrome with cyclosporin a and androgen.
Jing-Wen WANG ; Feng NING ; Xiao-Dong LIU ; Jin-Xing LOU ; Peng CHEN ; Bing LIU
Journal of Experimental Hematology 2004;12(6):855-857
The study was aimed to explore clinical result of cyclosporin A (CsA) and androgens for treatment of myelodysplastic syndrome (MDS) with refractory anemia. Four cases of MDS-RA were treated with CsA and androgens, while the changes of blood counts, bone marrow and chromosome were observed. The results showed that substantial hematological response was observed in all four patients, that their anemia improved and all transfusion-dependent patients achieved transfusion independence. In conclusion, CsA and Adr therapy was well tolerated. CsA and Adr therapy offer an alternative treatment of MDS with RA. The mechanisms of the benifical effect from this therapy remain the subject of an ongoing study.
Adolescent
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Adult
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Aged
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Androgens
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therapeutic use
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Blood Cell Count
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Cyclosporine
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therapeutic use
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Drug Therapy, Combination
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Female
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Hemoglobins
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analysis
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Humans
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Immunosuppressive Agents
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therapeutic use
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Male
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Middle Aged
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Myelodysplastic Syndromes
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blood
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drug therapy
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Treatment Outcome
8.Lrp16 gene expression in leukemia cell lines and bone marrow cells of leukemia patients and its clinical implication.
Bo YANG ; Xiao-Hua CHI ; Xue-Chun LU ; Wei-Dong HAN ; Li YU ; Fang-Ding LOU
Journal of Experimental Hematology 2009;17(4):857-860
This study was purposed to investigate lrp16 gene expression in leukemia cell lines and bone marrow cells of leukemia patients and explore the relationship between lrp16 gene expression and development of leukemia. Reverse transcriptase-polymerase chain reaction (RT-PCR) was employed to test the lrp16 mRNA expression in 4 leukemia cell lines, including K562 (CML), HL-60 (APL), MOLT4 (ALL) and U937 cell lines, as well as in bone marrow-derived cells from 115 patients with leukemia. The effect of lrp16 gene expression on genesis and progression of leukemia was analyzed according to clinicopathological features. The results indicated that positive expression of lrp16 mRNA was found in all 4 leukemia cell lines. For leukemia patients, the positive expression rate of lrp16 mRNA in all AML patients was 38% (16/42), in which the positive rates in AML patients with complete remission (CR) and AML patients without remission were 13% (4/30) and 100% (12/12) respectively. The positive expression rate of lrp16 mRNA in ALL patients was 38% (10/26), in which the positive rate in ALL patients with CR and ALL patients without remission were 16% (3/18) and 87% (7/8) respectively. The positive expression rate of lrp16 mRNA in CML patients was 36% (9/25), in which the positive rates in CML patients with CR and CML patients without remission were 20% (4/20) and 100% (5/5) respectively. The positive rate of lrp16 mRNA in CLL patients was 31% (7/22), in which the positive rate in CLL patients with CR and CLL patients without remission were 11% (2/17) and 100% (5/5) respectively. There was no difference of lrp16 gene expression between leukemia subtypes, but there was statistical significant difference in lrp16 gene expression between CR patients and non CR patients (p < 0.001). It is concluded that the lrp16 gene is a leukemic oncogene and closely relates to genesis and progression of leukemia, which may be an indicator for evaluating clinical efficacy of leukemia therapy.
Adolescent
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Adult
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Aged
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Bone Marrow
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metabolism
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pathology
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Female
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HL-60 Cells
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Humans
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K562 Cells
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Leukemia
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metabolism
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pathology
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Male
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Middle Aged
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Neoplasm Proteins
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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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Young Adult
9.Study on mechanisms of the expression regulation of interferon-induced gene RIG-G.
Dong LI ; Shu XIAO ; Xiao-rong PAN ; Ye-jiang LOU ; Pei-min JIA ; Jian-hua TONG
Chinese Journal of Medical Genetics 2007;24(6):625-628
OBJECTIVETo investigate the molecular mechanisms of the expression regulation of retinoic acidinduced gene G (RIG-G) by interferon alpha (IFNalpha).
METHODSRIG-G promoter region was analyzed by bioinformatics. The functional activities of RIG-G promoter with or without IFNalpha were detected by luciferase reporter assay and electrophoretic mobility shift assay (EMSA).
RESULTSRIG-G promoter region contained two well-conserved IFN-stimulated response elements (ISREs). Both ISRE I and ISRE II showed their effective binding abilities with signal transducer and activator of transcription 1 (STAT1). In HT1080 cells, in contrast with the empty plasmid pXP2, pXP2-A reporter construct containing intact ISRE I and ISRE II showed a significant higher baseline expression (1741.2 +/- 517.5) which could be further enhanced up to three-four folds by IFNalpha (5338.7 +/- 1226.9, P < 0.05). However, the luciferase activity of pXP2-A as well as its IFNalpha inducibility could be abrogated in STAT1-deficient U3A cells (from 1741.2 +/- 517.5 to 406.1 +/- 103.2, P < 0.05), indicating that the STAT1 protein was a prerequisite for the activities of ISRE I and ISRE II.
CONCLUSIONISREs present in RIG-G promoter region are molecular basis of IFNalpha induced RIG-G expression. RIG-G is a target gene directly regulated by STAT1 protein and should play a key role in IFNalpha signaling pathways.
Base Sequence ; Cells, Cultured ; Gene Expression Regulation ; drug effects ; physiology ; Humans ; Interferon Regulatory Factor-1 ; genetics ; metabolism ; Interferon Regulatory Factors ; genetics ; metabolism ; Interferon-alpha ; pharmacology ; physiology ; Interferons ; physiology ; Molecular Sequence Data ; Promoter Regions, Genetic ; drug effects ; genetics ; physiology ; STAT1 Transcription Factor ; metabolism
10.The epidemic characteristics and preventive measures of hemorrhagic fever with syndromes in China.
Yong-Zhen ZHANG ; Dong-Lou XIAO ; Yu WANG ; Hong-Xia WANG ; Li SUN ; Xiao-Xia TAO ; Yong-Gang QU
Chinese Journal of Epidemiology 2004;25(6):466-469
OBJECTIVETo summarize and analyze the epidemic situation of hantaviruses including geographic distribution, types and prevalent intensity of epidemic areas of hantavirus for the last 30 years in China, and to discuss relative preventive measures.
METHODSCollecting and analyzing the data of hantaviruses epidemics in China.
RESULTSThe annual number of cases of hantavirus disease rapidly increased from 3295 in 1970 to 115,804 in 1986 then sustained between 40,000 and 60,000 cases annually in the 1990's, and then decreased thereafter. The epidemic areas existed in all provinces except Qinhai and Xinjiang and there were the hospitalized cases of hantavirus disease reported in other provinces. In recent years, the prevalence of hantavirus infection had increased in some cities, and the seasonal distribution of the cases changed as well.
CONCLUSIONData suggested that the new epidemic characteristics of hantaviruses had emerged in China suggesting that it was necessary to strengthen surveillance programs and to take comprehensive preventive measures for the control and prevention of hantaviruses in China.
Animals ; China ; epidemiology ; Disease Reservoirs ; Female ; Hantaan virus ; immunology ; Hemorrhagic Fever with Renal Syndrome ; epidemiology ; prevention & control ; transmission ; Humans ; Male ; Mice ; Population Surveillance ; Prevalence ; Rats ; Rodent Control ; Vaccination ; Vaccines, Inactivated ; immunology