3.Cloning, expression and protein structure prediction of a novel SCF-TPO fusion gene
Nan LIU ; Yongzhi XI ; Siqi GUO ; Yuying SUN ; Zhihong YUAN ; Jianwu CUI ; Caixia XI ; Fei LIANG ; Fanhua KONG
Chinese Journal of Pathophysiology 2000;0(10):-
AIM: To obtain the high expression of recombinant human stem cell factor - thrombopoitin (SCF-TPO) fusion gene and predict its structure property. METHODS: Tour primers were designed according to known sequence of TPO and SCF to amplify the functional amino acid domain of TPO and SCF by RT- PCR, respectively from fetus hepatocytes. The expression plasmid pET32a/SCF- TPO was constructed by VOE gene fusion technique and expressed in BL21(DE3)plysS. The fusion protein property, such as second structure, flexibility, and hydrophilicity were predicted by DS Gene and Protscale software. RESULTS: The expression vector, pET32a/SCF - TPO was constructed and the high expression of the SCF/TPO fusion protein was obtained, with the expression amount of up to 40% of the total cellular protein. DS Genel .5 and Protscale predict no new antigenicity in fusion protein, and the second structure and ioelectric point have no changes except four amino acids change in first structure. There are high flexibility and low hydrophilieity in the linker peptide. CONCLUSION: High expression of SCF- TPO fusion protein has been obtained and protein prediction shows that the fusion protein design is reasonable, which lay foundation for further study of biological fundation of SCF - TPO fusion protein.
4.Effect of soybean protein in lowering human serum cholesterol
Shiguang LI ; Kui FU ; Chunru QI ; Xianguo WU ; Qinghao KONG ; Weiping XI ; Yingyi LIANG
Chinese Journal of Tissue Engineering Research 2005;9(43):170-171
BACKGROUND: Although we have gained much information about leadinduced organ damage, the effect of blood lead level on T cell subgroup is yet to be determined.OBJECTIVE: To assess the effect of blood lead level on T cell subgroup of children, and the association of T cell subgroup with threshold limit value of blood lead level.DESIGN: Comparative observation.SETTING: The Institute of Molecular Microorganism, Maternity and Child Care Hospital, Shanxi Provincial Children Hospital.PARTICIPANTS: Sixty children, (32 boys and 28 girls), aged 3-6 years with a mean of (4.5±1.5) years old, were selected from the suburbs of Taiyuan city between May 2003 and October 2003. Informed consents were obtained from all their guardians. The enrolled children according to their blood lead levels were assigned into three groups, 13 in Group Ⅰ with blood lead level ≥ 0.48 μmol/L, 20 in Group 2 with lead level ≥ 0.24 μmol/L but < 0.48 μmoL/L and 27 in Group 3 with lead level < 0.24 μmol/L.METHODS: Blood lead level and expression of T cell subgroup were measured with graphite furnace atomic absorption spectroscopy and immunofluorescence methods respectively. Student t test was used in data analysis, and linear correlation analysis was used to assess the correlation between blood lead level and expression of T cell subgroup.level and expression of T cell subgroup.pression of CD3 and ratio of CD4 to CD8 were lower in Group 1 (lead level ≥0.48 μmol/L) than Group 3 (lead level < 0.24 μmol/L) (t=3.27,blood lead level showed had significant inverse correlation with CD3 expression and the ratio of CD4 to CD8(r=-0.689,-0.674,P < 0.01).CONCLUSION: A blood lead level ≥ 0.48 μmol/L, is shown to significantly decrease the expression of CD3 and ratio of CD4 to CD8 in peripheral blood, which may impair the children's immunological function.
5.Influence of blood lead level on the expression of T cell subgroup in children
Shiguang LI ; Kui FU ; Chunru QI ; Xianguo WU ; Qinghao KONG ; Weiping XI ; Yingyi LIANG
Chinese Journal of Tissue Engineering Research 2005;9(43):170-171
BACKGROUND: Although we have gained much information about leadinduced organ damage, the effect of blood lead level on T cell subgroup is yet to be determined.OBJECTIVE: To assess the effect of blood lead level on T cell subgroup of children, and the association of T cell subgroup with threshold limit value of blood lead level.DESIGN: Comparative observation.SETTING: The Institute of Molecular Microorganism, Maternity and Child Care Hospital, Shanxi Provincial Children Hospital.PARTICIPANTS: Sixty children, (32 boys and 28 girls), aged 3-6 years with a mean of (4.5±1.5) years old, were selected from the suburbs of Taiyuan city between May 2003 and October 2003. Informed consents were obtained from all their guardians. The enrolled children according to their blood lead levels were assigned into three groups, 13 in Group Ⅰ with blood lead level ≥ 0.48 μmol/L, 20 in Group 2 with lead level ≥ 0.24 μmol/L but < 0.48 μmoL/L and 27 in Group 3 with lead level < 0.24 μmol/L.METHODS: Blood lead level and expression of T cell subgroup were measured with graphite furnace atomic absorption spectroscopy and immunofluorescence methods respectively. Student t test was used in data analysis, and linear correlation analysis was used to assess the correlation between blood lead level and expression of T cell subgroup.level and expression of T cell subgroup.pression of CD3 and ratio of CD4 to CD8 were lower in Group 1 (lead level ≥0.48 μmol/L) than Group 3 (lead level < 0.24 μmol/L) (t=3.27,blood lead level showed had significant inverse correlation with CD3 expression and the ratio of CD4 to CD8(r=-0.689,-0.674,P < 0.01).CONCLUSION: A blood lead level ≥ 0.48 μmol/L, is shown to significantly decrease the expression of CD3 and ratio of CD4 to CD8 in peripheral blood, which may impair the children's immunological function.
6.Anterior decompression and fusion with n-HA/PA66 cage for the treatment of lower cervical fracture and dislocation.
Xi YANG ; Yue-Ming SONG ; Li-Min LIU ; Chao-Liang LÜ ; Qing-Quan KONG ; Chong-Qi TU
China Journal of Orthopaedics and Traumatology 2014;27(2):92-96
OBJECTIVETo explore the clinical effects of anterior decompression and fusion with a nano-hydroxyapatite/ polyamide 66 (n-HA/PA66) cage in treating lower cervical fracture and dislocation.
METHODSFrom January 2008 to December 2010, the clinical data of 42 patients with lower cervical fracture and dislocation were retrospectively analyzed. There were 29 males and 13 females aged from 20 to 65 years old. The mean age was 46.8 years. Five cases got injuried in C3, 14 cases in C4, 12 cases in C5, 7 cases in C6 and 4 cases in C7. According to Frankel grade, 4 cases were classified in grade A, 11 cases in grade B, 13 cases in grade C, 9 cases in grade D and 5 cases in grade F. Twenty-eight cases were treated with anterior corpectomy and fusion and 14 cases with anterior discectomy and fusion. Frankel grade was used to do neurologic assessment and visual analogue scale (VAS) was used to evaluate the improvement of clinical symptoms. Segmental height and sagittal lordosis were measured by radiographs and cage location. Cage appearance and fusion status were assessed by 3D-CT images.
RESULTSAll patients were followed up for 3 to 5.2 years with an average of 4.1 years. Frankel grade had obviously improved than preoperative (Z = -4.845, P < 0.001). There were 2, 3, 11, 8, 11 cases classified in grade A, grade B, grade C, grade D and grade E respectively. At the third day after operation and latest follow-up,VAS was (2.6 +/- 1.8),(1.3 +/- 1.0) scores respectively. Both had improved than preoperative (P < 0.05). Up to the latest follow-up, there was only one patient (2.4%) with slight cage translocation (less than 2 mm), however, no cage prolapsed, or collapse, or breakage were found. Both segmental height and lordosis improved significantly after surgery (P < 0.001). And there was not significant difference in both parameters between each postoperative time points (P > 0.05). The mean distance of cage subsidence was 1.5 mm and the rate of cage subsidence (> 3 mm) was 4.8%.
CONCLUSIONThe n-HA/PA66 cage can not only restore and maintain the fusion segmental height and radian, but also promote the osseous fusion and profit the radiographic assessment after operation. Thus, it was an ideal material for prop graft.
Adult ; Aged ; Cervical Vertebrae ; injuries ; surgery ; Decompression, Surgical ; methods ; Female ; Humans ; Joint Dislocations ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fractures ; surgery ; Spinal Fusion ; methods
7.Effects of Chinese herbal compound on monoamine and neuronal amino acids in rat's telencephalon in the course of exhaustion and recovery process.
Hong-Zhen LIU ; Li ZENG ; Xi-Liang KONG ; Iei ZHU ; Yun-Chao MA
Chinese Journal of Applied Physiology 2011;27(4):439-443
OBJECTIVETo observe the effect of Chinese herbal compound on variance of neurotransmitters in rat telencephalon and to further discuss the mechanism underlying Chinese herbal compound in improving exercise capacity and promoting recovery from exercise-induced fatigue.
METHODS64 rats (8 week old) were randomly divided into medicine group (MG) and control group (CG). Chinese herbal compound was administered to rats of MG for 8 weeks. 8 weeks later, every group was divided into 4 subgroups and all were killed at different time point separately, and then neurotransmitter in rat brain was tested.
RESULTSThe exhaustion time of MG was significantly longer than that in CG (P < 0.01). In rest conditions, glutamic acid (GLU) of MG was significantly higher than that in CG (P < 0.01), while, there were no significant differences between MG and CG in other indexes. After fixed quantitative load exercise, the content of 5-hydroxytryptamineZZ(5-HT), 5-hydroindole acetic (5-HIAA), gamma-aminobutyric acid (GABA), Dopamine (DA) and 5-HT/5-HIAA were significantly lower than those in CG, while, GLU, GLU/GABA and DA/5-HT were significantly higher than those in CG. Compared with CG, exhaustion significantly (P < 0.05) decreased 5-HT, GABA and 5-HT/5-HIAA, and significantly (P<0.05) increased GLU, DA/5-HT and GLU/GABA level in MG. 12 h after exhaustion, in contrast to CG, level of 5-HT and 5-HT/5-HIAA in MG were significantly (P < 0.01) lower while GLU, DA, GABA and DA/5-HT were significantly (P < 0.01) higher.
CONCLUSIONDuring exhaustion exercise, Chinese herbal compound demonstrated strong inhibiting effect on synthesis of 5-HT, 5-HIAA, DA, GABA and promoting effect on GLU synthesis, this had been confirmed by the combined effect, including increase of excitatory transmitter and excitability of central nervous system and the prolongation of exhaustion time and promoting recovery from fatigue.
Amino Acids ; metabolism ; Animals ; Biogenic Monoamines ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Fatigue ; metabolism ; physiopathology ; Hydroxyindoleacetic Acid ; metabolism ; Male ; Neurons ; metabolism ; Physical Conditioning, Animal ; physiology ; Physical Exertion ; physiology ; Rats ; Rats, Wistar ; Serotonin ; metabolism ; Telencephalon ; drug effects ; metabolism
8.Study on the efficacy and safety of Xueyou Mixture in treating hemophilia.
Shan-xi LIU ; Lin JIANG ; Yi-guo LIU ; Yue-qin HE ; Xin LIANG ; Wei-wei KONG ; Jing CHEN
Chinese journal of integrative medicine 2007;13(2):141-144
OBJECTIVETo observe the effect of Xueyou Mixture (, XYM) on blood coagulation factors and its safety in treating hemophilia.
METHODSTo the randomly selected 65 inpatients of hemophilia, XYM was administered accompanied with intravenous dripping of liver cell growth factor 60-100 mg once a day to protect the liver, with no blood products like concentrated VIII and FIX factors or blood plasma given. The treatment lasted for 3 weeks. The short-term efficacy and adverse reactions were observed. The long-term efficacy in patients was observed in a follow-up study of 6-12 months after they were discharged from the hospital but continuously took XYM orally.
RESULTSThe short-term markedly effective rate in the patients was 95.38% (62/65). After they were treated for 3 weeks, the level of FVIII factor activity increased in 56 patients of type A from (3.32+/-2.21) % to (4.18+/-2.23) %, and in 9 of type B from (4.92+/-1.81) % to (5.64+/-1.96) %. Compared with that before treatment, the difference was significant in both of them (P<0.01). No obvious adverse reaction was found in the treatment period. The follow-up study showed that in 22 patients of type A, the FVIII factor activity ratio increased from (3.25+/-2.11) % to (6.31+/-2.16) %, (8.36+/-1.05) %, and (16.38+/-2.71) % in the 2nd, 3rd and 6th month after discharge respectively, all showing significant difference to that before treatment (P<0.01); and in 4 patients of type B, it increased from (4.15+/-2.26) % to 7.8% and 11.6% (mean value) in the 2nd and 6th month respectively.
CONCLUSIONXYM could raise the activity of factors VIII and IX in patients with hemophilia, and the degree of the rise is related with the duration of the therapy, with no obvious adverse reaction, which strikes out a new path and new train of thinking for the treatment of the disease by nonblood preparation.
Adolescent ; Adult ; Blood Coagulation Factors ; analysis ; Child ; Child, Preschool ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Follow-Up Studies ; Hemoglobins ; analysis ; Hemophilia A ; blood ; drug therapy ; Humans ; Infant ; Male ; Medicine, Chinese Traditional ; Middle Aged
9.Strategy for the diagnosis and treatment of bilateral testicular tumor
Da-Xin GONG ; Zhen-Hua LI ; Ze-Liang LI ; Xia WANG ; Shao-Bo YANG ; Jian-Bin BI ; Gang LI ; Zhi-xi SUN ; Chui-ze KONG
Chinese Journal of Urology 2001;0(10):-
Objective To evaluate the clinical features and the strategy for the diagnosis and treat- ment of bilateral testicular tumor.Methods The clinical data (including the signs and symptoms,imaging studies,tumor markers,treatment modalities and histopatbologic diagnoses) of 10 cases of bilateral testicular tumor from January 1980 to December 2004 were reviewed.Their age ranged from 19 to 58 years(mean,34 years).Of the 10 cases,8 with metachronous and 2 with synchronous testicular tumors were identified.The clinical stages at the primary and secondary tumor diagnosis were:5 cases of stageⅠ,3 of stageⅡ;and 6 cases of stageⅠ,1 of stageⅡ,and 1 of stageⅢ,respectively,in 8 metachronous tumor patients.Two syn- chronous tumor patients were both identified as stageⅠdisease.Histological examination showed the primary tumor (seminoma) in 4 cases and the secondary contralateral tumor (seminoma) in 3.Results Two syn- chronous tumor patients underwent bilateral radical orchiectomy simultaneously,and 8 underwent orchiectomy successively.Retroperitoneal lymph node dissection was performed in 3 cases.Postoperatively,hypogonadism occurred in 10 patients,and 7 of them received androgen replacement therapy.Follow-up ranged from 9 month to 23 years with a mean of 10.5 years.Two patients died of the disease;2 had metastasis (1 of them was alive with metastasis);2 had recurrences and underwent local resection.Conclusions Metachronous bilateral testicular cancers are more common than synchronous bilateral testicular cancers.Seminoma was the most common histopathologic type.Testis-sparing surgery can be performed in selected cases.
10.A preliminary study on the expression and biological function of recombinant human SCF-TPO fusion protein.
Nan LIU ; Yong-zhi XI ; Si-qi GUO ; Yu-ying SUN ; Zhi-hong YUAN ; Jian-wu CUI ; Cai-xia XI ; Fei LIANG ; Fan-hua KONG
Chinese Journal of Hematology 2005;26(1):19-22
OBJECTIVETo study the expression of recombinant human SCF-TPO fusion protein and its biological function.
METHODSFour primers were designed according to known sequences of TPO and SCF. The functional amino acid domains of TPO and SCF were amplified by RT-PCR from fetus hepatocytes, respectively. The expression plasmid pET32a/SCF-TPO was constructed by VOE gene fusion technique and expressed in E. coli BL21(DE3) plysS as inclusion body after isopropyl-beta-D-1-thiogalactopyranoside (IPTG) induction. The fusion protein was tested by SDS-PAGE and Western blot. The biological functions of SCF-TPO fusion protein in MO7e cells was investigated by MTT method after purification with metal chelating chromatography.
RESULTSThe high expression SCF-TPO fusion protein was obtained, reaching up to 30% of the total cellular protein. Western blot verified the correct fusion expression and MTT results showed the growth promoting effect of the SCF-TPO fusion protein on MO7e cells, with a higher promoting activity at 100 ng/ml.
CONCLUSIONSExpressed SCF-TPO fusion protein after renaturation has biological activity in promoting the proliferation of MO7e cells.
Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; genetics ; physiology ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Genetic Vectors ; Humans ; Recombinant Fusion Proteins ; genetics ; metabolism ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cell Factor ; genetics ; metabolism ; physiology ; Thrombopoietin ; genetics ; metabolism ; physiology