Therapeutic effect of Panax notoginseng saponins combined with cyclophosphamide in mice bearing hepatocellular carcinoma H22 cell xenograft / 南方医科大学学报

OBJECTIVE@#To investigate the therapeutic effects of total saponins from Panax notognseng (PNS) combined with cyclophosphamide (CTX) in mice bearing hepatocellular carcinoma H22 cell xenograft.@*METHODS@#We examined the effects of treatment with different concentrations of PNS on H22 cell proliferation for 24 to 72 h in vitro using CCK8 colorimetric assay. Annexin V/PI double fluorescence staining was used to detect the effect of PNS on apoptosis of H22 cells. Mouse models bearing H22 cell xenograft were established and treated with CTX (25 mg/kg), PNS (120, 240 or 480 mg/kg), alone or in combinations. After treatments for consecutive 10 days, the mice were euthanized for examinations of carbon clearance ability of the monocytes and macrophages, splenic lymphocyte proliferation, tumor necrosis factor (TNF-α), interleukin-2 (IL-2), serum hemolysin antibody level, blood indicators, and the tumor inhibition rate.@*RESULTS@#Treatment with PNS concentration-dependently inhibited the proliferation and significantly promoted apoptosis of cultured H22 cells (P < 0.01). In the tumor-bearing mouse models, PNS alone and its combination with CTX both resulted in obvious enhancement of phagocytosis of the monocyte-macrophages, stimulated the proliferation of splenic lymphocytes, promoted the release of TNF-α and IL-2 and the production of serum hemolysin antibody, and increased the number of white blood cells, red blood cells and lymphocytes in the peripheral blood. Treatment with 480 mg/kg PNS combined with CTX resulted in a tumor inhibition rate of 83.28% (P < 0.01) and a life prolonging rate of 131.25% in the mouse models (P < 0.05).@*CONCLUSION@#PNS alone or in combination with CTX can improve the immunity and tumor inhibition rate and prolong the survival time of H22 tumor-bearing mice.

Electrochemical Immunosensor for Microcystin-LR Detection Based on Nano-composite Material Immobilization and Enzymatic Amplification / 分析化学

A competitive electrochemical immunosensor based on the nano-composite material immobilization and enzymatic amplification was designed for detection of microcystin-LR. Gold nanoparticles/carboxylated multi-walled carbon nanotubes (AuNPs/c-MWCNTs) composite film, which formed by electrodepositing of AuNPs on the C-MWCNTs modified glassy carbon electrode,was used for the immobilization of the antibody of microcystin-LR (anti-MCLR). Horseradish peroxidase (HRP) was introduced onto the nanocomposite interface by HRP blocked sensing interface and specific capture of antibody with target. It could be employed to catalyze the reduction of H2O2, and to block the possible remaining active sites as well. A competitive immunoreaction between antigen and MCLR-HRP was used for target analysis. In the presence of H2O2and hydroquinone (HQ),MCLR could be indirectly detected with differential pulse voltammetry (DPV) method by determining the reduction current of HQ. Under the optimal conditions, the proposed immunosensor exhibited wide linear ranges in the concentration ranges of 0.1-100.0 μg/L, with a detection limit of 0.038 μg/L (S/N = 3,n=8). The immunosensor showed good specificity, stability and sensitivity. It was used to determine MCLR in real water samples with the recoveries of 72.9%-117.3%.

Effects of Strengthened Core Stability Training with Band on Motor Function and Balance in Children with Spastic Cerebral Palsy / 中国康复理论与实践

Objective To investigate the application of band to strengthen core stability training for cerebral palsy. Methods From May, 2015 to December, 2016, 70 children with spastic cerebral palsy in outpatient department were di-vided into control group (n=35) and observation group (n=35). Both groups accepted routine rehabilitation train-ing, and the control group accepted core stability training, while the observation group was trained with a band during core stability training, for twelve weeks. They were assessed with Gross Motor Function Measure- 88 (GMFM-88), Berg Balance Scale (BBS), Manual Muscle Test (MMT) before and after treatment. Results The scores of GMFM- 88, BBS and MMT of external oblique improved in both groups after treatment (t>12.904, P<0.001), and improved more in the observation group than in the control group (t>2.121, P<0.05). Conclusion Strengthening core stability training with a band can further improve gross motor function, balance and mus-cle strength in the spastic cerebral palsy children.

Clinical analysis of 10 cases of pediatric Crohn's disease / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the clinical features and treatment of pediatric Crohn's disease (CD).</p><p><b>METHODS</b>Clinical data of 10 children with active CD diagnosed between 2005 and 2013 were retrospectively reviewed.</p><p><b>RESULTS</b>Abdominal pain, diarrhea, and bloody stools were the most common symptoms in these patients, usually accompanied by different degrees of growth retardation and nutritional disorders. Fever was the main extraintestinal manifestation. Enteroscopy showed discontinuous and segmental mucosal hyperaemia and erosion, cobblestone appearance and mucosal ulceration. Abdominal ultrasound revealed uneven and segmental thickening of the intestinal wall. The pathological esamination showed many lymphocytes, eosinophils and plasma cells infiltrating into the lamina propria and partial atrophy of mucosal gland. C-reactive protein (CRP) level was significantly lower in the remission stage than in the acute stage and the recurrence stage (P<0.05). The erythrocyte sedimentation rate (ESR) was significantly lower in the remission stage than in the recurrence stage (P<0.05). Among mild cases identified by the pediatric Crohn's disease activity index (PCDAI) in the early stage of disease, the induced remission rate and maintained remission rate were 100% and 67%, respectively, with oral 5-aminosalicylic acid (5-ASA) and adrenocortical hormone. Among moderate and severe cases identified by the PCDAI, the partial remission rate was 100% with 5-ASA and adrenocortical hormone, but the maintained remission rate was not so good and the recurrence rate of disease was high.</p><p><b>CONCLUSIONS</b>Pediatric CD has no specific clinical manifestations and laboratory test results. ESR and CRP can be used as the markers for evaluating the disease progression. 5-ASA has certain efficacy in inducing and maintaining remission of pediatric CD. There is a certain correlation between treatment outcome and the PCDAI score in the early stage of disease.</p>

Gene sequence analysis and prokaryotic expression of sigmaB protein of Muscovy duck reovirus YB strain / 病毒学报

Muscovy ducks reovirus (DRV) is an important pathogen with a high mortality rate in Muscovy ducks, the researches in the test and the immunity were useful for the prevention and control of DRV infection. In this study, the S3 genes of the three Fujian DRVs were cloned by RT-PCR and sequencing technology. It was found that DRV-YH and YJL were close to avian reovirus (ARV) in the genetic distance, with high identities ranged from 94. 6% to 98. 9%, however, the identities of DRV-YB strain and reference ARV strains in the S3 gene were only 60.6% - 61.7%. The expression vector pET-30a-S3 harboring DRV YB strain S3 gene was constructed and transformed into E. coli BL21, and then the fusion sigmaB protein expression was induced with IPTG. The SDS-PAGE of the expressed products indicated that the fusion protein of approximately 42ku in molecular weight was expressed highly in inclusion body, and made up 67. 7% of the total proteins. The most efficient concentration of IPTG and inducing time were 0. 1 mM and 5h respectively, while the best temperature for expression was 37 degrees C. After purification with the Ni2+ affinity chromatography, the fusion sigmaB protein was 93% of the total proteins, and the purified protein amounted to 0. 86g/L. The Western blot analysis showed that the fusion aB protein was recognized specifically by the antiserum against DRV, confirming that the recombinant fusion protein had good immunoreactivity.

Valproic acid-induced idiosyncratic liver injury in 4 cases / 中华儿科杂志

<p><b>OBJECTIVE</b>Children with refractory epilepsy who suffered from severe liver function impairment during valproic acid (VPA) treatment at routine dosage were studied. The clinical manifestations and therapeutic approaches were investigated in order to improve its diagnosis and management.</p><p><b>METHOD</b>Clinical information as well as features and management of 4 inpatients who were suffered from intractable epilepsy with severe liver function impairment induced by VPA since 2006 were collected and analyzed, including age of onset of epilepsy, VPA using age and the time when liver injury occurred, clinical manifestations, auxiliary examinations and management.</p><p><b>RESULT</b>Among the 4 cases, three were male and one was female. The admitted age ranged from 1 - 9 years and 1 month. The course of disease was 25 d - 6 months. They manifested as refractory epilepsy of epilepsia partialis continua which was difficult to control. After using VPA for 62 d (50 - 76 d), all developed severe impairment of liver synthetic function which was not related to the concentration of VPA. One was diagnosed with Alpers syndrome, two were suspicious of Alpers syndrome, and the other was diagnosed gliocytoma after brain biopsy. VPA was stopped immediately and symptomatic therapies were used. Other than that, intravenous injection of L-carnitine in 3 cases recovered the liver function.</p><p><b>CONCLUSION</b>VPA-associated severe hepatotoxicity can manifest first as impaired liver synthetic function. Besides alanin transaminase and aspartate transaminase, the liver synthetic function test is more important than monitoring of liver enzymatic functions in monitoring for the hepatotoxicity. Intravenous injection of L-carnitine in early stage showed good treatment effect.</p>

Changes of cholesterol 7alpha-hydroxylase, farnesoid X receptor, and small heterodimer partner expression in liver tissues of rats with obstructive cholestasis / 第二军医大学学报

Objective: To investigate the changes of cholesterol 7alpha- hydroxylase(CYP7A1) ,farnesoid X receptor(FXR), and small heterodimer partner (SHP) expression in liver tissues of bile duct ligated (BDL) rats,and to analyze the relationship between their expression. Methods: Obstructive cholestasis rat model was induced by bile duct ligation,and rats were sacrificed on day 1,3,and 14 after operation. The hepatic tissues were collected and the total mRNA was extracted. The CYP7A1 mRNA expression was determined by real-time RT-PCR. The protein expression of FXR and SHP in the nuclei was determined by Western blotting analysis. Results: Compared with sham-operated group, experimental obstructive cholestasis group had significantly enhanced CYP7A1 mRNA expression and decreased FXR,SHP expression(P<0. 001,P<0. 05). Conclusion: The up-regulation of CYP7A1 may be associated with the down-regulation of FXR and SHP.

Mechanism of inhibitory effect of P7 on 3T3 cell proliferation induced by basic fibroblast growth factor / 药学学报

To investigate the mechanism of inhibitory effect of a novel bFGF antagonist peptide isolated from the phage display random heptapeptide library on cell proliferation induced by basic fibroblast growth factor. The effect of P7 on cell morphology was observed under an inverted microscope. Flow cytometry was applied to analyze the effect of P7 on cell cycle progress of bFGF-stimulated cells. The effect of P7 on bFGF-induced activation of MEK and Erk1/2 in MAPK pathway was detected by Western blotting. The results showed that no significant cell morphology change was observed in the range of detected concentrations of P7. Cell cycle analysis showed that P7 decreased S-phase cell population and arrested cell cycle at the G0/G1 phase of bFGF-stimulated cells. The results of MAP kinase activation assay indicated that P7 decreased bFGF-induced MEK and Erk1/2 phosphorylation in a dose-dependent manner. P7 inhibited proliferation of bFGF-stimulated Balb/c 3T3 cells possibly via cell cycle arrest at the G0/G1 phase and down-regulation of signal molecular activation in MAPK pathway.

Proteomic study of the effect of recombinant hFGF-10 adenovirus on HaCat cells / 药学学报

This study is to investigate the effect of recombinant hFGF-10 adenovirus on the proteome of HaCat cells, and to speculate further the possible mechanism of the effect of hFGF-10 on HaCat cells via differentially expressed proteins identified. Two-dimensional gel electrophoresis (2-DE) combined with tandem time-of-flight mass spectrometry was applied to identify the differentially expressed protein spots on the 2-DE maps of the whole-cell proteins from Ad infected and rAd-hFGF-10 infected HaCat cells. The mRNA and protein levels of the differentially expressed proteins were confirmed with semi-quantitative RT-PCR and Western blotting. The results showed that the 2-DE maps with high resolution were obtained, and four selected differentially expressed proteins involved in cell apoptosis, cytoskeleton regulation and protein degradation were identified with MALDI-TOF/TOF. The mRNA and protein levels of one of the differentially expressed proteins, VDAC2, were up-regulated in HaCat cells infected with the recombinant hFGF-10 adenovirus. The differentially expressed protein, VDAC2, may be related to the bioactivities of hFGF-10.

Studies about the level of CD4+ CD25+ regulatory T cells and relation between expression of Foxp3 and CD127 in peripheral blood of chronic HBV infection / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the level of CD4+ CD25+ Foxp3+ regulatory T cells and observe relation between expression of Foxp3 and CD127 in peripheral blood of chronic HBV infection.</p><p><b>METHODS</b>CD4+ CD25+ Foxp3+ and CD4+ CD25+ CD127low Treg in peripheral blood from 34 patients of immune tolerance stage, 26 patients of immune clearance stage and 31 patients of non-active status were quantitatively analyzed by flow cytometry.</p><p><b>RESULTS</b>Immune tolerance group presented a higher fraction of CD4+ CD25+ Foxp3+ and CD4+ CD25+ CD127low Treg than non-active group in chronic HBV infection (Z = -2.693, P = 0.007 and t = 3.251, P = 0.002), and HBV positive group also presented a higher fraction than non-active group (t = 2.266, P = 0.026 and t = 3.208, P = 0.002), But ALT normal group is similar to ALT abnormal group (P > 0.05). In this study, the relation between expression of CD127low and Foxp3+ from CD4+ CD25+ regulatory T cells was observed, and CD4+ CD25+ CD127low Treg presented a higher fraction than CD4+ CD25+ Foxp3+ Treg.</p><p><b>CONCLUSION</b>Peripheral Treg in HBV active replication group is higher than HBV negative group of chronic HBV infection. Expression of CD127low is consistent with Foxp3+ in CD4+ CD25+ regulatory T cells, but the former is significantly higher than the latter.</p>

Proton magnetic resonance spectroscopy of the thalamus and hypothalamus in patients with first-episode depression / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the presence of abnormal metabolism in the thalamus and hypothalamus in patients with first-episode depression.</p><p><b>METHODS</b>Thirty drug-naive patients with first-episode depression and 30 age-matched controls were scanned with proton magnetic resonance spectroscopy ((1)H-MRS) for Naa, Cho, Cr and mI.</p><p><b>RESULTS</b>Compared with the control group, the patients showed significantly reduced mI and mI/Cr of the hypothalamus, reduced mI/Cr of the left thalamus, and lowered Cho, ml, and ml/Cr of the right thalamus (P<0.05).</p><p><b>CONCLUSION</b>Patients with first-episode depression may have myo-inositol and phosphoric acid metabolism disorder in the thalamus and hypothalamus with malfunction of cellular osmotic pressure adjustment mechanism. Abnormal mI/Cr in the thalamus and hypothalamus may represent an important biochemical change in advanced patients with depression.</p>

Effects of different die materials on fracture and failure behaviors of all-ceramic crowns / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the influence of different die materials on fracture strength and failure mode of IPS e.max Press crowns in mechanical test.</p><p><b>METHODS</b>Thirty-six IPS e.max Press crowns were divided into 3 groups. They were cemented to extracted maxillary first premolars in Group 1, to epoxy dies in Group 2, and to porous NiTi alloy dies in Group 3 respectively. All the crowns were subjected to compressive load in an universal mechanical testing machine. The maximum fracture strength(N) was recorded and fracture modes of the tested crowns were classified. Fractographies of the failed crowns were observed with a stereomicroscope.</p><p><b>RESULTS</b>Statistical analysis results showed that the mean fracture strength of the IPS e.max Press crowns on extracted natural teeth [(1159.7 +/- 256.0) N] and porous NiTi alloy dies [(1229.6 +/- 326.7) N] were obviously higher than that of epoxy dies [(883.5 +/- 198.1) N, P = 0.016 and 0.003]. Observations of the fractographies of failed crowns showed that the crowns on epoxy die fractured mesio-distally and no plastic deformation and characteristic cracks were observed. However, crowns in the other groups showed cone cracks at the contact area on occlusal surface and radical cracks initiated from the adhesive interfaces.</p><p><b>CONCLUSIONS</b>Die material has a significant influence on fracture resistance and fracture mode of the IPS e.max Press crowns. The crowns cemented to porous NiTi alloy dies show a similar fracture mode as those on extracted natural teeth.</p>

Establishment and optimization of two-dimensional gel electrophoresis technique of proteome from bone marrow leukemia cells / 中国实验血液学杂志

The aim of this study was to establish and optimize two-dimensional electrophoresis method for human bone marrow leukemia cells in order to obtain the profiles with high resolution and reproducibility. The total protein was extracted and separated by isoelectric focusing (IEF) and SDS polyacrylamide gel electrophoresis (SDS-PAGE). The gels were stained with silver nitrate or Coomassie brilliant blue, and then scanned and analyzed with PDQuest 7.4 analysis software. The effects of different protein preparation methods and electrophoresis conditions on the profiles were compared. The results indicated that by optimizing preparation of protein sample and electrophoresis protocols, clear profiles with 780 +/- 73 well separated protein spots on an average were obtained and the match rate was 82 +/- 5% between reproducible gels from leukemia cells of different sub-type. It is concluded that the two-dimensional electrophoresis method of proteome from human bone marrow leukemia cells is established successfully and is suitable for the further comparative proteomic research between leukemia of different types.

The immunostimulatory study on peripheral blood mononuclear cell and CD4+ T lymphocyte of HBV infections that were activated by CpG oligonucleotide / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the effects of CpG oligodeoxyribonucleotide (ODN) as adjuvant on the immune responses in PBMC and CD4+ T cell with chronic hepatitis B virus.</p><p><b>METHODS</b>The selected 20 infections were averagely divided two groups. The frequency of IFN-gamma secreting PBMC and CD4+ T cell in immune tolerant phase and in the immune clearance phase that had stimulated by CpG ODN, HBsAg and Mixture [CpG ODN + HBsAg] were analyzed by enzyme linked immune spot (ELISOT).</p><p><b>RESULTS</b>The PBMC and CD4+ T cell were differently incubated by CpG ODN, HBsAg and M [CpG ODN + HBsAg]. The number of IFN-gamma spot differently are 3 +/- 8, 339 +/- 429, 375 +/- 496, 1 +/- 4, 5 +/- 16 and 5 +/- 12; the results of immume tolerance are 3 +/- 8, 361 +/- 153, 375 +/- 276, 0 +/- 2, 2 +/- 2 and 4 +/- 4; but the results of immune clearance are 3 +/- 21, 289 +/- 345, 405 +/- 656, 2 +/- 14, 8 +/- 40 and 7 +/- 30. The IFN-gamma spots statistical analysis of PBMC were differently incubated by HBsAg and M, the total is P = 0.720, The IFN-gamma spots statistical analysis of CD4+ T cell were differently incubated by HBsAg and M, the total is P = 0.890, The IFN-gamma spots statistical analysis of PBMC and CD4+ T cell were differently incubated by M, the total is P = 0.000.</p><p><b>CONCLUSIONS</b>The ability that CpG ODN can not significantly increase the IFN-gamma secreting of PBMC and CD4+ T cell that were incubated by HBsAg to the infection in immune tolerant phase and in the immune clearance phase, but the PBMC outweighed The CD4 T cell.</p>

Clinical observation of gefitinib in treatment of female patients with adenocarcinoma of lung WU / 肿瘤研究与临床

Objective To evaluate the efficacy and adverse effects of gefitinib in the treatment of fe- male patients with advanced adenocarcinoma of lung who had failed to previous chemotherapy.Methods These patients received 250mg of gefitinib orally,once daily until disease progression or development of intol- erable toxic reaction.They were evaluated one month after treatment and every other month thereafter.Results Among the 27 evaluable patients,there were 1 CR(3.7%),11 PR(40.8%),10 SD(37.0%)and 5 PD(18.5%). The overall response rate was 44.5%(95% CI 29%～68%);and 22 patients(81.5%)gained profit(CR+PR+ SD)from the clinical therapy(95% CI 62%～94%);the mean TTP was 7.2 months.Symptomatic improvement rate was 80.0%.The main adverse effects were mild rash and diarrhea.Conclusion gefitinib has significant efficacy in the treatment of female patients with advanced tung cancer who had failed to previous chemother- apy.Adverse effects are mild.gefitinib is a suitable therapy for these patients.

Recombinant Adenovirus Infected HaCat Cells Express Human Kerotinocyte Growth Factor (hKGF) / 中国生物工程杂志

Human kerotinocyte growth factor (hKGF) gene amplified by PCR was inserted into the shuttle vector pAdTrack-CMV to get the recombinant plasmid pAdTrack-CMV-hKGF, which was linearized with Pme I and transferred into Escherichia coli BJ5183 containing the adenoviral bone plasmid pAdEasy-1 to obtain the recombinant adenoviral plasmid pAdEasy-hKGF. The recombinant adenoviral plasmid was then transfected into HEK-293 cell lines via Lipofectamine 2000 to package and amplify the recombinant adenovirus containing hKGF gene detected by PCR. The recombinant adenovirus produced could effectively infect HaCat cells. The result of Western blotting showed that HaCat cells infected with the recombinant adenovirus expressed and secreted hKGF protein.

Selection of bFGF Mimic Peptide by Phage Display / 中国生物工程杂志

Objective: To obtain the bFGF mimic peptide binding to FGFR via phage display, and to provide the base for developing peptide agonist of bFGF. Methods: Using Balb/c 3T3 cells as the target cells and COS-7 cells as the subtractive panning, the phage display heptapeptide library was biopanned for 4 rounds to obtain the single phage clones. The affinity and the specificity of the clones were assessed by ELISA. DNA sequencing was applied to further analyze the positive clones. Results: Twelve positive clones were selected from the enriched phages. A group of hydrophobic peptides containing a conserved motif, PR, was identified. Conclusion: Two bFGF mimic heptapeptides binding to FGFR were selected, which may be used as the candidates for bFGF agonist.

Effect and mechanism of action of non-mitogenetic human acidic fibroblast growth factor on the mitogenic activity of the mammary tumor cell / 药学学报

<p><b>AIM</b>To compare the effects of the non-mitogenetic human acidic fibroblast growth factor (nmhaFGF) and the human acidic fibroblast growth factor (haFGF) on the proliferation and MAPK signal transduction pathway of the malignant tumor cell and to study the clinical safety of nmhaFGF.</p><p><b>METHODS</b>The mammary tumor cells (MCF-7) were treated with haFGF and nmhaFGF separately. The mitogenic activities of both haFGF and nmhaFGF were detected by MTT method and the cell cycle was analyzed by flow cytometer (FCM). The expression levels of the signal proteins, Grb2 (growth factor receptor bound 2) and ERK1/2 (extracellular signal-regulated kinase 1/2), were detected by semi-quantitative Western blotting method.</p><p><b>RESULTS</b>The mitogenic activity of nmhaFGF was obviously lower than that of haFGF. The activity of nmhaFGF was weaker than that of the haFGF. The ratio of G1/G0, G2/M of haFGF was markedly lower than that of nmhaFGF and control group, and was reverse in S phase. The expression levels of both Grb2 and ERK1/2 of the nmhaFGF treated group were lower than that of the haFGF treated group and approaching the control group.</p><p><b>CONCLUSION</b>The mitogenic activity of the nmhaFGF decreased remarkably. Its mechanism probably via down-regulation of the expression of the signal moleculars, MAPK-ERK1/2 and Grb2.</p>

Expression and Purification of Basic Fibroblast Growth Factor Mutant with Reduced Mitogenic Activity / 中国生物工程杂志

In order to decrease the potential side-effects of human basic fibroblast growth factor (hbFGF) caused by its broadspectrum mitogenic activity, a single residue of hbFGF, the residue serine 108, was replaced with neutral alanine residue to construct a mutant of hbFGF (mhbFGF) with reduced mitogenic activity. The mutant was overexpressed in Escherichia coli BL21(DE3) by IPTG induction. The expression level of mhbFGF was about 30% of the total cellular protein. The expressed mhbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate. Measured by MTT method, the effect of mhbFGF on Balb/c 3T3 cell proliferation was much lower than that of wild-type hbFGF. The purified recombinant mhbFGF was prepared and sufficient for the following pharmacological and safety studies.

Expression and Purification of Basic Fibroblast Growth Factor Mutant with Reduced Mitogenic Activity / 中国生物工程杂志

In order to decrease the potential side-effects of human basic fibroblast growth factor (hbFGF) caused by its broadspectrum mitogenic activity, a single residue of hbFGF, the residue serine 108, was replaced with neutral alanine residue to construct a mutant of hbFGF (mhbFGF) with reduced mitogenic activity. The mutant was overexpressed in Escherichia coli BL21(DE3) by IPTG induction. The expression level of mhbFGF was about 30% of the total cellular protein. The expressed mhbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate. Measured by MTT method, the effect of mhbFGF on Balb/c 3T3 cell proliferation was much lower than that of wild-type hbFGF. The purified recombinant mhbFGF was prepared and sufficient for the following pharmacological and safety studies.