Objective:To study MUC1 based cancer vaccine.Methods:MUC1 gene was inserted into pMAL-p2 vector and constructed recombinant pMAL-MUC1. MUC1-MBP fusion protein expression was induced by IPTG in E coli DH5? transformed by the recombinant pMAL-MUC1 .The fusion protein was analyzed by Western blot and purified by amylose affinity chromatography. The antiserum,T cell proliferation and CTL activity of spleen from C57 mice immunized by MUC1-MBP were determined respectively by ELISA,adding 3H-TdR and MTT.Results:Had successfully constructed pMAL-MUC1 expression vector,and purified MUC1-MBP and MBP. C57 mice immunized by MUC-MBP generated MUC1 specific antibody and CTL.The titer of polyclonal antibody to MUC1 was about 1∶5 760?3 221. CTL cytotoxicity to the MCF7 and lewis lung cancer cells respectively were at 47.7%?4.3% and 67.5%?6.5%.Conclusion:Human recombinant MUC1-MBP fusion protein activated T and B cell response in mice.The results suggested that the recombinant Muc1 may be used to develop protein vaccine against carcinoma.

Objective: To study the anti-tumor effect of recombinant human MUC1-MBP. Methods: The C57BL/6 mice were in oculated with MUC1-MBP by subcutaneous. MUC1 specific CTL activity of spleen were determined by MTT; The effects on prevention and treatment of tumor were observed by establishing lewis lung cancer-carrying mice. Results: The cytotoxicity of CTL from immunized mice to the MCF7 and Lewis lung cancer cells respectively was (47.7?4.3) % and (67.5 ?6.5) %; 5?10 5 lewis lung cancer cells following immunization were injected iv into C57BL/6 mice, after three weeks, the number of lung and tail tumor colonies was 51 and 5 for PBS and MUC1-MBP groups respectively and the suvival time was significantly delayed in immunized mice. The average volume of tumors in mice with MUC1-MBP was 386 mm 3 wherea control group was 4 000 mm 3 at tumor treating experiment. Conclusions: Recombinant human MUC1-MBP have significantly effects on prevention, treatment and inhibiting metastases of tumor. Our results suggested that the recombinant MUC1-MBP might be used to develop protein vaccine against human carcinoma.

Objective:To transfer human endostatin gene into umbilical cord CD34 + hematopoietic stem cells and detect its expression and excretion. Methods: Human endostatin gene was transferred into human umbilical cord CD34 + hematopoietic stem cells by retroviral pLNCX to build endostatin-transferred cell line.RT-PCR and Western blot analysis were applied to examine the transfection and expression of endostatin gene. Results:RT-PCR proved that genome of endostatin-transferred CD34 + hematopoietic stem cells contained a 550 bp fragment of human endostatin .The expression and excretion of human endostatin from endstatin-transferred hematopoietic stem cells were confirmed by Western blot analysis. Conclusion:Human endostatin gene can be transferred into CD34 + hematopoietic stem cells.

BACKGROUND:The early damaged chondrocytes are susceptible to de-differentiate and exert unstable phenotype during the in vitro culture, thus needing some growth factors.
OBJECTIVE:To observe the promotion effect of insulin-like growth factor 1 on the in vitro proliferation of chondrocytes in adult rabbits with traumatic arthritis.
METHODS:Traumatic arthritis models of adult rabbits were established by using the modified Hulth method. After the models were successful y established, the distal femur and proximal tibia were harvested under sterile conditions, the chondrocytes were cultured. The cultured cells were divided into two groups:control group was cultured with Dulbecco’s modified Eagle’s medium containing 10%fetal bovine serum, while experimental group was cultured with Dulbecco's modified Eagle’s medium containing 100μg/L insulin-like growth factor 1. The effect of insulin-like growth factor 1 on the proliferation of chondrocytes in adult rabbits with traumatic arthritis was determined through the cytomorphology, cellcounting, and cellactivity.
RESULTS AND CONCLUSION:The chondrocytes in adult rabbits with traumatic arthritis were successful y cultured, the majority of cells were mini-cells, presenting smal fusiform, round or polygonal shape. Hematoxylin-eosin staining showed that the number of cells in experimental group was higher than that in control group. MTT assay found that the absorbance of cells in experimental group was greater than that in control group (P<0.01). Our findings indicate that, insulin-like growth factor 1 can promote the in vitro proliferation of chondrocytes in adult rabbits with traumatic arthritis.

Objective:To study the effects of IL-2R? mimic peptide(CP) on skin allograft rejection in mice.Methods:Mouse skin allograft model was employed in all groups.The bioactivities were determined by lymphocyte proliferation,one-way MLR,assay of T lymphocyte subset and the level of cytokine in splenic cell culture supernatant.Results:CP inhibited lymphocyte proliferation;decreased the number of CD4+T lymphocytes in spleen and the value of CD4+/CD8+;down-regulated the level of IL-2 and IFN in splenic cell culture supernatant,as well as prolonged the mean survival times(MST) of skin allografts.The combination of CP and CsA showed cooperativities.Conclusion:CP,as the IL-2R? mimic peptide has the suppressive activities,which can efficiently inhibit skin allograft rejection in mice.

Objective To investigate the relationship between the apparent diffusion coefficient(ADC) value,portal hypertension and hypersplenism.Methods 52 cases underwent MR imaging (including DWI) examination,among them,included normal group(18 cases) hepatic cirrhosis without portal hypertension group(24 cases) and hepatic cirrhosis with portal hypertension group(10 cases).The ADC values of spleen were calculated and compared between groups.The relationship between target of hypersplenism——cells in peripheral blood(RBC/WBC/PLT) and spleen’s ADC values was also analyzed.Results With the appearance of portal hypertension,spleen’s ADC values decreased.The significant difference was found among these 3 groups(P

The use of rnicroarrays of oligonucleotides or cDNA is considered to be a promising approach for DNA and RNA sequence analysis, diagnostics of genetic diseases, gene polymorphism studies and analysis of gene expression. To manufacture cDNA microarrays the samples were printed onto glass microscope slides treated with poly-L-lysine, and then the slides were processed by heat and UV light treatment to attach the cDNA sequence to the glass surface. But the immobilization efficiency of cDNA on the glass surface was low. A simple procedure for manufacture cDNA microarrays on a slide treated with 3-aminopropyltrimethoxysilane is described. The efficiency for attaching cDNA to the amino-modified slides is greater than that to the slides treated with poly-L-lysine. The cDNA microarray made by the amino-modified slides is stable for use in 80℃, 75 % humidity, 3 600Lx light, exposure in air, respectively.

Aim To observe the antifibrogenic effect of resveratrol on mice with schistosomiasis japonica and its effect on Th1 and Th2 responses .Methods Forty-five mice infected with S.japonicum cercariae for 3 weeks were randomly divided into three groups named as infection group ( A) , resveratrol group ( B) and praz-iquantel group ( C) .Fifteen normal mice were taken as normal control group ( D) .In the 13th week post-infec-tion, all mice were sacrificed and the liver tissues were removed.Histopathological changes were observed in the liver of all groups .Splenocytes were prepared from spleens of mice with S.japonicum infection and the proportions of Th1 and Th2 cells in T cells were deter-mined by FACS respectively .RT-PCR was used to de-tect the relative IFN-γ,IL-13,TGF-βmRNA levels in liver tissue .Results After treatment , the degrees of liver fibrosis in groups B and C decreased in the 13th week post-infection ( P <0.01 ) .Compared to group A, the proportions of Th1 cells in group B significantly increased ( P<0.05 ) and the proportions of Th 2 cells in group B decreased significantly ( P <0.01 ) .The level of anti-SWA IgG 2 a in group B was significantly higher ( P<0.05) , while the anti-SEA IgG1 level in group B was lower ( P <0.01 ) than that in group A . The hepatic expression of IFN-γmRNA level in group B was higher than that in group A ( P<0.05 ) , and IL-13 ,TGF-βmRNA levels in group B were lower than in group A ( P<0.01 ) .Conclusion Resveratrol has an antifibrogenic effect through upregulating Th 1 cell re-sponse and downregulating Th 2 cell response in mice infected with Schistosoma japonicum.

Objective To study the effects of acupuncture at myofascial trigger points on spastic foot drop and inversion after stroke. Methods From May, 2014 to May, 2016, 50 stroke patients were randomly divided into control group (n=25) and observation group (n=25). Both groups accepted routine rehabilitation, while the observation group accepted acupuncture at myofascial trigger points per day in addi-tion. They were assessed with Visual Analogue Scale (VAS) of pain, modified Ashworth Scale (MAS), range of motion (ROM) of ankle, sim-plified Fugl-Meyer Assessment (FMA) for lower limbs and maximum walking speed (MWS) in ten metres before and six weeks after treat-ment. Results The scores of VAS, MAS, and FMA, the ROM of ankle, and MWS improved after treatment (t>6.845, P<0.001), and im-proved more in the observation group than in the control group (t>5.586, P<0.001). Conclusion Acupuncture at myofascial trigger points can release spasm to reduce foot drop and inversion in patients with stroke.

OBJECTIVE: To explore the antitumor activities of kushen (Sophora flavescens) flavonoids (KS-Fs) in vivo and in vitro. METHODS: Cell proliferation was assayed by using methyl thiazolyl tetrazolium (MTT) method. H22 hepatocellular carcinoma and S180 sarcoma were induced in ICR mice. Lewis lung carcinoma was induced in C57BL/6 mice. H460 and Eca-109 tumor were induced in Balb/c nude mice by injecting 5x10(5) or 5x10(6) tumor cells in the right flank, respectively. RESULTS: KS-Fs could inhibit the growth of a variety of human tumor cell lines (A549, SPC-A-1, NCI-H460, etc.) in vitro. The antitumor efficacies were confirmed in the mice models of H22, S180 and Lewis lung tumors and the nude mice models of human H460 and Eca-109 xenografted tumors. The oral or intravenous maximum tolerated dose of KS-Fs was more than 2.8 g/kg or 750 mg/kg respectively, far more than the oral medial lethal dose of kushen alkaloids (< or = 1.18 g/kg). No adverse reactions were observed. CONCLUSION: These results suggest that KS-Fs or kurarinone may be developed as a novel antitumor agent.