Objective:To prepare and purify recombinant human zona pellucida 3 protein,and to study its immunologic activity.Methods:The E.coli BL21 containing recombinant plasmid pGEX4T-1/ huZP3 was induced to express GST-fusion protein by IPTG.After a series of purification procedure,the purified protein was analyzed by SDS-PAGE.After the mice immunized with rhuZP3,the antibody responses against rhuZP3 were detected by ELISA.Results:The soluble fusion protein was expressed,and purity of rhuZP3 was 95%.Moreover, purity of rhuZP3 could be recognized by anti-human ZP3 in ELISA.Conclusion:The rhuZP3 is obtained through the preparation of prokaryotic expression system and anti-rhuZP3 antibody has immunological activity.

The electromyography of the superior and inferior heads of lateral pterygoid muscle of 15 human subjects were recorded. It has been observed that marked active potential occurred in the superior head of the ipsilateral pterygoid muscle and the inferior head of the contralateral muscle during a one-side molar bite. Marked active Potential appeared in the superior head of the lateral pterygoid muscle during the closing movement of the jaw, whereas it appeared in the inferior head during the opening movement.It has not been confirmed that the lateral pterygoid muscle is an important protractor of the mandible, as only relatively weak active potential was recorded in the two heads of the muscles on both sides during the protraction of the jaw. Very marked active potential has been observed in the ipsilateral prenygoid muscle during the lateral movement of the mandible, while in the contralatenal superior head it occurred only slightly. The. above finding does not correspond to the descriptions found in current text-books.The two heads of the lateral pterygoid can therefonose considered as two functionally and structurally distinct muscles.

Abstract Objective:To obtain recombinant vaccinia viruses expressed HIV-lcn subtype B gag and gag-hIL-2 chimera gene,evaluate theimmune effects in experimental mice by immunization with HIV-1 DNA vaccine,observe the immune adjuvant effect of IL-2.Metbods: HIV0-1gag and gag-hIL-2 chimera gene were inserted downstream of the combined promotor of pJ38 vector respectively and recombinant vaccinia virus-es were selected by plaque essay. Expression products were examined by SDS-PAGE and Western blot.Immune response of immunized Balb/cmice was evaluated by lymphocyte transformation test,CTL quantity change of T lymphocyte of CD4+、CD8+ and the level of serum antibody.Re-sults:Recombinant vaccinia viruses vJ38gag and vJ38gag-IL-2 were obtained and the expression were confirmed by SDS-PAGE and Western-blot. Recombinant vaccinia viruses show good immune effect when immuned with HIV-1 DNA vaccine.The best strategy for immunization is therVV-primed and HIV-1 DNA vaccine-boosted group and threer times of immunization were suggested to get effective results compared withvJ38gag an incressed immune response was induced by vJ38gag-IL-2. Conclusion: HIV-1 Gag protein and IL-2 protein can be expressed invJ38gag and vJ38gag-IL-2 respectively. IL-2 show good effect to ha sued as immune adjuvant.

The characterization and management of oral and maxillofacial spaces infections in diabetic patients were studied in order to determine the pattern of this clinical condition and formulate a management plan.There were 31 cases with average age of 61 years(s=9);the mean hospitalization time was 14 days(s=6);the average fasting blood glucose level on admission was 10.4 mmol/L.Of the 31 patients 20 were multiple-space infections and 11 were single-space infections.13 patients had major complications during admission.Odontogenic infection was the most common cause of the space infections.Streptococcus viridians and staphylococcus aureus were common organisms(5/19,4/19)identified through pus and/or blood cultures.Early surgical incision and drainage,perfect blood glucose control,intravenous antimicrobial therapy,preventing asphyxia and managing major complications are necessary and effective approaches for the management plan.

Objective To study the antileukemia immune response of IL-18 gene transfected dendritic cells(DCs) of chronic myelogeous leukemia(CML).Methods DCs were transfected with IL-18 gene by liposomes in CML.The expression of IL-18 in IL-18 transfected DCs was detected.The percentages of CD80+ and CD86+ cells in IL-18 tranfected DCs were determined by FCM.The proliferation of T cell,NK and specific CTL kill activity induced by IL-18 gene transfected DCs were detected.Results The quantity of IL-18 in IL-18 tranfected DCs was(596?34.1)pg/2?106cells/48 h,while the culture medium of mock-transduced DCs and DCs did not secrete detectable levels of IL-18.The percentages of CD80+ and CD86+ cells in IL-18 tranfected DCs were higher than that in mock-transfected DCs(P

Objective:To express Gag gp120 fusion protein in the recombinant fowlpox virus.Methods:The Gag gp120 gene of HIV 1 was inserted downstream of the combined promoter in pUTAL,an expressing plasmid was constructed.By transfecting the plasmid into CEF cells pre infected with FPV 282E 4 strain,a recombinant fowlpox virus vUTALG was obtained by selecting in the presence of BUdR and subsequent blue plaque screening.Results:Western blot showed that the recombinant virus expressed the Gag gp120 fusion protein in the infected CEF cells.The virus like particles formed by Gag protein were observed under electron microscope.Mice were immunized with the recombinant virus.The results showed the recombinant virus could induce HIV 1 specific antibody response.Conclusion:The recombinant fowlpox virus could express the Gag gp120 fusion protein.

Objective:To detect the immune response in mice inoculated by DNA vaccine expressing chimeric gene gag gp120 of HIV 1.Methods:After BALB/C mice were immunized by the eukaryotic expression plasmid pVAXGE,the numbers of T lymphocyte subgroups of spleen,the killing rate of specific CTL from the immunized mice and the titers of serum antibodies during the different time were observed.Results:The lymphocytes of the immunized group by recombinant plasmid pVAXGE proliferated,the specific CTL killing rate of spleen was higher significantly than those of the control group( P

Objective: To adjust the distance between SD sequence and ATG in the same expressive plasmid pLX1 to enhance expression of heterologous bFGF gene in E. coli. Methods: Adjusting the different distance between SD sequence and ATG by Klenow and Mung Bean Nuclease. SDS PAGE and Western blot showed the expressed protein bFGF in E.coli. bFGF proteins were purified by HPHIC, HPGFC and HAC. Biological activity was examined by MTT. Results: Recombinant plasmids pLX2, pLX3 were obtained and the expressive levels were 8.03%, 9.9% respectively. Also the purified bFGF was obtained by HPHIC, HPGFC, HAC and its ED 50 was 2.29 ng/ml. Conclusion: Increasing the bFGF gene dosage by adjusting the distance between SD sequence and ATG could increase the expression level of a desired protein.

Objective: To investigate the curative effects of apoptin on the multidrug-resistant model, of human osteosarcoma cell line (R-OS-732). Methods: 363 bpVP 3 gene was cloned into the vector pIRES1 and the recombinant eukaryon expression vector pIRVP3 was transfected into R-OS-732 cells with liposome. The expression of apoptin gene at transcription level was proved by RT-PCR. The pathological changes of the cells was observed by light microscope and electronmicroscope. The transfected R-OS-732 cells were collected after 48 hours. The genomic DNA extracted from the cells was observed by agarose gel electrophoresis. The apoptosis rate of the cells was analysed by flow cytometry. Results: The expression of apoptin gene at transcription level had been proved by RT-PCR. The construction changes of the two groups were obviously different under light microscope: most of R-OS-732 cells in the transfected group existed in form of being away from the bottom of the culture dish after 24 hours, in form of apoptosis after 48 hours and in form of necrosis after 72 hours; but those cells in the controlled group grow luxuriantly. And under electronmicroscope there was much change of cell nucleus between the two groups. DNA ladder of the transfected cells was observed through agarose gel electrophoresis only one band was observed in the controlled group; but bands of fragmented DNA observed in ositive control group. Flow cytometry analysis showed that the apoptosis rate of the cells in the transfected group was relatively higher than that of the controlls( P

BACKGROUND: Many operations for isolating, purifying and identifying bone marrow mesenchymal stem calls (BMSCs) are complicated and cost much. Also they have great effect on cell activity. Whether whole bone marrow adherent culture can avoid above-mentioned disadvantages remains unclear. At present, many studies huve been done to confirm an effective and low cost method for isolating, purifying and identifying such cells.OBJECTIVE: This study is to in vitro induce and differentiate rat BMSCs by whole bone marrow adherent culture,and to identify the cells.DESIGN: A controlled observational experiment.SETTING: Qingdao University Medical College.MATERIALS: This study was carried out in the Laboratory of Oral Cavity and Laboratory of Molecular Biology (provincial level) Qingdao University Medical College between November 2005 and March 2007. Twenty Wistar rats of either gender, aged 3 to 4 weeks, of SPF grade, weighing 120-150 g, were provided by the Qingdao Laboratory Center. The protocol was carried out in accordance with animal ethics guidelines for the use and care of animals. Fetal bovine serum (FBS, Hangzhou Sijiqing Bioengineering Material Research Institute), alkaline phosphatase (ALP) kit (Nanjing Jiancheng Bioengineering Research Institute), reverse transcription kit (American Promega Corporation) and primer (Shanghai Bioengineering Co.,Ltd.) were used in this study.METHODS: Adult rat BMSCs were isolated and cultured by whole bone marrow adherent culture. They were digested with 2.5 g/L trypse and inoculated at a density of 5 ×107 L-1 in 6-well culture plate. Then, the cells were divided into experimental group and control group. Inducing culture medium was added to experimental group, and the same amount of basic culture medium was added to control group. ① Cell differentiation and calcium tuberculation were observed under the inverted microscope. ② Biological characteristics of induced cells were detected by calcium tubercle Von Kossa and alizarin Bordeaux. ③ALP activity was detected by diazo salt staining. ④Human core binding factor alpha subunit-1 (Cbf α-1), osteocalcin (OCN) and osteoblast-specific Osterix (OSX) mRNA expressions were detected by reverse transcription-polymerase chain reaction (RT-PCR).MAIN OUTCOME MEASURES: ① Induction and differentiation results of cells. ② Biological characteristics of cells induced by rat BMSCs. ③ ALP activity. ④ Cbf α-1, OCN and OSX expressions.RESULTS: ①Inducing culture medium was added in the serial subcultivation. About 9 days later, cell clones were connected to each other. On about 21 to 28 days, some pykno-round mineralized tubercles appeared. Meanwhile,control cells were connected to each other, but they did not form the tubercle. ② In the experimental group, when MSCs were induced for 21 to 28 days, obvious round or oval calcified tubercles were seen by naked eyes. The results of Von Kossa staining exhibited black sediments, and those of alizarin Bordeaux staining exhibited salmon tubercles. Calcium tubercles were not found in the control group. ③The ALP activity after 2 weeks of induction was obviously increased in the experimental group, but was relatively weak in the control group. ④In the experimental group,Cbf α-1, OCN and OSX expressions were significantly increased after induction.CONCLUSION: After being in vitro induced and differentiated by whole bone marrow adherent culture, rat BMSCs exhibited morphological and biological characteristics similar to typical osteoblasts.