Objective:Osteoprotegerin (OPG) is a key molecule negatively regulating osteoclast differentiation and activation; and the conserved mycobacterial heat shock 70 (HSP70) peptide p111-125 has also been found to inhibit inflammation reactions in chronic arthritis. BaculoDirectTM baculovirus expression system was selected to express recombinant OPG-HSP70 in insect cells.Methods:The human functional fragment (p22-194) of OPG and functional fragment (p111-125) of mycobacterial HSP70 gene were cloned into the transfer vector pENTRTM/SD/D-TOPO. The recombinant plasmid was performed an LR reaction with the BaculoDirectTM Linear DNA to generate recombinant baculovirus DNA. The cultured Sf9 insect cells were directly transferred with the recombinant baculovirus DNA,and the pure recombinant baculovirus was obtained. Then recombinant baculovirus was infected Sf9 insect cells again to express the OPG-HSP70 gene.Results:The target protein was detected at the time of 48h post infection,reached at highest yield at the time of 72h post-infection. A 28kDa protein immunostaining band was detected by Western blotting from lysate of those cells.And the purified protein was obtained by using Ni-NTA system. Functional stuies on the fusion protein showed it significantly reduce osteoclast cell number[(3.10?0.640) cells under each microscope field in treatment group by comparing to (10.70?0.817)cells in the control group] in the osteoclast inhibition test,and reduce the inflammation reaction in a delayed type hypersensitivity (DTH) mice model (P

Objective To study the relationship between environmental chemical elements,vires infection and dilated cardiomyopathy(DCM).Methods In 2008,233 patients with DCM(case group)and 150 patient with stable angina(control group)were chosen in Liaocheng People's Hospital and Yanggu People's Hospital,Shandong province.Population distribution and disease history were surveyed in the two groups.Human myocardial antibody IgG(AMA-IgG),Coxsackie B virus IgG(CBV-IgG),Adenofirus antibody IgG(ADV-IgG)were detected by ELISA in both the case group and the control group.Serum trace elements were detected in the two groups.The general chemical and toxicological indicators in drinking water of the high-and the low-incidence aireas of the disease were control group[60.00%(90/150),χ2=13.80,P<0.01)].Per capita annual income(Yuan,RMB)in the case group (3207.82±618.51)was lower than that of the control group[(5086.61±886.12),t=24.40,P<0.01].Personal alcohol consumption in the case group[(175.00±160.50)g/d]was higher than that of the control group[(110.22±100.03)g/d,t=4.40,P<0.01)].The rate of myocarditis in the case group[5.15%(12/233)]was higher than ADV-IgG in the cage group were 7.78%(7/90),6.67%(6/90)and 6.67%(6/90),respectively.Compared with those in the control group[3.33%(2/60),5.00%(3/60)and 5.00%(3/60),χ2=1.26,0.18,0.18,all P>0.05],no mg/L]in drinking water of the high-incidence areas were significantly higher than that of iron[(0.39±0.67)mg/L,t=2.11,P<0.05]and that of manganese[(0.15±0.14)mg/L,t=3.01,P<0.01]in the low-incidence arefas.The content of semm iron[(69.1±57.8)μmol/L]in the case group evidently exceeded the normal range(15.6-35.9 μmol/L)and obviously higher than that in the control group[(20.0±17.5)μmol/L,t=5.04,P<0.01].Conclusions Theso data do not support that DCM is related with persistent virus infection and autoimmunization.DCM is probably related with low incomes,high alcohol consumption,myocarditis,high iron and manganese contents in drinking water and high content of serum iron.

Objective To explore the tumor cell apoptosis induced by the effect in vitro of eukaryotic ex pression of full-length TRAIL cDNA and its extracellular region. Methods The eukaryotic expression vectors for both forms of the cDNA acquired from the fetal heart cDNA library were constructed. After gene transfecting, the stable expression cell lines were obtained by G418 screening. Results The supernatants from tansfectants could induce apoptosis of different tumor cell lines in vitro, and an enhanced effect was observed by adding TFAR19 (TF-1 cell apoptosis-related protein 19), a novel apoptosis gene product discovered in our laboratory. Conclusion Eukaryotic expression products of TRAIL can induce apoptosis of the tumor cells, and TFAR19 could enhance the effect on apoptosis of tumor cells.

Using the weight-average molecular weight 50 000 polylactic acid (PLA) as a carrier, and a certain proportion of erythromycin (EM) and prednisone acetate (PNA) to mixed prepare the compound erythromycin sustained release preparation (sustained-release tablets). Using ultraviolet spectrophotometry and high performance liquid chromatography (HPLC) to detect separately the release amount of EM and PNA in vitro medium. The sustained-release tablets release for about 21 days, the average content of EM is 99.7 mg/table, RSD = 0.82%; and the average content of PNA is 10.03 mg/table, RSD = 0.93%. Within 21 days, the cumulative releases of EM and PNA are 86.1% and 78.3%, respectively. The drug release is steady and slow after 5 days, the burst release phenomenon in early stage is more significant. The results showed that the sustained-release tablet preparation method is feasible, the release performance is good and the clinical efficacy is significant.
Chromatography, High Pressure Liquid ; Delayed-Action Preparations ; administration & dosage ; chemistry ; therapeutic use ; Drug Carriers ; Drug Combinations ; Erythromycin ; administration & dosage ; chemistry ; therapeutic use ; Humans ; Lactic Acid ; administration & dosage ; Polyesters ; Polymers ; administration & dosage ; Prednisone ; administration & dosage ; chemistry ; therapeutic use ; Sinusitis ; drug therapy ; Spectrophotometry, Ultraviolet ; Tablets

OBJECTIVETo investigate the effect of the arthroscopic procedure on the patients with recurrent anterior shoulder dislocation.

METHODSFrom January 2001 to March 2006,52 patients with recurrent anterior shoulder dislocation were treated by arthroscopy. Among them 44 patients were followed up for 12 to 54 months(on average of 26 months). The data of the 44 patients was reviewed. Three evaluation systems, University of California at Los Angeles Shoulder Scores (UCLA), Simple Shoulder Test (SST) and Dawson, were used. The study evaluated the effect based on the Dawson system by the factors as: age, course of the disease, frequency of dislocation and relocation methods and the range of shoulder movement.

RESULTSThe ratio of recurrent dislocation after operation was 4.5%. Assessing through 3 evaluation systems, UCLA, SST and Dawson, results were similar: the follow-up evaluation were extraordinarily different from preoperative assessment, and the rating of good or excellent at the time of the final follow-up reached 91% higher. Based on the Dawson system, the evaluation results had no statistic diversity according to such factors as: age, course of the disease, frequency of dislocation and relocation methods.

CONCLUSIONSThe effects of arthroscopic surgery to the recurrent anterior shoulder dislocation are satisfied and evidence-proved. It is a good option for both common patients and athletes.

Adolescent ; Adult ; Arthroscopy ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Radiography ; Recurrence ; Retrospective Studies ; Shoulder Dislocation ; diagnostic imaging ; surgery ; Treatment Outcome

To investigate the adjuvant effect of plasmid DNA encoding superantigen SEA (D227A) (pmSEA) on immune responses induced by HBV DNA vaccine containing HBV preS2 and S antigen in BABL/c (H-2d). BALB/c mice were immunized intramuscular injection with HBV DNA vaccine (pHBVS2S) mixed with or without pmSEA plasmid. Antibodies againat HBV PreS2 and S antigen in the sera were accessed by Anti-HBs ELISA, and the HBsAg specific cytotoxic T lymphocytes (CTLs) activity was determined by 5 Chromium Release Assay. The HBs peptide-specific IFN-gamma secreting T cells were detected by ELISPOT. Anti-HBs antibody titers and CTLs activity in mice immunized with pmSEA + pHBVS2S group were significant higher (P < 0.05) than pHBVS2S DNA vaccine group. The ratio of IgG1/IgG2a (0.282) was apparently different from the group immunized with peptide (10). Mice immunized with HBV DNA vaccine plus adjuvant produce higher titer of IgG1 and IgG2a antibodies against HBV S antigen 1.36 and 1.73 time higher than that without adjuvant respectively. HBs peptide--specific IFN-gamma secreting T cells increased 2 - 3 times by the pmSEA adjuvant, compared to DNA vaccine group. HBV DNA vaccine (pHBVS2S) induces humoral and cellular immuno-responses in BALB/c mice, and the responses could be significantly boasted by the plasmid encoding mSEA. Therefore the pmSEA was a potential adjuvant for DNA vaccines.
Adjuvants, Immunologic ; Animals ; Enterotoxins ; immunology ; Hepatitis B ; immunology ; prevention & control ; therapy ; Hepatitis B Antibodies ; blood ; Hepatitis B Vaccines ; immunology ; Interferon-gamma ; secretion ; Mice ; Mice, Inbred BALB C ; Staphylococcus aureus ; immunology ; Superantigens ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccination ; Vaccines, DNA ; immunology

This study was aimed to investigate the sensitizing effect of recombinant human PDCD5 (rhPDCD5) protein on chemotherapy of U937 cell line and its mechanism. The flow cytometry was performed to assess the changes of cell apoptosis and cell cycle influenced by rhPDCD5. Hochst 33258 staining was used to observe morphology of the apoptotic cells. The activity change of caspase-3 was detected to analyse the possible mechanisms of rhPDCD5-induced apoptosis. RT-PCR was performed to observe the expression level of drug-resistant genes. The results showed that the percentage of apoptotic cells and the activity of caspase-3 remarkably increased in U937 cells treated with rhPDCD5 combined with chemotherapeutic drug; the cell cycle arrest induced by anti-tumor drug was also enhanced when combined with rhPDCD5; meanwhile, the expression levels of drug-resistant genes were down-regulated in jointly treated U937 cells. It is concluded that the chemosensitizing mechanisms of rhPDCD5 are complex. rhPDCD5 may increase the cytotoxicity of anti-tumor drugs by promoting the caspase-3-related apoptosis, influencing cell cycle, decreasing the expression of drug-resistant genes and reversing drug-resistance.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; pharmacology ; Caspase 3 ; metabolism ; Cell Cycle ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans ; Neoplasm Proteins ; pharmacology ; Recombinant Proteins ; pharmacology ; U937 Cells

·AIM: To investigate the relationship between the thickness of choroid and the development of idiopathic macular epiretinal membrane (IMEM). ·METHODS: A retrospective analysis on 48 cases (48 eyes) of idiopathic macular epiretinal membrane patients (the IMEM group) was taken in our hospital from January 2014 to December 2016, and 50 right eyes in 50 healthy persons with physical examination were selected the control group, comparison on subfoveal choroidal thickness (SFCT) levels of sicked eyes, normal eyes in IMEM group and the control group were made, postoperative SFCT level change of sicked eyes in IMEM patients and normal eyes were investigated via follow-up, and analysis on correlation between postoperative choroidal thickness and the best corrected visual acuity was taken. ·RESULTS: The SFCT of sicked eyes in IMEM group 362.22±40.75μm was significantly lower than that of the contralateral eyes (410.56 ± 38.45μ m) and the right eyes of the control group (420.73±39.63μ m), and data of the contralateral eyes was lower than right eyes of the control group, distinct difference was shown between groups(P<0.05). The IMEM group's SFCT of sicked eyes and normal eyes at postoperative 1wk had no significant difference with that before operation (P>0.05), and at postoperative 1mo,SFCT of sicked eyes and normal eyes evidently increased, showing sharp difference compared with that before operation (P<0. 05). After that, the SFCT value stabilized, but there was no obvious difference between the sicked eyes and healthy eyes at postoperatively 1mo (P>0.05). The numbers of patients whose postoperative BCVA ≥ 0. 5 with different preoperative SFCT values had statistically significant differences (P<0.05), and those with BCVA ≥0.5 of SFCT values> 380μ m were significantly higher than those with <320μ m and 320μ m-380μ m groups. Fisher exact probability analysis showed that the differences were significant. Pearson analysis showed that there was a positive correlation between the postoperative choroid thickness and the best corrected visual acuity of IMEM group (r=0.629,P<0.05). · CONCLUSION: Choroidal thinning may be an important cause of IMEM, and preoperative choroidal thickness also has an influential effect on postoperative visual recovery.

To investigate the activity of CKLF1 on the proliferation and differentiation of bone marrow cells. Methods Human low density bone marrow cells and mouse bone marrow cells were plated in 96-well microplate and supernatants from transfected COS-7 cell culture were added. The cell proliferation was assayed by MTT method after 5 days incubation. The enhancing effect of CKLF1 on the colony forma tion of human hematopoietic progenitor cells was identified in semi-solid culture. Results CKLF1 has obvi ous enhancing effect on both human and mouse bone marrow cells, it can stimulate the colony formation of human hematopoietic stem cells and has synergistic action with GM-CSF. Conclusion CKLF1 can pro mote the proliferation and differentiation of bone marrow cells.

OBJECTIVETo investigate the status of lymph node metastases (LNM) of esophageal carcinoma and to identify the risk factors.

METHODSClinical data of 308 patients who underwent esophagectomy with three-field lymphadenectomy during January 2006 and December 2010 were reviewed. Characteristics of LNM were studied.

RESULTSThe average number of dissected lymph nodes was 35.6 ± 14.5 in 308 patients. There were 197 patients(64%) had LNM. Logistic regression analysis showed that lymphatic vessel invasion(P=0.019) and deep tumor invasion(P<0.001) were risk factors of LNM. The highest LNM site was paratracheal node(25.0%). The incidence of cervical LNM was 14.1% in the middle thoracic carcinoma, higher than that of upper thoracic (7.3%) and lower thoracic (8.3%). Rate of LNM was lower in upper thoracic carcinomas than that in middle or lower ones(P=0.001). No significant difference of LNM was found among upper, middle and lower thoracic carcinoma for cervical or thoracic nodes. Lymphatic vessel invasion(P<0.001) and metastases in paratracheal lymph nodes (P=0.014) were risk factors for cervical LNM.

CONCLUSIONSLNM of esophageal carcinoma can be found in both directions vertically and skipped metastasis. Paratracheal lymph nodes involvement is an indicator for cervical lymphadenectomy in thoracic esophageal carcinoma.

Aged ; Carcinoma, Squamous Cell ; pathology ; Esophageal Neoplasms ; pathology ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Lymphatic Vessels ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Risk Factors