To enhance the immunogenicity of DNA and adenoviral vector vaccines expressing HIV-1 subtype B gp160, human interleukin 15 (hIL15) DNA adjuvant (pVR-hIL15) was constructed. BALB/c mice received DNA prime/protein boost immunization with pVR-HIVgp160/Ad5-HIVgp160 alone or combined with pVR-hIL15. Cellular and humoral immune responses were evaluated by IFN-gamma enzyme-linked immunosorbent spot assay and enzyme-linked immunosorbent assay, respectively. Compared with those immunized with vaccines alone, the mice immunized with vaccines combined with pVR-hIL15 had significantly increased specific cellular response and antibody titer (P < 0.05). It suggests that the IL15 DNA adjuvant can enhance the immune responses induced by prime-boost regimen using DNA and adenoviral vector encoding HIV-1 subtype B gp160.
Adenoviridae ; genetics ; Adjuvants, Immunologic ; Animals ; Antibodies, Viral ; immunology ; Antibody Specificity ; Female ; Genetic Vectors ; genetics ; HIV Envelope Protein gp120 ; immunology ; HIV Envelope Protein gp160 ; genetics ; immunology ; HIV Envelope Protein gp41 ; immunology ; Humans ; Immunity, Cellular ; Immunity, Humoral ; Interleukin-15 ; genetics ; Mice ; Mice, Inbred BALB C ; Vaccines, DNA ; genetics ; immunology

OBJECTIVETo study the stereoselectivity of R-(+) and S-(-)-propranolol glucuronidation and metabolic interaction between R(+)- and S-(-)-propranolol.

METHODSA RP-HPLC analytical method was developed for determination of R-(+)-and S-(-)-propranolol glucuronide (PG) incubated with rat hepatic microsome induced with phenobarbital (PB). The method was applied to investigate the stereoselectivity metabolism of racemic propranolol glucuronidation in vitro.

RESULTIn control and PB group, the concentration of R-(+)-PG produced at different substrates was higher than that of S-(-)-PG. Compared with the control, the V(max) and Cl(int) for R(+)-and S-(-)-propranolol increased significantly the K(m) for R(+)-propranolol was elevated, while that for S-(-) propranolol was decreased.

CONCLUSIONThere is a stereoselectivity in glucuronidation of propranolol in rat hepatic microsome induced with PB and R-(+)-propranolol is preferred. Metabolic interaction between R-(+)-and S-(-)-propranolol exists with a concentration-dependent mode.

Animals ; Enzyme Induction ; Microsomes, Liver ; enzymology ; Phenobarbital ; pharmacology ; Propranolol ; analogs & derivatives ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stereoisomerism

Objective To investigate the clinical significance of serum cytokines concentrations and A- PACHE scores in evaluating the illness state for critical trauma patients.Methods A clinical prospective self-control trial was performed,in which 36 patients admitted to ICU by SIRS were enrolled.Objects were divided into mild and severe trauma group according to APACHE score.The TNF-?and IL-6 concentrations were determined on the 1st, 3rd and 5th day of admission,the APACHE score were assessed at the same time.Statistic analysis was performed according to this group.Results The TNF-?concentrations decreased continuously in the following days while IL-6 decreased from the 7th day in the mild trauma group.In the severe trauma group the TNF-?and IL-6,APACHE score concentrations kept increasing.There was a significant difference of TNF-?and IL-6 concentrations between severe trauma and mild trauma group.Conclusion Dynamic measurement of TNF-?and IL-6 concentrations with APACHE score provide great help to evaluate the illness state and predict the prognosis.

Objective To determine kinetics of TNF-α and miR-146a (microRNA-146a)expressions in lipopolysaccharide (LPS)-induced NR8383 alveolar macrophages (AM) at different intervals and their relationships in order to explore regulatory effect and mechanism of miR-146a on alveolar macrophages inflammatory responses.Methods NR8383 alveolar macrophages were seeded in a 6-well plate,and stimulated with 1 μg/ml of LPS for 0 h,3 h,6 h and 12 h separately after 90 min.Cells were harvested and supernatant were collected 0 h,3 h,6 h and 12 h after incubation.The expressions of miR146a and TNF-α mRNA in cells were detected by real-time qPCR and the levels of TNF-α protein in the supematant of cells were assayed by enzyme-linked immunosorbent assay ( ELISA ).Pearson correlation analysis was used to analyze the correlation between miR-146a and TNF-α mRNA.Results ①The level of TNF-α protein in the supernatant of cell was significantly increased 3 h after LPS challenge ( 359.80 ±57.54) pg/ml (P ＜0.01 ),and peaked 12 h later (729.22 ±50.40) pg/ml (P＜0.01 ) ; ②the expression of TNF-α mRNA peaked 3 h after LPS challenge (67.48 ±24.52) fold,P ＜0.01 ),and then decreased gradually; ③the expression of miR-146a mRNA increased continuously until 6 h or 12 h after LPS challenge 6 h:(5.33 ±0.81) fold,12 h:(8.21 ±1.19) fold,(P＜0.01),and it showed an upward tendency;④ the expression of miR-146a mRNA was negatively correlated with TNF-α mRNA ( r =-0.895,P ＜0.01).Conclusions The miR-146a mRNA showed a negative correlation with TNF-α mRNA present in lipopolysaccharide-stimulated alveolar macrophages,suggesting miR-146a mRNA involved in regulating the inflammatory response of alveolar macrophages.

OBJECTIVETo study the osseointegration of the nanophase hydroxyapatite biograde-coated implants and host bone.

METHODSNanophase hydroxyapatite biograde-coated implants, hydroxyapatite biograde-coated implants and noncoated Ti-6Al-4V implants were inserted into both femoral of Beagle canines the tissue response, dynamic osteogensis and SEM were studied at 4, 8 and 12 weeks.

RESULTSThe degradation of nanophase hydroxyapatite was slower than hydroxyapatite, dynamic osteogensis and the form of osteoblast were better than the control groups.

CONCLUSIONThe biological reaction of Nanophase hydroxyapatite biograde-coated implants is better than hydroxyapatite coated implant.

Animals ; Bone Substitutes ; chemistry ; Coated Materials, Biocompatible ; chemistry ; Dogs ; Durapatite ; chemistry ; Male ; Materials Testing ; Nanoparticles ; Osseointegration ; physiology ; Surface Properties

Objective To analyze and compare the MRI appearances of ovarian thecoma with pathologic findings in order to improve the knowledge of the disease.Methods Nineteen cases of ovarian thecoma confirmed by histopathology were analyzed.MRI morphological characteristics and signal intensity of the lesions were observed and compared with findings of pathomorphology.Correlation analysis between tumor size and amount of ascites was made.Results Ovarian thecoma displayed iso-or hypointense signal on T_1WI and significant hypointensity in the focal lesion on T_2WI.Hyperintensity occurred when cystic degeneration of the lesions existed.Fibrous septation was detected in some lesions.After enhancement,most lesions showed mild early enhancement with slight increase on the delayed phase.Pathological necrosis and cystic degeneration were seen in 9 cases which corresponded to the number and shape of the cystic lesions on MRI.A large amount of collagen hyperplastic was found between the oncocytes microscopically in 15 cases, which displayed significant hypointensity in the focal lesion on T_2WI;another 4 cases showed relatively less amounts of collagen hyperplastic and more oncocytes,which appeared moderate intensity in the focal leisom on T_2WI.The amount of ascites was not significantly correlated with the lesion size(r=0.43,P=0.10). Conclusions Hypointensity on T_2WI and mild enhancement pattern due to poor blood supply are the characteristics of ovarian thecoma.The MR findings can reflect the pathologic features of the tumor,which is helpful in the diagnosis and differential diagnosis.

Objective To analyze the normal anatomy of petrosal vein and the space adjacent relationship between the petrosal vein and the homolateral trigeminal nerve by balanced turbo field echo (B-TFE) Cine-MR imaging and enhanced T1 high-resolution isotropic volume excitation (e-THRIVE)imaging.Methods Forty-one patients with facial spasm and epileptiform neuralgia were selected and taken a scan with ACHIEVA NOVA DUAL A-serial 1.5T MR machine using the BTFE and e-THRIVE series.The space adjacent relationship between the petrosal vein and the trigeminal nerve in cerebellopontine angle were observed.Results The 82 sides of petrosal veins and homolateral trigeminal nerves (41 cases) were displayed well.Petrosal veins were located in cavitas subarachnoidealis,partly in free state; the number of trunk ofpetrosal vein could be 1,2,3,respectively,responding to 70 (86%),10 (12%),2 (2%).The petrosal veins located in the dorsal-lateral of trigeminal nerve were found in 74 sides (37 cases,91%,by BTFE,e-THRIVE series),the petrosal veins located in the ventri-lateral of trigeminal nerve in 6 sides (3 cases,7%,by e-THRIVE series),and the petrosal veins located in the right upon trigeminal nerve in 2 sides (1 cases,2%,by BTFE series).Conclusion B-TFE MR imaging and e-THR/VE imaging,showing the petrosal veins and trigeminal nerves clearly and evaluating their relationship accurately,can provide information of topographic anatomy before microsurgical vascular decompression.

Objective To explore the applied value of balanced turbo field echo (BTFE)Cine-MR imaging and enhanced T1 high-resolution isotropic volume excitation (e-THRIVE) imaging in patients with trigeminal neuralgia (TN) and hemifacial spasm (HFS) before performing microvascular decompression (MVD).Methods One hundred and twenty-one patients (74 patients with TN and 47 with HFS),admitted to our hospital from April 2009 to December 2011,were chosen in our study; a scan with ACHIEVA NOVA DUAL A-serial 1.5 T MR Machine using the BTFE and e-THRIVE serial besides conventional head MR imaging was performed on these patients.All the MR imaging data were retrospectively analyzed.Results All the patients were confirmed by microvascular decompression.The facial nerves of 74 patients with TN and 47 with HFS were displayed well.Responsible capillaries were found in 118 patients (98％):the related vessel was superior cerebellar artery in 52 patients (70％) of 74 patients with TN and anterior inferior cerebellar artery in 29 patients (625) of 47 patients with HFS.The compression style included oblique cross,parallel or vertical compression.Conclusion BTFE MR imaging and e-THRIVE imaging can show the trigeminal nerve,facial nerve and microvascular vessel in the posterior cranial fossa clearly; combining space-link method and multiplanar reconstruction,they can help us find the related vessel and provide preoperative guidance before microvascular decompression for patients with TN and HFS.

BACKGROUNDSignal transducer and activator of transcription 3 (STAT3) is usually constitutively activated in a variety of malignancies. It directly contributes to tumorigenesis, invasion, and metastasis. The surgical treatment of breast cancer has made no breakthroughs in terms of treatment effect, in spite of its long history. Current biotherapies bring a note of optimism to breast cancer treatment. To explore the possibility of a siRNA targeted STAT3 blocking treatment for over-activated tumor cells, we evaluated the efficacy of a STAT3 siRNA on human breast cancer cells in vitro and in vivo.

METHODSThree MCF-7 human breast cancer cell lines were tested: control MCF-7 cells, non-specific siRNA transfected MCF-7 cells and STAT3 siRNA transfected MCF-7 cells. Expression of STAT3 in MCF-7 cells was inhibited by RNA interference (RNAi). The STAT3 mRNA and protein levels were detected by semi-quantity RT-PCR and Western blotting. Cell proliferation and apoptosis were determined by MTT method and flow cytometry. The three groups of MCF-7 cells mentioned above were transplanted subcutanuously into nude mice and their tumorgenic ability observed. The STAT3 mRNA and protein levels of the samples from tumors in different groups were determined by semi-quantity RT-PCR and Western blotting and compared.

RESULTSIn STAT3 siRNA transfected MCF-7 cells, the expressions (STAT3/β-actin) of STAT3 mRNA (0.327 ± 0.020) and protein (0.153 ± 0.006) were significantly lower than that in control MCF-7 cells (mRNA 1.093 ± 0.018, protein 1.374 ± 0.022) and non-specific siRNA transfected MCF-7 cells (mRNA 1.035 ± 0.050, protein 1.320 ± 0.033) (P < 0.05). MTT showed that cell proliferation was significantly reduced and the cell growth inhibition ratio in the STAT3-siRNA group was (44.00 ± 5.10)%, significantly higher than that in non-specific siRNA transfected MCF-7 cells ((16.10 ± 1.05)%, P < 0.05). Flow cytometry results showed that more apoptosis was observed in the STAT3-siRNA group. The rate of apoptosis was (14.79 ± 0.22)%, much higher than in control MCF-7 cells (7.06 ± 0.71) and non-specific siRNA transfected MCF-7 cells (8.45 ± 0.43) (P < 0.05). The tumor growth in the STAT3 siRNA transfected MCF-7 cells was significantly slower than in the two control groups. On the 22th day after transplantation the tumor weight ((21.40 ± 10.57) mg) and volume ((41.15 ± 12.17) mm(3)) in the STAT3 siRNA transfected group were significantly lower than in control group (weight (88.60 ± 12.16) mg, volume (118.45 ± 24.68) mm(3)) and non-specific siRNA transfected group (weight (57.20 ± 21.86) mg, volume (101.36 ± 21.90) mm(3)) (P < 0.05). Both the STAT3 mRNA and protein levels in the tumors from the STAT3 siRNA transfected group were significantly lower than in the tumors from the two control groups.

CONCLUSIONSSTAT3 siRNA can effectively silence the STAT3 gene in vitro and in vivo, increase cell apoptosis rate and significantly decrease cell proliferation, which inhibits the growth of breast cancer cell in vitro. Tumor growth of xenograft mice is significantly inhibited. The results obtained in vivo are in consistency with those in vitro. STAT3 may be a novel therapeutic target for breast cancer and RNA interference has potential clinical application.

Animals ; Apoptosis ; Cell Line, Tumor ; Female ; Humans ; Mammary Neoplasms, Experimental ; pathology ; therapy ; Mice ; Mice, Nude ; RNA, Small Interfering ; genetics ; STAT3 Transcription Factor ; antagonists & inhibitors ; genetics ; Xenograft Model Antitumor Assays

OBJECTIVETo study the cultural method and identification of human umbilical vein endothelial cells (HUVECs), and investigate the expression of tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains(Tie-2) in HUVECs.

METHODSHUVECs were isolated from umbilical veins by the technique of irrigative digestion, and were cultivated in plates. The cells were identified by VIII monoclonal antibody. Tie-2 mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and SABC immunocytochemistry.

RESULTSHUVECs could adhere to the plates completely after 24 hours, and confluence a monolayer 4-5 days later. The band of Tie-2 mRNA was obviously and the expression of Tie-2 protein was strongly positive by immunocytochemistry in HUVECs. The positive rate was over 85%.

CONCLUSIONHighly purified endothelial cells were isolated. And there were overexpression of Tie-2 in HUVECs.

Cells, Cultured ; EGF Family of Proteins ; Endothelial Cells ; Human Umbilical Vein Endothelial Cells ; Humans ; Immunoglobulins ; TYK2 Kinase ; Umbilical Veins