This study is to prepare the in situ forming sustained-release injection which can perform sustained release behavior at the periodontal site for 7 days and to evaluate its in vitro and in vivo properties. After preparation of in situ forming sustained-release injection the in situ time was studied. And the surface of the solid injection was characterized by SEM. The rheological curve at 0 degrees C, 25 degrees C, 37 degrees C was determined and the impact of the temperature on the viscosity was examined. The in vitro release behavior was investigated. At last, rabbit periodontitis model was established to study its pharmacokinetics. The injection was stable, hard to stratify and decompose. The in situ forming time was about 6 seconds. It can easily adhere into periodontal pockets. There were lots of holes on the surface of the solid injection for the drug to diffuse. The drug releasing curves could be fit by Korsmeyer-Peppas equation. The drug smoothly released for 7 days at pH 7.4 PBS buffer with a very slight burst release and maintained a certain concentration. In vivo pharmacokinetics results indicated that after administration with the in situ forming injection, achievement of tinidazole (TNZ) concentration in gingival crevicular fluid (GCF) was more comparable and long-lasting than usual solution of TNZ management and relatively constant TNZ levels were attained until 168 h. All these results supported the prospect of tinidazole in situ forming sustained-release injection in clinical applications.

China ; Genes, Bacterial ; Sequence Analysis ; Streptococcus suis ; classification ; genetics ; isolation & purification ; Virulence Factors ; genetics

OBJECTIVETo study the specific cellular immune response induced by a recombinant HBV DNA vaccine and evaluate the potential therapeutic effect of this vaccine against HBV.

METHODSA series of HBV epitopes were screened and the optimal combinations of these gene fragments identified to construct the target gene HS which incorporated appropriate flanking sequences. HS was inserted in pVAX1 to construct the recombinant plasmid PVAX-HS. Generation of specific cytotoxic T lymphocytes (CTLs) induced by PVAX-HS in HLA-A2 mice was observed by 51Cr assay and the activity of the CTLs evaluated using ELISPOT assay.

RESULTSPVAX-HS was constructed successfully, which induced specific CTLs with strong activities after immunization in mice as compared with the control group that yielded negative results.

CONCLUSIONPVAX-HS DNA immunization can induced specific cellular immune response in mice, suggesting the potential therapeutic effect of this HBV vaccine.

Animals ; Hepatitis B Vaccines ; genetics ; immunology ; Hepatitis B virus ; genetics ; immunology ; Humans ; Immunization ; methods ; Interferon-gamma ; biosynthesis ; Mice ; Recombinant Fusion Proteins ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; metabolism ; Vaccines, DNA ; genetics ; immunology

This study is to prepare the in situ forming sustained-release injection which can perform sustained release behavior at the periodontal site for 7 days and to evaluate its in vitro and in vivo properties. After preparation of in situ forming sustained-release injection the in situ time was studied. And the surface of the solid injection was characterized by SEM. The rheological curve at 0 degrees C, 25 degrees C, 37 degrees C was determined and the impact of the temperature on the viscosity was examined. The in vitro release behavior was investigated. At last, rabbit periodontitis model was established to study its pharmacokinetics. The injection was stable, hard to stratify and decompose. The in situ forming time was about 6 seconds. It can easily adhere into periodontal pockets. There were lots of holes on the surface of the solid injection for the drug to diffuse. The drug releasing curves could be fit by Korsmeyer-Peppas equation. The drug smoothly released for 7 days at pH 7.4 PBS buffer with a very slight burst release and maintained a certain concentration. In vivo pharmacokinetics results indicated that after administration with the in situ forming injection, achievement of tinidazole (TNZ) concentration in gingival crevicular fluid (GCF) was more comparable and long-lasting than usual solution of TNZ management and relatively constant TNZ levels were attained until 168 h. All these results supported the prospect of tinidazole in situ forming sustained-release injection in clinical applications.
Animals ; Antitrichomonal Agents ; administration & dosage ; pharmacokinetics ; Delayed-Action Preparations ; Drug Carriers ; Drug Compounding ; methods ; Endotoxins ; Gingival Crevicular Fluid ; metabolism ; Injections ; Periodontal Pocket ; metabolism ; Periodontitis ; chemically induced ; metabolism ; Polyesters ; chemical synthesis ; pharmacokinetics ; Polyethylene Glycols ; chemical synthesis ; pharmacokinetics ; Rabbits ; Random Allocation ; Rheology ; Tinidazole ; administration & dosage ; pharmacokinetics

Objective To investigate the clinical treatment and outcomes of severe community-acquired pneumonia (CAP) complicated with mediastinal emphysema after renal transplantation. Methods Clinical data of9 patients with severe CAP complicated with mediastinal emphysema after renal transplantation were retrospectively analyzed. The acute physiology and chronic health evaluationⅡ(APACHEⅡ) and oxygenation index were recorded when the patients were admitted to the intensive care unit (ICU). The complications of mediastinal emphysema and corresponding treatment were observed. The treatment course during the ICU, mortality rate in ICU, ICU stay time and hospital stay time were recorded. All patients underwent pathogenic examinations. Results The APACHEⅡ score of9 patients with severe CAP complicated with mediastinal emphysema after renal transplantation was 14 (8-21) scores and the oxygenation index was 150 (133-189) mmHg. Among 9 patients, 3 cases were infected by bacteria alone, 3 cases were infected by bacterial infection combined with viral infection, 1 case was infected by mycobacterium tuberculosis complicated with other bacterial infection and 1 case was viral infection. No pathogenic evidence was detected in the remaining 1 patient. Mediastinal emphysema complicated with subcutaneous emphysema occurred in 7 cases and pneumothorax occurred in 6 cases. Treatment methods included anti-infection, modified immunosuppressive program, mediastinal drainage, thoracic closed drainage, subcutaneous incision and extracorporeal membrane oxygenation (ECMO) treatment. Six patients received invasive mechanical ventilation (IMV), 2 received non-invasive positive pressure ventilation (NIV) and 1 received high-flow nasal oxygen cannula (HFNC). Among 9 patients, the mortality rate in ICU was 6/9, the remaining 3 patients were recovered and discharged, the ICU stay time was 26 (17-40) d, and the total hospital stay time was 27-61 d. Conclusions Mediastinal emphysema is a serious complication of patients presenting with severe CAP after renal transplantation with a high mortality rate. For these patients, imaging evaluation, timely drainage and full sedation should be strengthened, and ECMO treatment should be delivered when necessary.

OBJECTIVETo investigate the changes in cell proliferation and retinoic acid receptor gamma (RARgamma) mRNA expression in normal human keratinocytes after acitretin treatment and/or narrow-band ultraviolet-B irradiation.

METHODSNormal human keratinocytes were exposed to irradiation with 100 mJ/cm square NB-UVB and/or subsequent 12-hour incubation with 1x10(-6) mol/L acitretin, and the expression of RARgamma mRNA in the cells was examined using RT-PCR and real-time quantitative RT-PCR.

RESULTSA 0.9- and a 2.3-fold increase in RARgamma mRNA expression was induced in the cells by exposure to 100 mJ/cm square NB-UVB and 10(-6) mol/L acitretin, respectively, and the expression was synergistically enhanced by 2.8-fold after their combined treatment.

CONCLUSIONUpregulated expression of RARgamma mRNA can be associated with keratinocyte growth inhibition after treatment with acitretin and NB-UVB irradiation.

Acitretin ; pharmacology ; Cells, Cultured ; Humans ; Keratinocytes ; drug effects ; radiation effects ; RNA, Messenger ; metabolism ; Receptors, Retinoic Acid ; metabolism ; Ultraviolet Rays

OBJECTIVETo investigate the role of tazarotene induced gene-2 (TIG2) in psoriasis vulgaris.

METHODSTIG2 protein and mRNA expressions in normal tissues, psoriatic lesions and uninvolved skin tissues were detected by immunohistochemistry and in situ hybridization, respectively.

RESULTSTIG2 protein and mRNA were expressed in all the layers of normal and uninvolved epidermis. TIG2 expression was detected in the upper layers of the stratum spinosum of the marginal region of the psoriatic lesions, but not in the central area of the lesions. TIG2 expression was significantly lower in the basal layers of the central area of the paoriasis than that in the normal skin and uninvolved tissues (P < 0.01), and also lower in the marginal regions of the lesions (P < 0.01).The suprabasal layers of the marginal region in the lesion showed significantly lower TIG2 expression than the central area of the lesion (P < 0.01).

CONCLUSIONTIG2 may maintain the normal differentiation of epidermal keratinocytes and implicate in the pathogenesis and development of psoriasis vulgaris.

Adolescent ; Adult ; Chemokines ; Chemotactic Factors ; biosynthesis ; genetics ; Female ; Humans ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; genetics ; Keratinocytes ; metabolism ; Male ; Middle Aged ; Psoriasis ; genetics ; metabolism ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVETo investigate the feasibility of obtaining of a highly pure protein of human endothelial overexpressed lipopolysaccharide-associated factor 1 (EOLA1) with metal chelation chromatography.

METHODSInclusion bodies of the E. coli transformed with EOLA1 gene were extracted and washed with BugBuster Protein Extraction Reagent. The primary purified products were purified by His. Bind Resin Chromatography under denaturing condition and dialyzed for renaturation, and then were analyzed with SDS-PAGE, Western blotting and peptide mass fingerprinting (PMF).

RESULTSEOLA1 was mainly expressed in E. coli as insoluble inclusion bodies. The protein content in the primary extracted inclusion bodies accounted for over 75%, and it accounted for more than 90% after chromatography and renaturation. It was indicated by PMF that the targeted protein peptide overlaid many of designed protein peptide.

CONCLUSIONThe method of EOLA1 protein purification and renaturation was convenient and efficient, and by this method sufficient amount of highly pure EOLA1 protein could be obtained for the preparation of EOLA1 monoclonal antibody and for the study of its gene function.

Antibodies, Monoclonal ; Chromatography, Affinity ; Electrophoresis, Polyacrylamide Gel ; Endothelial Cells ; metabolism ; Humans ; Immunoblotting ; Membrane Proteins ; genetics ; isolation & purification ; metabolism

OBJECTIVETo measure the expression of phosphatidylinositol 3-kinase (PI3K) and protein kinase B (PKB) in liver tissue among low-birth-weight newborn rats treated with L-arginine (L-Arg) in early life, and to investigate the effect of L-Arg on insulin resistance.

METHODSEighteen pregnant rats were randomly divided into three groups: control, model and intervention (n=6 each). The control group was fed with normal protein feed (protein content=21%) during pregnancy to establish a normal-birth-weight newborn rat model, and the model and intervention groups were fed with low-protein feed (protein content=10%) during pregnancy to establish a low-birth-weight newborn rat model. Newborn rats from the three pregnant rat groups were also assigned to control, model and intervention groups. During 21 days of lactation, maternal rats in the control and model groups were fed with normal protein feed and normal drinking water, while maternal rats in the intervention group were fed with normal protein feed and drinking water rich in L-Arg (200 mg/kg·d). After ablactation, the three groups of newborn rats were fed with normal protein feed and normal drinking water. Liver tissue samples were collected from these newborn rats at 1, 3 and 8 weeks after birth. Protein expression of PI3K and PKB in liver tissue was measured by Western blot.

RESULTSAt 1 week after birth, the newborn rats in the intervention group had significantly higher protein expression of PI3K than in the model group (P=0.045), but there was no significant difference when compared with the control group (P=0.503). At 8 weeks after birth, the newborn rats in the intervention group had significantly higher protein expression of PKB than the model group (P=0.039), but there was no significant difference when compared with the control group (P>0.05).

CONCLUSIONSA supplement of L-Arg in early life can boost protein synthesis, increase protein expression of PI3K and PKB in liver tissue, promote insulin signaling and reduce insulin resistance.

Animals ; Animals, Newborn ; Arginine ; pharmacology ; Birth Weight ; Female ; Liver ; metabolism ; Male ; Phosphatidylinositol 3-Kinases ; genetics ; Phosphorylation ; Pregnancy ; Proto-Oncogene Proteins c-akt ; genetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the influence of integrin beta1 on the proliferation and differentiation of human keratinocyte stem cells (KSCs).

METHODSDNA oligonucleotides targeting integrin beta1 at different locations were synthesized and inserted into BamHI-1 HindIII linearized p Silencer 3.1/H1 plasmids. The inserted sequences were verified by DNA sequencing. The KSCs were divided into control (without transfection), T1 (with transfection of vacant vector), T2 (with transfection of si integrin beta(1-1) vector), T3 (with transfection of si integrin beta(1-1) vector), and T4 (with transfection of si Negative vector) groups. The change in the expression of integrin beta1, was determined with Western blotting. The positive vector with the highest expression of integrin beta1 was selected and named as integrin beta1, and semi-quantitative RT-PCR was employed to detect the change in the expression of integrin beta1 mRNA.

RESULTSThe protein expression of integrin beta1, was not suppressed in control and T1 group, but it was suppressed in T2 and T3 groups, especially in T3 group (the suppression rate was 60%-70%, which was named si integrin beta1). The expression of integrin beta1 mRNA was obviously decreased by integrin beta1, transfection (the suppression rate was 70%).

CONCLUSIONThe expression of integrin beta1, mRNA and protein could be down-regulated with recombinant si integrin P, vector transfection.

Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Genetic Vectors ; Humans ; Integrin beta1 ; genetics ; metabolism ; Keratinocytes ; metabolism ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Stem Cells ; metabolism ; Transfection