A cleaved adhesin DNA vaccine targeting dendritic cell against Porphyromonas gingivalis-induced periodontal disease.

BACKGROUND: Arg-gingipain A (RgpA) is the primary virulence factor of Porphyromonas gingivalis and contains hemagglutinin adhesin (HA), which helps bacteria adhere to cells and proteins. Hemagglutinin's functional domains include cleaved adhesin (CA), which acts as a hemagglutination and hemoglobin-binding actor. Here, we confirmed that the HA and CA genes are immunogenic, and using adjuvant chemokine to target dendritic cells (DCs) enhanced protective autoimmunity against P. gingivalis-induced periodontal disease. METHODS: C57 mice were immunized prophylactically with pVAX1-CA, pVAX1-HA, pVAX1, and phosphate-buffered saline (PBS) through intramuscular injection every 2 weeks for a total of three administrations before P. gingivalis-induced periodontitis. The DCs were analyzed using flow cytometry and ribonucleic acid sequencing (RNA-seq) transcriptomic assays following transfection with CA lentivirus. The efficacy of the co-delivered molecular adjuvant CA DNA vaccine was evaluated in vivo using flow cytometry, immunofluorescence techniques, and micro-computed tomography. RESULTS: After the immunization, both the pVAX1-CA and pVAX1-HA groups exhibited significantly elevated P. gingivalis-specific IgG and IgG1, as well as a reduction in bone loss around periodontitis-affected teeth, compared to the pVAX1 and PBS groups (p < 0.05). The expression of CA promoted the secretion of HLA, CD86, CD83, and DC-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) in DCs. Furthermore, the RNA-seq analysis revealed a significant increase in the chemokine (C-C motif) ligand 19 (p < 0.05). A notable elevation in the quantities of DCs co-labeled with CD11c and major histocompatibility complex class II, along with an increase in interferon-gamma (IFN-γ) cells, was observed in the inguinal lymph nodes of mice subjected to CCL19-CA immunization. This outcome effectively illustrated the preservation of peri-implant bone mass in rats afflicted with P. gingivalis-induced peri-implantitis (p < 0.05). CONCLUSIONS: The co-administration of a CCL19-conjugated CA DNA vaccine holds promise as an innovative and targeted immunization strategy against P. gingivalis-induced periodontitis and peri-implantitis.

Association Study of Mitochondrial DNA Haplogroup D and C5178A Polymorphisms with Chronic Kidney Disease.

Objective: To explore the associations of common mitochondrial DNA polymorphisms with chronic kidney disease (CKD). Methods: Data from 286 longevous individuals aged 95 years or older from the longevity arm from the Rugao Longevity and Ageing Study (RuLAS) were used. Twenty-eight common haplogroups defined by 33 single nucleotide polymorphisms were genotyped using SNaPshot minisequencing reaction assays. The creatinine-based estimated glomerular filtration rate (eGFR) was calculated using the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation. Results: The prevalence of CKD was 23.6% among the longevous participants aged 95 years and older. The D haplogroup (67.37 ± 14.72 vs. 70.65 ± 11.07, p = 0.045), the D5 haplogroup (60.86 ± 18.36 vs. 70.34 ± 11.53, p = 0.002), and the 5178A allele (67.23 ± 14.48 vs. 70.75 ± 11.10, p = 0.029) were associated with lower eGFR levels compared with noncarriers. The D5 haplogroup (13.8% vs. 3.6%, p = 0.005) was significantly higher, while D haplogroup (35.4% vs. 24%, p = 0.067) and the 5178A allele (36.9% vs. 24.9%, p = 0.056) were borderline significantly higher in CKD individuals than those without CKD. Further, after adjusting for multiple covariates, the D haplogroup, the D5 haplogroup, and the 5178A allele were associated with increased odds of CKD with odds ratios of 1.93 (95% confidence interval [CI]: 1.00-3.72, p = 0.050), 4.76 (95% CI: 1.49-15.22, p = 0.009) and 2.04 (95% CI: 1.05-3.96, p = 0.035), respectively. Conclusions: The D and D5 haplogroups, as well as the 5178A allele are associated with decreased eGFR levels and an increased risk of CKD in a longevous population.

Effect of chromodomain helicase/ATPase DNA binding protein 1-like gene on the invasion and metastasis of tongue squamous cell carcinoma CAL27 cells.

OBJECTIVES: A study was conducted to investigate the molecular mechanism of chromodomain helicase/ATPase DNA binding protein 1-like gene (CHD1L) influencing the invasion and metastasis of tongue squamous cell carcinoma and to provide a new target for clinical inhibition of invasion and metastasis of tongue squamous cell carcinoma. METHODS: Ualcan website was used to analyze the expression of CHD1L in normal epithelial tissue and primary head and neck squamous cell carcinoma and to analyze the effect of lymph node metastasis on the expression of CHD1L in tissues with head and neck squamous cell carcinoma. The relationship between CHD1L expression and the survival rate of patients with head and neck squamous cell carcinoma was tested by the GEPIA website. Western blot was used to quantify the levels of CHD1L protein in human tongue squamous cell carcinoma CAL27 and immortalized human skin keratinocyte cell HaCaT. After knocking down CAL27 in human tongue squamous cell carcinoma cells with an RNA interference plasmid, the cells were designated as SiCHD1L/CAL27 and Scr/CAL27. Western blot was utilized to detect the expression of CHD1L in each group of cells. The change in CAL27 cell proliferation ability was tested by EdU proliferation test after CHD1L knockdown. The change of cell migration ability of each group cells was tested through the wound healing assay. Western blot was used to detect epithelial-mesenchymal transition (EMT) marker E-cadherin and Vimentin protein expression levels. RESULTS: Ualcan database showed that the expression of CHD1L in primary head and neck squamous cell carcinoma tissues was higher than in normal epithelial tissues and in head and neck squamous cell carcinoma tissues with lymph node metastasis. GEPIA website analysis showed that the overall survival rate of patients with head and neck squamous cell carcinoma with high expression of CHD1L was significantly lower than that of patients with low expression. Western blot results showed that CHD1L expression in human tongue squamous carcinoma cells CAL27 was higher than that of human normal skin cells HaCaT. CHD1L expression in SiCHD1L/CAL27 cells was much lower than that in Scr/CAL27 cells. Results of EdU proliferation experiments showed the significant reduction in the cell proliferation ability of the SiCHD1L/CAL27 cells. Results of the wound healing experiments showed the reduction in the migration capacity of the SiCHD1L/CAL27 cells. The expression of E-cadherin increased, whereas that of Vimentin decreased, in SiCHD1L/CAL27 cells. CONCLUSIONS: CHD1L promoted the EMT, proliferation, migration, and invasion ability of tongue squamous cell carcinoma cells.

[Effects of RAB1A on the proliferation, invasion, and metastasis of tongue squamous cell carcinoma cells].

OBJECTIVE: This study aimed to investigate the molecular mechanism of RAB1A in the proliferation, invasion, and metastasis of human tongue squamous cell carcinoma. METHODS: Western blot was used to detect the expression of RAB1A protein in human normal tongue epithelial cells (Hacat) and tongue squamous cell carcinoma Tca8113. The changes in RAB1A after plasmid transfection were also studied. The Tca8113 cells were named SiRAB1A/Tca8113 after RAB1A plasmid transfection. The expression of the epithelial-mesenchymal transition (EMT)-related markers of SiRAB1A/Tca8113 cells was also detected. CCK-8 assay was used to detect the proliferation of SiRAB1A/Tca8113 cells. Transwell and wound healing assays were used to detect the invasive and metastatic abilities of SiRAB1A/Tca8113 cells, respectively. RESULTS: Western blot results showed that the expression of RAB1A in tongue squamous cell carcinoma cells was significantly higher than that in Hacat. RAB1A decreased significantly after SiRAB1A plasmid transfection. CCK-8 proliferation assay showed that the proliferation of SiRAB1A/Tca8113 cells also decreased significantly. Transwell and wound healing assays demonstrated that the invasive and metastatic abilities of SiRAB1A/Tca8113 cells decreased significantly, respectively. In addition, Western blot results demonstrated that RAB1A deletion significantly increased the expression of E-cadherin and inhibited the expression of Vimentin. CONCLUSIONS: RAB1A could promote the proliferation, invasion, and metastasis of tongue squamous cell carcinoma cells.

Associations of sleep quality and sleep duration with frailty and pre-frailty in an elderly population Rugao longevity and ageing study.

BACKGROUND: Previous studies suggest that poor sleep quality or abnormal sleep duration may be associated with frailty. Here we test the associations of sleep disturbances with both frailty and pre-frailty in an elderly population. METHODS: Participants included 1726 community-dwelling elders aged 70-87 years. Pittsburgh Sleep Quality Index (PSQI) was used to assess sleep disturbances. Frailty was defined using phenotype criteria. Logistic regression models were used to estimate odds ratio of the associations. RESULTS: The average PSQI score was 5.4 (SD, 3.1). Overall 43.6% of the participants had poor sleep quality (PSQI> 5), 8.2% had night sleep time ≤ 5 h, and 27.8% had night sleep time ≥ 9 h. The prevalence of frailty and pre-frailty was 9.2 and 52.8%, respectively. The proportions of PSQI> 5 increased with the severity of frailty status (robust: pre-frail: frail, 34.5%: 48%: 56.1%, P < 0.001). After adjustment for multiple potential confounders, poor sleep quality (PSQI> 5) was associated with higher odds of frailty (OR = 1.78, 95% CI 1.19-2.66) and pre-frailty (OR = 1.51, 95% CI 1.20-1.90). Sleep latency, sleep disturbance, and daytime dysfunction components of PSQI measurements were also associated with frailty and pre-frailty. In addition, sleep time 9 h/night was associated with higher odds of frailty and pre-frailty. CONCLUSIONS: We provided preliminary evidences that poor sleep quality and prolonged sleep duration were associated with being frailty and pre-frailty in an elderly population aged 70-87 years. The associations need to be validated in other elderly populations.

Preservation of level IIb lymph nodes during supraomohyoid neck dissection for clinically node-negative oral squamous cell carcinoma.

PURPOSE: To identify the significance of level IIb neck dissection for patients with clinically node-negative oral squamous cell carcinoma (OSCC). METHODS: A retrospective study was conducted with 203 patients with OSCC with no palpable lymph nodes in neck admitted to the Department of Oral Maxillofacial-Head and Neck Oncology from January 2012 through December 2014. After the diagnostic evaluations, all patients underwent wide local dissection and periodic supraomohyoid neck dissection (SOHND). In total, 115 patients underwent SOHND with IIb lymph node dissection, and 88 patients underwent elective SOHND without IIb lymph node dissection. The incidence of level IIb lymph node metastasis was evaluated by pathological and immunohistological analyses. The results were analyzed with independent sample t-tests. The incidence of complications (mainly scapular syndrome) and IIb lymph node metastasis rate (mainly for the preserving IIb group) were analyzed. RESULTS: In total, 7 (6.09%) of the 115 patients who underwent SOHND had level IIb lymph nodes involvement. After 3 years of follow-up, 83 (72.17%) patients who underwent SOHND had different degrees of scapular syndrome, and 27 (32.53%) patients who underwent SOHND improved through rehabilitation training but did not fully recover. Four (4.55%) patients who underwent elective SOHND (preserving IIb) developed scapular syndrome and recovered through rehabilitation after surgery. The 3-year overall survival rate of the 115 patients was 86.09%, and the 3-year overall survival rate of the 88 patients who underwent elective SOHND (preserving IIb) was 84.09%. There were no significant differences between the two groups (P > 0.05). CONCLUSION: Patients with clinically N0 OSCC have a low rate of level IIb lymph node metastasis. Level IIb lymph nodes resection are not necessary during SOHND, which thereby protects the accessory nerve and its branches from damage and improves patient quality of life.

Beneficial effect of 20­hydroxyecdysone exerted by modulating antioxidants and inflammatory cytokine levels in collagen­induced arthritis: A model for rheumatoid arthritis.

Rheumatoid arthritis is a chronic autoimmune disease characterized by an elevated synovial inflammatory response, with destruction or erosion of articular cartilage in major joints. The aim of the present study was to examine whether 20­hydroxyecdysone (HES) is able to ameliorate oxidative stress and inflammatory responses in a collagen­induced rheumatoid arthritis (CIA) rat model. A total of 40 healthy male rats were selected arbitrarily and separated into four groups. Rats treated with saline served as a control (group I), rats subjected to CIA induction by intradermal injection of bovine collagen II type served as the induced group (group II), while rats induced with CIA and administered with 10 and 20 mg/kg bodyweight HES for 28 days served as treatment groups (groups III and IV). Biochemical parameters, including paw swelling (edema), arthritis score, indexes of thymus and spleen, antioxidant levels (superoxide dismutase, catalase and glutathione), articular elastase and anti­collagen II specific immunoglobulins (Ig)G, IgG1 and IgG2a, in addition to inflammatory markers [nitric oxide, C­reactive protein, interleukin (IL)­1ß, IL­6, tumor necrosis factor­α and nuclear factor­κB p65 subunit] were significantly decreased (P<0.01) following supplementation with HES (10/20 mg/kg). Consistently, the protein expression pattern of inducible nitric oxide synthase and cyclooxygenease­2 were significantly downregulated (P<0.01) upon treatment with HES. In addition, histological analysis confirmed arthritis in CIA­induced rats by revealing the presence of greater polymorphonuclear cell infiltration, with eroded articular cartilage and prominent synovitis. However, administration of HES was demonstrated to alleviate the morphological changes and maintain the normal architecture of synovial joints. In conclusion, the results of the present study indicated that treatment with HES (particularly 20 mg/kg) may effectively eradicate the inflammatory cascade and oxidative stress process in CIA­induced rats and thereby exhibit anti­rheumatoid arthritis properties.

Diagnostic value of high-frequency color Doppler ultrasonography examination in combination with anti-cyclic citrullinated peptide antibody testing in rheumatoid arthritis patients.

We studied the diagnostic value of high-frequency color Doppler ultrasonography (HCDU) examination in combination with anti-cyclic citrullinated peptide (anti-CCP) antibody testing in rheumatoid arthritis (RA) patients with finger joint damage. From January 2015 to December 2015, 80 patients diagnosed with RA with finger joints damage were enrolled in this study. Patients were examined with HCDU. Serum anti-CCP antibody level was tested using ELISA, and results were compared with the healthy control group. Results obtained by ELISA demonstrated that the positive rate of anti-CCP antibodies was 73.8% in the study group, and 10% in the control group. The negative rate was 26.2% in the study group, and 90% in the control group. HCDU examination suggested that the predominantly affected joint by bone erosion of RA with finger joint damage was MCP3 (16.7%), followed by PIP3 (14.1%), MCP2 (13.5%) and PIP2 (12.8%). The slightest affected joint was thumb metacarpophalangeal joint, followed by thumb, little finger metacarpophalangeal joint and proximal interphalangeal joint. The sensitivity of diagnosis of RA with finger joints damage with both HCDU and CCP antibody examination showed a significantly lower level compared with examination with each one of the methods alone, while specificity showed a significantly higher level. Thus, a combination of HCDU examination and anti-CCP antibody testing can be considered useful to improve the early diagnostic rate of RA. HCDU examination is a sensitive, secure, atraumatic and easily-operated diagnostic method for early RA patients with finger joint damage. When combined with anti-CCP antibody testing, it will provide a better chance for RA patients, and give them hope for a better treatment and improved prognosis.

Correlation of nuclear factor-κB, regulatory T cell and transforming growth factor ß with rheumatoid arthritis.

Objective: Investigated the correlation of nuclear factor-κB, regulatory cells and transforming growth factor-ß with rheumatoid arthritis. Methods: Included 65 cases of RA patients admitted in our hospital from June 2015 to December 2016 into case group, and included 50 healthy people into control group during the same period. Collected the peripheral detection of nuclear factor-κB, regulatory cells and transforming growth factor beta levels, and compared them between two groups. Results: The percentage of CD4+, CD25+ T cells in the case group was significantly lower than that in the control group (P < .05); There was no significant difference in the percentage of CD4+, CD25+ CD127low/-, T cells between groups (P > .05); The levels of TGF - beta and NF - kappa B in the case group were higher than those in the control group, and the difference between the two groups was statistically significant (P < .05); The levels of ESR, CRP and RF in the case group were higher than those in the control group (P < .05). There was a negative correlation between the expression of nuclear factor-κB, transforming growth factor-ß and RF level in RA patients by pearson correlation analysis, r = -0.652, P < .05. Conclusion: The expression levels of CD4+, CD25+ T cells in patients with RA are significantly decrease, which has a negative correlation with RA activity index RF, and showed that the pathogenesis of RA is related to the regulation of immune system.

Diagnostic value of high-frequency ultrasound and magnetic resonance imaging in early rheumatoid arthritis.

Early diagnosis and management improve the outcome of patients with rheumatoid arthritis (RA). The present study explored the application of high-frequency ultrasound (US) and magnetic resonance imaging (MRI) in the detection of early RA. Thirty-nine patients (20 males and 19 females) diagnosed with early RA were enrolled in the study. A total of 1,248 positions, including 858 hand joints and 390 tendons, were examined by high-frequency US and MRI to evaluate the presence of bone erosion, bone marrow edema (BME), synovial proliferation, joint effusion, tendinitis and tendon sheath edema. The imaging results of the above abnormalities, detected by US, were compared with those identified using MRI. No statistically significant overall changes were observed between high-frequency US and MRI in detecting bone erosion [44 (5.1%) vs. 35 (4.1%), respectively; P>0.05], tendinitis [18 (4.6%) vs. 14 (1.5%), respectively; P>0.05] and tendon sheath edema [37 (9.5%) vs. 30 (7.7%), respectively; P>0.05]. Significant differences were observed between high-frequency US and MRI with regards to the detection of synovial proliferation [132 (15.4%) vs. 66 (7.7%), respectively; P<0.05] and joint effusion [89 (10.4%) vs. 52 (6.1%), respectively; P<0.05]. In addition, significant differences were identified between the detection of BME using MRI compared with high-frequency US (5.5 vs. 0%, respectively; P<0.05). MRI and high-frequency US of the dominant hand and wrist joints were comparably sensitive to bone erosion, tendinitis and tendon sheath edema. However, MRI was more sensitive in detecting bone marrow edema in early RA, while US was more sensitive in the evaluation of joint effusion and synovial proliferation. In conclusion, US and MRI are promising for the detection and diagnosis of inflammatory activity in patients with RA.

Expression and Significance of High-Mobility Group Protein B1 (HMGB1) and the Receptor for Advanced Glycation End-Product (RAGE) in Knee Osteoarthritis.

BACKGROUND This study was performed with the aim to explore the expression of high-mobility group protein B1 (HMGB1) and the receptor for advanced glycation end-product (RAGE) in knee osteoarthritis (KOA) and its clinical significance. MATERIAL AND METHODS A total of 108 synovial tissues selected from KOA patients were included in the experimental group. Seventy-five synovial tissues of knee joints, selected from patients who were clinically and pathologically confirmed without joint lesion, were included in the control group. The mRNA and protein expressions of HMGB1 and RAGE were determined by using RT-PCR and immunohistochemistry, respectively. Western blotting was used for measuring relative protein expression. An ROC curve was drawn to evaluate the diagnostic value of HMGB1 and RAGE for KOA. RESULTS The positive cell number and positive expression intensity of HMGB1 and RAGE in synovial tissue was higher in the experimental group than in the control group. PI for HMGB1 and RAGE expression in KOA patients was positively correlated with clinical classification of X-ray films (P<0.05). HMGB1 and RAGE mRNA expressions, as well as relative protein expression of HMGB1 and RAGE in synovial tissue, were higher in the experimental group than in the control group (all P<0.05). The sensitivity of HMGB1 protein, RAGE protein, HMGB1 mRNA, and RAGE mRNA were 76.9%, 64.8%, 86.1%, and 64.8%, respectively; and the specificity was 100%, 96%, 74.7%, and 80%, respectively. CONCLUSIONS The protein and mRNA expressions of HMGB1 and RAGE are both increased in KOA patients, suggesting that they are involved in KOA.

Evaluation of the wound-healing activity and anti-inflammatory activity of aqueous extracts from Acorus calamus L.

In folklore medicine, Acorus calamus has been used as a wound-healing agent for thousands of years; however, there have been few scientific reports on this activity so far. Now, we explored deeply the wound-healing effect of aqueous extracts from the fresh roots and rhizomes of A. calamus in vivo, as well as anti-inflammatory activity in vitro, so as to provide scientific evidence for the traditional application. The wound-healing effect was determined by the image analysis techniques and the histological analysis in the excisional wounding test, and the anti-inflammatory activity was evaluated by the real-time RT-PCR techniques in the lipopolysaccharide-induced RAW 264.7 cells test. Aqueous extracts, administered topically at the dose range from twice to thrice in a day, could enhance significantly the rate of skin wound-healing. Moreover, the extracts could effectively inhibit the mRNA expressions of inflammatory mediators induced by lipopolysaccharide in RAW 264.7 cells. These results showed significantly the wound-healing activity of aqueous extracts in the animal model of excise wound healing, and anti-inflammatory activity in vitro.

Leaf rolling controlled by the homeodomain leucine zipper class IV gene Roc5 in rice.

Leaf rolling is considered an important agronomic trait in rice (Oryza sativa) breeding. To understand the molecular mechanism controlling leaf rolling, we screened a rice T-DNA insertion population and isolated the outcurved leaf1 (oul1) mutant showing abaxial leaf rolling. The phenotypes were caused by knockout of Rice outermost cell-specific gene5 (Roc5), an ortholog of the Arabidopsis (Arabidopsis thaliana) homeodomain leucine zipper class IV gene GLABRA2. Interestingly, overexpression of Roc5 led to adaxially rolled leaves, whereas cosuppression of Roc5 resulted in abaxial leaf rolling. Bulliform cell number and size increased in oul1 and Roc5 cosuppression plants but were reduced in Roc5-overexpressing lines. The data indicate that Roc5 negatively regulates bulliform cell fate and development. Gene expression profiling, quantitative polymerase chain reaction, and RNA interference (RNAi) analyses revealed that Protodermal Factor Like (PFL) was probably down-regulated in oul1. The mRNA level of PFL was increased in Roc5-overexpressing lines, and PFL-RNAi transgenic plants exhibit reversely rolling leaves by reason of increases of bulliform cell number and size, indicating that Roc5 may have a conserved function. These are, to our knowledge, the first functional data for a gene encoding a homeodomain leucine zipper class IV transcriptional factor in rice that modulates leaf rolling.

[A clinical analysis of hemophagocytic syndrome in autoimmune diseases].

OBJECTIVE: To analyze the clinical features of patients with hemophagocytic syndrome (HPS) in autoimmune diseases (AID). METHODS: We collected the data of 11 patients with AID complicated with HPS in Peking Union Medical College Hospital from 2004 to 2009. The underlying diseases, clinical features, laboratory findings and treatment outcomes were retrospectively analyzed. RESULTS: Of the 11 patients, 3 were male, 8 were female. Mean age was (30.7 ± 18.3) years. The underlying diseases included Still disease (n = 4), systemic lupus erythematosus(n = 3), and rheumatoid arthritis, primary Sjögren's syndrome, Wegener granulomatosis and Crohn disease in each one case. HPS was associated with the onset of AID (n = 4), active infection alone (n = 1) and both factors (n = 6). HPS was clinically characterized by high fever (100%), hepatosplenomegaly (72.7%), lymphadenopathy (63.3%) and central nervous system involvement (36.3%). 4 patients presented with disseminated intravascular coagulation (DIC) (36.3%). Laboratory data mainly manifested with cytopenia (100%), liver dysfunction (100%), hypofibrinogenemia (62.5%), hypertriglyceridemia (81.8%), serum ferritin > 500 µg/L (100%), low NK-cell activity (80%) and hemophagocytosis in bone marrow (100%). Based on treating underlying infections and use of corticosteroids and immunosuppressive agents in combination with intravenous immunoglobulins (IVIG) therapy, 5 patients recovered, 6 patients died. The mortality rate was 54.5%. DIC were associated with mortality (r = 0.69, P = 0.019). CONCLUSION: The episode of HPS always occurs simultaneously with multiple system involvement that was often difficult to distinguish from active AID. The present of DIC on HPS related with poor prognosis and high mortality. Corticosteroids and immunodepressant and IVIG may improve the prognosis of HPS, while anti-infection therapy is very important and necessary for the patients accompany with active infection.

[Tissue distribution of solid lipid nanoparticles loaded garlic oil in rats].

OBJECTIVE: To investigate the tissue distribution of the diallyl disulfide (DADS) and diallyl trisulfide (DATS) in solid lipid nanoparticles loaded garlic oil (GO-SLN) in rats. METHOD: The gas chromatography-electron capture detection (GC-ECD) method was established to determined the DADS and DATS simultaneously in the biological samples of rats after administration of 0.5 mL garlic oil injection or GO-SLN (containing about 10 mg garlic oil) via jugular vein cannula. The conditions for gas chromatographic separation were as follows. The oven temperature was set at 110 degrees C and maintained for 15 min. Temperatures at the injection port and detector were 180 degrees C and 300 degrees C, respectively. Ultra-pure nitrogen (purity > 99.999%, Shenyang Kerui Special Gases Co. Ltd., China) was used as a carrier gas and made-up gas at flow-rates of 1 mL x min(-1) and 60 mL x min(-1), respectively. All injections were carried out in the split injection mode with a split ratio of 1:10. RESULT: The GC-ECD method was fit for determing the concentration of DADS and DATS in garlic oil. The distribution character of GO-SLN in rats had changed to some extent and the concentration of GO-SLN in tissues was higher than that of GO-Injection. CONCLUSION: The SLN can elevate the passive targeting of drugs and lengthen their action time in tissues.

[Recombinant fowlpox virus coexpressing HA and NA gene from subtype H5N1 of avian influenza virus and its protective efficacy].

The hemagglutinin (HA) and neuraminidase(NA)gene from subtype H5N1 avian influenza virus were directly inserted into the transferring vector pllS, resulting in the recombinant transferring vector p11SH5ANA. Then p11SH5ANA was transfected into the chicken embryo fibroblasts (CEF), which was pre-infected with wild type fowlpox virus, to generate the recombinant fowlpox virus coexpressing H5A and NA (rFPV-11SH5ANA). By selection of blue plaques on the CEF, rFPV-11SH5ANA was obtained and purified. Experiments on SPF chickens demonstrated that the HI antibody titers in chickens vaccinated with HA-NA coexpressed vaccine was higher than those with HA expressed monovalent vaccines, and all chickens receiving either rFPV-11SH5ANA or rFPV-11SH5A were completely protected from the virulent AIV(H5N1) challenge, while those receiving wt-FPV experienced 100% mortality. The results showed that the rFPV-11SH5ANA was a safe and highly efficient gene engineering vaccine candidate for preventing HPAI.

[Studies on diclofenac sodium pulsatile release pellets].

AIM: To investigate the preparation of diclofenac sodium pulsatile release pellets (DS-PRP), the release in vitro and the pharmacokinetics of the drug. METHODS: Diclofenac sodium (DS) core pellets prepared by extrusion-spheronization technology were coated in a mini-fluidized bed spray coater with swelling material as the inner coating swelling layer and ethylcellulose aqueous dispersion as the outer coating controlled layer. The effects of formulation and medium on pulsatile release of DS were investigated under release rate test. Pharmacokinetic and bioavailability study in eight human subjects were performed by HPLC method. RESULTS: The delayed-release time and release rate of DS from DS-PRP were influenced obviously by the swelling material, the concentration of SDS in medium, the coating level of the inner swelling layer and the outer controlled layer. In vitro, the delayed-release time T0.1 was 3.1 h, and the pulsed-release time T0.1-0.2 was 1.2 h. In vivo, the delayed-release time Tlag was 2.8 h, and the bioavailability was (91 +/- 12)%. CONCLUSION: The release of drug from DS-PRP was shown to be in pulsed way both in vitro and in vivo.