The activities of interleukin 6 (IL 6) and tumor necrosis factor alpha (TNF?) produced by peripheral blood mononuclear cells (PBMCs) in vitro and their relevant levels in sera were measured in 49 patients with psoriasis vulgaris (PV) and in 20 normal controls using bioassay. The relationship between them was also analysed. The results showed that in PV patients the activites of IL 6 and TNF? produced by PBMCs and their serum levels were higher than those in healthy controls. There was a positive linear correlation between the activites in vitro and relevant levels in sera, and a positive linear correlation between serum levels of IL 6 and TNF?. These results indicate that there is dysfunction of cellular immune of PBMCs in PV patients. The increase of serum levels of IL 6 and TNF? may be related to the increased activity of PBMCs, and there may be a synergic action between IL 6 and TNF? in the pathogenesis of psoriasis.

BACKGROUND:Studies have found that combination of two of chitosan (CS), nano-hydroxyapatite (nHA) and poly(lactide-co-glycolide) (PLGA) can improve the mechanical properties and biocompatibility of the composite stent in certain extent as wel as improve osteogenic differentiation of the cels, but there is a certain distance from the ideal bone tissue engineering scaffolds.
OBJECTIVE:To study biocompatibility and osteoinductive activity of nHA/CS/PLGA scaffolds with different proportions in vitro.
METHODS: nHA/CS/PLGA scaffolds were prepared at mass ratio of 10:10:80, 10:20:70, 20:10:70 respectively by particle leaching method. And human bone marrow stem cels (hBMSCs) were co-cultured with these scaffolds in vitro. Adhesion, proliferation, and osteoinductive activity of these scaffolds were examined qualitatively and quantitatively by growth curve of hBMSCs on scaffolds. Gene expression of alkaline phosphatase activity and osteocalcin was detected by RT-PCR.
RESULTS AND CONCLUSION: hBMSCs could be attached, proliferated, and osteoinduced better on the nHA/CS/PLGA scaffold with the mass ratio of 20:10:70, compared to the other two groups of scaffolds. The differences were significant statisticaly (P< 0.05). Alkaline phosphatase and osteocalcin expressions were respectively higher in the scaffold with the mass ratio of 20:10:70 after 9-27 days of co-culture and 15-27 days of co-culture, in comparison with the other two groups of scaffolds. These findings indicate that the nHA/CS/PLGA scaffolds with the mass ratio of 20:10:70 demonstrated preferable biocompatibility and osteogenic inductivity, which is expected to be a promising scaffold material for bone tissue engineering.

Objective To study multimodal analgesia in patients who underwent transarterial chemoembolization (TACE) for hepatocellular carcinoma (HCC).Methods 60 patients who underwent TACE for HCC from Aug.2016 to Nov.2016 were randomized into two groups:the multimodal analgesia group and the control group.The pain scores of these two groups of patient during the procedure and at different posttreatment time points,and the rates of adverse effect and pharmacoeconomic differences were recorded.Results When compared to the control group,the pain scores at 0 h,2 h,4 h,6 h,12 h after treatment in the multimodal analgesia group were significantly lower (P ＜ 0.05),and the satisfactory scores for the patients were significantly improved (96.6％ vs.66.7％).The multimodal group of patients also had significandy lower adverse effect rates of nausea and vomiting,and it was more cost-effective.Conclusions Patients who required multimodal analgesia had better pain relieve,patient satisfaction and less adverse reactions after TACE than patients in the control group.Multimodal analgesia was a safe,effective and economic way to control TACE pain and it was worth recommended in clinical practice.

The study aims to establish a method for simultaneous determination of repaglinide and pravastatin sodium in rat plasma by LC-MS/MS and to study its pharmacokinetic interactions. Eighteen male SD rats were divided into repaglinide group, pravastatin sodium group and co-administration group. Blood samples were collected at different times after oral administration. Repaglinide and pravastatin sodium in rat plasma were separated by Agilent HC-C18 with the mobile phase consisting of methanol-0.1% formic acid (80 : 20). Detection and quantification were performed by using ESI-MS. The detector was operated in selected Reaction-monitoring mode at m/z 453.3-->230.1 for repaglinide, m/z 447.2-->327.4 for pravastatin sodium and m/z 285.1-->192.9 for diazepam as the internal standard. The calibration curve obtained was linear (R2>0.99) over the concentration range of 9.77-10,000 ng.mL-1 for repaglinide and 4.88-625 ng.mL-1 for pravastatin sodium. Compared with the single administration group, Cmax and AUC0-6h of repaglinide increased significantly (P<0.05) and tmax of pravastatin sodium prolonged (P<0.05) in co-administration group. The method is found to be simple, sensitive and accurate for determining the concentration of repaglinide and pravastatin sodium in rat plasma. There exists pharmacokinetic interactions in the co-administration of repaglinide and pravastatin sodium.
Administration, Oral ; Animals ; Carbamates ; administration & dosage ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; Drug Interactions ; Male ; Piperidines ; administration & dosage ; blood ; pharmacokinetics ; Pravastatin ; administration & dosage ; blood ; pharmacokinetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Sensitivity and Specificity ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

OBJECTIVETo explore effects of acupuncture on ability of daily living (ALD) and the incidence rate of disability and mortality of the patient of acute ischemic stroke.

METHODSForty patients with acute ischemic stroke were randomly assigned to an acupuncture group and a control group, 20 cases in each group. The treatment group were treated with acupuncture for 3-4 weeks, 5 times each week, and routine therapy. The control group were treated with routine therapy alone.

RESULTSNo statistically significant differences between the two groups in the score of neurological defection, and the incidence rate of disability and mortality at following survey of 3 and 6 months were found.

CONCLUSIONAcupuncture is safe and feasible for stroke at early stage.

Acupuncture ; Acupuncture Therapy ; Humans ; Stroke ; therapy

OBJECTIVETo screen a variety of Porphyromonas gingivalis (Pg) common outer membrane proteins with two-dimensional liquid phase fractionation (PF2D) and matrix-assisted laser desorption ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF-MS) and provide candidate target antigen for the design of vaccines with cross protection against a variety of Pg.

METHODSThe outer membrane proteins of Pg301, PgATCC33277 and PgW83 were extracted through ultracentrifugation, and then they were separated by ProteomeLab PF2D protein fractionation system. After separation, the outer membrane proteins were obtained through comparison, and the primary structure of the proteins was identified by MALDI-TOF/TOF-MS and database.

RESULTSNinety-nine protein samples out of 3 strains of Pg were obtained after the high performance chromato focusing (HPCF) separation process. B7 fractions of 3 strains of Pg were separated by the reversed-phase high performance liquid chromatography (RP-HPCL) separation process. After comparison of peak and retention time of chromatogram, the 8 common protein peaks of 3 strains of Pg were confirmed. The protein samples were identified by MALDI-TOF/TOF-MS, and one of them was known protein arg-gingipain A.

CONCLUSIONSPF2D protein fractionation system is of good reproducibility and high resolution. A combination of PF2D and MALDI-TOF/TOF-MS can be used to identify the common outer membrane proteins of Pg.

Antigens, Bacterial ; analysis ; Mass Spectrometry ; Membrane Proteins ; analysis ; Porphyromonas gingivalis ; immunology ; Reproducibility of Results ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Vaccines

OBJECTIVETo clone the catalytic domain gene sequence of RgpAcd of Porphyromonas gingivalis (P. gingivalis) and to induce its fusion expression in E. coli.

METHODSThe desired DNA fragment RgpAcd was obtained by PCR and was separately sequenced and identified by inserting into inter-vector pMD18-T vector. The correctly fragment was linked with and cloned into a prokaryotic expression vector pET-15b. The recombinant expression plasmid which had been confirmed by enzymes digestion was transformed to E. coli competent cells BL21 (DE3) and expression of fusion protein was induced by IPTG.

RESULTSA 1 476 bp specific fragment was obtained and DNA sequencing showed that the fragment was consistent with those of the published. After induction with IPTG, a fusion protein of 5 x 10(4) was visualized on SDS-PAGE gel.

CONCLUSIONThe protein of RgpAcd will be obtained for further study and its protein was correctly expressed in E. coli BL21 cells.

Cloning, Molecular ; Cloning, Organism ; Escherichia coli ; Genetic Vectors ; Polymerase Chain Reaction ; Porphyromonas gingivalis ; Recombinant Fusion Proteins ; Recombinant Proteins

OBJECTIVETo study the clinicopathologic features of follicular dendritic cell sarcoma (FDCS) and its differential diagnosis.

METHODSTen cases of FDCS were studied by light microscopy, immunohistochemistry and in-situ hybridization. The clinical features and follow-up information were analyzed.

RESULTSAmongst the 10 cases of FDCS studied, the male-to-female ratio was 1:1. The mean age of the patients was 42 years. Six of them were located in cervical and peritoneal lymph nodes and four in extranodal sites (including tonsil, pelvic cavity, tail of pancreas and spleen). Histologically, the tumor cells had whorled, storiform or diffuse growth patterns. They were spindle in shape and contained syncytial eosinophilic cytoplasm, with round or oval nuclei, vesicular chromatin, distinct nucleoli and a variable number of mitotic figures. Multinucleated tumor giant cells and intranuclear pseudoinclusions were occasionally seen. There was a sprinkling of small lymphocytes and neutrophils within the tumor as well as in the perivascular region. Immunohistochemical study showed that the tumor cells were diffusely or focally positive for CD21, CD23, CD35 and D2-40, but negative for LCA, CD20, CD3, CD1a, HMB45 and CK. Some of them showed EMA, CD68 and S-100 reactivity. In-situ hybridization for Epstein-Barr virus-encoded RNA (EBER) showed positive signals in only one case (which was diagnosed as inflammatory pseudotumor-like FDCS). Of the 7 patients with follow-up information available (duration: 2 months to 39 months; mean: 14 months), 2 cases with paraneoplastic pemphigus died of pulmonary infection at 5 and 7 months respectively. The remaining 5 patients were alive and disease-free after surgical excision (+/- chemotherapy and radiotherapy).

CONCLUSIONSFDCS is a rare low to intermediate-grade malignant tumor. Appropriate application of FDC markers, such as CD21, CD35 and D2-40, would be helpful for arriving at a correct diagnosis. Most cases are associated with good prognosis after surgical treatment, with or without chemotherapy and radiotherapy. Patients with paraneoplastic pemphigus carry a less favorable prognosis.

Adult ; Antibodies, Monoclonal, Murine-Derived ; metabolism ; Dendritic Cell Sarcoma, Follicular ; complications ; metabolism ; pathology ; surgery ; Dendritic Cell Sarcoma, Interdigitating ; pathology ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Lymph Node Excision ; Lymph Nodes ; pathology ; surgery ; Male ; Meningioma ; pathology ; Middle Aged ; Nasopharyngeal Neoplasms ; pathology ; Paraneoplastic Syndromes ; complications ; Pemphigus ; complications ; Receptors, Complement 3b ; metabolism ; Receptors, Complement 3d ; metabolism ; Receptors, IgE ; metabolism ; Tonsillar Neoplasms ; metabolism ; pathology ; surgery ; Young Adult

Recombinant human interferon α2b(rhIFNα2b)is widely used as an antiviral therapy agent for the treatment of hepatitis B and hepatitis C.The current identification test for rhIFNα2b is complex.In this study,an anti-rhIFNα2b nanobody was discovered and used for the development of a rapid lateral flow strip for the identification of rhIFNα2b.RhIFNα2b was used to immunize an alpaca,which established a phage nanobody library.After five steps of enrichment,the nanobody I22,which specifically bound rhIFNα2b,was isolated and inserted into the prokaryotic expression vector pET28a.After subsequent purification,the physicochemical properties of the nanobody were determined.A semiquantitative detection and rapid identification assay of rhIFNα2b was developed using this novel nanobody.To develop a rapid test,the nanobody I22 was coupled with a colloidal gold to produce lateral-flow test strips.The developed rhIFNα2b detection assay had a limit of detection of 1 μg/mL.The isolation of I22 and successful construction of a lateral-flow immunochromatographic test strip demonstrated the feasibility of performing ligand-binding assays on a lateral-flow test strip using recombinant protein products.The principle of this novel assay is generally applicable for the rapid testing of other com-mercial products,with a great potential for routine use in detecting counterfeit recombinant protein products.

OBJECTIVETo clone the glyceraldehydes 3-phosphate dehydrogenase (GAPDH) gene of Porphyromonas gingivalis (P. gingivalis) and to induce its fusion expression in E. coli.

METHODSGAPDH was obtained by PCR and was inserted into cloning vector pMD-18-T to construct clone recon. Double enzymes digest the recon pMD18-T-GAPDH and the prokaryotic expression vector pET-32a and then connect to get the expressing recon pET-32a-GAPDH. The recombinant expression plasmid which had been confirmed by enzymes digestion was transformed to E. coli competent cells BL21 and induced the expression of GAPDH with isopropyl beta-D-1-thiogalactopyranoside (IPTG) of different density.

RESULTSDNA sequencing showed that the fragment was 99.802% the same to the sequence published in NCBI. Under the best density, IPTG could be highly expressed.

CONCLUSIONThe GAPDH had been successfully cloned and expressed in E. coli which gets ready for the following experiment to study the immunity of GAPDH and the homologues antibody preparation.

Cells, Cultured ; Cloning, Molecular ; Cloning, Organism ; Escherichia coli ; Genetic Vectors ; Glyceraldehyde ; Oxidoreductases ; Phosphates ; Polymerase Chain Reaction ; Porphyromonas gingivalis