OBJECTIVETo observe the biocompatibility of new biomaterials porous calcium phosphate (CPC) and ectopic bone formation of CPC with bone marrow stromal cells (BMSCs).

METHODSThe BMSCs were cultured from Beagle dog and combined with the porous CPC with the best concentration after transfect green fluorescent protein (GFP). The adhesion and growth of BMSCs on CPC were observed under inversion, fluorescence and scanning electron microscopy. The ectopic bone formation were observed at the 8th week after CPC and BMSCs were implanted subcutaneously into nude mice.

RESULTSWhen BMSCs with CPC were cultured at the 1st day, cells were climbing out from CPC with normal morphology. At the 7th day cells can be seen protruding pseudopods, secretion of matrix. Bone formation could be seen histomorphologically at the 8th week.

CONCLUSIONPorous CPC has good biocompatibility and is an ideal scaffold material for bone tissue engineering.

Animals ; Biocompatible Materials ; Bone Cements ; Bone and Bones ; Calcium Phosphates ; Dental Cementum ; Dogs ; Mesenchymal Stromal Cells ; Mice ; Mice, Nude ; Tissue Engineering

OBJECTIVETo investigate the effects of Qingre Huoxue Decoction , clearing heat and promoting blood flow; QRHXD), on the radiographic progression in patients with rheumatoid arthritis (RA) by X-ray imaging.

METHODSEighty-six patients with active RA diagnosed as damp-heat and blood stasis syndrome were randomized into a QRHXD group and a QRHXD plus methotrexate (MTX) group, with 43 cases in each group. After one-year of treatment, 21 cases in each group (42 in total) were evaluated. Radiographs of hands were obtained at the baseline and after 12 months of treatment. Images were evaluated by investigators blinded to chronology and clinical data, and assessed according to the Sharp/Van der Heijde methods.

RESULTSHigh intrareader agreements were reached (mean intraobserver intraclass coefficients: 0.95). No significant change in any imaging parameters of joint destruction was observed at 12 months in either group; and the differences between the two groups were not significant (P>0.05). The mean of the changing score in the QRHXD group was 3.5 ± 4.1, and 2.4 ± 3.5 in the QRHXD+MTX group, while the baseline radiographic score of patients in the QRHXD group was relatively higher (18.9 ± 19.1 vs. 14.0 ± 14.0). The mean rates of the changing scores of the two groups were similar (0.24 ± 0.28 vs. 0.25 ± 0.44, P=0.40). The severity of progression in the two groups was also similar (P=0.46), 7 cases without radiographic progression in the QRHXD group and 8 in the QRHXD+MTX group, 3 cases with obvious radiographic progression in the QRHXD group and 1 in the QRHXD+MTX group.

CONCLUSIONRadiographic progression of RA patients in both groups is similar, indicating that the QRHXD Decoction has a potential role in preventing bone destruction.

Adult ; Arthritis, Rheumatoid ; diagnostic imaging ; drug therapy ; Disease Progression ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Intention to Treat Analysis ; Male ; Methotrexate ; therapeutic use ; Middle Aged ; Radiography

OBJECTIVETo investigate the effect of Qingre Huoxue decoction, (QRHX) on radiographic progression in patients with rheumatoid arthritis (RA) with X-ray imaging.

METHODSFrom July 2007 to March 2009, 86 patients with active RA who were diagnosed as damp-heat and blood stasis syndrome were randomly divided into QRHX group and QRHX plus methotrexate (MTX) group, 43 cases in each group. Patients in the QRHX group were treated with QRHX decoction [composed of Huangbai, Chishao, Bixie, Danshen, Ezhu, Qingfengteng, raw Huangqi, Jinyinhua, Tufuling, Wugong, Fengfang, raw Yiyiren, which was cooked with water as 400 ml liquid); while patients in the other group were treated with QRHX decoction plus MTX. After one-year observation, 21 patients in each group (42 in total) were evaluated,with 19 females in QRHX group, average age of (43.0 +/- 11.3) years, and the course of the disease was 2 (1,3) years; and 18 females in QRHX+MTX group, average age of (44.5 +/- 14.0) years, and the course of the disease was 3 (1.7, 5) years. Radiographs of hands were obtained at baseline and 12 months after treatment. Images were evaluated by investigators blinded to chronology and clinical data, and assessed according to Sharp/van der Heijde methods.

RESULTSHigh intrareader agreements were reached (mean intraobserver intraclass coefficients: 0.95). No significant change in any imaging parameters of joint destruction was observed at 12 months after treatment in both groups; and there were no statistical differences between the two groups (P > 0.05). The severity of progress in two groups was also similar (P = 0.46), 7 patients without radiographic progress in QRHX group and 8 patients in QRHX+MTX group,3 patients with obvious radiographic progress in QRHX group and 1 patient in QRHX+MTX group.

CONCLUSIONRadiographic progress of RA patients in two groups is similar, indicating QRHX decoction has a potential role in preventing bone destruction.

Adult ; Arthritis, Rheumatoid ; diagnostic imaging ; drug therapy ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Methotrexate ; therapeutic use ; Middle Aged ; Radiography

Objective To study the effects of CpG oligodeoxyribonuclentide (ODN)as adjuvant on the immune responses in PBMC and CD4+ T cell with chronic hepatitis B virus. Methods The selected 20 infections were averagely divided two groups. The frequency of IFN-γ secreting PBMC and CD4+ T cell in immune tolerant phase and in the immune clearance phase that had stimulated by CpG ODN, HBsAg and Mixture[CpG ODN+HBsAg] were analyzed by enzyme linked immune spot(ELISOT). Results The PBMC and CD4+ T cell were differently incubated by CpG ODN+ HBsAg and M[CpG ODN + HBsAg]. The number of IFN-γ spot differently are 3±8, 339±429, 375±496, 1±4, 5±16 and 5±12;the results of immume tolerance are 3±8,361±153, 375±276, 0±2, 2±2 and 4±4; but the results of immune clearance are 3±21, 289±345, 405±656, 2±14, 8±40 and 7±30. The IFN-γ spots statistical analysis of PBMC were differently incubated by HBsAg and M,the total is P = 0.720, The IFN-γ spots statistical analysis of CD4+ T cell were differently incubated by HBsAg and M, the total is P = 0. 890, The IFN-γ spots statistical analysis of PBMC and CD4+ T cell were differently incubated by M, the total is P=0.000. Conclusions The ability that CpG ODN can not significantly increase the IFN-γsecreting of PBMC and CD4+ T cell that were incubated by HBsAg to the infection in immune tolerant phase and in the immune clearance phase, but the PBMC outweighed The CD4+ T cell.

Objective To investigate the clinical and pathological features, diagnosis and treatment of Castleman disease (CD).Methods Clinical features and related information on diagnosis and treatment of eight cases of CD were retrospectively analyzed.The size of involved lymph nodes ranged from (2 cm×2 cm×3 cm)-(4 cm×3 cm×2 cm).The lymph nodes were found in level Ⅰ (1 case), level Ⅱ (3cases) ,level Ⅲ (3 cases) and level Ⅳ (1 case).CT examination in eight patients showed the lesions manifested as ellipse soft masses.Dynamic contrast CT scan in four patients showed ring-enhanced area around the masses.Blood routine examination in eight patients were generally normal, with six patients had mild anemia.Results Based on the clinical classification, all lesions in this group were localized CD.Histopathology indicated that all lesions were of hyaline-vascular type. After surgery, there was no recurrence during the follow-up period. ConclusionsPatients with localized CD mainly have lymphadenectasis in a single location.The CT scan can give some evidence.Surgery should be given first priority.

Objective To investigate the expression of vasoactive intestinal peptide (VIP)in gastric adenocarcinoma,and to evaluate the correlation of VIP level with clinical pathologic parameters. Methods The level of VIP in sera from gastric adenocarcinoma patients and healthy people was investigated by ELISA. Moreover,the differential gene expression between gastric adenocarcinoma,gastric dysplasia,and the corresponding normal gastric mucosa were determined by RT-PCR. Western Blot was also used to measure the expression of VIP in the gastric odenouarcinoma and the normal gastric mucosa. Results The serum level of VIP was (5.794±0.014) ng/ ml in normal control and was (14.437±0.825) ng/ml in gastric adenocareinoma patients,showing significant difference (P<0.05). Meanwhile,the V/B of gastric adenocarcinoma tissues was greater than that of gastric dysplasia and the corresponding normal gastric mucosa (P < 0.01 ),the values of V/B were 1.5261±0.3028,0.9334±0.2872,and 0.9051±0.2794,respectively. The values of V/B between normal gastric mncesa and gastric dysplasia were not different significantly (P0.05).There were significantly negative correlation between the VIP mRNA expression of the differentiation degree of tumor( P < 0.05). The VIP mRNA expression was higher in gastric adenocarcinoma with lymph node metastasis than that without lymph node matsstsis (P<0.05).The VIP protein expression of the gastric adenouarcinoma tissues was greater than that of normal control. Conclusion This findings provide a direct evidence to support the possibility that VIP play a cofactor role in the pathogenesis of gastric adenccareinoma.

OBJECTIVETo investigate the change in T cell-mediated immunity and its relationship with plasma high mobility group box-1 protein (HMGB1) levels in severely burned patients.

METHODSThirty-five extensively burned patients (> 30% total body surface area) were included in this study, and were divided into MODS group (n = 13) and non-MODS group (n = 22). The blood samples were collected on post burn days 1, 3, 5, 7, 14, 21 and 28. The plasma levels of HMGB1 were measured by using ELISA, and T lymphocyte proliferation response and its IL-2 production ability in peripheral blood were determined too. In addition, the ratio of CD4+/CD8+ T cells were detected by using flow cytometry.

RESULTSPlasma HMGB1 levels were markedly elevated on post burn day 1 in severely burned patients, and HMGB1 level was significantly higher in MODS group than in non-MODS group (P < 0.05). Lymph proliferation response and IL-2 production of T cells in peripheral blood, and the ratio of CD4+/CD8+ T cells in MODS group were markedly lower than those in non-MODS group on post burn days 1, 14, 21 and 28 (all P < 0.05). It indicated that plasma HMGB1 levels were negatively correlated to T cellular immune function parameters, including lymphocyte proliferation response, IL-2 production, and the ratio of CD4+/ CD8+ T cells in extensively burned patients (all P < 0.05).

CONCLUSIONSExtensive burns could lead to T cellular immune dysfunction, which appears to be associated with the development of MODS. HMGB1, as an important late mediators of inflammation, might be involved in the pathogenesis of suppression of T cell-mediated immunity in these patients.

Adolescent ; Adult ; Burns ; blood ; complications ; immunology ; Female ; HMGB1 Protein ; blood ; Humans ; Immunity, Cellular ; immunology ; Male ; Middle Aged ; Multiple Organ Failure ; etiology ; immunology ; T-Lymphocytes ; immunology

AIMA series of triazole antifungal agents were synthesized to search for novel triazole antifungal agents with more potent activity, less toxicity and broader spectrum.

METHODSTwenty-one 1-(1H-1, 2, 4-triazolyl)-2-(2, 4-diflurophenyl)-3-(4-substituted-1-piperazinyl)-2-propanols were synthesized, on the basis of the three dimensional structure of P450 cytochrome 14alpha-sterol demethylase (CYP51) and their antifungal activities were also evaluated.

RESULTSResults of preliminary biological tests showed that most of title compounds exhibited activity against the eight common pathogenic fungi to some extent and the activities against deep fungi were higher than that against shallow fungi. In general, phenyl and pyridinyl analogues showed higher antifungal activity than that of the phenylacyl analogues.

CONCLUSIONSeveral title compounds showed higher antifungal activities than fluconazole and terbinafine. Compound VIII-1, 4, 5 and IX-3 showed the best antifungal activity with broad antifungal spectrum and were chosen for further study.

Antifungal Agents ; chemical synthesis ; chemistry ; pharmacology ; Aspergillus fumigatus ; drug effects ; Candida albicans ; drug effects ; Cryptococcus neoformans ; drug effects ; Fluconazole ; pharmacology ; Microbial Sensitivity Tests ; Molecular Structure ; Naphthalenes ; pharmacology ; Structure-Activity Relationship ; Triazoles ; chemical synthesis ; chemistry ; pharmacology

OBJECTIVETo apply pulse-field gel electrophoresis analysis(PFGE) in the analysis of cholera outbreak events and to determine the molecular epidemiological characteristics of Vibrio cholerae ( V. cholerae) isolates.

METHODSPFGE using restriction enzyme Not I was employed in the molecular subtyping of forty-one strains of V. cholerae isolated in cholera outbreak events from 2003 to 2005 in Guangzhou area and PFGE patterns were analyzed by BioNumerics Version 4.0 software to perform cluster analysis. Pattern profiles were compared by utilizing of Dice coefficient and UPGMA(unweighted pair group method with arithmetic averages). Comparison of PFGE typing results was performed with phage-biological typing and pathogenicity-associated genes typing.

RESULTSIn cholera outbreak events, PFGE could discriminate epidemiologically related and unrelated strains, having more discriminatory power than phage-biological typing and pathogenicity-associated genes-typing.

CONCLUSIONSMolecular sub-typing by PFGE could disclose the epidemiological relationships of strains from humans and the environment, providing molecular epidemiological evidence and support for the source-tracking of cholera outbreak events.

Bacterial Typing Techniques ; methods ; Cholera ; epidemiology ; microbiology ; Disease Outbreaks ; Electrophoresis, Gel, Pulsed-Field ; Humans ; Molecular Epidemiology ; Vibrio cholerae ; classification ; genetics ; isolation & purification

The purpose of this study was to construct 4 types of nonsense-mutated eukaryotic expression plasmids of fIX gene, using pcDNA3.1 plasmid containing fIX cDNA as template, and to identify, then to perform their expression in COS-7 cells. These stop mutants constructed by site-directed mutagenesis based on PCR, and further confirmed by DNA sequencing. COS-7 cells were transfected with either the wild-type or mutated fIX expression constructs, then the relative expression levels of fIX mRNA were detected by real time fluorescent quantitative PCR. The result showed that except the designed sites, there were no other nucleotide mutation in the sequences of four nonsense mutants. The results of real time PCR proved that the nonsense-mutated vectors can be effectively expressed in COS-7 cells. It is concluded that the nonsense-mutated eukaryotic expression vectors of fIX gene have been successfully constructed and can express in COS-7 cells, which provides the material basis for further researches on mechanism and treatment of FIX deficiency and the function defects caused by nonsense mutation.
Animals ; Base Sequence ; COS Cells ; Cercopithecus aethiops ; Cloning, Molecular ; Codon, Nonsense ; genetics ; Factor IX ; genetics ; Genetic Vectors ; Transfection