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 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

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

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 establish PEG10 transgenic mice model and study the effect of PEG10 transgene on tumor growth and metastasis in mice.

METHODSThe linearized expression element of pALB-PEG10, which contained mouse albumin promoter, structural gene of PEG10, and polyaenylation signal sequence, was microinjected into 3741 KM mouse fertilized ova. The manipulated embryos were then transplanted into the oviducts of 94 pseudopregnant recipient mice. All the newborn mice were screened by PCR to detect genomic DNA in tail tissue, then PEG10 mRNA and protein expression were detected by RT-PCR and western blot, respectively in the positive mice. Hepatoma cell H22 was subcutaneously inoculated into the right armpit of wild type mice and No.17, No.33 transgenic mice. Tumor size was measured every week. Mice were sacrificed on day 12 and then the tumors were exercised and weighted. Tumors and livers were fixed in formaldehyde and sectioned. The sections were stained with hematoxylin/eosin and examined under microscope. The expression of PEG10 protein was detected with immunohistochemistry method.

RESULTSAmong the 43 off-springs, 3 were positive for tail tissue PEG10 gene examination, PEG10 was successfully expressed in the liver of the randomly selected transgenic mouse. H22 tumor grew faster in all the transgenic mice than in wild type mice. The average size and weight of tumors between the transgenic mice and wild type mice were significantly different (P < 0.05). Most tumors in the transgenic mice invaded the surrounding tissues and showed liver metastasis, PEG10 protein was expressed in liver. In contrast, nearly all the tumors in wild type mice were capsulized and PEG10 was not expressed in liver.

CONCLUSIONOur results showed that the PEG10 gene could be expressed in the liver of the transgenic mice. PEG10 promotes growth, invasion, and metastasis of transplanted H22 tumors in mice.

Animals ; Cell Line, Tumor ; Disease Models, Animal ; Genetic Vectors ; Humans ; Liver ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Mice ; Mice, Transgenic ; genetics ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Transplantation ; Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transgenes ; Xenograft Model Antitumor Assays

Post-translational degradation of protein plays an important role in cell life. We employed chimeric molecules (dihydrotestosterone-based proteolysis-targeting chimeric molecule [DHT-PROTAC]) to facilitate androgen receptor (AR) degradation via the ubiquitin-proteasome pathway (UPP) and to investigate the role of AR in cell proliferation and viability in androgen-sensitive prostate cancer cells. Western blot analysis and immunohistochemistry were applied to analyse AR levels in LNCaP cells after DHT-PROTAC treatment. Cell counting and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) cell viability assay were used to evaluate cell proliferation and viability after AR elimination in both LNCaP and PC-3 cells. AR was tagged for elimination via the UPP by DHT-PROTAC, and this could be blocked by proteasome inhibitors. Degradation of AR depended on DHT-PROTAC concentration, and either DHT or an ALAPYIP-(arg)(8) peptide could compete with DHT-PROTAC. Inhibition of cell proliferation and decreased viability were observed in LNCaP cells, but not in PC-3 or 786-O cells after DHT-PROTAC treatment. These data indicate that AR elimination is facilitated via the UPP by DHT-PROTAC, and that the growth of LNCaP cells is repressed after AR degradation.
Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Dihydrotestosterone ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; Male ; Prostatic Neoplasms ; drug therapy ; metabolism ; pathology ; Proteasome Endopeptidase Complex ; metabolism ; Receptors, Androgen ; metabolism ; Recombinant Fusion Proteins ; pharmacology ; therapeutic use ; Signal Transduction ; drug effects ; Ubiquitin ; metabolism

OBJECTIVETo apply multiplex polymerase chain reaction (MPCR) assay and sequencing in study of the carrying status of four pathogenicity-related genes of Vibrio cholerae (V.cholerae) and the variation of ctxA.

METHODSPrimers targeting cholera toxin sub-unit A gene (ctxA), toxin-coregulated pilus gene (tcpA), accessory cholera enterotoxin gene (ace), zonula occludens toxin gene (zot) were designed and the MPCR method was applied to detect the pathogenicity-related genes of 276 strains of V.cholerae isolates. The amplified fragments of ctxA gene were sequenced and the genetic homology of the amplified fragments of ctxA was analyzed.

RESULTSOf the 276 strains of V.cholerae, 93.9% strains from human sources belong to the pathogenicity-related genes type A (ctxA(+)tcpA(+)ace(+)zot(+) type) and 6.1% belong to pathogenicity-related genes type C (ctxA(-)tcpA(-)ace(-)zot(-) type). Type A strains from clinical sources were isolated from patients with mild to severe symptom and carriers, among which 68.5% were isolated from patients with mild symptom and 21.9% from carriers. All 63.6% of type C strains from clinical sources were isolated from patients with mild symptom and 36.4% from carriers. The proportion of type C strains that caused mild symptom was higher than that of type A strains. Of the 78 strains isolated from the environment, 9.0% strains belong to pathogenicity-related type A and 35.9% belong to the pathogenicity-related genes type B (ctxA(-)tcpA(-)ace(+)zot(+) type), while 55.1% belong to pathogenicity-related genes type C. The sequencing results showed little genetic variation among the amplified fragments for ctxA.

CONCLUSIONMPCR disclosed the polymorphic status of pathogenicity-related gene patterns in V.cholerae isolates of Guangzhou, providing effective means for further study on evolution of pathogenicity-related genes among V.cholerae isolates from human and environmental sources. This study also offers significant guidance for effective prevention, control and warning against cholera epidemic in local area.

China ; Cholera Toxin ; genetics ; DNA, Bacterial ; Genes, Bacterial ; genetics ; Genotype ; Humans ; Polymerase Chain Reaction ; Sequence Analysis ; Vibrio cholerae ; classification ; genetics ; isolation & purification

OBJECTIVETo investigate the ability of the pericardium meridian (PM) to mitigate or enhance the cardiotoxic effects of aconitine injected at specific acupoint and non-acupoint sites in rabbits.

METHODSThis study consisted of 3 experiments that were designed to test the effects of injection of 30 μg/kg of aconitine at acupoints on the PM (Test 1), at non-acupoint sites on the PM (Test 2), and at acupoints on other meridians and non-meridian sites (Test 3). In Test 1, 24 rabbits were randomly assigned to receive injections at Quze (PC3), Tianquan (PC2), or intramuscularly. In Test 2, 24 rabbits were randomly assigned to receive injections of aconitine at non-acupoint I, non-acupoint II, or intramuscularly. In Test 3, 48 rabbits were randomly assigned to receive injections at Neiguan (PC6), Sanyinjiao (SP6), Yangjiao (GB35), a non-meridian and non-acupoint site (NMNA), intravenously, and intramuscularly. Electrocardiographs of the rabbits were performed before, during and after injection to determine the incidence of arrhythmia, latency of ventricular rhythm, and recovery rate after aconitine injection. The recovery time index and extent of arrhythmia scores were calculated.

RESULTSIn all groups the incidence of arrhythmia was 100%, and the latency of ventricular rhythm was less than 30 min. In Tests 1 and 2, the recovery rates of the Quze and non-acupoint II groups were significantly higher than those of the muscular group (P < 0.05). In Test 3, the recovery time index and extent of arrhythmia scores of the Neiguan group were low. There were no significant differences between the other acupoint groups, or the NMNA group, when compared with the group receiving aconitine intramuscularly.

CONCLUSIONSAcupoints or non-acupoints along the PM could reduce the severity of the arrhythmia induced by aconitine in healthy rabbits. Meridians play an important role in protecting body functions.

Aconitine ; adverse effects ; pharmacology ; Acupuncture Points ; Acupuncture Therapy ; methods ; Analysis of Variance ; Animals ; Arrhythmias, Cardiac ; chemically induced ; diagnosis ; Disease Models, Animal ; Electrocardiography ; Male ; Meridians ; Pericardium ; drug effects ; Rabbits ; Random Allocation