Ac-Phe-Lys-PABC-DOX (PDOX) is a smart doxorubicin (DOX) prodrug designed to decrease toxicities while maintaining the potent anticancer effects of DOX. This study was aimed at elucidating the effectiveness and toxicities of DOX and PDOX in patient-derived MCF-7 breast cancer cells in vitro. The MCF-7 cells were exposed to both PDOX and DOX, and cytotoxicities, cell cycle and P53/P21 signaling alterations were studied. Abundant cathepsin B was found in the MCF-7 cells, and treatment with PDOX and DOX triggered dose- and time-dependent cytotoxicity and resulted in a significant reduction in cell viability. The IC50 of PDOX and DOX was 3.91 and 0.94 μmol/L, respectively. Both PDOX and DOX caused an up-regulation of the P53/P21-related signal pathway, and PDOX significantly increased expression of P53 and caspase 3, and arrested the cell cycle at the G1/G2 phase. As compared with DOX, PDOX reduced toxicities, and it may have different action mechanisms on breast cancer cells.

ObjectiveTo observe the effects of bone marrow stromal cells (BMSCs) on vessel endothelial cells proliferation and microvessel formation in vitro.MethodsBMSCs and brain vessel endothelial cells were separated from adult and divided into co-culture group of BMSCs and endothelial cells, medium group of BMSCs, comparison group. Endothelial cells proliferation and microvessel formation were observed. ResultsEndothelial cells were promoted to proliferate and formate the microvessel in medium group and co-culture group. And the effect was prominence in co-culture group.ConclusionBMSCs can promote the proliferation and microvessel formation of endothelial cells.

Objective To examine the expression of recombinant cytolethal distending toxin(CDT)produced by Actinobacillus actinomycetemcomitans(Aa).Methods CDT encoding gene cdtABC was amplified by PCR.Through TA clone and restriction endonuclease digestion,gene cdtABC and vector pQE60 were ligated to form pQE60-cdtABC expression system which transformed into competent cells.Protein expression was induced by IPTG and examined by SDS-PAGE and Western-blotting.Results Random colony PCR of pQE60-cdtABC transformed cells demonstrated that all strains contained cdtABC gene.The DNA sequence was blast with cdtABC gene from GenBank and 99%homology was obtained.SDS-PAGE and Western-blotting confirmed that recombinant CDT was obtained.Conclusions CDT protein expression system was reconstructed and recombinant protein was obtained.

OBJECTIVETo explore the role of Xuebijing Injection (XBJI) in inhibiting inflammatory factors associated with anoxia/reoxygenation myocardial inflammatory response of rats.

METHODSTotally 36 healthy male Sprague-Dawley rats, 280 ± 30 g were randomly divided into six groups, i.e., the normal control group (N group), the balanced perfusion group (BP group),the model group (M group),the low dose XBJI group (XBJI(L) group), the middle dose XBJI group (XBJI(M) group),and the high dose XBJI group (XBJI(H) group), 6 in each group. The myocardial anoxia/reoxygenation rat model was established by Langendorff isolated heart perfusion. The concentration of TNF-α in the myocardial tissue was detected by ELISA. The expression of nuclear factor kappa B p65 (NF-κB p65) protein and Toll like receptor 4 (TLR4) protein were detected using Western blot. The expression of NF-κB p65 mRNA and TLR4 mRNA was detected by RT-PCR. Ultrastructural changes of anoxia-reoxygenation rats' heart muscle were observed under transmission electron microscope.

RESULTSCompared with the M group,the TNF-α concentration, expression levels of NF-κB p65 protein and mRNA, TLR4 protein and mRNA decreased to various degrees in the XBJI(L) group, the XBJI(M) group, and the XBJI(H) group. The TNF-α expression level decreased most significantly in the XBJI(L), group (P < 0.01), while other indices decreased most obviously in the XBJI(M) group (P < 0.01, P < 0.05). Expression levels of NF-κB p65 and TLR4 protein were obviously lower in the XBJI(M) group than in the XBJI(L) group (P < 0.05). There was no statistical difference in other indices among the three XBJI groups (P > 0.05). Myocardial fibers were loose and broken with disappearance of transverse striation, and mitochondrial cristae was dissolved and severely damaged in the M group. The aforesaid condition was improved after treated by XBJI, with the most obvious effect obtained in the XBJI(M) group.

CONCLUSIONSDifferent doses of XBJI could attenuate inflammatory reactions after myocardial anoxia/reoxygenation rats' heart muscle through inhibiting TLR4-NF-κB-TNF-α signal transduction pathway. The best effect could be obtained by 4 mL/100 mL XBJI.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Hypoxia ; Male ; Myocardium ; metabolism ; Myocytes, Cardiac ; NF-kappa B ; metabolism ; Oxygen ; metabolism ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Toll-Like Receptor 4 ; metabolism ; Transcription Factor RelA ; Tumor Necrosis Factor-alpha ; metabolism

Objective To evaluate the possible interactions among different impact factors possibly affecting the treatment efficacy of 131I in Graves′ disease (GD). Methods Six hundred and thirty two GD patients that had been treated by 131I, with or without antithyroid drugs (ATD), were included in this study. The impact factors were pre-defined as age (x1), sex (x2), mass of thyroid (x3), course of disease (x4), initial symptom (x5), condition of disease (x6), ATD treatment duration (x7), effective half life time (x8), maximum 131I uptake rate (x9), total dose of 131I (x10), dose of 131I per gram of thyroid (x11), TRAb (x12), TSI (x13), TgAb (x14), and thyroid microsomal antibody(TMAb) level(x15). Interactions among different impact factors were studied by t-test, χ2 test and multi-variant logistic regression. Results Age, mass of thyroid, ATD treatment duration, maximum 131I uptake rate, dose of 131I per gram of thyroid tissue and TSI level were identified as independent impact factors affecting the 131I treatment efficacy on GD (χ2=6.908, t=-4.063, χ2=13.558, t=-2.553, t=4.528, χ2=9.716, all P<0.05) by uni-variant and multi-variate analyses. Loglinear and general linear model analyses showed that there existed multiple multiplicative and additive interactions among the factors of age, mass of thyroid, ATD treatment duration and maximum 131I uptake rate (likelihood χ2=8.176, P>0.05; F=2.928, 1.992, 2.629, 2.215, all P<0.05), which indicated that the treatment efficacy with co-existing multiple factors was not equal to simple summation of single factors. Conclusions The interactions among multiple factors can cause indi-rect effect on 131I treatment, which might guide the prescription of 131I dosage for GD treatment.

OBJECTIVETo evaluate the effect of short-term intensive therapy on blood glucose control, BETA-cell function, and blood lipid levels in newly diagnosed type 2 diabetic patients.

METHODSOut-patients with newly diagnosed type 2 diabetic patients were enrolled for intensive treatment with sulfonylureas and metformin for 12 weeks, and the therapeutic effect was evaluated.

RESULTSAfter the intensive treatment, FPG, 2 hPG, and HbA1c decreased significantly (P<0.01); HOMA-IR decreased and HOMA-B increased significantly (P<0.01), and TG, CHOL, LDL decreased significantly (P<0.01) after the treatment.

CONCLUSIONShort-term intensive treatment with glimepiride combined with metformin is safe and effective in newly diagnosed type 2 diabetic patients with HbA1c>9%.

Diabetes Mellitus, Type 2 ; drug therapy ; Drug Therapy, Combination ; Female ; Humans ; Male ; Metformin ; therapeutic use ; Sulfonylurea Compounds ; therapeutic use ; Treatment Outcome

Ac-Phe-Lys-PABC-DOX (PDOX) is a smart doxorubicin (DOX) prodrug designed to decrease toxicities while maintaining the potent anticancer effects of DOX. This study was aimed at elucidating the effectiveness and toxicities of DOX and PDOX in patient-derived MCF-7 breast cancer cells in vitro. The MCF-7 cells were exposed to both PDOX and DOX, and cytotoxicities, cell cycle and P53/P21 signaling alterations were studied. Abundant cathepsin B was found in the MCF-7 cells, and treatment with PDOX and DOX triggered dose- and time-dependent cytotoxicity and resulted in a significant reduction in cell viability. The IC50 of PDOX and DOX was 3.91 and 0.94 μmol/L, respectively. Both PDOX and DOX caused an up-regulation of the P53/P21-related signal pathway, and PDOX significantly increased expression of P53 and caspase 3, and arrested the cell cycle at the G1/G2 phase. As compared with DOX, PDOX reduced toxicities, and it may have different action mechanisms on breast cancer cells.
Antibiotics, Antineoplastic ; pharmacology ; Breast Neoplasms ; drug therapy ; metabolism ; pathology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cyclin-Dependent Kinase Inhibitor p21 ; biosynthesis ; Doxorubicin ; analogs & derivatives ; pharmacology ; Drug Screening Assays, Antitumor ; methods ; Female ; G1 Phase ; drug effects ; G2 Phase ; drug effects ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Oligopeptides ; pharmacology ; Signal Transduction ; drug effects ; Tumor Suppressor Protein p53 ; biosynthesis

OBJECTIVETo compare the number of the Tannerella forsythensis (T. forsythensis), total bacteria, and proportion of T. forsythensis in subgingival specimens in diseased sites of chronic periodontitis patients and in healthy sites of periodontally healthy subjects, and clarify the relationship between bacterial load and periodontal status.

METHODSSubgingival plaque samples from 61 chronic periodontitis patients and 12 healthy controls (positive for T. forsythensis by conventional PCR) were analyzed with TaqMan real-time polymerase chain reaction assay for T. forsythensis and total bacteria. Quantification was performed with species-specific primer/probe, universal primer/ probe and serial dilution of plasmid standards.

RESULTSNumbers of T. forsythensis and total bacteria(P<0.001) , the proportion of T. forsythensis in subgingival specimens (P<0.05) were significant higher in diseased sites of chronic periodontitis patients than in healthy sites of healthy subjects. In addition, a significant correlation was found between the number of bacteria and various probing depth (P<0.001). There was no significantly difference between the proportion of T. forsythensis in subgingival plaque and probing depth.

CONCLUSIONNumber of T. forsythensis are closely associated with periodontal status, and demonstrate the broad potential of real-time polymerase chain reaction application on periodontology.

Adult ; Bacteroides ; Chronic Periodontitis ; Dental Plaque ; Female ; Humans ; Male ; Middle Aged ; Periodontitis ; Polymerase Chain Reaction ; Porphyromonas gingivalis

OBJECTIVETo detect the distribution of fimA genotype of P. gingivalis in periodontally healthy adults and chronic periodontitis patients, and to investigate the relationship between the prevalence of fimA genotype of P. gingivalis and periodontal health status.

METHODSSubgingival plaque samples were collected from 136 periodontally healthy adults and 115 chronic periodontitis patients. The occurrence of P. gingivalis was determined by P. gingivalis 16S rRNA PCR. Distribution of fimA genotype was assessed in P. gingivalis positive samples by PCR using primers pairs homologous to the different fimA genes.

RESULTSP. gingivalis was detected in 22.1% of the healthy subjects and 81.7% of chronic periodontitis patients. A single fimA genotype was detected in most subgingival plaque samples. In P. gingivalis-positive healthy adults, the most prevalent fimA genotype of P. gingivalis was type I fimA. In contrast, a majority of chronic periodontitis patients carried type II fimA, followed by IV fimA and I b fimA. The univariate analysis illustrated that chronic periodontitis was associated with occurrences of type I fimA (OR = 0.97), I b (OR =13.26), II (OR = 36.62), III (OR = 4.57), IV (OR = 22.86), and V (OR = 1.19).

CONCLUSIONII fimA genotype of P. gingivalis followed by IV and I b were an important virulence factor that may account for the pathogenesis of chronic periodontitis, suggesting an increased pathogenic potential of these types.

Adult ; Chronic Periodontitis ; Dental Plaque ; Female ; Fimbriae Proteins ; Genotype ; Health Status ; Humans ; Male ; Periodontitis ; Polymerase Chain Reaction ; Porphyromonas gingivalis ; Prevalence ; RNA, Ribosomal, 16S

OBJECTIVEThe aim of this study was to investigate the prevalence of T. forsythensis in Chinese chronic periodontitis patients, and correlate it with clinical parameters.

METHODS108 chronic periodontitis patients were selected according to clinical criteria. In each patient, subgingival plaque samples were collected from two of the periodontitis involved molar sites with the deepest periodontal pocket depth(diseased sites) and one healthy gingival sulcus (healthy sites). T. forsythensis was detected by 16S rRNA polymerase chain reaction. Clinical assessments including probing depth, clinical attachment loss and bleeding on probing, were made at sampling sites.

RESULTSThe prevalence of Tforsythensis in diseased sites and healthy sites was 59.72% and 3.70% respectively (P < 0.001). The prevalence of T. forsythensis was significantly higher in deeper pockets and more serious clinical attachment loss, Spearman rank correlation coefficient was 0.48 and 0.51 respectively. The prevalence of T. forsythensis in BOP (+)sites was 61.31%, and in BOP(-) sites was 8.80% (P < 0.001).

CONCLUSIONThe prevalence of T. forsythensis was significantly correlated with clinical parameters, suggesting that T. forsythensis is closely related to chronic periodontitis in the Chinese population.

Adult ; Bacteria ; Chronic Periodontitis ; Dental Plaque ; Female ; Gingiva ; Humans ; Male ; Middle Aged ; Periodontal Pocket ; Periodontitis ; Polymerase Chain Reaction ; Prevalence ; RNA, Ribosomal, 16S