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.

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

OBJECTIVETo examine the expression of recombinant cytolethal distending toxin (CDT) produced by Actinobacillus actinomycetemcomitans (Aa).

METHODSCDT 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.

RESULTSRandom 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.

CONCLUSIONSCDT protein expression system was reconstructed and recombinant protein was obtained. Actinobacillus actinomycetemcomitans;

Aggregatibacter actinomycetemcomitans ; genetics ; metabolism ; Bacterial Toxins ; genetics ; metabolism ; Genetic Vectors ; Recombinant Proteins ; genetics ; 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 investigate the effects of C-reactive protein (CRP) on monocytes chemotaxis ability in vitro.

METHODSTranswell chemotaxis assay was used to evaluate the changes of chemotactic ability of THP-1 monocytes in each group treated with CRP in different concentration.

RESULTSCRP increased the number of attracted monocytes in response to MCP-1 (monocyte chemoattractant protein-1). When treated with CRP concentration at 2 microg x mL(-1), the number of chemotactic monocytes increased (P < 0.05). The number of attracted monocytes increased as CRP concentration was elevated (P < 0.05).

CONCLUSIONCRP can increase chemotactic ability of THP-1 monocytes in concentration dependent manner.

C-Reactive Protein ; Chemokine CCL2 ; Chemotaxis ; Humans ; In Vitro Techniques ; Monocytes

Toxicity of different processed was evaluated Polygoni Multiflori Radix by determining the hepatotoxic potency for selecting processing technology. Process Polygoni Multiflori Radix using high pressure steamed, Black Bean high pressure steamed, atmospheric steamed for different time. Using normal human hepatocytes (L02) as evaluation model, hepatotoxic potency as index to evaluate hepatotoxic potency of different processed Polygoni Multiflori Radix. Analysis chemical composition of some processed products by UPLC-MS. Hepatotoxic bioassay method cloud evaluate the toxicity of different Polygoni Multiflori Radix samples. Different processing methods can reduce the toxicity of Polygoni Multiflori Radix, high pressure steamed three hours attenuated was better. Different processing methods have different effects on chemical constituents of Polygoni Multiflori Radix. Comparing with crude sample, the contents of gallic acid, 2,3,5,4-tetrahydroxyl diphenylethylene-2-O-glucoside, emodin-8-O-beta glucoside and emodin were decreased in processed products with 3 kinds of different methods. The change trend of 2,3,5,4-tetrahydroxyl diphenylethylene-2-O-glucoside content was similar with hepatotoxic potency. Different processing methods can reduce the toxicity of Polygoni Multiflori Radix. Processing methods and time attenuated obvious impact on toxicity. Recommended further research on the attehuated standard control of Polygoni Multiflori Radix concocted.
Biological Assay ; Cell Line ; Chemistry, Pharmaceutical ; methods ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; toxicity ; Fallopia multiflora ; chemistry ; toxicity ; Hepatocytes ; drug effects ; Humans ; Plant Roots ; chemistry ; toxicity

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

OBJECTIVETo investigate the prevalence of H. actinomycetemcomitans in Chinese chronic periodontitis (CP) patients and periodontally healthy adults.

METHODS116 chronic periodontitis patients and 111 periodontally healthy adults were included. In each CP patient, subgingival plaque samples were collected from two sites of different molars with the greatest probing depth (PD) and one periodontally healthy site (PD < or =3 mm). The samples of periodontally healthy adults were obtained from the mesio-buccal site of one first upper molar. Bacteria DNA were extracted for detection of H. actinomycetemcomitans by 16S rRNA PCR.

RESULTSThe prevalence for H. actinomycetemcomitans of diseased sites (33.62%) was significantly higher than that of healthy sites from CP patients (0.86%) and the periodontally sites (0.90%) (P < 0.01). No significant difference was observed between male and female CP patients (P > 0.05). A decreasing trend of H. actinomycetemcomitans was observed as the age increased. And the pocket depth and clinical attachment losswas associated with the occurrence of H. actinomycetemcomitans in a positive mode. And H. actinomycetemcomitans was more often detected in the bleeding sites on probing.

CONCLUSIONH. actinomycetemcomitans was more frequently detected in periodontitis sites than periodontally healthy sites. For CP patients, a higher prevalence was associated with the seriously involved sites than those moderate and mild implicated sites. H. actinomycetemcomitans is considered to be the one of the periopathogens involved in the etiology of chronic periodontitis.

Adult ; Aggregatibacter actinomycetemcomitans ; Chronic Periodontitis ; DNA, Bacterial ; Dental Plaque ; Female ; Healthy Volunteers ; Humans ; Male ; Periodontitis ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S