Apoptosis ; Humans ; Periodontitis ; pathology

Animals ; Biomarkers ; analysis ; metabolism ; Carcinoma, Hepatocellular ; diagnosis ; metabolism ; Chromatography, High Pressure Liquid ; methods ; Discriminant Analysis ; Fatty Liver ; diagnosis ; metabolism ; Humans ; Liver Failure ; diagnosis ; metabolism ; Liver Neoplasms ; diagnosis ; metabolism ; Mass Spectrometry ; methods ; Metabolomics

Actinomyces ; isolation & purification ; Actinomycosis ; microbiology ; Apicoectomy ; methods ; Candida albicans ; isolation & purification ; Candidiasis ; microbiology ; Enterococcus faecalis ; isolation & purification ; Foreign Bodies ; complications ; Gram-Positive Bacterial Infections ; microbiology ; Humans ; Microsurgery ; methods ; Periapical Periodontitis ; etiology ; microbiology ; surgery ; therapy ; Radicular Cyst ; complications ; Root Canal Filling Materials ; therapeutic use ; Root Canal Therapy ; methods

To study the effect of sodium aescinate in inducing human breast cancer MCF-7 cells apoptosis and its possible mechanism. MTT assay was used to detect the inhibitory effect of sodium aescinate on the proliferation of MCF-7 cells. The morphological changes were observed under inverted microscope. DAPI nuclear staining was used to detect the changes in cell nucleus. Annexin V-FITC/PI flow cytometry was adopted to test the apoptosis rate. Changes in apoptosis-related proteins (PARP, cleaved caspase-8 and pro-caspase-3), cell survival-associated signal molecules (AKT and ERK) and their common upstream kinase SRC was detected by Western blotting. The result showed that after different concentrations of sodium aescinate were used to treat breast cancer MCF-7 cells, they inhibited the proliferation of MCF-7 cells in a dose-dependent manner, induced cell apoptosis (typical morphological changes in nucleus, significant increase in cell apoptosis rate). The expressions of cleaved PARP and caspase-8 increased, while the expression of pro-caspase-3 decreased, which further verified sodium aescinate's effect in inducing cell apoptosis. Sodium aescinate significantly inhibited the phosphorylation of cell survival-related signal molecules (AKT, ERK) and down-regulate the activation of their common up-stream kinase SRC. The findings indicated that sodium aescinate can block signals transiting to downstream molecules AKT, ERK, inhibit the proliferation of breast cancer cell MCF-7 cell apoptosis and induced cell apoptosis by suppressing the activation of SRC.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Breast Neoplasms ; drug therapy ; enzymology ; genetics ; physiopathology ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; genetics ; metabolism ; Female ; Humans ; MCF-7 Cells ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Saponins ; pharmacology ; Signal Transduction ; drug effects ; Triterpenes ; pharmacology ; src-Family Kinases ; genetics ; metabolism

Objective To explore the clinical characteristics of cardiac involvement in children with mitochondriopathies.Methods The clinical data of 23 children with mitochondriopathies were reviewed.The changes of electrocardiography,echocardiography and heart enzymes were analyzed.Results In 15 cases of mitochondrial encephalomyopathy,lactic acidosis,and stroke-like episode(MELAS syndrome),electrocardiography was performed on 9 cases,6 of them showed abnormal electrocardiographic findings,including right bundle branch block,ST-T change,Wolff-Parkinson-White syndrome,et al.On echocardiographic examination in 9 MELAS syndrome ca-ses,only 1 case showed hypertrophy cardiomyopathy.Six cases had increased plasma creatine kinaseMB(CK-MB) mass and only one of 12 MELAS syndrome cases had increased cardiac troponin I(cTnI) level.In 8 cases of subacute necrotizing encephalomyopathy(Leigh syndrome),electrocardiography was performed on 5 cases,4 of them showed abnormal electrocardiographic findings,including sinus tachycardia,ST-T change and low voltage.Two cases showed normal electrocardiography.Three out of 6 cases with Leigh syndrome showed increased plasma CK-MB mass.The molecular genetic examinations were performed in 13 cases of MELAS syndrome and 6 cases of Leigh syndrome.The mitochondrial DNA nt 3243 A→G mutation was found in white blood cells of 9 MELAS syndrome cases,the mutation rate being 37%-60%.The mitochondrial DNA nt 8993 T→C mutation was found in white blood cells of 2 Leigh syndrome cases.Conclusion In children with mitochondriopathies,myocardiac involvement is comparatively common,and even cardiomyopathy can occur.

Objective To investigate the expression of CXCR4 in cultured human dental pulp cells (HDPC) in vitro and the corresponding ligand SDF-1α level of HDPC supernatants stimulated by lipopolysaccharide(LPS)and tumor necrosis factor-α(TNF-α),and to explore the role of SDF-1αon the proliferation and the migration of HDPC.Methods The expression of CXCR4 in HDPC was detected by immunocytochemistry technique and indirect immunofluorescence technique.The culture supernatants of HDPC were collected after HDPC had been simulated by LPS and TNF-α of difierent concentrations for 48h and then the SDF-1α level was assayed by quantitative sandwich ELISA.Meanwhile,the effects of recombinant human SDF-1α(rhSDF-1α)on the proliferation and the migration of HDPC at different concentrations were observed by MTT and Boyden Chamber Assay.Results CXCR4 was expressed in cytomembrane of HDPC and SDF-1α was secreted into their normal cell supematants with a concentration of(4513.55±962.92)ng/L. The secretion of SDF-1α was both significantly decreased by stimulation with LPS and TNF-α(P＜0.05).In addition,rhSDF-1α stimulated the HDPC proliferation at the concentrations of 50,100,200μg/L(P＜0.01)and increased the chemotactic migration of HDPC significantly after 9h's incubation with the concentrations of 50,100μg/L(P＜0.05).Conclusions SDF-1α accelerated the proliferation and the migration of HDPC which expressed CXCR4.SDF-1-CXCR4 axis may play a role in repair of pulp injury.

Aim To investigate the influence of down-regulating adenosine A1 receptor and adenosine A2 A receptor gene expression on proliferation and activation of acetaldehyde-induced hepatic stellate cell-T6 cells through siRNA. Methods Alcoholic liver fibrosis in vitro model was constructed by inducing HSC-T6 cells with acetaldehyde. siRNA targeting A1R and A2AR were designed and synthesized according to its mRNA. The siRNA was transfected into rat HSC-T6 cells by li-posome LipofectamineTM 2000. HSC cell proliferation was measured by MTT. The mRNA levels of A1R, A2AR, α-SMA, Collagen I in the supernatant of the cell culture were measured by Quantitative Real-Time PCR. The protein levels of A1R, A2AR, α-SMA, Collagen I were measured by Western blot. Results A1 R and A2 AR siRNA effectively inhibited the cell proliferation, and they also significantly decreased the levels of A1R, A2AR,α-SMA, Collagen I, suggesting that A1 R and A2 AR might be potential target genes in the alcoholic liver fibrosis. Conclusions Silencing A1 R or A2 AR by RNAi can significantly inhibit the HSC proliferation, A1R and A2AR may be potential therapeutic target genes for alcoholic liver fibrosis.

OBJECTIVETo analyze the expression of foreign gene in the filial generation of the transgenic plants on the base of the original transgenic tomatoes seeds carrying the gene encoding saliva-binding region (SBR) in PAc of Streptococcus mutans (S. mutans) gained.

METHODSThe tomatoes total DNA was extracted by CTAB methods, and the filial generation transgenic tomatoes carrying the gene encoding SBR in PAc of S. mutans were selected by PCR. The tomatoes total RNA was extracted by trizol and the transcription of the foreign gene was analyzed by RT-PCR. Protein was extracted from fruit tissue and the content of the total protein was determined by Bradford's methods G250. The expression of foreign protein was analyzed by Western blot and the lever of the foreign protein was analyzed by ELISA.

RESULTSThe fragment encoding SBR in S. mutans PAc gene integrated in the tomato genomic DNA and was expressed. The foreign protein lever was up to 1.2% of the total soluble protein in tomato fruit tissue.

CONCLUSIONThe foreign protein gene in the filial generation of the transgenic plants could express the foreign protein.

Lycopersicon esculentum ; Saliva ; Streptococcus mutans

OBJECTIVEThis study was carried out to investigate the effects of insulin-like growth factor-II (IGF-II) and basic fibroblast growth factor (bFGF) on osteoprotegerin (OPG) secretion of periodontal ligament cells (PDLCs).

METHODSHealthy human premolars extracted for orthodontic reasons from 12-14 years old donators were obtained, and periodontal tissues were collected and cultured to obtain PDL cells. Primary or first passage PDLCs were cloned by means of limited dilutions. PDLCs with osteoblastic phenotypes were characterized as follows: Alkaline phosphatase activity, collagen III production and bone-like nodules formation. IGF-II and bFGF were added into culture media and their effects on PDLCs proliferation and OPG secretion were observed. The OPG concentrations in cell culture supernatants were detected by sandwich ELISA. Living cell numbers were demonstrated by MTT test. The average levels of OPG secretion by a single cell were calculated by dividing OPG concentration with MTT-test result.

RESULTSBoth IGF-II and bFGF upregulated the mtt values (P < 0.05), but ICF-II downregulated the opg/mtt values (P < 0.05), whereas bFGF had no significant effect on opg/mtt values (P > 0.05).

CONCLUSIONIGF-II enhances the proliferation of PDL cells but prohibits OPG secretion. Although bFGF has the same effect on the proliferation of PDL cells, it has no effect on OPG secretion. Before cytokines were used to enhance periodontal regeneration, their effects on local bone balance should also be studied.

Adolescent ; Cells, Cultured ; Child ; Fibroblast Growth Factor 2 ; pharmacology ; Glycoproteins ; biosynthesis ; Humans ; Insulin-Like Growth Factor II ; pharmacology ; Osteoprotegerin ; Periodontal Ligament ; cytology ; drug effects ; metabolism ; Receptors, Cytoplasmic and Nuclear ; biosynthesis ; Receptors, Tumor Necrosis Factor ; biosynthesis

Objective To investigate the health effects on practitioners occupationally exposed to acrylonitrile and to provide scientific bases for the study of toxicological effects of acrylonitrile on human bodies. Methods 465 medical practitioners exposed to acrylonitrile and ( a control group of ) 488 medical practitioners unexposed to acrylonitrile were selected .Age, smoking habits , alcoholic habits and other relevant factors are well-considered in selecting these two groups and all of them had lived in Ningbo for at least three years .Blood samples were collected to measure the level of Alanine aminotransferase ( ALT) , aspartate aminotransferase ( AST ) , alkaline phosphatase ( ALP ) , pancreatic acyl transferase (GGT), total bilirubin (STB), blood urea (UREA), blood uric acid (UA), serum creatinine (SCr), total protein ( TP ) , albumin ( Alb ) , globulin ( Glb ) , ratio of Alb to Glb ( A/G ) , white blood cell ( WBC ) , red blood cell ( RBC ) , platelet ( PLT ) and hemoglobin ( Hb ) .These serum biochemical indices of the two groups were compared to find the differences thereof and to examine the effect of work years on various above-mentioned indices for medical practitioners exposed to acrylonitrile . Results The levels of ALT(t=-2.77,P=0.006), AST(t=-5.74,P<0.001), UREA(t=3.51,P<0.001), UA (t=-3.51,P<0.001)and SCr(t=-7.62,P<0.001)for medical practitioners exposed to acrylonitrile were all significantly higher than those for the control group; however, the levels of ALP(t=18.87,P<0.001), Alb(t=6.92,P<0.001), Glb(t=7.99,P<0.001), A/G(t=11.93,P<0.001), and WBC (t=4.48,P<0.001) were all lower than those for the control group , with the exposure time exhibiting positive correlations with ALT(r=0.564,P<0.001)and AST(r=0.493,P<0.001). Conclusion Acrylonitrile has certain health effects on the hepatorenal function as well as the routine blood test results of medical practitioners occupationally exposed to acrylonitrile .