OBJECTIVETo detect and compare the activity and intensity of gingipain K (Kgp)-caspase like subdomain in culture medium and cell extract of Porphyromonas gingivalis (Pg) isolates in puberty gingivitis and to reveal the possible association of Kgp with puberty gingivitis.

METHODSThirty-six children of 14 to 17 years old were enrolled in this study. Clinical parameters including gingival index (GI), sulcus bleeding index (SBI) and probing depth (PD) were evaluated. Subgingival plaque samples were collected and Pg isolates were obtained. 16S rRNA PCR was used to confirm Pg clinical isolates. Bacteria were grown in batches of BHI base and harvested at the end of log-phase growth. Culture fractions (culture medium and cell extract) of 10 Pg isolates were performed with SDS-PAGE and Western blot technique using primary antibody against specific Kgp-caspase like subdomain. Activity of Kgp in both samples was detected as well. The data were statistically analyzed using SPSS 11.5 software. The relationship between the Kgp intensity/activity of Kgp and the clinical parameters was statistically analyzed using Spearman correlation coefficient.

RESULTSThere was positive correlation between the intensity/activity of Kgp and the clinical parameters (P < 0.05).

CONCLUSIONSThe Kgp in clinical isolates of Pg from puberty gingivitis is in complicated forms. Caspase-like molecules with low molecular weight may exist as intracellular functional protein molecules which can affect the interaction between Pg and host. Kgp was contributes in certain degree to the pathogenesis of puberty gingivitis.

Adhesins, Bacterial ; genetics ; metabolism ; Adolescent ; Cysteine Endopeptidases ; genetics ; metabolism ; Dental Plaque ; microbiology ; Female ; Gingivitis ; enzymology ; microbiology ; Humans ; Male ; Porphyromonas gingivalis ; genetics ; isolation & purification ; metabolism

OBJECTIVETo investigate DNA aneuploid and P16 expression in biopsy specimens from lung cancer, and to study genetic instability and the application of flow cytometry in lung cancer pernicious degree diagnosis.

METHODSBlood cells and cancer cells in biopsy specimens were marked simultaneously with anti-CD45 and anti-P16 fluorescent antibody, and the ratio of CD45+ P16+ cells and CD4- P16+ cells was compared. DNA content in biopsy specimens from lung cancer was detected by flow cytometry.

RESULTSAmong the 74 cases of lung cancer, there are 46 cases of DNA aneuploid (62.2%). Thirty-seven cases of lung cancer expressed P16 lowly (50%). Twelve cases of lung cancer only expressed P16 lowly (16.22%), 21 cases of lung cancer only expressed DNA aneuploid (28.38%), and 25 cases not only expressed P16 lowly but also expressed DNA aneuploid (33.78%). Indexes of malign degree, such as P16 low expression or DNA aneuploid could be detected in 58 cases among the 74 cases (78.38%) by flow cytometry.

CONCLUSIONP16 low expression and DNA aneuploid are the indexes of lung cancer malign degree, and flow cytometry can be used to study genetic instability and evaluate biopsy specimens from lung cancer.

Aneuploidy ; Animals ; Biopsy ; Chromosomal Instability ; genetics ; DNA ; genetics ; Female ; Flow Cytometry ; Gene Dosage ; Gene Expression Regulation, Neoplastic ; Genes, p16 ; Humans ; Leukocyte Common Antigens ; genetics ; Lung Neoplasms ; diagnosis ; genetics ; pathology ; Male ; Mice ; Middle Aged

OBJECTIVETo investigate the cytotoxic mechanism of cadmium (Cd) on cerebral cortical neurons.

METHODSThe primary cultures of rat cerebral cortical neurons were treated with different concentrations of cadmium acetate (0, 5, 10, and 20 micromol/L), and then the cell viability, apoptosis, ultrastructure, intracellular [Ca2+], and reactive oxygen species (ROS) levels, mitochondrial membrane potential (delta psi), activities of catalase (CAT) and superoxide dismutase (SOD) were measured.

RESULTSA progressive loss in cell viability and an increased number of apoptotic cells were observed. In addition, Cd-induced apoptotic morphological changes in cerebral cortical neurons were also demonstrated by Hoechst 33258 staining. Meanwhile, ultrastructural changes were distortion of mitochondrial cristae and an unusual arrangement. Simultaneously, elevation of intracellular [Ca2+]i and ROS levels, depletion of Delta Psi were revealed in a dose-dependent manner during the exposure. Moreover, CAT and SOD activities in the living cells increased significantly.

CONCLUSIONExposure of cortical neurons to different doses of Cd led to cellular death, mediated by an apoptotic mechanism, and the apoptotic death induced by oxidative stress may be a potential reason. And the disorder of intracellular homeostasis caused by oxidative stress and mitochondrial dysfunction may be a trigger for apoptosis in cortical neurons.

Animals ; Apoptosis ; drug effects ; Cadmium ; toxicity ; Cerebral Cortex ; cytology ; drug effects ; metabolism ; In Vitro Techniques ; Neurons ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism

OBJECTIVETo examine the role of Polo-like kinase 1(PLK1) in the migration and invasiveness of human colorectal cancer cells.

METHODSNine colorectal cancer cell lines were cultured. Cell lines with the highest level of PLK1 expression was selected by PCR and Western blot. Three siRNA oligo segments targeting PLK1 were designed and selected cell lines transfected. Successful transfection was confirmed using real-time PCR and Western blot. Changes in migration and invasiveness of the selected cell line were evaluated by Transwell test.

RESULTSColorectal cancer cell line SW1116 was selected with the highest expression of PLK1 at both mRNA level and protein level. The expression of PLK1 in SW1116 was reduced by the three siRNA oligo segments to varying degrees, and the No.1 siRNA oligo segment was the most efficient. In migration test, the number of cells crossing through chambers in PLK1-siRNA group was 44 ± 14, which was lower than that in the negative control group (242 ± 40) and in blank control group(240 ± 38). In invasion test, the number of cells crossing through chambers in PLK1-siRNA group was 62 ± 3, which was lower than that in negative control group (207 ± 12) and in blank control group (211 ± 15). These differences were statistically significant(P<0.01).

CONCLUSIONPLK1 silencing by siRNA may inhibit the migration and invasiveness of colorectal cancer cells, suggesting that PLK1 might play an important role in the infiltration and metastasis of colorectal cancer.

Cell Cycle Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Cell Movement ; Colorectal Neoplasms ; metabolism ; pathology ; Humans ; Neoplasm Invasiveness ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Transfection

A rice pse(t) (premature senescence, tentatively) mutant line, was isolated from 4,500 independent T-DNA inserted transgenic lines. The symptoms of premature senescence appeared more severely than those of the control plants (Zhonghua 11, japonica) at the last development stage. To characterize the mutant and provide basic information on the candidate genes by mapping to a physical region of 220-kb, experiments were carried out in two phytotrons under controlled temperature of 24 degrees C and 28 degrees C, respectively. The content of chlorophyll, soluble protein and MDA (malondialdehyde), net photosynthesis, the antioxidant enzyme activities of SOD (superoxide dismuase) (EC 1.15.1.1) and POD (peroxidase) (EC 1.11.1.7) and the peptidase activities of leaves were measured from top to bottom according to the leaf positions at the flowering stage. Compared with the control plant, the mutant showed the following characteristics: (1) Higher net photosynthesis rate (P(n)) appeared in the 1st and 2nd leaves, contents of chlorophyll and soluble protein were also higher in the 1st leaf; (2) The activities of SOD, POD and peptidase were higher according to the leaf position from top to bottom; (3) The symptom of premature senescence was accelerated in the mutant at 28 degrees C treatment. The MDA content and the SOD and POD activities between the 24 degrees C and 28 degrees C treatment mutants were not significantly different. Content of chlorophyll and soluble protein of leaves mutant decreased rapidly at 28 degrees C treatment. The results show that pse(t) is sensitive to high temperature. The probable function of PSE(T) is discussed.
Aging ; physiology ; Antioxidants ; metabolism ; Apoptosis ; physiology ; Mutation ; Oryza ; classification ; genetics ; growth & development ; Photosynthesis ; physiology ; Plant Leaves ; classification ; genetics ; growth & development ; Plant Proteins ; genetics ; metabolism

OBJECTIVETo probe into the effects of familial factors on injury-related behaviors in children.

METHODSInjury-related behaviors and familial factors of 6884 children were investigated with Family Questionnaire and Child Behavior Checklist. Multi-nominal logistic regression analysis was performed.

RESULTSThere were 1670 (24.26%) children having serious injury-related behaviors and 3683 (53.50%) children having moderate injury-related behaviors. Factors contributing to children's injury-related behaviors would include punishment or indifference as well as the mode of parents' education; reintegral type of family; the level of parents' cognition on injuries; unfit location of medicine at home and careless attitudes of parents.

CONCLUSIONThere was close relationship between children's injury-related behaviors and familial factors. To avoid injury-related behaviors and to prevent injury occurrence, the importance of familial factors must be stressed.

Child ; Child Behavior ; psychology ; Family ; psychology ; Female ; Humans ; Logistic Models ; Male ; Surveys and Questionnaires ; Wounds and Injuries ; etiology ; prevention & control ; psychology

Objective To evaluate the laparoscopic management in diagnosing and treating small intestinal stromal tumors.MethodsSeventeen patients with small intestinal stromal tumors,who underwent laparoscopic diagnosis and surgical procedures,were retrospectively analyzed with the size of tumor,operative time,blood loss,length of incision,time for passage of flatus,post-operative hospital stay,operative complications and result of follow-up.Results All the 17 patients were diagnosed and treated by laparoscopic partial intestinal resections.The mean diameter of tumors was(3.6?1.3)cm,operative time(62.1?25.7)min,blood loss(17.1?15.2)mL,length of incision(3.4?1.1)cm,time for passage of flatus(2.2?1.2)d and post-operative hospital stay(7.8?2.0)d.As for the complications,one(5.9%)anastomosis bleeding was found and was treated by non-surgical procedure.After follow-up for 2-34 months,no recurrent tumor was found.Conclusion Laparoscopic exploration is useful to diagnose small intestinal stromal tumor accurately,and laparoscopic partial intestinal resection is a safe,effective and less invasive procedure for small intestinal stromal tumors.

Objective To evaluate the clinical application of laparoscopic local resection for gastric tumors.Methods Twenty-three patients with gastric tumors who were performed laparoscopic gastric local resection were retrospectively analyzed with the size of tumor,location of tumor,operative time,blood loss during the operation,time for passage of flatus,post-operative hospital stay,operative complications,post-operative pathological findings and result of follow-up.Results Twenty-three patients were successfully performed laparoscopic local resection,including 8 laparoscopic wedge resection(LWR)and 15 intragastric mucosal resections(IGMR),with no conversion to open surgery.The mean size of gastric tumor was(2.8?1.3)cm,the mean operative time was(82.2?35.5)min,the mean blood loss was(26.5?15.3)mL,the length of incision was(3.1?1.1)cm,the time for passage of flatus was(2.1?0.9)d,and the mean post-operative hospital stay was(7.8?2.0)d.Two patients(8.7%)were found with postoperative gastric mucosal blee-ding and were recovered well through non-operative treatment.The median time of follow-up was 12 months(2-45 months),and no recurrent tumor was observed.Conclusion Laparoscopic local resection is a feasible,safe,effective and less invasive procedure for gastric tumors.

Objective To investigate the value of endoscopy combined with laparoscopy in the treatment of colorectal polyps and polyp canceration.Methods Different combinations of endoscopic and laparoscopic procedures were employed and the clinical efficacies were compared.Results From January 2004 to September 2006,46 cases with colorectal polyp were treated with endoscopy combined with laparoscopy.Among them,5 cases(10.87%)underwent laparoscopic-assisted endoscopic polypectomy,30(65.22%)endoscopic-assisted laparoscopic resection,6(13.04%)synchronously endoscopic and laparoscopic resection.Five cases were performed further operation after endoscopic polypectomy.According to the pathological findings,21(45.7%)were proved to be polyp canceration,among which 6 were advanced carcinoma,and 3 were found metastasis to the lymph nodes.Among the 41 cases of laparoscopic resection,there was no conversion to an open surgery.Anastomotic leakage was found in 2 cases and anastomotic bleeding in 1.In the 5 cases of laparoscopic-assisted endoscopic polypectomy,no complication was observed.During the period of follow-up(1 to 21 months),no recurrence was detected.Conclusion Endoscopy combined with laparoscopy extends the safety and indications of endoscopic polypectomy,and is minimally invasive to the patients.It is an ideal procedure in the treatment of colorectal polyps and poly carceration.

This study aimed to assess the relationship of OAS2 rs739901 5'-flanking C/A polymorphisms with the susceptibility to Enterovirus-71 (EV71) infection.We investigated 294 hand-foot-mouth disease (HFMD) Chinese children with EV71 infection (165 mild cases and 129 encephalitis cases).The improved multiplex ligation detection reaction (iMLDR) technique was used to test the genotypes.In EV71-infected patients,the CA genotype distribution (P=0.007),A allele frequency (OR 1.32,95％ CI 1.0-1.7,P=0.034)and CA+AA carriage frequency (P=0.003) of OAS2 rs739901 5'-flanking were obviously elevated as compared with controls,but there were no statistically significant differences between mild cases and encephalitis cases.In EV71-infected patients,the counts of white blood cells (P=0.034) and blood glucose concentrations (P=0.042) were raised in A carriers (CA+AA).Among different genotypes of encephalitis cases,the contents of cerebrospinal fluid (CSF) showed no significant differences.IFN-γ levels in EV71-infected patients were higher than those in controls (mild group vs.control group,P＜0.01;encephalitis group vs.control group,P＜0.001).In encephalitis cases,IFN-γ levels were reduced (P＜0.05) in A carriers compared to CC genotype,however,there were no significant differences between genotypes CA and AA (P=0.226).These findings suggest that OAS2 rs739901 5'-flanking C/A genetic polymorphisms involve the susceptibility to EV71 infection,and A allele might be a risk factor of the susceptibility to EV-71 infection.