Serum metabolomics analysis and establishment of diagnostic model of pancreatic cancer associated diabetes / 中华消化杂志

Objective To establish the diagnostic model based on detection of serum biomarkers in pancreatic cancer (PC) associated diabetes.Methods From June 2013 to July 2014, at Ruijin Hospital, School of Medicine , Shanghai Jiao Tong University , 30 patients diagnosed with PC companied with new onset diabetic mellitus and 30 patients with new onset type 2 diabetic mellitus , were enrolled .Serum samples were examined by liquid chromatography-mass spectrometry ( LC-MS) for metabolomics analysis .Orthogonal partial least square ( OPLS ) was performed for raw data analysis to obtain the differentially expressed metabolites between two groups .The first 15 cases of each group were taken as training samples and the left as validation samples.The model was established using logistic regression via stepwise differentially expressed metabolites and clinical data input in training samples .The diagnostic efficiency of the model was verified in validating samples . Results Ten differentially expressed metabolites were identified in PC companied with new onset diabetic mellitus group and new onset type 2 diabetic mellitus group .The differentially expressed metabolites identified in positive ion mode were 3-ketosphingosine , arachidonoyl dopamine , phosphatidylethanolamine ( 18 :2 ) , ubiquinone-1 and valine .The differentially expressed metabolites identified in negative ion mode were C 16 sphingosine-1-phosphate, keto palmitic acid, isoleucine, N-succinyl-L-diaminopimelic acid and uridine.The diagnostic model was established in training samples:p=e(Xβ)/(1+e(Xβ)), ( Xβ) =-158.975-1.891 (age) +0.309 ( phosphatidylethanolamine 18:2 ) +1.035 ( C16 sphingosine-1-phosphate ) +0.084 (isoleucine) +1.1145 ( N-succinyl-L-diaminopimelic acid ).The area under curve ( AUC) of receiver operating characteristic (ROC) of this model was 0.982 in validation samples, the sensitivity and specificity were both 93.3％.Conclusion Serum metabolomics-based diagnostic approach is a promising method for screening PC from new onset diabetic mellitus .

Construction of human mucosa oral epithelial cell lines overexpressing telomerase reverse transcriptase gene mediated by lentivirus / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To construct a cell line of oral mucosa epithelial cells that stably express human telomerase reverse transcriptase (hTERT) by lentiviral vectors, approaches for the establishment of stable and efficient immortalized oral mucosa epithelial cell lines were explored.</p><p><b>METHODS</b>Whole RNA was extracted from 293T cells. The hTERT gene was amplified by polymerase chain reaction (PCR) and cloned into the lentiviral vector as pLVX-puro-hTERT. The lentivirus particles were successfully packaged and used to infect primary oral epithelial cells. The positive cell clones were selected by puromycin. Finally, the expression of hTERT was examined by real-time fluorescent quantitative PCR (qRT-PCR) and Western blot analysis.</p><p><b>RESULTS</b>The sequencing results confirmed the construction of the recombinant lentivirus pLVX-puro-hTERT. The morphology of infected cells was similar to that of normal oral mucosal epithelial cells, with a cobble stone-like appearance. The qRT-PCR and Western blot results showed that hTERT was overexpressed in infected cells compared with the normal group (P<0.05).</p><p><b>CONCLUSIONS</b>The oral epithelial cell line with stable expression of hTERT was successfully established by the lentivirus, which provides an experimental basis for the establishment of a highly efficient and stable oral epithelial immortalized cell line.</p>

Biological characteristics of a human specifically targeted antimicrobial peptide C16LL-37 against Streptococcus mutans / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This study aimed to evaluate the biological characteristics of a human specifically targeted antimi- crobial peptide C16LL-37 against Streptococcus mutans (S. mutans).</p><p><b>METHODS</b>In this study, an antimicrobial peptide LL-37, a peptide derived from CSP(C16) (S. mutans competence stimulating peptide), and recombinant peptide C16LL-37 were synthesized by Fmoc-chemistry-based strategy. The selectivity and antibacterial activity of C16LL-37 were identified by the colony counting method on microbial culture plates. After treatment of C16LL-37 at 32 µmol · L⁻¹, the morphological changes in S. mutans were observed by using scanning electron microscopy (SEM). In addition, enzyme-linked immunosorbent assay was used to evaluate the hemolytic activity and antibacterial activity of C16LL-37 under different conditions.</p><p><b>RESULTS</b>1) The minimum inhibitory concentration of C16LL-37 was 16 µmol · L⁻¹, and the minimum bactericidal concentration was 64 μmol ·L⁻¹. 2) The survival rate of S. mutans was 3.46% after C16LL-37 treatment at 64 µmo-L⁻¹ for 30 min, whereas it was 0% at 64 µmol · L⁻¹ for 60 min. The survival rates of four other kinds of bacteria were more than 60% at any time (P < 0.05). 3) The morphological change in S. mutans was observed after C16LL-37 treatment at 32 µmol · L⁻¹ by using SEM. S. mutans presented an irregular shape, rough surface, and evident splitting. 4) The hemolysis rate of C16LL-37 (≤ 64 µmol · L⁻¹) was less than 0.33%. 5) This study showed no significant in- fluence on the antibacterial activity of C16LL-37 under different conditions, such as temperature, pH, salinity, and trypsin at low concentration (P > 0.05).</p><p><b>CONCLUSION</b>C16LL-37 exhibited obvious specificity for S. mutans, strong antibacterial activity, low toxicity, and high stability. Thus, C16LL-37 has good potential in caries research and clinical application.</p>

Research advances in relationship between diabetes and pancreatic cancer / 临床肝胆病杂志

Pancreatic cancer (PC)is one of common malignant digestive diseases.It is mostly diagnosed at advanced stage,with an ex-tremely poor progression.The relationship between diabetes and PC was shown by numerous epidemiological studies for decades.Retrospec-tive clinical studies and research on molecular mechanisms in recent years have led to a new understanding of the relationship between diabe-tes,especially new-onset diabetes,and PC,the effect of antidiabetic medication on PC,and the molecular mechanisms underlying the con-nection between diabetes and PC.It is suggested by recent data that long-standing diabetes is one of the risk factors for PC development, new-onset diabetes may facilitate early diagnosis of PC,diabetes may have an impact on the prognosis of PC,the option of antidiabetic medication may influence the incidence of PC,and exploring the molecular mechanisms underlying the association between diabetes and PC may help to identify the new therapeutic target for PC.

Effect of EDA-A1 gene mutant on proliferation and cell cycle distribution of cultured human umbilical vein endothelial cell ECV304 / 重庆医学

Objective To investigate the effect of ectodysplasin A (EDA‐A1) gene of hypohidrotic ectodermal dysplasia on pro‐liferation and cell cycle of human umbilical vein endothelial cell (ECV304). Methods Recombinant eukaryotic expression vectors pcDNA3. 1(‐)‐EDA‐A1‐M /W (mutant, M and wild, W) containing the coding sequence were transected into ECV304 cells. EDA‐A1 gene was amplified by reverse transcription polymerase chain reaction (RT‐PCR), and the protein was detected by Western blot. Cell viability and cycle distribution were invested by MTT and Flow cytometry (FCM ). Results The EDA‐A1 gene and pro‐tein were detected respectively by RT‐PCR and Western blot in ECV cells transfected with pcDNA3. 1(‐)‐EDA‐A1‐M /W, but not in ECV cells transfected with plasmid pcDNA3. 1(‐) and cells without transection. And also, compared with control groups, EDA‐A1 gene mutant significantly decreased proliferation of ECV cells and its inhibition rate was 45. 70% ( P 0. 05). A significant increase of the G0 /G1 and S fraction was seen in the ECV cells of mutant group, compared with wild group with an accumulation in S phase and a concomitant decrease in G2 /M phase population (P< 0. 05). Conclusion Mutant and wild EDA‐A1 gene may have distinct biological functions on proliferation and cell cycle distribution of cultured human umbilical vein endothelial cell.

Analysis of the infection status of saliva Helicobacter pylori in Lanzhou / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To determine the prevalence of saliva Helicobacter pylori in Lanzhou and investigate Helicobacter pylori-related diseases.</p><p><b>METHODS</b>Helicobacter pylori was detected through bacterial culture, Gram stain microscopy, and urease test from saliva samples collected from 941 residents of Lanzhou. The infection rate and growth of Helicobacter pylori among the residents were analyzed in terms of different oral health conditions, oral disease, gender, urban and rural status, and age.</p><p><b>RESULTS</b>The rate of Helicobacter pylori-positive saliva in Lanzhou was 42.72%. The status of Helicobacter pylori infection showed significant difference among subjects with different oral hygiene and oral diseases. The rate of Helicobacter pylori-positive saliva among females was 47.89%, which was greater compared with the rate among males (38.45%, P = 0.004, chi2 = 8.492). The rate of Helicobacter pylori-positive saliva in the town was 33.99%, which was less than the rate for the villages (50.93%, P = 0.000, chi2 = 27.551). The rate of Helicobacter pylori-positive saliva among residents aged 10 to 59 showed a flat trend with no significant differences. However, the rate of Helicobacter pylori-positive saliva among residents over 60 years old showed a significant increase. No significant difference was found in the growth of saliva Helicobacter pylori (P = 0.086).</p><p><b>CONCLUSION</b>The rate of Helicobacter pylori-positive saliva is related to the subjects' oral hygiene, oral disease, gender, age, and living conditions.</p>

Effects of non-saccharomyces albicans metabolic products on the proliferation of human umbilical vein endothelial cell ECV304 / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the effects of non-Saccharomyces albicans metabolic products on the cell cycle distribution and proliferation of human umbilical vein endothelial cell ECV304 cells in vitro.</p><p><b>METHODS</b>The parallel dilution supernatant of Saccharomyces tropicalis, Saccharomyces krusei and Saccharomyces glabrata were prepared, and 1, 4, 16-fold(s) diluted concentration and control group were set up. The line of human umbilical vein endothelial cell ECV304 was cultured in vitro and treated by non-Saccharomyces albicans supernatant. The proliferous effect of ECV304 induced by non-Saccharomyces albicans supernatant after 24, 48, 72 h was detected by the methods of MTT, and the changes of cell density and cycle after 48 h were investigated by inverted microscope and flow cytometry.</p><p><b>RESULTS</b>At the 24th hour, all of the higher concentration (1-fold) of non-Saccharomyces albicans supernatant and the 4-folds diluted Saccharomyces krusei could promote ECV304 proliferation(P < 0.05). After adding various non-Saccharomyces albicans supernatant at 48h and 72th hour, Saccharomyces krusei supernatant and Saccharomyces glabrata supernatant significantly increased proliferation rate of ECV304, while Saccharomyces tropicalis supernatant group showed no significant change no matter which concentration was tested. At 48th hour after adding the non-Saccharomyces albicans supernatant, the ECV304 cells density treated by Saccharomyces krusei supernatant and Saccharomyces glabrata supernatant were significantly higher under the inverted microscope. The G0/G1 population of ECV304 cells decreased while cell proliferation index (PI) increased after incubated with Saccharomyces krusei supernatant and Saccharomyces glabrata supernatant for 48 hours (P < 0.05). Saccharomyces tropicalis group showed no significant change (P > 0.05).</p><p><b>CONCLUSION</b>The metabolic products of Sacharoymces krusei and Saccharomyces glabrata could induce proliferation of ECV304 cell, which suggests non-Saccharomyces albicans should be undergone more attention clinically in detection and treatment.</p>

A study on human tongue cancer cells' proliferation affected by Lactobacillus acidophilus / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the effects of Lactobacillus acidophilus (L. acidophilus) on the proliferation and cell cycle distribution of human tongue cancer cells (Tca8113 cells).</p><p><b>METHODS</b>In vitro cultivated human Tca8113 cells were treated by L. acidophilus supernatant, inactivated bacilli, cell free extracts and normal culture medium respectively, which were 1, 4, 16-fold(s) dilutelly, to investigate the proliferous effects of Tca8113 cells using of inverted microscope, cell counting, sulforhodamine B (SRB) and flow cytometry. The free radicals and Ca2+ in Tca8113 cells were also studied by confocal laser scanning microscope (CLSM).</p><p><b>RESULTS</b>At the 48th hour after adding different L. acidophilus components, the Tca8113 cells changed in shape from the diamond-like, polygonal and slabs into the elongated form. In the condition of different times and different culture concentrations, the proliferation of Tca8113 cells was significantly inhibited by L. acidophilus components, which enhanced as the time prolonged and the concentrations of each L. acidophilus components increased according to the cell counting and the SRB experimental analysis. The cell proliferation index (CPI) was significantly reduced (P<0.01). The free radicals and Ca2+ in Tca8113 cells under the effect of each L. acidophilus components for 48 h indicated an obviously rising (P<0.01).</p><p><b>CONCLUSION</b>L. acidophilus restrains the proliferation of Tca8113 cells, which might be due to the increase in quantity of free radicals and Ca2+ in Tca8113 cells, and might be resulted from the release of metabolic products of L. acidophilus.</p>

The correlation between oral colonization of Helicobacter pylori and gastrointestinal disease / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the correlation between Helicobacter pylori (H. pylori) colonization in the oral cavity and gastrointestinal disease.</p><p><b>METHODS</b>173 patients with gastrointestinal disease were grouped according to age, gender, periodontal status and types of gastrointestinal disease. H. pylori were detected from saliva samples of all patients by in vitro cultur. The H. pylori-positive rates in different groups were statistically analysed.</p><p><b>RESULTS</b>The H. pylori-positive rate in all patients was 40.46% and the difference between male and female showed significant (P<0.05). The H. pylori-positive rate was 56.72% in the age range 45-64, which was significantly higher than two younger age groups (P<0.05). The H. pylori-positive rate in patients with atrophic gastritis was 77.78%, of which the difference was significantly higher than superficial gastritis group and gastric and duodenal ulcer group respectively (P<0.05). The H. pylori-positive rate in healthy periodontia group was 15.38%, while that in periodontitis group was 72.73% (P<0.05).</p><p><b>CONCLUSION</b>H. pylori is a conditional pathogen. The H. pylori-positive rate from saliva is closely related to the types of gastrointestinal disease in patients, and it is correlated with the periodontal diseases as well. These findings suggest that the oral cavity with periodontal diseases is an ecological niche of H. pylori which might be an important cause for occurrence and re-occurrence of gastrointestinal disease.</p>

In vitro antibiotic effect of the leaching solution of astragalus on cariogenic bacteria / 实用口腔医学杂志

Astragalus produced in Gansu were chosen as the raw material to leachate. Studied the antibiotic effects of the leaching solution on the cariogenic bacteria and compared with the imported bacteriostatic product MI. Streptococcus mutans and Lactobacilli were cultured in the medium for 24 h. The PH and A600 values were measured. Statistical analysis was conducted by using SPSS 13.0. The leaching solution of astragalus has the same inhibitory effects on the growth and acid production of streptococcus mutans and lactobacilli as MI.

An epidemiologic study on candidal transmission from mothers to their neonates in Lanzhou / 实用口腔医学杂志

Objective: To investigate the candidal infection status in puerperas in Lanzhou, and the candidal transmission from mothers to their newborn infants. Methods: Vaginal fluid and saliva samples from 104 puerperas, as well as 104 saliva samples from their newborn infants were collected. The Candida species were cultured, isolated and identified using CHROMagar media. Further identification was done using molecular biological method. Results; In 81 of 312 specimens (104 x2 from mothers and 104 from infants), Candida species were found. 39.42% (41 cases) was observed in the vaginal fluid and 33.65% (35 cases) was in saliva of puerperas respectively, and 21. 15% (22 cases) in both vagina and oral cavity. 4.81% (5 cases) was found in oral cavities of newborn infants. The distribution of Candida species were 53 Candida albicans, 33 Candida glabrata, 2 Candida krusei and 1 Candida tropical. In 2 pairs of mother-infant, the same genotype of Candida ablicans was identified using PCR method. Conclusion; The Candida detection rate of newborn infants and transmission rate from mothers to their neonates in Lanzhou are higher than that reported in other areas. The colonization of Candida in newborn infants is relevant to both horizontal and vertical transmission. It can decrease the possibility of Candidal infection in newborn infants by controlling the Candidal transmission in hospital and preventing the infection in pregnant women.

Clinical significance and possible mechanism of DJ-1 expression in tumors / 国际肿瘤学杂志

DJ-1, ubiquitously expressed in various human tissues, is a multi-functional protein that plays roles in tumors, Parkinson' s disease, breeding and apoptosis. Over- expression of DJ-1 which is identified as a novel oncogene has been reported in several cancer cells. The underlying mechanisms include the anti-oxi-dative stress reaction, down-regulating phosphatase and tensin homolog deleted on chromosome 10 (PTEN) pathway and up-regulating Androgen pathway. However, there are also some evidences show low-expression of DJ-1 in tumor cells.