Objective To investigate the molecular mechanism underlying Porphyromonas gingivalis(Pg)-induced autophagy in esophageal squamous cell carcinoma(ESCC).Methods After Pg infected KYSE70 cells and KYSE140 cells pretreated with siAtg7 or Chloroquin(CQ),Western blot was used to measure protein levels of Atg7,LC3-Ⅱ/LC3-Ⅰ,and p62;Immunofluorescent confocal imaging analysis was used to detect autophagosome and autolysosome;CCK-8 assay was used to test cell viability;Transwell assay was used for ESCC cell migration and invasion potentials.Likewise,miR-21-5p inhibitor,RASA1 overexpression plasmid,or U0126 were used to block miR-21-5p/RASA1/ERK signaling pathway prior to Pg infection,followed by the aforementioned methods.In addition,immunohistochemistry was used to examine Pg abundance and LC3 protein levels,and RT-PCR was used to evaluate miR-21-5p expression in ESCC and adjacent tissue samples,followed by correlation analyses be-tween Pg and LC3,and Pg and miR-21-5p.Results Pg infected KYSE70 cells and KYSE140 cells showed upreg-ulation Atg7 protein and LC3-Ⅱ/LC3-Ⅰ protein but downregulation of RASA1 protein and p62 protein,enhanced cell proliferation,migration,and invasion as well as immunofluorescent spots of red,green,and yellow in mRFP-GFP-LC3-labeled ESCC cells.Pretreatment with CQ or siAtg7 abolished the above alterations induced by Pg.Con-sistently,pretreatment with miR-21-5p inhibitor,U0126,or RASA1 overexpression plasmid also blocked Pg-stimu-lated autophagy.In ESCC samples,Pg abundance was correlated with upregulation of miR-21-5p and LC3.Con-clusion Pg promotes autophagy in esophageal squamous cell carcinoma via miR-21-5p/RASA1/ERK signaling pathway.

OBJECTIVETo detect potential mutations in two neonates suspected for Cornelia de Lange syndrome (CdLS).

METHODSPeripheral blood samples from the neonates and their parents were collected and analyzed for CdLS-related genes using targeted sequence capture and next-generation sequencing. Suspected mutations were confirmed by direct Sanger sequencing.

RESULTSThe neonates were found to respectively carry mutations c.7219C to T and p.D2339Lfs*4 of the NIPBL gene, among which the p.D2339Lfs*4 mutation has not been reported previously. No pathogenic mutation was found in other CdLS-related genes including NIPBL, SMC1A, SMC3, RAD21 and HDAC8.

CONCLUSIONThe c.7219C to T and p.D2339Lfs*4 mutations of the NIPBL gene probably account for the disease in both patients.

Objective:To analyze the relationship between plasma fibrinogen(FIB) within normal range and microalbuminuria in elderly patients with type 2 diabetes mellitus.Methods:A total of 869 elderly subjects with type 2 diabetes mellitus admitted to the Department of Endocrinology of Shanghai Fifth People′s Hospital from October 2012 to October 2014 were included in the study. The patients were divided into four groups based on the quartile level of FIB: Q1 group(<2.42 g/L), Q2 group(2.42-2.89 g/L), Q3 group(2.90-3.61 g/L), and Q4 group(≥3.62 g/L). The relationship between FIB and urinary albumin/creatinine ratio(UACR) was analyzed.Results:With the increasing of FIB, the level of UACR was significantly elevated( P<0.05). Pearson correlation analysis showed that FIB was positively associated with age, duration of diabetes, creatinine(Cr) and UACR in men and women( P<0.01). Multiple regression analysis showed that FIB was an independent factor of UACR( P<0.01). Logistic regression analysis showed that the risks of microalbuminuria and macroalbuminuria were respectively 4.536 folds(95% CI 2.516-8.175, P<0.01) and 13.314 folds(95% CI 2.925-60.612, P<0.01) in Q4 group, and 2.177 folds(95% CI 1.273-3.724, P<0.01) and 4.098 folds(95% CI 1.101-19.226, P<0.05) in Q3 group as compared with Q1 group after adjused by following factors: gender, age, duration of diabetes, body mass index(BMI), systolic blood pressure(SBP), diastolic blood pressure(DBP), fasting plasma glucose(FPG), HbA 1C, total cholesterol(TC), triglyceride(TG), low density lipoprotein-cholesterol(LDL-C), Cr, alanine aminotransferase(ALT), as well as smoking and drinking behavior. Based on the cut off values to UACR 30 mg/g and 300 mg/g, the receiver operating characteristic curve(ROC) was used to evaluate the value of FIB for UACR. The optimal cut-off value of FIB was 3.18 g/L and 3.22 g/L respectively. Conclusions:Plasma FIB was closely associated with microalbuminuria in elderly patients with type 2 diabetes mellitus, which may be considered as one of the predictors for diabetic nephropathy.

Traditional Chinese Medicine (TCM) has certain advantages in the treatment of precancerous lesions of gastric cancer (PLGC) based on the holistic concept and the thought of syndrome differentiation. Currently, it is generally divided into 6 kinds of syndromes: liver and stomach qi stagnation syndrome, liver and stomach heat stagnation syndrome, spleen and stomach weakness syndrome (including spleen and stomach qi deficiency syndrome with coldness), spleen and stomach damp heat syndrome, stomach yin deficiency syndrome and blood stasis in stomach collateral syndrome. Clinically, the doctor should treat PLGC patients according to different syndrome types by using Chinese medicine prescription, which could improve the gastric mucosal pathological state, gastroscopy and clinical symptoms, to rehibit the development of precancerous lesions, reduce the incidence rate of gastric cancer. In the future, the doctors shouldreach the consensus of treating PLGC with TCM diagnosis, and focus on the research of TCM compounds or monomers with obvious curative effect, increase the times of follow-up, and evaluate the long-term curative effect.

Objective To investigate the mechanism underlying the suppressive effect of Porphyromonas gingivalis(P.gingivalis)on ferroptosis in esophageal squamous cell carcinoma(ESCC).Methods ESCC cells infected with P.gingivalis and uninfected control cells were treated with ferroptosis inducer RSL3 followed by measurements of cell viability,malondialde-hyde(MDA),reactive oxygen species(ROS),and the expression of glutathione peroxidase 4(GPX4).Moreover,the expression of hypoxia-inducible factor-1α(HIF-1α)and its target genes were detected by qPT-PCR,Western blotting or immunohistochem-istry in ESCC tissue and cells under the condition of P.gingivalis infection.The effect of P.gingivalis infection combined with the HIF-1α inhibitors LW6 and RSL3 on ferroptosis in ESCC was detected in vitro and in vivo.Results P.gingivalis infection of the ESCC cells resulted in an increase of the cell viability(P＜0.05),decreased levels of intracellular ROS(P＜0.05)and MDA(P＜0.05)and increased the expression of GPX4 compared with RSL3 treatment alone.In ESCC tissues,the increased a-bundance of P.gingivalis was correlated with upregulation of HIF-1α.Furthermore,P.gingivalis infection induced upregula-tion of HIF-1α and its target genes.LW6 promoted ferroptosis via inhibiting the HIF-1α upregulation induced by P.gingivalis infection in vitro and in vivo.Conclusion HIF-1α renders resistance to ferroptosis in P.gingivalis infected ESCC.Combination of HIF-1α inhibitory agents and ferroptosis inducing agents might be a novel therapeutic strategy in ESCC care.

ObjectiveTo observe the clinical effectiveness and safety of Dingchan Granule （定颤颗粒） for paroxysmal atrial fibrillation with syndrome of qi stagnation and blood stasis. MethodsUsing a randomised， double-blind， placebo controlled study method， 90 patients with paroxysmal atrial fibrillation with qi stagnation and blood stasis syndrome were divided into 45 cases each in the treatment group and the control group. Both groups were given conventional western medicine treatment， and the treatment group was additionally treated with Dingchan Granule， while the control group was treated with Dingchan Granule placebo， both of which were taken orally for 8 g each time twice a day. Both groups were treated for 8 weeks. We compared the clinical effectiveness， the improvement of traditional Chinese medicine （TCM） symptoms and the recovery rate of atrial fibrillation between the two groups. We compared the number and duration of atrial fibrillation episodes， TCM symptoms score， atrial fibrillation symptom classification， 24-hour average ventricular rate， Pittsburgh Sleep Quality Index （PSQI）， anxiety index， depression index before and after treatment， and evaluated the safety of the two groups. ResultsThe total clinical effectiveness rate in the treatment group was 82.22% （37/45）， which was better than 60.00% （27/45） in the control group （P＜0.05）. The total effective rate of TCM syndrome effectiveness in the treatment group was 88.89% （40/45）， which was better than 66.67% （30/45） in the control group （P＜0.05）； and the rate of atrial fibrillation regression in the treatment group was 26.67% （12/45）， better than 6.67% （3/45） in the control group （P＜0.05）. The number and duration of atrial fibrillation episodes in both groups were significantly decreased （P＜0.01）， and the number and duration of atrial fibrillation episodes in the treatment group were lower than those in the control group （P＜0.01）. The TCM syndrome scores of both groups after treatment were significantly lower than before treatment （P＜0.01）， and the scores of the treatment group was lower than those of the control group （P＜0.05）. The severity of atrial fibrillation symptoms and the grading of atrial fibrillation symptoms in both groups after treatment were improved （P＜0.01）， and the degree of symptom improvement in the treatment group was better than that in the control group （P＜0.01）. The 24-hour average ventricular rate of both groups after treatment was significantly lower （P＜0.01）. The PSQI， anxiety index and depression index of the treatment group were all lower than before treatment （P＜0.01）， while the PSQI and anxiety index of the control group were both lower than before treatment （P＜0.01 or P＜0.05）， the PSQI， anxiety index and depression index of the treatment group being lower than those of the control group （P＜0.05 or P＜0.01）. No adverse events occurred in both groups， and no abnormalities were observed in blood， urine， stool routine， liver and kidney function， and coagulation function indexes. ConclusionDingchan Granule for paroxysmal atrial fibrillation with qi stagnation and blood stasis syndrome can alleviate clinical symptom， improve TCM symptom scores， increase atrial fibrillation recovery rate， stabilise the average ventricular rate， and significantly improve the quality of sleep， alleviate the anxiety and depression， with a good safety profile.