[Overexpression of miR-431-5p impairs mitochondrial function and induces apoptosis in gastric cancer cells via the Bax/Bcl-2/caspase3 pathway].

OBJECTIVE: To investigate the expression of microRNA miR-431-5p in gastric cancer (GC) tissues and its effects on apoptosis and mitochondrial function in GC cells. METHODS: The expression level of miR-431-5p in 50 clinical samples of GC tissues and paired adjacent tissues was detected using real-time fluorescence quantitative PCR, and its correlation with the clinicopathological features of the patients was analyzed. A cultured human GC cell line (MKN-45 cells) were transfected with a miR-431-5p mimic or a negative control sequence, and the cell proliferation, apoptosis, mitochondrial number, mitochondrial potential, mitochondrial permeability transition pore (mPTP), reactive oxygen species (ROS) production and adenosine triphosphate (ATP) content were detected using CCK-8 assay, flow cytometry, fluorescent probe label, or ATP detection kit. The changes in the expression levels of the apoptotic proteins in the cells were detected with Western blotting. RESULTS: The expression level of miR-431-5p was significantly lower in GC tissues than in the adjacent tissues (P < 0.001) and was significantly correlated with tumor differentiation (P=0.0227), T stage (P=0.0184), N stage (P=0.0005), TNM stage (P=0.0414) and vascular invasion (P=0.0107). In MKN-45 cells, overexpression of miR-431-5p obviously inhibited cell proliferation and induced cell apoptosis, causing also mitochondrial function impairment as shown by reduced mitochondrial number, lowered mitochondrial potential, increased mPTP opening, increased ROS production and reduced ATP content. Overexpression of miR-431-5p significantly downregulated the expression of Bcl-2 and increased the expressions of pro-apoptotic proteins p53, Bcl-2 and cleaved caspase-3 protein. CONCLUSION: The expression of miR-431-5p is down-regulated in GC, which results in mitochondrial function impairment and promotes cell apoptosis by activating the Bax/Bcl-2/caspase3 signaling pathway, suggesting the potential role of miR-431-5p in targeted therapy for GC.

[Xanthoceras sorbifolium Bunge flower extract inhibits benign prostatic hyperplasia in rats].

OBJECTIVE: To assess the inhibitory effect of the extract of Xanthoceras sorbifolium Bunge flower against benign prostatic hyperplasia (BPH) and explore its possible mechanism. METHODS: MTT assay was used to examine the effect of the extract of Xanthoceras sorbifolium Bunge flower on proliferation of benign prostatic hyperplasia cells (BPH-1), and cell apoptosis and cell cycle changes following the treatment were analyzed using annexin V/PI double staining and flow cytometry. The protein expression levels of Bcl-2, Bax, caspase-3, PI3K and AKT in the treated cells were detected using Western blotting. A rat model of BPH established by subcutaneous injection of testosterone propionate was treated with the flower extract for 28 days, and pathological changes in the prostate tissue were observed with HE staining. The protein expression levels of Bcl-2, Bax, caspase3 and PI3K/AKT in the prostate tissue were detected with Western blotting. RESULTS: Within the concentration range of 125-1000 µg/mL, the flower extract of Xanthoceras sorbifolium Bunge significantly inhibited the proliferation of BPH-1 cells and caused obvious cell cycle arrest at G0/G1 phase; the apoptotic rate of the cells was positively correlated with the concentration of the flower extract (P < 0.05). Bcl-2, p-PI3K and p-AKT expression levels were significantly down-regulated and Bax and caspase-3 expression levels were significantly increased in the cells after treatment with the flowers extract (P < 0.05). In the rat models of BPH, the rats treated with the flowers extract at moderate and high doses showed obviously decreased expressions of p-AKT and Bcl-2 and an increased expression of Bax in the prostate tissue; a significantly lowered p-AKT expression was observed in the prostate tissue of rats receiving the low-dose treatment (P < 0.05). CONCLUSION: The flower extract of Xanthoceras sorbifolium Bunge has a inhibitory effect on BPH both in vitro and in rats, suggesting its potential value in the development of medicinal plant preparations for treatment of BPH.

[A pretest model of obstructive coronary artery disease based on machine learning: from the C-Strat study].

Objective: To develop a pretest probability model of obstructive coronary artery disease with machine learning based on multi-site Chinese population data. Methods: Chinese regiStry in early deTection and Risk strAtificaTion of coronary plaques (C-Strat) study is a prospective multi-center cohort study, in which consecutive patients with suspected obstructive coronary artery disease and ≥64 detector row coronary computed tomography angioplasty (CCTA) evaluation were included. Data from the patients were randomly split into a training set (70%) and a test set (30%). More than 50% of coronary artery stenosis by CCTA was defined as positive outcome. A boosted ensemble algorithm (XGBoost), 10-fold cross-validation and Bayesian optimization were used to establish a new prediction model-CARDIACS(pretest probability model from Chinese registry in eARly Detection and rIsk stratificAtion of Coronary plaques Study), and a logistic regression was used to establish a model-LOGISTIC in training set. The test set was used for validation and comparison among CARDIACS, LOGISTIC, UDFM (updated Diamond-Forrester Model) and DFCASS(Diamond-Forrester and CASS). Results: The study population included 29 455 patients with age of (57.0±9.7) years and 44.8% women, of whom 19.1% (5 622/29 455) had obstructive coronary artery disease. For CARDIACS, the age, the reason for visit and the body mass index (BMI) were the most important predictive variables. In the independent test set, the area under the curve (AUC) of CARDIACS was 0.72 (95%CI 0.70-0.73), which was significantly superior to that of LOGISTIC (AUC 0.69, 95%CI 0.68-0.71, P=0.015), UDFM (AUC 0.64, 95%CI 0.62-0.65, P<0.001) and DFCASS (AUC 0.66, 95%CI 0.64-0.67, P<0.001), respectively. Conclusion: Based on Chinese population, the study developed a new pretest probability model--CARDIACS, which was superior to the traditional models. CARDIACS is expected to assist in the clinical decision-making for patients with stable chest pain.

[Correlation between CTL and Th1 cell proportion in peripheral blood of liver transplantation recipients and the success of hepatitis B vaccination].

Objective: To investigate the correlation between the proportion of CTL and Th1 cells in peripheral blood of liver transplant recipients and the success of hepatitis B vaccination. Methods: The subjects of this study were liver transplantation recipients with chronic HBV-related liver diseases in Organ transplantation institute of the third medical center of PLA general hospital. Subjects were randomly divided into two groups for prospective study. In the rapid group, one dose of 40 µg hepatitis B vaccine was administered at 0, 1, 2and 3 months, and one dose of 20 µg hepatitis B vaccine was administered at 4, 5 and 6 months. In the rapid-enhanced group, one dose of 40 µg hepatitis B vaccine was administered at 0, 1, 2 and 3 months, and one dose of 60 µg hepatitis B vaccine was administered at 4, 5 and 6months. Compare and analyze the success rate of inoculation, the titer of hepatitis B surface antibody (anti-HBs), the proportion of CTL cells in CD8(+)T cells and Th1 cells in CD4(+)T cells. Correlation analysis was performed for CTL and Th1 cells and anti-HBs, Observe the safety of vaccination. Results: The inoculation success rate, anti-HBs growth rate, CTL cell percentage increase and Th1 cell percentage increase in the rapid enhancement group were all higher than those in the rapid enhancement group, and the differences were statistically significant, they were 38.3% (23/60) vs 21.7% (13/60) (P=0.046), 91.3(72.5,124.2) vs 22.1(12.4, 31.6) (P=0.001), 1.4(0.8,1.9) vs 0.4(0.2,1.4) (P=0.001) and 7.4±2.6 vs 5.6±3.7 (P=0.001) respectively. The percentage increase of CTL cells and Th1 cells in the successful group was greater than that in the non-successful group, and the difference was statistically significant. They were 1.9(1.4,2.5) vs 0.1(0.0,1.1) (P=0.024) and 9.6±3.1 vs 2.4±2.0 (P<0.001). There was no significant correlation between anti-HBs increase (105.5±37.1) and CTL increase 1(0,3) (P=0.099), while there was significant positive correlation with Th1 increase 7(2,11) (P<0.001). No rejection reaction occurred during the study period, and there was no special abnormal change in the safety index. Conclusion: Reasonable increase of vaccine dose can up-regulate Th1 cell expression and promote the generation of anti-HBs.

[Twoyear outcomes and the influence factors of indirect pulp treatment in primary teeth: a retrospective study].

OBJECTIVE: To evaluate the outcomes and to find out the influence factors of indirect pulp treatment in primary teeth. METHODS: Children who received indirect pulp treatment in primary teeth in the Department of Pediatric Dentistry in Peking University School and Hospital of Stomatology from August, 2014 to September, 2016 were collected in the Electronic Medical Record Database of Peking University School and Hospital of Stomatology, via the database for scientific research provided by the corporation of Kaientai. The children selected as the subjects of this study were followed up over 1.5 years, and they were under 9 years old if in the group of primary molars while the children in the group of primary anterior teeth were under 4 and a half years old. Those children who were not reviewed regularly or didn't have complete medical records were removed. Basic information, the relevant medical records and radiographic records of those children were collected. All teeth were examined clinically and classified into 2 outcomes, teeth in group H were regarded as succeeded, and teeth in group P were regarded as failed. Survival analysis was applied. The survival rate and survival time of the deciduous teeth calculated. Multivariate analysis was performed by using Cox proportional hazard model. RESULTS: One hundred and six children were finally included, aged from 1.6 to 8.8 years, with the mean age of (5.0±1.7) years. 168 primary teeth (122 primary molars, 46 primary anterior teeth) were included, and the average follow up time was (729±244) days. Thirty-five primary teeth (23 primary molars, 12 primary anterior teeth) failed upon clinical or radiographic examinations by September, 2018. The cumulative survival probability of half a year, one year, one year and a half, two years, two and a half years for the indirect pulp treatment was 93.5%, 92.9%, 87.5%, 82.7%, and 75.5% through the KaplanMeier method, respectively. Through the analysis of Cox proportional hazard model, in primary molars, the survival probability tended to be lower when the number of tooth surface affected by caries was greater (OR=1.709, P<0.05). Compared with primary molars, the survival probability of primary anterior teeth was lower, but the difference was not significant. CONCLUSION: Complying the current instructions in our department, the cumulative survival probability of two and a half years after the indirect pulp treatment in primary teeth was 75.5%. In primary molars, the survival probability tended to be lower when the number of tooth surfaces affected by caries increased.

[Identification of yeasts from commercial attiéké in Abidjan (Côte d'Ivoire): Preliminary study]. / Identification des espèces de levures isolées de l'attiéké commercialisé sur les marchés à Abidjan (Côte d'Ivoire) : étude préliminaire.

OBJECTIVE: This study aims to identify yeasts from commercial attiéké. PATIENTS AND METHODS: This descriptive cross-sectional survey involving 103 samples of attiéké sold on the market from August to September 2017. It was conducted from the mycology laboratory of the Institut Pasteur of Côte d'Ivoire. The samples were seeded on YGC medium. The colonies of positive culture were counted and seeded again on chromogen medium. Identification of yeasts species was carried out by MALDI-TOF mass spectrometry (Vitek MS bioMérieux). RESULTS: As a whole, 63 samples were contaminated with fungi i.e. an overall prevalence of 61.1 %. Fifty-nine of yeasts were identified i.e. 57.2 %. Candida rugosa was most frequently isolated (62.7 %), followed by Ccandida albicans (8.5 %), Candida krusei (8.5 %) et Candida tropicalis (8.5 %). Geotrichum candidum represented 6.8 % isolated yeasts. The yeast average load is 10,124×102 CFU/g with high contamination of commercial attiéké in the commune of Abobo. CONCLUSION: The contamination of attiéké by yeasts is a reality. C. rugosa was most frequently isolated. It would be necessary to investigate the food chain to determine the source of contamination.

[Two-year outcomes of pulpotomy in primary molars using mineral trioxide aggregate: a retrospective study].

OBJECTIVE: To evaluate the two-year outcomes of primary molars pulpotomy using mineral trioxided aggregate (MTA) and to find out the potential influence factor, with the help of electronic medical record database. METHODS: Children who received primary molars pulpotomy in the Department of Pediatric Dentistry in Peking University School and Hospital of Stomatology from May, 2014 to November, 2015 were searched in the Electronic Medical Record Database of Peking University School and Hospital of Stomatology, via the database for scientific research provided by the corporation of Kaientai. The children who were healthy, no more than 8 years old and followed up over 1.5 years were selected as the subjects of this study. At the same time, those children who didn't have complete medical records and X-rays before and after treatment were removed. Basic information, the relevant medical records and radiographic records of those children were collected. All molars were examined clinically and radiographically, and classified into 1 of 5 outcomes: N, H, P0, PX, PY. Molars classified into P0, PX and PY were regarded as failed. Survival analysis was applied. The survival rate and survival time of the deciduous teeth were calculated.Multivariate analysis was performed by using Cox proportional hazard model. RESULTS: One hundred and fifteen children were finally included,aged from 2.6 to 8.2 years, with the mean age of (4.5±1.1) years. 211 primary molars were included, and the average follow up time was (880±154) days. A good level of agreement between the raters was found for molars with five outcomes(κ=0.913). Intrarater reliability was good for molars with five outcomes (κ=0.916). Forty-nine molars failed by September, 2017. Forty-three molars had abnormal radiographic manifestation. Six molars suffered premature loss. Only fourteen molars had an associated gingival swelling or parulis, or pathologic mobility upon clinical examination. The cumulative survival probability of half a year, one year, one year and a half, two years, two year and a half for the pulpotomy was 100%, 98.5%, 92.9%, 90.5%,73.8% through the Kaplan-Meier method, respectively. Through the analysis of Cox proportional hazard model, the survival probability was significantly higher when the age was younger. When sodium hypochlorite and the saline were used as the irrigation at the same time, the survival probability was not improved for those teeth with more bleeding in the operation compared with that used saline only. The teeth with preformed metal crown (PMC) gained longer median survival time than those restored with resin composite and others, but the difference was not significant, either. What's more, gender, the restoration time of PMC, the restoration of the opposite teeth were not found to be associated with the success of the treatment. CONCLUSION: The cumulative survival probability of two years after the primary pulpotomy was 90.5%, complying the current instructions in our department. The survival probability of the molars after the treatment was significantly affected by the age.

[A pilot study on the noninvasive fluid hemodynamic investigation of coronary plaque].

Objective: To characterize the hemodynamic force towards coronary plaque based on noninvasive coronary computed tomographic angiography and to investigate its relationship with plaque features and stenosis severity by computational fluid dynamics. Methods: Twenty-six patients underwent invasive fractional flow reserve measurement following coronary computed tomography angiography examination from March to September 2016 were retrospectively included. Computational fluid dynamics was applied and wall shear stress (WSS) and axial plaque stress (APS), which extracted the axial component of hemodynamic stress acting on stenotic lesions, were calculated based on the results of noninvasive coronary computed tomographic angiography. Plaque analysis was performed to elucidate plaque features and relative plaque burden. The fluid dynamics distributions in lesions with different stenosis severity were investigated. Results: Thirty-one coronary plaques with satisfactory imaging quality were analyzed, there were 11 (35.5%) dominant low WSS (<1 Pa) lesion and 20 high WSS lesion (64.5%), 8(25.8%) net retrograde APS lesion and 23(74.2%) anterograde lesion. Plaque volume was (78.5±48.6) mm(3) and plaque burden was (69.1±12.1)% in the low WSS group, which was(60.5±57.3) mm(3), and(57.5±14.0)%, respectively in the high WSS group, the plaque burden was significantly higher in the low WSS group than in the high WSS group (P=0.028), while the percentage of calcified plaque, fibrotic plaque and lipid core volume were similar between the two groups (P>0.05). Plaque volume was (79.7±69.1) mm(3) and plaque burden was(68.7±13.7)% in the group with anterograde-dominant APS plaque, which was(61.7±24.9)mm(3), and(68.9±10.4)%, respectively in the net retrograde APS lesion group (P>0.05). Percentage of lipid core area was significantly higher in the anterograde lesion group than in the retrograde lesion group ((25.1±18.1)% vs.(10.8±12.7)%, P=0.049). Both WSS and APS were significant higher in the severe obstructive coronary stenosis group than in non-severe obstructive coronary stenosis group (P<0.05). Although there was no difference in WSS between functional coronary ischemia group and non-functional coronary ischemia group ( (13.3±8.7) Pa vs. (12.5±14.2) Pa, P>0.05), the distribution of APS was different between the functional coronary ischemia group and non-functional coronary ischemia group ((1 698.8±652.6) Pa vs. (981.4±787.5) Pa, P<0.05). Conclusion: WSS and APS can uniquely characterize the stenotic segment and has a strong relationship with lesion geometry. APS may be related to the necrotic core plaque and functional coronary ischemia. Clinical application of these hemodynamic and geometric indices may be helpful to assess the future risk of plaque progress and plaque rupture, which will be helpful on determining respective treatment strategy for patients with coronary artery disease.

In Vitro and In Vivo Characterizations of Chiglitazar, a Newly Identified PPAR Pan-Agonist.

Solid rationales are still present for the identification of synthetic ligands to simultaneously target multiple PPAR subtypes for the treatment of T2DM. The purpose of this study was to characterize the in vitro and in vivo differential effects of chiglitazar, a non-TZD type of PPAR pan-agonist currently in phase III clinic development in China, from PPARγ-selective agonist like rosiglitazone. Chiglitazar showed transactivating activity in each PPARα, γ, and δ subtype and upregulated the expression of PPARα and/or PPARδ downstream genes involved in the key processes of lipid metabolism and thermogenesis. Comparable blood glucose lowering effect was observed between chiglitazar and rosiglitazone, but chiglitazar did not significantly increase the body weight in KKAy and fat pad weight in db/db mice. Chiglitazar had high distribution in liver, pancreas, and skeleton muscles but was less present in kidney, heart, and adipose in rats. Heart weight increase was not observed in rats treated with chiglitazar for 6 months at a dose as high as 45 mg kg(-1). The in vitro and in vivo differential features of chiglitazar are informative and encouraging for the further development of this synthetic ligand for the potential use in T2DM.

Pharmacokinetics and metabolism of 1,5-dicaffeoylquinic acid in rats following a single intravenous administration.

1,5-Dicaffeoylquinic acid (1,5-DCQA) is a potentially important HIV-1 integrase inhibitor widely distributed in many plants. To characterize the pharmacokinetic and metabolic properties of 1,5-DCQA in rats following single intravenous administration (160 mg/kg), the plasma concentrations of 1,5-DCQA were measured by high-performance liquid chromatography (HPLC) and the metabolites formed in urine were identified by liquid chromatography-mass spectrometry (LC-MS) in parallel to diode-array detection (DAD). The results showed that the concentrations of 1,5-DCQA in plasma declined rapidly in a biphasic manner with a mean terminal half-life (t(1/2)) of 1.40 h. The mean clearance (CL) and the apparent volume of distribution (Vd(B)) of 1,5-DCQA were 0.44l/h/kg and 0.89l/kg, respectively. A total of 15 metabolites in rat urine were identified, including four isomeric O-mono-methylated (M1-M4), six isomeric O-di-methylated (M5-M10), one isomeric O-mono-methyl-glucuronidated (M11) and four isomeric O-di-methyl-glucuronidated (M12-M15) metabolites. The O-methylation positions of three important metabolites (M1, M2 and M5) were determined (3''-, 3'-, and 3',3''-) by comparing with synthesized standards. These results suggested that the disappearance of 1,5-DCQA from plasma was rapid, and that its quick urinary excretion and extensive metabolism, including methylation and glucuronidation, were two factors causing its rapid elimination from the circulation.

[Investigation on primary vectors of Borrelia burgdorferi in China].

From 1987 to 1997, primary vectors of borrelia burgdorferi sensu lato (B.b.s.l.) were investigated in China. A total number of 17,000 ticks collected from 20 provinces, city and autonomous region were classified, including 2 families 8 genus 23 species. There were 108 isolates of B.b.s.l. discovered from 8 species of ticks, including Ixodes persulcatus, I. granulatus, I. acutitarsus, Haemaphysalis concinna, H. longicornis, H. bispinosis, H. cornigera taiwana and Dermacentor silvarum. Ixodes persulcatus was found as the dominant species, accounted for more than 80% of the total number of collected ticks. 20 to 45 per cent adult Ixodes persulcatus contained spirochetes as determined by direct immunofluorescence in the northeast and northwest areas. Seasonal change of adult Ixodes persulcatus was coincident with patients with erythema chronicum migrans. These studies proved that Ixodes persulcatus played a leading role in the transmission of B.b.s.l. to human being in northern region of China. Haemaphysalis bispinosis and Ixodes granulatus were identified as the dominant species with 16 to 40 per cent adult Haemaphysalis bispinosis and 24% per cent adult Ixodes granulatus containing spirochetes as determined by direct immunofluorescence in the southern region of China. Haemaphysalis bispinosis and Ixodes granulatus were proved to be important vectors of B.b.s.l. in the southern area of China.