Effects of individualized antiplatelet therapy based on CYP2C19 genotype and platelet function on the prognosis of patients after PCI.

OBJECTIVE: To evaluate the effect of individualized antiplatelet therapy based on CYP2C19 genotype and platelet function on the prognosis of patients after percutaneous coronary intervention (PCI) compared with conventional antiplatelet therapy. PATIENTS AND METHODS: Patients diagnosed with acute coronary syndromes (ACS) in Shandong Provincial Qianfoshan Hospital from December 2014 to December 2017 were included in this prospective study and randomly divided into conventional (CA) and individualized antiplatelet therapy group (IA) at 1:1 ratio. Patients in the CA group received clopidogrel 75 mg once a day (QD). Group IA was divided into extensive, intermediate, and poor metabolizers according to the results of the CYP2C19 gene test. Three genotypes were given clopidogrel 75 mg QD, 75 mg twice daily (BID) and ticagrelor 90 mg BID respectively. After taking these medicines for a period of time, platelet function was monitored by thromboelastography (TEG) and MAADP values were recorded. MAADP indicates the adenosine diphosphate (ADP) induced platelet function that not inhibited by medicine. High platelet reactivity (HPR) was defined as MAADP > 47mm, indicating a high risk of thrombus, and MAADP ≤ 31 mm indicates a high risk of hemorrhage. For extensive metabolizers (EMs) and intermediate metabolizers (IMs) patients with HPR, the antiplatelet therapy would be changed by the clinician according to the patient's conditions. Major adverse cardiovascular events (MACE) and hemorrhage events were monitored during 1-year follow-up. RESULTS: The patients with MAADP > 47 mm were 89 (28.6%) in the IA group. There were 50 EMs patients with MAADP > 47 mm (33.3%). Of which, there were 2 cases which changed the dosage of clopidogrel to 75 mg BID, 14 cases who changed clopidogrel to ticagrelor. There were 36 IMs patients with MAADP > 47 mm (30.8%). Of which, there were 19 cases who changed clopidogrel to ticagrelor. There was no significant difference in the value of MAADP between EMs and IMs patients. Within 1 year after PCI, the occurrence of MACE in the IA group was significantly lower than that in the CA group (p=0.010). CONCLUSIONS: (1) Patients with a CYP2C19 loss-of-function (LOF) gene who take double doses of clopidogrel overcome the decreased efficacy of clopidogrel which partly due to CYP2C19 LOF gene, without increasing the risk of hemorrhage. (2) Individualized antiplatelet therapy based on CYP2C19 genotype and platelet function can significantly reduce the occurrence of MACE (mainly acute non-fatal myocardial infarction) after PCI without increasing the risk of moderate or severe hemorrhage.

[Investigation on Demodex infections among university students in Kunming City].

OBJECTIVE: To investigate the prevalence of Demodex infection among students in Kunming Medical University, and identify the factors affecting Demodex infections, so as to provide the evidence for the development of the strategy for the prevention of Demodex infections. METHODS: A total of 1 463 students from Grade 2014 who studied Medical Parasitology in Kunming Medical University were included in the survey. Demodex was examined in students'facial skin using the cellophane tape method, and the species was identified using microscopy. The students'gender, ethnicity, place of origin and skin type were captured using a questionnaire survey. RESULTS: The overall prevalence of Demodex infections was 19.07% (279/1 463) on the facial skin among the university students, and a higher prevalence was seen in girls (21.16%, 183/865) than in boys (16.05%, 96/598) (χ2 =5.965,P <0.05).TheprevalenceofDemodex infectionswas18.33%(66/360)amongminorethnicstudents,andnoethnicity-specific prevalence was seen (P > 0.05). Demodex folliculorum was the predominant species, with a prevalence of 50.54% (141/279), and mild infections were predominant among all infections (96.77%, 270/279), without severe infections seen. Multivariate nonconditional logistic regression analysis revealed that gender and roommates with Demodex infections were risk factors of Demodex infections, and the infection was not associated with ethnicity, place of origin or skin type. There were only 2.53% (37/1 463) of the subjects understanding the knowledge pertaining to the prevention and control of Demodex infection. CONCLUSIONS: A relatively low prevalence of Demodex infection is detected in the facial skin of students from Kunming Medical University, and Demodex infection is associated with gender and roommates with Demodex infections. Health education pertaining to the prevention of Demodex infections is suggested to be intensified among university students.

[Progress of researches on Fasciola and fascioliasis in Yunnan Province].

Fascioliasis is a zoonotic parasitic disease that seriously endangers human health and hinders socioeconomic development. Fasciola mainly infects ruminants, such as cattle and sheep. However, there has recently been a rise in the number of human cases with fascioliasis with the improvements of diagnostic techniques. During the past decades, sporadic cases of fascioliasis were predominantly identified; however, there were outbreaks of fascioliasis in Yunnan Province, which has been paid much attention. The review summarizes the advances in the distribution of Fasciola species and the progress of researches on fascioliasis in Yunnan Province.

Neuropilin-1 contributes to esophageal squamous cancer progression via promoting P65-dependent cell proliferation.

Neuropilin-1 (NRP1) is a non-kinase receptor recently implicated in tumor progression. Here we revealed that over-expression of NRP1 correlates with poor prognosis in esophageal squamous cell carcinoma (ESCC). NRP1-knockdown suppressed ESCC cell proliferation and xenograft tumor growth. Reduced NRP1 expression downregulated P65 mRNA and protein expression, and ectopic expression of P65-restored cell proliferation in NRP1-silenced cells. NRP1 regulates P65 transcription by activating cAMP responsive element binding protein (CREB). NRP1 interacted with and activated epidermal growth factor receptor (EGFR), and b1/b2 domain of NRP1 is responsible for the activation of EGFR. We also found that EGFR regulated CREB transcriptional activity via AKT. These data suggest that NRP1 is an upstream regulator in the P65-dependent proliferation signaling pathway and a candidate therapeutic target for ESCC.

Genome sequence compression based on optimized context weighting.

Context weighting is an important technology for genome compression. In this study, we discuss the relationship between the weighting of context models and the weighting of the description lengths corresponding to their respective context models. It indicates that weighting of context models is equivalent to the weighting of their description lengths. With these discussions, we present the weights optimization algorithm based on the minimum description length, and suggest implementing the least-square algorithm for the optimization of the weights. The proposed optimization algorithm is used in the compression of bacterial genome sequences. The experiment results indicate that by using the proposed weights optimization method, our context weighting-based genome compression algorithm can achieve better performance than context weighting-based algorithms reported in the literature.

[Expression and intratumoral heterogeneity of LN-5γ2 in esophageal squamous cell carcinoma].

OBJECTIVE: The present study aimed to evaluate the expression and intratumoral heterogeneity of LN-5γ2 in esophageal squamous cell carcinoma (ESCC). METHODS: The expression of LN-5γ2 protein was examined in 135 ESCC cases by immunohistochemistry, and to analyze its relationship with the clinical relevance of patients. The protein expressions in different regions in the same tumor as well as different nests in the same region were compared. RESULTS: Moderate and high expression of LN-5γ2 protein was detected in 40.0% (54/135) of tumor tissues. Positive immunohistochemical staining was observed in 31.1% (23/74) of early stage (stages Ⅰ/Ⅱ) cases and 50.8% (31/61) of late stage (stage Ⅲ) cases, with a significant difference between these two groups (P=0.023). There was no statistical association of LN-5γ2 expression with age, sex of patients, PT sage, lymph node metastasis and degree of tumor differentiation (P>0.05). However, differential expression of LN-5γ2 protein was found at different sampling sites in the same tumor and the same sampling site in different carcinomas. CONCLUSION: High expression of LN-5γ2 is positively correlated with tumor clinical stages and there existed intratumoral heterogeneity of LN-5γ2 expression in ESCC tissues.

Microtubule-associated protein 4 is an important regulator of cell invasion/migration and a potential therapeutic target in esophageal squamous cell carcinoma.

Cell invasion and migration significantly contribute to tumor metastasis. Microtubule-associated protein 4 (MAP4) protein is one member of microtubule-associate proteins family. It is responsible for stabilization of microtubules by modulation of microtubule dynamics. However, there is little information about the involvement of MAP4 in human cancer. Here we show that MAP4 serves as a regulator of invasion and migration in esophageal squamous cancer cells. By activating the ERK-c-Jun-vascular endothelial growth factor A signaling pathway, MAP4 promotes cell invasion and migration in vitro, tumor growth and metastasis in mouse models. Immunohistochemical staining of operative tissues indicated that MAP4 expression was associated with tumor stage, lymph node metastasis and shorter survival of the patients with esophageal squamous cell carcinoma (ESCC). Multivariate Cox regression analysis showed that MAP4 is an independent prognostic indicator. In the serial sections of ESCC tissues, there was a positive correlation between MAP4 and vascular endothelial growth factor A expression. Notably, an intratumoral injection of MAP4-small interfering RNA (siRNA) remarkably inhibited the growth of the tumors that formed by the MAP4-expressing ESCC cells in nude mice, and a combination of MAP4-siRNA and Bevacizumab significantly enhanced the inhibition effect. Our data suggest that MAP4 is probably a useful prognostic biomarker and a potential therapeutic target for the disease.

Efficacy and safety of phosphodieterase-5 inhibitors for treatment of erectile dysfunction secondary to spinal cord injury: a systemic review and meta-analysis.

STUDY DESIGN: Systemic reviewObjective:We carried out a systematic review and meta-analysis to assess the efficacy and safety of phosphodieterase-5 (PDE5) inhibitors on erectile dysfunction (ED) secondary to spinal cord injury (SCI). METHODS: A literature review was performed to identify all published randomized, double-blind, placebo-controlled trials of PDE5 inhibitors for treatment of ED secondary to SCI. The search included the following database: MEDLINE, EMBASE and the Cochrane Library. The outcomes and complications analyzed involved the Global Efficacy Question (GEQ), sexual encounter profile diary question 2 and 3 (SEP2 and SEP3) and adverse events. All statistical analysis was performed using Stata 12.0 software (Stata Corp., College Station, TX, USA). RESULTS: Six publications were used in analysis, including six randomized controlled trials that compared PDE5 inhibitors with placebo. Compared with placebo, PDE5 inhibitors were associated with significant improvements in GEQ (OR 11.997, 95% CI 8.073-17.830, P<0.0001), SEP2 (RR 1.847, 95% CI 1.561-2.185, P<0.0001) and SEP3 (RR 2.738, 95% CI 2.084-3.598, P<0.0001). Despite significant greater incidences of some adverse events observed (headache: RR 3.717, 95% CI 2.309-5.982, P<0.0001; flushing: RR 9.281, 95% CI 2.858-30.147, P<0.0001; gastrointestinal discomfort: RR 9.064, 95% CI 2.116-38.827, P=0.003), most adverse events were mild to moderate and transient. CONCLUSIONS: This systematic review and meta-analysis indicate that PDE5 inhibitors are effective and well tolerated to treat ED secondary to SCI compared with placebo, as measured by response to GEQ, SEP2, SEP3 and incidence of adverse events. PDE5 inhibitors could be considered as the first choice in the treatment of ED patients with SCI.

Screening of potential biomarkers for cholangiocarcinoma by integrated analysis of microarray data sets.

Cholangiocarcinoma (CCA) continues to harbor a difficult prognosis and it is difficult to diagnose in its early stages. The molecular mechanisms of CCA oncogenesis and progression are poorly understood. This study aimed to identify candidate biomarkers for CCA. Integrated analysis of microarray data sets was performed to identify differentially expressed genes (DEGs) between CCA and normal tissues. Gene Ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were then performed to identify the functions of DEGs. Furthermore, the protein-protein interaction (PPI) network of DEGs was constructed. The expressions of DEGs were validated in human CCA tissues by qRT-PCR. A set of 712 DEGs were identified in CCA compared with normal tissues, including 306 upregulated and 406 downregulated DEGs. It can be shown from the KEGG pathway analysis that some pathways may have important roles in pathology of CCA, including peroxisome proliferator-activated receptor signaling pathway, bile secretion, cell cycle, fat digestion and absorption. PPI network indicated that the significant hub proteins were PKM, SPP1 and TPM1. The abnormally overexpression PKM, SPP1 and TPM1 were closely related to oncogenesis and progression of CCA. PKM, SPP1, TPM1, COL1A1 and COL1A2 may serve as candidate biomarkers for diagnosis and prognosis of CCA.

HLA-B*13:01 and the dapsone hypersensitivity syndrome.

BACKGROUND: Dapsone is used in the treatment of infections and inflammatory diseases. The dapsone hypersensitivity syndrome, which is associated with a reported mortality of 9.9%, develops in about 0.5 to 3.6% of persons treated with the drug. Currently, no tests are available to predict the risk of the dapsone hypersensitivity syndrome. METHODS: We performed a genomewide association study involving 872 participants who had received dapsone as part of multidrug therapy for leprosy (39 participants with the dapsone hypersensitivity syndrome and 833 controls), using log-additive tests of single-nucleotide polymorphisms (SNPs) and imputed HLA molecules. For a replication analysis, we genotyped 24 SNPs in an additional 31 participants with the dapsone hypersensitivity syndrome and 1089 controls and performed next-generation sequencing for HLA-B and HLA-C typing at four-digit resolution in an independent series of 37 participants with the dapsone hypersensitivity syndrome and 201 controls. RESULTS: Genomewide association analysis showed that SNP rs2844573, located between the HLA-B and MICA loci, was significantly associated with the dapsone hypersensitivity syndrome among patients with leprosy (odds ratio, 6.18; P=3.84×10(-13)). HLA-B*13:01 was confirmed to be a risk factor for the dapsone hypersensitivity syndrome (odds ratio, 20.53; P=6.84×10(-25)). The presence of HLA-B*13:01 had a sensitivity of 85.5% and a specificity of 85.7% as a predictor of the dapsone hypersensitivity syndrome, and its absence was associated with a reduction in risk by a factor of 7 (from 1.4% to 0.2%). HLA-B*13:01 is present in about 2 to 20% of Chinese persons, 1.5% of Japanese persons, 1 to 12% of Indians, and 2 to 4% of Southeast Asians but is largely absent in Europeans and Africans. CONCLUSIONS: HLA-B*13:01 was associated with the development of the dapsone hypersensitivity syndrome among patients with leprosy. (Funded by the National Natural Science Foundation of China and others.).

Optimization of a bacterial consortium for nitrobenzene degradation.

Three bacterial strains, Arthrobacter sp. NB1, Serratia sp. NB2 and Stenotrophomonas sp. NB3, were isolated from contaminated sludge by using nitrobenzene as a sole source of carbon and nitrogen. It was observed that all three strains could degrade nitrobenzene at 400 mg/L initial concentration and mixed-cultivation of these strains could enhance the degradation of nitrobenzene compared with mono-cultivation. Mixture design was used for adjusting the proportions of each strain and the optimal ratio of inoculation size was NB1:NB2:NB3 = 4:4:5, where the nitrobenzene degradation percentage was two times higher than for by the single strain. The results of Plackett-Burman design indicated that Mg(2+), Ca(2+), Fe(2+), Zn(2+) and Mn(2+) had a positive effect on the degradation of nitrobenzene, while Cu(2+) and Co(2+) had a negative effect on it.

Effect of thyroxine on SNARE complex and synaptotagmin-1 expression in the prefrontal cortex of rats with adult-onset hypothyroidism.

Thyroid hormone insufficiency in adulthood causes a wide range of brain impairments, including altered synaptic proteins in the prefrontal cortex (PFC). The present study investigated whether adult-onset hypothyroidism altered the expression of soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes and synaptotagmin-1 (syt-1) in the PFC of rats. Sprague-Dawley rats were randomly divided into 4 groups: control, hypothyroid, and hypothyroid treated with T(4) [5 or 20 µg/100 g body weight (BW)]. Adult-onset hypothyroidism was induced in rats with the antithyroid drug 6-n-propyl-2-thiouracil (ip injection). PFC levels of synaptosomal-associated protein of 25 kDa (SNAP-25), syntaxin-1, vesicle-associated membrane protein 2 (VAMP-2) and syt-1 were determined by immunohistochemistry and western blot analyses. The results showed that syntaxin-1 and syt-1 were expressed at significantly lower levels in hypothyroid rats, VAMP-2 levels were not altered, and SNAP-25 levels were much higher compared to controls. A 2-week treatment with 5 µg T(4)/100 g BW partially normalized levels of SNARE complex and syt-1, and 20 µg T(4)/100 g BW restored these proteins closer to normal levels. Our findings indicate that dysregulation of SNARE complex and syt-1 in PFC of adult-onset hypothyroidism can be restored by T(4) treatment.

The decolorisation capacity and mechanism of Shewanella oneidensis MR-1 for methyl orange and acid yellow 199 under microaerophilic conditions.

Shewanella oneidensis MR-1 was found to reach 99.36% and 78.25% decolorisation for Methyl Orange and Acid Yellow 199 in solutions, respectively. The suitable pH range for decolorisation of Methyl Orange and Acid Yellow 199 by S. oneidensis MR-1 was 4.0-7.0 and 6.0-8.0, respectively, The azo dyes' removal by S. oneidensis MR-1 was slightly enhanced by addition of Mg(2+), but inhibited by Pb(2+), Cd(2+), Cu(2+), Fe(3+) and Fe(2+). The enzyme activities of NADH-DCIP reductase and azoreductase were 2.67 and 3.0 times higher, and 1.92 and 2.48 times higher, respectively, in the Methyl Orange treatment and in the Acid Yellow 199 treatment as compared to the control treatment. These findings indicated that the azo dyes' decolorisation by S. oneidensis MR-1 was via reduction mechanism.

Effect of thyroxine on synaptotagmin 1 and SNAP-25 expression in dorsal hippocampus of adult-onset hypothyroid rats.

Adult-onset hypothyroidism causes cognitive dysfunctions of learning and memory, in which many synaptic proteins in hippocampus are involved. In our work, we studied the effect of adult-onset hypothyroidism on the expression of synaptotagmin 1 (syt 1) and SNAP-25 in dorsal hippocampus as well as its recovery by levothyroxine (L-T(4)) replacement therapy. Rats were divided into 4 groups: control, hypothyroidism, and hypothyroid rats treated with 5 µg T(4)/100 g body weight (BW) and 20 µg L-T(4)/100 g BW, respectively. Protein levels of syt 1 and SNAP-25 in dorsal hippocampus were determined by Western blot and immunohistochemistry. The immunoblot analysis indicated that syt 1 was expressed at a significantly lower level in hypothyroid rats, while the level of SNAP-25 was much higher compared to controls. Furthermore, using immunostaining, we found that on the one hand, expression of syt 1 was significantly down-regulated in the examined layers of CA1 and CA3 subregions but not dentate gyrus (DG); however, on the other hand, expression of SNAP-25 was up-regulated in the layers of CA1, CA3, and DG. Two-week treatment with 20 µg LT(4)/ 100 g BW fully restored the levels of syt 1 and SNAP-25 to the normal level, which was more effective than 5 µg LT(4)/ 100 g BW that partially restored the levels of both proteins. These results suggest that adult-onset hypothyroidism caused down-regulation of syt 1 and up-regulation of SNAP- 25 level in dorsal hippocampus, which could be restored by L-T(4) treatment, and the recovery degree is related to the LT(4) dosage.

Sensitivity of pooled serum testing for screening antibody of schistosomiasis japonica by IHA in a mountainous area of Yunnan, China.

Pooled sample testing (PST) as a strategy for avoiding testing the majority of individual negative samples has been proposed for screening of diseases in low prevalence areas. There has been no standard guideline for PST in screening of Schistosoma japonicum infection of Yunnan, China. To document the optimum pool size with acceptable sensitivity of PST for screening of Schistosoma japonicum infection in this setting, an experimental pooling of each of 31 positive sera by IHA with various numbers of 24 negative sera was done. The results were used to create a statistical model which was subsequently used for simulation to predict sensitivity of the pooled serum tests in the population with varying prevalence and pool size. We found that to keep the sensitivity of PST above 90%, 1:05 should be the maximum dilution, that is, the optimum pool size should not be greater than 6. Antigen will have rather little interference if the prevalence of infection is low e.g. 1% or the antigen:antibody ratio is 1:100 or below. Pooled serum testing by IHA is an acceptable sensitive method for detecting antibody for Schistosoma japonicum infection in this area.

RTA2 is involved in calcineurin-mediated azole resistance and sphingoid long-chain base release in Candida albicans.

The calcineurin pathway has been reported to be essential for the development of azole resistance in Candida albicans. The depletion or ectopic over-expression of RTA2 increased or decreased susceptibility of C. albicans to azoles, respectively. CaCl(2)- induced activation of the calcineurin pathway in wildtype C. albicans promoted resistance to azoles, while the Ca(2+) chelator (EGTA), calcineurin inhibitors (FK506 and cyclosporin A) and the deletion of RTA2 blocked the resistance-promoting effects of CaCl(2). Furthermore, we found that RTA2 was up-regulated in a calcineurin-dependent manner. The depletion of RTA2 also made the cell membrane of C. albicans liable to be destroyed by azoles and RTA2 over-expression attenuated the destroying effects. Finally, the disruption of RTA2 caused an increased accumulation of dihydrosphingosine (DHS), one of the two sphingolipid long-chain bases, by decreasing release of DHS. In conclusion, our findings suggest that RTA2 is involved in calcineurin-mediated azole resistance and sphingoid long-chain base release in C. albicans.

Isolation of a homocysteine gamma-lyase-producing bacterium and study of its enzyme production conditions.

AIM: To investigate the possibility of finding a new homocysteine (Hcy) gamma-lyase with the desired properties for Hcy measurement in bacteria. METHODS AND RESULTS: Through a process of enrichment, the Hcy gamma-lyase-producing bacterium strain N2-1 was isolated from soil. Based upon its morphological, physiological, and biochemical characteristics, as well as its 16S rDNA sequence and phylogenetic tree analysis, this isolate belongs to the genus Serratia. The effects of pH, aeration, inducers, carbon (C) and nitrogen (N) sources on enzyme production were studied. Methionine, yeast extract, and glucose were selected as the optimal inducer, C and N sources, respectively. Maximum production of Hcy gamma-lyase was obtained when the isolate was cultured at 30 degrees C at pH 6.5 for about 36 h in the optimum medium. Results also showed that this Hcy gamma-lyase has relatively high specificity towards Hcy. CONCLUSIONS: Because of its high specificity for Hcy, this bacterial Hcy gamma-lyase has the potential application in Hcy determination. SIGNIFICANCE AND IMPACT OF THE STUDY: In addition to isolating a bacterium that produces Hcy gamma-lyase suitable for Hcy determination, this study also indicates that the bacterium could be a source for production of Hcy gamma-lyase for clinical applications.