Comparison of clinical features of patients with or without severe gastrointestinal complications in aggressive gastrointestinal lymphomas

BACKGROUND Aggressive primary gastrointestinal non-Hodgkin lymphoma(PGINHL)is an uncommon and heterogeneous group of lymphoid malignancies,that differs from indolent lymphoma and has a high incidence of severe gastrointestinal complications(GICs).AIM To investigate and compare the clinicopathological characteristics,treatments and outcomes in the GICs and No-GICs group with aggressive PGINHL.METHODS This retrospective analysis was performed on aggressive PGINHL patients between January 2013 and December 2021 at our hospital.The independent influence factors of GICs were obtained by univariate and multivariate Logistic regression analysis,the selected variables significantly related to GICs were selected as the final predictors to construct nomogram.Kaplan-Meier curves further analyzed the survival of patients in GICs and No-GICs groups.Survival analysis of GICs group was performed using Cox regression.RESULTS We focused on 124 aggressive PGINHL cases,which had a relatively high incidence 48.4%(60/124 cases)of GICs,the most common histological type in GICs group was diffuse large B-cell lymphoma(DLBCL)(n=49,81.7%).In the GICs group,small intestine was the most common anatomic site of lesion(43.3%),followed by large intestine(31.7%),and then stomach and esophagus(25.0%).Multivariate Logistic regression analysis showed that the independent risk factors for GICs were the small intestine[odd ratio(OR)=3.33;95%confidence interval(CI):1.47-9.41;P=0.009),aggressive B-cell(OR=0.09;95%CI:0.01-0.83;P=0.033),maximum tumor diameter(OR=1.25;95%CI:1.07-1.47;P=0.005),invaded deep serous layer(OR=3.38;95%CI:1.24-9.19;P=0.017).We developed a nomogram to predict risk of GICs in aggressive PGINHL patients based on independent risk factors.The value of area under curve calculated by receiver operating characteristic curve was 0.815,and calibration curve and decision curve analysis further indicated that the prediction effect was superior.The majority of patients with GICs were given combination therapy(chemotherapy combined with surgery or radiation).Event-free survival and overall survival in GICs group were no worse than those in the No-GICs group.CONCLUSION The complication rate of GICs in patients with aggressive PGINHL was relatively high,particularly in PGI-DLBCL.The independent risk factors for GICs were the small intestine,PGI-TNKL,bulky tumor,and depth of invasion.A combination treatment,involving surgery,improved survival in the GICs group.

Value of improved nursing measures and enhanced nursing management to reduce the occurrence of adverse events in pediatric infusion

BACKGROUND Intravenous infusion is a common method of drug administration in clinical practice.Errors in any aspect of the infusion process,from the verification of medical orders,preparation of the drug solution,to infusion by nursing staff,may cause adverse infusion events.AIM To analyzed the value of improving nursing measures and enhancing nursing management to reduce the occurrence of adverse events in pediatric infusion.METHODS The clinical data of 130 children who received an infusion in the pediatric department of our hospital from May 2020 to May 2021 were analyzed and divided into two groups according to the differences in nursing measures and nursing management:65 patients in the control group received conventional nursing and nursing management interventions,while 65 patients in the observation group received improved nursing measure interventions and enhanced nursing management.The occurrence of adverse events,compliance of children,satisfaction of children’s families,and complaints regarding the transfusion treatment were recorded in both groups.RESULTS The incidence of fluid extravasation and infusion set dislodgement in the observation group were 3.08%and 1.54%,respectively,which were significantly lower than 12.31%and 13.85%in the control group(P<0.05),while repeated punctures and medication addition errors in the observation group were 3.08%and 0.00%,respectively,which were lower than 9.23%and 3.08%in the control group,but there was no significant difference(P>0.05).The compliance rate of children in the observation group was 98.46%(64/65),which was significantly higher than 87.69%(57/65)in the control group,and the satisfaction rate of children’s families was 96.92%(63/65),which was significantly higher than 86.15%(56/65)in the control group(P<0.05).The observation group did not receive any complaints from the child’s family,whereas the control group received four complaints,two of which were due to the crying of the child caused by repeated punctures,one due to the poor attitude of the nurse,and one due to medication addition errors,with a cumulative complaint rate of 6.15%.The cumulative complaint rate of the observation group was significantly lower than that of the control group(P<0.05).CONCLUSION Improving nursing measures and enhancing nursing management can reduce the incidence of fluid extravasation and infusion set dislodgement in pediatric patients,improve children’s compliance and satisfaction of their families,and reduce family complaints.

Interoception mediates the association between social support and sociability in patients with major depressive disorder

BACKGROUND Interoception dysfunction has an important impact on the onset and development of major depressive disorder(MDD).Social support serves as a protective factor against MDD,and sociability also plays a significant role in this condition.These interconnected constructs-social support and sociability-play pivotal roles in MDD.However,no research on the mechanisms underlying the associations be-tween social support and sociability,particularly the potential role of interocep-tion,have been reported.ception,social support,and sociability,respectively.A mediation analysis model for the eight dimensions of intero-ception(noticing,not distracting,not worrying,attention regulation,emotional awareness,self-regulation,body listening,and trust),social support,and sociability were established to evaluate the mediating effects.RESULTS A partial correlation analysis of eight dimensions of the MAIA-2,SSRS,and TSBI scores,with demographic data as control variables,revealed pairwise correlations between the SSRS score and both the MAIA-2 score and TSBI score.In the major depression(MD)group,the SSRS score had a positive direct effect on the TSBI score,while the scores for body listening,emotional awareness,self-regulation,and trust in the MAIA-2C had indirect effects on the TSBI score.In the HC group,the SSRS score had a positive direct effect on the TSBI score,and the scores for attention regulation,emotional awareness,self-regulation,and trust in the MAIA-2C had indirect effects on the TSBI score.The proportion of mediators in the MD group was lower than that in the HC group.CONCLUSION Interoceptive awareness is a mediating factor in the association between social support and sociability in both HCs and depressed patients.Training in interoceptive awareness might not only help improve emotional regulation in depressed patients but also enhance their social skills and support networks.

Molecular mechanism of hepatitis B virus X protein function in hepatocarcinogenesis

Many factors are considered to contribute to hepatitis B virus(HBV)-associated hepatocellular carcinoma(HCC),including products of HBV,HBV integration and mutation,and host susceptibility. HBV X protein(HBx) can interfere with several signaling pathways associated with cell proliferation and invasion,and HBx C-terminal truncation has been suggested to impact the development of HCC. This review focuses on the pathological functions of HBx in HBV-induced hepatocarcinogenesis. As a transactivator,HBx can affect regulatory non-coding RNAs(nc RNAs),including micro RNAs and long nc RNAs. HBx is also involved in epigenetic modification and DNA repair. HBx interacts with various signal-transduction pathways,such as the p53,Wnt,and nuclear factor-κB pathways. We conclude that HBx hastens the development of hepatoma.

COOH-terminal deletion of HBx gene is a frequent event in HBV-associated hepatocellular carcinoma

AIM:To investigate the hepatitis B virus (HBV) x gene (HBx) state in the tissues of HBV-related hepatocellular carcinoma (HCC) in Chinese patients and whether there were particular HBx mutations. METHODS: HBx gene was amplified and direct sequencing was used in genomic DNA samples from 20 HCC and corresponding non-cancerous liver tissues from HBsAg-positive patients. HBV DNA integration and HBx deleted mutation were validated in 45 HCC patients at different stages by Southern blot analysis and polymerase chain reaction methods. RESULTS: The frequencies of HBx point mutations were significantly lower in HCC than their corresponding non- cancerous liver tissues (11/19 vs 18/19, P = 0.019). In contrast, deletions in HBx gene were significantly higher in HCC than their non-cancerous liver tissues (16/19 vs 4/19, P < 0.001). The deletion of HBx COOH-terminal was detected in 14 HCC tissues. A specific integration of HBx at 17p13 locus was also found in 8 of 16 HCC, and all of them also exhibited full-length HBx deletions. Integrated or integrated coexistence with replicated pattern was obtained in 45.5% (20/45) - 56.8% (25/45) tumors and 40.9% (18/45) - 52.3% (23/45) non-tumor tissues. CONCLUSION: HBx deletion, especially the COOH- terminal deletion of HBx is a frequent event in HBV-associated HCC tissues in China. HBV integration had also taken place in partial HCC tissues. This supporting the hypothesis that deletion and probably integrated forms of the HBx gene may be implicated in liver carcinogenesis.

Is frailty associated with short-term outcomes for elderly patients with acute coronary syndrome？

Background Frailty is a new prognostic factor in cardiovascular medicine due to the aging and increasingly complex nature of elderly patients. It is useful and meaningful to prospectively analyze the manner in which frailty predicts short-term outcomes for elderly patients with acute coronary syndrome （ACS）. Methods Patients aged 〉 65 years, with diagnosis of ACS from cardiology department and geriatrics department were included from single-center. Clinical data including geriatrics syndromes were collected using Comprehensive Geriatrics Assessment. Frailty was defined according to the Clinical Frailty Scale and the impact of the co-morbidities on risk was quantified by the coronary artery disease （CAD）--specific index. Patients were followed up by clinical visit or telephone consultation and the median follow-up time is 120 days. Following-up items included all-cause mortality, unscheduled return visit, in-hospital and recurrent major adverse cardiovascular events. Multivariable regression survival analysis was performed using Cox regression. Results Of the 352 patients, 152 （43.18%） were considered frail according to the study instrument （5-7 on the scale）, and 93 （26.42%） were considered moderately or se- verely frail （6-7 on the scale）. Geriatrics syndromes including incontinence, fall history, visual impairment, hearing impairment, constipation, chronic pain, sleeping disorder, dental problems, anxiety or depression, and delirium were more frequently in frail patients than in non-frail patients （P = 0.000, 0.031, 0.009, 0.014, 0.000, 0.003, 0.022, 0.000, 0.074, and 0.432, respectively）. Adjusted for sex, age, severity of coro- nary artery diseases （left main coronary artery lesion or not） and co-morbidities （CAD specific index） by Cox survival analysis, frailty was found to be strongly and independently associated with risk for the primary composite outcomes： all-canse mortality [Hazard Ratio （HR） = 5.393; 95% CI： 1.477-19.692, P = 0.011] and unscheduled return visit （HR - 2.832; 95% CI： 1.140-7.037, P = 0.025）. Conclusions Comprehensive Geriatrics Assessment and Clinical Frail Scale were useful in evaluation of elderly patients with ACS. Frailty was strongly and independently associated with short-term outcomes for elderly patients with ACS.

Loss of Wnt5a and Ror2 protein in hepatocellular carcinoma associated with poor prognosis

AIM： To investigate the expression and clinical significance of Wnt member 5a （Wnt5a） and receptor tyrosine kinase-like orphan receptor 2 （Ror2） in hepatocellular carcinoma （HCC）.METHODS： In HCC tissues obtained from 85 patients, the protein expressions of Wnt5a, Rot2, 13-catenin, and Ki-67 via immunohistochemical staining using the Envision Plus System. The antibody binding was visualized with 3, 3＇-diaminobenzidine tetrahydrochloride （DAB） before brief counterstaining with Mayer＇s hematoxylin. The degree of immunohistochemical staining was recorded using a semiquantitative and subjective grading system. The mRNA expression of Rot2 was examined by real time reverse transcription polymerase chain reaction, including nineteen of the 85 HCC and three normal liver tissues. The ratios of Ror2 to the housekeeping gene GAPDH represented the normalized relative levels of Ror2 expression. To determine the prognostic factor, the outcome of the 82 patients was determined by reviewing their medical charts. The overall and diseasefree survival rates were estimated using the Kaplan-Meier method and compared with the logrank test. The prognostic analysis was carried out with univariate and multivariate Cox regressions models.RESULTS： Compared to nontumorous （hepatitis or cirrhotic） tissues, Rot2 mRNA expression was clearly decreased in HCC. Rot2 and Wnt5a protein expressions in the majority of HCC patients （63% and 77%, respectively） was significantly less in tumor tissues, as compared to adjacent nontumorous tissues, and this reduction was correlated with increasing serum α-fetoprotein and tumor stage. In 68% （58/85） of the HCC cases, the expression of IB-catenin in tumor tissues was either downregulated in the cellular membrane, upregulated in the cytoplasm, or both. Survival analysis indicated that Wnt5a and Ror2 protein expressions could be regarded as independent prognostic factors for HCC; HCC patients with decreased Wnt5a or Ror2 protein expression had a poorer prognosis than those with elevated Wnt5a and Ror2 expression （P = 0.016, P = 0.007, respectively）.CONCLUSION： Wnt5a and Ror2 may serve as tumor suppressor genes in the development of HCC, and may serve as clinicopathologic biomarkers for prognosis in HCC patients.

Prevalence and prognostic implications of sarcopenia in older patients with coronary heart disease

Background Sarcopenia is a progressive and generalized skeletal muscle disorder that is associated with an increased likelihood of adverse outcomes, including falls, fractures, physical disability, and mortality. However, there have been few systematic studies of the prevalence and prognostic values of sarcopenia in older patients with coronary heart disease(CHD). This study aimed to investigate the prevalence of sarcopenia in hospitalized older patients with CHD, and to prospectively evaluate the effect of sarcopenia on the short-term prognosis of these patients. Methods Patients aged ≥ 65 years, with the diagnosis of CHD from Peking Union Medical College Hospital between December 2017 and November 2018, were included. Sarcopenia was diagnosed according to consensus of the Asian Working Group for Sarcopenia in 2014. Follow-up items included unscheduled return visits, occurrence of major adverse cardiac and cerebral events (MACCE), and all-cause mortality. The MACCE-free survival curve of sarcopenic and non-sarcopenic older patients with CHD was estimated by the Kaplan-Meier method. Cox regression analysis was used to analyze the association between sarcopenia and an unscheduled return visits, MACCE, and all-cause mortality. Results A total of 345 older patients with CHD were enrolled in the study, with a median age of 74 years. Among the patients, 78 (22.6%) were diagnosed with sarcopenia. During the follow-up time, there were significantly more unscheduled return visits in sarcopenic patients than in non-sarcopenic patients (34.2% vs. 21.8%,χ^2 = 4.418, P = 0.036), while there was no significant difference in the occurrence of MACCE (χ^2 = 2.869, P = 0.09) or all-cause mortality (χ^2 = 1.673, P = 0.196) between these patient groups. The Kaplan-Meier curve showed that the MACCE-free survival time of sarcopenic patients was significantly shorter than that in non-sarcopenic patients (χ^2 = 4.102, P = 0.043). After adjusting for sex, age, and the Charlson comorbidity index, sarcopenia was not an independent risk factor of unscheduled return visits (HR = 1.002, 95% CI: 0.556–1.807). However, the complication of anxiety and depression was an independent risk factor (HR = 1.876, 95% CI: 1.012–3.477, P = 0.046) for unscheduled return visits in older patients with CHD. Conclusions There is a high prevalence of sarcopenia among hospitalized older adults with CHD. A shorter MACCE-free survival time and more unscheduled return visits are found in sarcopenic older patients with CHD. Clinicians should pay more attention to the functional status of older patients with CHD, as well as identification and management of geriatric syndromes.

Measurement of professional identity in Chinese nursing students

Objectives:Chinese nurses continue to display low professional identity.It is becoming an emergent issue in China how to help nurse students develop their positive professional identity.This paper is to develop the Professional Identity Scale for Nursing Students(PISNS).Methods:Literature review,and interviews with students and experts were adopted to develop initial item.Reliability and validity of the scale were respectively examined by computing internal consistency coefficient alpha and split-half Spearman Brown for equal length,and Exploratory Principle Component Factor Analysis.Results:Five factors with 17-item were obtained explaining 58.9%of the total variance.Cronbach’s alpha,and split-half reliability was 0.83 and 0.84 respectively.Conclusions:Given its high reliability and validity,PISNS could be used as a measurement tool for educators to evaluate the developing level of professional identity in nursing students and assess the effectiveness of corresponding interventions and strategies.

Fluorescent co-localization of PTS1 and PTS2 and its application in analysis of the gene function and the peroxisomal dynamic in Magnaporthe oryzae

The peroxisomal matrix proteins involved in many important biological metabolism pathways in eukaryotic cells are encoded by nucleal genes, synthesized in the cytoplasm and then transported into the organelles. Targeting and import of these proteins depend on their two peroxisomal targeting signals （PTS 1 and PTS2） in sequence as we have known so far. The vectors of the fluorescent fusions with PTS, i.e., green fluorescence protein （GFP）-PTS1, GFP-PTS2 and red fluorescence protein （RFP）-PTS1, were constructed and introduced into Magnaporthe oryzae Guy ll cells. Transformants containing these fusions emitted fluorescence in a punctate pattern, and the locations of the red and green fluorescence overlapped exactly in RFP-PTS 1 and GFP-PTS2 co-transformed strains. These data indicated that both PTS1 and PTS2 fusions were imported into peroxisomes. A probable higher efficiency of PTS1 machinery was revealed by comparing the fluorescence backgrotmds in GFP-PTS1 and GFP-PTS2 transformants. By introducing both RFP-PTS1 and GFP-PTS2 into Amgpex6 mutants, the involvement of MGPEX6 gene in both PTS1 and PTS2 pathways was proved. In addition, using these transformants, the inducement ofperoxisomes and the dynamic of peroxisomal number during the pre-penetration processes were investigated as well. In summary, by the localization and co-localization of PTS1 and PTS2, we provided a useful tool to evaluate the biological roles of the peroxisomes and the related genes.

A novel gain of function mutant in C-kit gene and its tumorigenesis in nude mice

AIM： To transfect mutant C-kit cDNA at codon 579 into human embryonic kidney cell line to observe its role in the pathogenesis of gastrointestinal stromal tumor （GIST）. METHODS： Eukaryotic expression vectors of pcDNA3- Kit-NW and pcDNA3-Kit-W were constructed. Then pcDNA3-Kit-NW and pcDNA3-Kit-W plasrnids were transfected into human embryonic kidney cell line by Upofectamine. The resistant clone was screened by G418 filtration and identified by sequencing, Western blotting, and immunocytochemical staining. Human embryonic kidney cells were divided into three groups including pcDNA3-Kit-NW, pcDNA3-Kit-W, and vector control groups. Absorbency value with a wavelength of 574 nm was detected by MTT analysis. Mice were injected with three groups of cells. Volume, mass, and histological examinations of the tumors in different groups were measured and compared. RESULTS： The C-kit gene and mutant C-kit gene were successfully cloned into the eukaryotic expression vector pcDNA3, pcDNA3-Kit-NW and pcDNA3-Kit-W were successfully transfected into human embryonic kidney cell line and showed stable expression in this cell line. Cell proliferating activity had significant differences between pcDNA3-Kit-NW and pcDNA3, pcDNA3-Kit- NW and pcDNA3-Kit-W （P〈0.05）, respectively. Tumors were only observed in nude mice implanted with cells transfected with pcDNA3-Kit-NW. CONCLUSION： Mutation of C-kit gene increases the proliferation activity of human cells and plays an important role in the malignant transformation of GIST.

Characterization of 12 polymorphic microsatellite markers in the Chinese tree shrew (Tupaia belangeri chinensis)

The Chinese tree shrew (Tupaia belangeri chinensis) is a small experimental animal with a close affinity to primates. This species has long been proposed to be an alternative experimental animal to primates in biomedical research. Despite decades of study, there is no pure breed for this animal, and the overall genetic diversity of wild tree shrews remains largely unknown. In order to obtain a set of genetic markers for evaluating the genetic diversity of tree shrew wild populations and tracing the lineages in inbreeding populations, we developed 12 polymorphic microsatellite markers from the genomic DNA of the tree shrew. An analysis of a wild population of 117 individuals collected from the suburb of Kunming, China, showed that these loci exhibited a highly expected heterozygosity (0.616). These 12 microsatellites were sufficient for individual identification and parentage analysis. The microsatellite markers developed in this study will be of use in evaluating genetic diversity and lineage tracing for the tree shrew.

Angiopoietin-1 targeted RNA interference suppresses angiogenesis and tumor growth of esophageal cancer

AIM: To determine the inhibitory effect of the adenovirus- based angiopoietin-1 (Ang-1) targeted small interfering RNA expression system (Ad/Ang-1si) on the expression of the Ang-1 gene, cell growth and apoptosis in human esophageal cancer cell line Eca109. METHODS: siRNA-expressing adenovirus targeting Ang-1 gene was constructed using the Ad Easy System. Cultured Eca109 cells were transfected with Ad/Ang-1si (Eca109/Ang-1si), and Ad/si was used to infect Eca109 cells as control (Eca109/si). Ang-1 gene expression and concentration was determined with RT-PCR and ELISA, respectively. Human umbilical vein endothelial cell (HUVEC) migration and proliferation were analyzed. After s.c. injection into athymic nu/nu mice, the tumor growth, vessel density and apoptosis of each group was also determined. RESULTS: HUVEC migration induced by conditioned medium from Ang-1si-transfected Eca109 cells was significantly less than that induced by conditioned medium from Eca109 cells and control adenovirus- transfected Eca109 cells. Furthermore, after s.c. injection into athymic nu/nu mice, the tumor growth and cell apoptosis of Ad/Ang-1si -expressing Eca109 cells was significantly lower than that of parental or control adenovirus-transfected cells. Vessel density assessed by CD31 immunohistochemical analysis and Ang-1 expression by RT-PCR were also decreased. CONCLUSION: The targeting Ang-1 may provide a therapeutic option for esophageal cancer.

Fruit and vegetable consumption and its relation to risk of asthenopia among Chinese college students

AIM： To investigate the associations between fruit and vegetable consumption and risk of asthenopia among Chinese college students. METHODS： A total of 1022 students were selected from five universities by a multi-stage stratified cluster sampling method. They were surveyed via a self-administered questionnaire including socio-demographic features, dietary and lifestyle habits, eye-related symptoms, eye care habits and history of diseases. Ascertainment of asthenopia was based on participants＇ subjectively reported symptoms. The associations between fruit and vegetable intake with asthenopia risk were assessed using multivariate logistic regression analysis. RESULTS： There were no significant associations between total fruit and vegetable, total vegetable, or fruit and the risk of asthenopia. Higher intake of dark-green leafy vegetable was likely to be inversely associated with asthenopia risk [odd ratio （OR）： 0.60; 95%CI： 0.37-0.97; Ptrend=0.21] after controlling for nondietary and dietary risk factors. Stratified analysis showed that the inverse association between dark-green leafy vegetable intake and asthenopia risk was limited to participants with suboptimal eyesight （OR： 0.45; 95%CI： 0.25-0.82; Ptrend=0.05）, wearing glasses （OR： 0.35; 95%CI： 0.17-0.72; Ptrend=0.03） or using computer ≥3h/d （OR： 0.48; 95%CI： 0.25-0.93; Ptrend=0.08）. CONCLUSION： A higher consumption of dark-green leafy vegetable is associated with a lower asthenopia risk among college students with suboptimal eyesight and poor eye care habits.

Ahmed valve implantation for neovascular glaucoma after 23-gauge vitrectomy in eyes with proliferative diabetic retinopathy

·AIM: To report on the outcome of Ahmed glaucoma valve (AGV) implantation for the management of neovascular glaucoma (NVG) after 23 -gauge vitrectomy for proliferative diabetic retinopathy (PDR). ·METHODS: Twelve medically uncontrolled NVG with earlier 23 -gauge vitrectomy for PDR underwent AGV implantation. The control of intraocular pressure (IOP), preoperative and postoperative best -corrected visual acuity, the development of intraoperative and postoperative complications were evaluated during the follow-up. ·RESULTS: The mean follow-up was 15.4±4.3 months (9-23 months). Mean preoperative IOP was 49.4±5.1mmHg and mean postoperative IOP at the last visit was 17.5 ± 1.6mmHg. The control of IOP was achieved at the final follow -up visits in all patients, however, 8 of 12 patients still needed anti-glaucoma medication (mean number of medications, 0.8±0.7). The visual acuity improved in nine eyes, and the visual acuity unchanged in three eyes at the final follow -up visits. The complications that occurred were minor hyphema in three eyes, choroid detachment in two eyes, and the minor hyphema and choroid detachments were reabsorbed without any surgical intervention. ·CONCLUSION: AGV implantation is a safe and effective procedure that enables successful IOP control and vision preservation in the NVG patients with the history of earlier 23-gauge vitrectomy for PDR.·

Characterization of 12 polymorphic microsatellite markers in the Chinese tree shrew(Tupaia belangeri chinensis)

The Chinese tree shrew(Tupaia belangeri chinensis)is a small experimental animal with a close affinity to primates.This species has long been proposed to be an alternative experimental animal to primates in biomedical research.Despite decades of study,there is no pure breed for this animal,and the overall genetic diversity of wild tree shrews remains largely unknown.In order to obtain a set of genetic markers for evaluating the genetic diversity of tree shrew wild populations and tracing the lineages in inbreeding populations,we developed 12 polymorphic microsatellite markers from the genomic DNA of the tree shrew.An analysis of a wild population of 117 individuals collected from the suburb of Kunming,China,showed that these loci exhibited a highly expected heterozygosity(0.616).These 12 microsatellites were sufficient for individual identification and parentage analysis.The microsatellite markers developed in this study will be of use in evaluating genetic diversity and lineage tracing for the tree shrew.

A simple and effective method for total RNA isolation of appressoria in Magnaporthe oryzae

Appressorium formation is an important event in establishing a successful interaction between the rice blast fungus, Magnaporthe oryzae, and its host plant, rice. An understanding of molecular events occurring in appressorium differentiation will give new strategies to control rice blast. A quick and reliable method to extract total RNA from appressorium is essential for studying gene expression during appressorium formation and its mechanism. We found that duplicate film is an efficient substratum for appressorium formation, even when inoculated with high density conidia. When inoculated with conidia at 1 × 106 ml^-1, the percentages of conidium germination and appressorium formation were （97.98±0.67）% and （97.88±0.45）%, respectively. We applied Trizol before appressorium collection for total RNA isolation, and as much as 113.6 lag total RNA was isolated from the mature appressoria at 24 h after inoculation. Functional analysis of two genes, MNH6 and MgATG1, isolated from the cDNA subtractive library, revealed that the quantity of RNA was good enough to construct a cDNA （complementary DNA） library or a cDNA subtractive library. This method may be also applicable for the appressorium RNA isolation of other pathogenic fungi in which conidia differentiate into appressoria in the early stages of host infection.

Magnaporthe oryzae MTP1 gene encodes a type Ⅲ transmembrane protein involved in conidiation and conidial germination

In this study the MTP1 gene, encoding a type III integral transmembrane protein, was isolated from the rice blast fungus Magnaporthe oryzae. The Mtp 1 protein is 520 amino acids long and is comparable to the Ytp 1 protein of Saccharomyces cerevisiae with 46% sequence similarity. Prediction programs and MTP1-GFP （green fluorescent protein） fusion expression results indicate that Mtp 1 is a protein located at several membranes in the cytoplasm. The functions of the MTP1 gene in the growth and development of the fungus were studied using an MTP1 gene knockout mutant. The MTP1 gene was primarily expressed at the hyphal and conidial stages and is necessary for conidiation and conidial germination, but is not required for pathogenicity. The Amtpl mutant grew more efficiently than the wild type strain on non-fermentable carbon sources, implying that the MTP1 gene has a unique role in respiratory growth and carbon source use.

Controllable fabrication of self-organized nano-multilayers in copper–carbon films

In order to clarify the influence of methane concentration and deposition time on self-organized nano-multilayers,three serial copper-carbon films have been prepared at various methane concentrations with different deposition times using a facile magnetron sputtering deposition system. The ratios of methane concentration(CH4/Ar+CH4) used in the experiments are 20%, 40%, and 60%, and the deposition times are 5 minutes, 20 minutes, and 40 minutes, respectively.Despite the difference in the growth conditions, self-organizing multilayered copper-carbon films are prepared at different deposition times by changing methane concentration. The film composition and microstructure are investigated by x-ray photoelectron spectroscopy(XPS), x-ray diffraction(XRD), field emission scanning electron microscopy(FESEM), and high-resolution transmission electron microscopy(HRTEM). By comparing the composition and microstructure of three serial films, the optimal growth conditions and compositions for self-organizing nano-multilayers in copper-carbon film are acquired. The results demonstrate that the self-organized nano-multilayered structure prefers to form in two conditions during the deposition process. One is that the methane should be curbed at low concentration for long deposition time,and the other condition is that the methane should be controlled at high concentration for short deposition time. In particular, nano-multilayered structure is self-organized in the copper-carbon film with copper concentration of 10-25 at.%.Furthermore, an interesting microstructure transition phenomenon is observed in copper-carbon films, that is, the nanomultilayered structure is gradually replaced by a nano-composite structure with deposition time and finally covered by amorphous carbon.

Identification of tRNA-derived Fragments and Their Potential Roles in Atherosclerosis

Objective Atherosclerosis(AS),a chronic inflammatory disease,is the basis of cardiovascular disease(CVD).Although the treatment has been greatly improved,AS still imposes a large burden on human health and the medical system,and we still need to further study its pathogenesis.As a novel biomolecule,transfer RNA-derived fragments(tRFs)play a key role in the progression of various disease.However,whether tRFs contribute to atherosclerosis pathogenesis remains unexplored.Methods With deep sequencing technology,the change of tRFs expression profiles in patients with AS compared to healthy control group was identified.The accuracy of the sequencing data was validated using RT qPCR.Subsequently,we predicted the potential target genes of tRFs by online miRNA target prediction algorithms.The potential functions of tRFs were evaluated with Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses.Results There were 13 tRFs differentially expressed between patients with AS and healthy controls,of which 2 were up-regulated and 11 were down-regulated.Validation by RT-qPCR analysis confirmed the sequencing results,and tRF-Gly-GCC-009 was highly up-regulated in the AS group based on the results of sequencing which was confirmed by RT-qPCR analysis.Furthermore,GO enrichment and KEGG pathway analyses indicated that 10 signaling pathways were related to tRF-Gly-GCC-009.These pathways might be physiopathological fundamentals of AS,mainly involving in Apelin signaling,Notch signaling and calcium signaling.Conclusion The results of our study provide important novel insight into the underlying pathogenesis and demonstrate that tRFs might be potential biomarkers and therapeutic targets for AS in the future.