RNA-seq can help us quickly obtain the whole transcriptome sequences of species under different conditions. Many Unigenes that are assembled by raw reads always do not contain complete open reading frame (ORF). In addition, it also has some redundancy in transcriptome library. Some Unigenes in the library, although belong to one transcript, cannot be assembled without overlapping. We found five incomplete Unigenes annotated ammonium transporter (AMT) from Salicornia europaea transcriptome, in which two Unigenes (Uni4 and Uni5) had identical expression patterns across four transcriptomes. The two Unigenes may come from one transcript. Analyzing the Unigene position of transcript by NCBI blastx, we found that Uni4 and Uni5 respectively located in 5' end and 3' end compared with the reference transcript, and an unknown gap of 120 bp may exist in a hypothetic transcript to which Uni4 and Uni5 both belong. To verify the hypothesis, single forward primer and single reverse primers were respectively designed on Uni4 and Uni5, and a fragment with about 800 bp was generated by PCR. Then it was sequenced and aligned with Uni4 and Uni5. Finally, we assembled a sequence with 1 667 bp, which contains a complete ORF (1 482 bp, coding 494 amino acids). It belongs to amt1 subfamily and was named Seamt1 via the phylogenetic analysis. It was pointed by bioinformatics tools that SeAMT1 protein conformed to the AMT characteristics of other species. This work clustered expression pattern to explore the Unigenes of one transcript, and the feasibility of this method was validated through the other two groups of Unigenes. The handy method will benefit extension and assembling of Unigene in transcriptome, it also helps achieve the complete ORF and gene function.
Ammonium Compounds ; Chenopodiaceae ; genetics ; Computational Biology ; Gene Expression Profiling ; Genes, Plant ; Membrane Transport Proteins ; genetics ; Molecular Sequence Annotation ; Open Reading Frames ; Phylogeny ; Plant Proteins ; genetics ; Transcriptome

Objective:To prepare Dermatophagoides farinae (Der f) crude protein to establish BALB/c bronchial asthma model , and to observe the morphology and degranulation of mast cells and detect related cytokines .Methods: Dermatophagoides farinae ( Der f) crude protein were prepared by trituration .30 BALB/c mice were randomly divided into 3 groups:PBS control group (A), asthma model group (B) and Der f crude protein treatment group (C).Group A were treated with PBS(100 μl) all the time, group B and group C were treated with 50 μg Der f crude protein mixed with 50μl alum adjuvant on day 0,day 7 and day 14.On day 28 group A and B were subcutaneous injected with PBS (100 μl) and group C were subcutaneous injected with Der f crude protein (350μg) in PBS(100 μl) at 1-day intervals.One week after the last treatment ,group A,B and C were intranasally challenged with 50 μg Der f crude protein daily for seven days .Twenty-four hours after the last challenge , airway hyper-responsiveness ( AHR) was assessed by using whole-body plethysmography .Two days post challenged , mice were sacrificed and bronchoalveolar lavage fluid ( BALF) was collected.Number of the total cells and eosinophil was determined .Level of IL-4,IL-10 and IFN-γcytokines in the BALF and the su-pernatant of splenocyte culture was assayed by ELISA .Level of Der f specific IgE and histamine in the sera was determined by ELISA . Airway inflammation was analyzed by HE staining .Observation of the morphology and degranulation of mast cells was analyzed by tolui -dine blue staining.Results:Compared with group B,AHR and the lung inflammation in group C were greatly reduced (P<0.01). Numbers of total cells and eosinophils in BALF of group C were significantly lower than that of group B ( P<0.01 ) .Compared with group B, the observation of degranulation of mast cells was insignificant in group C .Compared with group B(IgE:1.905), the level of specific IgE was significantly lower in groups C (IgE:1.278)(P<0.01).The level of IL-4 in BALF of group C was significantly lower than that of group B(P<0.01).Compared with group A and B, the level of IL-10 in BALF was significantly higher in group C (P<0.01) and the level of IFN-γin BALF of group C was significantly higher than that of group A and B (P<0.01).Compared with group B, the level of IL-4 in cultured splenocytes was significantly lower in group C (P<0.01), and the level of IL-10 and IFN-γin cultured splenocytes of group C was significantly higher than that of group B (P<0.01).Compared with group B, the level of histamine in BALF was slightly lower in groups C (P<0.05), and the level of histamine in sera was significantly lower in groups C (P<0.05). Conclusion:The degranulation of mast cells of murine bronchial asthma model was suppressed after the desensitization therapy .

Objective To investigate the influence of anti-anxiety and depression treatment on quality life and plasma B-type natriuretic peptide (BNP) for chronic heart failure (CHF) patients.Methods Eighty-two patients with CHF were divided into control group and intervention group by random digits table method with 41 cases each.After admission,the patients were given conventional anti-heart failure treatment.The patients in intervention group received the standard therapy in combination with lorazepam,with depression plus fluoxetine,and psychological treatment.After treatment,the patients were evaluated by symptom checklist (SCL-90),determined the plasma BNP level and heart function indexes,and recorded the length of stay,number of hospitalization,and so on.Results After treatment the intervention group somatization,interpersonal barriers as well as depression,anxiety,hostility,terrorism,paranoia,psychotic and other emotions significantly alleviated,there were statistical differences compared with before treatment and after treatment of control group (P ＜ 0.05).There were no statistical differences in heart function indexes before treatment between the 2 groups (P ＞ 0.05).After treatment in control group,only the left ventricular ejection fraction was significantly higher than that before treatment,there was statistical difference (P ＜ 0.05).After treatment in intervention group compared with before treatment and after treatment in control group,the heart rate and left ventricular early diastolic and late peak velocity ratio were significantly lower [(71.0 ± 10.9) times/min vs.(82.5 ± 12.4) and (77.3 ± 10.1) times/min,1.26 ± 0.28 vs.1.38 ± 0.21 and 1.31 ± 0.33],left ventricular ejection fraction was significantly higher [(50.9 ± 6.9)％ vs.(39.2 ± 7.4)％ and (43.4 ± 7.8)％],there were statistical differences (P ＜ 0.05).The plasma BNP level was downtrend after treatment in the 2 groups,compared with the control group,after 5 d of treatment in intervention group was significant decrease,the changes was slowly after 14 d of treatment.There were statistical differences in plasma BNP level in 5,7,10 d of treatment between the 2 groups (P ＜ 0.05).The length of stay,number of hospitalization and fatality rate in intervention group were lower than those in control group [(6.0 ± 1.4)d/time vs.(10.0 ± 2.1) d/time,(1.6 ± 0.5) times/year vs.(3.2 ± 0.8) times/years,0 vs.9.8％ (4/41)],there were statistical differences (P ＜ 0.05).Conclusion Anti-anxiety and depression treatment can significantly improve cardiac function in CHF patients,shorter hospital stays,and improve quality of life.

Objective To investigate the distribution and antimicrobial resistance of pathogens from lower respira-tory tract in patients in intensive care unit (ICU),so as to provide reference for clinical treatment.Methods Distri-bution and antimicrobial susceptibility of pathogens isolated from ICU patients’sputum obtained through fiberbron-choscope between 2011 and 2014 were analyzed retrospectively.Results A total of 3 454 pathogenic strains were isolated between January 1 ,2011 and December 31 ,2014,the percentage of gram-negative bacteria,gram-positive bacteria,and fungi were 84.11 %,14.50%,and 1 .39% respectively.The detection rates of extended-spectrum β-lactamase-producing Escherichia coli and Klebsiella spp .in 2011 -2014 were 38.46% -73.33% and 26.95% -37.06% respectively. Enterobacteriaceae strains had low resistance rates to imipenem and meropenem (<20.00%);resistance of Acinetobacter baumannii was higher than Pseudomonas aeruginosa ,both had low resistance rates to amikacin(3.32%-37.16%);vancomycin-and linezolid-resistant strains were not found among Staphylo-coccus .In 2011 - 2014,detection rates of methicillin-resistant Staphylococcus aureus (MRSA)were 42.86% -61 .22%,methicillin-resistant coagulase-negative staphylococcus (MRCNS)were 86.96% - 91 .67%;resistance rates of Enterococcus faecium was higher than Enterococcus faecalis ,vancomycin-resistant strains were not found among Enterococcus faecalis and Enterococcus faecium ;the major fungus was Candida albicans .Conclusion Anti-microbial resistance of pathogens isolated from lower respiratory tract is getting more serious,clinicians should pay attention to non-antimicrobial infection control strategies in addition to rational use of antimicrobial agents.

Objective Lipid metabolism disorder can lead to male sterility, but its mechanism of affecting spermatogenesis and testis microenvironment remains unclear.This study aimed to investigate the effects of high fat diet on reproductive function and the expression of Ox-LDL within testis in male mice. Methods 16 C57BL/6 mice aged 8 weeks were divided into two groups by random number table method.High fat diet group was fed with high fat diet while normal fat diet group was fed with normal fat diet .At the end of 16 weeks, the levels of TG, TC, Ox-LDL and testosterone in serum were measured.The sperm concentration and motility from caudal epi-didymis were analyzed.The testis structure was observed by HE stai-ning.The localization and expression of Ox-LDL in testis were detec-ted by immunohistochemical technique. Results Compared with the normal diet group mice, the body weight (P<0.05) and the ser-um levels of TC [(2.31 ±0.33)mmol/L vs (6.54 ±0.31)mmol/L, P<0.01], Ox-LDL [(0.32 ±0.03)mg/L vs ( 0.44 ±0.06)mg/L, P<0.01] increased in high fat diet group mice significantly.The serum level of TG was of no significant difference(P>0.05).The serum level of testosterone[(3.64 ±0.43)mg/L vs (0.40 ±0.14) mg/L, P<0.01], the sperm concentration[(9.95 ±0.75)106/mL vs (5.66 ±1.51)106/mL, P<0.01] and the sperm motility[(54.69 ±17.84)%vs (32.48 ±5.80)%, P<0.01] decreased in high fat diet group significantly.HE staining also showed that the amount of Leydig cells, spermatids and spermatozoons reduced obvi-ously in high fat diet group.Immunohistochemistry staining showed that Ox-LDL was mainly distributed around Leydig cells of mice tes-tis.The expression of Ox-LDL in high fat diet group increased significantly ( P<0.01) . Conclusion The expression of Ox-LDL in Leydig cells of high fat diet C57BL/6J mice increases significantly, which may inhibit testosterone biosynthesis and affect spermatogen-ic function.

Objective To compare the curative effect and biomechanical performance of cannulated compression screw (CCS) and dynamic hip screw-blade (DHS-B) in the treatment of patients with femoral neck fracture.Methods Between February 2010 and February 2014,102 patients with femoral neck fracture were treated with CCS or DHS-B at our department.They were 54 males and 48 females,aged from 15 to 86 years.There were 30 subcapital fractures,51 transcervical ones and 21 base ones.CCS was used in 60 patients and DHS-B in 42.In-hospital data were collected retrospectively to compare the curative effects in 2 groups.Furthermore,femoral neck fracture models were established using 12 adult cadaveric femoral specimens.The 12 models were randomized into 2 equal groups (n =6).Group A was subjected to fixation by 3 CCSs and group B to fixation by DHS-B.The 2 groups were compared in terms of axial loading test,rotation test and destructive axial loading test.Results The operation time (59.4 ± 20.2 min),incision size (4.1 ±0.6 cm) and intraoperative blood loss (25.9 ±9.9 mL) in the CCS group were significantly less than those in the DHS-B group (88.6±22.9 min,12.1 ±1.2cmand 156.7±107.1 mL) (P ＜0.05).The Harris hip score for the DHS-B group (91.9±9.8) was significantly higher than that for the CCS group (87.2 ± 9.2) (P ＜ 0.05).There were no significant differences between the 2 groups in hospital stay,partial weight-bearing time,or postoperative complications (P ＞ 0.05).At 500 N vertical loading,the stress values at both medial and lateral sides of the femur in group A were significantly smaller than those in group B (P ＜ 0.05).There were no significant differences between groups A and B in the average sinking displacement of femoral head or the torque at a torsion angle of 6° (P ＞ 0.05).The maximum load in group A (2,135 ±120 N) was significantly smaller than that in group B (2,986 ± 98 N) (P ＜ 0.05).Conclusion In treatment of femoral neck fracture,DHS-B fixation is obviously superior to CCS fixation,because the former is in better agreement with the femoral biomechanical property,and performs better in anti-rotation and anti-compression,leading to better functional recovery of the affected hip.

Objective:To explore the effect of short-term modularized training mode with clear milestones for case competition of standardized residency training.Methods:Surgical residents, who attended the first case competition of Guangdong Province in 2020, were trained by short-term modularized training mode with clear milestones. This mode contained four modules: SP consultation training, training of disease characteristics and principles of diagnosis and treatment summary, medical documents and case analysis writing training, and comprehensive emergency case training.Results:After the training, they all made great progress in consultation, interpretation of exam results, and first writing of course record, but less in medical documents and case analysis. The satisfaction rate was 100%, and they all considered informed consent writing for operation and formulating principles of diagnosis and treatment summary as difficult tasks.Conclusion:Short-term modularized training mode with clear milestones might improve the level of consultation, interpretation of exam results, and first writing of course record effectively. While, the difficult point is clinical thinking about the surgical treatment, which must be trained more in routine work.

Objective To study the clinical efficacy of caffeine in premature infants receiving mechanical ventilation and related complications .Method From January 2014 to September 2016, preterm infants (28w≤GA<33w) treated with nasal continuous positive airway pressure (NCPAP) or conventional mechanical ventilation ( CMV ) in neonatal intensive care unit were studied .They were randomly assigned into the caffeine group and the control group .The caffeine group received caffeine when NCPAP ventilation was applied or adjusting to synchronized intermittent mandatory ventilation ( SIMV) mode.The control group was injected with 5%glucose without caffeine .The t test andχ2 test were used to analyze the clinical efficacy and related complications .Result A total of 96 patients receiving NCPAP ventilation were collected ( birth weight between 1300~2100 g), including 51 cases in caffeine group and 45 cases in the control group. 84 cases received CMV ventilation (birth weight between 1000~1499 g), with 43 cases in the caffeine group and 41 cases the control group.Among the NCPAP infants, the incidence of failure to withdraw ventilator (0% vs.13.3%) and the incidence of bronchopulmonary dysplasia (3.9% vs.17.8%) were lower in the caffeine group than the control group .The duration of assisted ventilation and hospital stay in the caffeine group were shorter than the control group [(6.2 ±3.1) d vs.(8.2 ±3.2) d, (16.3 ±8.7) d vs. (19.5 ±9.2) d], the differences were statistically significant (P<0.05).Among the CMV infants, the incidence of failure of A/C to SIMV mode transition and bronchopulmonary dysplasia were lower in the caffeine group than the control group and the duration of assisted ventilation and hospital stay were shorter . The differences were statistically significant ( P <0.05 ) . No differences were found in the related complications in each group ( P>0.05) . Conclusion Caffeine can help reduce the incidences of withdrawal failure, bronchopulmonary dysplasia , ventilation duration and hospital stay when using NCPAP and CMV ventilation.

Objective:The consensus molecular subtype (CMS) classification is based on the gene expression profiles, This article attempts to conduct a preliminary exploration and analysis of the clinical features of different CMS patients. We can give an individualized diagnosis and treatment for the patients.Methods:Seven GSE series of colorectal cancer gene expression profiles were downloaded by R software from the GEO database. A total of 1414 patients were included. Using the specific computational method published by Peter in 2017, the patients were divided into four groups: CMS1, CMS2, CMS3 and CMS4. The measurement data is expressed by Mean± SD, and the count data is expressed by n(%). The software of SPSS 18.0 was used to conduct analysis. Results:CMS1 tumors originated in the right colon (77.4%), while CMS2 mostly originated in the left colon (72.8%). The proportion of T 4 stage in CMS2 was 16.6%, while in the other three types was 23.3%, 29.3% and 24% respectively; the proportion of distant metastasis of CMS1 was the lowest (3.5%), while the distant metastasis rate of CMS4 was 18.2%. The KRAS mutation rates in CMS1 and CMS2 were 25.6% and 30.3% respectively, while in CMS3 it was up to 73.9%; the BRAF mutation rate in CMS1 was 45.5%, while the other three mutation rates were 0.6%, 6.2% and 5.9%, respectively; The average mutation rate of APC was 59.45%. Overall survival and progression-free survival analysis showed that the CMS4 interstitial type was the worst, while the CMS2 classic type had the best relative prognosis. Conclusions:CMS1 immunotype, the tumor has origin from right colon in female patients. MSI-H is often accompanied by BRAF gene mutation, this type patients has a refractory treatment response and poor prognosis. The classic CMS2 type is characterized by APC deletion mutation and Wnt activation, with good therapeutic effect and good prognosis. CMS3 metabolic type often harbor KRAS mutation, anti-EGFR treatment is not sensitive. Although this type is prone to relapse, but the chemotherapy is effective, so the overall survival prognosis is acceptable. CMS4 interstitial type with left colon tumor is prevalence. due to the activation of TGF-β and enhanced angiogenesis, this type tumor is prone to metastasis to distant site and has the worst prognosis.

Background: Gastroenteropancreatic neuroendocrine neoplasm (GEP-NEN) is a rare heterogeneous tumor. Liver metastasis seriously affects the prognosis of GEP-NEN. However, few tools are existed to predict GEP-NEN complicated with synchronous liver metastasis. Aims: To analyze the risk factors of synchronous liver metastasis in patients with GEP-NEN and establish a nomogram to predict synchronous liver metastasis in patients with GEP-NEN. Methods: A total of 10 973 pathologically confirmed patients with GEP-NEN from Jan. 2010 to Dec. 2017 were collected from SEER database and divided randomly into training set (n=7 511) and test set (n=3 462). Both groups were divided into liver metastasis group and non-liver metastasis group according to the occurrence of liver metastasis. Multifactorical logistic regression analysis was used to identify the risk factors of liver metastasis in patients with GEP-NEN. R software was used to establish and verify the nomogram of liver metastasis in GEP-NEN patients. Results: Liver metastasis was associated with gender, age, race, primary tumor site, degree of differentiation, tumor diameter, T3/4 stage, and lymph node metastasis in patients with GEP-NEN. The results of multivariate logistic regression analysis showed that primary tumor site (small intestine and pancreas), differentiation degree (poorly differentiated and undifferentiated), diameter of tumor ≥ 5 cm, T3/4 stage and lymph node metastasis were independent risk factors affecting liver metastasis in patients with GEP-NEN (P< 0.001). The concordance index of internal validation for nomogram was 0.838 (95% CI: 0.826-0.849), and the concordance index of external validation was 0.847 (95% CI: 0.829-0.864). Conclusions: GEP-NEN patients with primary tumor site in small intestine or pancreas, poor differentiation and undifferentiation, diameter of tumor ≥5 cm, T3/4 stage and lymph node metastasis are more likely to develop liver metastasis which suggested that such patients need to be alert for the occurrence of liver metastasis and need more aggressive treatment. The calibration curves fits are good for both the training and test sets, and can help clinicians to make individualized prediction for whether the GEP-NEN patient has synchronous liver metastasis at the initial diagnosis.