Chloroplast genomic characterization and phylogenetic analysis of Castanopsis hystrix / 生物工程学报

The structure and size of the chloroplast genome of Castanopsis hystrix was determined by Illumina HiSeq 2500 sequencing platform to understand the difference between C. hystrix and the chloroplast genome of the same genus, and the evolutionary position of C. hystrix in the genus, so as to facilitate species identification, genetic diversity analysis and resource conservation of the genus. Bioinformatics analysis was used to perform sequence assembly, annotation and characteristic analysis. R, Python, MISA, CodonW and MEGA 6 bioinformatics software were used to analyze the genome structure and number, codon bias, sequence repeats, simple sequence repeat (SSR) loci and phylogeny. The genome size of C. hystrix chloroplast was 153 754 bp, showing tetrad structure. A total of 130 genes were identified, including 85 coding genes, 37 tRNA genes and 8 rRNA genes. According to codon bias analysis, the average number of effective codons was 55.5, indicating that the codons were highly random and low in bias. Forty-five repeats and 111 SSR loci were detected by SSR and long repeat fragment analysis. Compared with the related species, chloroplast genome sequences were highly conserved, especially the protein coding sequences. Phylogenetic analysis showed that C. hystrix is closely related to the Hainanese cone. In summary, we obtained the basic information and phylogenetic position of the chloroplast genome of red cone, which will provide a preliminary basis for species identification, genetic diversity of natural populations and functional genomics research of C. hystrix.

CRHR1 mediates the transcriptional expression of pituitary hormones and their receptors under hypoxia.

Hypothalamus-pituitary-adrenal (HPA) axis plays critical roles in stress responses under challenging conditions such as hypoxia, via regulating gene expression and integrating activities of hypothalamus-pituitary-targets cells. However, the transcriptional regulatory mechanisms and signaling pathways of hypoxic stress in the pituitary remain to be defined. Here, we report that hypoxia induced dynamic changes in the transcription factors, hormones, and their receptors in the adult rat pituitary. Hypoxia-inducible factors (HIFs), oxidative phosphorylation, and cAMP signaling pathways were all differentially enriched in genes induced by hypoxic stress. In the pituitary gene network, hypoxia activated c-Fos and HIFs with specific pituitary transcription factors (Prop1), targeting the promoters of hormones and their receptors. HIF and its related signaling pathways can be a promising biomarker during acute or constant hypoxia. Hypoxia stimulated the transcription of marker genes for microglia, chemokines, and cytokine receptors of the inflammatory response. Corticotropin-releasing hormone receptor 1 (CRHR1) mediated the transcription of Pomc, Sstr2, and Hif2a, and regulated the function of HPA axis. Together with HIF, c-Fos initiated and modulated dynamic changes in the transcription of hormones and their receptors. The receptors were also implicated in the regulation of functions of target cells in the pituitary network under hypoxic stress. CRHR1 played an integrative role in the hypothalamus-pituitary-target axes. This study provides new evidence for CRHR1 involved changes of hormones, receptors, signaling molecules and pathways in the pituitary induced by hypoxia.

Mathematical model of postharvest variation in tomato color based on optimized response surface methodology.

BACKGROUND: Manual inspection and instrumentation form the traditional approach to determining tomato color but these methods only determine tomato color at a given moment and cannot predict dynamically how tomato color varies during storage and transportation. Such methods thus cannot help suppliers and retailers establish good management practices for the flexible control of tomato maturity, accurate judgment of market positioning in the industry, or during distribution and marketing. To address this shortcoming, this work first investigates how tomato color parameters (a* and h°) evolve through the various stages of maturity (green, turn, and light red) under different storage conditions. Based on experimental results, it develops an optimized response-surface model (RSM) by using differential evolution to predict how tomato color varies during storage. RESULTS: Tomatoes are more likely to change color at high temperatures and under conditions of high humidity. Temperature affects tomato color more strongly than humidity. The accuracy of the RSM was confirmed by a good agreement with experiments. All determination coefficients R2 of the RSMs for a* and h° are greater than 0.91. The mean absolute errors for a* and h° are 3.8112 and 5.6500, respectively. The root mean square errors for a* and h° are 4.6840 and 6.9198, respectively. CONCLUSION: This research reveals how storage temperature and humidity affect the postharvest variations in tomato color and thus establishes a dynamic model for predicting tomato color. The proposed RSM provides a reliable theoretical foundation for dynamic, nondestructive monitoring of tomato ripeness in the cold chain. © 2021 Society of Chemical Industry.

The effect of combining transcranial magnetic stimulation with escitalopram on neurological functioning, depression and serum inflammatory factors after a stroke / 中华物理医学与康复杂志

Objective:To investigate the effect of combining repeated high-frequency transcranial magnetic stimulation (rTMS) with escitalopram in treating the neurological functioning and post-stroke depression of stroke survivors.Methods:Eighty persons with post-stroke depression were randomly divided into an observation group and a control group, each of 40. The control group was treated with oral escitalopram, while the observation group also received transcranial magnetic stimulation at 5Hz. The magnetic stimulation intensity was 80% of each person′s resting motion threshold. The rTMS was administered once a day, 5 days a week for 8 weeks. Neurological functioning and depression were evaluated using National Institutes of Health stroke scoring (NIHSS) and the Hamilton depression scale before and after the 8 weeks of treatment. The levels of tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2) and interleukin-6 (IL-6) in serum samples from the two groups were detected using enzyme-linked immunosorbent assays.Results:After treatment, the NIHSS and Hamilton scores and TNF-α, IL-2 and IL-6 levels in both groups had improved significantly, but in each case they were significantly better in the treated group, on average.Conclusion:Supplementing escitalopram with high-frequency transcranial magnetic stimulation more effectively improves depression and neurological functioning after a stroke.

Functional Diversity of p53 in Human and Wild Animals.

The common understanding of p53 function is a genome guardian, which is activated by diverse stresses stimuli and mediates DNA repair, apoptosis, and cell cycle arrest. Increasing evidence has demonstrated p53 new cellular functions involved in abundant endocrine and metabolic response for maintaining homeostasis. However, TP53 is frequently mutant in human cancers, and the mutant p53 (Mut-p53) turns to an "evil" cancer-assistant. Mut-p53-induced epithelial-mesenchymal transition (EMT) plays a crucial role in the invasion and metastasis of endocrine carcinomas, and Mut-p53 is involved in cancer immune evasion by upregulating PD-L1 expression. Therefore, Mut-p53 is a valuable treatment target for malignant tumors. Targeting Mut-p53 in correcting sequence and conformation are increasingly concerned. Interestingly, in wild animals, p53 variations contribute to cancer resistant and high longevity. This review has discussed the multiple functions of p53 in health, diseases, and nature evolution, summarized the frequently mutant sites of p53, and the mechanisms of Mut-p53-mediated metastasis and immune evasion in endocrine cancers. We have provided a new insight for multiple roles of p53 in human and wild animals.

Post-marketing multi-center safety surveillance of inactivated enterovirus A71 vaccine (Vero cell) / 中华预防医学杂志

Objective@#To evaluate the post-marketing safety profiles of the inactivated enterovirus type 71 (EV-A71) vaccine (Vero cell) after routine inoculation.@*Methods@#Eleven cities of Zhejiang Province, Fengtai district of Beijing, Qinnan district, two counties as Pingle and Pingguo of Guangxi Zhuang Autonomous Region, and Dongtai city of Jiangsu Province were selected as the field sites. A total of 45 239 subjects were enrolled in this study from children who seeked the vaccination of EV-A71 vaccine during the period from July, 2016 to June, 2018. Different sampling method were adopted in different sites. All vaccinated children were invited to participate in the study in Fengtai and Dongtai, however, systematic sampling method were adopted in other sites. Active surveillance was conducted and information about adverse reactions (ARs) occurred in 30 min, 3 d and 30 d following each dose of EV-A71 immunization was collected by field observation, phone-call or face-to-face interview. The incidence of ARs in different types, symptoms and grades were described.@*Results@#In total, there were 45 239 children who received 71 243 doses EV-A71 vaccine. The overall incidence of ARs was 1.079% (769 doses), with the highest incidence of 1.182% (177/14 973) in 5-11 month group and the lowest incidence of 0.849% (18/2 119) in ≥ 36 month group among different age groups. There was a higher incidence in solicited ARs, which was 1.047% (746 doses). The incidences of grade 1 and grade 2 ARs were also higher, which were 0.404% (288 doses) and 0.554% (395 doses), respectively. No grade 4 ARs occurred. The doses of the first and the second vaccination was 40 736 and 30 507, respectively, and the incidences of ARs were 1.281% (522 doses) and 0.810% (247 doses). Also, the incidences of ARs were 0.091% (37 doses) and 0.043% (13 doses) in local, and 1.168% (476 doses) and 0.760% (232 doses) in system. The symptoms of ARs after the two doses of vaccination were basically the same. Redness at the injection site was the most common local ARs after each dose vaccination, with doses of 24 and 11, while fever was the most common systemic ARs, with doses of 362 and 190. Moreover, ARs mainly occurred in 30 min to 3 d after each dose vaccination, with incidence of 1.016% (414 doses) and 0.698% (213 doses) in the first and second dose, respectively.@*Conclusion@#The ARs had a low incidence after vaccination in children and most were mild or moderate. EV-A71 vaccine with good safety is suitable for inoculation in a large scale.

Artesunate attenuate chronic graft-versus-host disease by regulating Th17/Treg balance / 中华血液学杂志

Objective: To investigate the effects of artesunate treatment on chronic graft-versus-host disease (cGVHD). Methods: Recipient BALB/c mice received 8 × 10(6) bone marrow cells with 8×10(6) spleen cells from B10D2 mice. Artesunate solubilized in acetone was injected intraperitoneally every day at the dose of 1 mg/kg at Day 28 after BMT. The clinical scores, survival and histopathological damage were analyzed. The frequency of Th17 and Tregs in PB and spleens from the mice were evaluated by flow cytometry. In addition, CD4(+) T cells from the spleens of mice were cultured in vitro, then stimulated with artesunate, the frequency of Th17 and Tregs in these splenocytes were evaluated by flow cytometry. Results: Artesunate administration diminished clinical and histopathological damage, and improved the survival of cGVHD mice[(46.57±7.83)% vs (55.71±6.99)%, χ(2)=5.457, P=0.020]; Artesunate contributed to Tregs development [(4.45±0.04)% vs (8.40±0.23)%, t=15.679, P<0.001; (6.62±0.24)% vs (10.48±0.48)%, t=6.587, P=0.003] while decreased Th17 cells [(1.51±0.18)% vs (0.58±0.19)%, t=7.233, P<0.001; (1.48±0.38)% vs (0.71±0.18)%, t=3.653, P=0.011] expressions in both PB and spleens, and decreased the Th17/Treg ratio (0.34±0.05 vs 0.09±0.03, t=7.621, P=0.002; 0.19±0.03 vs 0.06±0.02, t=6.993, P=0.002). Moreover, artesunate suppressed the Th17 cells expressions [(0.82±0.37) % vs (3.39±1.22) %, t=4.044, P=0.007] and contributed to Tregs development [(34.63±1.29) % vs (14.28±1.69) %, t=19.119, P<0.001], and also decreased the Th17/Treg ratio (0.24±0.09 vs 0.02±0.01, t=4.780, P=0.003) in vitro. Conclusions: Artesunate suppressed the Th17 cells expressions and contributed to Tregs development, which provided new sights into the development of a novel drug for cGVHD, e.g., artemisinin.

Treatment of proximal humeral fractures with proximal humeral locking system plate via an intertubercular sulcus approach / 中国骨伤

OBJECTIVE@#To observe the effect of proximal humerus internal locking system (PHILOS) plate in the treatment of Neer 2 and 3 part fractures of proximal humerus through tuberosity approach.@*METHODS@#From July 2015 to January 2018, 15 cases of proximal humerus Neer 2 and 3 fractures were treated with PHILOS plate through intertubercular sulcus approach. There were 7 males and 8 females. The age ranged from 23 to 67 years old, with an average of 46 years old. There were 5 cases on the left side and 10 cases on the right side (including 7 cases of Neer 2 partial fracture and 8 cases of Neer 3 partial fracture). X-ray films and CT were taken before and after operation to assess fracture location and fracture healing. Clinical evaluation included Constant-Murley shoulder function score and operative complications. All 15 patients were treated with the PHILOS. Constant-Murley score was used to evaluate shoulder function after operation.@*RESULTS@#All the 15 cases were followed up, and the duration ranged from 14 to 36 months. All the fractures healed and the healing time ranged from 14 to 26 weeks, averaged 19.1 weeks. There were no complications such as humeral head necrosis, axillary nerve injury and fracture nonunion after operation. At the 3rd month after operation, the shoulder function score of Constant-Murley ranged from 72 to 94 points, with an average of 81 points; 2 cases got an excellent result and 13 good.@*CONCLUSIONS@#The PHILOS plate for the treatment of proximal humerus fractures has the advantages of simple operation, small injury and quick recovery of shoulder joint function.

Clinical significance of coagulase-negative Staphylococci isolated from blood cultures / 中华传染病杂志

Objective To analyze clinical characteristics of bloodstream infections caused by coagulase-negative Staphylococci (CNS) and antibiotic resistance of the bacteria,so that to provide basis for the clinical diagnosis and treatment.Methods A retrospective analysis of CNS in blood cultures collected from 108 hospitalized patients in Puai Hospital of Tongji Medical College from January 2016 to December 2017 was performed.The antimicrobial susceptibilities were tested by Kirby-Bauer method and E test method.For measurement variables,normally distributed variables were compared using t test,and non-normal distributed data were compared using Mann-Whitney U test.Categorical variables were compared using x2 test.Results Of the 108 patients,66 were male and 42 were female;the age range was 26 to 98 years and the average was 49 years.According to the criteria for bacteremia,36 of 108 (33.3％) patients with CNS-positive blood cultures were diagnosed with bacteremia and 72 (66.7％) cases were contaminated.CNS bacteremia mainly occurred in the intensive care unit and nephropathy ward.Among them,23 (62.2％) patients were catheter-related blood stream infections,and 11 (29.7 ％) patients were dialysis catheter-related bloodstream infections.Fifteen of 36 (41.7％) strains were isolated within 48 hours of admission.The level of serum procalcitonin (PCT) for bacteremia patients was 2.56 (1.44,7.60) μg/L,and that was 0.13 (0.05,0.23) μg/L in contaminated patients.The difference was statistically significant (Z=8.097,P＜0.05).The white blood cell count of patients with bacteremia was (11.50±4.54) × 109/L,and that was (10.61 ±5.00) × 109/L for contaminated patients.There was no statistical significance (t=0.895,P＞0.05).After antibiotic treatment,26 of 36 bacteremia patients were survived.The PCT levels before antibiotic treatment were 2.05 (1.42,4.32) μg/L,and 0.24 (0.07,0.61) μg/L after antibiotic treatment.Serum PCT was decreased significantly after antibiotic treatment (Z=4.457,P＜0.05).The PCT levels of 10 deaths within 28 days before antibiotic treatment were 4.78 (1.51,19.75) μg/L,whereas 22 (6.40,55.75) μg/L,after antibiotic treatment.The PCT was increased significantly after antibiotic treatment (Z=2.497,P＜0.05).No significant difference was found in PCT between survivors and deaths within 28 days (Z=0.300,P＞0.05).No significant difference was found in white blood cell count between survivors and deaths at 28 days (t=0.771,P＞0.05).There was no statistical difference of the anti-bacterial drug susceptibility between pathogens and contaminants (P＞0.05).All strains were sensitive to vancomycin,teicoplanin and linezolid.Conclusions The incidence of CNS contamination in blood culture is relatively high.It is important to distinguish true bacteraemia from contamination by a review of the clinical and laboratory indicators.PCT is of clinical value to indicate CNS infection and to monitor therapeutic effect.

Analysis of pathogen spectrum and antimicrobial resistance of clinical common organisms from twelve teaching hospitals in 2016 / 中华检验医学杂志

Objective To investigate the spectrum and antimicrobial resistance of major pathogens causing nosocomial infections in China, 2016. Methods Non-duplicated nosocomial cases as well as pathogens causing bloodstream infections ( BSI) , hospital-acquired pneumonia ( HAP) and intra-abdominal infections ( IAI ) from 12 teaching hospitals across China were collected. The minimum inhibitory concentrations (MICs) of important clinical common strains were determined by agar dilution method or broth microdilution method. The CLSI M100-S27 criteria was used for interpretation. Data were analyzed by using WHONET-5. 6 software. Results A total of 2060 cases were collected, including 894 cases from BSI, 630 cases from HAP and 536 cases from IAI. The MICs of 1896 important clinical common strains were determined. Escherichia coli and Klebsiella pneumoniae were the most prevalent pathogens causing BSI and IAI, while Acinetobacter baumanii and Pseudomonas aeruginosa were dominated in HAP. All Staphylococcus aureus were susceptible to tigecycline, linezolid, daptomycin and glycopeptides. Methicillin-resistant S. aureus accounted for 44. 4％ ( 75/169 ) of all the S. aureus. The rate of methicillin-resistant coagulase-negative staphylococci was 80. 9％ ( 72/89 ) . No Enterococcus strains were found resistant to tigecycline, linezolid or daptomycin. Vacomycin resistant enterococcus was found in Enterococcus faecium, accounting for 1. 8％ ( 2/111 ) of all E. faecium strains. Tigecycline, meropenem, amikacin, imipenem, and polymyxin B exhibited high potency against Enterobacteriaceae and the susceptibility rates were 96. 6％(865/895), 94. 3％ (859/911), 94. 2％ (858/911), 94. 1％ (857/911), and 91. 6％ (820/895), respectively. The prevalence of extended-spectrum β-lactamase was 58. 4％ ( 263/450 ) in E. coli and 28. 6％ ( 84/294 ) in K. pneumonia. The rate of carbapenem resistant K. pneumonia and E. coli was 15. 3％ ( 45/294 ) and 1. 8％ ( 8/450 ) , respectively. The percentage of polymyxin B resistant K. pneumonia and E. coli was 4. 1％ ( 12/294 ) and 4. 4％ ( 20/450 ) , respectively. The rate of tigecycline resistant K. pneumonia and E. coli was 2. 4％ ( 7/294 ) and 0. 2％ ( 1/450 ) , respectively. A. baumanii showed low susceptibility to the antimicrobial agents except tigecycline ( 91. 4％, 235/257 ) and polymyxin B (100％, 257/257). The rate of carbapenem resistant A. baumanii was 80. 5％ (207/257). The rate of carbapenem resistant P. aeruginosa was 31. 7％ ( 59/186 ) . Polymyxin B and amikacin demonstrated high antibacterial activity against P. aeruginosa with susceptility rate of 100％ ( 186/186 ) and 90. 9％ ( 169/186), respectively. Conclusions Nosocomial pathogens showed high susceptibilities against tigecycline and polymyxin B. Antimicrobial resistance in A. baumannii is a serious problem. The prevalence of carbapenem-resistant Enterobacteriaceae and polymyxin B resistant Enterobacteriaceae has increased, which should be monitored continuously in China.

Microbiological profiles of pathogens causing nosocomial bacteremia in 2011, 2013 and 2016 / 生物工程学报

To dynamically investigate the distribution and antimicrobial resistance profiles of bacteremia pathogens isolated from different regions in China in 2011, 2013 and 2016. Non-repetitive isolates from nosocomial bloodstream infections were retrospectively collected and detected for antimicrobial susceptibility tests (AST) by agar dilution or microbroth dilution methods. Whonet 5.6 was used to analyze the AST data. Among 2 248 isolates, 1 657 (73.7%) were Gram-negative bacilli and 591 (26.3%) were Gram-positive cocci. The top five bacteremia pathogens were as follows, Escherichia coli (32.6%, 733/2 248), Klebsiella pneumoniae (14.5%, 327/2 248), Staphylococcus aureus (10.0%, 225/2 248), Acinetobacter baumannii (8.7%, 196/2 248) and Pseudomonas aeruginosa (6.2%, 140/2 248). Colistin (96.5%, 1 525/1 581, excluding innate resistant organisms), tigecycline (95.6%, 1 375/1 438, excluding innate resistant organisms), ceftazidine/clavulanate acid (89.2%, 1 112 /1 246), amikacin (86.4%, 1 382/1 599) and meropenem (85.7%, 1 376/1 605) showed relatively high susceptibility against Gram-negative bacilli. While tigecycline, teicoplanin and daptomycin (the susceptibility rates were 100.0%), vancomycin and linezolid (the susceptibility rates were 99.7%) demonstrated high susceptibility against Gram-positive cocci. The prevalence of extended-spectrum β-lactamases (ESBLs)-producing Enterobacteriaceae were 50.6% (206/407), 49.8% (136/273) and 38.9% (167/429) in 2011, 2013 and 2016 respectively; carbapenem-non-susceptible Enterobacteriaceae were 2.2% (9/408), 4.0% (16/402) and 3.9% (17/439) in 2011, 2013 and 2016 respectively; The prevalence of multidrug-resistant A. baumannii (MDRA) was 76.4% (55/72) in 2011, 82.7% (43/52) in 2013 and 87.5% (63/72) in 2016, respectively. The prevalence of multidrug-resistant P. aeruginosa (MDRP) was 9.8% (5/51) in 2011, 20.0% (7/35) in 2013 and 13.0% (7/54) in 2016, respectively. The prevalence of methicillin-resistant S. aureus (MRSA) was 51.9% (41/79) in 2011, 29.7% (19/64) in 2013 and 31.7% (26/82) in 2016, respectively. The prevalence of high level gentamicin resistance (HLGR) of Enterococcus faecium and Enterococcus faecalis were 43.2% (48/111) and 40.9% (27/66), respectively. The predominant organism of carbapenem-non-susceptible Enterobacteriaceae was K. pneumoniae with its proportion of 57.1% (24/42). Among 30 tigecycline-non-susceptible Enterobacteriaceae, K. pneumoniae was the most popular organism with 76.7% (23/30). Among 39 colistin-resistant Enterobacteriaceae, E. coli, Enterobacter cloacae and K. pneumoniae were constituted with the percent of 43.6 (17/39), 35.9 (14/39) and 15.4 (6/39), respectively. The Gram-negative bacilli (E. coli and K. pneumoniae were the major organisms) were the major pathogens of nosocomial bacteremia, to which tigecycline, colistin and carbapenems kept with highly in vitro susceptibility. Whereas, among the Gram-positive cocci, S. aureus was the top 1 isolated organism, followed by E. faecium, to which tigecycline, daptomycin, linezolid, vancomycin and teicoplanin kept with highly in vitro susceptibility. Isolation of colistin-resistant Enterobacteriaceae, tigecycline-non-susceptible Enterobacteriaceae, linezolid- or vancomycin-non-susceptible Gram-positive cocci suggests more attention should be paid to these resistant organisms and dynamic surveillance was essential.

Meta plate and cannulated screw fixation for treatment of type Letenneur III lateral Hoffa fracture through posterolateral approach / 中国骨伤

<p><b>OBJECTIVE</b>To explore clinical effect of meta plate with cannulated screw fixation in treating Letenneur III lateral Hoffa fracture through posterolateral approach.</p><p><b>METHODS</b>From June 2012 to June 2016, 12 patients with Letenneur III lateral Hoffa fracture were treated with meta plate and cannulated screw fixation through posterolateral approach, including 8 males and 4 females;aged from 22 to 66 years with an average of 42 years old;and 5 cases on left side, 7 cases on right side. Preoperative and postoperative X-ray and CT were taken before and after operation to assess location of fracture and fracture healing. Letenneur scoring standard was used to evaluate clinical effects. All patients were treated with meta plate combined with cannulated screws.</p><p><b>RESULTS</b>All incisions healed at stage I. The fractures were healed from 11 to 32 weeks with an average of 17.2 weeks. No internal fixation loosening, displacement and necrosis of femoral condyle and deep vein thrombosis of lower extremity occurred. All patients were followed up from 8 to 24 months with an average of 12.8 months. At the latest follow-up, 7 cases got excellent results, 4 good and 1 moderate according to Letenneur scoring standard.</p><p><b>CONCLUSIONS</b>Posterlateral approach is easy to expose, meta plate fixation with cannulated screws has advantages of strong internal fixation, high fracture healing rate, early knee joint function exercise and good recovery of joint function.</p>

Identification of the paradoxical effect of caspofungin and micafungin on Candida albicans growth in vitro using a growth curve methodology.

BACKGROUND: The paradoxical effect of fungi, (which is a special phenomenon that certain Candida spp. might exhibit augmented growth in the presence of echinocandin at some concentration higher than the minimum inhibitory concentration) would be better illustrated through growth curves. MATERIALS AND METHODS: We investigated the paradoxical effect of caspofungin and micafungin on the growth and cell viability of 38 clinical isolates of Candida albicans using a microbial sensitivity test and growth curve methodology. RESULTS: When cells were incubated in the presence of antifungal agents for 48 h, the paradoxical effect was observed in 50% and 47.4% of the isolates incubated with caspofungin and micafungin, respectively. Growth curves indicated the occurrence of the paradoxical effect varied between different echinocandins. Moreover, increased turbidity in the sensitivity test did not correlate with increased fungal cell growth. For caspofungin, the paradoxical effect appeared earlier than in the microbial sensitivity test, while for micafungin, the effect observed in the microbial sensitivity test was not a true paradoxical effect. CONCLUSION: The growth curve methodology is more accurate for evaluating the occurrence of the paradoxical effect in comparison with the microbial sensitivity test.

The integration of multiple signaling pathways provides for bidirectional control of CRHR1 gene transcription in rat pituitary cell during hypoxia.

Hypoxia upregulates hypothalamic corticotrophin releasing hormone (CRH) and its receptor type-1 (CRHR1) expression and activates the HPA axis and induces hypoxic sickness and behavioral change. The transcriptional mechanism by which hypoxia differently regulates CRHR1 expression remains unclear. Here we report hypoxia time-dependently induced biphasic expression of CRHR1mRNA in rat pituitary during different physiological status. Short exposure of gestational dams to hypoxia reduced CRHR1mRNA in the pituitary of P1-P14 male rat offspring. A short- and prolonged-hypoxia evoked biphasic response of CRHR1mRNA characterized initially by decreases and subsequently by persistent increases, mediated by a rapid negative feedback via CRHR1 signaling and positive transcriptional control via NF-κB, respectively. Further analysis of CRHR1 promoter in cultured primary anterior pituitary and AtT20 cells showed that c-Jun/AP-1 delivered negative while HIF-1α and NF-κB delivered positive control of transcription at CRHR1 promoter. The negative and positive inputs are integrated by hypoxic initiation and duration in CRHR1 transcription.

Utilization of a Wheat660K SNP array-derived high-density genetic map for high-resolution mapping of a major QTL for kernel number.

In crop plants, a high-density genetic linkage map is essential for both genetic and genomic researches. The complexity and the large size of wheat genome have hampered the acquisition of a high-resolution genetic map. In this study, we report a high-density genetic map based on an individual mapping population using the Affymetrix Wheat660K single-nucleotide polymorphism (SNP) array as a probe in hexaploid wheat. The resultant genetic map consisted of 119 566 loci spanning 4424.4 cM, and 119 001 of those loci were SNP markers. This genetic map showed good collinearity with the 90 K and 820 K consensus genetic maps and was also in accordance with the recently released wheat whole genome assembly. The high-density wheat genetic map will provide a major resource for future genetic and genomic research in wheat. Moreover, a comparative genomics analysis among gramineous plant genomes was conducted based on the high-density wheat genetic map, providing an overview of the structural relationships among theses gramineous plant genomes. A major stable quantitative trait locus (QTL) for kernel number per spike was characterized, providing a solid foundation for the future high-resolution mapping and map-based cloning of the targeted QTL.

Overexpression of miRNA-199a-3p targets at expression of MAP3K4 in gastric cancer / 实用医学杂志

Objective To investigate the effect of miRNA-199a-3p overexpression on the expression of MAP3K4 protein in gastric cancer. Methods 35 gastric cancers and the matched adjacent tissue specimens were collected. Expression of miRNA-199a-3p and MAP3K4 were detected by stem-loop real-time reverse transcription polymerase chain reaction and western blot. Cell transfection was employed to explore the regulation of miRNA-199a-3p on MAP3K4 gene. Luciferase reporter assay was performed to identify target MAP3K4 gene. Results Com-pared with the adjacent tissue specimens ,miRNA-199a-3p was upregulated in the gastric cancers ,and MAP3K4 protein was down-regulated in the gastric cancers. Cells transfected with miR-199a-3p mimics showed lower MAP3K4 protein. MAP3K4 was identified as target gene of miR-199a-3p. Conclusions miRNA-199a-3p acts as an oncogene in gastric cancer and functions by targeting MAP3K4.

Overexpression of miRNA-199a-3p targets at expression of MAP3K4 in gastric cancer / 实用医学杂志

Objective To investigate the effect of miRNA-199a-3p overexpression on the expression of MAP3K4 protein in gastric cancer. Methods 35 gastric cancers and the matched adjacent tissue specimens were collected. Expression of miRNA-199a-3p and MAP3K4 were detected by stem-loop real-time reverse transcription polymerase chain reaction and western blot. Cell transfection was employed to explore the regulation of miRNA-199a-3p on MAP3K4 gene. Luciferase reporter assay was performed to identify target MAP3K4 gene. Results Com-pared with the adjacent tissue specimens ,miRNA-199a-3p was upregulated in the gastric cancers ,and MAP3K4 protein was down-regulated in the gastric cancers. Cells transfected with miR-199a-3p mimics showed lower MAP3K4 protein. MAP3K4 was identified as target gene of miR-199a-3p. Conclusions miRNA-199a-3p acts as an oncogene in gastric cancer and functions by targeting MAP3K4.

Mini-locking plates for the treatment of Regan-Morrey type III fracture of ulnar coronoid process through anterior approach of elbow joint / 中国骨伤

<p><b>OBJECTIVE</b>To explore clinical effects of mini-locking plates for the treatment of Regan-Morrey type III fractures of ulnar coronoid process through an anterior approach of elbow joint.</p><p><b>METHODS</b>A retrospective analysis on 12 patients with Regan-Morrey type III fractures of the ulnar coronoid process was performed from January 2011 to June 2014, who were treated with unini-locking plates through the anterior approach of elbow joint. There were 7 males and 5 females, ranging in age from 23 to 65 years old, averaged 43 years old. Four patients had fractures on the left and 8 patients had fractures on the right. The X-ray films were taken to evaluate the location and healing of the fracture before and after operation. Clinical evaluation included analysis on surgical complications, range of motion and Mayo elbow function score. All the patients were treated with mini-locking plates, and the elbow joint was stable after operation.</p><p><b>RESULTS</b>All the patients were followed up, and the duration ranged from 14 to 36 months. All the fractures were healed, and the healing time ranged from 12 to 20 weeks. The average healing time was 15.6 weeks. There were no complications occurred such as heterotopic ossification, traumatic arthritis and others. At the latest follow-up, the average angle of elbow flexion was (127.0±5.6)°(120° to 135°); the average extension angle was(4.2±4.5)° (0° to 10°); the average pronation angle of forearm was (86.0±6.1)°(75° to 90°); the average supination angle of forearm was (87.0±6.9)°(80° to 100°). Mayo elbow function score was 80 to 96 points, with an average of 88 points, of which 2 cases got an excellent result, 10 good.</p><p><b>CONCLUSIONS</b>Elbow anterior approach can clearly expose the Regan-Morrey type III coronoid fractures, and mini-locking plate fixation has a satisfactory effect.</p>

Systemic pro-inflammatory response facilitates the development of cerebral edema during short hypoxia.

BACKGROUND: High-altitude cerebral edema (HACE) is the severe type of acute mountain sickness (AMS) and life threatening. A subclinical inflammation has been speculated, but the exact mechanisms underlying the HACE are not fully understood. METHODS: Human volunteers ascended to high altitude (3860 m, 2 days), and rats were exposed to hypoxia in a hypobaric chamber (5000 m, 2 days). Human acute mountain sickness was evaluated by the Lake Louise Score (LLS), and plasma corticotrophin-releasing hormone (CRH) and cytokines TNF-α, IL-1ß, and IL-6 were measured in rats and humans. Subsequently, rats were pre-treated with lipopolysaccharide (LPS, intraperitoneal (ip) 4 mg/kg, 11 h) to induce inflammation prior to 1 h hypoxia (7000 m elevation). TNF-α, IL-1ß, IL-6, nitric oxide (NO), CRH, and aquaporin-4 (AQP4) and their gene expression, Evans blue, Na(+)-K(+)-ATPase activity, p65 translocation, and cell swelling were measured in brain by ELISA, Western blotting, Q-PCR, RT-PCR, immunohistochemistry, and transmission electron micrography. MAPKs, NF-κB pathway, and water permeability of primary astrocytes were demonstrated. All measurements were performed with or without LPS challenge. The release of NO, TNF-α, and IL-6 in cultured primary microglia by CRH stimulation with or without PDTC (NF-κB inhibitor) or CP154,526 (CRHR1 antagonist) were measured. RESULTS: Hypobaric hypoxia enhanced plasma TNF-α, IL-1ß, and IL-6 and CRH levels in human and rats, which positively correlated with AMS. A single LPS injection (ip, 4 mg/kg, 12 h) into rats increased TNF-α and IL-1ß levels in the serum and cortex, and AQP4 and AQP4 mRNA expression in cortex and astrocytes, and astrocyte water permeability but did not cause brain edema. However, LPS treatment 11 h prior to 1 h hypoxia (elevation, 7000 m) challenge caused cerebral edema, which was associated with activation of NF-κB and MAPKs, hypoxia-reduced Na(+)-K(+)-ATPase activity and blood-brain barrier (BBB) disruption. Both LPS and CRH stimulated TNF-α, IL-6, and NO release in cultured rat microglia via NF-κB and cAMP/PKA. CONCLUSIONS: Preexisting systemic inflammation plus a short severe hypoxia elicits cerebral edema through upregulated AQP4 and water permeability by TLR4 and CRH/CRHR1 signaling. This study revealed that both infection and hypoxia can cause inflammatory response in the brain. Systemic inflammation can facilitate onset of hypoxic cerebral edema through interaction of astrocyte and microglia by activation of TLR4 and CRH/CRHR1 signaling. Anti-inflammatory agents and CRHR1 antagonist may be useful for prevention and treatment of AMS and HACE.

Adaptive methylation regulation of p53 pathway in sympatric speciation of blind mole rats, Spalax.

Epigenetic modifications play significant roles in adaptive evolution. The tumor suppressor p53, well known for controlling cell fate and maintaining genomic stability, is much less known as a master gene in environmental adaptation involving methylation modifications. The blind subterranean mole rat Spalax eherenbergi superspecies in Israel consists of four species that speciated peripatrically. Remarkably, the northern Galilee species Spalax galili (2n = 52) underwent adaptive ecological sympatric speciation, caused by the sharply divergent chalk and basalt ecologies. This was demonstrated by mitochondrial and nuclear genomic evidence. Here we show that the expression patterns of the p53 regulatory pathway diversified between the abutting sympatric populations of S. galili in sharply divergent chalk-basalt ecologies. We identified higher methylation on several sites of the p53 promoter in the population living in chalk soil (chalk population). Site mutagenesis showed that methylation on these sites linked to the transcriptional repression of p53 involving Cut-Like Homeobox 1 (Cux1), paired box 4 (Pax 4), Pax 6, and activator protein 1 (AP-1). Diverse expression levels of p53 between the incipiently sympatrically speciating chalk-basalt abutting populations of S. galili selectively affected cell-cycle arrest but not apoptosis. We hypothesize that methylation modification of p53 has adaptively shifted in supervising its target genes during sympatric speciation of S. galili to cope with the contrasting environmental stresses of the abutting divergent chalk-basalt ecologies.