Transcriptome analysis of Kluyveromyces marxianus under succinic acid stress and development of robust strains.

Kluyveromyces marxianus has become an attractive non-conventional yeast cell factory due to its advantageous properties such as high thermal tolerance and rapid growth. Succinic acid (SA) is an important platform molecule that has been applied in various industries such as food, material, cosmetics, and pharmaceuticals. SA bioproduction may be compromised by its toxicity. Besides, metabolite-responsive promoters are known to be important for dynamic control of gene transcription. Therefore, studies on global gene transcription under various SA concentrations are of great importance. Here, comparative transcriptome changes of K. marxianus exposed to various concentrations of SA were analyzed. Enrichment and analysis of gene clusters revealed repression of the tricarboxylic acid cycle and glyoxylate cycle, also activation of the glycolysis pathway and genes related to ergosterol synthesis. Based on the analyses, potential SA-responsive promoters were investigated, among which the promoter strength of IMTCP2 and KLMA_50231 increased 43.4% and 154.7% in response to 15 g/L SA. In addition, overexpression of the transcription factors Gcr1, Upc2, and Ndt80 significantly increased growth under SA stress. Our results benefit understanding SA toxicity mechanisms and the development of robust yeast for organic acid production. KEY POINTS: • Global gene transcription of K. marxianus is changed by succinic acid (SA) • Promoter activities of IMTCP2 and KLMA_50123 are regulated by SA • Overexpression of Gcr1, Upc2, and Ndt80 enhanced SA tolerance.

Serum uric acid and outcome in hospitalized elderly patients with chronic heart failure through the whole spectrum of ejection fraction phenotypes.

INTRODUCTION: Elevated serum uric acid (SUA) levels have been associated with poor outcome in patients with heart failure (HF). Uric acid is associated with inflammation and microvascular dysfunction, which may differentially affect left ventricular ejection fraction (EF) phenotypes. We aimed to identify the role of SUA across EF phenotypes in hospitalized elderly patients with chronic HF. METHODS: We analyzed 1355 elderly patients who were diagnosed with chronic HF. All patients had SUA levels measured within the first 24 h following admission. Patients with left ventricle EF were categorized as having HF with reduced EF (HFrEF, EF < 40%), HF with mid-range EF (HFmrEF, 40%≦LVEF ≦ 49%) or HF with preserved EF (HFpEF, LVEF ≥ 50%). Endpoints were cardiovascular death, HF rehospitalization, and their composite. The median follow-up period was 18 months. RESULTS: Compared with the lowest SUA quartile, the highest SUA quartile was significantly associated with the endpoints (adjusted HR: 2.404, 95% CI: 1.178-4.906, P = 0.016; HR: 1.418, 95% CI: 1.021-1.971, P = 0.037; HR: 1.439, 95% CI: 1.049-1.972, P = 0.024, respectively). After model adjustment, a significant association of SUA with cardiovascular death and the composite endpoint persisted among HFrEF and HFmrEF patients in the highest SUA quartile (P < 0.05 for all). CONCLUSIONS: In hospitalized elderly patients with chronic HF, SUA is an independent predictor of adverse outcomes, which can be seen in HFrEF and HFmrEF patients.

Velocity mismatch effect on the dynamic MTF of a TDI image sensor.

By analyzing the process of time delay integration dynamic imaging, we establish a model of velocity mismatch. Based on this model, we analyze the influence of different factors on the dynamic imaging process, and a modulation transfer function (MTF) is used to evaluate imaging quality. According to the simulation, the velocity mismatch and scan stage are the main factors for image quality. The MTF of the image sensor decreases with the velocity mismatch, and the scan stage increases. In addition, an image with higher contrast can be obtained in a short integration time. However, a shorter integration time leads to insufficient sampling. Furthermore, we establish a dynamic MTF testing system, and evaluate the experiment at different imaging modes. Through data comparison, the experimental data are consistent with theoretical data.

Novel plasmid-mediated colistin resistance gene mcr-7.1 in Klebsiella pneumoniae.

Objectives: To identify a novel plasmid-mediated colistin resistance gene in Klebsiella pneumoniae isolated from chickens in China. Methods: WGS was used to identify a novel colistin resistance gene. The transferability of plasmids carrying mcr-7.1 was investigated by conjugation experiments. The expression of the mcr-7.1 gene was examined using an expression vector. Results: A novel plasmid-mediated colistin resistance gene mcr-7.1, sharing 70% amino acid identity with the mcr-3 gene, was identified in three K. pneumoniae strains isolated from chickens in China. The mcr-7.1 gene was found in an IncI2-type plasmid (pSC20141012) that co-harboured the blaCTX-M-55 gene in one isolate. pSC20141012 can be transferred from K. pneumoniae SC20141012 to Escherichia coli J53Azr, exhibiting a ≥8-fold increase in colistin MIC compared with the recipient E. coli J53Azr. Conclusions: We identified a novel plasmid-mediated colistin resistance gene named mcr-7.1 in K. pneumoniae in China. The prevalence of mcr-7.1 in various species of human and animal origin needs to be investigated immediately.

Colistin Resistance Gene mcr-1 and Its Variant in Escherichia coli Isolates from Chickens in China.

The mcr-1 gene was detected in 5.11% (58/1136) of Escherichia coli isolates of chicken origin from 13 provinces in China. A novel mcr-1 variant, named mcr-1.3, encoding an Ile-to-Val functional variant of MCR-1 was identified in a sequence type 155 (ST155) strain. An mcr-1.3-containing IncI2 plasmid, pHeN867 (60,757 bp), was identified. The transfer of pHeN867 led to a 32-fold increase in the MIC of colistin in the recipient, exhibiting an effect on colistin resistance that was similar to that of mcr-1.

Characterization of pHeBE7, an IncFII-type virulence-resistance plasmid carrying blaCTX-M-98b, blaTEM-1, and rmtB genes, detected in Escherichia coli from a chicken isolate in China.

Recently, a novel variant of the CTX-M enzyme, CTX-M-98, was detected in Escherichia coli isolates from food animals. However, few plasmids carrying blaCTX-M-98 have been fully characterized. In this study, we sequenced the complete pHeBE7 plasmid, an 86,015-bp plasmid that contains the blaCTX-M-98b, blaTEM-1, rmtB, and traT genes, using whole-genome sequencing. The backbone of pHeBE7 shows a high similarity (>99%) to pMC-NDM, which carries the blaNDM-1 gene, however its mosaic regions remain relatively unique among sequenced plasmids. We discovered that a typical ISEcp1-blaCTX-M-IS903 element in the mosaic region harbors the blaCTX-M-98b gene. Conjugation and growth competition assays indicate that pHeBE7 can be easily transmitted and that it confers a limited fitness cost to the recipient cell. The genetic characterization of pHeBE7 may improve our knowledge of how antibiotic resistance disseminates in enterobacteria.

Development and validation of clinical risk score to predict the cardiac rupture in patients with STEMI.

BACKGROUND: Cardiac rupture (CR) is a fatal complication of ST-elevation myocardial infarction (STEMI) with poor prognosis. The aim of this study was to develop and validate practical risk score to predict the CR after STEMI. METHODS: A total of 11,234 STEMI patients from 7 centers in China were enrolled in our study, we firstly developed a simplified fast-track CR risk model from 7455 STEMI patients, and then prospectively validated the CR risk model using receiver-operating characteristic (ROC) curves by the other 3779 consecutive STEMI patients. This trial is registered with ClinicalTrials.gov, number NCT02484326. RESULTS: The incidence of CR was 2.12% (238/11,234), and the thirty-day mortality in CR patients was 86%. We developed a risk model which had 7 independent baseline clinical predictors (female sex, advanced age, anterior myocardial infarction, delayed admission, heart rate, elevated white blood cell count and anemia). The CR risk score system differentiated STEMI patients with incidence of CR ranging from 0.2% to 13%. The risk score system demonstrated good predictive value with area under the ROC of 0.78 (95% CI 0.73-0.84) in validation cohort. Primary percutaneous coronary intervention decreased the incidence of CR in high risk group (3.9% vs. 6.2%, p<0.05) and very high risk group (8.0% vs. 15.2%, p<0.05). CONCLUSIONS: A simple risk score system based on 7 baseline clinical variables could identify patients with high risk of CR, for whom appropriate treatment strategies can be implemented.

Nitrogen removal and its relationship with the nitrogen-cycle genes and microorganisms in the horizontal subsurface flow constructed wetlands with different design parameters.

This study aims to investigate nitrogen removal and its relationship with the nitrogen-cycle genes and microorganisms in the horizontal subsurface flow constructed wetlands (CWs) with different design parameters. Twelve mesocosm-scale CWs with four substrates and three hydraulic loading rates were set up in the outdoor. The result showed the CWs with zeolite as substrate and HLR of 20 cm/d were selected as the best choice for the TN and NH3-N removal. It was found that the single-stage mesocosm-scale CWs were incapable to achieve high removals of TN and NH3-N due to inefficient nitrification process in the systems. This was demonstrated by the lower abundance of the nitrification genes (AOA and AOB) than the denitrification genes (nirK and nirS), and the less diverse nitrification microorganisms than the denitrification microorganisms in the CWs. The results also show that microorganism community structure including nitrogen-cycle microorganisms in the constructed wetland systems was affected by the design parameters especially the substrate type. These findings show that nitrification is a limiting factor for the nitrogen removal by CWs.

Characterization of SXT/R391 Integrative and Conjugative Elements in Proteus mirabilis Isolates from Food-Producing Animals in China.

SXT/R391 integrative and conjugative elements (ICEs) were detected in 8 out of 125 Proteus mirabilis isolates from food-producing animals in China. Whole-genome sequencing revealed that seven ICEs were identical to ICEPmiJpn1, carrying the cephalosporinase gene blaCMY-2. Another one, designated ICEPmiChn1, carried five resistance genes. All eight ICEs could be transferred to Escherichia coli via conjugation. The results highlight the idea that animal farms are important reservoir of the SXT/R391 ICE-containing P. mirabilis.

[Standardized investigation on medicinal nature of ethnomedicine].

This article recorded the analysis and comparison between the medicinal nature theory of traditional Chinese medicine(TCM) and ethnomedicine(EM). The vocabulary of "medicinal nature" was suggested to indicate the properties of ethnomedicine. Based on the influence of TCM medicinal nature theory on EM in China, the application of medicinal nature theory in EM was divided into 3 classes, and the standardizing principles for EM medicinal nature were proposed. It was suggested that medicinal quality, flavor, tendency, tropism, degree and efficiency can be used for the classification standard for EM medicinal nature.

Two novel Salmonella genomic island 1 variants in Proteus mirabilis isolates from swine farms in China.

Four different Salmonella genomic island 1 (SGI1) variants, including two novel variants, were characterized in one Salmonella enterica serovar Rissen sequence type ST1917 isolate and three Proteus mirabilis isolates from swine farms in China. One novel variant was derived from SGI1-B with the backbone gene S021 disrupted by a 12.72-kb IS26 composite transposon containing the dfrA17-aadA5 cassettes and macrolide inactivation gene cluster mphA-mrx-mphR. The other one was an integron-free SGI1 and contained a 183-bp truncated S025 next to IS6100 and S044.

Intraoperative anthropometric measurements of tibial morphology: comparisons with the dimensions of current tibial implants.

PURPOSES: This study analyzed morphological differences in the resected proximal tibial surfaces of Chinese males and females undergoing total knee arthroplasty (TKA) and compared the measurements with the dimensions of five currently used tibial implants. METHODS: The mediolateral (ML), middle anteroposterior (AP), medial anteroposterior (MAP), and lateral anteroposterior (LAP) dimensions of the resected tibial surfaces of 976 Chinese TKA knees (177 male, 799 female) were measured. The ML/AP ratio of every knee was calculated. These morphological data were compared with the dimensions of five currently used tibial implants. RESULTS: The ML, AP, MAP, and LAP dimensions of the resected proximal tibias showed significant differences according to gender. Compared with currently used tibial implants, the smaller implants showed tibial ML undersizing and the larger implants showed tibial ML overhang. The ML/AP aspect ratio progressively decreased with increasing AP dimension in the resected proximal tibias, which contrasts with the relatively constant or increased (NexGen) aspect ratio in currently used tibial implants. Males showed a higher ML/AP aspect ratio than females for a given AP dimension. This indicates that for an implant with a given AP dimension, the tibial ML dimension tends to be undersized in males and to overhang in females. CONCLUSION: The results of this study may provide fundamental data for designing suitable tibial implants for use in the Chinese population, especially for design of gender-specific prostheses. LEVEL OF EVIDENCE: II.

[Effect of diets with different fat levels on the body size and development of Lucilia sericata].

OBJECTIVE: To observe the effect of diets with different fat levels on the body size and development of Lucilia sericata. METHODS: Under the constant temperature of 28 degrees C, the larvae were reared on the diets containing 0% (G0), 10% (G1), 30% (G3), 50% (G5) and 80% (G8) fat tissues (fat/muscle ratio), respectively. Length and weight of larvae and pupae were measured at 12 h interval since 16 h after eclosion. Length of inter-medial cross vein (m-m) of adult left wing was measured. 10 samples were collected in each group. The developmental duration time, mortality and sex ratios of adults were recorded. RESULTS: The mean maximal larval length [(13.3 +/- 1.2), (12.0 +/- 1.1), (10.2 +/- 0.9) and (8.8 +/- 0.8) mm, respectively] and mean maximal larval weight [(72.8 +/- 6.1), (62.2 +/- 5.7), (47.2 +/- 4.3), and (34.9 +/- 5.7) mg] in G1, G3, G5 and G8 groups were significantly less than that of the G0 group [(14.8 +/- 1.3) mm and (80.4 +/- 8.1) mg](P < 0.01). The body size of pupae and adults was also significantly less than that of G0 group (P < 0.01). The total duration time of G5 and G8 groups [(293.3 +/- 22.2) and (285.2 +/- 24.6) h] were significantly shorter than that of G0 group [(312.8 +/- 20.1)h] (P < 0.01). The mortality of larvae [(32.6 +/- 5.6)% and (44.3 +/- 7.7)%] and pupae [(28.6 +/- 5.5)% and (43.5 +/- 6.2)%] of G5 and G8 group were also significantly higher than that of G0 group [(5.7 +/- 3.3)% and (4.5 +/- 1.9)%] (P < 0.01). There was no significant difference in sex ratio among the 5 groups (P > 0.05). CONCLUSION: The body size of larvae, pupae and adults of Lucilia sericata is smaller, the development time is shorter and mortality is higher when the food substrate contains more fat tissues.

Solid basal adenoid cystic carcinoma of the breast: A case report and literature review.

RATIONALE: Adenoid cystic carcinoma (AdCC) is a rare malignancy of the breast with a low Ki-67 index and good prognosis. Owing to the rarity of breast AdCC, the misdiagnosis rate is as high as 50%, and there is no consensus or recognized guidelines for the treatment of this disease. Therefore, it is necessary to conduct a detailed clinical and pathological analysis in combination with a literature review to improve our understanding, diagnosis, and treatment of the disease. METHODS: A 68-year-old woman sought medical attention due to a recently increasing mass in the breast. The left breast mass was 1.3 cm × 1 cm in size. We analyzed the morphology, immunohistochemistry, and molecular characteristics of the tumor removed by surgery, and reviewed relevant literature. DIAGNOSES: Solid basal AdCC of the breast. INTERVENTIONS: We performed biopsy, immunohistochemistry and molecular testing on surgical resection specimens. OUTCOMES: Combining morphological and immunohistochemical features, it is consistent with solid basal AdCC of the breast, and Fish detected MYB gene break. LESSONS: Due to the high misdiagnosis rate of AdCC, accurate histopathological diagnosis is particularly important. At present, breast conserving surgery and local tumor resection are mainly used for the treatment of breast AdCC, and postoperative adjuvant radiotherapy is feasible.

The displaced polymer clamp entered the duodenum causing an abscess: A case report.

RATIONALE: The Hem-o-lok clip, made from a nonabsorbable polymer, and its predecessor the metal ligation clip have been used widely for laparoscopic or robot-assisted surgery to ligate the cystic duct after a cholecystectomy, to ligate the appendix after an appendectomy, or control hemorrhage or on occasion to occlude a fistula or enterotomy. Displacement of these ligation clips to distant sites is an extremely rare complication in clinical practice. PATIENT CONCERNS: The patient is a 67-year old female who sought medical attention for 3 days due to worsening intermittent upper abdominal pain and poor appetite. Gastroscopy showed both an ulcer and the presence of a foreign object embedded in the anterior wall of the duodenal bulb, consistent with what looked like a polymer-based ligation clip. After removal of the foreign body, which turned out to be a remnant of the polymer clip, no further pus was seen, but fresh granulomatous tissues were seen at the base. DIAGNOSES: a polymer-based clip-induced duodenal bulb erosion with a local contained enterically draining abscess. INTERVENTIONS: The patient recovered after removing foreign bodies under gastroscopy and receiving anti infection treatment. OUTCOMES: The patient recovered after removing foreign bodies under gastroscopy and receiving anti infection treatment. LESSONS: In laparoscopic cholecystectomy, attention should be paid to the correct surgical techniques, possibly by decreasing the number of such clips used or considering use of absorbable clips, ligature wires, ligation with absorbable suture material, or ultrasonic resection, all of which can be used for clipless cholecystectomy.

Establishment of a blunt impact-induced brain injury model in rabbits.

OBJECTIVE: To establish an animal model to replicate the blunt impact brain injury in forensic medicine. METHODS: Twenty-four New Zealand white rabbits were randomly divided into control group (n equal to 4), minor injury group (n equal to 10) and severe injury group (n equal to 10). Based on the BIM-II Horizontal Bio-impact Machine, self-designed iron bar was used to produce blunt brain injury. Two rabbits from each injury group were randomly selected to monitor the change of intracranial pressure (ICP) during the impacting process by pressure microsensors. Six hours after injury, all the rabbits were dissected to observe the injury morphology and underwent routine pathological examination. RESULTS: Varying degrees of nervous system positive signs were observed in all the injured rabbits. Within 6 hours, the mortality rate was 1/10 in the minor injury group and 6/10 in the severe injury group. Morphological changes consisted of different levels of scalp hematoma, skull fracture, epidural hematoma, subdural hematoma, subarachnoid hemo- rrhage and brain injury. At the moment of hitting, the ICP was greater in severe injury group than in mild injury group; and within the same group, the impact side showed positive pressure while the opposite side showed negative pressure. CONCLUSIONS: Under the rigidly-controlled experimental condition, this animal model has a good reproducibility and stable results. Meanwhile, it is able to simulate the morphology of iron strike-induced injury, thus can be used to study the mechanism of blunt head injury in forensic medicine.

Changes in the incidence and prevalence of ischemic stroke and associations with natural disasters: an ecological study in 193 countries.

Epidemiological studies have indicated that natural disasters have important impacts on ischemic stroke. This study determined the associations between natural disasters and the incidence and prevalence of ischemic stroke at the global level. A 28-year ecological trend study was performed to estimate worldwide changes in the incidence and prevalence of ischemic stroke and their associations with natural disasters by analyzing data from 193 countries. Quantum geographic information system-based visualization and multivariable linear regression were used. Changes in the incidence and prevalence of ischemic stroke, as well as disaster occurrence, varied among the different regions over the past 28 years (p < 0.001). Multiple linear regression revealed an independent and positive association between disaster occurrence and the incidence of ischemic stroke in males, females and both sexes combined (standardized coefficients = 0.515, 0.470 and 0.483, p < 0.001); similar associations were found for the prevalence of ischemic stroke (standardized coefficients = 0.471, 0.417 and 0.438, p < 0.001). The incidence and prevalence of ischemic stroke changed significantly at the global level and were independently associated with natural disasters. Both males and females show common but different vulnerabilities to natural disasters. This evidence supports policy making and resource allocation for disaster response and disease burden reduction.

[Optimization of Tidal-Combined Flow Constructed Wetland System and Its Removal Effect on Antibiotic Resistance Genes].

Antibiotic resistance genes (ARGs) pose a serious threat to environmental biology and public health, along with the discharge and spread of wastewater. The advanced treatment of ARGs in wastewater therefore deserves special attention. In our previous study, we found that tidal flow constructed wetlands can effectively remove multiple ammonia from wastewater. In this study, we further optimized tidal flow constructed wetland systems by adding baffles and cultivating plants; we investigated the influence of process optimization on the removal of ARGs and the influence of functional microorganism distribution on nitrogen removal. The results show that the addition of baffles and plants can effectively improve the removal efficiency of ARGs, with the maximum removal rate of 21 resistance genes, in 7 categories, reaching 83.82%-100.0% with the simultaneous addition of baffles and plants. These removal rates were significantly higher than the increase resulting from a single baffle or plant group. From the comparison of the absolute abundance of ARGs in the substrate and plants, it is clear that the baffles can promote the enrichment of ARGs in the wetland substrate, while uptake by plants is also a way of removing ARGs. Combined with the results of nitrogen-cycle functional gene sequencing, system optimization can increase the diversity and richness of nitrification and denitrification functional microorganisms in the substrate, which is consistent with the higher removal rate of nitrification and total nitrogen in wastewater.