Identification of early Al-responsive genes in rice bean (Vigna umbellata) roots provides new clues to molecular mechanisms of Al toxicity and tolerance.

Significant secretion of citrate from root apex of rice bean (Vigna umbellata) is delayed by several hours under aluminium (Al) stress. However, the molecular basis of regulation of VuMATE1, a gene encoding an Al-activated citrate transporter, remains unclear. In this study, we used suppression subtractive hybridization together with reverse northern blot analysis and qRT-PCR to identify genes with altered transcript levels in the root apex after treatment with low (5 µm) or high (25 µm) concentration of AlCl(3) for a short time (4 h). We found that in addition to VuMATE1, 393 genes showed an early response to Al. Among functionally annotated genes, those related to 'metabolism and energy', 'signal transduction and transcription' and 'transport' was predominantly up-regulated, whereas those associated with 'protein translation, processing and degradation' was predominantly down-regulated. Comparative analysis of transcriptional profiles highlighted candidate genes associated with citrate secretion and revealed several new aspects of the molecular processes underlying Al toxicity and tolerance. Based on the data, it is proposed that metabolic changes represent adaptive mechanisms to Al stress, whereas inhibition of both cell elongation and cell division underlies Al-induced root growth inhibition.

Decreased risk of developing lung cancer in subjects carrying the CLPTM1L rs401681 (G>A) polymorphism: evidence from a meta-analysis.

A genome-wide association study revealed that a single nucleotide polymorphism, CLPTM1L - rs401681 (G>A), located at the 5p15.33 locus was significantly associated with increased risk of various cancers; however, its association with lung cancer is currently inconclusive. In order to explore the relationship between this polymorphism and lung cancer risk more precisely, we performed a meta-analysis of eight eligible studies involving 9935 cases and 11,261 controls. The pooled odds ratio (OR) and the 95% confidence interval (CI) were calculated using a fixed- or random-effect models. Results indicated that this polymorphism was significantly associated with lung cancer risk in all genetic models (GA vs GG: OR = 0.88, 95%CI = 0.83-0.94; AA vs GG: OR = 0.81, 95%CI = 0.70-0.93; AA/GA vs GG: OR = 0.86, 95%CI = 0.81-0.91; AA vs GA/GG: OR = 0.86, 95%CI = 0.76-0.99). An analysis stratified by ethnicity and source of controls revealed a significantly decreased risk among European groups and population-based studies in all genetic models, and among Asian populations only in the dominant model comparison. Additionally, in a subgroup analysis by histology type, the CLPTM1L rs401681 polymorphism was found to significantly decrease the risks of both adenocarcinoma and squamous cell carcinoma of the lung in all genetic models. In conclusion, our study indicated that the CLPTM1L - rs401681 (G>A) polymorphism was significantly associated with decreased lung cancer risk, especially among European populations. Due to some minor limitations, our findings should be confirmed in further studies.

Saccharomyces cerevisiae Live Cells Decreased In vitro Methane Production in Intestinal Content of Pigs.

An in vitro gas production technique was used in this study to elucidate the effect of two strains of active live yeast on methane (CH4) production in the large intestinal content of pigs to provide an insight to whether active live yeast could suppress CH4 production in the hindgut of pigs. Treatments used in this study include blank (no substrate and no live yeast cells), control (no live yeast cells) and yeast (YST) supplementation groups (supplemented with live yeast cells, YST1 or YST2). The yeast cultures contained 1.8×10(10) cells per g, which were added at the rates of 0.2 mg and 0.4 mg per ml of the fermented inoculum. Large intestinal contents were collected from 2 Duroc×Landrace×Yorkshire pigs, mixed with a phosphate buffer (1:2), and incubated anaerobically at 39°C for 24 h using 500 mg substrate (dry matter (DM) basis). Total gas and CH4 production decreased (p<0.05) with supplementation of yeast. The methane production reduction potential (MRP) was calculated by assuming net methane concentration for the control as 100%. The MRP of yeast 2 was more than 25%. Compared with the control group, in vitro DM digestibility (IVDMD) and total volatile fatty acids (VFA) concentration increased (p<0.05) in 0.4 mg/ml YST1 and 0.2 mg/ml YST2 supplementation groups. Proportion of propionate, butyrate and valerate increased (p<0.05), but that of acetate decreased (p<0.05), which led to a decreased (p<0.05) acetate: propionate (A: P) ratio in the both YST2 treatments and the 0.4 mg/ml YST 1 supplementation groups. Hydrogen recovery decreased (p<0.05) with yeast supplementation. Quantity of methanogenic archaea per milliliter of inoculum decreased (p<0.05) with yeast supplementation after 24 h of incubation. Our results suggest that live yeast cells suppressed in vitro CH4 production when inoculated into the large intestinal contents of pigs and shifted the fermentation pattern to favor propionate production together with an increased population of acetogenic bacteria, both of which serve as a competitive pathway for the available H2 resulting in the reduction of methanogenic archaea.

[Plasmids carried by carbapenems-resistant Klebsiella pneumoniae in burn patients and its correlation with strain transmission].

Objective: To explore the carrier status of carbapenems-resistant Klebsiella pneumoniae (CRKP) plasmids in burn patients and analyze the correlation of these plasmids with the transmission of CRKP. Methods: A retrospective observational study was conducted. A total of 26 CRKP strains, which were isolated from the clinic-related samples of 22 burn patients (with 20 males and 2 females, aged (42±16) years) admitted to the First Affiliated Hospital of Army Medical University (the Third Military Medical University) from January to December 2017, were collected and individually numbered. The plasmids of the strains were extracted by alkali lysis. After determination of the plasmid concentration by a nucleic acid concentration detector, the agarose gel electrophoresis was used to visualize the bands, and rough plasmids typing was performed. The plasmid of the smallest numbered CRKP in each plasmid type was transformed into competent Escherichia coli (E. coli) strain Top10 (hereinafter referred to as TOP10 strain). The growth of each transformed strains and a Top10 strain cultivated in ampicillin containing Luria-Bertani (LB) agar medium overnight was observed, and the proportion of successful transformation was calculated. The plasmids from the smallest numbered plasmid carrying CRKP strain of successfully transformed Top10 strains (hereinafter referred to as the smallest successfully transformed strain) and correspondingly numbered CRKP were extracted, and then, the agarose gel electrophoresis was used to visualize the bands. Aforementioned successfully transformed strains and a TOP10 strain were used for the antimicrobial susceptibility testing with 17 antibiotics commonly used in clinic. The plasmid from the smallest successfully transformed strain was sequenced using the next-generation sequencing technology. Bioinformatics analyses such as protein-coding gene prediction and protein sequence alignment were performed successively. The sequence was subsequently named pKP03-NDM1 according to the carrying of drug resistance gene. According to the whole genome sequence of the plasmid carried by the smallest successfully transformed strain, the polymerase chain reaction, agarose gel electrophoresis, and gene sequencing were used to detect the New Delhi metallo-beta lactamase-1 (blaNDM-1) of plasmids in the remaining 25 strains of CRKP. The ST typing in multilocus sequence typing of 26 strains of CRKP was analyzed based on the literature. Results: Plasmids were successfully extracted from 26 CRKP, with mass concentrations ranging from 19.3 to 189.8 ng/µL. Each of the 26 CRKP carrying plasmids showed at least one band longer than 2 500 bp in the agarose gel electrophoresis, which were roughly divided into 6 patterns of A, B, C, D, E, and F. After overnight cultivation, no growth of strains was observed in LB agar medium containing ampicillin inoculated with the TOP10 strain or TOP10 strains transformed by the plasmid of CRKP patterning A, B, D, or E. In contrast, TOP10 strains transformed by the pattern C plasmid from NO.3 CRKP and the pattern F plasmid from NO.15 CRKP resulted in numerous colony growths, and those transformed strains were named as TOP10-pKP03 and TOP10-pKP15, respectively. The proportion of successful transformation was 1/3. The plasmid carried by TOP10-pKP03 showed a single band in the agarose gel electrophoresis, which was the same size as the largest band of the plasmid from NO.3 CRKP. The TOP10 strain was sensitive to the 17 antibiotics commonly used in clinic. TOP10-pKP03 and TOP10-pKP15 were resistant to penicillins, cephalosporins, and carbapenems but remained sensitive to monocyclic ß-lactam, aminoglycosides, quinolones and tigecycline. The full length of the plasmid carried by TOP10-pKP03 was 41 190 bp. In addition to blaNDM-1, this plasmid carried bleMBL, T4SS, bleomycin resistance gene, conjugation transfer elements, and relaxase, etc. The plasmid showed 99% nucleotide identity similarity and the same length to the plasmid pJN24NDM1 extracted from an E. coli isolate JN24. Totally 16 (61.5%) CRKP were confirmed to carrying blaNDM-1 gene, among the ST typing of the 16 strains, 11 strains were ST11, while ST215, ST260, ST395, ST2230, and new ST had 1 strain each. Among the ST typing of 10 blaNDM-1-negative CRKP, 8 strains were ST11, while ST395 and ST2230 had 1 strain each. Conclusions: A blaNDM-1 gene carrying plasmid pKP03-NDM1 was extracted and sequenced from CRKP isolated from burn patients, with a high plasmid carrying rate. Meanwhile, this plasmid may mediate inter-CRKP and CRKP-E. coli horizontal transfer of blaNDM-1, leading to transmission of antimicrobial resistance.

Lysophosphatidic acid (LPA) and endothelial differentiation gene (Edg) receptors in human pancreatic cancer.

Lysophosphatidic acid (LPA) is a naturally occurring phospholipid with diverse effects on various cells, ranging from immediate morphological change to long-lasting cellular function alteration such as induction of stimulation of cell proliferation, survival, drug resistance, and motility. LPA interacts with cells through specific cell surface receptors. LPA1/Edg-2, LPA2/Edg-4, and LPA3/Edg-7 are three most common LPA receptors. Herein we review the roles of LPA and its receptors in the carcinogenesis of human malignancies, with focus on pancreatic cancer.

Long-term survival case of a non-small cell lung cancer bone metastasis patient treated with bone cement, radiation and gefitinib.

OBJECTIVE: We explore the treatment of bone metastases in advanced non-small cell lung cancer (NSCLC). MATERIALS AND METHODS: We reported a 76-year-old female patient, who was diagnosed with NSCLC with bone metastasis eight years ago (stage IVA). Due to unbearable diarrhea, she refused chemotherapy, and we adopted local treatment, including local radiotherapy 50 Gy and bone cement to lumbar spinal metastases, 62 Gy local radiotherapy of primary lung tumor, TKI inhibitor gefitinib and zoledronic acid. RESULTS: She survived more than eight years and is still in follow-up. CONCLUSIONS: The median survival time for NSCLC patients with bone metastases is often less than 1 year. We reported the patient with more than eight years of survival, showed that some special cases can adopt the methods of local treatment including bone cement, treatment benefit patients, radiation therapy and targeted therapy in clinic to expand the survival.

[Epidemiological investigation and analysis of etiological characteristics of infection on 3 067 hospitalized pediatric patients with burns].

Objective: To investigate the epidemiological characteristics and etiological distribution of infection on 3 067 hospitalized pediatric patients with burns, and explore the prevention and treatment strategy of pediatric burns. Methods: A cross-sectional survey was conducted. An analysis was performed on the data of 3 067 hospitalized pediatric patients with burns who met the inclusion criteria and were admitted to the First Affiliated Hospital of Army Medical University (the Third Military Medical University) from January 2012 to December 2020, including gender, age, causative factors, locations and severities of burns, seasons of accidents, and the type, source of tissue or body fluid, and drug resistance of pathogenic bacteria. API bacterial identification batten and automatic microbial identification system were applied for pathogen identification. Drug sensitivities of top 3 consistent ratio pathogen identifed were tested with minimum inhibitory concentration and disk diffusion method. WHONET 5.6 software was applied to analyze the data. Results: There were 3 067 hospitalized pediatric patients with burns, including 1 768 boys and 1 299 girls. The majority of pediatric burn patients were >1 and ≤4 years, accounting for 72.9% (2 236/3 067), and the minority of pediatric burn patients were >8 and ≤12 years, accounting for 4.9% (150/3 067). Moderate burns and severe burns of pediatric burn patients accounted for the majority parts, and the proportions of the two were close. The top cause of pediatric burns was scald, accounting for 81.6% (2504/3 067). Extremities were the most common burn sites in that of entire 3 254. The most pediatric burns occurred in winter, accounting for 29.4% (903/3 067). A total of 1 018 strains of pathogenic bacteria were collected from pediatric burn patients, all of which were non-repeated isolates. The pathogens with top five consistent ratio were Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter baumannii, Enterobacter cloacae, and Escherichia coli, among which Staphylococcus aureus ranked the first every year. The pathogens were mainly isolated from the wound exudate, accounting for 81.34% (828/1 018). Staphylococcus aureus from 2012 to 2020 showed no resistance to vancomycin, linezolid or teicoplanin while Staphylococcus aureus isolated in 2019 was 100% resistant to macrolides, penicillin, aminoglycosides, and quinolones. Pseudomonas aeruginosa was not resistant to polymyxin B. Acinetobacter baumannii showed a high rate of drug resistance to most antibiotics. Conclusions: Among the pediatric burn patients admitted to the First Affiliated Hospital of Army Medical University (the Third Military Medical University) from 2012 to 2020, the majority are male children aged >1 and ≤4 years with moderate burns. Scalds are the leading cause; and extremities are the common burn sites; and the most pediatric burns occurre in winter. Staphylococcus aureus from wound exudate is the primary pathogen of burn wound infections in pediatric patients.

[Work flow of clinical microbiology laboratory in the epidemic of the coronavirus disease 2019].

The burn microbiology laboratory of the author's unit is a level Ⅱ biosafety laboratory, which is mainly responsible for handling clinical microbial samples from our department and other departments in the hospital. Since the outbreak of the coronavirus disease 2019, in order to ensure the normal operation of routine work and the safety of medical staff, the microbiology laboratory has actively adjusted the daily work flow. The detailed work flow is summarized as follows to provide references for the safety protection of peer in clinical microbiology laboratory.

[Analysis of distribution and drug resistance of pathogens isolated from 159 patients with catheter-related bloodstream infection in burn intensive care unit].

Objective: To analyze the distribution and drug resistance of pathogens isolated from patients with catheter-related bloodstream infection (CRBSI) in burn intensive care unit (BICU). Methods: From January 2011 to December 2018, among 2 264 patients who were peripherally inserted central venous catheter at the BICU of the First Affiliated Hospital of Army Medical University (the third Military Medical University), hereinafter referred to as the author's unit, 159 patients were diagnosed CRBSI, including 131 males and 28 females, aged 43 (1, 79) years. The pathogens primarily isolated from peripheral venous blood and central venous catheter blood/anterior central venous catheter specimen of patients with CRBSI were retrospectively analyzed. API bacteria identification kits and automatic microorganism identification instrument were used to identify pathogens. Broth micro-dilution method or Kirby-Bauer paper disk diffusion method was used to detect the drug resistance of the pathogens to 5 antifungal drugs including fluconazole and itraconazole, etc., and 37 antibacterial drugs including tigecycline and imipenem, etc. Modified Hodge test was used to further identify imipenem- and meropenem-resistant Klebsiella pneumonia. D test was used to detect erythromycin-induced clindamycin resistant Staphylococcus aureus. The WHONET 5.6 software was applied to analyze the annual incidence of CRBSI, mortality of patients with CRBSI, incidence of CRBSI cases, distribution of infection site, and duration of catheterization, detection of Gram-negative and Gram-positive bacteria, fungi, methicillin-resistant Staphylococcus aureus (MRSA), and methicillin-sensitive Staphylococcus aureus (MSSA), and drug resistance of fungi and major Gram-negative and Gram-positive bacteria to the commonly used antibiotics in clinic. Results: (1) The incidence of CRBSI was 7.0% (159/2 264) during the eight years, which was slightly higher in 2014 and 2017 with 13.6% (30/221) and 11.1% (24/217) respectively. The mortality rate of patients with CRBSI was 7.5% (12/159). (2) The incidence of CRBSI cases was 14.9% (338/2 264); the main infection site was femoral vein, totally 271 cases (80.2%), and the duration of catheterization of this site was 9 (2, 25) d. (3) During the eight years, totally 543 strains of pathogens were isolated, including 353 (65.0%) strains of Gram-negative bacteria, 140 (25.8%) strains of Gram-positive bacteria, and 50 (9.2%) strains of fungi. The top three isolated pathogens with isolation rate from high to low were Acinetobacter baumannii, Staphylococcus aureus, and Pseudomonas aeruginosa, accounting for 23.2% (126/543), 17.1% (93/543), and 15.7% (85/543), respectively. Fungi were mainly Candida parapsilosis. Among the Staphylococcus aureus, the detection rate of MRSA was 98.9% (92/93), and that of MSSA was 1.1% (1/93). (4) Except for the low drug resistance rates to polymyxin B, minocycline, and tigecycline, the drug resistance rates of Acinetobacter baumannii to the other antibiotics were considerably high (80.1%-100.0%). Pseudomonas aeruginosa was not resistant to polymyxin B but highly resistant to netilmicin (88.7%) and piperacillin (92.6%), with resistance rates to the other antibiotics from 34.5% to 62.7%. Klebsiella pneumoniae was not resistant to tigecycline and lowly resistant to imipenem and meropenem (28.9%, 9 imipenem- and meropenem-resistant strains were further confirmed by modified Hodge test), with resistance rates to the other antibiotics from 40.9% to 95.2%. The resistance rates of MRSA to most antibiotics were higher than those of MSSA. MRSA was not resistant to linezolid, vancomycin, teicoplanin, sulfamethoxazole, or tigecycline. The resistance rates of MRSA to clindamycin and erythromycin were 7.9% and 62.0%, respectively, and those to the other antibiotics were higher than 91.5%. Except for the complete resistance to penicillin G and tetracycline, MSSA was not resistant to the other antibiotics. Thirty-three strains of Staphylococcus aureus showed resistance to erythromycin-induced clindamycin. Fungi was not resistant to amphotericin B, with drug resistance rates to voriconazole, itraconazole, ketoconazole, and fluconazole from 4.2% to 6.2%. Conclusions: The incidence of CRBSI and mortality of patients with CRBSI are high in BICU of the author's unit, and the main infection site is femoral vein. There are various types of pathogens in patients with CRBSI, and most of them are Gram-negative. The top three isolated pathogens are Acinetobacter baumannii, Staphylococcus aureus, and Pseudomonas aeruginosa, accompanying with grim drug resistance phenomenon.

[Analysis of the pathogenic characteristics of fungal bloodstream infection in severe burn patients].

Objective: To retrospectively analyze the diagnosis time, pathogen distribution, and drug resistance of fungal bloodstream infection in severe burn patients. Methods: Blood samples were collected from 55 severe burn patients with fungal bloodstream infection (including 46 males and 9 females, aged 42 (1, 78) years) admitted to the intensive care unit of the Institute of Burn Research of the First Affiliated Hospital of Army Medical University (the Third Military Medical University) from July 2011 to May 2019 for retrospective analysis. Microbial monitoring system was used to cultivate pathogens, API yeast identification kit and Candida chromogenic medium were used to identify pathogens, and Kirby-Bauer paper disk diffusion method was used to detect drug resistance of fungi to fluconazole, amphotericin B, itraconazole, ketoconazole, and voriconazole. The positive rate of blood fungal culture, mortality rate, distribution of local fungal proliferation sites, the diagnosis time distribution of fungal bloodstream infection, the distribution of fungal species, resistance to commonly-used antifungal drugs, and the use of antibiotics were assessed. The WHONET 5.6 software was applied to analyze the distribution and drug resistance of fungi. Results: (1) Totally 4 839 blood samples were collected during the 9 years, and 122 strains of fungi were isolated, with positive rate of 2.52%. The mortality rate was 14.55% (8 patients) in 55 patients. Catheter fungal proliferation ranked the first among 30 cases of local fungal proliferation. (2) The diagnosis time of fungal bloodstream infection mainly distributed in ≤1 week of hospitalization [32.73% (18/55)]. (3) Among the 55 strains of fungi detected, the detection rate of Candida parapsilosis ranked the first (21.82%, 12 strains), Candida glabrata was the second (18.18%, 10 strains), and Candida tropicalis was tied with Candida albicans in the third place (14.55%, 8 strains). All the detected fungi were sensitive to amphotericin B, and the resistance rates to voriconazole, fluconazole, itraconazole, and ketoconazole were between 4.5% and 9.1%. (4) Droad-spectrum antibiotics were used in all the 55 patients, ≥3 kinds of antibiotics were used in 44 patients, and 37 patients used antibacterial drugs ≥7 days. Conclusions: The diagnosis time of fungal bloodstream infection in the 55 severe burn patients was mainly within 1 week of hospitalization. Candida parapsilosis is the most commonly detected fungal species. Catheter fungal proliferation occurs most commonly among the 30 patients with local fungal proliferation. All the detected fungi were sensitive to amphotericin B, with low drug resistance to voriconazole, fluconazole, itraconazole, and ketoconazole. Broad-spectrum antibiotics were overused in the severe burn patients with fungal bloodstream infection.

[Chromosomal aberrations detection in chronic lymphocytic leukemia by conventional cytogenetics using DSP30 and IL-2].

Objective: To study the value of unmethylated cytosine guanine dinucleotide oligodeoxynucleotide (DSP30) and IL-2 in the conventional cytogenetic (CA) detection of the chromosomal aberrations in chronic lymphocytic leukemia (CLL) . Methods: Bone marrow or peripheral blood cells of CLL patients were cultured with DSP30 plus IL-2 for 72 h, following which R-banding analysis was conducted. Fluorescence in situ hybridization (FISH) was performed in 85 patients. CA results were compared with data obtained by FISH. Results: Among 89 CLL patients, the success rate of chromosome analysis was 94.38% (84/89) . Clonal aberrations were detected in 51 patients (51/84, 60.71%) . Of them, 27 (27/51, 52.94%) were complex karyotype. Among 85 CLL patients tested by FISH, chromosomal abnormalities were detected in 74 (74/85, 87.06%) patients, of which 2 (2/74) patients were complex karyotypes, accounting for 2.70%. Of the 85 CLL patients examined by FISH, 50 had abnormal karyotype analysis, 30 had normal karyotype, 5 failed to have chromosome analysis. Among them, 25 cases showed clonal aberrations by FISH assay but normal by CA, and 4 cases were normal by FISH but displayed aberrations in chromosome analysis, and totally 78 (91.76%) cases with abnormality detected by the combination of the two methods. The frequency of 13q- abnormality detected by FISH was significantly higher than that by CA analysis (69.41%vs 16.67%, P<0.001) , while the frequency of 11q-,+12 and 17p- detected by two methods showed no significant difference (P>0.05) . The detection rate of complex abnormalities in conventional karyotype analysis was higher than that in FISH (50.98%vs 2.70%) . In addition, 11 low-risk and 9 intermediate-risk patients according to FISH results showed complex karyotype by cytogenetics, and were classified into high-risk cytogenetic subgroup. Conclusion: DSP30 and IL-2 are effective in improving the detection rate of CA in CLL patients (60.71%) and CA is more effective to detect complex karyotype. However, FISH had a higher overall abnormality detection rate (87.06%) than CA, especially for 13q-. The combination of CA and FISH not only enhanced the detection rate of clonal aberrations to 91.76%, but also provided more precise prognosis stratification for CLL patients, thus to provide more information for clinical implication.

Intravenous fentanyl is exhaled and the concentration fluctuates with time.

Previous studies have reported that fentanyl is eliminated predominantly by hepatic biotransformation, and that some is eliminated unchanged in urine and stools. No reports have described the elimination of fentanyl via the lungs. In this study, exhaled gas samples from eight anaesthetized patients undergoing cardiac surgery were analysed using solid-phase microextraction (SPME) coupled with gas chromatography-mass spectrometry (GC-MS). Results confirmed that fentanyl was exhaled by patients after intravenous administration, that the concentration of exhaled fentanyl fluctuated with time and peak concentrations were reached approximately 15 - 20 min after intravenous fentanyl administration. Thus, in addition to hepatic biotrans formation and elimination via urine and faeces, fentanyl is also eliminated unchanged by the lungs. The potential risk to operating theatre personnel from long-term exposure to low levels of exhaled anaesthetic agents following intravenous administration to patients during surgery warrants further research.

[Epidemiology investigation of carbapenems-resistant Klebsiella pneumoniae in burn care unit].

Objective: To explore the resistance mechanism and gene type of carbapenems-resistant Klebsiella pneumoniae (CRKP) in burn care unit. Methods: A total of 27 CRKP strains were primarily isolated from 22 patients [20 males, 2 females, aged (42±16) years] admitted to burn care unit of Institute of Burn Research of the First Affiliated Hospital of Army Medical University (the Third Military Medical University, hereinafter referred to as our department) from January to December 2017. After identification of bacteria, the months of detection and distribution of sample source were analyzed. Drug resistance tests of 15 antibiotics were conducted. Polymerase chain reaction was used to detect the drug resistant genes. Pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST) were used to analyze the gene type of strains. Results: (1) During the whole year of 2017, CRKP strains were mostly detected in August (8 strains), September (6 strains), and October (5 strains), with no CRKP in January, March, June, November, and December. Five strains from bed units were detected in August (2 strains), September (1 strain), and October (2 strains). (2) Twenty-seven CRKP strains were derived from blood samples (40.7%, 11/27), wound exudate samples (18.5%, 5/27), deep vein catheter samples (11.1%, 3/27), sputum samples (7.4%, 2/27), urine samples (3.7%, 1/27), and bed unit samples (18.5%, 5/27). (3) The 27 CRKP strains were detected with drug-resistance rates of 100.0% to 7 antibiotics including cefoperazone/sulbactam, piperacillin/tazobactam, cefazolin, ceftriaxone, cefepime, ertapenem, and compound sulfamethoxazole, no drug-resistance to tigecycline, with drug-resistance rates higher than 81.0% to the rest 7 antibiotics. (4) Detection rates for resistance gene bla(CTX-M-10), bla(SHV), bla(TEM), bla(CTX-M-14), bla(ACT), and bla(KPC) were all above 92.5%. (5) According to PFGE, the 27 CRKP strains had 6 types (A, A(1), A(2), B, C, and D). Strains of type A were mainly detected in February, May, and September, with detection rate of 37.0% (10/27). Strains of type C were mainly detected in July, August, and October, with detection rate of 48.1% (13/27). Strains of types A(1), A(2), B, and D were scatteredly detected, with detection rate of 3.7% (1/27) respectively. According to MLST, the 27 CRKP strains had 6 STs. ST11 was the most frequent type, accounting for 74.1% (20/27), which was detected in August to October. The detection rate of ST395, ST2230, ST215, ST260, and STnew ranged from 3.7%(1/27) to 7.4%(2/27), and the strains were scatteredly detected. Conclusions: The main source of CRKP from burn care unit of our department was bloodstream. All the CRKP strains showed high drug-resistance rate and complicated resistance mechanism. There were small scale outbreaks caused by CRKP of type A, type C, and ST11, which should be paid more attention to in clinical treatment and infection control.

[Clinical characteristics of burn patients infected with Stenotrophomonas maltophilia and antibiotic resistance of the strains].

Objective: To investigate the clinical characteristics of burn patients infected with Stenotrophomonas maltophilia (SM) and antibiotic resistance of the strains. Methods: Clinical data of burn patients detected with SM, admitted to our unit from July 2011 to July 2017 were retrospectively analyzed. API 20NE bacteria identification panel or fully automated microbial identification instrument was used to identify pathogen. Minimal inhibitory concentration method was used in drug sensitivity test of levofloxacin, compound sulfamethoxazole, minocycline, and cefoperazone/sulbactam. Annual detection of SM, clinical characteristics and prognosis of patients infected with SM, sample source and detection time of SM, detection of the pathogens and antibiotics application of patients before their detection of SM, and drug resistance of SM to the above four antibiotics were analyzed. The results of drug sensitivity test were analyzed by software WHONET 5.5. Results: (1) There were totally 119 patients detected with SM, with 11, 12, 21, 22, 28, 13, and 12 cases from 2011 to 2017, respectively. (2) Among patients infected with SM, there were 86 (72.3%) males and 33 (27.7%) females. Patients aged more than or equal to 65 years accounted for 11.8% (14/119). Patients aged more than or equal to 18 years and less than 65 years accounted for 76.5% (91/119). Patients aged less than 18 years accounted for 11.8% (14/119). Patients with scald were the most common (totally 72 cases, accounted for 60.5%), and patients with total burn area less than or equal to 10% total body surface area were the most common (totally 35 cases, accounted for 29.4%), too. The proportion of patients with history of basic disease was 16.8% (20/119), with tracheotomy of 46.2% (55/119), with deep vein catheterization of 47.9% (57/119), with history of staying in intensive care unit (ICU) of 61.3% (73/119). Seventy-five (63.0%) patients were cured. Twenty-four (20.2%) patients were improved. Fourteen (11.8%) patients gave up treatment. Six (5.0%) patients died. (3) SM detected from wounds exudate of patients occupied the highest proportion (58.0%, 69/119), which was followed by samples of sputum (17.6%, 21/119), blood (14.3%, 17/119), wound tissue (4.2%, 5/119), catheter (4.2%, 5/119), and urine (1.7%, 2/119). The detection time of SM was 10 hours to 71 days post admission, with the average time of 12.7 days. (4) The proportion of patients detected with pathogens before detection of SM was 66.4% (79/119), and Acinetobacter baumannii and Staphylococcus aureus occupied high proportion among the strains. (5) The proportion of patients using antibiotics before detection of SM was 91.6% (109/119), and 44.0% (48/109) patients used 3 kinds of antibiotics or more. The antibiotics were applied for 271 times. The most frequently used antibiotics were glycopeptides antibiotics (63 times), followed by carbapenems antibiotics (61 times). (6) The total sensitivity rates of SM to levofloxacin and minocycline in 7 years were high (91.6% and 99.4%, respectively). The total sensitivity rate of SM to cefoperazone/sulbactam was low (52.5%). The total sensitivity rate of SM to compound sulfamethoxazole was high (77.6%), and the annual sensitivity rate was higher than 90.0% in recent 3 years. Conclusions: Burn patients infecting SM have high rates of tracheotomy and deep vein catheterization, and most of them stay in ICU and use broad-spectrum antibiotics. SM has high sensitivity to levofloxacin, minocycline, and compound sulfamethoxazole.

Mode of action of Saccharomyces cerevisiae in enteric methane mitigation in pigs.

The objectives of this study were to determine the effect and mode of action of Saccharomyces cerevisiae (YST2) on enteric methane (CH4) mitigation in pigs. A total of 12 Duroc×Landrace×Yorkshire male finisher pigs (60±1 kg), housed individually in open-circuit respiration chambers, were randomly assigned to two dietary groups: a basal diet (control); and a basal diet supplemented with 3 g/YST2 (1.8×1010 live cells/g) per kg diet. At the end of 32-day experiment, pigs were sacrificed and redox potential (Eh), pH, volatile fatty acid concentration, densities of methanogens and acetogens, and expression of methyl coenzyme-M reductase subunit A gene were determined in digesta contents from the cecum, colon and rectum. Results showed that S. cerevisiae YST2 decreased (P<0.05) the average daily enteric CH4 production by 25.3%, lowered the pH value from 6.99 to 6.69 in the rectum, and increased the Eh value in cecum and colon by up to -55 mV (P<0.05). Fermentation patterns were also altered by supplementation of YST2 as reflected by the lower acetate, and higher propionate molar proportion in the cecum and colon (P<0.05), resulting in lower acetate : propionate ratio (P<0.05). Moreover, there was a 61% decrease in Methanobrevibacter species in the upper colon (P<0.05) and a 19% increase in the acetogen community in the cecum (P<0.05) of treated pigs. Results of our study concluded that supplementation of S. cerevisiae YST2 at 3 g/kg substantially decreased enteric CH4 production in pigs.

[Analysis of distribution and drug resistance of pathogens from the wounds of 1 310 thermal burn patients].

Objective: To analyze the distribution and drug resistance of pathogens from the wounds of thermal burn patients, so as to provide reliable basis for the rational use of antibiotics and the effective control over nosocomial infection. Methods: Wound samples of 1 310 thermal burn patients admitted into our burn wards from January 2012 to December 2017 were collected and retrospectively analyzed. API bacteria identification panels and automatical bacteria identification equipment were used to identify pathogens. E test was conducted to detect drug resistance of pathogens to vancomycin, tigecycline, and oxacillin. Kirby-Bauer paper disk diffusion method was used to detect drug resistance of pathogens to 31 antibiotics including penicillin G, gentamicin and rifampicin, etc., and drug resistance of fungi to 5 antifungal agents (voriconazole, amphotericin B, fluconazole, itraconazole, and ketoconazole). The WHONET 5.6 software was used to analyze the constituent ratios of Gram-negative bacteria, Gram-positive bacteria, and fungi in each year; the distribution of fungi; the distribution of top 10 bacteria with the highest constituent ratios in each year; the constituent ratios of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA); the drug resistance of top 3 bacteria with the highest constituent ratios to commonly used antibiotics in each year; and the drug resistance of Candida to commonly used antifungal agents. Results: (1) Totally 2 183 strains of pathogens were isolated for the first time, including Gram-negative bacteria 1 194 (54.70%) strains, Gram-positive bacteria 879 (40.27%) strains, and fungi 110 (5.04%) strains. From 2012 to 2016, the constituent ratio of Gram-negative bacteria showed a decreasing trend, while that of Gram-positive bacteria showed an increasing trend year by year; and the constituent ratio of fungi was with a significantly increasing trend from 2016 to 2017. (2) Among all the fungi, the constituent ratio of Candida parapsilosis ranked the first, Aflatoxin ranked the second, Candida albicans and Candida tropicalis both ranked the third. (3) From 2012 to 2017, top 10 bacteria with the highest constituent ratios, from high to low, were Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter baumannii, Enterobacter cloacae, Escherichia coli, Staphylococcus haemolyticus, Klebsiella pneumoniae, Enterococcus faecalis, Aeromonas hydrophila, and Stenotrophomonas maltophilia respectively. The constituent ratio of Staphylococcus aureus ranked the first in each year. The constituent ratio of Pseudomonas aeruginosa was fluctuating but showed a rising trend comprehensively. The constituent ratio of Acinetobacter baumannii went up after decreasing. (4) Among all the Staphylococcus aureus, constituent ratio of MRSA was above 65.00%, while that of MSSA was below 31.00% in each year. (5) From 2012 to 2017, Staphylococcus aureus resistant to vancomycin, linezolid, or teicoplanin was not detected; the drug-resistant rates of MRSA to penicillin G, oxacillin, gentamicin, rifampicin, tetracycline, ciprofloxacin, ofloxacin, and levofloxacin were above or equal to 80.0% in each year; the drug-resistant rates of Staphylococcus aureus to clindamycin and erythrocin showed an obviously increasing trend, the drug-resistant rates of Staphylococcus aureus to moxifloxacin and queenoputin/daputin in 2017 were higher than those in 2016, while the drug-resistant rates of Staphylococcus aureus to the other 14 antibiotics showed no significant change in trend. From 2012 to 2017, Acinetobacter baumannii was sensitive to polymyxin B and tigecycline; the drug-resistant rate of Acinetobacter baumannii to ceftriaxone was relatively high; the drug-resistant rates of Acinetobacter baumannii to levofloxacin, minocycline, and tetracycline were decreasing while those to the other 14 antibiotics went up after decreasing. From 2012 to 2017, Pseudomonas aeruginosa wasn't resistant to polymyxin B, and its drug-resistant rates to the other 14 antibiotics showed decreasing trends. (6) The drug-resistant rates of Candida albicans to voriconazole, amphotericin B, fluconazole, itraconazole, and ketoconazole were all zero. The drug-resistant rates of non-Candida albicans to voriconazole, fluconazole, itraconazole, and ketoconazole were higher than those of Candida albicans. Conclusions: Among the pathogens from the wounds of thermal burn patients, Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii had the top 3 constituent ratios; the constituent ratio of non-Candida albicans was obviously higher than that of Candida albicans. The high drug resistance rates of Staphylococcus aureus and Acinetobacter baumanni require more attention from clinicians and the local hospital's infection control department.

[Application of recombinase polymerase amplification in the detection of Pseudomonas aeruginosa].

Objective: To establish an optimized method of recombinase polymerase amplification (RPA) to rapidly detect Pseudomonas aeruginosa in clinic. Methods: (1) The DNA templates of one standard Pseudomonas aeruginosa strain was extracted and detected by polymerase chain reaction (PCR), real-time fluorescence quantitative PCR and RPA. Time of sample loading, time of amplification, and time of detection of the three methods were recorded. (2) One standard Pseudomonas aeruginosa strain was diluted in 7 concentrations of 1×10(7,) 1×10(6,) 1×10(5,) 1×10(4,) 1×10(3,) 1×10(2,) and 1×10(1) colony forming unit (CFU)/mL after recovery and cultivation. The DNA templates of Pseudomonas aeruginosa and negative control strain Pseudomonas putida were extracted and detected by PCR, real-time fluorescence quantitative PCR, and RPA separately. The sensitivity of the three methods in detecting Pseudomonas aeruginosa was analyzed. (3) The DNA templates of one standard Pseudomonas aeruginosa strain and four negative control strains (Staphylococcus aureus, Acinetobacter baumanii, Candida albicans, and Pseudomonas putida) were extracted separately, and then they were detected by PCR, real-time fluorescence quantitative PCR, and RPA. The specificity of the three methods in detecting Pseudomonas aeruginosa was analyzed. (4) The DNA templates of 28 clinical strains of Pseudomonas aeruginosa preserved in glycerin, 1 clinical strain of which was taken by cotton swab, and negative control strain Pseudomonas putida were extracted separately, and then they were detected by RPA. Positive amplification signals of the clinical strains were observed, and the detection rate was calculated. All experiments were repeated for 3 times. Sensitivity results were analyzed by GraphPad Prism 5.01 statistical software. Results: (1) The loading time of RPA, PCR, and real-time fluorescence quantitative PCR for detecting Pseudomonas aeruginosa were all 20 minutes. In PCR, time of amplification was 98 minutes, time of gel detection was 20 minutes, and the total time was 138 minutes. In real-time fluorescence quantitative PCR, amplification and detection could be completed simultaneously, which took 90 minutes, and the total time was 110 minutes. In RPA, amplification and detection could also be completed simultaneously, which took 15 minutes, and the total time was 35 minutes. (2) Pseudomonas putida did not show positive amplification signals or gel positive results in any of the three detection methods. The detection limit of Pseudomonas aeruginosa in real-time fluorescence quantitative PCR and PCR was 1×10(1) CFU/mL, and that of Pseudomonas aeruginosa in RPA was 1×10(2) CFU/mL. In RPA and real-time fluorescence quantitative PCR, the higher the concentration of Pseudomonas aeruginosa, the shorter threshold time and smaller the number of cycles, namely shorter time for detecting the positive amplified signal. In real-time fluorescence quantitative PCR, all positive amplification signal could be detected when the concentration of Pseudomonas aeruginosa was 1×10(1)-1×10(7) CFU/mL. In RPA, the detection rate of positive amplification signal was 0 when the concentration of Pseudomonas aeruginosa was 1×10(1) CFU/mL, while the detection rate of positive amplification signal was 67% when the concentration of Pseudomonas aeruginosa was 1×10(2) CFU/mL, and the detection rate of positive amplification signal was 100% when the concentration of Pseudomonas aeruginosa was 1×10(3)-1×10(7) CFU/mL. (3) In RPA, PCR, and real-time fluorescence quantitative PCR, Pseudomonas aeruginosa showed positive amplification signals and gel positive results, but there were no positive amplification signals or gel positive results in four negative control strains of Acinetobacter baumannii, Staphylococcus aureus, Candida albicans, and Pseudomonas putida. (4) In RPA, 28 clinical strains of Pseudomonas aeruginosa preserved in glycerin and 1 clinical strain of Pseudomonas aeruginosa taken by cotton swab showed positive amplification signals, while Pseudomonas putida did not show positive amplification signal. The detection rate of positive amplification signal of 29 clinical strains of Pseudomonas aeruginosa in RPA was 100%. Conclusions: The established optimized RPA technology for fast detection of Pseudomonas aeruginosa requires shorter time, with high sensitivity and specificity. It was of great value in fast detection of Pseudomonas aeruginosa infection in clinic.

[Clinical significance of expressions of EVI1 and BRE genes in 47 acute leukemia patients with MLL rearrangement].

Objective: To investigate the overexpression frequencies of BRE and EVI1, the correlation between BRE and EVI1 expressions and their possible clinical implications in 11q23/MLL rearrangement acute leukemia. Methods: Cytogenetic examination of bone marrow cells was performed by short-term culture method. R-banding technique was used for karyotype analysis. 47 patients were detected by interphase fluorescence in situ hybridization (FISH) with dual-color break apart MLL probe. The expressions of EVI1 and BRE genes were detected by real time quantitative reverse transcription polymerase chain reaction (RQ-PCR) . The correlation and prognostic significance were statistically tested. Results: 11q23/MLL rearrangements were confirmed by karyotyping and FISH, respectively in 47 patients. According to immunophenotypic analyses of 37 patients, 5 patients showed positive for CD19, CD79a or CD10, 1 for CD7; the others for CD33, CD13, CD14 and CD15, and 16 of them for CD34. Of the 47 patients, 18 patients showed EVI1 overexpression and most of them presented with t (6;11) and M(4)/M(5). The EVI1 expression was high in t (6;11) or t (9;11) subgroup comparable with levels observed in normal subgroup (P=0.038, 0.022, respectively) . 15 patients showed high BRE expression, and most of them presented with t (9;11) and M(4)/M(5). High BRE expression was found in t (4;11) , t (6;11) , t (9;11) and t (11;19) subgroups, respectively by comparing with normal subgroup. The BRE expression was higher in t (4;11) (P=0.004) or t (9;11) (P=0.012) subgroup than in t (6;11) subgroup. Patients with EVI1 overexpression had a short survival compared with those with low EVI1 expression (P=0.049) and it also did in t (9;11) subgroup (P=0.024) . Patients with t (9;11) and high BRE expression had a long survival compared with those with t (9;11) and low BRE expression (P=0.024) . Conclusion: The EVI1 overexpression was significantly frequent in acute leukemia patients with 11q23/MLL rearranged, especially within t (6;11) subgroup and M(4)/M(5), which was associated with an inferior outcome. High BRE expression was observed frequently in 11q23/MLL-rearranged acute leukemia especially within t (9;11) subgroup and M(5).

Involvement of ghrelin in nucleus tractus solitaries on gastric signal afferent and gastric motility in cisplatin-treated rats.

OBJECTIVE: Ghrelin had been known to promote gastric motility in human and animals previously. We aim to investigate the role of ghrelin in chemotherapy-induced nausea and vomiting. MATERIALS AND METHODS: In the present study, we observed the changes in food intake, kaolin consumption, body weight, plasma ghrelin concentration and expression of ghrelin and its receptor GHS-R1a in the stomach and nucleus tractus solitaries (NTS) in cisplatin-treated rats, and the effects of ghrelin microinjected into NTS on the discharge activity of gastric distension (GD) responsive neurons and gastric motility were also observed. RESULTS: Cisplatin induced the decrease in food intake and the increase in kaolin consumption of rats. In addition, mRNA expression of GHS-R1a in the stomach and NTS increased significantly after cisplatin treatment. The discharge activity of GD excited (GD-E) and GD inhibited (GD-I) neurons in cisplatin-treated rats was weaker than that of saline treatment, while ghrelin administration into NTS excited most of GD-E and GD-I neurons. Cisplatin induced the decrease in gastric contraction while ghrelin administrated into NTS promoted the gastric motility significantly. However, the amplitude and frequency of gastric contraction promoted by ghrelin in NTS of cisplatin-treated rats were lower than that of saline treated rats. The effects of ghrelin could be completely blocked by its receptor antagonist BIM28163. CONCLUSIONS: These results indicated that ghrelin in the NTS might participate in the regulation of GD-neurons and gastric motility via its receptor in cisplatin-treated rats.

A rare occurrence of intramasseteric schwannoma - case report and literature review.

A schwannoma is a benign, solitary, well-defined, painless, slowly-enlarging nerve sheath tumor, composed of Schwann cells. Intramasseteric localization is very unusual. We report the case of a 33-year-old male who developed an intramasseteric schwannoma. Tumor could be completely removed under general anesthesia. Histopathological examination made the diagnosis of intramasseteric schwannoma through the presence of Antoni A areas and Verocay bodies. The diagnosis of schwannoma should be taken into consideration in case of parotideomasseteric tumors.