Characterization of an NDM-1-Producing Citrobacter koseri Isolate from China.

Purpose: The continuous rise in carbapenemase-producing Enterobacteriaceae infections is a major public health concern. However, there is limited information available on New Delhi metallo-ß-lactamase-1 (NDM-1) producing Citrobacter koseri. In this study, we isolated a blaNDM-1-carrying C. koseri from a stool sample of an inpatient. Our aim was to investigate the phenotypic and genomic features of this clinically derived carbapenem-resistant C. koseri isolate and to characterize the transmission pattern of the IncFII/IncN plasmid that carries the blaNDM-1 gene. Methods and Results: S1-PFGE, Southern blot and conjugation assay confirmed the presence of blaNDM-1 gene in a conjugative plasmid. C. koseri L2395 and transconjugant L2395-EC600 strains showed similar resistance spectrum. Whole-genome analysis revealed that pL2395_NDM is an IncFII/IncN plasmid with a length of 67,839 bp. Moreover, blaNDM-1 gene was found encoded in the ISKpn19-blaNDM-1-ble-tnpF-dsbD-cutA-ISKpn19 cassette array. Phylogenetic analysis revealed that strain L2395 was close to an IMP-4-bearing C. koseri from Australia. Conclusion: Ongoing surveillance will be essential to control and prevent the spread of carbapenem-resistant Citrobacter spp. in the future.

[Expression level of cytokines in patients with sepsis and its effect on prognosis].

OBJECTIVE: To observe the expression level of cytokines in patients with sepsis and its effect on prognosis. METHODS: The clinical data of sepsis patients admitted to the intensive care unit (ICU) of the First Affiliated Hospital of Zhengzhou University from January 2020 to December 2022 were analyzed retrospectively, including gender, age, and acute physiology and chronic health evaluation II (APACHE II), blood routine, procalcitonin (PCT), C-reactive protein (CRP), and cytokines levels [interleukins (IL-2, IL-4, IL-6, IL-10, IL-17), tumor necrosis factor-α (TNF-α), and interferon-γ (IFN-γ)] within 24 hours of admission to ICU. The 28-day prognosis of the patients was followed up. The patients were divided into survival group and death group according to the prognosis. The clinical data between the two groups of sepsis patients with different prognosis were compared. Binary Logistic regression analysis was used to analyze the independent risk factors affecting the prognosis of patients with sepsis, and the receiver operator characteristic curve (ROC curve) was drawn to evaluate the predictive value of each risk factor for the prognosis of patients with sepsis. RESULTS: (1) A total of 227 patients with sepsis were enrolled, including 168 patients in the survival group (survival rate 74.0%) and 59 patients in the death group (mortality 26.0%). There were no significant differences in age (years old: 55.97±2.13 vs. 54.67±1.11) and gender (male: 71.2% vs. 57.1%) between the death group and the survival group (both P > 0.05), indicating that the baseline data of the two groups were comparable. (2) The APACHE II (19.37±0.99 vs. 14.88±0.61, P < 0.001) and PCT (µg/L: 12.39±2.94 vs. 4.14±0.90, P < 0.001) in the death group were significantly higher than those in the survival group, while the platelet count [PLT (×109/L): 144.75±12.50 vs. 215.99±11.26, P = 0.001] and thrombocytocrit [(0.14±0.01)% vs. (0.19±0.01)%, P = 0.001] were significantly lower than those in the survival group. (3) The level of IL-6 in the death group was significantly higher than that in the survival group (ng/L: 577.66±143.16 vs. 99.74±33.84, P < 0.001). There were no statistically significant differences in other cytokines, IL-2, IL-4, IL-10, TNF-α, IFN-γ and IL-17 between the death group and the survival group [IL-2 (ng/L): 2.44±0.38 vs. 2.63±0.27, P = 0.708; IL-4 (ng/L): 3.26±0.67 vs. 3.18±0.34, P = 0.913; IL-10 (ng/L): 33.22±5.13 vs. 39.43±2.85, P = 0.262; TNF-α (ng/L): 59.33±19.21 vs. 48.79±29.87, P = 0.839; IFN-γ (ng/L): 6.69±5.18 vs. 1.81±0.16, P = 0.100; IL-17 (ng/L): 2.05±0.29 vs. 2.58±0.33, P = 0.369]. (4) Binary Logistic regression analysis showed that APACHE II and IL-6 were independent risk factors affecting the prognosis of patients with sepsis [odds ratio (OR) and 95% confidence interval (95%CI) were 1.050 (1.008-1.093) and 1.001 (1.000-1.002), P values were 0.019 and 0.026, respectively]. (5) ROC curve analysis showed that APACHE II and IL-6 had certain predictive value for the prognosis of patients with sepsis, the area under the ROC curve (AUC) was 0.754 (95%CI was 0.681-0.827) and 0.592 (95%CI was 0.511-0.673), P values were < 0.001 and 0.035, respectively. When the optimal cut-off value of APACHE II was 16.50 score, the sensitivity was 72.6% and the specificity was 69.9%. When the optimal cut-off value of IL-6 was 27.87 ng/L, the sensitivity was 67.2% and the specificity was 52.8%. CONCLUSIONS: APACHE II score and IL-6 level have certain predictive value for the prognosis of patients with sepsis, the higher APACHE II score and IL-6 level, the greater the probability of death in patients with sepsis.

[Analysis of lymphocyte subsets in patients with sepsis and its impact on prognosis].

OBJECTIVE: To explore the characteristics of changes in peripheral blood lymphocyte subsets in patients with sepsis in intensive care unit (ICU) and analyze their predictive value for prognosis. METHODS: The clinical data of sepsis patients admitted to the surgical intensive care unit (SICU) of the First Affiliated Hospital of Zhengzhou University from January 2020 to December 2021 were analyzed retrospectively. The patients met the diagnostic criteria of Sepsis-3 and were ≥ 18 years old. Peripheral venous blood samples were collected from all patients on the next morning after admission to SICU for routine blood test and peripheral blood lymphocyte subsets. According to the 28-day survival, the patients were divided into two groups, and the differences in immune indexes between the two groups were compared. Logistic regression analysis was used to analyze the risk factors of immune indexes that affect prognosis. RESULTS: (1) A total of 279 patients with sepsis were enrolled in the experiment, of which 198 patients survived at 28 days (28-day survival rate 71.0%), and 81 patients died (28-day mortality 29.0%). There were no significant differences in age (years old: 57.81±1.71 vs. 54.99±1.05) and gender (male: 60.5% vs. 63.6%) between the death group and the survival group (both P > 0.05), and the baseline data was comparable.(2) Acute physiology and chronic health evalution II (APACHE II: 22.06±0.08 vs. 14.08±0.52, P < 0.001), neutrophil percentage [NEU%: (88.90±1.09)% vs. (84.12±0.77)%, P = 0.001], procalcitonin [PCT (µg/L): 11.97±2.73 vs. 5.76±1.08, P = 0.011], platelet distribution width (fL: 16.81±0.10 vs. 16.57±0.06, P = 0.029) were higher than those in the survival group, while lymphocyte percentage [LYM%: (6.98±0.78)% vs. (10.59±0.86)%, P = 0.012], lymphocyte count [LYM (×109/L): 0.70±0.06 vs. 0.98±0.49, P = 0.002], and platelet count [PLT (×109/L): 151.38±13.96 vs. 205.80±9.38, P = 0.002], and thrombocytocrit [(0.15±0.01)% vs. (0.19±0.07)%, P = 0.012] were lower than those in the survival group. (3) There was no statistically significant difference in the percentage of lymphocyte subsets between the death group and the survival group, but the absolute value of LYM (pieces/µL: 650.24±84.67 vs. 876.64±38.02, P = 0.005), CD3+ absolute value (pieces/µL: 445.30±57.33 vs. 606.84±29.25, P = 0.006), CD3+CD4+ absolute value (pieces/µL: 239.97±26.96 vs. 353.49±18.59, P = 0.001), CD19+ absolute value (pieces/µL: 111.10±18.66 vs. 150.30±10.15, P = 0.049) in the death group was lower than those in the survival group. Other lymphocyte subsets in the death group, such as CD3+CD8+ absolute value (pieces/µL: 172.40±24.34 vs. 211.22±11.95, P = 0.112), absolute value of natural killer cell [NK (pieces/µL): 101.26±18.15 vs. 114.72±7.64, P = 0.420], absolute value of natural killer T cell [NKT (pieces/µL): 33.22±5.13 vs. 39.43±2.85, P = 0.262], CD4-CD8- absolute value (pieces/µL: 41.07±11.07 vs. 48.84±3.31, P = 0.510), CD4+CD8+ absolute value (pieces/µL: 3.39±1.45 vs. 3.47±0.36, P = 0.943) were not significantly different from those in the survival group. (4)Logistic regression analysis showed that lymphocyte subsets were not selected as immune markers with statistical significance for the prognosis of sepsis. CONCLUSIONS: The changes of immune indexes in sepsis patients are closely related to their prognosis. Early monitoring of the above indexes can accurately evaluate the condition and prognosis of sepsis patients.

Diagnostic performance and clinical impact of blood metagenomic next-generation sequencing in ICU patients suspected monomicrobial and polymicrobial bloodstream infections.

Introduction: Early and effective application of antimicrobial medication has been evidenced to improve outcomes of patients with bloodstream infection (BSI). However, conventional microbiological tests (CMTs) have a number of limitations that hamper a rapid diagnosis. Methods: We retrospectively collected 162 cases suspected BSI from intensive care unit with blood metagenomics next-generation sequencing (mNGS) results, to comparatively evaluate the diagnostic performance and the clinical impact on antibiotics usage of mNGS. Results and discussion: Results showed that compared with blood culture, mNGS detected a greater number of pathogens, especially for Aspergillus spp, and yielded a significantly higher positive rate. With the final clinical diagnosis as the standard, the sensitivity of mNGS (excluding viruses) was 58.06%, significantly higher than that of blood culture (34.68%, P<0.001). Combing blood mNGS and culture results, the sensitivity improved to 72.58%. Forty-six patients had infected by mixed pathogens, among which Klebsiella pneumoniae and Acinetobacter baumannii contributed most. Compared to monomicrobial, cases with polymicrobial BSI exhibited dramatically higher level of SOFA, AST, hospitalized mortality and 90-day mortality (P<0.05). A total of 101 patients underwent antibiotics adjustment, among which 85 were adjusted according to microbiological results, including 45 cases based on the mNGS results (40 cases escalation and 5 cases de-escalation) and 32 cases on blood culture. Collectively, for patients suspected BSI in critical condition, mNGS results can provide valuable diagnostic information and contribute to the optimizing of antibiotic treatment. Combining conventional tests with mNGS may significantly improve the detection rate for pathogens and optimize antibiotic treatment in critically ill patients with BSI.

[Prognostic evaluation of coagulation indicators for patients with acute fatty liver of pregnancy].

OBJECTIVE: To explore the relevant clinical test indicators that affect the prognosis of patients with acute fatty liver of pregnancy (AFLP), and to provide a basis for early diagnosis and correct selection of treatment methods. METHODS: A retrospective analysis was conducted. Clinical data of AFLP patients in the intensive care unit (ICU) of the First Affiliated Hospital of Zhengzhou University from January 2010 to May 2021 were collected. According to the 28-day prognosis, the patients were divided into death group and survival group. The clinical data, laboratory examination indicators, and prognosis of the two groups were compared, and further binary Logistic regression analysis was used to analyze the risk factors affecting the prognosis of patients. At the same time, the values of related indicators at each time point (24, 48, 72 hours) after the start of treatment were recorded. The receiver operator characteristic curve (ROC curve) of prothrombin time (PT) and international normalized ratio (INR) for evaluating the prognosis of patients at each time point was drawn, and the area under the ROC curve (AUC) was calculated to evaluate the predictive value of relevant indicators at each time point for the prognosis of AFLP patients. RESULTS: A total of 64 AFLP patients were selected. The patients developed the AFLP during pregnancy (34.5±6.8) weeks, with 14 deaths (mortality of 21.9%) and 50 survivors (survival rate of 78.1%). There was no statistically significant difference in general clinical data between the two groups of patients, including age, time from onset to visit, time from visit to cessation of pregnancy, acute physiology and chronic health evaluations II (APACHE II), hospitalization time in ICU, and total hospitalization cost. However, the proportion of male fetuses and stillbirths in the death group was higher than that in the survival group. The laboratory examination indicators including the white blood cell count (WBC), alanine transaminase (ALT), serum creatinine (SCr), PT extension, INR elevation, and hyperammonia in the death group were significantly higher than those in the survival group (all P < 0.05). Through Logistic regression analysis of the above indicators showed that PT > 14 s and INR > 1.5 were risk factors affecting the prognosis of AFLP patients [PT > 14 s: odds ratio (OR) = 1.215, 95% confidence interval (95%CI) was 1.076-1.371, INR > 1.5: OR = 0.719, 95%CI was 0.624-0.829, both P < 0.01]. ROC curve analysis showed that both PT and INR at ICU admission and 24, 48, and 72 hours of treatment can evaluate the prognosis of AFLP patients [AUC and 95%CI of PT were 0.772 (0.599-0.945), 0.763 (0.608-0.918), 0.879 (0.795-0.963), and 0.957 (0.904-1.000), respectively; AUC and 95%CI of INR were 0.808 (0.650-0.966), 0.730 (0.564-0.896), 0.854 (0.761-0.947), and 0.952 (0.896-1.000), respectively; all P < 0.05], the AUC of PT and INR after 72 hours of treatment was the highest, with higher sensitivity (93.5%, 91.8%) and specificity (90.9%, 90.9%). CONCLUSIONS: AFLP often occurs in the middle and late stages of pregnancy, and the initial symptoms are mainly gastrointestinal symptoms. Once discovered, pregnancy should be terminated immediately. PT and INR are good indicators for evaluating AFLP patient efficacy and prognosis, and PT and INR are the best prognostic indicators after 72 hours of treatment.

Sinocyclocheiluslongicornus (Cypriniformes, Cyprinidae), a new species of microphthalmic hypogean fish from Guizhou, Southwest China.

Sinocyclocheiluslongicornussp. nov. is described from the Pearl River basin in Hongguo Town, Panzhou City, Guizhou Province, Southwest China. Based on the presence of the long horn-like structure on the back of the head, Sinocyclocheiluslongicornussp. nov. is assigned to the Sinocyclocheilusangularis species group. Sinocyclocheiluslongicornussp. nov. is distinguished from its congeners by a combination of morphological characters: (1) presence of a single, relatively long horn-like structure on the back of the head; (2) pigmentation absent; (3) reduced eyes; (4) dorsal-fin rays, ii, 7; (5) pectoral-fin rays, i, 13; (6) anal-fin rays, iii, 5; (7) pelvic-fin rays, i, 7; (8) lateral line pores 38-49; (9) gill rakers well developed, nine on first gill arch; and (10) tip of adpressed pelvic fin not reaching anus.

[Predictive value of the maximum aggregation rate of platelet for septic shock and septic shock with disseminated intravascular coagulation].

OBJECTIVE: To investigate the predictive value of the maximum aggregation rate (MAR) of platelet for septic shock and septic shock with disseminated intravascular coagulation (DIC). METHODS: A retrospective case-control study enrolled patients with sepsis admitted to department of critical care medicine of the First Affiliated Hospital of Zhengzhou University from January 2021 to November 2022. The basic data, dynamic platelet aggregation rate, blood routine, inflammation indicators, sequential organ failure assessment (SOFA) and other clinical indicators within 24 hours after admission were collected. Septic patients were divided into the shock group and the non-shock group according to the presence of septic shock; then refer to the International Society on Thrombosis and Hemostasis (ISTH) standard, patients with septic shock were divided into the shock DIC group and the shock non-DIC group according to the presence of dominant DIC. Compared the differences in platelet aggregation function between these groups, and the receiver operator characteristic curve (ROC curve) was drawn to evaluate the predictive value of the MAR for septic shock and septic shock with DIC. Spearman correlation analysis was used to analyze the correlation of MAR with inflammation indicators and the severity of illness in patients with sepsis. RESULTS: A total of 153 sepsis patients were included and 61 with septic shock (including 17 with dominant DIC and 44 without dominant DIC). Compared with the non-shock group, the level of procalcitonin (PCT), C-reactive protein (CRP), and SOFA score were significantly higher in the shock group [PCT (mg/L): 6.90 (2.50, 23.50) vs. 0.87 (0.26, 5.75), CRP (mg/L): 156.48 (67.11, 230.84) vs. 90.39 (46.43, 182.76), SOFA score: 11.00 (8.00, 14.00) vs. 5.00 (3.00, 8.00), all P < 0.05]. The platelet count (PLT) and the MAR induced by adenosine diphosphate (ADP), adrenaline (A), collagen (COL), and arachidonic acid (AA; ADP-MAR, A-MAR, COL-MAR, AA-MAR) in the shock group were significantly decreased [PLT (×109/L): 101.00 (49.00, 163.50) vs. 175.50 (108.25, 254.50), ADP-MAR: 28.50% (22.00%, 38.05%) vs. 45.90% (33.98%, 60.28%), A-MAR: 38.90% (30.00%, 55.40%) vs. 65.15% (54.38%, 72.53%), COL-MAR: 27.90% (20.85%, 36.55%) vs. 42.95% (33.73%, 54.08%), AA-MAR: 24.70% (16.40%, 34.20%) vs. 46.55% (28.33%, 59.20%), all P < 0.05]. Subgroup analysis revealed that, compared with the shock non-DIC group, the SOFA scores were significantly higher in patients in the shock DIC group (13.29±5.23 vs. 10.39±3.58, P < 0.05), the PLT and COL-MAR in the shock DIC group were significantly reduced [PLT (×109/L): 36.00 (22.00, 67.50) vs. 115.50 (84.25, 203.75), COL-MAR: 21.50% (17.85%, 32.60%) vs. 30.95% (22.98%, 38.53%), all P < 0.05]. ROC curve analysis showed that A-MAR had a higher predictive value for septic shock, and the area under the ROC curve (AUC) was 0.814 [95% confidence interval (95%CI) was 0.742-0.886, P = 0.000]. When the optimal cut-off value was 51.35%, the sensitivity was 68.9%, the specificity was 82.6%, the positive predictive value was 0.724 and the negative predictive value was 0.800. COL-MAR had some predictive value for septic shock with DIC, and the AUC was 0.668 (95%CI was 0.513-0.823, P = 0.044). When the optimal cut-off value was 21.90%, the sensitivity was 52.9%, the specificity was 79.5%, the positive predictive value was 0.500, and the negative predictive value was 0.813. Spearman correlation analysis showed that the MAR induced by each inducer was negatively correlated with inflammatory indicators and SOFA scores in sepsis patients, with A-MAR showing the strongest correlation with SOFA score (r = -0.327, P = 0.000). CONCLUSIONS: MAR, an indicator of platelet aggregation function, shows predictive value for septic shock and septic shock with DIC, and it could be used to for evaluating the severity of patients with sepsis. In addition, tt also can be used as a monitoring index to predict the changes of sepsis patients and to guide the treatment.

Molecular characterization and phylogenetic analysis of porcine epidemic diarrhea virus strains circulating in China from 2020 to 2021.

BACKGROUND: Porcine epidemic diarrhea virus (PEDV), an enteric coronavirus, has become the major causative agent of acute gastroenteritis in piglets since 2010 in China. RESULTS: In the current study, 91 complete spike (S) gene sequences were obtained from PEDV positive samples collected from 17 provinces in China from March 2020 to March 2021. A phylogenetic analysis showed that 92.3% (84 out of 91) of the identified strains belonged to GII subtype, while 7.7% (7 out of 91) were categorized as S-INDEL like strains and grouped within GI-c clade. Based on a recombination analysis, six of S-INDEL like strains were recombinant strains originated from S-INDEL strain FR/001/2014 and virulent strain AJ1102. In addition, PEDV variant strains (CH/GDMM/202012, CH/GXDX/202010 et al) carrying novel insertions (360QGRKS364 and 1278VDVF1281) in the S protein were observed. Furthermore, the deduced amino acid sequences for the S protein showed that multiple amino acid substitutions in the antigenic epitopes in comparison with the vaccine strains. CONCLUSIONS: In conclusion, these data provide novel molecular evidence on the epidemiology and molecular diversity of PEDV in 2020-2021. This information may help design a strategy for controlling and preventing the prevalence of PEDV variant strains in China.

Characterization and phylogenetic analysis of the complete mitochondrial genome of Sinocyclocheilus wenshanensis (Cypriniformes: Cyprinidae).

Sinocyclocheilus wenshanensis is a cyprinid fish species endemic to Southwestern China. In this study, we first sequenced and characterized the complete mitochondrial genome (mitogenome) of S. wenshanensis by next-generation sequencing method. The entire length of mitogenome is 16,595 base pairs (bp), containing 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and a control region. Its gene arrangement pattern was identical to other previously reported Sinocyclocheilus fishes. The overall base composition is 31.12% A, 16.63% G, 25.45% T, and 26.80% C, with AT content of 56.57%. Phylogenetic analysis using mitogenome of 26 Cyprinidae fishes showed that S. wenshanensis are closely related to S. aluensis and S. oxycephalus. This work would provide molecular information fundamental to future phylogenetic analyses among Sinocyclocheilus species.

Early Outcomes of Complex Vascular Reconstructions in Lower Extremities Using Spiral and Panel Vein Grafts.

BACKGROUND: Spiral saphenous vein grafts (SSVG) or paneled vein grafts (PVG) can be used when the diameter of the autologous great saphenous vein does not match the vessel that needs to be repaired. This study aimed to present early results of complex vascular reconstruction with SSVGs and PVGs in the lower extremities. METHODS: From May 2019 through January 2021, 6 SSVGs and 3 PVGs were used for vascular reconstruction in 9 patients. Patient data were collected retrospectively, including age, gender, cause of vascular pathology, target vessels, concomitant injury, surgical method, additional surgical methods, and hemodynamic status. The Kaplan-Meier method was used to calculate the rate of freedom from reintervention. RESULTS: Among these patients, 7 had trauma, 1 had graft infection, and 1 had vascular reconstruction after tumor excision. The mean duration of follow-up was 6 ± 6.6 months (range 1-19 months). The rate of freedom from reintervention for any reason was 77.8% at 1 year. Two patients underwent amputation after vascular reconstruction with patent vascular reconstructions. One of the 2 amputations was performed because of infection, and the other was due to ischemia >24 hr. The success rate of reconstruction was 100%, and the primary patency rate was 100%. The rate of limb salvage was 77.8%. There was no death, bleeding, embolism, skin ulcers, graft-related complication, or aneurysmal dilation during follow-up. CONCLUSIONS: SSVG and PVG were associated with low infection rates and satisfactory short-term patency rates. Both 2 grafts may be good choices when there is a diameter mismatch in vascular reconstructions.

A Next-Generation Sequencing of Plasma Exosome-Derived microRNAs and Target Gene Analysis with a Microarray Database of Thermally Injured Skins: Identification of Blood-to-Tissue Interactions at Early Burn Stage.

BACKGROUND: Plasma exosome-derived microRNA (miRNA) profiles following thermal injury and their relationship with gene expression derangements in burned skin remain unexplored. This study focused on the identification of key miRNA-mRNA axes in potential blood-to-tissue interactions at early burn stage. METHODS: Plasma exosomes were obtained from 6 severe burn patients 4-7 days post injury and 6 healthy volunteers. Next-generation sequencing (NGS) of exosomal small RNAs presented the differentially expressed miRNAs (DEMs). Target genes of the DEMs were predicted in the mirDIP database. Dataset GSE8056 was enrolled to acquire differentially expressed genes (DEGs) in burned skin compared to normal skin. Overlap between the DEGs and target genes of the DEMs were focus genes. The protein-protein interaction (PPI) network and enrichment analyses of the focus genes demonstrated hub genes and suggested underlying mechanisms and pathways. The hub genes and upstream DEMs were selected to construct key miRNA-mRNA axes. RESULTS: The NGS of plasma exosome-derived small RNAs identified 85 DEMs (14 downregulated miRNAs and 71 upregulated miRNAs) with 12,901 predicted target genes. Dataset GSE8056 exhibited 1861 DEGs in partial-thickness burned skins 4-7 days postburn. The overlap between DEGs and target genes of DEMs displayed 1058 focus genes. The top 9 hub genes (CDK1, CCNB1, CCNA2, BUB1B, PLK1, KIF11, AURKA, NUSAP1 and CDCA8) in the PPI network of the focus genes pointed to 16 upstream miRNAs in DEMs, including 4 downregulated miRNAs (hsa-miR-6848-3p, has-miR-4684-3p, has-miR-4786-5p and has-miR-365a-5p) and 12 upregulated miRNAs (hsa-miR-6751-3p, hsa-miR-718, hsa-miR-4754, hsa-miR-6754-3p, hsa-miR-4739, hsa-miR-6739-5p, hsa-miR-6884-3p, hsa-miR-1224-3p, hsa-miR-6878-3p, hsa-miR-6795-3p, hsa-miR-550a-3p, and hsa-miR-550b-3p). A key miRNA-mRNA network in potential blood-to-tissue interactions at early burn stage was therefore constructed. CONCLUSION: An NGS and bioinformatic analysis in the study identified key miRNA-mRNA axes in potential blood-to-tissue interactions at early burn stage, suggesting plasma exosome-derived miRNAs may impact on the alteration patterns of gene expressions in a burn wound.

Beneficial Effect of Immune-Enhanced Enteral Nutrition on Immune Function in Patients With Severe Neurological Diseases: A Single-Center Randomized Controlled Trial.

Background: Undernutrition is the main reason for the use of artificial nutrition in patients with severe neurological diseases. However, the clinical and immunological outcomes of enteral nutrition supplemented with immunomodulatory nutrients in these patients remain unclear. Methods: In this single-center, randomized controlled study, 57 patients with severe neurological diseases were randomly divided into the following two groups according to the type of enteral nutrition they would receive: immune-enhancing (IE) (n = 27) and non-IE (NIE) (n = 30). The IE and NIE groups received enteral nutrition supplemented with immunomodulatory nutrients and standard enteral nutrition, respectively. We compared the nutritional status and the state of cellular immunity between the patients of the two groups. Clinical and immunological variables were evaluated following nutritional treatment. Results: Feeding intolerance was lower in the IE than that in the NIE group (P = 0.04). However, there were no significant differences between the results of the two groups in terms of length of stay in the intensive care unit or hospital, extubation time, or body mass index (P > 0.05). The CD4+ T-lymphocyte count and CD4+/CD8+ ratio in the peripheral blood increased significantly in the IE group. The expression of CD28 activated cell surface markers was higher in the IE than in the NIE group. In addition, increased plasma interferon-γ levels were recorded in the IE group, whereas the levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-8, and IL-10 decreased. Conclusions: Immune-enhanced enteral nutrition could improve the immune status and feeding tolerance in patients with severe neurological diseases. Trial Registration:www.chictr.org.cn/index.aspx, identifier: ChiCTR-IPR-17013909.

Trehalose-6-phosphate phosphatase inhibitor: N-(phenylthio) phthalimide, which can inhibit the DON biosynthesis of Fusarium graminearum.

Fusarium head blight(FHB)caused by Fusarium graminearum species complex (FGSC) is one of the most important diseases around the world. Deoxynivalenol (DON) is a type of mycotoxin produced by FGSC when infecting cereal crops. It is a serious threat to the health of both humans and livestock. Trehalose-6-phosphate phosphatase (TPP), a conserved metabolic enzyme found in many plants and pathogens, catalyzes the formation of trehalose. N-(phenylthio) phthalimide (NPP) has been reported to inhibit the normal growth of nematodes by inhibiting the activity of TPP, but this inhibitor of nematodes has not previously been tested against F. graminearum. In this study, we found that TPP in F. graminearum (FgTPP) had similar secondary structures and conserved cysteine (Cys356) to nematodes by means of bioinformatics. At the same time, the sensitivity of F. graminearum strains to NPP was determined. NPP exhibited a better inhibitory effect on conidia germination than mycelial growth. In addition, the effects of NPP on DON biosynthesis and trehalose biosynthesis pathway in PH-1 were also determined. We found that NPP decreased DON production, trehalose content, glucose content and TPP enzyme activity but increased trehalose-6-phosphate content and trehalose-6-phosphate synthase (TPS) enzyme activity. Moreover, the expression of TRI1, TRI4, TRI5, TRI6, and TPP genes were downregulated, on the contrary, the TPS gene was upregulated. Finally, in order to further determine the control ability of NPP on DON production in the field, we conducted a series of field experiments, and found that NPP could effectively reduce the DON content in wheat grain and had a general control effect on FHB. In conclusion, the research in this study will provide important theoretical basis for controlling FHB caused by F. graminearum and reducing DON production in the field.

Case Report: A Severe and Multi-Site Nocardia farcinica Infection Rapidly and Precisely Identified by Metagenomic Next-Generation Sequencing.

Nocardia genus is an aerobic, gram-positive, and opportunistic pathogen, which mainly affects cell-mediated immunosuppressed patients. Early diagnosis and treatment greatly improve prognosis. However, the limitation of golden standard-bacterial culture exists. Here, we report a 61-year-old male with pneumonia, sepsis and intermuscular abscesses induced by Nocardia farcinica. Venous blood culture reported negative results. Former improper diagnosis and treatment did not improve his condition. With the assistant of metagenomic next-generation sequencing, the pathogen was identified as Nocardia farcinica. He was then applied with accurate treatment and had a remarkable clinical and radiological improvement.

[Risk factors for death and their predictive value on diabetic kidney disease patients in intensive care unit based on MIMIC-III database].

OBJECTIVE: To analyze the influencing factors of prognosis of patients with diabetic kidney disease (DKD) in intensive care unit (ICU), and analyze their predictive value. METHODS: Based on the inpatient information of more than 50 000 patients from June 2001 to October 2012 in the latest version of American Intensive Care Medical Information Database (MIMIC-III v1.4), the data of DKD patients were screened out, including gender, age, body weight, comorbidities [hypertension, coronary heart disease, chronic obstructive pulmonary disease (COPD), chronic kidney disease (CKD)], sequential organ failure assessment (SOFA) score, the length of ICU stay, the incidence of mechanical ventilation, vasoactive drugs and renal replacement therapy during the ICU hospitalization, complications of other diseases [ventilator-associated pneumonia (VAP), urinary tract infection (UTI), diabetic ketoacidosis (DKA), acute myocardial infarction (AKI)] and prognosis of ICU. At the same time, the blood routine and biochemical data of the first 24 hours in ICU and the extremum values during the ICU hospitalization were collected. Multivariate Logistic regression analysis was used to screen the prognostic factors of DKD patients in ICU, and receiver operating characteristic (ROC) curve was plotted to analyze the predictive value of death risk factors. RESULTS: 416 DKD patients were screened out, 20 patients were excluded due to data missing, and finally 396 patients were enrolled, including 220 survival patients and 176 dead patients. Compared with the survival group, the patients in the death group were older (years old: 57.13±13.04 vs. 52.61±14.15), with lower rates of hypertension and CKD (11.4% vs. 23.6%, 26.7% vs. 41.4%), higher SOFA scores and baseline values of blood urea nitrogen (BUN), serum creatinine (SCr) and blood K+ [SOFA score: 5.86±2.79 vs. 4.49±2.56, BUN (mmol/L): 18.4±10.0 vs. 14.8±9.0, SCr (µmol/L): 387.2±382.8 vs. 284.6±244.9, K+ (mmol/L): 4.64±0.99 vs. 4.33±0.86], and longer ICU stay [days: 2.65 (1.48, 5.21) vs. 2.00 (1.00, 4.00)], and the differences were statistically significant (all P < 0.01). Further analysis of laboratory tests extremum values during ICU hospitalization showed that the maximum (max) and minimum (min) values of white blood cell (WBC), BUN and SCr, and K+max in the death group were significantly higher than those in the survival group [WBCmax (×109/L): 17.3±10.3 vs. 14.5±7.3, WBCmin (×109/L): 7.9±4.1 vs. 6.7±2.7, BUNmax (mmol/L): 23.8±10.4 vs. 18.8±10.2, BUNmin (mmol/L): 11.0±6.6 vs. 9.3±6.6, SCrmax (µmol/L): 459.7±392.5 vs. 350.1±294.4, SCrmin (µmol/L): 246.6±180.3 vs. 206.9±195.4, K+max (mmol/L): 5.35±0.93 vs. 5.09±0.99], and the minimum values of hemoglobin (Hbmin) and glucose (Glumin) were significantly lower than those in the survival group [Hbmin (g/L): 87.4±14.5 vs. 90.6±16.5, Glumin (mmol/L): 4.0±1.7 vs. 4.6±2.0], and the differences were statistically significant (all P < 0.05). The incidences of mechanical ventilation and vasoactive drugs during ICU hospitalization in the death group were significantly higher than those in the survival group (37.5% vs. 24.1%, 32.4% vs. 20.0%, both P < 0.01), and the incidences of UTI and AMI in the death group were significantly higher than those in the survival group (29.5% vs. 19.1%, 8.5% vs. 3.6%, both P < 0.05). Multivariate Logistic regression analysis showed that age [odds ratio (OR) = 1.019, 95% confidence interval (95%CI) was 1.003-1.036, P = 0.023], SOFA score (OR = 1.142, 95%CI was 1.105-1.246, P = 0.003), WBCmin (OR = 1.134, 95%CI was 1.054-1.221, P = 0.001) and BUNmax (OR = 1.010, 95%CI was 1.002-1.018, P = 0.018) were risk factors of death of DKD patients in ICU. ROC curve analysis showed that the area under ROC curve (AUC) of combination of risks factors of death was 0.706, the sensitivity was 61.6%, and the specificity was 73.2%. CONCLUSIONS: In order to prevent DKD patients from getting worse in ICU, we should pay close attention to the blood biochemical indexes, especially the renal function indexes, and give timely treatment. At the same time, we should actively prevent the occurrence of complications such as infection and cardiovascular disease.

[Effect of Xuebijing on inflammatory response and prognosis in patients with septic shock].

OBJECTIVE: To study the effect of Xuebijing on inflammatory response and prognosis in patients with septic shock. METHODS: A prospective randomized controlled study was conducted. Eighty septic shock patients admitted to department of critical care medicine of the First Affiliated Hospital of Zhengzhou University from January to December in 2019 were enrolled. The enrolled patients were divided into Xuebijing group and control group by randomized number table method, with 40 cases in each group. Both groups were strictly followed the guidelines for the diagnosis and treatment of septic shock to take comprehensive treatment measures against sepsis. On this basis, Xuebijing group received intravenous 100 mL Xuebijing injection twice a day for 7 days. Baseline data of enrolled patients were recorded. The levels of interleukin-6 (IL-6), procalcitonin (PCT), C-reactive protein (CRP) and heparin binding protein (HBP) were measured before treatment and 3, 7 and 10 days after treatment. Mechanical ventilation time, the length of intensive care unit (ICU) stay, total hospitalization time and 28-day mortality were recorded. The differences of every indicator between the two groups were compared. Independent risk factors affecting patient prognosis were analyzed by binary Logistic regression. RESULTS: (1) There was no significant difference in baseline data such as gender, age, infection site, acute physiology and chronic health evaluation II (APACHE II) and sequential organ failure score (SOFA) between the two groups. (2) The levels of serum inflammatory factors in both groups showed a decreasing trend after treatment. Compared with the control group, IL-6 and HBP in the Xuebijing group significantly decreased on day 7 [IL-6 (ng/L): 66.20 (16.34, 163.71) vs. 79.81 (23.95, 178.64), HBP (ng/L): 95.59 (45.23, 157.37) vs. 132.98 (73.90, 162.05), both P < 0.05]; on day 10, PCT, CRP, IL-6 and HBP significantly decreased [PCT (µg/L): 1.14 (0.20, 3.39) vs. 1.31 (0.68, 4.21), CRP (mg/L): 66.32 (19.46, 115.81) vs. 89.16 (20.52, 143.76), IL-6 (ng/L): 31.90 (13.23, 138.74) vs. 166.30 (42.75, 288.10), HBP (ng/L): 62.45 (29.17, 96.51) vs. 112.33 (58.70, 143.96), all P < 0.05]. (3) Compared with the control group, mechanical ventilation time and the length of ICU stay were significantly shortened and the total hospitalization expenses were significantly reduced in Xuebijing group [mechanical ventilation time (hours): 57.0 (0, 163.5) vs. 168.0 (24.0, 282.0), the length of ICU stay (days): 8.80±4.15 vs. 17.13±7.05, the total hospitalization expenses (ten thousand yuan): 14.55±7.31 vs. 20.01±9.86, all P < 0.05]. There was no significant difference in 28-day mortality and the total hospitalization time [28-day mortality: 37.5% vs. 35.0%, the total hospitalization time (days): 13.05±8.44 vs. 18.30±9.59, both P > 0.05]. (4) Patients were divided into death and survival groups according to the prognosis, and univariate analysis showed that white blood cell (WBC), neutrophil percentage (NEU%), CRP, lactic acid (Lac), APACHE II score, IL-6, HBP were the factors influencing the prognosis of patients. The above indicators were further analyzed by Logistic regression, which showed that CRP, IL-6, and APACHE II score were independent risk factors for prognosis [odds ratio (OR) was 1.007, 1.828, 1.229, all P < 0.05]. CONCLUSIONS: Combined with Xuebijing to treat septic shock can reduce the body's inflammatory response to a certain extent, thereby reducing the time of mechanical ventilation, shortening the stay of ICU and reducing the total cost of hospitalization. But it cannot reduce the 28-day mortality of patients with septic shock.

[The role of recombinant human thrombopoietin in critically ill patients with sepsis-associated thrombocytopenia: a clinical study].

OBJECTIVE: To investigate the effect of recombinant human thrombogenin (rhTPO) on sepsis-associated thrombocytopenia. METHODS: A prospective randomized controlled study was conducted. One hundred patients with sepsis-associated thrombocytopenia admitted to the department of critical care medicine of the First Affiliated Hospital of Zhengzhou University from August 2019 to October 2020 were enrolled. The enrolled patients were divided into rhTPO-using group (TPO group) and routine group (control group) by random number table method, with 50 cases in each group. Both groups were treated according to the guideline of Sepsis-3. In addition, TPO group received rhTPO 15 000 U, once daily for 7 days. Geneal information and acute physiology and chronic health evaluation II (APACHE II) were recorded. The levels of platelet count (PLT), blood coagulation function [prothrombin time (PT) and prothrombin activity (PTA)], myocardial enzyme indexes [troponin (Tn) and creatine kinase (CK)], liver and kidney function [aspartate aminotransferase (AST), total bilirubin (TBil) and creatinine (Cr)] and inflammatory biomarkers [procalcitonin (PCT) and C-reactive protein (CRP)] were recorded before treatment and 1, 3, 5 and 7 days after treatment. The infusion volume of blood components, duration of mechanical ventilation, length of stay in ICU, total length of hospitalization, total cost of hospitalization and 28-day outcome were recorded. According to whether the PLT was lower than 50×109/L, the patients in TPO group were divided into the TPO A group (PLT ≥ 50×109/L, 16 cases) and TPO B group (PLT < 50×109/L, 34 cases), and the absolute value of PLT increase, duration of mechanical ventilation, length of stay in ICU, total length of hospitalization, total cost of hospitalization and 28-day outcome of the two groups were compared. RESULTS: (1) In TPO and control groups, there were no statistically significant differences in gender, age, proportion of patients with primary infection site, APACHEII score, PLT, coagulation function, myocardial enzymes, liver and kidney function and inflammation indexes before treatment (all P > 0.05). (2) The PLT levels of the TPO group were significantly higher than those of the control group on the 5th and 7th day after treatment (×109/L: day 5, 63.94±44.01 vs. 49.85±29.26, day 7, 125.85±112.31 vs. 76.81±50.87, both P < 0.05), and there were no statistically significant differences in PT, PTA, Tn, CK, AST, TBil, Cr, PCT or CRP before and on the 1, 3, 5, 7 days after treatment between TPO and control groups (all P > 0.05). (3) The amount of platelet transfusion in the TPO group was lower than that in the control group [treatment amount: 0 (0, 0) vs 0 (0, 2.00), P = 0.001]. (4) There were no statistically significant differences in mechanical ventilation time, length of stay in ICU, total length of hospitalization, total cost of hospitalization or 28-day outcome between TPO and control groups (all P > 0.05). The mechanical ventilation time, ICU stay time and total hospitalization time of TPO A group were longer than those in TPO B group, but the differences were not statistically significant [mechanical ventilation time (hours): 131.00 (0, 311.00) vs. 50.00 (0, 192.00), ICU stay time (days): 14.44±8.57 vs. 11.73±9.24, total hospitalization time (days): 15.00 (6.00, 23.50) vs. 18.00 (8.00, 31.00), all P > 0.05]. The absolute value of PLT increase in TPO A group was higher than that of TPO B group, but the difference was not statistically significant [×109/L: 65.00 (16.50, 131.50) vs. 36.00 (18.00, 130.00), P > 0.05]. CONCLUSIONS: RhTPO can significantly increase the PLT of patients with sepsis-related thrombocytopenia, thereby reduce the amount of platelet transfusion, but it cannot shorten the length of ICU stay time and total hospitalization time, and it cannot reduce 28-day mortality.

Pareuchiloglanis (Teleostei: Sisoridae) from the Pearl River, China with description of three new species.

We describe three new species of Pareuchiloglanis. Based on a comparison of 17 valid species of Pareuchiloglanis, the genus can be divided into two groups contingent on their gill opening size and the anus position. One group, which we call the large gill opening group, has a large gill opening extending to the base of the first pectoral-fin element; the anus is obviously closer to pelvic-fin insertion than the anal-fin origin; this group includes five species distributed in the Red and Pearl Rivers, China. The other group has a small gill opening extending only to the middle base of the pectoral-fin elements; the anus is usually located at the midpoint of the pelvic-fin insertion to the anal-fin origin or slightly behind. This group includes the other 12 species, which are distributed in the Mekong and Yangtze Rivers. The large-gill-opening group can be divided into two sub-groups based on the length of the caudal peduncle. One sub-group has a long caudal-peduncle and the distance from the anal-fin origin to caudal-fin base is greater than distance from the pelvic-fin insertion. This sub-group is only distributed in the Pearl River drainage. Another sub-group has a short caudal peduncle and the distance from the anal-fin origin to the caudal-fin base is typically smaller than the distance from the pelvic-fin insertion. This sub-group is only distributed in the Red River basin of China and Vietnam. The former will be called the large-gill-opening group with long caudal peduncle in the text and only includes one species P. longicauda. During our ongoing taxonomic work of specimens collected from Nanpan-jiang and Beipan-jiang (upper Pearl River drainage in Yunnan, China), some Pareuchiloglanis specimens that had the characters of the large-gill-opening group with long caudal peduncle represent three undescribed species.

Testing the validity of two putative sympatric species from Sinocyclocheilus (Cypriniformes: Cyprinidae) based on mitochondrial cytochrome b sequences.

There are over 60 species within the freshwater fish genus Sinocyclocheilus (Cypriniformes: Cyprinidae) distributed throughout the Yunnan-Guizhou Plateau and its surrounding areas in China. In recent years, the increasing number of new species described has raised some controversy about the validity of several species within this genus, notably the putative sympatric species pair S. qiubeiensis and S. jiuchengensis. To test the validity of S. qiubeiensis and S. jiuchengensis, we analyzed the complete sequences of the mitochondrial cytochrome b (CYTB) gene of 20 identified species and one outgroup species. Phylogenetic relationships were reconstructed using CYTB with maximum likelihood (ML) and Bayesian inference (BI) methods. Our phylogenetic results showed that all individuals of S. qiubeiensis and S. jiuchengensis clustered in one clade with strong support. In addition, the genetic distance between the two species was 0.11%, within the range observed at the intraspecific level. The most recent common ancestor of S. qiubeiensis and S. jiuchengensis dated back to 0.13 million years ago, indicating little time for complete speciation to have occurred. These results clearly support the viewpoint that S. jiuchengensis is a synonym of S. qiubeiensis.

[Effect of pyrrolidine dithiocarbamate on oxidative stress and mitochondrial function of lung tissue in mice with acute lung injury].

OBJECTIVE: To investigate the effects of pyrrolidine dithiocarbamate (PDTC) on oxidative stress and mitochondrial function of lung tissue in mice with acute lung injury (ALI) induced by lipopolysaccharide (LPS). METHODS: Forty female Balb/c mice were randomly divided into normal saline (NS) control group, LPS model group, PDTC group, and PDTC+LPS group, with 10 mice in each group. The model of mice with ALI was reproduced by intraperitoneal injection of 15 mg/kg LPS. PDTC was administered intraperitoneally with 50 mg/kg PDTC 1 hour before LPS treatment in the PDTC+LPS group. The mice in NS control group was given intraperitoneal injection of 0.1 mL NS only, and those in PDTC group was given intraperitoneal injection of 50 mg/kg PDTC only. The mice were sacrificed at 24 hours after model reproduction, and the lung tissues were harvested. The total antioxidant capacity (T-AOC) of lung tissue was measured by spectrophotometric kits. The content of malondialdehyde (MDA) was determined by thiobarbituric acid reactive substances assay. The protein expressions of superoxide dismutases (SOD1, SOD2) and catalase (CAT) in lung tissue were determined by Western Blot. Mitochondria from mouse lungs were isolated, and adenosine triphosphate (ATP) synthesis was measured with a luciferase/luciferin-based approach. The mitochondrial membrane potential (ΔΨm) was estimated by using Rhodamine. The mRNA expressions of mitochondrial uncoupling proteins (UCPs) were determined by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: LPS stimulation could significantly increase oxidative stress in lung tissue of mice and lead to mitochondrial dysfunction. The results showed that the protein expressions of T-AOC and SOD1 were decreased, the level of MDA was increased, the ATP synthesis was decreased in the mitochondrial, the ΔΨm was decreased, and the mRNA expression of UCP2 was decreased. However, there was no significant change in the expressions of SOD2, CAT in lung tissue and UCP1, UCP3 in the mitochondria. Pretreatment with PDTC could obviously alleviate the increase in LPS-induced oxidative stress in lung tissue and mitigate mitochondrial dysfunction. Compared with the LPS model group, T-AOC in lung tissue of PDTC+LPS group was significantly increased (U/g: 0.35±0.08 vs. 0.31±0.07), the level of MDA was significantly decreased (µmol/mg: 13.29±1.13 vs. 17.54±1.72), the protein expression of SOD1 was significantly upregulated (SOD1 protein: 1.13±0.11 vs. 0.71±0.09), ATP synthesis was significantly increased in the mitochondrial (µmol/mg: 49.23±5.42 vs. 36.92±2.21), ΔΨm was significantly increased (mV: 226.03±11.69 vs. 194.86±7.79), and the mRNA expression of UCP2 was significantly increased (2-ΔΔCt: 0.88±0.06 vs. 0.73±0.04). The differences were statistically significant (all P < 0.05). In lung tissue of normal mice, PDTC treatment also had the effect of anti-oxidizing, reducing oxidative stress and promoting ATP synthesis in the mitochondrial. Compared with the NS control group, the level of T-AOC (U/g: 0.49±0.09 vs. 0.43±0.06) and the protein expressions of SOD2 and CAT (SOD2 protein: 1.33±0.08 vs. 1.00±0.11, CAT protein: 1.39±0.08 vs. 1.00±0.11), and ATP synthesis in the mitochondrial of PDTC group was significantly increased (µmol/mg: 61.53±4.92 vs. 53.33±3.20), MDA was significantly decreased (µmol/mg: 10.27±1.25 vs. 12.27±1.36), with statistical differences, but had no effect on the protein expression of SOD1 in lung tissue and ΔΨm and UCPs mRNA expressions in mitochondrion. CONCLUSIONS: LPS can induce ALI in mice, increased oxidative stress in lung tissue, and induce mitochondrial dysfunction by inhibiting ATP synthesis. PDTC pretreatment has anti-oxidative effect on LPS-induced ALI, and can mitigate mitochondrial dysfunction.