Novel Gene Clusters for Natural Product Synthesis Are Abundant in the Mangrove Swamp Microbiome.

Natural microbial communities produce a diverse array of secondary metabolites with ecologically and biotechnologically relevant activities. Some of them have been used clinically as drugs, and their production pathways have been identified in a few culturable microorganisms. However, since the vast majority of microorganisms in nature have not been cultured, identifying the synthetic pathways of these metabolites and tracking their hosts remain a challenge. The microbial biosynthetic potential of mangrove swamps remains largely unknown. Here, we examined the diversity and novelty of biosynthetic gene clusters in dominant microbial populations in mangrove wetlands by mining 809 newly reconstructed draft genomes and probing the activities and products of these clusters by using metatranscriptomic and metabolomic techniques. A total of 3,740 biosynthetic gene clusters were identified from these genomes, including 1,065 polyketide and nonribosomal peptide gene clusters, 86% of which showed no similarity to known clusters in the Minimum Information about a Biosynthetic Gene Cluster (MIBiG) repository. Of these gene clusters, 59% were harbored by new species or lineages of Desulfobacterota-related phyla and Chloroflexota, whose members are highly abundant in mangrove wetlands and for which few synthetic natural products have been reported. Metatranscriptomics revealed that most of the identified gene clusters were active in field and microcosm samples. Untargeted metabolomics was also used to identify metabolites from the sediment enrichments, and 98% of the mass spectra generated were unrecognizable, further supporting the novelty of these biosynthetic gene clusters. Our study taps into a corner of the microbial metabolite reservoir in mangrove swamps, providing clues for the discovery of new compounds with valuable activities. IMPORTANCE At present, the majority of known clinical drugs originated from cultivated species of a few bacterial lineages. It is vital for the development of new pharmaceuticals to explore the biosynthetic potential of naturally uncultivable microorganisms using new techniques. Based on the large numbers of genomes reconstructed from mangrove wetlands, we identified abundant and diverse biosynthetic gene clusters in previously unsuspected phylogenetic groups. These gene clusters exhibited a variety of organizational architectures, especially for nonribosomal peptide synthetase (NRPS) and polyketide synthase (PKS), implying the presence of new compounds with valuable activities in the mangrove swamp microbiome.

Prevention and control of COVID-19 by primary health care facilities in China: a field-survey-based qualitative study in three typical cities.

BACKGROUND: During the coronavirus disease 2019 (COVID-19) containment, primary health care (PHC) facilities inChina played an important role in providing both healthcare and public care services to community populations. The tasks of COVID-19 containment facilitated by PHC facilities were different among different regions and during different periods of COVID-19 pandemic. We sought to investigate the gaps on task participation, explore existing problems and provide corresponding solutions. METHODS: Semi-structured face-to-face interviews with COVID-19 prevention and control management teams of PHC facilities were conducted. Purposive stratified sampling was used and 32 team members of 22 PHC facilities were selected from Wuhan (as high-risk city), Shanghai (as medium-risk city) and Zunyi (as low-risk city). Framework analysis was employed to analyze the transcribed recordings. RESULTS: The main tasks of PHC facilities during the early period of the pandemic included assisting in contact tracing and epidemiological investigation, screening of populations at high-risk at travel centers/internals, house-by-house, or pre-examination/triage within PHC facilities; at-home/ centralized quarantine management; the work of fever sentinel clinics. Further analyses revealed the existing problems and suggestions for improvement or resolutions. Regular medical supply reserves were recommended because of the medical supply shortage during the pre-outbreak period. Temporarily converted quarantine wards and centralized quarantine centers could be used to deal with pressures on patients' treatment and management of the febrile patients. Only after strict evaluation of nucleic acid testing (NAT) results and housing conditions, decision on quarantine at-home or centralized quarantine centers could be made. Settings of fever sentinel clinics at PHC facilities allowed fever patients with no COVID-19 infection risks for treatment without being transferred to fever clinics of the designed secondary hospitals. Psychological intervention was sometimes in need and really helped in addressing individuals' mental pressures. CONCLUSIONS: During the COVID-19 containment, PHC facilities in China were responsible for different tasks and several problems were encountered in the working process. Accordingly, specific and feasible suggestions were put forward for different problems. Our findings are highly beneficial for healthcare teams and governments in handling similar situations.

Preoperative prediction of tumour deposits in rectal cancer by an artificial neural network-based US radiomics model.

OBJECTIVE: To develop a machine learning-based ultrasound (US) radiomics model for predicting tumour deposits (TDs) preoperatively. METHODS: From December 2015 to December 2017, 127 patients with rectal cancer were prospectively enrolled and divided into training and validation sets. Endorectal ultrasound (ERUS) and shear-wave elastography (SWE) examinations were conducted for each patient. A total of 4176 US radiomics features were extracted for each patient. After the reduction and selection of US radiomics features , a predictive model using an artificial neural network (ANN) was constructed in the training set. Furthermore, two models (one incorporating clinical information and one based on MRI radiomics) were developed. These models were validated by assessing their diagnostic performance and comparing the areas under the curve (AUCs) in the validation set. RESULTS: The training and validation sets included 29 (33.3%) and 11 (27.5%) patients with TDs, respectively. A US radiomics ANN model was constructed. The model for predicting TDs showed an accuracy of 75.0% in the validation cohort. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and AUC were 72.7%, 75.9%, 53.3%, 88.0% and 0.743, respectively. For the model incorporating clinical information, the AUC improved to 0.795. Although the AUC of the US radiomics model was improved compared with that of the MRI radiomics model (0.916 vs. 0.872) in the 90 patients with both ultrasound and MRI data (which included both the training and validation sets), the difference was nonsignificant (p = 0.384). CONCLUSIONS: US radiomics may be a potential model to accurately predict TDs before therapy. KEY POINTS: • We prospectively developed an artificial neural network model for predicting tumour deposits based on US radiomics that had an accuracy of 75.0%. • The area under the curve of the US radiomics model was improved than that of the MRI radiomics model (0.916 vs. 0.872), but the difference was not significant (p = 0.384). • The US radiomics-based model may potentially predict TDs accurately before therapy, but this model needs further validation with larger samples.

Malignancy risk stratification and FNA recommendations for thyroid nodules: A comparison of ACR TI-RADS, AACE/ACE/AME and ATA guidelines.

OBJECTIVE: To compare diagnostic performance and malignancy risk stratification among guidelines set forth by the American Thyroid Association (ATA) in 2015, the American Association of Clinical Endocrinologists (AACE), the American College of Endocrinology (ACE) and the Association Medici Endocrinologi (AME) in 2016, and the American College of Radiology (ACR) in 2017. METHODS: The retrospective study was approved by the hospital ethics committee, and the informed consent requirement was waived. From October 2015 to March 2016, a total of 230 patients with 230 consecutive thyroid nodules were enrolled in this study. Each nodule was classified by one junior and one senior radiologist separately according to ACR TI-RADS, AACE/ACE/AME and ATA guidelines. The malignancy diagnostic performance and the number of FNA recommendations were pairwise compared among three guidelines using chi-square tests. RESULTS: Of the 230 thyroid nodules, 137 were malignant, and 93 were benign. However, 19.6% of the nodules (45 of 230) did not match any pattern using the ATA guidelines but with a high risk of malignancy (68.9%). The ACR TI-RADS derived the highest diagnostic performance, from both junior radiologist (AUC 0.815) and senior radiologist (AUC 0.864). The ACR guidelines also showed the greatest level of sensitivity (junior: 86.1%, senior: 94.9%), compared with AACE/ACE/AME and ATA guidelines. The number of thyroid nodules recommended to fine-needle aspiration (FNA) was the lowest (37.8%, 40.4%) by ACR TI-RADS, and meanwhile, the malignant detection rate within these nodules was highest (64.4%, 68.8%). CONCLUSIONS: The ACR guidelines present a higher level of diagnostic indicators and may offer a meaningful reduction in FNA recommendations with a higher malignancy detection rate.

Carotid Artery Stiffness Assessment by Ultrafast Ultrasound Imaging: Feasibility and Potential Influencing Factors.

OBJECTIVES: To evaluate the feasibility of the ultrafast ultrasound pulsed wave velocity (PWV) for carotid stiffness assessment and potential influencing factors. METHODS: Ultrafast PWV measurements of 442 carotid arteries in 162 consecutive patients (patient group) and 66 healthy volunteers (control group) were performed. High- and very high-frequency transducers were used in 110 carotid segments. The ultrafast PWVs at the beginning and end of systole were automatically measured. The correlations between the intima-media thickness (IMT) and ultrafast PWV and the equipment and carotid factors influencing the utility of ultrafast PWV were analyzed. RESULTS: Each ultrafast PWV acquisition was completed within 1 minute. The intraobserver variability showed mean differences ± SD of 0.12 ± 1.28 m/s for the PWV before systole and 0.06 ± 1.30 m/s for the PWV at the end of systole. Ultrafast PWV measurements were more likely obtained with the very high- frequency transducer when the IMT was less than 1.5 mm (P < .05). A generalized linear mixed-effects model analysis showed that the very high-frequency transducer had a greater ability to obtain a valid carotid ultrafast PWV measurement with an IMT of less than 1.5 mm (P < .05). The IMT was positively correlated with the PWV before systole and at the end of systole (r = 0.207-0.771; all P < .05) in the control group, patient group, and carotid subgroup with an IMT of less than 1.5 mm. A multiple regression analysis showed that the IMT and plaque were important independent factors in predicting failure of the ultrafast PWV (P < .001). CONCLUSIONS: The ultrafast PWV is an effective and user-friendly method for evaluating carotid stiffness. The IMT and transducer type are factors influencing the ability to obtain an ultrafast PWV measurement.

Optimizing the US Diagnosis of Biliary Atresia with a Modified Triangular Cord Thickness and Gallbladder Classification.

Purpose To evaluate the diagnostic performance of ultrasonography (US) in the identification and exclusion of biliary atresia with a modified triangular cord thickness metric together with a gallbladder classification scheme, as well as hepatic artery (HA) diameter and liver and spleen size, in a large sample of jaundiced infants. Materials and Methods The ethics committee approved this study, and written informed parental consent was obtained. In 273 infants with conjugated hyperbilirubinemia (total bilirubin level ≥ 31.2 µmol/L, with direct bilirubin level > indirect bilirubin level), detailed abdominal US was performed to exclude biliary atresia. Biliary atresia was found in 129 infants and ruled out in 144. A modified triangular cord thickness was measured at the anterior branch of the right portal vein, and a gallbladder classification scheme was identified that incorporated the appearance of the gallbladder and a gallbladder length-to-width ratio of up to 5.2 when the lumen was visualized, as well as HA diameter and liver and spleen size. Reference standard diagnosis was based on results of one or more of the following: surgery, liver biopsy, cholangiography, and clinical follow-up. Area under the receiver operating characteristic curve (AUC) analysis, binary logistic regression analysis, Fisher exact test, and unpaired t test were performed. Results Triangular cord thickness, HA diameter, ratio of gallbladder length to gallbladder width, liver size, and spleen size exhibited statistically significant differences (all P < .05) between the group with biliary atresia and the group without. AUCs of triangular cord thickness, ratio of gallbladder length to width, and HA diameter were 0.952, 0.844, and 0.838, respectively. Logistic regression analysis demonstrated that these three US parameters were significantly associated (all P < .05) with biliary atresia. The combination of triangular cord thickness and gallbladder classification could yield comparable AUCs (0.915 vs 0.933, P = .400) and a higher sensitivity (96.9% vs 92.2%), compared with triangular cord thickness alone. Conclusion By using the combination of modified triangular cord thickness and gallbladder classification scheme, most infants with biliary atresia could be identified. (©) RSNA, 2015.

Alcohol intake and associated risk of major cardiovascular outcomes in women compared with men: a systematic review and meta-analysis of prospective observational studies.

BACKGROUND: The prevalence of alcohol intake is increasing among women in some populations. Alcohol consumption plays an important role in the risk of major cardiovascular outcomes and total mortality. Here, we conducted a meta-analysis to estimate the association between alcohol intake and major cardiovascular outcomes or total mortality in women compared with men. METHODS: We searched the PubMed, Embase, and the Cochrane Library databases for relevant articles published prior to June 2014. Among these potential included prospective studies, the different dose categories of alcohol intake were compared with the lowest alcohol intake or non-drinkers between women and men for the outcomes of major cardiovascular or total mortality. RESULTS: We included 23 prospective studies (18 cohorts) reporting data on 489,696 individuals. The summary relative risk ratio (RRR; female to male) for total mortality was significantly increased with moderate alcohol intake compared with the lowest alcohol intake (RRR, 1.10; 95 % confidence interval [CI]: 1.00-1.21; P = 0.047); no such significance was observed with other levels of alcohol intake (low intake: RRR, 1.07; 95 % CI: 0.98-1.17; P = 0.143; heavy intake: RRR, 1.09; 95 % CI: 0.99-1.21; P = 0.084). There was no evidence of a sex difference in the relative risk for coronary disease, cardiac death, stroke, or ischemic stroke between participants with low to heavy alcohol intake compared with those who never consumed alcohol or had the lowest alcohol intake. CONCLUSIONS: Women with moderate to heavy alcohol intake had a significantly increased risk of total mortality compared with men in multiple subpopulations. Control of alcohol intake should be considered for women, particularly for young women who may be susceptible to binge drinking.

Sonographic features of thyroid nodules that may help distinguish clinically atypical subacute thyroiditis from thyroid malignancy.

OBJECTIVES: The purpose of this study was to evaluate sonographic features for distinguishing clinically atypical subacute thyroiditis from malignant thyroid nodules. METHODS: A total of 165 hypoechoic thyroid nodules without calcification in 135 patients with histologic diagnosis were included in this study. These nodules were classified into 2 groups: a thyroiditis group (55 nodules in 36 patients) and a malignancy group (110 nodules in 99 patients). The sonographic features of the groups were retrospectively reviewed. RESULTS: No significant differences were detected for the variables of marked echogenicity, a taller-than-wide shape, and mixed vascularity. However, a poorly defined margin was detected more frequently in the thyroiditis group than the malignancy group (P < .05); it yielded a high capability for differential diagnosis of atypical subacute thyroiditis, with sensitivity and specificity of 87.3% and 80.9%, respectively. Centripetal reduction echogenicity was observed exclusively in the thyroiditis group, with high specificity (100%) but low sensitivity (21.8%) for atypical subacute thyroiditis diagnosis. All of the thyroiditis nodules with a positive color signal showed noninternal vascularity (negative predictive value, 100%). CONCLUSIONS: There is a considerable overlap between the sonographic features of atypical subacute thyroiditis and thyroid malignancy. However, the margin, echogenicity, and vascularity type are helpful indicators for differential diagnosis of atypical subacute thyroiditis.

Percutaneous radiofrequency ablation of malignant liver tumors with ultrasound and CT fusion imaging guidance.

BACKGROUND: To evaluate the feasibility, accuracy, and utility of sonography (US) and CT fusion imaging guidance for radiofrequency ablation (RFA) of malignant liver tumors not visualized on conventional US. METHODS: Seventy-seven patients with hepatocellular carcinoma and 15 patients with metastatic liver cancer with a total of 136 lesions underwent RFA with US and CT fusion imaging guidance. The mean number of punctures, success rate of a single ablation session, local tumor progression rates, and long-term outcome were evaluated. Treatment efficacy was evaluated by dynamic CT and contrast-enhanced US 1 month after RFA. RESULTS: RFA was technically feasible in all patients, and no major complications occurred. The mean ± SD time needed to synchronize US and CT images was 13.9 ± 11.9 minutes (range, 5-55 minutes). The success rate of a single ablation session was 83.8% (114/136), and tumor residue was present in 7.4% of lesions (10/136). The mean number of treatment sessions was 1.2 ± 0.5 sessions. During follow-up, local tumor progression was observed for 15 (11.9%) lesions. Distant tumor recurrence was found in 51 (55.4%) patients. CONCLUSIONS: US and CT fusion-assisted RFA is a safe and efficacious treatment for patients with hepatocellular carcinoma and metastatic liver cancer.

Natural chalcopyrite mitigates nitrous oxide emissions in sediment from coastal wetlands.

Coastal wetlands are one of the most important natural sources of nitrous oxide (N2O). Previous studies have shown that copper-containing chemicals are able to reduce N2O emissions from these ecosystems. However, these chemicals may harm organisms present in coastal waters and sediment, and disturb the ecological balance of these areas. Here, we first investigated the physiological characteristics and genetic potential of denitrifying bacteria isolated from coastal wetlands. Based on an isolated denitrifier carrying a complete denitrification pathway, we tested the effect of the natural mineral chalcopyrite on N2O production by the bacteria. The results demonstrated that chalcopyrite addition lowers N2O emissions from the bacteria while increasing its N2 production rate. Among the four denitrification genes of the isolate, only nosZ gene expression was significantly upregulated following the addition of 2 mg L-1 chalcopyrite. Furthermore, chalcopyrite was applied to coastal wetland sediments. The N2O flux was significantly reduced in 50-100 mg L-1 chalcopyrite-amended sets relative to the controls. Notably, the dissolved Cu concentration in chalcopyrite-amended sediment remained within the limit set by the National Sewage Treatment Discharge Standard. qPCR and metagenomic analysis revealed that the abundance of N2O-reducing bacteria with the nosZ or nirK + nosZ genotype increased significantly in the chalcopyrite-amended groups relative to the controls, suggesting their active involvement in the reduction of N2O emissions. Our findings offer valuable insights for the use of natural chalcopyrite in large-scale field applications to reduce N2O emissions.

Sorafenib suppresses the rapid progress of hepatocellular carcinoma after insufficient radiofrequency ablation therapy: an experiment in vivo.

BACKGROUND: Radiofrequency ablation (RFA) is a widely applied treatment for hepatocellular carcinoma (HCC), but insufficient RFA can promote rapid progression of the residual tumor through the hypoxia inducible factor-1α (HIF-1α)/vascular endothelial growth factor A (VEGFA) pathway. Although sorafenib has been successfully applied to advanced HCC, the use of sorafenib in residual tumor cells after RFA has rarely been tested. PURPOSE: To evaluate the potential role of sorafenib as an adjunct to RFA to reduce the recurrence rate after insufficient RFA. MATERIAL AND METHODS: Xenograft tumors of SMMC 7721 were created by subcutaneously inoculating nude mice with hepatoma cells (5 × 10(6) cells per mouse). Fourteen days after inoculation, all mice were divided into three groups (control group [sham puncture], RFA group, and RFA combined with sorafenib treatment group) with six mice in each group. Each group was given a different treatment procedure. After treatment, the volume of the tumors was calculated from the resected specimens. The mRNA and protein expression of HIF-1α and VEGFA was quantified by real-time PCR and immunohistochemistry analysis. The micro-vessel density (MVD) was determined by CD34 immunohistochemistry. RESULTS: Real-time PCR and immunohistochemistry analysis showed that, compared to the RFA group, HIF-1α and VEGFA expression were significantly decreased in the group that received RFA combined with sorafenib treatment (P < 0.05). By comparing the control group with the RFA group, we found that insufficient RFA promoted HIF-1α and VEGFA expression (P < 0.05). Similar results were obtained for MVD expression. Additionally, the combination of RFA with sorafenib therapy resulted in a synergistic reduction in tumor growth compared to insufficient RFA and sham puncture (P < 0.05). CONCLUSION: Sorafenib was able to inhibit the expression of HIF-1α and VEGFA, and sorafenib was able to increase time to recurrence when used as an adjunct to RFA.

Percutaneous ultrasound-guided radiofrequency ablation treatment and genetic testing for renal cell carcinoma with Von Hippel-Lindau disease.

Percutaneous ultrasound-guided radiofrequency ablation is increasingly being studied in the treatment of renal tumors. Because percutaneous ultrasound-guided radiofrequency ablation is a minimally invasive and nephron-sparing procedure, it is ideally suited for patients with a single kidney, multiple tumors, or contraindications to conventional surgery. We report on a patient with Von Hippel-Lindau (VHL) disease who had multicentric tumors in the single kidney that was successfully treated with percutaneous ultrasound-guided radiofrequncy ablation. The one-year follow-up showed that there was no local recurrence or metastasis. And genetic testing showed the patient had a T to G heterozygotic missense mutation at nucleotide 515 of VHL gene exon 1.

[Ultrasound-guided compression repair for iatrogenic femoral artery pseudoaneurysm].

OBJECTIVE: To evaluate relative factors affecting the efficiency of ultrasound-guided compression repair in iatrogenic femoral artery pseudoaneurysm. METHODS: Ultrasound-guided manual compression was performed in 42 patients of iatrogenic femoral artery pseudoaneurysm from June 2004 to June 2010. There were 28 male and 14 female patients, with a mean age of (52 ± 5) years. These patients were presented with femoral artery pseudoaneurysm after catheterisation procedure by percutaneous femoral artery puncture and confirmed by color doppler flow image. Ultrasound-guided manual persistent compression with probe was performed at the puncture site between femoral artery and pseudoaneurysm, until completely thrombosis of pseudoaneurysm, whereas the pseudoaneurysm failed to complete closure required surgical repair. RESULTS: Out of 42 patients, 34 patients (81.0%) were successfully treated by compression resulted in completely thrombosis. There were 8 (19.0%) failures conversion to surgery. Factors associated with success were size of pseudoaneurysm (< 25 mm, 25 - 40 mm, > 40 mm; χ(2) = 13.956, P = 0.001), anti-coagulation status (χ(2) = 5.578, P = 0.010), depth of artery break (< 50 mm, 50 - 80 mm, > 80 mm; χ(2) = 14.055, P = 0.001), pseudoaneurysm communicated with common femoral artery, superficial femoral artery and profunda femoral artery (χ(2) = 8.968, P = 0.011), as well as days to presented with pseudoaneurysm (< 3 d, ≥ 3 d; χ(2) = 5.733, P = 0.012). In multivariate Logistic regression analysis, success by compression was associated with size of pseudoaneurysm (WALD = 5.34, P = 0.021) and with depth of artery break (WALD = 4.84, P = 0.028). CONCLUSION: The ultrasound-guided compression repair of iatrogenic femoral artery pseudoaneurysm is safe, convenient, inexpensive and reliable treatment.

[Clinical Study on the Distribution Frequency of Anti-Mur in Southern Fujian Area and Its Cause of Hemolytic Disease of the Newborn].

OBJECTIVE: To investigate the distribution frequency of anti-Mur and the characteristics of hemolytic disease of the newborn by anti-Mur in this region, and to provide basis for guiding clinical safe blood transfusion. METHODS: Anti-Mur detected in blood preparation or transfusion cases from November 2020 to December 2021 in our hospital were retrospectively analyzed, the laboratory test data and relevant clinical data of the patients were statistically analyzed. RESULTS: Among 39 069 cases of blood preparation or transfusion, 345 cases of irregular antibody were detected, including 23 cases of anti-Mur, 7 cases of male, and 4 cases of blood transfusion history. Among the 16 women, 9 had history of only pregnancy, 2 had history of only blood transfusion, and 5 had history of both blood transfusion and pregnancy. The median age of anti-Mur patients was 59(19-78) years old, which was higher than the median age of other positive patients in the same period, 45(1 day-93 year) years old (P<0.05), but there was no significant difference in gender between groups (P＞0.05). A child with hyperbilirubinemia was diagnosed with hemolytic disease of the newborn by anti-Mur after detection of anti-Mur in erythrocyte elution liquid and maternal serum. CONCLUSION: Anti-Mur has important clinical significance in Southern Fujian area, and its generation is closely related to the history of immunization. The age of anti-Mur detected is higher than that of other positive patients. Hemolytic disease of the newborn caused by anti-Mur has severe anemia symptoms and is easy to miss detection, which should be paid more attention to.

Pre-operative breathing training based on video learning reduces emergence delirium in preschool children: A randomized clinical trial.

STUDY OBJECTIVE: Emergence delirium is a common complication in preschool children after general anesthesia and may result in undesirable complications. This study aimed to determine whether breathing training after watching an informative video during the pre-operative visit could reduce the incidence of emergence delirium in preschool children after otorhinolaryngologic surgery under general anesthesia. DESIGN: A single-center, double-blinded, randomized controlled trial. SETTING: Perioperative care. PATIENTS: A total of 170 children undergoing otorhinolaryngologic surgery, aged 3-7 years, ASA physical status I or II were involved. INTERVENTIONS: Patients were randomized to receive breathing training during the pre-operative visit (Training group) or to receive pre-operative visit only (Control group) the day before surgery. MEASUREMENTS: Emergence delirium was measured by the Pediatric Anesthesia Emergence Delirium score during the anesthesia recovery time. Data regarding extubation time and post-anesthesia care unit stay time were collected. MAIN RESULTS: Children who received breathing training during the pre-operative visit had a significantly lower incidence of emergence delirium than those who only underwent the pre-operative visit (10.4% vs. 35.1%, P < 0.001). The awakening time score and the maximum score in the post-anesthesia care unit were significantly lower in the training group compared with the control group [4.4 ± 3.4 vs. 6.9 ± 4.2, P < 0.001 and 5.0 (5.0) vs 7.0 (7.0), P = 0.001, respectively]. We found no differences in the extubation time and post-anesthesia care unit stay time between groups. CONCLUSIONS: We concluded that breathing training based on video learning during the pre-operative visit in preschool children undergoing otorhinolaryngologic surgery could significantly decrease the incidence of emergence delirium. TRIAL REGISTRATION: Chinese Clinical Trial Registry (Reference number: ChiCTR1900026162); registered on September 24, 2019.

Expanding the phylogenetic distribution of cytochrome b-containing methanogenic archaea sheds light on the evolution of methanogenesis.

Methane produced by methanogenic archaea has an important influence on Earth's changing climate. Methanogenic archaea are phylogenetically diverse and widespread in anoxic environments. These microorganisms can be divided into two subgroups based on whether or not they use b-type cytochromes for energy conservation. Methanogens with b-type cytochromes have a wider substrate range and higher growth yields than those without them. To date, methanogens with b-type cytochromes were found exclusively in the phylum "Ca. Halobacteriota" (formerly part of the phylum Euryarchaeota). Here, we present the discovery of metagenome-assembled genomes harboring methyl-coenzyme M reductase genes reconstructed from mesophilic anoxic sediments, together with the previously reported thermophilic "Ca. Methylarchaeum tengchongensis", representing a novel archaeal order, designated the "Ca. Methylarchaeales", of the phylum Thermoproteota (formerly the TACK superphylum). These microorganisms contain genes required for methyl-reducing methanogenesis and the Wood-Ljundahl pathway. Importantly, the genus "Ca. Methanotowutia" of the "Ca. Methylarchaeales" encode a cytochrome b-containing heterodisulfide reductase (HdrDE) and methanophenazine-reducing hydrogenase complex that have similar gene arrangements to those found in methanogenic Methanosarcinales. Our results indicate that members of the "Ca. Methylarchaeales" are methanogens with cytochromes and can conserve energy via membrane-bound electron transport chains. Phylogenetic and amalgamated likelihood estimation analyses indicate that methanogens with cytochrome b-containing electron transfer complexes likely evolved before diversification of Thermoproteota or "Ca. Halobacteriota" in the early Archean Eon. Surveys of public sequence databases suggest that members of the lineage are globally distributed in anoxic sediments and may be important players in the methane cycle.

Recombinant adenovirus-mediated human telomerase reverse transcriptase gene can stimulate cell proliferation and maintain primitive characteristics in bovine mammary gland epithelial cells.

The human telomerase reverse transcriptase (hTERT) gene has been used to stimulate the proliferation of most types of human cells. The present study was designed to evaluate the feasibility and efficiency of adenovirus-mediated hTERT in the proliferation of bovine mammary gland epithelial cells (bMGEs). A plasmid and an adenovirus vector that carried hTERT, namely pEGFP- hTERT and Ad- hTERT, were constructed and transfected into bMGEs, respectively. In order to select the best strategy for stimulating cell proliferation, the adenovirus- and plasmid-mediated hTERT were compared in terms of the positive cloning and transgenic efficiency. The results showed that only Ad- hTERT had high infection efficiency and produced a positive polyclone population (hTERT-bMGEs). The characteristics of the hTERT-bMGEs were investigated with further analysis by reverse transcription-polymerase chain reaction (RT-PCR), western blotting, proliferation assays, and flow cytometry, which showed that hTERT facilitated strong cell proliferation. Real-time quantitative PCR showed a normal level of expression of beta-casein, the caspase-8 and c-myc proto-oncogene, and immunofluorescence demonstrated the properties of the epithelial cells. In conclusion, the adenovirus-mediated hTERT gene could not only extend the cell lifespan, but also maintained the primary characteristics of the cells. It may be possible to extend the use of a wide variety of non-human mammalian cells in this way. This study has provided additional insight into the mechanism of cell proliferation by demonstrating the lack of integration of the adenovirus-mediated hTERT gene into the mammalian genome.

Multipotent differentiation of the EGFP gene transgenic stem cells derived from amniotic fluid of goat at terminal gestational age.

We have isolated stem cells from amniotic fluid of goat at terminal gestational age and transferred the EGFP (enhanced green fluorescent protein) gene into the stem cells previously. The aim of this study was to determine whether the transgenic stem cells have the capability of multipotent differentiation. The transgenic stem cells were induced to differentiate into neurogenic, adipogenic, osteogenic and endothelial cells in vitro. Markers associated with AFS (amniotic fluid-derived stem) cells and the differentiated cells were tested by RT-PCR (reverse transcription-PCR). The results demonstrated that the transgenic AFS cells were capable of self-renewal, a defining property of stem cells. AFS cells were positive for the undifferentiated cell markers, Oct4, Nanog, Sox2 and Hes1, while following differentiation cells expressed markers for neurogenic cells such as astrocyte [GFAP (glial fibrillary acidic protein)] and NSE (neuron-specific enolase), adipogenic cells [LPL+ (lipoprotein lipase+)], osteogenic cells (osteocalcin+ and osteonectin+) and endothelium [CD34+ and eNOS+ (endothelial nitric oxide synthase)]. The results demonstrated that the EGFP gene transgenic AFS cells have the capability of multipotent differentiation, which means that the transgenic AFS cells may be useful in cell-transplantation studies in future.

[Investigation and Analysis of Non-ABO Hemolytic Disease of the Newborn].

OBJECTIVE: To study the serological detection characteristics and antibody specific distribution of hemolytic disease of the newborn (HDN) caused by irregular antibodies through retrospective case analysis. METHODS: A total of 3 047 suspected cases of HDN were submitted by the Neonatal Department of our hospital from January 2014 to December 2019. Non ABO-HDN cases confirmed in our laboratory were taken as the research objects, while some cases of ABO-HDN were randomly selected as control. Disease-causing antibody specificity, serological detection characteristics, total bilirubin change trend and gender ratio of non ABO-HDN patients were explored. RESULTS: Sixty-seven cases of non ABO-HDN were confirmed from the suspected cases of HDN, Among which 45 males and 22 females were detected with the positive rate 1.48% and 0.72%, respectively. The mothers of 65 cases had two or more pregnancies. The detected irregular antibodies were mainly involved with Rh system, MNS system, Kidd system and Lewis system, among which Rh system accounted for 88.07% of the total antibody detection rate. Compared with that of ABO-HDN patients, the total bilirubin of non ABO-HDN patients developed more rapidly with a higher peak and a longer duration (P<0.001). In terms of serological detection, the positive rate of non ABO-HDN direct antibody test was 97.01%, which was higher than 47.00% of ABO-HDN (P<0.001), and the agglutination strength was often ≥ 2+, but there were still weak positive or negative cases of direct antibody test. CONCLUSION: Non ABO-HDN caused by irregular antibodies mostly occurs in fetuses whose mothers experience multiple pregnancies, and the number of males is more than females. The irregular antibodies detected are mainly attributed to Rh system. The peak value of bilirubin in non ABO-HDN patients is higher and lasts longer than that in ABO-HDN patients. Direct antiglobulin test may be used to roughly distinguish ABO-HDN from non ABO-HDN.

Brain functional changes in patients with Crohn's disease: A resting-state fMRI study.

BACKGROUND: Crohn's disease (CD) is a chronic recurrent intestinal inflammatory disease, often accompanied by poor adaptation and excessive stress response. However, the potential neurological mechanisms of these symptoms have not yet been studied in-depth. OBJECTIVE: To investigate alterations in brain activity in patients with Crohn's disease and study the relationship between altered regions and clinical indices. METHODS: A total of 15 CD patients and 26 matched healthy controls were recruited. All participants underwent fMRI scans. The amplitude of low-frequency fluctuations (ALFF) and regional homogeneity (ReHo) assessed differences in spontaneous regional brain activity. Differences between the groups were selected as seeds for functional connectivity (FC) analyses. Correlations between disease duration and ALFF/ReHo/FC values in abnormal regions were analyzed. RESULTS: Patients with CD had significantly higher ALFF values in the left superior frontal gyrus, anterior cingulate cortex, and supplementary motor area, and lower values in the left hippocampus. They also had higher ReHo values in the left anterior cingulate cortex, supplementary motor area, putamen, and the bilateral superior frontal gyri. FC strength in the left precentral and middle temporal gyri was found to be increased when the left superior frontal gyrus was used as the seed point. FC strength was also observed to be increased in the left postcentral, middle frontal gyri, inferior frontal orbital cortex, and right rolandic operculum when the left anterior cingulate cortex was used as the seed point. CONCLUSION: CD demonstrated abnormal neural activity and FC in various regions primarily associated with emotional, pain and cognitive-related functions, which provides more information to further understand the neural mechanisms of the disease.