Microbial communities such as those residing in the human gut are highly diverse and complex, and many with important implications for health and diseases. The effects and functions of these microbial communities are determined not only by their species compositions and diversities but also by the dynamic intra- and inter-cellular states at the transcriptional level. Powerful and scalable technologies capable of acquiring single-microbe-resolution RNA sequencing information in order to achieve a comprehensive understanding of complex microbial communities together with their hosts are therefore utterly needed. Here we report the development and utilization of a droplet-based smRNA-seq (single-microbe RNA sequencing) method capable of identifying large species varieties in human samples, which we name smRandom-seq2. Together with a triple-module computational pipeline designed for the bacteria and bacteriophage sequencing data by smRandom-seq2 in four human gut samples, we established a single-cell level bacterial transcriptional landscape of human gut microbiome, which included 29,742 single microbes and 329 unique species. Distinct adaptive response states among species in Prevotella and Roseburia genera and intrinsic adaptive strategy heterogeneity in Phascolarctobacterium succinatutens were uncovered. Additionally, we identified hundreds of novel host-phage transcriptional activity associations in the human gut microbiome. Our results indicated that smRandom-seq2 is a high-throughput and high-resolution smRNA-seq technique that is highly adaptable to complex microbial communities in real-world situations and promises new perspectives in the understanding of human microbiomes.
Humans ; Gastrointestinal Microbiome/genetics* ; Bacteriophages/physiology* ; High-Throughput Nucleotide Sequencing ; Sequence Analysis, RNA/methods* ; Bacteria/virology*

Neck dissection(ND)is one of the main treatment methods for oral and maxillofacial malignancies.Although ND type is in con-stant improvement,but intraoperative peal-pull-push injury of the accessory nerve,muscle,muscle membrane,fascia and ligament induced shoulder syndrome(SS)is still a common postoperative complication,combined with the influence of radiochemotherapy,not only can cause pain,stiffness,numbness,limited dysfunction of shoulder neck and arm,but also may have serious impact on patient's life quality and phys-ical and mental health.At present,there is still a lack of a systematic evaluation and rehabilitation management program for postoperative SS of oral and maxillofacial malignant tumors.Based on the previous clinical practice and the current available evidence,refer to the relevant lit-erature at home and abroad,the experts in the field of maxillofacial tumor surgery and rehabilitation were invited to discuss,modify and reach a consenusus on the etiology,assessment diagnosis,differential diagnosis,rehabilitation strategy and prevention of SS,in order to provide clinical reference.

Objective To investigate the prevalence of Anisakis infections in coastal marine fishes and awareness of anisakiasis control knowledge among local residents in Yantai City, Shandong Province in 2021, so as to provide insights into formulation of anisakiasis control interventions. Methods Marine fishes were purchased from Shunxin Port, Yantai City, Shandong Province in November 2021, and the presence of Anisakis was detected in different species of fishes and different fish sites. The correlations between body length and weight of marine fish and intensity of Anisakis infections were examined using Spearman’s rank correlation analysis, and the dietary habits and anisakiasis control knowledge were investigated using questionnaire surveys among local residents. Results A total of 201 marine fishes belonging to 20 species were dissected, and Anisakis was detected in 77 marine fishes (38.31%) belonging to 11 species (55.00%), with a mean infection intensity of 45.04 parasites per fish (3 468/77). Spearman’s rank correlation analysis revealed that the body length (r_s = 0.74, P < 0.05) and weight (r_s = 0.79, P < 0.01) of the monkfish correlated positively with the intensity of Anisakis infections, and the body length (r_s = 0.68, P < 0.05) of the flatfish correlated positively with the intensity of Anisakis infections, while no correlations were examined between the body length or weight of other marine fishes and the intensity of Anisakis infections. Of all respondents, 53.38% men and 56.67% women did not know anisakiasis control knowledge at all, and there was a significant difference in the proportion of respondents using separate chopping boards for raw and cooked food from different villages (χ² = 17.89, P < 0.01), while there was an age-specific proportion of respondents with habitats of eating raw or semi-raw seafood (χ² = 28.27, P < 0.01). Conclusions The prevalence and intensity of Anisakis infections were high in coastal marine fishes in Yantai City in 2021, and the awareness of anisakiasis control knowledge was low among local residents. Intensified health education pertaining to anisakiasis control knowledge is recommended to reduce the risk of Anisakis infections.

Humans ; Endoscopic Ultrasound-Guided Fine Needle Aspiration ; Pancreatic Neoplasms ; Pancreas ; Needles

Objective:To investigate whether silencing signal transducer and activator of transcription 6 (STAT6) with siRNA can inhibit eosinophilic inflammation of sinonasal mucosa in a mouse model of eosinophilic chronic rhinosinusitis (ECRS).Methods:The study was conducted from March to September in 2022. Forty-eight female BALB/c mice were randomly divided into four groups: the control group, the Vehicle (transfection reagent)-treated group, the Scramble siRNA (Control siRNA)-treated group, and the STAT6 siRNA-treated group, with twelve mice in each group. An ovalbumin (OVA)-staphylococcal enterotoxin B (SEB)-induced ECRS murine model was established. SiRNA prepared in Lipofectamine was locally administered to the nasal cavity. After administration, samples of the peripheral blood and sinonasal mucosa were collected. Eosinophils in peripheral blood were detected by hematology analyzer. Total and OVA-specific IgE (OVA-sIgE) in serum were detected by enzyme-linked immunosorbent assay (ELISA). Mucosal levels of cytokines and chemokines, including interleukin (IL)-5, IL-17A, interferon-γ (IFN-γ) and eotaxin-1, were also measured using ELISA. Mucosal histological changes of eosinophil infiltration were examined using hematoxylin, and eosin staining, and tissue eosinophil count was performed using a microscope under a high-power field (HPF). Tissue expression of STAT6 and phosphorylated STAT6 (p-STAT6) was detected with the western blot method. Immunofluorescence staining was used to localize the expression of p-STAT6 in sinonasal mucosa. Statistical analysis was conducted using SPSS 18.0 software.Results:Peripheral blood eosinophil count, percentage of peripheral blood eosinophil, total serum IgE level, and serum OVA-sIgE level in the STAT6 siRNA-treated group [(0.318±0.045)×10 3/μl, (3.667±0.479)%, (102.070±13.205) ng/ml, and (38.870±7.352) ng/ml] were significantly different from those of the Vehicle-treated group [(0.532±0.049)×10 3/μl, (6.710±1.061)%, (203.102±29.653) ng/ml, and (74.575±6.432) ng/ml, Z value was -2.56, -2.24, -2.40, and -2.56, respectively, all P<0.05] and Scramble siRNA-treated group [(0.493±0.036)×10 3/μl, (5.858±0.872)%, (189.964±30.042) ng/ml, and (80.935±8.358) ng/ml, Z value was -2.17, -2.08, -2.24, and -2.72, respectively, all P<0.05]. Besides, IL-5 and eotaxin-1 levels in the STAT6 siRNA-treated group [(312.279±34.281) pg/ml and (25.297±4.323) pg/ml] were significantly lower than those in the Vehicle-treated group [(689.667±31.905) pg/ml and (68.278±6.485) pg/ml, Z value was -2.73 and -2.88, respectively, both P<0.01] and Scramble siRNA-treated group [(661.783±42.094) pg/ml and (63.015±7.416) pg/ml, Z value was -2.72 and -2.81, respectively, both P<0.01]. Tissue eosinophil count in sinonasal mucosa was (29.132±4.163)/HPF in the STAT6 siRNA-treated group, and were significantly less than those in the Vehicle-treated group [(78.050±7.912)/HPF, Z=-2.88, P<0.01] and Scramble siRNA-treated group [(73.864±8.671)/HPF, Z=-2.72, P<0.01]. The expression level of STAT6 protein (0.105±0.021) was significantly decreased in the mice treated with STAT6 siRNA compared with PBS, Vehicle, and Scramble siRNA (0.232±0.037, 0.243±0.039, and 0.228±0.032, Z value was -2.25, -2.49, and -2.56, respectively, all P<0.05). Corresponding, p-STAT6 protein level (0.292±0.038) was markedly decreased by the introduction of STAT6 siRNA, the difference was statistically significant as compared with the Vehicle-and Scramble siRNA-treated groups (0.613±0.046 and 0.641±0.050, Z value was -2.81 and -2.88, respectively, both P<0.01). Immunofluorescence staining showed that p-STAT6 was mainly located in the nucleus of nasal epithelial cells and inflammatory cells. The green fluorescence of p-STAT6 expression in sinonasal mucosa in the STAT6 siRNA-treated group was weaker than that in the Vehicle-and Scramble siRNA-treated groups. Conclusion:Intranasal administration of STAT6 siRNA can significantly downregulate STAT6 expression, decrease p-STAT6 level, and prohibit the development of Th2-skewed ECRS.

BACKGROUND: Pneumonia-like primary pulmonary lymphoma (PPL) was commonly misdiagnosed as infectious pneumonia, leading to delayed treatment. The purpose of this study was to establish a computed tomography (CT)-based radiomics model to differentiate pneumonia-like PPL from infectious pneumonia. METHODS: In this retrospective study, 79 patients with pneumonia-like PPL and 176 patients with infectious pneumonia from 12 medical centers were enrolled. Patients from center 1 to center 7 were assigned to the training or validation cohort, and the remaining patients from other centers were used as the external test cohort. Radiomics features were extracted from CT images. A three-step procedure was applied for radiomics feature selection and radiomics signature building, including the inter- and intra-class correlation coefficients (ICCs), a one-way analysis of variance (ANOVA), and least absolute shrinkage and selection operator (LASSO). Univariate and multivariate analyses were used to identify the significant clinicoradiological variables and construct a clinical factor model. Two radiologists reviewed the CT images for the external test set. Performance of the radiomics model, clinical factor model, and each radiologist were assessed by receiver operating characteristic, and area under the curve (AUC) was compared. RESULTS: A total of 144 patients (44 with pneumonia-like PPL and 100 infectious pneumonia) were in the training cohort, 38 patients (12 with pneumonia-like PPL and 26 infectious pneumonia) were in the validation cohort, and 73 patients (23 with pneumonia-like PPL and 50 infectious pneumonia) were in the external test cohort. Twenty-three radiomics features were selected to build the radiomics model, which yielded AUCs of 0.95 (95% confidence interval [CI]: 0.94-0.99), 0.93 (95% CI: 0.85-0.98), and 0.94 (95% CI: 0.87-0.99) in the training, validation, and external test cohort, respectively. The AUCs for the two readers and clinical factor model were 0.74 (95% CI: 0.63-0.83), 0.72 (95% CI: 0.62-0.82), and 0.73 (95% CI: 0.62-0.84) in the external test cohort, respectively. The radiomics model outperformed both the readers' interpretation and clinical factor model ( P <0.05). CONCLUSIONS The CT-based radiomics model may provide an effective and non-invasive tool to differentiate pneumonia-like PPL from infectious pneumonia, which might provide assistance for clinicians in tailoring precise therapy.
Humans ; Retrospective Studies ; Pneumonia/diagnostic imaging* ; Analysis of Variance ; Tomography, X-Ray Computed ; Lymphoma/diagnostic imaging*

Objective:To study the population distribution of Pomacea spp. in Shandong Province and the risk of angiostrongyliasis cantonensis in the local population, and to provide a basis for scientific prevention and control of related diseases. Methods:From July to December 2021, Yanzhou District of Jining City, Ningyang County of Taian City, and Dongying District of Dongying City were selected as surveillance sites to investigate the population and distribution range of Pomacea spp., live snail samples were collected for morphological and genetic identification, and Pomacea spp. infected with the larva of Angiostrongylus cantonensis was detected by lung test. At the same time, sentinel hospital case surveillance was carried out in Yanzhou District, Jining City, and questionnaire was used to study the local residents' awareness of angiostrongyliasis cantonensis and their personal health behaviors. Results:A total of 312 live snail samples were collected. After morphological identification, they were all Pomacea spp.. After gene sequencing, two populations of Pomacea canaliculata and Pomacea maculata were found. No positive snails infected with Angiostrongylus cantonensis were found. A total of 126 patients with headache as the main neurological symptom were admitted to the sentinel hospital, but there were no monitoring cases that met the inclusion criteria. Among the survey population, 48.38% (134/277) of the respondents had heard of angiostrongyliasis cantonensis, 44.77% (124/277) knew that eating Margarya melanioides might cause angiostrongyliasis cantonensis, and 83.39% (231/277) had no related unhealthy eating behavior. Conclusion:Pomacea spp. is found and reported for the first time in Shandong Province, and there is a risk of population infection with angiostrongyliasis cantonensis.

Objective To evaluate the long-term prognostic value of coronary CT angiography-derived fractional flow reserve(CT-FFR)in elderly patients with coronary heart disease(CHD).Methods A retrospective analysis was performed on 1133 patients with clinically suspected CHD from a prospective observational study based on coronary CTA and CT-FFR at the General Hos-pital of Eastern Theater from April 2018 to March 2019,and 330 elderly CHD patients were even-tually included.According to major adverse cardiovascular events(MACE)occurred or not,295 patients were assigned into non-MACE group and 35 patients into the MACE group.Based on cor-onary CTA data,plaque features were analyzed and CT-FFR values were measured in all lesioned vessels.The relationship of plaque features and CT-FFR with MACE was evaluated by using Cox proportional risk regression model,Kaplan-Meier survival curve,and ROC curve analyses.Results The patients with coronary stenosis(≥50％)or CT-FFR value ≤0.8 had a higher risk of MACE(P＜0.01).Univariate Cox analysis showed that coronary stenosis ≥50％and CT-FFR value 0.8 were risk factors of MACE(P＜0.01).After adjusting confounding factors,multivariate Cox analysis indicated that CT-FFR ≤0.8(HR=17.037,95％CI:5.060-57.358,P=0.000)was only independent predictor for MACE.The risk prediction model based on CT-FFR presented better performance than the model based on coronary CTA stenosis(C-index:0.820 vs 0.696,P=0.000).Conclusion CT-FFR≤0.8 is an important independent predictor for long-term MACE in elderly CHD patients.Clinical risk stratification based on CT-FFR may optimize prognostic man-agement strategies in these patients.

Objective:To investigate the effect of image quality, degree of stenosis, calcification, and their first-order interactions on diagnostic performance of coronary computed tomography (CT) angiography-derived fractional flow reserve (CT-FFR).Methods:This is a reanalysis of data from a multi-center retrospective cross-sectional study of CT-FFR in China. A total of 522 patients with suspected or known coronary heart disease [mean age: 61.6 (34.0-83.0) years, 71.8% (354/493) were male] from 11 medical centers including the General Hospital of Eastern Theater Command from May 2015 to October 2019 were enrolled. All patients underwent coronary CT angiography (CCTA), CT-FFR, and invasive FFR examination. Subjective image quality scores of target vessels were recorded on CCTA images, and stenosis was visually assessed at the lesion level. Calcification arc and calcification remodeling index (CRI) were recorded for each lesion. Sensitivity, specificity, accuracy, positive predictive value (PPV), and negative predictive value (NPV) were compared. Two-way analysis of variance was used to analyze the first-order interaction effects of image quality, degree of stenosis, and calcification.Results:A total of 493 patients with 629 lesions with invasive FFR as a reference were included in the study. The overall sensitivity, specificity, and accuracy of CT-FFR were 80.4%, 93.8%, and 88.6%, respectively. The specificity (95.0% vs. 87.3%, χ2=4.11, P=0.043); accuracy (90.1% vs. 81.9%, χ2=6.22, P=0.013); and NPV (89.7% vs. 80.9%, χ2=4.25, P=0.039) of the group with image quality ≥3 was higher than the group with image quality <3. The degree of stenosis affected the sensitivity, PPV, and NPV of CT-FFR and the calcification arc affected the specificity of CT-FFR (all P>0.05). The specificity (95.8% vs. 90.5%, χ2=4.23, P=0.040); accuracy (91.0% vs. 86.1%, χ2=4.01, P=0.045); and NPV (91.1% vs. 83.8%, χ2=5.10, P=0.024) of the group with CRI<1 were higher than that of the group with CRI≥1. In the subgroup of mild and severe stenosis, no calcification, and CRI<1, the accuracy of CT-FFR with image quality ≥3 points were higher than that with image quality <3 points. The accuracy of CT-FFR in the moderate stenosis group was mainly affected by CRI; the accuracy of CT-FFR in the group with CRI<1 was higher than that in the group with CRI≥1 (after Bonferroni correction, P values between groups were statistically significant). Conclusion:Subjective image quality, degree of stenosis, calcification of lesions, and their first-order interactions can all negatively affect the diagnostic performance of CT-FFR.

Objective:To explore the efficacy and safety of intraarterial microguidewire electrocoagulation in arterial aneurysms.Methods:(1) SilverSpeed, a kind of microguidewire used in clinical intravascular treatment for intracranial aneurysms, was used to conduct in vitro electrolysis gas generation experiment with isolated arterial blood of anticoagulant New Zealand white rabbits as medium, and thrombus attachment on the surface of microguidewire was observed under scanning electron microscope. (2) Rabbit common carotid artery aneurysm models were established by using vein bag transplantation method, and divided into microguidewire electrocoagulation treatment groups ( n=40) and blank control group ( n=10). The number of closured tumor cavity and the quality of formed thrombus were observed after electrocoagulation simulation treatment with SilverSpeed microguidewire (charging at 6, 9, 12, 15, and 18 V voltage, respectively for 1, 3, 6, 9, 12, and 15 min). DSA was used to observe whether there was ruptured aneurysms or thrombosis of parent artery. Twelve h later, head MRI diffusion weighted sequence scan was performed to detect whether there were new cerebral ischemia foci in the distal cerebral blood supply area of the parent artery. DSA was performed again 6 months after surgery to observe whether the aneurysms recurred. Results:(1) Electrolytic gas generation experiment results showed that bubbles were generated after electrification of SilverSpeed microguidewire; the higher the voltage, the more severe the reaction. Scanning electron microscope showed that thrombus attached to the surface of the microguidewire after electrification in isolated blood; and the higher the voltage, the denser the thrombus. (2) Under the same charging time, the higher the voltage, the larger the number of closured tumor cavity in rabbits of the microguidewire electrocoagulation treatment groups. Under the same voltage, the longer the charging time, the better the quality of thrombosis. Ischemic events occurred only in the microguidewire electrocoagulation treatment group with voltage>9 V, and the charging duration was not associated with the incidence of embolic events. When the voltage was 15 V, 2 experimental rabbits died due to aneurysm rupture 3 min after electrification. When the voltage was 18 V, 4 experimental rabbits died of cardiac arrest 9 min after electrification, and another 2 rabbits died of aneurysm rupture 6 min after electrification.Conclusions:High voltage is the main cause of adverse events in the microguidewire electrocoagulation treatment of aneurysms. After setting the appropriate voltage, prolonging the electrification time can improve the electrocoagulation effect without increasing the safety risk.