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*

Objective:To study the effect of anti-melanoma differentiation-related gene 5(MDA5) antibody positive dermatomyositis on the heart of patients.Methods:A total of 71 patients with dermatomyositis diagnosed in Shandong Provincial Hospital Affiliated to Shandong First Medical University from January 1, 2014 to December 31, 2019 were enrolled as the sample group, including anti-MDA5 (+) group( n=28); anti-MDA5(-) groups( n=43). Electrocardiogram and echocardiography were performed in the sample group and the control group. The electrocardiogram, echocardiography and other relevant clinical data of the anti-MDA5 (+) group, anti-MDA5 (-) group and the healthy control group were retrospectively analyzed. The logistic regression analysis model was used to analyze the related factors influencing cardiac involvement in anti-MDA5 (+) patients. Results:In the anti-MDA5 (+) group, more than half of the patients showed elevated levels of lactate dehydrogenase (21/28, 75%) and α-hydroxybutyrate dehydrogenase (16/28, 57%), and 11%(3/28) showed elevated levels of creatine kinase isoenzyme and myoglobin. Compared with the anti-MDA5 (-) group, the white blood cell count in the blood routine of the anti-MDA5 (+) group [5.2 (4.0, 6.5) ×10 9/L vs. 7.8 (5.6, 10.6)×10 9/L, Z=-3.447, P=0.001], creatine kinase [62.5 (29.3, 108.3) U/L vs. 481.0 (179.0, 2 738.0) U/L, Z=-5.895, P<0.001], lactate dehydrogenase [313.0 (239.0, 362.0) U/L vs. 448.0 (291.0, 542.0) U/L, Z=-3.236, P<0.001], creatine kinase isoenzyme [1.9 (1.1, 3.9)ng/ml vs. 17.7 (4.0, 67.2) ng/ml, Z=-4.724, P<0.001], myoglobin [28.2 (20.0, 43.0) ng/ml vs. 307.4 (48.1, 612.2) ng/ml, Z=-4.800, P<0.001]. Electrocardiogram analysis showed that QRS axis [33.5±265.9 vs. 46.9±22.4, t=-2.900, P=0.004], SV1 amplitude [0.7 (0.4, 0.9) vs. 0.9 (0.7, 1.0), Z=-2.148, P=0.023] in anti-MDA5 antibody (+) group in anti-MDA5 antibody (+) group were lower than anti-MDA5 antibody (-) group. QRS duration [84.0 (78.0, 96.5) vs.92.0 (87.8, 100.5), Z=-2.900, P=0.004], QRS axis [33.5±265.9 vs. 46.9±20.4, Z=-2.32, P=0.023] in the anti-MDA5 antibody (+) group were lower than those in healthy control group. Echocardiographic analysis showed that the E peak of anti-MDA5 (+) group [63.0 (52.5, 69.5)] was significantly lower than that of anti-MDA5 (-) group [85.0 (68.0, 108.0), Z=-4.926, P<0.001)]and healthy control group [67.0 (62.8, 80.3), Z=-2.897, P=0.004]. The peak A of anti-MDA5 (+) group [65.5 (56.5, 80.0)] was significantly lower than that of anti-MDA5 (-) group [76.0 (65.0, 90.0), Z=-2.631, P=0.011], but higher than that of healthy control group [55.0(51.0, 66.5), Z=-4.550, P<0.001]. There was no significant difference in echocardiographic findi-ngs among the other groups. All patients with anti-MDA5 (+) dermatomyositis had interstitial lung disease (28/28, 100%). Patients with MDA5 antibody (+++) are more likely to have cardiac involvement than patients with MDA5 antibody (++). Conclusion:The results of relevant examinations in anti-MDA5-DM patients suggest that there is damage to myocardial cells and cardiac function.

Objective To explore the impact of suture configuration and fixation type on the biome-chanical strength of rotator cuff repair,using a factorial design study.Methods Sixteen fresh-frozen porcine shoulder samples were randomized into an anchorless double-row suture bridge transosseous su-tures(DS)group,an anchored double-row suture bridge transosseous-equivalent(DE)group,an an-chorless X-BOX construct transosseous sutures(XS)group,and an anchored X-BOX construct transos-seous-equivalent(XE)group,each of four,according to suture configuration(double-row suture bridge,traditional X-BOX construct)and fixation type(suture anchors,transosseous sutures).Then,their fatigue resistance(first-cycle excursion,gap length difference ratio,and the percentage of ex-posed footprints)and the failure strength(the maximum failure load and the re-tear type)were mea-sured using a biomechanical material testing machine.Results Different suture configurations affected failure strength(F=39.559,P<0.001),with the double-row suture bridge groups(693.07±58.35 N,746.76±138.57 N)showing significantly higher failure strength,compared to the traditional X-BOX groups(462.90±18.91 N,421.43±90.76 N).However,the fixation type did not significantly im-pact failure strength(F=1.161,P=0.302).Moreover,the suture configuration influenced the gap differ-ence ratio(F=7.781,P=0.016),but had no significant correlation with other fatigue resistance indica-tors(P>0.05).Meanwhile,failure strength and fatigue resistance were not correlated with fixation type,and the interaction between suture and fixation type(P>0.05).The incidence of failure types for the four suture configurations was as follows:Type I tendon tear:XS>XE>DS=DE;type II tendon tear:DS>XE>XS=DE;fixing material-related failure:DE>DS=XE=XS.Conclusion The failure strength and gap formation ratio in rotator cuff repair under fatigue loading are influenced by suture configuration,whereas no significant association has been observed with respect to fixation method,whether using transosseous sutures or suture anchors.

Traditional Chinese Medicine (TCM), as a treasure of the Chinese nation, plays a significant role in maintaining public health. In 2019, the Central Committee of the Communist Party of China and the State Council proposed for the first time the establishment of a TCM registration and evaluation evidence system that integrates TCM theory, "personal experience" and clinical trials (referred to as the "Three-in-One" System) to promote the inheritance and innovation of TCM. Subsequently, the National Medical Products Administration issued several guiding principles to advance the improvement and implementation of this system. Owing to the complexity of its implementation, there are still differing understandings within the TCM industry regarding the positioning of the "Three-in-One" Registration and Evaluation Evidence System, as well as the connotation and value orientation of the "personal experience." To address this, Academician WANG Qi, President of the TCM Association, China International Exchange and Promotion Association for Medical and Healthcare and TCM master, led a group of academicians, TCM masters, TCM pharmacology experts and clinical TCM experts to convene a "Seminar on Promoting the Implementation of the ′Three-in-One′ Registration and Evaluation Evidence System for Chinese Medicinals." Through extensive discussions, an expert consensus was formed, clarifying the different roles of the TCM theory, "personal experience" and clinical trials within the system. It was further emphasized that the "personal experience" is the core of this system, and its data should be derived from clinical practice scenarios. In the future, the improvement of this system will require collaborative efforts across multiple fields to promote the high-quality development of the Chinese medicinal industry.

Objective To explore the impact of suture configuration and fixation type on the biome-chanical strength of rotator cuff repair,using a factorial design study.Methods Sixteen fresh-frozen porcine shoulder samples were randomized into an anchorless double-row suture bridge transosseous su-tures(DS)group,an anchored double-row suture bridge transosseous-equivalent(DE)group,an an-chorless X-BOX construct transosseous sutures(XS)group,and an anchored X-BOX construct transos-seous-equivalent(XE)group,each of four,according to suture configuration(double-row suture bridge,traditional X-BOX construct)and fixation type(suture anchors,transosseous sutures).Then,their fatigue resistance(first-cycle excursion,gap length difference ratio,and the percentage of ex-posed footprints)and the failure strength(the maximum failure load and the re-tear type)were mea-sured using a biomechanical material testing machine.Results Different suture configurations affected failure strength(F=39.559,P<0.001),with the double-row suture bridge groups(693.07±58.35 N,746.76±138.57 N)showing significantly higher failure strength,compared to the traditional X-BOX groups(462.90±18.91 N,421.43±90.76 N).However,the fixation type did not significantly im-pact failure strength(F=1.161,P=0.302).Moreover,the suture configuration influenced the gap differ-ence ratio(F=7.781,P=0.016),but had no significant correlation with other fatigue resistance indica-tors(P>0.05).Meanwhile,failure strength and fatigue resistance were not correlated with fixation type,and the interaction between suture and fixation type(P>0.05).The incidence of failure types for the four suture configurations was as follows:Type I tendon tear:XS>XE>DS=DE;type II tendon tear:DS>XE>XS=DE;fixing material-related failure:DE>DS=XE=XS.Conclusion The failure strength and gap formation ratio in rotator cuff repair under fatigue loading are influenced by suture configuration,whereas no significant association has been observed with respect to fixation method,whether using transosseous sutures or suture anchors.

Objective:To study the effect of anti-melanoma differentiation-related gene 5(MDA5) antibody positive dermatomyositis on the heart of patients.Methods:A total of 71 patients with dermatomyositis diagnosed in Shandong Provincial Hospital Affiliated to Shandong First Medical University from January 1, 2014 to December 31, 2019 were enrolled as the sample group, including anti-MDA5 (+) group( n=28); anti-MDA5(-) groups( n=43). Electrocardiogram and echocardiography were performed in the sample group and the control group. The electrocardiogram, echocardiography and other relevant clinical data of the anti-MDA5 (+) group, anti-MDA5 (-) group and the healthy control group were retrospectively analyzed. The logistic regression analysis model was used to analyze the related factors influencing cardiac involvement in anti-MDA5 (+) patients. Results:In the anti-MDA5 (+) group, more than half of the patients showed elevated levels of lactate dehydrogenase (21/28, 75%) and α-hydroxybutyrate dehydrogenase (16/28, 57%), and 11%(3/28) showed elevated levels of creatine kinase isoenzyme and myoglobin. Compared with the anti-MDA5 (-) group, the white blood cell count in the blood routine of the anti-MDA5 (+) group [5.2 (4.0, 6.5) ×10 9/L vs. 7.8 (5.6, 10.6)×10 9/L, Z=-3.447, P=0.001], creatine kinase [62.5 (29.3, 108.3) U/L vs. 481.0 (179.0, 2 738.0) U/L, Z=-5.895, P<0.001], lactate dehydrogenase [313.0 (239.0, 362.0) U/L vs. 448.0 (291.0, 542.0) U/L, Z=-3.236, P<0.001], creatine kinase isoenzyme [1.9 (1.1, 3.9)ng/ml vs. 17.7 (4.0, 67.2) ng/ml, Z=-4.724, P<0.001], myoglobin [28.2 (20.0, 43.0) ng/ml vs. 307.4 (48.1, 612.2) ng/ml, Z=-4.800, P<0.001]. Electrocardiogram analysis showed that QRS axis [33.5±265.9 vs. 46.9±22.4, t=-2.900, P=0.004], SV1 amplitude [0.7 (0.4, 0.9) vs. 0.9 (0.7, 1.0), Z=-2.148, P=0.023] in anti-MDA5 antibody (+) group in anti-MDA5 antibody (+) group were lower than anti-MDA5 antibody (-) group. QRS duration [84.0 (78.0, 96.5) vs.92.0 (87.8, 100.5), Z=-2.900, P=0.004], QRS axis [33.5±265.9 vs. 46.9±20.4, Z=-2.32, P=0.023] in the anti-MDA5 antibody (+) group were lower than those in healthy control group. Echocardiographic analysis showed that the E peak of anti-MDA5 (+) group [63.0 (52.5, 69.5)] was significantly lower than that of anti-MDA5 (-) group [85.0 (68.0, 108.0), Z=-4.926, P<0.001)]and healthy control group [67.0 (62.8, 80.3), Z=-2.897, P=0.004]. The peak A of anti-MDA5 (+) group [65.5 (56.5, 80.0)] was significantly lower than that of anti-MDA5 (-) group [76.0 (65.0, 90.0), Z=-2.631, P=0.011], but higher than that of healthy control group [55.0(51.0, 66.5), Z=-4.550, P<0.001]. There was no significant difference in echocardiographic findi-ngs among the other groups. All patients with anti-MDA5 (+) dermatomyositis had interstitial lung disease (28/28, 100%). Patients with MDA5 antibody (+++) are more likely to have cardiac involvement than patients with MDA5 antibody (++). Conclusion:The results of relevant examinations in anti-MDA5-DM patients suggest that there is damage to myocardial cells and cardiac function.

Objective To explore the role and efficacy of VEGF and HGF gene adenovirus vector in promoting angiogenesis in ischemic tissue.Methods 84 Kunming mice were randomly divided into sham group,control group,VEGF group,HGF group and VEGF+HGF group,and the left lower limb ischemia model was established.The blood supply of ischemic tissue was observed by rheometer,and the expression levels of VEGF and HGF in each group were detected by Western Blot and ELISA.Immunohistochemical staining was used to detect angiogenesis(CD31,SMA)in ischemic tissues.Safety was assessed by side effects during treatment in mice.Results After the successful modeling,the blood flow velocity of the left lower limb in each group decreased significantly.On the 7th day after operation,the blood flow of the left lower limb in each group was significantly better than that on the 0th day after operation(P<0.05),and the blood flow of the left lower limb in Ad-VEGF-HGF group was significantly better than that in other groups(P<0.05).On the 28th day after operation,the blood flow of the left lower limb in Ad-VEGF-HGF group gradually stabilized,the blood flow in Ad-VEGF-HGF group was significantly better than that in other groups,and both VEGF group and HGF group were significantly better than the control group(P<0.05).On the 7th,14th,and 28th days following surgery,HGF and VEGF protein levels in the Ad-HGF,Ad-VEGF,and Ad-VEGF-HGF groups were substantially greater than those in the control group(P<0.05).The expression level in the Ad-VEGF-HGF group peaked on the 14th day(all P<0.001)and subsequently declined to preoperative levels on the 28th day after operation.Conclusion Ad-VEGF-HGF gene injection can effectively boost VEGF and HGF protein expression and rapidly reach the relative peak level,encour-aging angiogenesis after lower limb ischemia,increasing blood flow,and improving lower limb circulation.

Objective To investigate the effect of Kaixinsan(KXS,a traditional Chinese medicine formula)for alleviating adriamycin-induced depression-like behaviors in mice bearing breast cancer xenografts and explore the pharmacological mechanism.Methods Forty female BALB/c mice were randomized equally into control group,model group,and low-and high-dose KXS treatment groups,and in the latter 3 groups,mouse models bearing orthotopic breast cancer 4T1 cell xenografts were established and treated with adriamycin along with saline or KXS via gavage.Depression-like behaviors of the mice were assessed using open field test and elevated plus-maze test,and the changes in serum levels of depression-related factors were examined.RNA-seq analysis and transmission electron microscopy were used and ferroptosis-related factors were detected to explore the mechanisms of adriamycin-induced depression and the therapeutic mechanism of KXS.The results were verified in SH-SY5Y cells using ferroptosis inhibitor Fer-1 as the positive control.Results KXS significantly alleviated depression-like behaviors and depression-related serological changes induced by adriamycin in the mouse models.RNA-seq results suggested that KXS alleviated chemotherapy-induced depression by regulating oxidative stress,lipid metabolism and iron ion binding in the prefrontal cortex.Pathological analysis and detection of ferroptosis-related factors showed that KXS significantly reduced ferroptosis in the prefrontal cortex of adriamycin-treated mice.In SH-SY5Y cells,both KXS-medicated serum and the ferroptosis inhibitor were capable of attenuating adriamycin-induced cell ferroptosis.Conclusion KXS alleviates adriamycin-induced depression-like behaviors in mice by reducing ferroptosis in the prefrontal cortex of breast cancer-bearing mice.

Objective To investigate the effect of Kaixinsan(KXS,a traditional Chinese medicine formula)for alleviating adriamycin-induced depression-like behaviors in mice bearing breast cancer xenografts and explore the pharmacological mechanism.Methods Forty female BALB/c mice were randomized equally into control group,model group,and low-and high-dose KXS treatment groups,and in the latter 3 groups,mouse models bearing orthotopic breast cancer 4T1 cell xenografts were established and treated with adriamycin along with saline or KXS via gavage.Depression-like behaviors of the mice were assessed using open field test and elevated plus-maze test,and the changes in serum levels of depression-related factors were examined.RNA-seq analysis and transmission electron microscopy were used and ferroptosis-related factors were detected to explore the mechanisms of adriamycin-induced depression and the therapeutic mechanism of KXS.The results were verified in SH-SY5Y cells using ferroptosis inhibitor Fer-1 as the positive control.Results KXS significantly alleviated depression-like behaviors and depression-related serological changes induced by adriamycin in the mouse models.RNA-seq results suggested that KXS alleviated chemotherapy-induced depression by regulating oxidative stress,lipid metabolism and iron ion binding in the prefrontal cortex.Pathological analysis and detection of ferroptosis-related factors showed that KXS significantly reduced ferroptosis in the prefrontal cortex of adriamycin-treated mice.In SH-SY5Y cells,both KXS-medicated serum and the ferroptosis inhibitor were capable of attenuating adriamycin-induced cell ferroptosis.Conclusion KXS alleviates adriamycin-induced depression-like behaviors in mice by reducing ferroptosis in the prefrontal cortex of breast cancer-bearing mice.

Objective:To explore the onset characteristics and pathophysiological changes of simple liver cyst in Beijing.Methods:A retrospective cohort study was used. The physical examination data of Department of Health Management of Xuanwu Hospital of Capital Medical University for 10 years from January 1, 2012 to December 31, 2021 were analyzed. Selected clinical data of 37 389 subjects with 2 or more repeated ultrasound examinations, including 17 759 males, 19 630 females, aged (44.4±16.2) years, ranged from 22-103 years. 3 431 cases hepatic cyst were confirmed by repeated ultrasound examination, the data of the liver cyst formation after the same physical examination were the study group ( n=3 431), and the data before cyst formation were the control group ( n=3 431). The observation indicators included: (1) the epidemiological characteristics of liver cysts; (2) the age distribution of the incidence of liver cysts; (3) the gender distribution of the incidence of liver cysts; (4) the pathophysiological changes of liver cysts.Measurement data of normal distribution were expressed as mean±standard deviation ( ± s). The measurement data of skewed distribution were expressed as M ( Q1, Q3), using Wilcoxon signed rank sum test for comparing groups and chi-square test for comparing count data. The factors associated with hepatic cyst pathogenesis were summarized by multivariate Logistic regression. Results:The overall incidence of hepatic cysts was 9.18%, 9.78% in males, 8.63% in females, and the incidence of males was greater than that of females. The incidence of males over 70 to 79 years old decreased slightly, and the incidence in males and females in the other age groups increased with age.Results of multivariate Logistic analysis showed age ( OR=1.01, 95% CI: 1.01-1.02, P<0.01), waist circumference( OR=1.02, 95% CI: 1.01-1.02, P<0.01), systolic blood pressure ( OR=1.00, 95% CI: 0.99-1.00, P=0.013), ALT ( OR=0.99, 95% CI: 0.98-1.00, P<0.01), AST ( OR=1.03, 95% CI: 1.02-1.04, P<0.01), triglyceride lipids ( OR=0.90, 95% CI: 0.86-0.95, P<0.01), HDL( OR=0.71, 95% CI: 0.60-0.83, P<0.01), uric acid ( OR=1.00, 95% CI: 1.00-1.00, P<0.01), creatinine ( OR=0.99, 95% CI: 0.99-0.99, P<0.01) were the factors influencing the occurrence of liver cysts. Conclusions:The incidence of liver cysts increased linearly with age, and the incidence of males was greater than that of females. Age, waist circumference, systolic blood pressure, ALT, AST, triglycerides, HDL, uric acid, and creatinine may interact with the occurrence and development of liver cysts.