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 investigate the status and influencing factors of substantial weight retention in patients with gestational diabetes mellitus (GDM) in early postpartum.Methods:This cross-sectional study employed a convenience sampling method to select 373 patients with GDM who underwent a 6-week postpartum health check at the Postpartum Health Clinic of Women′s Hospital of Nanjing Medical University from February to July in 2023. According to postpartum weight retention (PPWR) on the survey day, the participants were divided into a substantial PPWR group (≥5 kg, 139 cases) and a non-substantial PPWR group (<5 kg, 234 cases). Independent sample t-tests and χ2 tests were used to compare differences in demographic data, clinical data, and biochemical indicators between the two groups. Binary logistic regression was used to analyze the influencing factors of early substantial postpartum weight retention in patients with GDM. Results:A total of 37.27% (139/373) of GDM patients experienced substantial postpartum weight retention at 6 weeks postpartum. The rates of diabetes in family history, excessive gestational weight gain, gestational hypertension, and infant neonatal intensive care unit (NICU) admission in substantial PPWR group were all significantly higher than those in the non-substantial PPWR group (32.37% vs 23.08%, 73.38% vs 20.09%, 13.67% vs 5.56%, 21.58% vs 9.83%) (all P<0.05); but the proportion of overweight or obese before pregnancy and postpartum self-monitoring of blood glucose were both lower in the substantial PPWR group than those in the non-substantial PPWR group (25.18% vs 41.03%, 17.99% vs 27.78%)(all P<0.05). Binary logistic regression indicated that family history of diabetes ( OR=2.826, 95% CI: 1.517-5.265), excessive gestational weight gain ( OR=19.240, 95% CI: 10.360-35.732), and infant NICU admission ( OR=2.447, 95% CI: 1.150-5.205) were positively related to the occurrence of substantial weight retention in patients with GDM in early postpartum, whereas pre-pregnancy overweight or obesity ( OR=0.189, 95% CI: 0.100-0.360) was negatively correlated to substantial weight retention in patients with GDM in early postpartum (all P<0.05). Conclusion:Substantial weight retention is prevalent among GDM patients in early postpartum, which is significantly influenced by family history of diabetes, pre-pregnancy overweight or obesity, excessive gestational weight gain, and infant NICU admission history.

Objective To investigate the effect of ventilation with different tidal volumes on respiratory mechanics and haemodynamics in children with severe pneumonia.Methods A total of 104 children with severe pneumonia were divided into the control group and the observation group using a random number table method,with 52 cases in each group.Both groups underwent mechanical ventilation treatment.The control group was treated with conventional tidal volume of 10-12 mL/kg,while the observation group was given a low tidal volume of 6-8 mL/kg.The respiratory mechanics indicators,hemodynamic indicators and inflammatory factor indicators before and after treatment were compared between the two groups.Results After treatment,the airway resistance(AR)and airway closing pressure(AOP)were lower in the observation group than those of the control group(P＜0.05),and pulmonary dynamic compliance(Cdyn)was higher in the observation group than that of the control group(P＜0.05).There were no significant differences in heart rate(HR),mean arterial pressure(MAP)and central venous pressure(CVP)after treatment between the two groups(P＜0.05).After treatment,the levels of interleukin(IL)-6,IL-8 and tumor necrosis factor-α(TNF-α)were all lower in the observation group than those in the control group(P＜0.05).Conclusion Small tidal volume mechanical ventilation is effective in improving the respiratory mechanics of children with severe pneumonia without causing large fluctuations in their hemodynamics.At the same time,it can reduce the level of inflammatory factors in the organism.

Objective To investigate the effect of ventilation with different tidal volumes on respiratory mechanics and haemodynamics in children with severe pneumonia.Methods A total of 104 children with severe pneumonia were divided into the control group and the observation group using a random number table method,with 52 cases in each group.Both groups underwent mechanical ventilation treatment.The control group was treated with conventional tidal volume of 10-12 mL/kg,while the observation group was given a low tidal volume of 6-8 mL/kg.The respiratory mechanics indicators,hemodynamic indicators and inflammatory factor indicators before and after treatment were compared between the two groups.Results After treatment,the airway resistance(AR)and airway closing pressure(AOP)were lower in the observation group than those of the control group(P＜0.05),and pulmonary dynamic compliance(Cdyn)was higher in the observation group than that of the control group(P＜0.05).There were no significant differences in heart rate(HR),mean arterial pressure(MAP)and central venous pressure(CVP)after treatment between the two groups(P＜0.05).After treatment,the levels of interleukin(IL)-6,IL-8 and tumor necrosis factor-α(TNF-α)were all lower in the observation group than those in the control group(P＜0.05).Conclusion Small tidal volume mechanical ventilation is effective in improving the respiratory mechanics of children with severe pneumonia without causing large fluctuations in their hemodynamics.At the same time,it can reduce the level of inflammatory factors in the organism.

Objective:To investigate the status and influencing factors of substantial weight retention in patients with gestational diabetes mellitus (GDM) in early postpartum.Methods:This cross-sectional study employed a convenience sampling method to select 373 patients with GDM who underwent a 6-week postpartum health check at the Postpartum Health Clinic of Women′s Hospital of Nanjing Medical University from February to July in 2023. According to postpartum weight retention (PPWR) on the survey day, the participants were divided into a substantial PPWR group (≥5 kg, 139 cases) and a non-substantial PPWR group (<5 kg, 234 cases). Independent sample t-tests and χ2 tests were used to compare differences in demographic data, clinical data, and biochemical indicators between the two groups. Binary logistic regression was used to analyze the influencing factors of early substantial postpartum weight retention in patients with GDM. Results:A total of 37.27% (139/373) of GDM patients experienced substantial postpartum weight retention at 6 weeks postpartum. The rates of diabetes in family history, excessive gestational weight gain, gestational hypertension, and infant neonatal intensive care unit (NICU) admission in substantial PPWR group were all significantly higher than those in the non-substantial PPWR group (32.37% vs 23.08%, 73.38% vs 20.09%, 13.67% vs 5.56%, 21.58% vs 9.83%) (all P<0.05); but the proportion of overweight or obese before pregnancy and postpartum self-monitoring of blood glucose were both lower in the substantial PPWR group than those in the non-substantial PPWR group (25.18% vs 41.03%, 17.99% vs 27.78%)(all P<0.05). Binary logistic regression indicated that family history of diabetes ( OR=2.826, 95% CI: 1.517-5.265), excessive gestational weight gain ( OR=19.240, 95% CI: 10.360-35.732), and infant NICU admission ( OR=2.447, 95% CI: 1.150-5.205) were positively related to the occurrence of substantial weight retention in patients with GDM in early postpartum, whereas pre-pregnancy overweight or obesity ( OR=0.189, 95% CI: 0.100-0.360) was negatively correlated to substantial weight retention in patients with GDM in early postpartum (all P<0.05). Conclusion:Substantial weight retention is prevalent among GDM patients in early postpartum, which is significantly influenced by family history of diabetes, pre-pregnancy overweight or obesity, excessive gestational weight gain, and infant NICU admission history.

Objective To investigate the effect and mechanism of Di'ao Xinxuekang(hereinafter referred to as Xinxuekang)combined with Simvastatin on atherosclerosis(AS)mice.Methods Eight C57BL/6J mice were used as control group,and 32 ApoE-/-mice were randomly divided into model group,Xinxuekang group(160 mg·kg-1),Simvastatin group(1.3 mg·kg-1)and combined treatment group(Xinxuekang 160 mg·kg-1+Simvastatin 1.3 mg·kg-1),with eight mice in each group.The control group was fed with conventional diet,and the other four groups were fed with high-fat diet.At the same time,each administration group was given intragastric administration according to the above dose,and the volume of intragastric administration was 10 mL·kg-1,once a day for 18 weeks.After administration,the levels of serum total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C)and high density lipoprotein cholesterol(HDL-C)were detected.Oil red O staining was used to observe the formation of aortic plaque and liver lipid accumulation in mice.Serum PCSK9 level was detected by ELISA.The mRNA and protein expression levels of LDLR,HNF1α and SREBP2 in liver tissues were detected by qRT-PCR and Western Blot.Results(1)Compared with the control group,the levels of serum TC,TG and LDL-C in the model group were significantly increased(P＜0.01),and the level of HDL-C was significantly decreased(P＜0.01).The percentage of aortic root plaque area,the percentage of total aortic plaque area and the percentage of liver lipid droplet area were significantly increased(P＜0.01).The mRNA and protein expression levels of LDLR in liver tissue were significantly decreased(P＜0.01),and the serum PCSK9 level was significantly increased(P＜0.01).The mRNA and protein expression levels of HNF1α and SREBP2 in liver tissues were significantly increased(P＜0.05,P＜0.01).(2)Compared with the model group,the levels of serum TC,TG and LDL-C in the Xinxuekang group and the combined treatment group were significantly decreased(P＜0.05,P＜0.01),and the level of HDL-C was significantly increased(P＜0.05).The level of serum LDL-C in Simvastatin group was significantly decreased(P＜0.01).The percentage of aortic root plaque area and the percentage of total aortic plaque area in the Xinxuekang group and the combined treatment group were significantly decreased(P＜0.05,P＜0.01),and the percentage of liver lipid droplet area in each administration group was significantly decreased(P＜0.01).The protein expression level of LDLR in liver tissue of mice in Xinxuekang group and combined treatment group was significantly increased(P＜0.05),the serum PCSK9 level was significantly decreased(P＜0.05,P＜0.01),and the mRNA and protein expression levels of HNF1 α and SREBP2 in liver tissue were significantly decreased(P＜0.05,P＜0.01).(3)Compared with the Simvastatin group,the serum HDL-C level in the combined treatment group was significantly increased(P＜0.05).The percentage of aortic root plaque area and the percentage of liver lipid droplet area were significantly decreased(P＜0.05,P＜0.01).The protein expression level of LDLR in liver tissue was significantly increased(P＜0.01),and the serum PCSK9 level was significantly decreased(P＜0.01).The expression levels of HNF1α protein and SREBP2 mRNA in liver tissues were significantly decreased(P＜0.05,P＜0.01).Conclusion Xinxuekang may play a synergistic effect on lipid-lowering and anti-AS effects of Simvastatin by inhibiting the expressions of SREBP2 and HNF1α and regulating the PCSK9/LDLR signaling pathway.

Objective:To analyze the distribution and drug resistance of pathogens of bacterial bloodstream infection in patients with hematological diseases in the Department of Hematology of Tianjin Medical University General Hospital, and to provide etiological data for clinical empirical anti-infection treatment.Methods:A retrospective analysis was conducted on the general clinical information, pathogenic bacteria and drug susceptibility test results of patients with hematological diseases diagnosed with bacterial bloodstream infection by menstrual blood culture in our center from January 2016 to December 2022.Results:Patients included 498 inpatients, with a total of 639 bacterial strains. Among the patients, 86.9% patients had malignancies, and 76.7% had agranulocytosis. Symptoms of concurrent infections, including those of the respiratory tract, oral mucosa, skin and soft tissues, and abdominal sources were observed in 68.3% patients. Gram-negative bacteria (G -) accounted for 79.0% of the isolated bacteria, and gram-positive bacteria (G +) accounted for 21.0%. The top five isolated pathogens were Klebsiella pneumoniae (22.5%), Escherichia coli (20.8%), Pseudomonas aeruginosa (15.0%), Enterococcus faecium (5.5%), and Stenotrophomonas maltophilum (5.0%). Escherichia coli exhibited a decreasing trend of resistance to quinolones, cephalosporins, and carbapenems. Klebsiella pneumoniae exhibited increasing rates of resistance to quinolones and cephalosporins between 2016 and 2018, but the rated decreased after 2019. The resistance rate to carbapenems exhibited by Pseudomonas aeruginosa was approximately 20%. Carbapenem-resistant strains of Pseudomonas aeruginosa strains were first detected in 2017, with a peak resistance rate of 35.7%, detected in 2019. A 60.0% resistance rate to methicillin was observed in methicillin-resistant coagulase-negative staphylococci (MRCNS), and one case of linezolid-resistant MRCNS was detected. Conclusions:Pathogenic bacteria of bacterial bloodstream infections were widely distributed in our center, and precautions are warranted against carbapenem resistant P. aeruginosa and Klebsiella pneumoniae.

Objective To evaluate the polishing finish of four different polishing tools.Methods Fifty specimens were selected from suitable zirconia and randomly divided into 5 groups with 10 specimens in each group.Group A was the negative control group;Group B was the EVE group;Group C was the SHOFU group;Group D was the Toboom group,and Group E was the NAIS group.All specimens were subjected to surface roughness measurements after each level of treatment.One specimen in each group was randomly selected after each level of treatment and put into the vacuum coater for surface gold spraying,and then put into the scanning electron microscope for surface morphology observation after vacuuming.Results Final Ra value:The negative group((1.677±0.066)μm)＞SHOFU group((0.357±0.037)μm)＞EVE group((0.248±0.051)μm)＞Toboom group((0.115±0.039)μm)and NAIS group((0.123±0.029)μm).Scanning electron microscope observation showed that the deepest and longest scratches were left on the surface of the specimen with the diamond needle,and the surface of the specimen was gradually smoothed by using the polishing kit to treat the surfacein turn.The surface of the specimens polished by NAIS group and Toboom group had the least and shallow scratches under the mirror.Conclu-sion Toboom and NAIS zirconia polishing tools provide the best polishing results.

Objective:To preliminarily investigate the effect of circIGF2BP3 on autophagy in photoaged dermal fibroblasts.Methods:Human dermal fibroblasts were isolated from circumcised foreskin tissues from 6 children in the Department of Urological Surgery, Third Affiliated Hospital of Sun Yat-sen University. An ultraviolet A (UVA) -induced photoaged human dermal fibroblast model (UVA radiation group) was established by repeated UVA radiation at a dose of 10 J/cm 2 for 14 consecutive days, and human dermal fibroblasts receiving no treatment served as control group. The photoaged cell model was verified by β-galactosidase staining, Western blot analysis for determining P21 protein expression, and cell counting kit-8 (CCK8) assay for evaluating cell viability. Moreover, Western blot analysis was performed to determine the protein expression of autophagy-related proteins P62, microtubule-associated protein 1 light chain 3 (LC3) -Ⅰand LC3-Ⅱ in photoaged human dermal fibroblasts, and real-time quantitative RCR (qRT-PCR) to verify the differential expression of circIGF2BP3 between photoaged and normal human dermal fibroblasts. Furthermore, circIGF2BP3 was biologically annotated. Some cultured primary human dermal fibroblasts were divided into 4 groups: empty vector group transfected with an empty vector, UVA + empty vector group transfected with an empty vector followed by repeated UVA radiation, circIGF2BP3 group transfected with a circIGF2BP3-overexpressing lentiviral vector, UVA + circIGF2BP3 group transfected with a circIGF2BP3-overexpressing lentiviral vector followed by repeated UVA radiation. Western blot analysis was performed to determine the expression of autophagy-related proteins. Statistical analysis was carried out by using t test, one-way analysis of variance and least significant difference- t test. Results:Compared with the control group, the UVA radiation group showed significantly increased proportions of β-galactosidase-positive cells (61.33% ± 5.78% vs. 6.37% ± 0.32%, t = 9.49, P < 0.01) and P21 expression (1.25 ± 0.03 vs. 1.00 ± 0.05, t = 4.26, P < 0.05), but significantly decreased cell viability (74.33% ± 3.48% vs. 100%, t = 7.38, P < 0.01). Moreover, the P62 expression and LC3-Ⅱ/Ⅰ ratio were significantly higher in the UVA radiation group than in the control group (both P < 0.05). The relative expression of circIGF2BP3 was 0.72 ± 0.04 in the photoaged human dermal fibroblasts, which was significantly lower than that in the normal human dermal fibroblasts (1.00 ± 0.03, t = 5.46, P < 0.01). The P62 expression and LC3-Ⅱ/Ⅰ ratio were significantly lower in the circIGF2BP3 group (0.60 ± 0.01, 0.71 ± 0.01, respectively) than in the empty vector group (1.00 ± 0.02, 1.00 ± 0.01, t = 16.25, 2.75, P < 0.01, < 0.05, respectively), and lower in the UVA + circIGF2BP3 group (1.05 ± 0.02, 2.04 ± 0.05, respectively) than in the UVA + empty vector group (1.31 ± 0.02, 2.72 ± 0.14, t = 10.493, 6.472, respectively, both P < 0.01) . Conclusion:circIGF2BP3 can regulate autophagy in UVA-induced photoaged dermal fibroblasts, which provides a new potential therapeutic target for the prevention and treatment of skin photoaging.

Objective To investigate the effects of human leukocyte-associated antigen-G (HLA-G) expression in silencing trophoblast cell line JEG-3 under normal and hypoxic conditions on invasion and proliferation of JEG-3 cells. Methods Inhibition of HLA-G expression in JEG-3 cells by transfection of small interfering RNA (siRNA),the transfected JEG-3 cells were divided into 4 groups: normoxia control group, hypoxia control group, normoxia inhibition group and hypoxia inhibition group. The levels of HLA-G mRNA and protein in 4 groups of cells were detected by real-time quantitive PCR and western blot. The proliferation activity and invasion ability of 4 groups of cells were determined by methylthiazolyl tetrazolium (MTT) assay and invasion assay.Results (1) Real-time quantitive PCR technology showed: the level of HLA-G mRNA in the hypoxic inhibition group (0.220±0.050) was significantly different (P<0.05), when compared with that in the hypoxic control group (0.630±0.030) and normoxic inhibition group (0.400± 0.020). (2) Western blot analysis showed: the expression level of HLA-G protein in the hypoxic inhibition group was 0.260±0.010, statistically different from that in the hypoxic control group (0.850±0.100) and the normoxic inhibition group (0.560±0.020; P<0.05).(3) MTT showed: proliferative activity of JEG-3 cells in the normoxic inhibition group was 0.490 ± 0.070, the ability of cell proliferation was reduced. When compared with that in the normoxic control group (0.850±0.050), the differences was statistically significant (P<0.05). The proliferative activity of JEG-3 cells in the hypoxic inhibition group (0.330±0.070) was lower than that in the normoxic inhibition group (0.490±0.070), and there was a significant difference (P<0.05). (4) Invasion assay showed: compared with the normoxic control group (98±7), the invasive ability of JEG-3 cells in the normoxic inhibition group (73 ± 7) was weakened, and the difference was statistically significant (P<0.05). The number of transmembrane cells (52±11) of JEG-3 cells in the hypoxic inhibition group was lower than that in the hypoxic control group (72±7), and the difference was statistically significant (P<0.05). Compared with the normoxic inhibition group, the invasion ability of JEG-3 cells in the hypoxic inhibition group decreased, and the difference was statistically significant (P<0.05). Conclusion Under hypoxia, using siRNA technology to down-regulate the expression of HLA-G may affect the proliferation and invasion ability of trophoblast cells, which may be involved in the occurrence of hypertensive disorder of pregnancy.