Diabetic kidney disease (DKD) is one of the leading causes of end stage renal disease. Despite recent therapies, mortality due to DKD and resources spent on healthcare are important problems. Thus, appropriate markers are needed to predict renal outcomes. Therefore, we investigated the role of peripheral perfusion as an indicator for renal events in patients with type 2 diabetes mellitus. This retrospective cohort study included 566 patients who were admitted to Matsushita Memorial Hospital in Osaka, Japan for type 2 diabetes mellitus. Peripheral perfusion was assessed using perfusion index (PI), which represents the level of circulation through peripheral tissues and was measured on each toe using a Masimo SET Radical-7  instrument. The duration of follow up was 3.0 years. The median age of patients was 70 years (IQR range: 61–77 years) and median PI value was 2.9% (IQR range: 1.8–4.8%). Multiple logistic regression analyses showed that PI (per 1% increase) was associated with an odds ratio of composite of end-stage renal disease (ESRD) and/or doubling of serum creatinine level; n = 40 (odds ratio 0.823 [95% CI: 0.680–0.970]), and composite of ESRD, doubling of serum creatinine level, and renal death and/or cardiovascular death; n = 44 (odds ratio 0.803 [95% CI: 0.665–0.944]). The factors which were statistically significant in univariate analysis and those known to be related factors for renal event were considered simultaneously as independent variables for multiple logistic regression analysis. PI can be a novel indicator for renal events in patients with type 2 diabetes mellitus. DKD develops in approximately 40% of patients with diabetes and is one of the leading causes of end-stage renal disease5. DKD has been reported to be associated with the progression of cardiovascular diseases and increased risk of cardiovascular mortality8. Total deaths due to cardiovascular diseases and infections are highly prevalent in patients with end-stage renal disease (ESRD)9.Despite the advances in renoprotective therapies, such as renin-angiotensin system blockade and sodium-glucose cotransporter-2 inhibitors10. Indeed, normoalbuminuric renal insufficiency is common in patients with type 2 diabetes mellitus11. Interstitial changes such as chronic renal hypoxia might precede glomerular disease without albuminuria, wherein, intrarenal arteriosclerosis might be a main factor for declining eGFR. Further, the peripheral perfusion index (PI) is the ratio of pulsatile blood flow to non-pulsatile blood flow in the monitored tissue and has been shown to reflect changes in peripheral perfusion13. We have recently demonstrated that a low PI value indicates atherosclerosis in clinical care setting14 and diabetic kidney disease in the patients with type 2 disease in the cross sectional study15. However, the association of diminished PI with renal events in patients with type 2 diabetes mellitus has not been reported. Therefore, we aimed to clarify whether PI might be an indicator for renal events including decline in eGFR, cardiovascular death, and progression of albuminuria, in patients with type 2 diabetes mellitus.The progression of DKD involves glomerular hyperfiltration, albuminuria, decline in estimated glomerular filtration rate (eGFR), and ESRD. Patients with type 2 diabetes mellitus present heterogeneous process of DKDThe characteristics of 566 patients enrolled in this study are shown in Table 2. Multiple logistic regression analyses showed that PI (per 1% increase) was associated with an odds ratio of composite of ESRD and/or doubling of serum creatinine level in all patients as well as in patients with eGFR ≥ 60 ml/min/1.73 m2 (odds ratio 0.823 [95% CI: 0.680–0.970] or 0.598 [95% CI: 0.363–0.871]), and composite of ESRD, doubling of serum creatinine level, and renal death and/or cardiovascular death (odds ratio 0.803 [95% CI: 0.665–0.944] or 0.662 [95% CI: 0.458–0.891]). PI value was associated with an odds ratio of progression of albuminuria only in crude model. Moreover, multiple logistic regression analyses (Model 3) in all patients showed that baseline serum creatinine level was associated with an odds ratio of composite of ESRD and/or doubling of serum creatinine level (odds ratio 2.228 [95% CI: 0.987–5.023]), logarithm of baseline UACR was associated with an odds ratio of composite of ESRD, doubling of serum creatinine level, and renal death and/or cardiovascular death (odds ratio 1.678 [95% CI: 1.074–2.642]), and duration of diabetes, logarithm of baseline UACR or history of cardiovascular disease was associated with an odds ratio of progression of albuminuria , respectively. Multiple logistic regression analyses (Model 3) in patients with eGFR ≥ 60 ml/min/1.73 m2 showed that BMI was associated with an odds ratio of composite of ESRD and/or doubling of serum creatinine level (odds ratio 1.169 [95% CI: 1.033–1.330]) and duration of diabetes, baseline serum creatinine level (per 0.1 mg/dl increase), logarithm of baseline UACR or history of cardiovascular disease was associated with an odds ratio of progression of albuminuria , respectively.Table Net reclassification index (NRI) and integrated discrimination improvement (IDI) was derived from logistic regression models (Model 3) with and without logarithm of baseline UACR. Both NRI and IDI suggested that including baseline UACR in Model 3 results in significant improvement in performance. The NRI or IDI was estimated at 0.484 (95% CI: 0.176–0.792), 0.567 (95% CI: 0.210–0.803) and 0.167 (95% CI: −0.012–0.345) or 0.010 (95% CI: 0.0008–0.019), 0.017 (95% CI: 0.004–0.030) and 0.013 (95% CI: 0.003–0.022) in analyses for composite of ESRD and/or doubling of serum creatinine level, composite of end-stage renal disease, doubling of serum creatinine level, renal death and/or cardiovascular death and progression of albuminuria in all patients, respectively.The major finding of our study is that PI, which represents peripheral perfusion, can be a novel indicator for renal events such as composite of ESRD, doubling of serum creatinine level, renal death and/or cardiovascular death in patients with type 2 diabetes mellitus.2 or doubling of the blood creatinine level at a median duration of 15 years after diagnosis15. Patients with ESRD had high prevalence of mortality due to cardiovascular diseases and infections. The overall death rate in patients with ESRD, having creatinine levels >2 mg/dL or those receiving kidney replacement therapy was nearly 20% per year9. In spite of the development of recent therapies, large residual risks of renal events remain. Therefore, an easy and useful tool is required to improve renal outcomes in patients with type 2 diabetes mellitus. The present study suggests that PI could be a useful tool to indicate renal events in patients with type 2 diabetes mellitus.The United Kingdom Prospective Diabetes Study showed that 40% patients developed albuminuria, while 30% patients had eGFR < 60 ml/min/1.73 m15. Low PI value was more associated with eGFR < 60 mL/min per 1.73 m2 than albuminuria, which is consistent with our results. This can be explained by the mechanism and structural changes during decline of eGFR and progression of albuminuria. The initial structural change during DKD is thickening of glomerular basement membrane along with capillary and tubular basement membrane thickening18. Further, glomerular changes include loss of endothelial fenestrations, mesangial matrix expansion, and loss of podocytes17. It has been reported that albuminuria is correlated with mesangial expansion3, which is a marker for glomerular disease and an early clinical indicator of DKD. Diabetic control has been considered as an important factor for prevention of albuminuria16. Previous studies have shown high prevalence of patients with type 2 diabetes mellitus having reduced eGFR without albuminuria. Yokoyama et al.19 reported that proportion of patients with normoalbuminuria was 52% among patients with eGFR < 60 ml/min/1.73 m2. Chronic hypoxia in tubulointerstitium can explain the decline in eGFR without the presence of albuminuria. Chronic hypoxia in tubulointerstitium results in advanced tubulointerstitial lesions besides minor diabetic glomerular lesions21. Some mechanisms can explain development of chronic hypoxia in tubulointerstitium. One of the mechanisms is diminished microcirculation, such as distortion and loss of peritubular capillaries due to intrarenal arteriosclerosis26. These changes may induce a vicious cycle of regional hypoxia and decline in eGFR. Taken together, decrease in eGFR in patients without the presence of albuminuria is mainly associated with atherosclerosis. We have recently reported that low value of PI was strongly correlated with atherosclerosis14. Low value of PI can denote low peripheral perfusion resulting from atherosclerosis. Therefore, PI can be associated with decline in eGFR and not with progression of albuminuria. We have also reported that PI was associated with ankle brachial index14. Lower ankle brachial index has been associated with cardiovascular mortality27. Therefore, PI may be strongly associated with composite renal event including cardiovascular death.In our study, PI was strongly associated with decline in eGFR, composite of ESRD and/or doubling of serum creatinine level. However, multiple regression analysis suggested that PI was not associated with progression of albuminuria. Recently, we have reported the association PI and diabetic kidney disease in the cross sectional studyp = 0.0003). We postulated that PI was also affected by nutrition status. We supposed that PI could be affected by atherosclerosis. However, PI is not correlated with systolic blood pressure, which is consisted with recent report14. This could be explained by the effect of some factors such as duration of hypertension or the effect of antihypertensive medications.In our study, PI is directly correlated with total cholesterol and triglycerides. Moreover, PI is negatively correlated with age and positively correlated with BMI and serum albumin : eGFR = 194 × Cre−1.094 × age−0.287 (ml/min/1.73 m2). For women, the value of eGFR was multiplied by a correction factor of 0.739. ESRD was defined by the initiation of dialysis or eGFR < 15 ml/min/1.73 m2. The history of cardiovascular disease was defined as the patients with the history of myocardial infarction, heart failure, or stroke. Renal death was defined as death with a proximate renal cause. Death due to myocardial infarction, heart failure, or stroke was defined as cardiovascular death. Urine albumin-to-creatinine ratio (UACR) was determined using early morning urine. UACR was measured using an immunoturbidimetric assay. Microalbuminuria was defined by UACR 30 to < 300 mg/g Cr, whereas, macroalbuminuria was defined by UACR ≥ 300 mg/g Cr. Progression of albuminuria was defined as >30% increase in albuminuria, and a change from normoalbuminuria to either microalbuminuria or macroalbuminuria, or from microalbuminuria to macroalbuminuria. Patients for whom PI measurement could not be obtained were excluded from this study. Other exclusion criteria included patients with implanted cardiac pacemakers; arrhythmia, such as paroxysmal atrial fibrillation; amputations in any part of the lower extremities; and patients with malignancy or ESRD at baseline.Diabetes was diagnosed based on the report of the expert committee on the diagnosis and classification of diabetes mellitus30. After bilateral determination, the lower value of PI was considered as a representative for each subject.PI was measured using a Masimo SET Radical-7  instrument. The patients were placed in the supine position. A Masimo pulse oximeter probe was positioned on each toe and connected to Masimo SET Radical-7 machine. The patients were asked to rest for 5 min at the beginning of the procedure. PI was recorded three times: at 20 s, 40 s, and 60 s, after the 5-min rest period. The average of the three values was calculated and used as the reference value. The Masimo SET Radical-7 calculates PI as the ratio between the pulsatile and non-pulsatile components of the light reaching a light-sensitive cell of the pulse oximetry probe. The reliability and reproducibility of PI have been reported elsewhere2. Logistic regression analyses were conducted to examine the effects of PI on composite of ESRD, and/or doubling of serum creatinine level, composite of end-stage renal disease, doubling of serum creatinine level, renal death and/or cardiovascular death, and progression of albuminuria in all patients and in patients with eGFR ≥ 60 ml/min/1.73 m2. The following confounding factors were considered: age, duration of diabetes, body mass index, systolic blood pressure, hemoglobin A1c, total cholesterol and uric acid (Model 1). Model 2 was adjusted for all variables in Model 1 plus creatinine level. Model 3 was adjusted for all variables in Model 2 plus sex, logarithm of UACR, smoking status, history of cardiovascular disease, the use of renin-angiotensin system inhibitor, incretin related therapies, sodium-glucose cotransporter-2 inhibitor and statins. NRI and IDI were derived from logistic regression models (Model 3) with and without logarithm of baseline UACR to evaluate the clinical impact.The median and frequency of potential confounding variables were calculated. The relationships between PI and age, BMI, and other variables at baseline were estimated using Spearman’s rank correlation analyses. Owing to the relationship between baseline renal function and development of renal events, we also conducted analyses in a subgroup of patients with eGFR ≥ 60 ml/min/1.73 mP-value <0.05 was considered as statistically significant. The size, direction, and statistical significance of relationships were estimated by the odds ratio with 95% confidence interval (CI).All continuous variables are presented as median (interquartile range) or absolute number.


Small hepatocellular carcinoma (sHCC) is a special subtype of HCC with the maximum tumor diameter ≤ 3 cm and excellent long-term outcomes. Surgical resection or radiofrequency ablation provides the greatest chance for cure; however, many patients still undergo tumor recurrence after primary treatment. To date, there is no clinical applicable method to assess biological aggressiveness in solitary sHCC.In the current study, we retrospectively evaluated tumor necrosis of 335 patients with solitary sHCC treated with hepatectomy between December 1998 and 2010 from Sun Yat-sen University Cancer Center.P = 0.026, 0.003, respectively). The presence of tumor necrosis was associated closely with poorer cancer-specific overall survival (OS) and recurrence-free survival (RFS) as evidenced by univariate  and multivariate analysis . Notably, the combined model by tumor necrosis, vascular invasion and tumor size can significantly stratify the risk for RFS and OS and improve the ability to discriminate sHCC patients’ outcomes (P <  0.0001 for both).The presence of tumor necrosis was observed in 157 of 335 (46.9%) sHCC patients. Further correlation analysis showed that tumor necrosis was significantly correlated with tumor size and vascular invasion (Our results provide evidence that tumor necrosis has the potential to be a parameter for cancer aggressiveness in solitary sHCC. The combined prognostic model may be a useful tool to identify solitary sHCC patients with worse outcomes. As the second leading cause of cancer mortality worldwide, hepatocellular carcinoma (HCC) is an aggressive malignancy with its global incidence especially in Southeast Asia and Africa, with an increasing incidence in Europe and America –3. BecauSolitary sHCC is a special type of HCC with favorable long-term outcomes , 8.. AlbTraditionally, pTNM stage and histological grading systems are recognized as the most useful prognostic factors of sHCC. In addition, other features such as tumor size and vascular invasion are also utilized in clinical setting and found to be prognostic assessment of sHCC patients. Tumor necrosis is a common pathological feature of solid tumors, which is documented to be correlated with chronic ischemic injury due to the rapid growth of tumor –15. The The data of the pathologically proven and non-distant-metastasis solitary sHCC patients (335 cases) in Sun Yat-sen University Cancer Center  were retrospectively collected between December 1998 and 2009. In the current study, the selected patients were undergoing surgical resection (not ablation or transplantation) as the first therapy course. The eligibility criteria including: (1) solitary sHCC with the diameter ≤ 3 cm; (2) hepatitis B surface antigen positive; (3) primary and curative hepatectomy with a resection margin > 1 cm; (4) absence of metastasis and residual disease; (5) without preoperative adjuvant therapy; and (6) having complete follow-up information.The clinicopathologic information including patient age, gender, alfa-fetoprotein (AFP) level, alanine aminotransferase (ALT) level, tumor size, tumor capsule, histological differentiation, liver cirrhosis, vascular invasion and necrosis were collected, which are shown in Table Original histopathologic slides of the patients were previously reviewed by a senior pathologist (P Li), then independently re-confirmed by an experienced pathologist (M-Y Cai), both pathologists were blinded to the clinicopathologic prognostic data and re-examined simultaneously the slides to solve the discrepancies with a double-headed microscope. All of the selected patients had at least 3 tissue blocks, with a mean of 4.2  paraffin-embedded tissue blocks per tumor available for evaluation.The presence of tumor necrosis was carefully identified on hematoxylin-eosin (H&E)-stained slides, that characterized by homogenous clusters of sheets of dead cells, or coalescing groups of cells forming a coagulum, containing nuclear and cytoplasmic debris as previously described . CoagulaPatients were followed every 3 months by serum AFP level and ultrasound or computed tomography or magnetic resonance imaging at least every 6 months for more than 2 years after partial hepatectomy. All the enrolled patients were followed-up until January 18th, 2014. Re-resection when possible or transcatheter arterial chemoembolization, percutaneous ethanol injection or radiofrequency ablation was conducted in patients who had tumor recurrence. The definition of cancer-specific overall survival (OS) was the number of months from the date of surgery to the date of the last follow-up visit or time of death owing to sHCC. The definition of recurrence-free survival (RFS) was the number of months from the date of surgery to the first confirmability of cancer recurrence.P <  0.05 from a two-tailed test was considered statistically significant.Statistical analysis was computed using SPSS 19.0 software package . The χ2-test was used to evaluate the correlation between tumor necrosis and the clinicopathologic parameters of the sHCC patients. For univariate analysis, Survival curves were obtained by the Kaplan-Meier method, and the differences between groups in survival were performed by the Log-rank test. Multivariate survival analyses were conducted with the Cox proportional hazard regression model. A difference of Our current study identified 335 adult sHCC patients with long-term carriers of HBV and curative surgical resection. Some patients were followed by second-line treatments at the time of recurrence. Clinicopathological characteristics of the patients are detailed in Table Of the all patients, there were 295 (88.1%) males and 40 (11.9%) females, with a median age of 48 years. 196 (58.5%) patients had serum AFP level >  20 ng/ml among the cases with pre-operation serum AFP level record. 143 (42.7%) patients had serum ALT level >  40 μ/l. The median size of the tumors was 2.5 cm. Well-differentiated or moderate-differentiated tumors were identified in a total of 264 (78.8%) patients. 214 (63.9%) tumors were encapsulated. Vascular invasion was observed in 80 (23.9%) patients. Liver cirrhosis was presented in 134 (40%) patients.P = 0.026, 0.003, respectively; Table Presence of tumor necrosis was identified in 157 of 335 (46.9%) sHCC cases  and vascular invasion .Some clinicopathological parameters indicated a significant impact of prognosis, such as tumor size  and multivariate analysis . As we described previously [P < 0.001; hazard ratio, 2.663; 95% CI, 1.598–4.437) and tumor size was evaluated as an independent prognostic factor for patient survival .As variables examined to have prognostic influence by univariate analysis may covariate, the presence of tumor necrosis as well as other clinicopathologic features (tumor size and vascular invasion) was tested in multivariate analysis Table . The preeviously , of the P = 0.0200, 0.0020 and 0.0240, respectively). Meanwhile, tumor necrosis was associated with a decrease in OS and RFS of patients with AFP level ≤ 20 ng/ml and >  20 ng/ml  and RFS  in the current study based upon the combination of tumor necrosis, tumor size and vascular invasion.According to the results of the univariate and multivariate analyses, we conducted a new clinicopathologic prognostic model combining with three poor prognostic factors: tumor necrosis, tumor size and vascular invasion. Therefore, four subtypes were designated based on the presence of the three factors : subtype 1, absence of any risk factor; subtype 2, absence of any two risk factors; subtype 3, absence of any one risk factor; subtype 4, presence of three risk factors. This combined model could significantly stratify risk  for OS  might improve the ability to discriminate sHCC patients’ outcome, along with the effectiveness of personalized and precise treatment in the future.


Camellia vietnamensis is an economic woody plant producing high-value edible oils, which is commonly found and cultivated in south areas of China. To provide genetic information for future genetic research, we have sequenced and assembled the complete chloroplast (cp) genome of C. vietnamensis based on the Illumina Hiseq platform. The total genome size is 161,958 bp in length with 37% GC, which contains a large single copy  region, a small single copy  region, and a pair of inverted repeat  regions. It is comprised of 81 protein-coding genes, 44 transfer RNAs and 4 ribosomal RNAs. To obtain the phylogeny relationship, the cp genome of C. vietnamensis has been compared with other Camellia species; the results indicate that C. vietnamensis is closely related to C. taliensis. This study provides fundamental information of C. vietnamensis cp genome, and it is valuable to the molecular phylogenetic and genetic diversity analyses in future. Camellia vietnamensis is an economic woody plant, found mainly in Liuzhou city, Guangxi Province, China; it has now been widely used in producing edible oil from the seed kernels . The voucher specimen (CV_01) was reserved in State Key Laboratory of Tree Genetics and Breeding, Research Institute of Subtropical Forestry, Chinese Academy of Forestry.C. vietnamensis were collected from Liuzhou city . Total genomic DNA was extracted from fresh leaf tissue samples using MiniBEST plant Genomic DNA Extraction Kit . And the DNA concentration quality was controlled higher than 20 ng/µL  by NanoDrop 2000 . To perform sequencing, the Illumina libraries were constructed using TruSeq DNA sample Preparation kit , and 2 × 150 pair-end sequencing was carried out on the Illumina Hiseq 2500 platform  at Genesky Biotechnologies .The samples of Camellia japonica (NCBI accession Number: NC_036830.1) was selected as the reference genome sequence to filter the reads by Bowtie2   region of 86,657 bp, a small single-copy (SSC) region of 13,347 bp, and a pair of inverted repeats (IRs) of 30,977 bp. In total it contained 129 functional genes, comprised of 81 protein-coding genes, 44 transfer RNAs and 4 ribosomal RNAs.The assembled complete cp genome of Camellia species. The conserved protein sequences were extracted for alignment (Wang et al. C. vietnamensis was closely related to Camellia taliensis (C. vietnamensis would be utilised in molecular phylogenetic and genetic diversity analysis of camellias in the future.We performed a phylogenetic analysis using seventeen complete cp genomes of aliensis . The com


Serum levels of leptin and CRP were significantly reduced, while serum levels of adiponectin were elevated after NSPT, regardless of bodyweight or waist circumference (p < 0.05). Improvement in serum levels of leptin after NSPT was also observed in the Nwt with SP group (p = 0.015); these levels did not significantly differ in Nwt patients without SP. NSPT reduces serum levels of leptin and CRP and enhances serum levels of adiponectin in Thai patients with Owt/Ob, irrespective of periodontitis severity. These results suggest a role for periodontal treatment in the systemic inflammatory response of Thai people with Owt/Ob.Periodontitis and overweight or obesity independently change serum levels of leptin, adiponectin, and C-reactive protein (CRP). The aim of this study is to investigate the alterations of serum levels of leptin, adiponectin, and CRP after nonsurgical periodontal treatment (NSPT) in Thai patients with overweight or obesity (Owt/Ob) who did or did not exhibit severe periodontitis (SP) and normal weight (Nwt) patients with or without SP. Two hundred sixty patients were screened; 29 patients were included in this study. The study participants comprised 6 patients with Owt/Ob who exhibited SP, 11 patients with Owt/Ob who did not exhibit SP, 5 Nwt patients with SP, and 7 Nwt patients without SP. Periodontal disease status was evaluated; serum levels of leptin, adiponectin, and CRP were determined by enzyme-linked immunosorbent assay at baseline, as well as at 3 and 6 months after NSPT. At 3 months after NSPT, periodontal status was improved in all groups ( Duriunctions , 4. Furtin (CRP) . Therefoin (CRP) , 3.α, and interleukin-6 in patients with periodontitis; these are similar to the inflammatory mediators found in patients with Owt and Ob [In addition to its presence in people with Owt or Ob, chronic inflammation in the body may be found in patients with periodontitis, a major source of oral inflammation . Many prt and Ob , 6, 7.β, interleukin-6, and the tumor necrosis factor-α [An association between Ob and periodontitis was previously reported –10. Varifactor-α –20. Howefactor-α –18, 20. factor-α . A role TCTR20170607003). All participants received information regarding the study protocol and provided written consent to participate.This observational study was approved by the Institutional Review Board for human subjects of Mahidol University (COA. No. MU-DT/PY-IRB 2016/060.0411) and conducted in full accordance with the Declaration of Helsinki. The study was registered in the Thai Clinical Trials Registry on the WHO International Clinical Trial Registry Platform (registration number: 2). Patients were considered to have Owt/Ob if their BMI was ≥23 kg/m2 and WC was ≥85 cm for men or ≥80 cm for women; they were considered Nwt if their BMI was <23 kg/m2 and WC was <85 cm for men or <80 cm for women [Patients who attended the General and Special Clinic, Hospital for Tropical Diseases, Faculty of Tropical Medicine, Mahidol University, from November 2016 to December 2018 were recruited for this study. Inclusion criteria were as follows: Thai ethnicity, age ≥35 years, and possession of ≥9 teeth (excluding third molars and teeth indicated for extraction). Demographic data, medical histories, and dental histories of potential participants were collected by interview. Their levels of triglycerides, total cholesterol, and fasting blood sugar (FBS) within the preceding 6 months were evaluated from medical records. Potential participants were excluded if they met any of the following criteria: (1) presence of systemic diseases/conditions related to periodontitis and/or Ob, such as diabetes mellitus (FBS ≥ 126 mg/dl), hypertension, or cardiovascular disease; (2) presence of diseases that affect periodontal disease progression or treatment and/or gain/loss of weight, such as immunological disorders or osteoporosis; (3) current smoking habit or smoking habit within the past 1 year; (4) pregnancy or lactation; (5) the history of periodontal treatment during the preceding 6 months; (6) the history of antimicrobial, anti-inflammatory, immunosuppressive, and/or lipid-lowering therapies during the preceding 6 months; (7) regular use of mouth rinses containing antimicrobial drugs; and (8) use of orthodontic appliances. After assessment of the inclusion and exclusion criteria, the height, weight, and waist circumference (WC) were measured for the included patients. BMI was calculated as the weight in kilograms divided by the square of height  who had more than 30 years of experience and was unaware of each participant's BMI status. Periodontal measurements were taken at six sites per tooth , using a standard manual probe . The following clinical parameters were assessed: the percentage of sites with plaque , the percentage of sites with bleeding on probing (BOP), the probing depth (PD), and the distance from the gingival margin to the cementoenamel junction. In addition, the clinical attachment level  was calculated as the sum of PD plus the distance from the gingival margin to the cementoenamel junction at each site.The criteria proposed by the American Academy of Periodontology (with some modifications) were used in this study for diagnosis of periodontitis , 24. ParAfter baseline periodontal examination and blood sample collection, each participant was subjected to NSPT, which consisted of oral hygiene instruction, scaling, and root planing. A trained periodontist (NV) administered treatment. Oral hygiene instruction was given in accordance with individual needs to achieve a plaque score of <30% after treatment. Scaling and root planing under local anesthesia were performed on a quadrant-by-quadrant basis using a piezoelectric ultrasonic scaler , followed by Gracey curettes , until calculus could not be detected and the root surface felt smooth. Treatment (2–6 visits) was completed within 2 weeks. No pharmacological interventions were prescribed during treatment. The periodontal statuses of all participants were subsequently reassessed by the calibrated periodontist (PL), and oral hygiene care was provided at 3 and 6 months after NSPT by the trained periodontist (NV) .Serum levels of leptin, adiponectin, and CRP were evaluated at baseline, as well as at 3 and 6 months after NSPT. Two milliliters of serum from blood samples were separated by centrifugation and then aliquoted and stored at −80°C. Serum samples were thawed at room temperature, and cytokine measurement was performed by enzyme-linked immunosorbent assay using commercial kits for leptin, adiponectin, and CRP . Each serum sample was analyzed in duplicate. All assays were conducted in accordance with the manufacturer's instructions. The absorbance of each well was read using a microplate reader at 450 nm with a 650 nm reference wavelength. A blinded operator (ST) performed the assays.μg/ml of CRP between Owt/Ob and Nwt groups and assuming a standard deviation of 0.50 μg/ml, based on the findings of a previous cross-sectional study performed by our group [α of 0.05. Therefore, the number of participants for each of the four study groups was set at seven.The sample size to ensure adequate power for this study was calculated considering a difference of ≥0.55 ur group . FourteeU test (comparisons between two groups). The percentage distributions of participants with different patient profiles were compared using the chi-squared test. Values of BMI, WC, leptin, adiponectin, and CRP at baseline and after NSPT in each group were compared using the Friedman test, followed by the Wilcoxon matched-pairs signed-rank test. Changes in all four groups were compared between baseline and 3 months or between baseline and 6 months with respect to the plaque score, BOP, CAL, and PD; these comparisons were performed by means of the Kruskal–Wallis test. Regression analyses adjusted for the effects of age, sex, BMI, and WC were further analyzed to explore the associated factors with improvement of PD. The statistical significance threshold was set at p < 0.05 for all analyses.The data were analyzed with statistical software . The normality of the data distribution was assessed using the Shapiro–Wilk test. Cytokine values were normalized using logarithmic transformation. The mean age, FBS level cholesterol level, triglycerides level, CRP level, leptin level, adiponectin level, and periodontal status were compared among groups at baseline by using the Kruskal–Wallis test (comparisons among four groups) and Mann–Whitney Of the 260 participants screened, 229 were excluded because they did not meet the inclusion criteria; thus, 31 Thai individuals participated in this study. During the follow-up period, one patient in the Nwt with SP group developed tuberculosis, while one patient in the Owt/Ob with SP group was treated with anti-inflammatory drugs. Accordingly, these patients were excluded from the analysis. Therefore, the study participants consisted of 6 patients with Owt/Ob who exhibited SP, 11 patients with Owt/Ob who did not exhibit SP, 5 Nwt patients with SP, and 7 Nwt patients without SP .p = 0.01) and Nwt groups (p = 0.02). Although patients in the Owt/Ob group exhibited higher FBS and triglycerides levels than patients in the Nwt group , most blood chemistry values were at normal levels. Cholesterol levels also did not significantly differ among groups (p = 0.15) (Twenty-three (79.3%) patients were women. Patients in the Owt/Ob with SP group were significantly older than patients without SP, both in the Owt/Ob ( = 0.15) .p = 0.10 and p = 0.35, respectively). However, there was a significantly higher number of current alcohol consumers in the Owt/Ob group than in the Nwt group (p = 0.02). Regarding brushing habits, all patients brushed their teeth at least twice daily . However, BOP percentages in the Owt/Ob group did not significantly differ between patients with SP and patients without SP (p = 0.73). No statistically significant differences in plaque scores were found among participants in any of the groups (p = 0.21)  .p < 0.03). However, patients with SP tended to have higher leptin levels than patients without SP . Moreover, nine of 11 patients in the Owt/Ob without SP group had significantly greater WC (p = 0.027).There were no statistically significant differences in BMI and WC among groups after NSPT. Notably, five women in the Nwt without SP group exhibited significantly greater BMI and WC, compared with baseline (p < 0.05)  . Except  < 0.05) .p < 0.05 and p < 0.01, respectively). An opposite trend was observed in patients in the Owt/Ob without SP group at 6 months after NSPT (p < 0.05). Furthermore, CAL tended to decrease among patients in the Nwt without SP group at both 3 and 6 months after NSPT, but these differences were not statistically significant . Adjustment for the effects of age, sex, plaque score, BMI, and WC caused this difference in improvement of PD to disappear during regression analyses (data not shown).Among the four groups, changes between baseline and 3 months or baseline and 6 months did not significantly differ for plaque scores, percentage of BOP, or CAL, although the change in PD significantly differed among groups (st and 3rd quartile) serum levels of leptin, adiponectin, and CRP in all groups at baseline, as well as at 3 and 6 months after NSPT, are shown in The median . Compared with baseline, the mean leptin levels at 6 months after NSPT remained significantly lower among patients in the Owt/Ob group, irrespective of periodontitis status (p = 0.01 and p = 0.009). However, changes in leptin levels among patients in the Nwt without SP group were not significantly different at any timepoints after treatment . However, differences between 3 and 6 months were not statistically significant . At 6 months after NSPT, the mean CRP levels remained significantly lower than baseline among patients in the following three groups: Owt/Ob with SP, Owt/Ob without SP, and Nwt with SP. Nevertheless, no significant differences in CRP levels were observed between 3 and 6 months in any of the four groups  was lower than the threshold used in other studies [In this study, the threshold for Owt  or intermediate risk (1.0–3.0 μg/ml). Furthermore, levels of adiponectin, which is an anti-inflammatory mediator, were also elevated after NSPT in our study. These results confirm the systemic effects of NSPT on the systemic reduction of serum inflammatory mediators due to periodontal disease, irrespective of BMI or WC, and probably on decreased risk of coronary heart disease.Regarding the risks of cardiovascular disease, especially coronary heart disease, CRP levels >3  disease . Patient disease , 32. AltThis preliminary study had some limitations. The number of participants was limited due to the use of rigorous inclusion criteria, which were implemented to minimize the effects of confounding factors. Selection bias may have been involved because the included participants were patients who visited our hospital; there may also have been some biological variation among the patients. Therefore, these results should be carefully interpreted if they are generalized to other groups. More representative sample populations are needed in future investigations. Long-term longitudinal studies are needed to confirm the importance of NSPT in improvement of inflammatory mediators and reduction of the risk of cardiovascular disease. Despite these limitations, the findings of this study show that conservative periodontal treatment  can considerably reduce serum levels of leptin and CRP; it can also enhance serum levels of adiponectin during the 3–6-month recall period, especially in patients with Owt or Ob, irrespective of periodontitis severity.NSPT reduces serum levels of CRP and leptin while enhancing serum levels of adiponectin, in Thai patients with Owt or Ob, irrespective of periodontitis severity. These results support the importance of dental professionals in reduction of systemic inflammatory mediators in patients with a common noncommunicable disease.


Different mulches have variable effects on soil physicochemical characteristics, bacterial and fungal communities and ecosystem functions. However, the information about soil microbial diversity, community structure and ecosystem function in tea plantation under different mulching patterns was limited. In this study, we investigated bacterial and fungal communities of tea plantation soils under polyethylene film and peanut hull mulching using high-throughput 16S rRNA and ITS rDNA gene Illumina sequencing., Actinobacteria, Acidobacteria and Chloroflexi, and the dominant fungal phyla were Ascomycota, Mortierellomycota and Basidiomycota in all samples, but different mulching patterns affected the distribution of microbial communities. At the phylum level, the relative abundance of Nitrospirae in peanut hull mulching soils (3.24%) was significantly higher than that in polyethylene film mulching soils (1.21%) in bacterial communities, and the relative abundances of Mortierellomycota and Basidiomycota in peanut hull mulching soils  was significantly higher than that in polyethylene film mulching soils  in fungal communities. Peanut hull mulching increased the diversity of fungal communities in 0–20 cm soils and the diversity of bacterial communities in 20–40 cm soils. At the microbial functional level, there was an enrichment of bacterial functional features, including amino acid transport and metabolism and energy production and conversion, and there was an enrichment of fungal functional features, including undefined saprotrophs, plant pathogens and soils aprotrophs.The results showed that the dominant bacterial phyla were ProteobacteriaUnique distributions of bacterial and fungal communities were observed in soils under organic mulching. Thus, we believe that the organic mulching has a positive regulatory effect on the soil bacterial and fungal communities and ecosystem functions, and so, is more suitable for tea plantation. Mulching is an important measure in agricultural production, which is mainly used for soil improvement and environmental protection. According to the type of materials, it could be categorized as inorganic and organic mulching. Inorganic mulching was widely used as a low-cost and water-saving measure in agriculture areas that were susceptible to drought. Although it could improve soil moisture, prevent soil nutrient loss and control crop pests and diseases , 2, it cSoil microorganism played important roles in nutrient cycling and structure maintain in the agro-ecosystem , 9. SoilCamellia sinensis L.), an evergreen leafy plant, prefers acidic soil according to the genetic characteristics, thus, the soil environment of tea plantation is different from that of other crops. Several studies reported that mulching could increase the richness and diversity of microbial community in other crop soils [Tea plant  based on the bray–curtis distance. The first principal coordination axis accounted for 37.45 and 25.04% in bacterial and fungal communities, respectively. The second principal coordination axis explained 13.53 and 15.74% in bacterial and fungal communities, respectively. In bacterial communities, there were significant differences between 0 and 20 cm soils and 20–40 cm soils: CK2, F2 and P2 were on the positive side of the x-axis, and CK1, F1and P1 were on the negative side. Furthermore, there were significant differences between the mulching plastic film and peanut shell on the topsoil: P1 and P2 were on the positive side of the y-axis, and CK1, CK2, F1 and F2 were on the negative side  was significantly higher than that in F1 (1.21%) and CK1 (1.05%) of bacterial communities, and the relative abundances of Moretierella (33.66%) and solicoccozyma (10.60%) in P1 was significantly higher than that in F1 and CK1 of fungal communities.To dissect the composition of soil microbial community, we aligned the top30 OTUs with the Silva 119 and Unite database at the phylum and genus levels. The dominant bacterial phyla were Proteobacteria, Actinobacteria, Acidobacteria and Chloroflexi , Xanthobacteraceae (family), Parcubacteria (class), Amb_16S_1323 (the family and its genus), Candidatus_Magasanikbacteria (the order to genus), WS6_Dojkabacteria (the class to genus), UA_11 (the order to genus) and Paludibaculum (genus), and fungi included Piskurozymaceae (family and the genus Solicoccozyma) and Leucosporidiales (the order to genus). Three groups of bacteria and two groups of fungi in F1 were significantly enriched, of which, bacteria included Moraxellaceae (family), Solirubrobacteraceae (family) and Caulobacteraceae (family), and fungi included Acremonium (genus) and Emericellopsis (genus). The results showed that bacterial and fungal communities with significant differences in soils under different mulching were the noteworthy microbial communities in tea plantation.To identify the bacterial and fungal communities with significant differences in soils among three treatments, we further analyzed microbial community in 0-20 cm soil from phylum to genus level using LEfSe tool  database using PICRUSt Fig. a. The reIn addition, we assigned fungal OTUs to specific nutritional groups, and then subdivided them into specific ecological associations Fig. b. FunGuiThe mulching patterns on soils with different materials were widely adopted in agricultural cultivation for a long time. The general effect of mulching on soils has also been fully recognized. However, in terms of the influence of different mulching materials on the composition and function of microorganisms in soils, the depth of understanding is far from enough. In the study, in order to reveal the effects of mulching on microbial communities, we analyzed the bacterial and fungal diversity and community composition in tea plantation under polyethylene film or peanut hull using 16S rRNA and ITS rDNA, respectively. The results indicated that mulching could change soil properties and impact soil microbial diversity and community structure, and improve ecosystem functions of tea plantation.The diversity of soil microbial community was critical to the integrity, stability and sustainability of soil ecosystems . It was . Actinobacteria are mostly saprophytes in the soil. They were functionally diverse and contributed to the decomposition of organic matter, which they had ability to break down complex substrates to give them a competitive advantage over other bacteria [Sphingomonas and Nitrospira at on bacterial genus level . This region has atypical subtropical monsoon climate with an average annual precipitation of 1048 mm and an average temperature of 12.7 °C. The soil of the experimental field is brown soil, which is cleared and hoed manually before the experiment. The major soil parameters were as follows: pH 6.8, organic matter , available nitrogen , available phosphorus  and available potassium .2 under unified management and each treatment had three randomly replicate plots. The tea tree variety was ‘Zhongcha108’ with a row spacing of 1.5 m, and plant spacing of 0.33 m. The polyethylene film was black with a thickness of 0.004 mm, and was compacted with soil around the cover. The peanut hull was mulched in the soil surface only with a thickness of 10 cm. Prior to coring, mulch was removed from sampling area to prevent contamination of cores with surface organic matter. The soil sampling in every depth was homogenized by mixing together and passing through a 2-mm sieve for discarding mixed above-ground materials (plant residues and stones). Each soil sample with 3 replications was further divided into two parts: one part was frozen quickly in liquid nitrogen, and stored at − 80°Cuntil the microbial communities were analyzed; the other part was dried naturally in room and then analyzed for pH, available nitrogen (AN), available phosphorus (AP), available potassium (AK), organic matter (OM), total nitrogen (TN), total phosphorus (TP), total potassium (TK).The treatments were: (1) CK1: bare soil control, 0-20 cm soil; (2) CK2: bare soil control, 20-40 cm soil; (3) F1: mulching with polyethylene film, 0-20 cm soil; (4) F2: mulching with polyethylene film, 20-40 cm soil; (5) P1: mulching with peanut hull, 0-20 cm soil; and (6) P2: mulching with peanut hull, 20-40 cm soil. Each plot was 90 mAccording to the manufacturer’s protocols, microbial DNA from 18 samples was extracted from 18 samples in soils and the E.Z.N.A.® Soil DNA Kit  was used. The final DNA concentration and purified concentration and purification were determined by NanoDrop 2000 ultraviolet-visible spectrophotometer, and the DNA quality was checked by 1% agarose gel electrophoresis. The concentration and purification of final DNA were determined by NanoDrop 2000 UV-vis spectrophotometer , and the DNA quality was checked by 1% agarose gel electrophoresis. The DNA samples were individually amplified in V3-V4 hypervariable regions by PCR using primers 338F (5′-ACTCCTACGGGAGGCAGCAG − 3′) and 806R (5′-GGACTACHVGGGTWTCTAAT − 3′) for 16S rDNA in bacteria, and primers ITS1F (5′-CTTGGTCATTTAGAGGAAGTAA − 3′) and ITS2R (5′-GCTGCGTTCTTCATCGATGC-3′) for ITS in fungi. The procedure used in the PCR reaction had been described in previous study . In shorThe purified amplicons were pooled on the Illumina MiSeq platform  of equal molecular weight and paired-end sequencing (2 × 300) according to the standard protocol of MajorbioBio-Pharm Technology Co. Ltd. .http://drive5.com/uparse/) uses a novel “greedy” algorithm that could perform chimera filtering and OTU clustering at the same time. Using the confidence threshold of 70%, the classification of each 16S rRNA gene sequence was analyzed against the Silva database  through the RDP classifier algorithm (http://rdp.cme.msu.edu/). Using the confidence threshold of 70%, the classification of each ITS sequence was analyzed against the Unite database  through the RDP classifier algorithm.The original sequencing sequence was controlled using Trimmomatic software and merged by FLASH software. The specific criteria are consistent with previous study . UPARSE p < 0.05. Alpha diversity was applied to analyze microbial diversity, including the Chao1, Shannon, Ace, Simpson and rarefaction curve, which were calculated with Mothur software . The principal co-ordinates analysis (PCoA) was applied to reduce the dimension of the original variables based on the Bray-Curtis distances using R2.1.3. The Circos graph was built using Circos-0.67-7 software (http://circos.ca/) [http://huttenhower.sph.harvard.edu/galaxy/). The relationship between soil microbial community structures and soil environment factors was analyzed by Spearman correlation heatmap using pheatmap package.The means and standard deviations of the data were calculated and statistically examined by ANOVA and Duncan’s multiple range tests by SPSS . Significant differences were considered as cos.ca/) . Linear Additional file 1: Figure S1. The rarefaction curve of bacterial (A) and fungal (B) communities in soils under different mulching patterns.Additional file 2: Figure S2. The cladogram of the phylogenetic distribution of bacterial communities in soils under different mulching patterns.Additional file 3: Figure S3. The cladogram of the phylogenetic distribution of fungal communities in soils under different mulching patterns.


Ischemia-reperfusion injury of the spinal cord (SCII) often leads to unalterable neurological deficits, which may be associated with apoptosis induced by oxidative stress and inflammation. Astaxanthin (AST) is a strong antioxidant and anti-inflammatory agent with multitarget neuroprotective effects. This study aimed to investigate the potential therapeutic effects of AST for SCII and the molecular mechanism.Rat models of SCII with abdominal aortic occlusion for 40 min were carried out to investigate the effects of AST on the recovery of SCII. Tarlov’s scores were used to assess the neuronal function; HE and TUNEL staining were used to observe the pathological morphology of lesions. Neuron oxidative stress and inflammation were measured using commercial detection kits. Flow cytometry was conducted to assess the mitochondrial swelling degree. Besides, Western blot assay was used to detect the expression of PI3K/Akt/GSK-3β pathway-related proteins, as well as NOX2 and NLRP3 proteins.The results demonstrated that AST pretreatment promoted the hind limb motor function recovery and alleviated the pathological damage induced by SCII. Moreover, AST significantly enhanced the antioxidative stress response and attenuated mitochondrial swelling. However, AST pretreatment hardly inhibited the levels of proinflammatory cytokines after SCII. Most importantly, AST activated p-Akt and p-GSK-3β expression levels. Meanwhile, cotreatment with LY294002 (a PI3K inhibitor) was found to abolish the above protective effects observed with the AST pretreatment.Overall, these results suggest that AST pretreatment not only mitigates pathological tissue damage but also effectively improves neural functional recovery following SCII, primarily by alleviating oxidative stress but not inhibiting inflammation. A possible underlying molecular mechanism of AST may be mainly attributed to the activation of PI3K/Akt/GSK-3β pathway. Delayed postoperative paraplegia induced by spinal cord ischemia-reperfusion injury (SCII) is still one of the critical severe and catastrophic complications of thoracoabdominal aortic surgery. The incidence of paraplegia in patients undergoing that procedure is as high as 14% .Numerous studies have demonstrated that, following the primary mechanical injury to the spinal cord, oxidative stress occurs that destroys the normal balance of redox states and consequently contributes to the production of reactive oxygen species (ROS) , 4. Furtreduced isoflurane-induced neuroapoptosis via the phosphoinositide 3-kinases (PI3K) and their downstream target, protein kinase B (Akt) pathway, ultimately promoting axonal regeneration in the cerebral cortex and improving motor function [Astaxanthin (AST), a naturally occurring carotenoid pigment, is widespread in living organisms such as salmon, shrimp, microalgae, and yeast , 11. ASTfunction , which ifunction . In addifunction . Activatfunction . HoweverThus, we aimed to elucidate the role of AST pretreatment in SCII injury in rats and its molecular mechanism. The current study indicated that AST pretreatment attenuates SCII injury via activation of the oxidative stress-mediated PI3K/Akt/GSK-3β pathway.Male Sprague-Dawley rats  were obtained from the Experimental Animal Center of Hebei Medical University . They were housed in a standard light-controlled room (12:12-h light/dark cycle) and maintained at a constant temperature (25 ± 1 °C) and humidity (50–60%) for at least 1 week before the experiments. Food (GB 14924.3) and water were freely available. Experiments were carried out in accordance with the guidelines for the Care and Use of Laboratory Animals published by the US National Institutes of Health and approved by the Animal Care and Use Committee of the Second Hospital of Hebei Medical University. All efforts were made to reduce the number of animals used and minimize their suffering.AST  was resuspended in olive oil. All treatments were carried out between 08:30 and 10:30 a.m. to minimize the effects of circadian rhythm. MDA, XO, SOD, and GSH detection kits were obtained from Nanjing Jiancheng Bioengineering Institute .n = 10/group): (1) sham, in which animals underwent laminectomy surgery without an aortic occlusion clamp; (2) ischemia-reperfusion injury (SCII), in which rats underwent transient global spinal cord ischemia laminectomy with contusion lesion; (3) SCII+AST, in which rats received daily intragastric injections of 25 mg/kg AST for 14 days before SCII; and (4) LY+SCII+AST, in which rats received daily intragastric injections of 25 mg/kg AST and intravenous injections of LY294002  for 14 days before SCII [Forty rats were randomly assigned to four groups (ore SCII . All croore SCII . The ASTore SCII .SCII was induced in rats as previously described . In brien = 6/group) were anesthetized with 5% isoflurane and transcardiac perfusion with about 250 mL of cold normal saline. Next, the lumbar spinal cord (L2–5 segments) was quickly removed, and the samples were carefully dissected and then divided into two sections. One section was post-fixed using HE and TUNEL staining in order to observe cell survival and apoptosis. The other section was flash-frozen in liquid nitrogen and then stored at − 80 °C prior to further biochemical analyses, including the detection of oxidative stress and inflammatory cytokines using ELISA, as well as protein expression of NOX2, NLRP3, and PI3K/Akt/GSK-3β pathway using Western blot assay. Thereafter, the remaining 16 rats (n = 4/group) were decapitated, and flow cytometry was used to detect mitochondrial ROS and the degree of swelling.Rats were subjected to behavioral evaluation before surgery, 24 h, 48 h, and 72 h after surgery. After the last behavioral test was performed, 24 rats  level was determined by a method based on the reaction with thiobarbituric acid. Xanthine oxidase (XO) activity was determined spectrophotometrically at 293 nm based on the formation of uric acid from xanthine. Superoxide dismutase (SOD) activity was determined based on its ability to inhibit the oxidation of superoxide anions produced by a xanthine-xanthine oxidase system. One unit of SOD activity was defined as a 50% reduction of optical density at 450 nm. In addition, the GSH content of each group was measured at 412 nm. All procedures were performed under the guidance of the manufacturer’s instructions .The levels of IL-1β (RLB00), TNF-α (RTA00), and IL-10 (R1000)  as well as IL-18  were determined by ELISA kits according to the manufacturer’s instructions. They were expressed as picogram per milligram protein.n = 4/group) were sacrificed, and their lumbar spinal cords (L2–5) were quickly excised and homogenized. Subsequently, the homogenized samples were divided into two sections. One of the sections was immediately assessed by flow cytometry at 488 nm excitation and 530 nm emission. Data were analyzed using BDFACSAria™IICell Sorter software . The other section was used to detect the degree of mitochondrial swelling by measuring the decrease in optical density at 520 nm, as previously described [2PO4 and 3 mM sodium succinate (pH 7.2).Sixteen rats  based on our previously reported protocol . After dt test for multiple comparisons. P < 0.05 was considered as the significant difference.All data per group are expressed as the mean ± standard deviation (SD) and were analyzed using GraphPad Prism Progress, version 6.0 . The significance of differences between the groups was determined by one-way ANOVA followed by Tukey’s HSD post hoc To evaluate the effect of AST on functional recovery in SCII rats, hind limb motor function was recorded using the Tarlov scoring system Fig. . All ratTo observe the neuroprotective effect of AST on histopathology of the spinal cord in SCII rats, HE and TUNEL staining were performed. Notably, cells with Nissl-positive neuron were abundantly detected, and a centrally located nucleus was observed in the ventral horn of the spinal cord 72 h after injury in the sham group. However, compared with the sham group, the numbers of Nissl-positive neurons from the SCII group were lower and they displayed morphological deformation, including shrunken cell bodies and pyknotic nuclei  levels and antioxidative  activities in spinal cord homogenates were detected simultaneously. The levels of MDA and XO were higher in the SCII group. However, compared with the SCII group, the levels of MDA and XO in the SCII+AST group were remarkably lower, which means that AST strikingly mitigated the oxidative stress resulting from SCII-induced injury Fig. a, b.FigConcurrently, the activities of the antioxidative cytokines SOD and GSH were observed in all groups. As expected, compared with the sham group, there was a significant decrease in the levels of SOD and GSH in the SCII group. Interestingly, compared with the SCII group, the activities of SOD and GSH were higher in the SCII+AST group. However, this protective effect of AST was found to be counteracted by LY294002 cotreatment Fig. c, d.Compared with the sham group, the levels of proinflammatory cytokines  were higher and the level of the anti-inflammatory cytokine (IL-10) was lower in the SCII group Fig. . NeverthCompared to the sham group, mitochondrial ROS generation was significantly higher in the SCII group, whereas the mitochondrial ROS production was notably decreased in the SCII+AST group Fig. a. MeanwhTo further investigate the effects of AST on the molecular expressions of the oxidative stress cytokine (NOX2) and inflammatory index (NLRP3), Western blot analysis was performed. Strikingly, there were elevations of NOX2 protein and NLRP3 protein expressions in the SCII group when compared with the sham group Fig. . InteresTo investigate the underlying mechanism by which AST protects rats against SCII, we first detected the p-Akt and Akt levels using Western blotting assay; the change in the p-Akt/Akt ratio was slightly increased in the SCII group and was further increased in the SCII+AST group. Meanwhile, this upregulation was notably reversed by LY294002 cotreatment Fig. . Next, tIn the present study, we observed that AST pretreatment effectively improved motor functions and ameliorated pathological damages by reversing histological changes in the spinal cord. Furthermore, AST significantly inhibited oxidative stress and attenuated mitochondria swelling degree following the SCII. More importantly, PI3K/Akt/GSK-3β signaling pathway was found to be associated with the protective effects of AST on SCII-treated rats.The segmental blood supply of the spinal cord and relatively poor collateral circulation facilitate ischemia damage. SCII may further aggravate spinal cord damage. Various studies introduced a pathophysiological mechanism, associated with increases in oxidative stress and inflammatory processes with apoptosis, which plays a pivotal role in SCII , 22, 24.The cell membranes of spinal cord neurons are readily susceptible to oxygen free radical attacks. After SCII, the lipid peroxidation reaction was dramatically activated, while antioxidative activities strikingly decreased . TherefoAST, a dietary supplement with no significant adverse effects, is being investigated individually across a broad range of clinical applications, including cardiovascular health, acute pancreatitis, and neuropathic pain, all of which are associated with oxidative stress and inflammation in their pathogenesis , 16. BesGiven the numerous lines of evidence have implicated PI3K/Akt signaling pathway in modifying oxidative stress, GSK3β is a key active enzyme associated with downstream of Akt. Phosphorylation of GSK3β via Akt maintains this enzyme an inactive state and protects against tissue ischemic injury . Thus, wAs expected, we observed there were worse hind limb motor scores and that fewer neurons survived at 72 h after reperfusion in SCII rats compared to the sham group. Furthermore, oxidative products  and proinflammatory cytokines  were dramatically elevated in SCII rats, which is similar to our previous reports . InteresMoreover, we also evaluated NOX2 and NLRP3 protein expressions, which are important factors that prompt the expressions of many oxidant stress and inflammation , 37. Alt/GSK-3β. Similarly, these therapeutic effects were partially abolished when AST was combined with LY294002. This result might be explained that the nerve tissue had the capability of synthetizing and secreting some antioxidant enzymes to resist SCII-induced oxidative damage, although this capability did not satisfied the demand of cellular antioxidant defense [In order to further explore the possible molecular mechanism underlying the improved motor function of SCII rats pretreated with AST, we administered the PI3K inhibitor LY294002 as a comparative study and detected the levels of PI3K/Akt pathway-related proteins, including the ratios of p-Akt/Akt and p-GSK-3β defense . After aAST ameliorated SCII-induced oxidative stress and mitochondrial injury, as well as neuronal apoptosis in rats. Importantly, activation of PI3K/Akt/GSK-3β pathway-mediated antioxidative stress could play a critical role in the protective effects associated with AST pretreatment. Collectively, the data demonstrate that AST has the potential to serve as a therapeutic candidate for patients with SCII.


Liver transplantation induces self-injury and affects remote organs, such as the lung, kidney, and intestine. Postoperative intestinal dysfunction has been associated with prolonged hospitalization and affects a patient's health and quality of life. Electroacupuncture (EA) has been proven effective in multiple organ protection. However, the potential mechanism underlying the protective effects of EA on intestinal injury after liver transplantation remains unclear.  After establishing an autogenous orthotopic liver transplantation (AOLT) model, we studied the effects of EA pretreatment on intestinal injury after AOLT. We used the JAK2-specific inhibitor AG490 to explore the underlying mechanism. Histological analysis and apoptosis assays were used to evaluate intestinal injury. Oxidative stress index and inflammatory response were also measured after AOLT. Furthermore, we detected the phosphorylation levels of JAK2, STAT1, and STAT3 by Western blot.  We found that pretreatment with EA alleviated intestinal injury after AOLT, as shown by HE staining and TUNEL methods. EA pretreatment inhibited the expressions of p-JAK2, p-STAT1, and p-STAT3 in the intestines after AOLT. Upon treatment with JAK2-specific inhibitor AG490, intestinal injury was balanced.  The data indicated EA pretreatment alleviated intestinal injury after AOLT by inhibiting the JAK/STAT signaling pathway. These results provide basic evidence to support the potential therapeutic efficacy of EA. Liver transplantation is the most effective method to address end-stage liver diseases. During the past few decades, remote organ injury has been widely studied during liver transplantation. Liver transplantation not only induced self-injury but also affected remote organs, such as the lung, kidney, and intestine –4. With Acupuncture is a traditional Chinese medical treatment and is simple to perform, safe, and reliable. Electroacupuncture (EA) has been proven effective in multiple organ protection –8. A preThe Janus kinase/signal transducer and activator of the transcription (JAK/STAT) signaling pathway is involved in a wide range of distinct cellular processes, including inflammation, apoptosis, cell-cycle control, and development . The JAKA total of 40 adult male Sprague-Dawley rats (220-250 g) were purchased from the People's Liberation Army Military Academy of Medical Sciences Laboratory Animal Center. The animals were housed in 12 h light-dark cycles with controlled room temperature and were fed with regular rat chow and water ad libitum but were fasted 12 h before experiments. Animals were randomly assigned into five groups: group A, sham operated (sham group); group B, autogenous orthotropic liver transplantation (AOLT group); group C, pretreated with EA  for 3 days+AOLT (EA+AOLT group); group D, pretreated with EA for 3 days+sham operated (EA+sham group); and group E, pretreated with EA for 3 days+AG490  30 min before establishing the AOLT model (EA+AOLT+AG490 group). The dosage was determined from a previous study [2 and were prone on a heating blanket. Rats in the sham group underwent laparotomy without performing AOLT as control. Total liver ischemia was induced by clamping the hepatic artery, the portal vein, suprahepatic vena cava (SHVC), and intrahepatic vena cava (IHVC). After clamping, the portal vein and IHVC were cannulated with a polyethylene tube, and the liver was perfused through the portal vein with 500 mL of heparinized cold saline  (2.5 IU/mL) to wash out all blood from the liver. The tubes were removed after perfusion, and the openings of tube insertion were closed. The SHVC and IHVC were then declamped immediately after repairing the vessels. After 45 min, the clamps were removed to allow the return of blood flow to the liver. After 6 h, the animals were euthanized by administering CO2 inhalation. Blood samples and intestinal tissue biopsies were taken. The tissues were fresh frozen for histological and biochemical evaluation.  EA stimulation of the ST36 acupoint can alleviate the inflammatory response caused by Complete Freund's adjuvant (CFA) , 17. Zusμm thick sections, which were stained with hematoxylin and eosin (HE). The damage of intestinal mucosal cells was analyzed by the Chiu score method [Formalin-fixed intestinal tissue was dehydrated, embedded in paraffin, and sliced into 4 e method : 0—normaμm thick paraffin sections were deparaffinized, treated with 0.1% TritonX-100, and incubated at 37°C for 8 min in a humidified atmosphere. PBS wash was performed 3 times. Then, the sections were further incubated with a mixture of marker and enzyme solution at 37°C for 1 h. PBS wash was performed 3 times, and the samples were observed under a fluorescence microscope  at 450-550 nm. Five high-power fields were randomly selected to determine the percentage of apoptotic cells.To detect the apoptotic positive cells, terminal deoxynucleotidyl transferase-mediated digoxigenin-labeled UTP nick end labeling assay (TUNEL) was performed using an ApopTag peroxidase in situ cell death detection kit . Briefly, the 4 α (TNF-α), high-mobility group box-1 (HMGB-1), intestinal fatty acid-binding protein (IFABP), and endotoxin (LPS) were examined using an ELISA kit  according to the manufacturer's instructions [Blood samples were collected from all rats. The serum levels of tumor necrosis factor-ructions .W/D ratio was calculated as the ratio of the wet weight to the dry weight of the intestine.To test the edema of intestinal after AOLT 6 h following reperfusion, 5 cm of ileum tissue at 2 cm from the back of the blind was harvested, cleaned by removing blood and water, and weighed. Then, the sample was incubated at 70°C for 24 h and weighed. The μmol/g tissue. SOD activity was measured according to the Paoletti and Mocali method. In this assay, superoxide anion was generated from molecular oxygen in the presence of EDTA, manganese II chloride, and mercaptoethanol. Nicotinamide adenine dinucleotide phosphate oxidation was linked to the availability of superoxide anions in the medium.The levels of malondialdehyde (MDA) and superoxide dismutase (SOD) were determined using assay kits  according to the manufacturer's instructions . The tisμm thick sections of the fixed intestines were dewaxed with xylene, hydrated in graded concentrations of ethanol, and treated with 0.3% hydrogen peroxide for 10 min to quench endogenous peroxidase. The sections were heated in a microwave oven in sodium citrate (pH = 6.0) at 95-99°C for 15 min and then cooled at room temperature. The sections were incubated in 5% blocking serum for 30 min and then in primary antibodies  at 4°C overnight. They were subsequently incubated with biotinylated secondary antibodies for 30 min and finally counterstained with hematoxylin.Immunohistochemical staining of the intestinal tissue was performed on formalin-fixed paraffin sections using a microwave-based technique. The 4 The protein lysates were prepared from frozen tissues in ice-cold RIPA buffer (Sigma-Aldrich). The extracted protein was separated in a 10% sodium dodecyl sulfate- (SDS-) PAGE and then electrophoretically transferred to a nitrocellulose membrane . Membranes were blocked with 5% nonfat milk powder in TBS for 1 h at room temperature. These membranes were subjected to immunoblot analysis with antibodies to JAK2, p-JAK2, STAT1, p-STAT1, STAT3, and p-STAT3. The protein antibody immune complexes were detected with horseradish peroxidase-conjugated secondary antibodies and enhanced chemiluminescence reagents . GAPDH was used as an internal control to calculate the ratio of optical density, and values were compared with those of sham controls.P < 0.05 was considered significant.All values are given as the mean ± SD. Statistical analysis was carried out using GraphPad Prism 8.0 software . One-way analysis of variance was used to compare the measurement data between groups. To examine the protective effect of EA pretreatment against intestinal injury after AOLT, each rat was analyzed by histopathology, and the intestinal mucous damage score was calculated. A normal histological structure was observed in the sham group by HE staining. Severe disruption of structural integrity in brush border, including loss of mucus, villi, and widespread necrotic area, was observed in the AOLT group. However, these damages were improved in the EA pretreatment group. The intestinal mucous damage score significantly increased in the AOLT group compared with the sham group. EA pretreatment combined with AG490 improved intestinal damage; no significant differences were observed between the EA+sham group and the EA+AOLT+AG490 group Figures .W/D ratio changes in all groups. As shown in W/D ratio caused by AOLT. When compared with the AOLT group, the intestinal W/D ratio decreased in the EA+sham and EA+AOLT+AG490 groups. The EA+sham group did not show a significant difference from the sham group . However, EA application decreased the expression of p-JAK2, p-STAT1, and p-STAT3 proteins (P < 0.05 vs. the AOLT group). Treatment with AG490 reduced the expressions of p-JAK2, p-STAT1, and p-STAT3 proteins (P < 0.05 vs. the AOLT group). These results demonstrated that EA inhibited the JAK/STAT signaling pathway in intestinal injury after AOLT. The JAK/STAT signaling pathway was active in the AOLT group . EA application was associated with the occurrence of apoptosis of epithelial cells, which was less than that observed with the AOLT group (P < 0.05). In the EA+sham group, EA treatment had no effect on apoptotic epithelial cells, comparable with the sham group. The EA+AOLT+AG490 group had similar results as the sham group  and oxidative stress factors (MDA) increased significantly in the AOLT group. However, in the EA+AOLT group, apoptosis of epithelial cells resulting from the intestinal injury was significantly decreased. Caspase-3, which participates in the apoptosis pathway in the intestinal mucosa, was also markedly reduced in the EA+AOLT group, which confirmed the decreased disruption of the structural integrity of intestinal mucosa, as observed by HE staining. This study evaluated the protective and antiapoptotic effects of EA on intestinal architecture and apoptosis in intestinal injury.Numerous strategies have been designed to reduce intestinal injury after AOLT . AlthougJAK/STAT is involved in many pathophysiological processes in the body , 46. JAKTaken together, intestinal injury contributed to the damage of intestinal function and histological structure and enhanced the apoptosis of intestinal epithelial cells and the expression of protein Cleaved Caspase-3. We demonstrated that EA pretreatment attenuated intestinal injury by inhibiting the JAK/STAT signaling pathway in a rat AOLT model. Zusanli is a classic and preferred acupoint for treating gastrointestinal diseases and is widely used in clinical practice. Its therapeutic effect is recognized by many medical workers and patients. EA is a branch of acupuncture and moxibustion therapy. Compared with classic acupuncture, it has many advantages, such as good therapeutic effects, a wide range, ease of stimulation control, and continuous needle movements . HoweverIn conclusion, our study showed that EA reduced intestinal injury against AOLT, at least in part through its inhibitory effects on the injury-induced activation of the JAK/STAT signaling pathway. If extrapolated to a clinical setting, EA has the potential to serve as a clinical strategy for preventing perioperative intestinal injury after AOLT.


Microphysogobio kiatingensis is endemic freshwater fish of China and mainly distributed in the middle and upper reaches of the Yangtze River. The complete mitogenome of M. kiatingensis was first determined by using the next-generation sequencing in present study. The circular mitogenome followed the expected pattern for vertebrates, being 16,603 bp in length. It contained 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, one displacement loop (D-loop) locus and an origin of replication on the light-strand (OL). Most of the genes are encoded on the heavy strand except ND6 and eight tRNA genes. The overall base composition was 30.78% A, 26.09% T, 26.59% C, 16.54% G, with 56.87% AT. Thirteen intergenic spacers and nine gene overlaps exist in the complete mitochondrial genome. Phylogenetic analysis based on the tandem 13 coding protein genes nucleotide sequences with two different methods  indicated that M. kiatingensis was grouped in one clade with the same genus. The complete mitogenome of M. kiatingensis can provide a useful data for the further research in Gobioninae. Meanwhile, it also provides help for biological genetics and classification of species. Microphysogobio kiatingensis , originated in the Leshan, Sichuan, is endemic freshwater fish of China and mainly distributed in the middle and upper reaches of the Yangtze River  in this study. The specimens were collected from Chengdu, Sichuan province of China  in September 2017 and were stored in Zoological Specimen Museum of Neijiang Normal University (accession number: 20170925BB05). A 30–40 mg fin clip was collected and preserved in 95% ethanol at 4 °C. Total genomic DNA was extracted from these caudal fins by a Tissue DNA Kit  following the manufacturer’s protocol. Subsequently, the genomic DNA was sequenced using the NGS, and then the mitogenome was assembled using Abbottina rivularis as reference.The complete mitochondrial DNA sequence of Lys) to 76 bp (tRNALeu). Moreover, 12S rRNA (958 bp) and 16S rRNA (1691 bp) were located between tRNAPhe and tRNALeu and separated by the tRNAVal. Additionally, two non-coding regions were D-loop (927 bp) and OL (31 bp), which may control mitogenome’s replication and transcription.The circular mitogenome followed the expected pattern for vertebrates, being 16,603 bp in length (GenBank Accession number MG797640). It contained 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, one displacement loop (D-loop) locus and an origin of replication on the light-strand (OL). In these genes, ND6 and eight tRNA genes were encoded on light strand while the rest of genes were encoded on heavy strand and the arrangement of these genes were the same as that found in teleosts  and maximum likelihood (ML) methods using the 13 PCGs  was defined as an outgroup. The 13 taxa all belong to Cyprinidae, Gobioninae except C. argus. The other 12 species from three genus  were divided into three clades (Microphysogobio has a closer relationship with Abbottina than Sarcocheilichthys. These results may provide an important information for morphological classification of Gobioninae species and the study of biological origin.So far, the mitochondrial PCGs have been widely used for inferring phylogenetic relationships (Boore et al. e clades . The sam


Schistosoma and gut bacteria in mice and humans, no data is yet available on whether worm-mediated changes in microbiota composition are dependent on the baseline gut microbial profile of the vertebrate host. In addition, the impact of such changes on the susceptibility to, and pathophysiology of, schistosomiasis remains largely unexplored. In this study, mice colonized with gut microbial populations from a human donor (HMA mice), as well as microbiota-wild type (WT) animals, were infected with Schistosoma mansoni, and alterations of their gut microbial profiles at 50 days post-infection were compared to those occurring in uninfected HMA and WT rodents, respectively. Significantly higher worm and egg burdens, together with increased specific antibody responses to parasite antigens, were observed in HMA compared to WT mice. These differences were associated to extensive dissimilarities between the gut microbial profiles of each HMA and WT groups of mice at baseline; in particular, the gut microbiota of HMA animals was characterized by low microbial alpha diversity and expanded Proteobacteria, as well as by the absence of putative immunomodulatory bacteria (e.g. Lactobacillus). Furthermore, differences in infection-associated changes in gut microbiota composition were observed between HMA and WT mice. Altogether, our findings support the hypothesis that susceptibility to S.mansoni infection in mice is partially dependent on the composition of the host baseline microbiota. Moreover, this study highlights the applicability of HMA mouse models to address key biological questions on host-parasite-microbiota relationships in human helminthiases.In spite of growing evidence supporting the occurrence of complex interactions between  More th species . In addichildren . Over thh humans –18 and eh humans , 19–21.   following experimental infections with S. mansoni cercariae, and compared the findings with those observed in wild type (WT) animals. Strikingly, higher worm and egg burdens were observed in HMA mice compared to WT animals, and these findings were associated with specific changes in the microbiota composition of each rodent line.While the causality of somiasis , 19, 21.S. mansoni (NMRI strain) is maintained at the Wellcome Sanger Institute (WSI) by breeding and infecting susceptible snails  and mice. All mouse infections and regulated procedures described in this study were presented to and approved by the Animal Welfare and Ethical Review Body (AWERB) of the WSI. All experiments were conducted under the Home Office Project Licenses (Procedure Project License - PPL) No. P77E8A062 held by GR, and No. P6D3B94CC held by TL. The AWERB is constituted as required by the UK Animals (Scientific Procedures) Act 1986 Amendment Regulations 2012.The life cycle of 2, 10% H2, 10% N2). Five male and five female C57BL/6 germ-free mice, bred at the WSI, were inoculated by oral gavage with 200 μl of donor homogenates once a week, for 3 weeks. One week after the final gavage, mice were set up as breeding pairs in a decontaminated positive pressure isolator. Mice used in this study belonged to the 4th generation of breeding animals. All consumables that entered the isolator for maintenance of the colony were autoclaved at 121°C for 15 min. Feces from parental HMA mice (i.e. germ-free transplanted with human feces) were analyzed by 16S rRNA gene amplicon sequencing, in order to confirm engraftment of the human flora (unpublished data). Animals used for experiments were removed from the isolator in sealed ISOcages and maintained on a positive pressure ISOrack (Tecniplast).The HMA mouse line was generated at the WSI. Briefly, fresh feces from a healthy human donor, processed within 1 h from transportation to the laboratory, were homogenized at 100 mg/ml in 1x D-PBS in an anaerobic cabinet (80% COS. mansoni cercariae (Sm-exp), as described previously . Experimental infections, with matched uninfected controls, were performed in two independent batches (B1 and B2), as indicated in A total of 10 HMA and 10 WT  mice were exposed to 80 eviously , and maiSm-exp mice were euthanized, S. mansoni adult worms were recovered by portal perfusion, and livers removed for parasite egg counts (Sm+) WT and HMA mice were assessed by unpaired two-tailed t-test, following the application of the Kolmogorov-Smirnov normality test for assessment of normal distribution of worm and egg counts datasets; differences in female fecundity were evaluated by Mann-Whitney test. Control Sm- mice were perfused under the same conditions as Sm-exp animals and the perfusates were discarded. For microbiota profiling, fecal pellets were collected directly from the colons of Sm-exp and Sm- mice of each line (i.e. WT and HMA) at necropsy. Samples were transferred to sterile tubes, snap frozen in dry ice, and stored at −80°C until DNA isolation, which was performed within 3 weeks from sample collection.Serum samples were obtained from blood collected by puncture of the tail vein at days 28, 42, and 49 post-cercarial exposure (p.i.). at days 28, 42, and 49 post-infection. At day 50 p.i., g counts . BrieflySm+ and three Sm- animals from each HMA and WT groups were randomly selected to investigate the kinetics of S. mansoni-specific antibodies. Endotoxin-free soluble egg antigen (SEA) was generated from S. mansoni eggs isolated from trypsinized livers of infected mice, as previously described , six WT and six HMA Sm- mice, as well as four negative controls were processed for high-throughput sequencing of the bacterial 16S rRNA gene (® NEBNext Hot Start High-Fidelity DNA Polymerase (New England Biolabs), 5 ng/μl of template DNA and the following thermocycling protocol: 98°C for 2 min, 20 cycles of 98°C/15 s - 63°C/30 s - 72°C/30 s, and a final elongation step at 72°C for 5 min. Amplicons were purified using AMPure XP beads (Beckman Coulter) and set up for the index PCR using Q5® NEBNext Hot Start High-Fidelity DNA Polymerase (New England Biolabs) and Nextera XT index primers (Illumina) with thermocycling conditions as follows: 3 min at 95°C, eight cycles of 30 s at 95°C – 30 s at 55°C – 30 s at 72°C, and 5 min at 72°C. The indexed samples were purified using AMPure XP beads, quantified using Qubit™ dsDNA High Sensitivity Kit (Life Technologies) and equal DNA amounts from each sample were pooled. The resulting pooled library was quantified using the NEBNext® Library Quantification Kit for Illumina® (New England Biolabs), and sequenced using the v3 chemistry . DNA isolation and sequencing were performed separately for samples collected from each B1 and B2 mice  negative controls, using the PowerSoil DNA Isolation Kit (Qiagen) according to manufacturers’ instructions. A total of 28 samples including eight WT and eight HMA RNA gene Table 1.RNA gene  containiCHELINdb .https://qiime2.org) software suite for downstream analysis   , and thegenome.net/calypso/)  on Bray-Curtis dissimilarities and supervised Canonical Correlation Analysis (CCA) were performed using ‘mouse line’ (i.e. WT and HMA) and ‘infection status’ (i.e. Sm- and Sm+) as explanatory variables, respectively. Beta diversity was calculated using Bray-Curtis dissimilarity and differences between groups were assessed using Analysis of Similarity (ANOSIM) (via multiple linear regressions by incorporating worm burdens (or EPG of liver) as the dependent variable and bacterial genera as explanatory variables. Fecal microbial alpha diversity (Shannon index), richness and evenness were calculated using the ‘diversity’ and ‘specnumber’ functions of the vegan R package and differences between groups were evaluated by Mann-Whitney test.Statistical analyses were performed using the Calypso software  . Relativ(ANOSIM) . Differe(ANOSIM) . CorrelaSm+ or Sm- mice in each line (i.e. WT and HMA) were identified by prediction of correlation networks, with microbial genera represented as nodes, genera abundances as node size, and edges representing positive associations among them. Networks were generated by computing associations among microbial taxa using Pearson’s correlation. The resulting pairwise correlations were converted into dissimilarities and used to ordinate the nodes in a two-dimensional plot by PCoA; thus, correlating nodes were located in close proximity, while anti-correlating nodes were placed at distant locations in the network. Taxa abundances were associated with the type of infection using Pearson’s correlation and the nodes were colored based on the strength of this association.Clusters of co-occurring bacteria and their association with S. mansoni, whereas no adult worms or eggs were recovered from two Sm-exp WT animals included in the second experiment at post mortem; the latter were thus removed from subsequent analyses  and (ii) exposure to environmental microbes (maintained under specific-pathogen free [SPF] conditions vs. in isolation). We anticipated that these profound dissimilarities might significantly impact the composition of the gut microbiota of each mouse line; therefore, we first investigated structural and compositional differences between the gut microbial communities of these rodent lines at baseline. Fecal microbial profiles obtained from samples from Sm- WT and HMA were clustered separately by PCoA, which led to the identification of a putative outlier sample in the former group , and thus sequence data obtained from this sample were removed prior to further analyses (The two rodent lines employed in this study were characterized by marked differences in (i) the origin of the microbiome , followed by the genera Bacteroides (10.23 ± 3.62%), Prevotella UCG-001 (5.11 ± 4.84%) and Lactobacillus (4.28 ± 3.50%). In contrast, the genus Bacteroides was predominant in the gut of HMA mice (38.31 ± 3.69%), followed by Parabacteroides (10.82 ± 0.94%), Lachnospiraceae NK4A136 group (8.19 ± 4.89%) and Parasutterella (6.12 ± 1.75%)  and Lactobacillus (4.28% ± 3.50%) were exclusively detected in WT, while Barnesiella (3.00 ± 1.10%) in HMA mice  was associated with a significant decrease in evenness , whereas that observed in Sm+ HMA mice  was linked to a significant decrease in ASV richness   and S. mansoni infection in WT mice , as well as the genus Butyricimonas  ; the latter bacterial class (and related taxa) was solely detected in the gut microbiota of HMA mice  in Sm+ WT hosts  were investigated by multiple regression. While the genera S. mansoni infection in HMA and WT mice revealed substantial differences between rodent lines, some common traits were identified. In order to identify fecal microbial taxa consistently associated with infection in both mouse lines, the LEfSe workflow was applied to all metagenomics datasets, while controlling for both infection status and rodent line. In accordance with the findings obtained from individual analyses of each rodent line, the genus Eubacterium coprostanoligenes group was positively associated to S. mansoni infection when the two variables (i.e. ‘infection status’ and ‘mouse line’) were considered, whereas the genera Ruminococcus 1, Butyricicoccus and Family XIII UCG-001, among others, were associated to Sm- samples  have been associated with S. haematobium infection in adolescents from an endemic region  at the expense of others, generally resulting in intestinal dysbiosis , which is partially consistent with data from studies performed in humans from endemic areas, where no alterations in alpha diversity had been reported  in the mouse gut have been linked to differences in immune responses and susceptibility to infection by the gastrointestinal helminth Heligmosomoides polygyrus (Lactobacillaceae (including the genus Lactobacillus) were only detected in the gut of WT mice, although populations of these bacteria were not altered by infection. Previously, transient expansions of Lactobacillus were reported in the gut of Swiss-Webster mice exposed to large numbers of S. mansoni cercariae  of this genus of bacteria in modulating the response against S. mansoni in HMA mice merit(s) further investigations.The genus e models –47. In cescribed . IFN-γ h studies –51. More studies . TherefoRuminococcus 1 (the abundance of which was negatively correlated with infection burdens in both WT and HMA mice) includes mucin-degrading bacteria  (Butyricicoccus (Shuttleworthia (Intestinimonas (S. mansoni infection burdens (the latter two in HMA mice only); thus, reductions in the abundances of these populations of gut bacteria might lead to an impaired regulation of intestinal immunity and barrier function in infected animals  , 61. Othcicoccus , and theeworthia  and Inteinimonas , 65, wer animals .Sm+ WT and HMA mice, butyrate-producing Roseburia  and/or selected human microbes exerts on S. mansoni infection rates. In particular, such studies should include (i) experimental infections of additional HMA mouse lines, (ii) groups of control rodents derived from germ-free mice transplanted with the fecal microbiota of WT mice, and (iii) antibiotic-treated WT mice. Moreover, predictive models linking infection burdens to the abundances of key gut microbes and/or alpha diversity prior to infection could be useful to identify gut microbial features that might facilitate parasite colonization. This knowledge will underpin future mechanistic studies to untangle the complex interactions between the gut microbiome, the host immune response and the outcome of schistosome infection.Overall, our results suggest that the baseline composition of the host gut microbiome might impact host susceptibility to The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/The animal study was reviewed and approved by Animal Welfare and Ethical Review Body (AWERB).ACor, GR and CC designed the research. ACor, SC, ACos, CM, KH, CB, CT, and GR carried out the research. ACor, ACos, AA and JR performed data processing and analyses. ACor, GR, and CC wrote the main manuscript text with input from AM, TL, and MB. The figures were prepared by ACor and ACos. All authors reviewed the manuscript. All authors contributed to the article and approved the submitted version.ACor is supported by Red de Investigación Colaborativa en Enfermedades Tropicales—RICET . Wellcome provided core-funding support to the Wellcome Sanger Institute, award number 206194. JR is the grateful recipient of a PhD scholarship by the UK Biotechnology and Biological Sciences Research Council (BBSRC). Research in the CC laboratory is funded by grants by the Isaac Newton Trust and the University of Cambridge.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.