Humans ; Gastrointestinal Microbiome ; Constipation/therapy* ; Probiotics/therapeutic use* ; Patients

Objective To explore the effect of quercetin combined with iron death inhibitor Ferrostain-1 on oxalate-induced HK-2 cell injury.Methods HK-2 cells were randomly divided into control group(normal cultured cells),model group(0.5 mmol·L-1 calcium oxalate crystals),quercetin group(0.5 mmol·L-1 calcium oxalate crystals+100 μmol·L-1 quercetin),inhibitor group(0.5 mmol·L-1 calcium oxalate crystals+8 μmol·L-1 Ferrostain-1)and combination group(0.5 mmol·L-1calcium oxalate crystals+100 μmol·L-1quercetin+8 μmol·L-1 Ferrostain-1).Cell counting kit-8(CCK-8)assay was used to detect cell survival rate;Western blot was used to detect iron death related protein expression such as glutathione peroxidase 4(GPX4);flow cytometry and Tunel assay were used to detect cell apoptosis,and assay kit was used to detect cellular iron ions and antioxidant levels.Results The cell survival rates of control group,model group,quercetin group,inhibitor group and combination group were(100.00±2.55)％,(54.49±4.11)％,(64.26±6.30)％,(58.03±3.04)％and(79.37±4.29)％,respectively;GPX4 protein expression levels were 0.98±0.11,0.33±0.05,0.56±0.05,0.78±0.07 and 1.11±0.11,respectively;cell apoptosis rates were(4.15±0.28)％,(23.12±2.49)％,(17.28±1.07)％,(15.08±1.41)％and(8.95±0.75)％,respectively;Fe2+levels were(100.00±0.87)％,(162.55±14.70)％,(149.09±9.50)％,(144.95±11.12)％and(131.76±12.18)％,respectively;superoxide dismutase(SOD)levels were(58.67±3.46),(21.56±1.32),(33.60±2.03),(35.15±3.02)and(44.27±3.89)U·mL-1,respectively.The above indicators of the model group were compared with the control group,and the above indicators of the quercetin group,inhibitor group,and combination group were compared with the model group,the above indicators of the combination group were compared with the quercetin group,and inhibitor group,all they all showed statistical significance(all P＜0.05).Conclusion Iron death inhibitors can enhance the inhibitory effect of quercetin in vitro on oxalate-induced renal tubular epithelial cell injury.

AIM To investigate the renoprotective effects and mechanism of Caragana sinica roots,Astragali Radix and their combination use on the rat model of diabetic kidney disease(DKD).METHODS Sixty SD rats were randomly divided into the normal group,the model group,the Empagliflozin group(10 mg/kg),the C.sinica roots group(3.1 g/kg),the Astragali Radix group(3.1 g/kg),and the C.sinica roots plus Astragali Radix group(6.2 g/kg).In contrast to the intact rats of the normal group,rats of the other groups underwent left nephrectomy and intraperitoneal injection of streptozotocin(STZ)followed by 8-week intragastric gavage of the corresponding agent,during which their levels of FBG and 24 h urinary microprotein(24 h U-mAlb)were detected regularly.The rats killed at the end of the trial had their levels of Scr,BUN and Cystatin C detected;their renal pathological changes observed by HE,PAS and Masson stainings;their expressions of macrophage marker proteins CD68 and iNOS detected by immunohistochemistry;their expressions of renal JNK/SAPK pathway proteins such as JNK,p-JNK,TNF-α,IL-1β and ICAM-1 detected by Western blot;and their serum levels of TNF-α,IL-1β and ICAM-1 detected by ELISA as well.RESULTS Compared with the normal group,the model group displayed increased levels of FBG,24 h U-mAlb,BUN,Scr and Cystatin C(P＜0.01);more renal pathological damage,and increased levels of TNF-α,IL-1β and ICAM-1 in the renal tissue and serum(P＜0.01);and increased renal protein expressions of JNK and p-JNK(P＜0.01).Compared with the model group,all of the groups intervened with an agent shared decreased levels of FBG,24 h U-mAlb,BUN,Scr and Cystatin C(P＜0.05,P＜0.01);alleviated renal pathological damage,and decreased levels of TNF-α,IL-1β and ICAM-1 in renal tissue and serum(P＜0.01).There existed no group difference between the Astragali Radix group and the C.sinica roots group in terms of all indices levels(P＞0.05).The C.sinica roots plus Astragali Radix group demonstrated its superiority over either C.sinica roots group or Astragali Radix group in terms of all the indices levels(P＜0.05,P＜0.01).CONCLUSION C.sinica roots,Astragali Radix or their combination use can alleviate the renal pathological damage and improve the renal function of DKD rats through inhibiting the M1 macrophages,reducing the secretion of inflammatory factors,whose mechanism may lie in the inhibition of JNK/SAPK signal pathway activation.A better effect can be anticipated by the combination use of C.sinica roots and Astragali Radix.

AIM To investigate the impact of the combination use of Caragana sinica Radix and Astragali Radix on a rat model of diabetic kidney disease(DKD).METHODS The SD rats were randomly divided into the normal group,the model group,the Engelgin group,the Caragana sinica Radix group,the Astragali Radix group and the Caragana sinica Radix-Astragali Radix compatibility group,with 10 rats in each group.Following the successful establishment of a DKD model by unilateral amputate renal combined with intraperitoneal injection of streptozotocin(STZ),the corresponding gastric gavage of drugs were administered for 8 weeks.The rats had their 24 h urinary microalbumin(24 h U-mALB)detected at 0,4 and 8 weeks;their levels of Scr,BUN,CysC,MDA and SOD activity detected by ELISA;their renal ROS expression detected by fluorescence probe method;their renal pathological changes observed by HE,PAS,Masson and PASM-Masson staining;their renal expressions of NOX4,Drp1,MFN2 and P62 detected by immunohistochemistry;and their renal expressions of PINK1,MFN2,Parkin,LC3-Ⅱ/Ⅰ,P62 and p-Drp1 proteins detected by Western blot.RESULTS Compared with the model group,each treatment group displayed lower contents of 24 h U-mALB,BUN,Scr and CysC in the serum of rats(P＜0.01);reduced pathological structure damage of the renal tissue;decreased MDA level in serum and kidney(P＜0.01);increased SOD activity(P＜0.01);increased renal protein expressions of PINK1,MFN2,Parkin and LC3-Ⅱ/Ⅰ(P＜0.05,P＜0.01);and decreased protein expressions of p-Drp1 and P62(P＜0.01).And the Astragali Radix group and the Caragana sinica Radix-Astragali Radix compatibility group took the lead(P＜0.05,P＜0.01).CONCLUSION Upon the rat model of DKD,the compatibility of Caragana sinica Radix and Astragali Radix may alleviate their renal pathological damage and improve their renal function by activating the mitochondrial autophagy to improve mitochondrial dynamics and inhibit their oxidative stress via PINK1/MFN2/Parkin pathway.

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

The study investigated the effect of Buyang Huanwu Decoction(BYHWD) on endogenous biomarkers in the urine of rats with chronic inflammation induced by lipopolysaccharide(LPS) using ultra-high performance liquid chromatography-quadrupole-time-of-flight-mass spectrometry(UPLC-Q-TOF-MS), aiming to elucidate the molecular mechanism underlying the therapeutic effect of BYHWD on chronic inflammation from a metabolomics perspective. Male SD rats were randomly divided into a normal group, a model group, and low-, medium-, and high-dose BYHWD groups(7.5, 15, and 30 g·kg~(-1)). The model group and BYHWD groups received tail intravenous injection of LPS(200 μg·kg~(-1)) on the first day of each week, followed by oral administration of BYHWD once a day for four consecutive weeks. Urine samples were collected at the end of the administration period, and UPLC-Q-TOF-MS was used to analyze the metabolic profiles of the rat urine in each group. Multivariate statistical analysis methods such as principal component analysis(PCA), partial least squares-discriminant analysis(PLS-DA), and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to analyze the effect of BYHWD on endogenous metabolites. One-way ANOVA and variable importance for the projection(VIP) were used to screen for potential biomarkers related to chronic inflammation. The identified biomarkers were subjected to pathway and enrichment analysis using MetaboAnalyst 5.0. A total of 25 potential biomarkers were screened and identified in the rat urine in this experiment. Compared with the normal group, the model group showed significant increases in the levels of 14 substances(P<0.05) and significant decreases in the levels of 11 substances(P<0.05). BYHWD was able to effectively reverse the trend of most endogenous biomarkers. Compared with the model group, BYHWD significantly down-regulated 13 biomarkers(P<0.05) and up-regulated 10 biomarkers(P<0.05). The metabolic products were mainly related to the biosynthesis of pantothenic acid and coenzyme A, tryptophan metabolism, retinol metabolism, and propionate metabolism. BYHWD has therapeutic effect on chronic inflammation induced by LPS, which may be related to its ability to improve the levels of endogenous metabolites, enhance the body's anti-inflammatory and antioxidant capabilities, and restore normal metabolic activity.
Rats ; Male ; Animals ; Chromatography, High Pressure Liquid/methods* ; Lipopolysaccharides ; Rats, Sprague-Dawley ; Metabolomics/methods* ; Inflammation/drug therapy* ; Biomarkers/urine*

OBJECTIVES: To explore the cytotoxicity of four wild mushrooms involved in a case of Yunnan sudden unexplained death (YNSUD), to provide the experimental basis for prevention and treatment of YNSUD. METHODS: Four kinds of wild mushrooms that were eaten by family members in this YNSUD incident were collected and identified by expert identification and gene sequencing. Raw extracts from four wild mushrooms were extracted by ultrasonic extraction to intervene HEK293 cells, and the mushrooms with obvious cytotoxicity were screened by Cell Counting Kit-8 (CCK-8). The selected wild mushrooms were prepared into three kinds of extracts, which were raw, boiled, and boiled followed by enzymolysis. HEK293 cells were intervened with these three extracts at different concentrations. The cytotoxicity was detected by CCK-8 combined with lactate dehydrogenase (LDH) Assay Kit, and the morphological changes of HEK293 cells were observed under an inverted phase contrast microscope. RESULTS: Species identification indicated that the four wild mushrooms were Butyriboletus roseoflavus, Boletus edulis, Russula virescens and Amanita manginiana. Cytotoxicity was found only in Amanita manginiana. The raw extracts showed cytotoxicity at the mass concentration of 0.1 mg/mL, while the boiled extracts and the boiled followed by enzymolysis extracts showed obvious cytotoxicity at the mass concentration of 0.4 mg/mL and 0.7 mg/mL, respectively. In addition to the obvious decrease in the number of HEK293 cells, the number of synapses increased and the refraction of HEK293 cells was poor after the intervention of Amanita manginiana extracts. CONCLUSIONS The extracts of Amanita manginiana involved in this YNSUD case has obvious cytotoxicity, and some of its toxicity can be reduced by boiled and enzymolysis, but cannot be completely detoxicated. Therefore, the consumption of Amanita manginiana is potentially dangerous, and it may be one of the causes of the YNSUD.
Humans ; HEK293 Cells ; Sincalide ; China ; Amanita ; Death, Sudden

Objective:To study the insulin-like growth factors-1 (IGF-1) and lipid level of term small for gestational age (SGA) infants within 24 hours postnatally and to explore the correlation between IGF-1 and blood lipids.Methods:A prospective study was conducted on singleton term SGA and appropriate for gestational age infant (AGA) who were delivered and admitted to the neonatal ward of Guangdong Women and Children Hospital within 24 hours after birth from May 2020 to January 2021, and the infants were divided into SGA and AGA groups to compare the differences in IGF-1 and lipid levels within 24 hours after birth and to analyze the correlation between IGF-1 and lipids.Results:A total of 95 cases in the SGA group and 84 cases in the AGA group were included in the study. The proportion of infants with IGF-1 <25 ng/ml was significantly higher in SGA group (87.4%) than in the AGA group (52.4%). It was also found that the proportion of infants with IGF-1 <25 ng/ml in SGA was significantly higher than that in AGA within different gender composition groups, early-term and full-term births groups. The triglyceride (TG) level was higher in the SGA group than that in the AGA group, but the high-density lipoprotein cholesterol (HDL-C) level was lower than that in the AGA group ( P<0.05). IGF-1 level within 24 hours postnatally in SGA and AGA was positively correlated with HDL-C levels ( P<0.01) and negatively correlated with TG ( P<0.01), and HDL-C level was a predictor of IGF-1. Conclusions:Compared with term AGA, SGA term infants showed insufficient IGF-1 and HDL-C secretion and high TG within 24 hours after birth. Nutritional support for SGA should be given promptly after birth to avoid hypoglycemia and to stimulate IGF-1 secretion.

Objective    To systematically review risk factors for esophagogastric anastomotic leakage (EGAL) after  esophageal cancer surgery for adults to provide theoretical basis for clinical prevention and treatment. Methods    PubMed, Web of Science, The Cochrane Library, WanFang Data, VIP, CNKI and CBM were searched from inception to January 2020 to collect case control studies and cohort studies about risk factors for EGAL after esophageal cancer surgery. Two reviewers independently screened literature, extracted data and assessed risk of bias of included studies, and then, meta-analysis was performed by using RevMan 5.3 software. Results    A total of 33 studies were included, including 19 case-control studies and 14 cohort studies, all of which had a Newcastle-Ottawa Scale (NOS)≥6. There were 26 636 patients, including 20 283 males and 6 353 females, and there were 9 587 patients in China and 17 049 patients abroad. The results of meta-analysis showed that the following factors could increase the risk for EGAL (P≤0.05), including patient factors (18): age, sex, body mass index (BMI), smoking history, smoking index (≥400), alcohol history, digestive tract ulcer, respiratory disease, lower ratio of forced expiratory volume in one second to forced vital capacity (FEV1/FVC), chronic obstructive pulmonary disease (COPD), coronary atherosclerosis, peripheral vascular disease, arrhythmia, diabetes, hypertension, cerebrovascular disease, celiac trunk calcification and descending aortic calcification; preoperative factors (6): abnormal liver function, renal insufficiency, American Society of Anesthesiologists (ASA) grading, neoadjuvant radiotherapy and preoperative albumin<35 g/L, preoperative lower albumin; intraoperative factors (7): retrosternal route, cervical anastomosis, thoracoscopic surgery, operation time≥4.5 h, tubular stomach, upper segment tumor, splenectomy; postoperative factors (5): respiratory failure, postoperative arrhythmia, use of fiberoptic bronchoscopy, pulmonary infection, deep venous thrombosis. Neoadjuvant chemotherapy could reduce the risk for postoperative EGAL (P<0.05). However, age≥60 years, upper gastrointestinal inflammation, diffusing capacity for carbon monoxide (DLCO%), thoracic surgery history, abdominal surgery history, glucocorticoid drugs history, neoadjuvant chemoradiotherapy, anastomotic embedding, end-to-end anastomosis, hand anastomosis, intraoperative blood loss and other factors were not significantly correlated with EGAL. Conclusion    Current evidence suggests that the risk factors for postoperative EGAL include age, sex, BMI, smoking index, alcohol history, peptic ulcer, FEV1/FVC, COPD, diabetes, ASA grading, neoadjuvant radiotherapy, preoperative albumin<35 g/L, cervical anastomosis, thoracoscopic surgery, operation time≥4.5 h, tubular stomach, upper segment tumor, intraoperative splenectomy, postoperative respiratory failure, postoperative arrhythmia and other risk factors. Neoadjuvant chemotherapy may be the protection factor for EGAL. Due to limited study quality, more high quality studies are needed to verify the conclusion.

Objective:To disclose the molecular genetic differences of Culex flavivirus among mosquitoes in Gansu province in 2011 and 2019.Methods:Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to obtain the nucleotide sequences of Culex flavivirus genomes from mosquitoes in Gansu province in 2011 and 2019, and viral molecular biology and bioinformatics method were used to analyze the genetic differences of the viruses.Results:Nucleotide sequences of 10 strains of Culex flavivirus were obtained, including 8 strains (all from Culex pipiens pallens) obtained in 2011 and 2 strains ( from Culex tritaeniorhynchus and Anopheles sinensis) in 2019. Homology analysis of nucleotide and amino acid sequences of virus E gene showed that the nucleotide sequence similarity and amino acid similarity of viruses isolated from Gansu in 2019 and 2011 ranged from 98.3%-100% and 95.4%-97.3%, respectively. Phylogenetic analysis of Culex flavivirus E gene sequence showed that two strains of Culex flavivirus isolated in Gansu province in 2019 (GS1975 and GS1976) and eight strains of Culex flavivirus isolated in 2011 all belonged to group B of genotype 1 of Culex flavivirus. Further analysis found that GS1975 virus isolated in 2019 was in a common evolutionary cluster with viruses isolated from Liaoning (2010 and 2011) and Inner Mongolia (2018), while GS1976 virus isolated in 2019 formed a coevolutionary cluster with viruses isolated from Inner Mongolia (2018) and Gansu (2011). Conclusions:Although both Culex flaviviruses isolated in Gansu province in 2011 and 2019 are genotype 1 virus, the two viruses isolated in 2019 distributed in two different evolutionary clusters, suggesting that the local mosquito virus genome changes over time, therefore, long-term monitoring of molecular differences is needed to carry out.