[Objective]To explore the degranulation effect and mechanism of Saposhnikovia divaricata(SD)extract on rat basophilic leukemia cell line RBL-2H3 cell.[Methods]Methylthialazole tetrazolium(MTT)test was used to select the concentrations in the subsequent experiments based on impact of 5,25,50,100,200,400 μg·mL-1 SD extract on the activity of RBL-2H3 cells.Immunoglobulin E(IgE)induction was used to establish RBL-2H3 cell degranulation model.Blank control group,model group,low dose SD extract group(5 μg·mL-1),medium dose SD extract group(25 μg·mL-1),high dose SD extract group(50 μg·mL-1)and dexamethasone(DXMS)group(100 μg·mL-1)were set up,with intervention for 30 minutes.MTT test was used to detect the effect of low,medium,high-dose SD extract on activity of RBL-2H3 cell degranulation model.Toluidine blue staining was used to observe the morphology of degranulation cells and calculate degranulation rates.Immunofluorescence staining was used to detect expression of F-actin.Enzyme linked immunosorbent assay(ELISA)was used to detect the levels of β-aminohexosidase,histamine,interleukin-4(IL-4),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ).Western blot was used to detect the expressions of phosphatidylinositide-3 kinase(PI3K),phosphorylation-PI3K(p-PI3K),protein kinase B(AKT),phosphorylation-AKT(p-AKT),p38 mitogen activated protein kinase(p38MAPK),phosphorylation-p38MAPK(p-p38MAPK),nuclear factors-κB(NF-κB),phosphorylation-NF-κB(p-NF-κB),extracellular regulated kinases(ERK)and phosphorylation-ERK(p-ERK)protein.[Results]The low,medium,high doses of SD extract(5,25,50 μg·mL-1)had no significant effects on the activity of RBL-2H3 cells(P＞0.05).Compared with blank control group,the number of toluidine blue stained cells of model group was decreased,cells shape rounded,degranulation rate was increased,expression of F-actin was decreased,the levels of β-aminohexosidase,histamine,IL-4,IL-6,TNF-α were increased,IFN-γ level was decreased,the expressions of p-PI3K/PI3K,p-AKT/AKT,p-p38MAPK/p38MAPK,p-NF-κB/NF-κB and p-ERK/ERK were increased(P＜0.01).Compared with model group,the F-actin expression of low,medium,high doses of SD extract groups and DXMS group was increased,levels of β-aminohexosidase,histamine,IL-4,IL-6 and TNF-α were decreased,IFN-γ level was increased(P＜0.01),the expressions of p-PI3K/PI3K,p-AKT/AKT,p-p38MAPK/p38MAPK,p-NF-κB/NF-κB and p-ERK/ERK were decreased(P＜0.05,P＜0.01);the number of toluidine blue stained cells in medium,high dose SD extract groups and DXMS group was increased with spindle cell shape,degranulation rate was decreased(P＜0.01).Compared with low dose SD extract group,degranulation rate of high dose SD extract group and DXMS group was decreased(P＜0.01),F-actin expression was increased(P＜0.05),p-p38MAPK/p38MAPK expressions were decreased(P＜0.01).[Conclusion]SD extract inhibited degranulation of IgE sensitized RBL-2H3 cell and decreased the levels of inflammatory mediators,its mechanism may be related to the inhibition the phosphorylation expression of PI3K/AKT,p38MAPK/NF-KB and ERK.

Abstract OBJECTIVE To investigate the effect and mechanism of flavonoids from Sophora flavescens Ait. on testicular tissue damage in male mice induced by local heat stress in the scrotum. METHODS TCMSP database was used to screen the targets of flavonoids in Sophora flavescens Ait., and the bioinformatics analysis was performed on the target. The mouse model of scrotal heat stress was used and the flavonoids of Sophora flavescens Ait. was used for intervention. The sperm density and sperm aberration rate of mice in each group were measured, and the morphological changes of testicular tissue were observed. Tumor necrosis factor-α (TNF-α) and interleukin 17(IL-17) mRNA and protein levels in testicalar were detected of by q-PCR and Western blotting. Na+-K+-ATPase, Mg2+-ATPase, Ca2+-ATPase, lactate dehydrogenase(LDH), sorbitol dehydrogenase(SDH) activity, malondialdehyde (MDA), NO, TNF-α level and the content of testosterone in serum were detected in tissue homogenate. RESULTS Heat stress could lead to the decrease of sperm density and increase of aberrant sperm, the obvious thinning of testicular spermatogenic epithelium, the decrease of cell level and quantity, the significant decrease of ATPase, LDH, SDH activities, and the increase of MDA, NO content, TNF-α and IL-17 expression in testicular tissue. After the intervention with 250, 500 mg·kg-1·d-1 flavonoids of Sophora flavescens Ait., the quality of sperm and the damage of testicular tissue morphology were improved. The level of TNF-α and IL-17 in serum and testicular tissue were decreased, and the activity of ATPase, SDH and the level of testosterone were increased. CONCLUSION The mechanism of flavonoids of Sophora flavescens Ait. is through inhibiting the inflammatory factor TNF-α and IL-17 levels, improve the anti-lipid peroxidation ability and inhibite the role of NO, enhance the activity of energy enzymes in spermatogenesis, improve the level of serum testosterone, and improve the reproductive disorders caused by heat stress.

Objective:To construct a nomogram model predicting the occurrence of acute kidney injury (AKI) in patients with sepsis in the intensive care unit (ICU), and to verify its validity for early prediction.Methods:Sepsis patients admitted to the ICU of the Affiliated Hospital of Jining Medical University from April 2015 to December 2021 were retrospectively included, and those who met the inclusion criteria were randomly divided into training and validation sets at a ratio of 7:3. Univariate and multivariate logistic regression models were used to identify independent risk factors for AKI in patients with sepsis, and a nomogram was constructed based on the independent risk factors. Calibration curve, receiver operating characteristic (ROC) curve, and decision curve analysis (DCA) were used to evaluate the nomogram model.Results:741 patients with sepsis were included in the study, 335 patients developed AKI within 7 d of ICU admission, with an AKI incidence of 45.1%. Randomization was performed in the training set ( n=519) and internal validation set ( n=222). Multivariate logistic analysis revealed that acute physiology and chronic health status score Ⅱ, sequential organ failure score, serum lactate, calcitoninogen, norepinephrine dose, urea nitrogen, and neutrophil percentage were independent factors influencing the occurrence of AKI, and a nomogram model was constructed by combining these variables. In the training set, the AUC of the nomogram model ROC was 0.875 (95% CI: 0.767-0.835), the calibration curve showed consistency between the predicted and actual probabilities, and the DCA showed a good net clinical benefit. In the internal validation set, the nomogram model had a similar predictive value for AKI (AUC=0.871, 95% CI: 0.734-0.854). Conclusions:A nomogram model constructed based on the critical care score at admission combined with inflammatory markers can be used for the early prediction of AKI in sepsis patients in the ICU. The model is helpful for clinicians early identify AKI in sepsis patients.

Objective:To investigate the effect of calcium carbonate combined with vitamin K on bone metabolism indexes, Th1 cytokines and safety in children with glucocorticoid-induced osteoporosis.Methods:A total of 96 children with glucocorticoid-induced osteoporosis were enrolled in this study. They were randomly divided into control group and study group, 48 cases in each group. The control group was treated with calcium carbonate, and the study group was treated with calcium carbonate combined with vitamin K. The bone metabolism index serum total bone type Ⅰ collagen N-terminal peptide was compared between the two groups before and after treatment. PINP), serum β-Carboxy I terminal peptide ( β-CTX), osteocalcin (osteocalcin, P<0.05), and serum β-CTX. BGP), serum bone alkaline phosphatase (BALP), Th1 cytokine interferon- γ (IFN- γ), interleukin-2 (IL-2) levels and adverse reactions were also observed. Results:There was no statistical significance in the general data of the two groups (all P>0.05). Compared with that before treatment, bone mineral density of femoral neck increased in both groups after treatment, and the improvement in the study group (-1.02±0.49) was more significant than that in the control group (-1.52±0.65) ( t=4.26, P<0.001). Compared with those before treatment, the levels of PINP, β-CTX, BGP and BALP in 2 groups after treatment were decreased, and compared with those of the those of the control group (PINP: 31.65±6.58; β-CTX: 0.34±0.05; BGP: 4.95±1.28; BALP: 40.54±7.84), all indexes of the study group after treatment (PINP: 26.54±7.06; β-CTX: 0.24±0.03; BGP: 3.05±1.09; BALP: 35.96±7.02) improved significantly ( t=3.67, P<0.001; t=11.88, P<0.001; t=7.83, P<0.001; t=3.02, P<0.003). Compared with that before treatment, IL-2 level was increased while IFN- γ level was decreased in both treatment groups. Additionally, in comparison to the control group (IL-2: 163.89±30.85; IFN- γ: 196.61±21.05), IL-2 level (198.32±32.14) was higher and the IFN- γ level (163.25±18.43) was lower in the study group after treatment ( t=5.35, P<0.001; t=8.26, P<0.001). The incidence of adverse reactions between the two groups was not statistically significant ( χ2=0.15, P=0.695) . Conclusion:Calcium carbonate combined with vitamin K in the treatment of children with glucocorticoid-induced osteoporosis can improve bone metabolism indexes and Th1 cytokine levels in children, and the clinical therapeutic effect is good.

OBJECTIVE: To analyze the risk factors of in-hospital death in patients with sepsis in the intensive care unit (ICU) based on machine learning, and to construct a predictive model, and to explore the predictive value of the predictive model. METHODS: The clinical data of patients with sepsis who were hospitalized in the ICU of the Affiliated Hospital of Jining Medical University from April 2015 to April 2021 were retrospectively analyzed,including demographic information, vital signs, complications, laboratory examination indicators, diagnosis, treatment, etc. Patients were divided into death group and survival group according to whether in-hospital death occurred. The cases in the dataset (70%) were randomly selected as the training set for building the model, and the remaining 30% of the cases were used as the validation set. Based on seven machine learning models including logistic regression (LR), K-nearest neighbor (KNN), support vector machine (SVM), decision tree (DT), random forest (RF), extreme gradient boosting (XGBoost) and artificial neural network (ANN), a prediction model for in-hospital mortality of sepsis patients was constructed. The receiver operator characteristic curve (ROC curve), calibration curve and decision curve analysis (DCA) were used to evaluate the predictive performance of the seven models from the aspects of identification, calibration and clinical application, respectively. In addition, the predictive model based on machine learning was compared with the sequential organ failure assessment (SOFA) and acute physiology and chronic health evaluation II (APACHE II) models. RESULTS: A total of 741 patients with sepsis were included, of which 390 were discharged after improvement, 351 died in hospital, and the in-hospital mortality was 47.4%. There were significant differences in gender, age, APACHE II score, SOFA score, Glasgow coma score (GCS), heart rate, oxygen index (PaO₂/FiO₂), mechanical ventilation ratio, mechanical ventilation time, proportion of norepinephrine (NE) used, maximum NE, lactic acid (Lac), activated partial thromboplastin time (APTT), albumin (ALB), serum creatinine (SCr), blood urea nitrogen (BUN), blood uric acid (BUA), pH value, base excess (BE), and K⁺ between the death group and the survival group. ROC curve analysis showed that the area under the curve (AUC) of RF, XGBoost, LR, ANN, DT, SVM, KNN models, SOFA score, and APACHE II score for predicting in-hospital mortality of sepsis patients were 0.871, 0.846, 0.751, 0.747, 0.677, 0.657, 0.555, 0.749 and 0.760, respectively. Among all the models, the RF model had the highest precision (0.750), accuracy (0.785), recall (0.773), and F1 score (0.761), and best discrimination. The calibration curve showed that the RF model performed best among the seven machine learning models. DCA curve showed that the RF model exhibited greater net benefit as well as threshold probability compared to other models, indicating that the RF model was the best model with good clinical utility. CONCLUSIONS The machine learning model can be used as a reliable tool for predicting in-hospital mortality in sepsis patients. RF models has the best predictive performance, which is helpful for clinicians to identify high-risk patients and implement early intervention to reduce mortality.
Humans ; Hospital Mortality ; Retrospective Studies ; ROC Curve ; Prognosis ; Sepsis/diagnosis* ; Intensive Care Units

OBJECTIVE： To establish the method for simultaneous determination of 69 kinds of pesticide residues in Paeonia tactilora， Astragalus membranaceus， Ranunculus ternatus and Cornus officinalis. METHODS： GC-MS/MS method was adopted. The determination was performed on HP-5MS fused silica capillary column with splitless injecting samples. The injector temperature was set at 240 ℃， and sample size was 1 μL. The carrier gas was high-purity helium， the inlet mode was constant pressure， the pre-column pressure was 146 kPa， and the temperature was programmed. Triple four-pole tandem mass spectrometry was used for the detection， and electron impact ion source was used as ion source. The temperature of the ion source was 230 ℃， ionization energy was 70 eV， and the collision gas was nitrogen. The inlet temperature was 280 ℃ and four-pole temperature was 150 ℃， mass spectrometry monitoring mode was multi-reaction monitoring （MRM）， and solvent delay time was 5 min. RESULTS： The linear range of 69 kinds of pesticide residue was 4.82-399.6 ng/mL （all r＞0.990）. LODs were all in the range of 0.001 7-0.013 3 mg/kg， and LOQs were all in the range of 0.000 5-0.004 mg/kg. RSDs of precision and stability tests were less than 10％ （n＝6）. RSD of reproducibility test was lower than 5％ （n＝6， only pesticide amidine and permethrin were detected）. The recoveries were in the range of 62.9％-123.5％ （all RSD＜10％， n＝6）. Among 12 batches of samples， dichlorvos and diphenylamine were detected in C. officinalis； chlordimeform and permethrin were detected in  A. membranaceus； diphenylamine and chlordimeform were detected in P. tactilora； diphenylamine and vinclozolin were detected in R. ternatus. CONCLUSIONS： The method is simple in operation and reproducible for simultaneous determination of 69 kinds of pesticide residue in   P. tactilora， A. membranaceus， R. ternatus and C. officinalis.

Objective To investigate the resistance mechanism and virulence genes of clinical strains of carbapenem-resistant Klebsiella pneumonia ( CRKP ) .Methods Twenty clinical CRKP strains were collected from Tianjin Medical University General Hospital during May 2014 and May 2015.Vitek-2 Compact system was used for identification of the strains and antibiotic susceptibility test .Modified Hodge Test and EDTA double disk phenotypic test were performed for screening of drug -resistant phenotypes .Drug-resistant genes , capsular serotypes and associated virulence genes were amplified by PCR , and positive products were sequenced and analyzed by DNA sequencing .Results Resistance to beta-lactam antibiotics including cephalosporins and carbapenems was observed in 80.0%and above strains , but more than 70.0%strains were sensitive to tigecycline , amikacin and levofloxacin .KPC gene and NDM gene were found in 7 strains (35.0%) and 8 strains (40.0%), respectively.SHV, the most common extended-spectrumβ-lactamases ( ESBLs ) gene, was found in 16 strains ( 80.0%). DHA plasmid-mediated AmpCβ-lactamase was found in 2 strains (10.0%).Deletions of porin-coding genes OmpK35 and OmpK36 were found in 8 stains ( 40.0%) and 13 strains ( 65.0%), respectively.Carbapenem-resistant genes in combination with ESBLs genes and/or variation of porin was found in 14 strains (70.0%), and ESBLs genes in combination with variation of porin was found in 4 strains (30.0%).Three strains were of capsular serotype K1 and 1 was of K57, and all of them carried KPC genes .Virulence gene rmpA was found in 8 strains and all carried carbapenemases , among which 5 strains with KPC, 2 strains with NDM, 1 strain with both KPC and NDM .Six strains were aerobactin gene positive , among which 4 strains carried KPC genes . FimH-1 was positive in all strains .Conclusions KPC and NDM genes mainly account for resistance in CPKP, and ESBLs with OmpK gene deletion may also be an important cause .Strains with capsular serotypes K1 and K57 carrying KPC genes are common .

Objective:To study the improvement effect of Sophora Flavones combined with captopril on the rat diabetic cardiomyopathy,and to clarify its mechanisms of improving the myocardial fibrosis.Methods:Fifteen rats were selected randomly from 100 male Wistar rats as normal group.The other rats were fed with high fat and high sugar food and intraperitoneally injected with small dose of streptozotocin (30 mg· kg-1 )all at once to establish type 2 diabetes mellitus cardiomgopathy models.Then 72 rat models with type 2 diabetic cardiomyopathy were set up in which 60 rats were selected according to the blood glucose levels and divided into model group, captopril group,Sophora Flavones group,and Sophora Flavones combined with captopril group (combination group) (n=15).8 weeks after intragastric administration,the weights,the cardiac indexes,the activities of serum creatine kinase-MB (CK-MB), lactate dehydrogenase (LDH ), and the contents of nitric oxide (NO), nitric oxide synthase (iNOS),collagen Ⅰ (Col Ⅰ)and collagen Ⅲ (Col Ⅲ)of the rats in various groups were detected. Results:Compared with normal group,the weight of the rats in model group was decreased and the cardiac index was increased (P <0.01),the activities of serum LDH and CK-MB were increased (P < 0.01),the contents of iNOS and NO in the myocardium tissue of the rats in model group were decreased (P <0.01),and the contents of Col Ⅰ and col Ⅲ were increased (P <0.05 or P <0.01).Compared with model group,the weights and the cardiac indexes of the rats in medication groups were decreased (P <0.05),the activities of serum LDH and CK-MB were decreased (P <0.05 or P <0.01),the contents of iNOS and NO in the myocardium tissue were increased (P <0.05),and the contents of Col Ⅰ and Col Ⅲ were decreased (P <0.05 or P <0.01).The effect of combination group was better than single medication groups (P < 0.05).Conclusion:Combination of Sophora Flavones and captopril has a improvement effect on the rat type 2 diabetic cardiomyopathy,and its mechanism may be related to reducing the myocardial injury to improve diabetic cardiomyopathy.

Aim To observe the expression of epider-mal growth factor receptor ( EGFR) in cerebral tissues around hematomas after intracerebral hemorrhage, and explore the effects of EGFR on activation of astrocytes derived from rats and the involved mechanisms. Meth-ods The specimens of cerebral tissues around hemo-tomas after intracerebral hemorrhage undergoing hemo-tomas removal operation were collected and then divid-ed into 4 groups according to the time of intracerebral hemorrhage: <1 d, 1 ~5 d, 6 ~10 d and >10 d groups. Each group included 20 cases. At the same time, 20 dropped brain tissues distant to hemorrhage in the operative process were collected as control group. Immunohistostaining and Western blot were used to measure the expression of EGFR. After isolation and culturing, the astrocytes of rat cortex were treated with culture solution ( control group) , CNTF that was used to activate astrocytes, scramble siRNA + CNTF and EGFR siRNA +CNTF for 24h, respectively. The ex-pression of glial fibrillary acidic protein ( GFAP) mR-NA was detected through fluorescence real-time quanti-tative PCR. In addition, the protein levels of GFAP, signal transducers and activators of transcription 3 ( STAT3 ) and phosphorylated STAT3 ( p-STAT3 ) were examined using Western blot. Results With the ex-tension of intracerebral hemorrhage time, positive sig-nal index and protein expression levels of EGFR gradu-ally elevated, reached the peak on 6 ~10d, and then decreased after 10 d. There was statistical difference ( P<0. 01 ) . The expression levels of GFAP mRNA and protein as well as p-STAT3 were significantly in-creased in cells treated with CNTF alone as compared to control group ( P <0. 01 ) , whereas these effects were almost completely reversed by EGFR siRNA transfection ( P <0. 01 ) . Additionally, there was no statistical difference in STAT3 protein levels among groups ( P >0. 05 ) . Conclusions EGFR expression is upregulated in the cerebral tissues around hemotomas after intracerebral hemorrhage. Gene silence of EGFR contributes to suppressing the activation of astrocytes derived from rats, which may be involved in the block-ade of STAT3 phosphorylation.

Objective To investigate the inhibitory effect of Rhodiola total flavonoids on transforming growth factor (TGF)-β1-induced proliferation of cardiac fibroblasts (CFB) in the rats, and to explore its mechanisms of improving myocardial fibrosis.Methods 5 μg·L-1 TGF-β1 was used to induce the proliferation of CFB to build the cell models of myocardial fibrosis.The cultured CFB were divided into control group,model group,and low, medium,high doses (25,50 and 100 mg· L-1 )of Rhodiola total flavonoids groups.The cells were treated for 48 h,the vitalities of cells were detected by MTT,the levels of collagen proteinⅠ (ColⅠ)and collagen proteinⅢ(Col Ⅲ)in supernatant were measured by ELISA,and the activities of total superoxide dismutase (T-SOD)and glutathione peroxidase (GSH-Px)were detected,the levels of malondialdehyde (MDA)and glutathione (GSH) were also measured.Results The MTT results indicated that compared with control group,the cell vitality in model group was significantly increased (P < 0.01 ).compared with model group,the cell vitalities of CFB in Rhodiola total flavonoids groups were significantly decreased (P < 0.05 ). Compared with control group, the T-SOD activity in model group was decreased, while the MDA level was significantly increased (P <0.05);compared with model group,the T-SOD activities in 50 and 100 mg· L-1 Rhodiola total flavonoids groups were increased (P <0.01);the MDA levels in 25 and 50 mg· L-1 groups were decreased significantly (P < 0.05). Compared with control group,the GSH-Px activity and GSH level in model group were significantly decreased (P <0.01).Compared with model group,the GSH-Px activities and GSH levels in Rhodiola total flavonoids groups were increased (P <0.05).Compared with control group,the ColⅠand Col Ⅲ levels in model group were significantly increased (P <0.01);compared with model group,the ColⅠand Col Ⅲ levels in Rhodiola total flavonoids groups were significantly decreased (P <0.05).Conclusion Rhodiola total flavonoids can inhibit the proliferation of rat CFB.The development of myocardial fibrosis may be inhibited by Rhodiola total flavonoids through anti-oxidative stress pathway.