Objective To observe the effect of Astragalus injection on the expressions of inflammatory cytokines in human primary macrophages stimulated by lipoteichoic acid (LTA) and lipopolysaccharide (LPS),and investigate its effects on inflammatory reactions of Gram-positive (G+) and Gram-negative (G-) bacteria sepsis and its mechanisms.Methods Percoll density gradient centrifugation was used to isolate the human peripheral blood mononuclear cells,then they were purified by immune Anti-Biotin Microbeads with magnetic character and under the induction of recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF),the cells were cultivated for 12 days in vitro,eventually the human monocyte-derived macrophage was formed.The cultured human macrophages were inoculated in 96-well plates (each group 3 wells) and 6-well plates (each group 3 wells).The cells were divided into control group (200 μL DMEM added in each well),LTA 1 mg/L group,LPS 0.1 mg/L group and low astragalus injection (0.1 mg/L) and high astragalus injection (0.2 mg/L) dose groups.After the incubator plates were put in an incubator for 24 hours,the protein content of IL-8 and IL-10 in supernatant were detected by enzymelinked immunosorbent assay (ELISA),and the mRNA expression levels of IL-8 and IL-10 were detected by real time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR).Results LTA and LPS all can obviously up-regulate the expression levels of pro-inflammatory factor IL-8 and anti-inflammatory factor IL-10 of macrophage.The expressions of IL-8 and IL-10 protein and mRNA in LTA group and LPS group were significantly higher than those in control group after cuhure for 8 hours and 24 hours,the degrees of increment were more significantly at 24 hours [LTA stimute group:IL-8 protein (ng/L,× 103):41.57± 1.90 vs.1.58 ±0.24,IL-8 mRNA (A value):21.49±8.05 vs.1.00±0.16;IL-10 protein (ng/L):5.90±3.02 vs.2.91 ± 1.54,IL-10 mRNA (A value):1.35±0.34 vs.0.95±0.14;LPS stimute group:IL-8 protein (ng/L,× 103):345.00±22.80 vs.5.60±0.31,IL-8 mRNA (A value):29.84 ± 8.93 vs.1.00 ± 0.16,IL-10 protein (ng/L):122.37 ± 39.26 vs.44.79 ± 3.67,IL-10 mRNA (A value):7.38 ± 1.58 vs.1.35 ± 0.34,all P ＜ 0.05].The Astragalus injection could regulate LTA and LPS to stimulate the macrophage to decrease the expression levels of pro-inflammatory factor IL-8 protein and mRNA and increase the expression levels of anti-inflammatory factor IL-10 protein and mRNA in the macrophage;the changes of regulatory effect in the 24 hour-culture of Astragalus injection high dose group was the most significant [LTA stimute group:IL-8 protein (ng/L,×103):22.63±1.91 vs.41.57±1.90,IL-8 mRNA (A value):12.10±1.93 vs.21.49±8.05,IL-10 protein (ng/L):14.03±2.22 vs.5.90±3.02,IL-10 mRNA (A value):10.37±6.08 vs.1.35±0.34;LPS stimute group:IL-8 protein (ng/L,× 103):167.75 ± 19.90 vs.345.01 ±22.80,IL-8 mRNA (A value):15.61 ± 3.63 vs.29.84±8.93;IL-10 protein (ng/L):243.22±14.41 vs.122.37±39.26,IL-10 mRNA (A value):16.14±4.10 vs.7.38± 1.58,all P ＜ 0.05].Conclusions In the process of inflammatory response,the pro-inflammatory and anti-inflammatory factors co-exist simultaneously.Astragalus injection can inhibit the expression levels of pro-inflammatory factor gene and protein in the inflammatory response of G+ and G-bacteria sepsis and in the mean time,it can promote the expression levels of anti-inflammatory factor gene and protein,thus the immune mechanism of sepsis is affected,achieving the balance between pro-inflammation and anti-inflammation.

Objective:To analyze the risk factors and clinical characteristics of liver injury in patients with sepsis and to provide a reference for early recognition, early diagnosis, early intervention, and improve the survival rate of patients.Methods:The clinical data of sepsis patients admitted to the department of general intensive care unit (ICU) of the Second Affiliated Hospital of Zhejiang University School of Medicine from July 2014 to October 2020 were retrospectively analyzed. According to the occurrence of acute liver injury, patients with sepsis were divided into the liver injury group and the non-liver injury group, and the differences of demographic data, history, history of primary diseases, laboratory indicators on the first time of admission, treatments, the severity of the disease and other indicators were compared and analyzed. Logistic regression was used to analyze the risk factors for sepsis-related liver injury.Results:A total of 527 patients with sepsis were enrolled, and 129 patients with acute liver injury, accounting for 24.48%. Compared with the non-liver injury group, acute physiology and chronic health evaluation Ⅱ (APACHEⅡ), sequential organ failure assessment (SOFA), pro-brain natriuretic peptide (pro-BNP), serum MB isoenzyme of creatine kinase (CK-MB), total bile acid (TBA), serum creatinine (SCr), blood urea nitrogen (BUN), lactic acid (Lac), lactate dehydrogenase (LDH), C-reactive protein (CRP), procalcitonin (PCT) in liver injury group were significantly increased [APACHEⅡ score: 23.00±10.40 vs. 16.10±8.10, SOFA score: 9.17±4.29 vs. 5.90±3.12, pro-BNP (ng/L): 5 500.0 (1 166.0, 16 865.0) vs. 1 377.2 (448.8, 6 136.5), CK-MB (U/L): 23.0 (13.0, 55.0) vs. 18.0 (13.0, 31.0), TBA (μmol/L): 5.0 (2.4, 12.9) vs. 2.6 (1.4, 4.9), SCr (μmol/L): 146.0 (75.0, 222.0) vs. 71.0 (52.0, 125.8), BUN (mmol/L): 13.4 (8.8, 20.2) vs. 7.9 (4.9, 11.6), Lac (mmol/L): 2.0 (1.4, 4.4) vs. 1.4 (1.0, 2.2), LDH (μmol·s -1·L -1): 6.43 (3.76, 11.99) vs. 4.55 (3.38, 6.63), CRP (mg/L): 113.0 (61.8, 201.0) vs. 95.0 (37.3, 170.1), PCT (μg/L): 3.8 (1.0, 23.3) vs. 0.8 (0.2, 6.4)], prothrombin time (PT), international standard ratio (INR) and activated partial thrombin time (APTT) were significantly longer [PT (s): 19.4±7.6 vs. 16.0±4.0, INR: 1.7±1.0 vs. 1.3±0.5, APTT (s): 54.0±25.8 vs. 44.1±15.1], plasma fibrinogen (FIB), platelet count (PLT), albumin (ALB), and cholesterol (CHOL) were decreased [FIB (g/L): 4.2±2.3 vs. 4.9±1.8, PLT (×10 9/L): 116.3±74.3 vs. 182.7±108.6, ALB (g/L): 25.4±5.5 vs. 27.6±5.5, CHOL (mmol/L): 2.5±1.2 vs. 3.2±1.3], the probability of shock was significantly increased (91.47% vs. 59.19%), and the duration of shock was prolonged [days: 5.0 (2.0, 9.0) vs. 1.0 (0.0, 3.0)], positive rate of microbial culture (81.40% vs. 71.11%), probability of occurrence of drug-resistant bacteria (67.44% vs. 47.99%) were significantly higher, mechanical ventilation time [days: 6.0 (2.0, 12.7) vs. 2.4 (0.0, 6.9)], continuous renal replacement therapy (CRRT) time [days: 1.2 (0.0, 5.0) vs. 0.0 (0.0, 0.0)], the length of intensive care unit (ICU) stay [days: 9.0 (5.0, 18.0) vs. 7.0 (3.0, 13.0)] were significantly longer, 28-day mortality was significantly higher (80.62% vs. 28.89%), and the differences were statistically significant (all P < 0.05). Further Logistic regression analysis showed that PLT decline, PT prolongation, CRRT duration, shock duration and 28-day mortality were correlated with sepsis-related liver injury [odds ratios ( OR) and 95% confidence interval (95% CI) were 0.992 (0.987-0.998), 3.103 (1.507-6.387), 1.198 (1.074-1.336), 1.196 (1.049-1.362), and 0.213 (0.072-0.633), respectively, all P < 0.05]. Conclusions:Prolonged PT and decreased PLT are independent risk factors for sepsis complicated with liver injury. The long duration of CRRT, long duration of shock, and high mortality are independent clinical characteristics of patients with sepsis-related liver injury.

[ ABSTRACT] AIM:To observe the effect of ginsenoside Rb 1 on the proliferation and the expression of serotonin transporter (SERT), 5-hydroxytryptamine 1B receptor (5-HT1BR) in rat pulmonary artery smooth muscle cells (PASMCs) under hypoxia condition and the relationship with Rho /Rho-kinase signal pathway .METHODS: PASMCs were isolated from the adult male SD rats and primarily cultured .The subcultured cells from the 4th generation to the 6th generation were harvested and divided into normal group , and hypoxia group , different concentrations of Rb 1 incubation groups treated with 50, 100 and 200 mg/L ginsenoside Rb1 under hypoxia (HR50, HR100 and HR200 groups, respectively).The viability of the PASMCs was measured by CCK-8 assay.BrdU positive cells were determined using flow cytometry .The expression of serotonin transporter and 5-HT1BR at mRNA and protein levels was detected by RT-PCR and Western blot, respectively. The PASMCs were randomly divided into normal group , hypoxia group , HR200 group and hypoxia +Y-27632 incubation group ( HY group ) .The mRNA expression of Rho-kinase and phosphorylated myosin phosphatase target subunit 1 ( p-MYPT1) protein level were investigated by RT-PCR and Western blot, respectively.RESULTS: Compared with normal group, the proliferation of PASMCs in hypoxia group was significantly increased (P<0.01).The cell viability and the ex-pression of SERT and 5-HT1B R at mRNA and protein levels in all different concentrations of Rb 1 groups were obviously de-creased compared with hypoxia group ( P<0.05 ) .The mRNA expression of Rho-kinase and protein level of p-MYPT1 were markedly decreased in HR200 group, and no significant difference compared with HY group was observed ( P <0.01).CONCLUSION: Treatment with ginsenoside Rb1 might prevent hypoxia-induced proliferation of PASMCs and over-expression of SERT and 5-HT1B R through inhibiting the Rho/Rho-kinase pathway .

Objective:To establish an early prediction model with multiple indicators to predict the risk of severe acute pancreatitis (SAP) in hyperlipidemic acute pancreatitis (HLAP).Methods:The clinical data of 92 patients with HLAP admitted to the Emergency Department of our hospital from March 2018 to February 2020 were analyzed retrospectively. Among them, 29 cases deteriorated to SAP and 63 cases did not. Univariate analysis was used to screen predictive indicators related to hyperlipidemic severe acute pancreatitis (HL-SAP), and logistic regression analysis was used to screen independent predictive indicators related to HL-SAP. Then a prediction model was established. The area under (AUC) the receiver operating curve (ROC) was used to evaluate the predictive ability of each predictive indicator and the model for HL-SAP. Bootstrap resampling technology was used to validate the predictive ability of the model.Results:Univariate analysis showed that procalcitonin, D-dimer, C-reactive protein, albumin, cholesterol and CT grade had influence on the progression of HLAP to SAP ( P<0.05). Logistic regression analysis showed that D-dimer ( OR=2.112, 95% CI: 1.022-4.366; P<0.05), CT grade ( OR=5.818, 95% CI: 2.481-13.643; P<0.01) and cholesterol ( OR=1.146, 95% CI: 1.004-1.308; P<0.05) were independent risk factor of HL-SAP. The AUC of D-dimer, CT grade, cholesterol and the model were 0.802, 0.875, 0.665 and 0.927, respectively. Internal validation of the predictive ability of the model showed that the C-index was 0.927. Conclusions:In the early phase, application of the prediction model that composes D-dimer, CT grade and cholesterol has a good predictive effect on HL-SAP.

To assess relationships between xanthine oxidase (XOD) and nephropathogenic infectious bronchitis virus (NIBV) infection, 240 growing layers (35 days old) were randomly divided into two groups (infected and control) of 120 chickens each. Each chicken in the control and infected group was intranasally inoculated with 0.2 mL sterile physiological saline and virus, respectively, after which serum antioxidant parameters and renal XOD mRNA expression in growing layers were evaluated at 8, 15 and 22 days post-inoculation (dpi). The results showed that serum glutathione peroxidase and superoxide dismutase activities in the infected group were significantly lower than in the control group at 8 and 15 dpi (p < 0.01), while serum malondialdehyde concentrations were significantly higher (p < 0.01). The serum uric acid was significantly higher than that of the control group at 15 dpi (p < 0.01). In addition, the kidney mRNA transcript level and serum activity of XOD in the infected group was significantly higher than that of the control group at 8, 15 and 22 dpi (p < 0.05). The results indicated that NIBV infection could cause the increases of renal XOD gene transcription and serum XOD activity, leading to hyperuricemia and reduction of antioxidants in the body.
Antioxidants ; Chickens ; Glutathione Peroxidase ; Hyperuricemia ; Infectious bronchitis virus* ; Kidney ; Malondialdehyde ; RNA, Messenger* ; Superoxide Dismutase ; Uric Acid ; Xanthine Oxidase* ; Xanthine*

Objective:To express virus-like particles of poliovirus type 2 (PV2-VLP) in insect cells using a recombinant baculovirus expressing P1 and 3CD and to preliminarily evaluate its immunogenicity.Methods:Based on the codon preference of High 5 cells, the sequences of P1 gene and 3CD gene of PV2 were optimized and inserted into pUC57-Amp to construct pUC57-PV2-P1 and pUC57-PV2-3CD. UC57-PV2-P1s mutant that carried P1 gene mutation affecting thermostability was then constructed. Recombinant baculovirus strains of rBac-PV2-P1s-3CD and rBac-PV2-P1-3CD (wild type) were constructed using homologous recombination. The expression of target proteins was detected by Western blot. PV2-VLP was purified by ion exchange chromatography. The structure of VLP was observed under transmission electron microscopy to evaluate the assembly efficiency. The immunogenicity of PV2-VLP was assessed in a rat model.Results:The recombinant baculovirus with stable expression of P1s and 3CD proteins was successfully constructed. Western blot results showed that the yield of VLP was higher after thermostability mutation than that of the wild type. A three-dimensional structure with a diameter of about 30 nm was observed under electron microscopy, indicating that the VLP was successfully assembled. Animal experiment showed that the recombinant PV2-VLP had immunogenicity and could effectively induce the production of neutralizing antibodies.Conclusions:Effective VLP vaccines could be successfully prepared using the insect cell-baculovirus expression system, which provided reference for the development of polio VLP vaccine.

OBJECTIVE: To determine the prevalence and incidence of illness of two-week duration, and the factors influencing these, among residents 15 years and older in four counties of Hunan Province. METHODS: Data were sampled from four counties of Hunan Province for the Fourth National Health Service Survey. Incidence and two-week prevalence of disease were used to assess the health service needs of residents. A non-conditional, stepwise logistic regression was employed to explore the influencing factors. RESULTS: The two-week prevalence and incidence were 11.5% and 3.9%, respectively, in four counties of Hunan. The three leading diseases of two-week prevalence were: respiratory diseases, digestive diseases, and musculoskeletal diseases. Non-conditional stepwise logistic regression showed that urban residents had 0.64 times the risk of two-week illness compared with the rural residents (P< 0.05); residents in the 45-59 year age group and the 60+ year age group had 1.69 and 2.62 times the risk of two-week illness compared with residents in the 15-44 year age group, respectively (P<0.05). The widowed had 1.91 times the risk of prevalence of two-week illness contrasted to singles (P<0.05); the students had 0.29 times the risk of two-week illness contrasted to the workers (P<0.05); urban residents had 0.63 times the risk of two-week illness compared with the rural (P<0.05); the widowed had 2.37 times the risk of incidence of two-week illness compared with singles (P<0.05). CONCLUSION The majority of health service needs of residents of four counties is generated by three diseases: respiratory diseases, digestive diseases, and musculoskeletal diseases. Relatively, rural residents, the elderly, employed persons and the widowed have higher health service needs than others and deserve specific attention.
Adolescent ; Adult ; Aged ; China ; epidemiology ; Community Health Services ; statistics & numerical data ; Digestive System Diseases ; epidemiology ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Musculoskeletal Diseases ; epidemiology ; Respiratory Tract Diseases ; epidemiology ; Sampling Studies ; Surveys and Questionnaires ; Young Adult

Objective:To explore the characteristics of T lymphocyte subsets and cytokines in hyperlipidemia-induced acute pancreatitis (HLAP) and its prognostic value.Methods:This study included 184 patients with acute pancreatitis (AP) admitted to the First Affiliated Hospital of Xiamen University from January 2018 to May 2021. Based on disease etiology, there were 92 HLAP cases and 92 non-hyperlipidemia-induced AP (NHLAP) cases. Stratified by disease severity according to 2012 Atlanta classification criteria, the patients were divided into the severe subgroup (SAP) and non-severe subgroup (NSAP). Peripheral venous blood samples were taken from all patients on day 1, 3, and 5 after admission. T lymphocyte subsets were determined by flow cytometry, and cytokines were detected by flow fluorometry. The number of CD4 +% and CD8 +% and the expression of cytokines were compared by Student’s t test or Mann-Whitney U analysis. Logistic regression analyses were performed to identify risk factors for severe AP, and a receiver operating characteristic (ROC) curve was constructed to predict severe AP. Statistical significance was taken as P<0.05. Results:Compared with the NHLAP group, patients in the HLAP group had lower CD4 +%, while higher levels of IL-2 on day 1 ( P<0.05), and had also lower CD4 +%, while higher levels of IL-4, IL-6, and IL-10 on day 3 ( P<0.05). Furthermore, IL-6 and IL-10 levels of the HLAP group were significantly increased compared to the NHLAP group on day 5 ( P<0.05). IL-10 levels in the SAP subgroup were significantly higher than those in the NSAP subgroup on day 1 ( P<0.05). Compared with the NSAP subgroup, the SAP subgroup had elevated levels of IL-2, IL-4, IL-6, IL-10 and IFN-γ on day 3 (all P<0.05), and had lower CD4 +%, while increased levels of IL-6 and IL-10 on day 5 (all P<0.05). Multivariate Logistic regression analysis showed that IL-10 was an immune indicator of independent risk factor for severe AP in the HLAP group on day 1 ( OR=1.139, 95% CI: 1.038-1.251, P<0.05). Finally, ROC analysis showed that the area under the curve of IL-10 to assess HLAP with severe AP was 0.772, and the best cut-off value for predicting severe AP was 5.6 pg/mL, with a sensitivity of 83.3% and a specificity of 68.8%. Conclusions:Changes of CD4 +% and cytokines are different between the HLAP and NHLAP groups. IL-10 can be used as a predictor of early disease severity in patients with HLAP.

Narciclasine(NCS),a hymenocallis littoralis alkaloid extracted from the bulbs of the genus Narcissus in the Lycoriaceae family,has been proven to have significant anti-tumor activity against a variety of tumor cells.The antitumor mechanisms of NCS are diverse and NCS exhibits antitumor effects through different pathways,which adapts to the current trend of developing multi-target anti-tumor drugs.This review introduces the research progress of the anti-tumor activity and mechanism of NCS in recent years based on the inhibitory effect of NCS on gastric cancer cells,oral cancer cells,polymorphous glioblastoma cells,colon cancer cells,breast cancer cells,melanoma cells and primary exudative lymphoma cells,aiming to provide ideas and references for the research and development,and design of NCS type anti-tumor drugs in the future.

Clinical data of a 33-year-old rheumatoid arthritis patient accompanied with pregnancy and lactation-associated osteoporosis(PLO) was retrospectively reviewed.The patient had two pregnancies within three years,and bone pain occurred during lactation of the second pregnancy after 2 months of delivery.Dual-energy X-ray (DEXA) showed decreased bone density,while thoracolumbar spine and pelvic X-ray film suggested osteoporosis.Her rheumatoid arthritis was stable and previous treatment did not include glucocorticoids.Other causes of metabolic bone diseases were excluded and the diagnosis of PLO was confirmed.The breastfeeding was stopped;calcium,active vitamin D,alendronate sodium were administrated,and the treatment was effective.Domestic and international literature from January 2000 to January 2019 was searched and no cases of PLO accompanied with rheumatoid arthritis were reported.There were only 100 cases of PLO retrieved in the literature,including 19 cases of PLO in China reported by four articles.It is suggested that PLO should be considered in RA patients presenting with arthragia during the lactation,in addition to relapse of disease.Awareness of PLO should be increased in physicians,especially rheumatologists,and anti-osteoporosis therapy is effective in patients with PLO.