Yinchenzhufu decoction (YCZFD) is a classic formula for treating Yin Huang syndrome, which can improve liver injury caused by cholestasis. However, the mechanism of action of YCZFD still remains unclear. This article used network pharmacology, molecular docking, animal experiments, and molecular biology methods to explore the mechanism of YCZFD in treating liver injury caused by cholestasis. A mouse model of acute cholestasis induced by lithocholic acid was used to investigate the effects of YCZFD on liver injury. The experimental procedures described in this paper were reviewed and approved by the Ethical Committee at the Shanghai University of Traditional Chinese Medicine (approval NO. PZSHUTCM190823002). The results showed that YCZFD could reduce the levels of blood biochemical indicators and improve hepatocyte damage of cholestatic mice. Then, multiple databases were used to predict the corresponding targets of YCZFD active components on cholestatic liver injury. An intersection target protein-protein interaction (PPI) networks based on String database and Cytoscape software was used to demonstrate the possible core targets of YCZFD against cholestatic liver injury. The results indicated that core targets of YCZFD include tumor necrosis factor, interleukin-1β, non-receptor tyrosine kinase Src, interleukin-6, etc. GO (gene ontology) and KEGG (kyoto encyclopedia of genes and genomes) enrichment analysis indicated that YCZFD may regulate the tumor necrosis factor signaling pathway, nuclear factor-κB signaling pathway, bile secretion, and other related factors to ameliorate the cholestatic liver injury. AutoDockTools software was used to perform molecular docking verification on the core targets and components of YCZFD. To verify the results of network pharmacology, UPLC-MS/MS method was used to determine the effect of YCZFD on levels of bile acid profiles in mouse liver tissues. It was found that treatment with YCZFD significantly reduced the content of free bile acids, taurine bound bile acids, and total bile acids in the liver tissues of cholestatic mice. Then, results from real time PCR and Western blot also found that YCZFD can upregulate the expression of hepatic nuclear receptor farnesoid X receptor, metabolizing enzyme (UDP glucuronidase transferase 1a1), and efflux transporters (bile salt export pump, multidrug resistance-associated protein 2, multidrug resistance-associated protein 3, etc) in cholestasis mice, promote bile acid metabolism and excretion, and improve bile acid homeostasis. Moreover, YCZFD can also inhibit pyroptosis and inflammation by regulating NOD-like receptors 3 pathway, thereby inhibiting cholestatic liver injury.

Objective To investigate the effect of bone mesenchymal stem cells on Toll-like receptors (TLR) 2/4 expression in the lungs of rats with acute hemorrhagic necrotizing pancreatitis (AHNP). Methods Seventy SD male rats were randomly divided into sham-operation group (n=10), AHNP group(n=30) and MSCs-treated group(n=30). Masc rate of BMSCs with surface mark were measured by flow cytometer. TLR2/4mRNA expression in the the lung were measured by RT-PCR, and The ratio of Wet/dry and lung histological changs were observed. Results TLR2/4 mRNA could be detected in the lungs with low values in sham-operation group, markedly increased in 3 h, and peaked in 12 h in AHNP group (P＜0.05). Lung injuries were aggravated and the levels of TNF-α in the lung were increased (P＜0. 05) . Treatment with MSCs could effectively inhibit TLR2/4 mRNA expression and relieve lung injuries. The levels of TNF-α in the lung were decreased (P＜0.05). Conclusions The expression of TLR2/4 mRNA is increased in the lungs in AHNP and the lung injuries are aggravated. MSCs could markedly inhibit TLR2/4 mRNA expression in the lungs in AHNP, which would lead to relief of lung injury.

Objective In this experiment,we explored the effects of rAAV\|hGDNF on protecting spinal neurons From death. Methods rAAV\|hGDNF particals were produced by recombinant virus technolgy,and infected the culture spinal neurons which were exposed to glutamate.We counted the mortality rate and detected the expression of NOS mRNA by RT\|PCR. Results In the group transfering rAAV\|hGDNF the death rate was inhibited(50%?0\^02,control 59\^25%?0\^023, P

To establish vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) as secretary biomarkers for cell growth on topographic substrates, we have evaluated the secretion and expression of these 2 factors by SH-SY5Y human neuroblastoma cells on poly-L-lactide (PLLA) micropillar arrayed topographic substrates. We fabricated topographic substrates with UV lithography, silicon etching and polydimethylsiloxane-based replica molding, and interfaced SH-SY5Y human neuroblastoma cells with both the topographic substrates and PLLA flat substrates. Cell morphology and spreading were examined with scanning electron microscopy. The secretion and mRNA expression of VEGF and IL-8 were evaluated with enzyme linked immunosorbent assay (ELISA) and real time qPCR, respectively, 24 hours after cell plating. We successfully achieved 4 topographic substrates with a nominal pillar diameter of 2 microm and 4 microm, and a nominal pillar spacing of 2 microm and 7 microm. We found that the secretion and mRNA expression of VEGF and/or IL-8 by SH-SY5Y cells on 2-2 microm (pillar diameter-spacing), 4-2 microm and 4-7 microm topographic substrates were upregulated in comparison to those by cells on PLLA flat substrate, 24 hours after cell plating. Furthermore, both cytokines were even more substantially upregulated on the 2-7 microm substrate than on the other 3 topographic substrates. Compared to those on PLLA flat substrate, cells on topographic substrates showed significant changes in morphology (spreading area, perimeter and roundness), and the increase in the secretion and mRNA expression of VEGF and IL-8 was accompanied with a decrease in cell spreading areas. These results provided evidence that pillar arrayed topography was an important microenvironmental factor in affecting VEGF and IL-8 expression or secretion, and VEGF and IL-8 might serve as important secretary biomarkers for growth on topographic substrates by SH-SY5Y cells.
Biomarkers ; Cell Line ; Cell Proliferation ; Cellular Microenvironment ; Humans ; Interleukin-8 ; genetics ; secretion ; Neuroblastoma ; secretion ; Polyesters ; chemistry ; RNA, Messenger ; genetics ; Vascular Endothelial Growth Factor A ; genetics ; secretion

OBJECTIVETo explore the effect of negative emotions on serum levels of adrenocorticotropic hormone (ACTH) and neuropeptide Y (NYP) in hepatitis B liver cirrhosis (HBLC) patients.

METHODSTotally 617 HBLC patients were assigned to the negative emotion group (415 cases) and the non-negative emotion group (202 cases) judged by negative emotions. Case numbers of various grading Child-Pugh were recorded in the two groups. Their liver functions were compared between the two groups. Serum levels of ACTH and NPY were detected using double antibody sandwich enzyme-linked immunosorbent assay (ELISA) in the two groups.

RESULTSThere was no statistical difference in Child-Pugh grading between the two groups (χ2 = 0.65, P = 0.72). Compared with the non-negative emotional group, serum ACTH levels decreased significantly in the negative emotion group with statistical difference (P < 0.05). There was no statistical difference in serum ACTH levels between the two groups (P > 0.05).

CONCLUSIONThe negative emotion of HBLC patients was not related to the serum ACTH level, but to relatively lower-concentration serum NPY levels.

Adrenocorticotropic Hormone ; blood ; Emotions ; Hepatitis B ; blood ; psychology ; Humans ; Liver Cirrhosis ; blood ; psychology ; Neuropeptide Y ; Serum

Objective To analyze the differential expression of Stathmin in human cells infected with human-tropic porcine endogenous retrovirus(PERV)and to explore the potential molecular effect of human-tropic PERV on human cells.Methods HEK293 cells were infected with the human-tropic PERV infectious molecular clone.PCR,real-time RT-PCR and immunofluorescence analysis were applied to confirm that HEK293 cells were infected.Then real-time RT-PCR and Western blot were carried out to analyze the differential expression of Stathmin at the mRNA level and protein level,respectively.Results HEK293 cells were infected by human-tropic PERV.Real-time RT-PCR and Western blot analysis showed that Stathmin was up-regulated in HEK293 cells infected with PERV compared with the control cells.Conclusion Stathmin was up-regulated in HEK293 cells infected with human-tropic PERV.These studies will be helpful for revealing the interaction of PERV and human cells,and for understanding the molecular effect of humantropic PERV on human cells.In addition,it suggested that PERV infection may infect cell growth and physiological functions,even be pathogenic.These will help to clarify the biologic characteristics of PERV and evaluate the safety of PERV in pig to human xenotransplantation.

Objective To study the prognostic predictive value of quantitative dectroencephalography (qEEG)for patients with large middle cerebral artery infarction (LMCAI).Methods The scores of routine electroencephalography (EEG),qEEG and the Glasgow Coma Scale (GCS) of the patients within 72 hours after symptom onset were recorded.The short-term prognosis (death or survival) was evaluated at 1 month after the onset.The long-term prognosis (good or poor) was evaluated at 3 months after the onset.All the observed data in each prognostic group were compared.Results A total of 105 patients were included in the study.There were significant differences in the margin of amplitude integrated electroencephalogram (aEEG) (upper margin:19.11 ± 7.80 μV vs.11.87 ±6.41 μV;t =2.392,P =0.019; lower margin:11.90 ± 4.78 μV vs.7.58 ± 4.15 μV; t =3.327,P =0.022),Synek-classification (x2 =48.114,P =0.000) between the short-term survival group and the death group; in patients with left LMCAI,there were significant differences in the absolute energy of the β-activity (13.16 ±12.66 μV2 vs.19.20 ±17.96 μV2;t =-2.781,P =0.039),spectral edge frequency 95％ (SEF95％) (9.17 ± 3.24 Hz vs.10.36 ± 3.76 Hz; t =-5.614,P =0.002) between the short-term survival group and the death group.There were significant differences in the age (59.33 ±13.67 years vs.68.87± 10.473 years; t =-3.215,P =0.002),GCS scores (10.86±2.80 vs.9.21 ±2.51;t =2.511,P =0.015),SEF95％ (13.80 ±5.40 Hz vs.10.93 ±4.68 Hz; t ＝2.311,P =0.024) and sides of infarction (x2 =4.737,P =0.030) between the long-term good prognosis group and the poor prognosis group.Conclusion qEEG can be used as an effective means of monitoring for evaluating the prognosis of patients with LMCAI.

ObjectiveTo assess the diagnosis and treatment of staphylococcal scalded skin syndrome (SSSS)in children.MethodsA total of 177 children whh SSSS were eligible for this study.Clinical data were retrospectively reviewed to assess the patients' age and geographical areas,predisposing factors,prodrome,clinical features,results of laboratory examination(including bacterial isolation and drug susceptibility test),treatment and prognosis.ResultsThe median age at onset was 23 days past 15 months.Infants aged 1 to 3 years were the most commonly affected population.The male to female ratio was 8:9.SSSS occurred in summer or autumn in 71.2％ of the patients.Most patients came from countryside.Of these patients,148 (83.6％) had no definite inducement,13 (7.3％) had fever or cold history before the occurrence of SSSS.Skin lesions often began in the face(71.6％).Almost all cases presented with erythema and blisters.Abnormal myocardial enzymes were observed in 104(80.6％) of 129 patients,and Staphylococcus aureus was isolated from 38.7％ of 142 patients.Susceptibility testing of 21 Staphylococcus aureus isolates from patients in the past 3 years showed that 95.2％ and 95.0％ were sensitive to cephalosporins and amoxicillin/clavulanic acid respectively.Double-antibiotic therapy was effective in 72.9％ of the patients.Intravenous gamma globulin was given to 36.2％ of the patients in the early stage.Myocardial-protecting drugs were also administrated early in patients with abnormal myocardial enzymes.All the patients healed without scarring.ConclusionsFor infants with acute facial erythema and exfoliative skin lesions,SSSS should be suspected and timely antibiotic treatment should be given.Cephalosporins and semi-synthetic penicillins are effective and the first choice for the treatment of SSSS.Meanwhile,early addition of gamma globulin and myocardial-protecting drugs may be beneficial to disease recovery.

Objective To investigate the change of serum Nogo-A protein in patients with acute closed brain injury, and explore its relationship with the severity of neuronal damage and prognosis. Methods Thirty-one patients with acute closed brain injury were enrolled. Venous blood samples (2 mL) were obtained 1, 3 and 5 d after injury. Serum concentrations of Nogo-A protein were determined by ELISA. Patients were divided into mild (n =7), moderate (n = 10) and severe (n = 14) injury groups according to Glasgow coma score (GCS), and were divided into favorable prognosis (n = 23) and poor prognosis (n = 8) groups according to Glasgow outcome score (GOS). Another 20 healthy adults were served as controls. Results The mass concentrations of serum Nogo-A protein in mild, moderate and severe injury groups 1, 3, 5 d after injury were significantly higher than those in control group (P < 0.01), and the mass concentrations of serum Nogo-A protein in moderate and severe injury groups 1, 3, 5 d after injury were significantly higher than those in mild injury group (P <0.05, P <0.01). The mass concentrations of serum Nogo-A protein 1, 3, 5 d after injury were significantly higher in poor prognosis group than those in favourable prognosis group (P < 0.01). Conclusion Serum Nogo-A protein level significantly increases after brain injury, and is related to the degree of injury and prognosis.

Objective To observe the synthesis of TLR2 protein and mRNA expression in Kupffer cells(KCs). Methods BALB/C mice were used to make model of partial hepatic ischemia/reperfusion (I/R) injury. After injury, KCs were isolated with two-steps in situ perfusion technique. The synthesis of TLR2 protein was determined by flow cytometric analysis (FCM) and real time reverse transcription(polymerase) chain reaction (Real Time RT-PCR) was used to analyse its gene expression in KC. Results The expression of TLR2mRNA and synthesis of protein in KC were significantly higher in experimental group, compared with sham group (protein expression: 9.19?0.43 % vs 1.52 ?0.24%, P