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

Objective To estimate the validity of Charlson' s weighted index of comorbidities (WIC) used to predicting 28-day mortality among ICU pneumonia patients with underlying diseases.Methods Aretrospective analysis of 160 adult patients with pneumonia admitted to a multi-discipline ICU of Shanghai Changzheng hospital between October 2010 and February 2012 was carried out.Clinical data were collected including age,gender,community acquired pneumonia (CAP) or hospital acquired pneumonia (HAP),underlying diseases,severity-of-sepsis,and 28-day mortality.WIC scores,acute physiology and chronic health evaluation (APACHE) Ⅱ,and sepsis related organ failure assessment (SOFA) were assessed within the first 24 hours after admission.Logistic regression analyses were used to evaluate the predictors for outcome.The receiver operating characteristic curve (ROC) was used to compare the performance of these scores between different methods.Results Of 160 enrolled patients,76 (48.8％) were CAP,82 (51.2％) HAP,and 106 (66.3％) male,54 (33.7％) female,and 99 (61.9％) patients survived and 61 (38.1％) died.The average age was (62.4 ± 17.3) years old.Compared with survivors,WIC,APACHE Ⅱ and SOFA scores were significantly higher in death group (P ＜ 0.05).The multivariate logistic regression revealed that risk of death depends predominantly on age (OR =1.049,95％ CI:1.011-1.088,P =0.011),WIC (OR =1.725,95％ CI:1.194-2.492,P =0.004),APACHE Ⅱ score (OR =1.175,95％CI:1.058-1.305,P =0.003),SOFA score (OR =1.277,95％ CI:1.048-1.556,P =0.015),presence of ARDS (OR =0.081,95％ CI:0.008-0.829,P =0.034),and complicated with severe sepsis (OR =0.149,95％ CI:0.232-0.622,P =0.004).The area under the receiver operating characteristics curve in predicting mortality was 0.639 (0.547-0.730) for the WIC,0.782 (0.709-0.856) for APACHE Ⅱ score,0.79 (0.714-0.866) for SOFA score and 0.842 (0.777-0.907) for the merger of three.Conclusions In pneumonia patients of ICU,WIC is a useful approach to predicting 28-day mortality,and the risk of death significantly depends on co-morbidities.

Objective To observe the therapeutic effects of end-to-side and side-to-side portal vein-hepatic artery shunts (P-H shunt) in pigs with liver cirrhosis and portal hypertension.Methods There were 15 pigs in each group (control and experimental),and the P-H shunt was made in each animal to observe the process of blood flow into the liver and the reduction of portal vein blood pressure.Results The portal vein pressures before the P-H shunt of pigs in the control and experimental group were (20.51±0.74) cm H2O (1 cm H2O=0.098 kPa) and (30.82±2.53) cm H2O respectively (P＜0.05).Portal vein pressures 30 days after the P-H shunt were(19.75±0.84) cm H2O and (20.84± 1.36) cm H2 O respectively (P＞0.05).The portal vein pressure differences of pigs in the control and experimental group before P-H shunt were (7.20±0.34) cm H2O and (17.34±0.62) cm H2O respectively (P＜0.05).Pressures differences 30 days after the P-H shunt were (6.40±0.21) cm H2O and (7.84 ± 1.32) cm H2O respectively (P＜0.05).Splenic vein injection of methylene blue after the P-H shunt operation stained the liver well.No necrosis and hepatic encephalopathy occurred for 30 days after the P-H shunt,and the splenomegaly gradually returned to normal.Liver function was most affected after the operation and other biochemical markers were least affected.Conclusions The P-H shunt changed the channel of blood flow into the liver,the portal hemodynamic was not affected,and a new balanced mechanism was established to maintain the portal hemodynamic stability.However,further treatment was needed for liver function recovery.

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.

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 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.

OBJECTIVETo investigate the effects of Jiangu granule containing serum (JGG-serum) on the cyclins in rat's osteoblast at G1 phase.

METHODSOsteoblasts isolated by enzymatic digestion from SD rats were cultured and intervened with JGG-serum or normal saline (as control) respectively. Cell generation cycle was detected by flow cytometry, and expressions of Cyclin D1, cyclin-dependent kinase 4 (CDK4), oncogene protein (P21) in the osteoblast were detected dynamically using immuno-cytochemical and RT-PCR technique.

RESULTSAs compared with the control, the cell generation cycle and cell proliferation were proceeding quicker in the JGG-serum (20%) intervention group; with higher protein and mRNA expressions of Cyclin D1 and CDK4, as well as much lowered expressions of P21 in nuclei of osteoblast detected at all time points (24 h, 48 h and 72 h after treatment, P < 0.05 or P < 0.01).

CONCLUSIONJGG-serum can adjust the G1 phase cyclins in osteoblast cultured in vitro, increase the mRNA and protein expressions of Cyclin D1 and CDK4, and inhibit P21 expression, so as to accelerate the proliferation of osteoblast.

Animals ; Animals, Newborn ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; genetics ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; G1 Phase ; drug effects ; Male ; Osteoblasts ; cytology ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Serum

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