OBJECTIVETo observe the effect of natural type ginsenoside Rg2 (Rg2) and its stereoisomers [20 (R)-Rg2 and 20 (S)-Rg2] at different concentrations on oxygen-glucose deprivation/ reperfusion (OGD/R) induced cortical neuronal injury model in vitro, and to explore the mechanism, and compare their differences of action.

METHODSCortical neurons after 7-day culture were randomly divided into 5 groups, i.e., the control group, the model group, the Rg2 group, 20 (R) -Rg2 group, and 20 (S) - Rg2 group. Cortical neurons in the Rg2 group, 20 (R)-Rg2 group, and 20(S)-Rg2 group were pretreated with 20, 40, and 80 μmol/L Rg2, 20 (R) -Rg2, and 20 (S) -Rg2 for 24 h to prepare OGD/R model. The cell survival rate, the activity of Caspase-3, the intracellular Ca2+ concentration, contents of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected 24 h later.

RESULTSCompared with the control group, cell survival rates and activities of SOD obviously decreased, the activity of Caspase-3, Ca2+ fluorescent optical gray value, and contents of MDA significantly increased with statistical difference (P < 0.05). Compared with the model group, cell survival rates and activities of SOD obviously increased, the activity of Caspase-3, Ca2+ fluorescent optical gray value, and contents of MDA significantly decreased in 20 μmol/L Rg2 group, 40 μmol/L 20 (R) -Rg2 group, and 80 μmol/L 20 (S) -Rg2 group (P < 0.05). Compared with 20(S)-Rg2 group, cell survival rates increased and contents of MDA significantly decreased in 20, 40, and 80 μmol/L Rg2 and 20 (R)-Rg2 groups (P < 0.05). The activity of Caspase-3 decreased and contents of SOD increased in 80 μmol/L 20 (R)-Rg2 group, and 40, 80 μmol/L Rg2 groups (P < 0.05). Ca2+ fluorescent optical gray value decreased in 40, 80 μmol/L Rg2 and 20 (R)-Rg2 groups (P < 0.05). Compared with 20 (R)-Rg2 group, Ca2+ fluorescent optical gray value decreased in 80 μmol/L Rg2 group (P < 0.05); contents of SOD increased in 40 and 80 μmol/L Rg2 groups (P < 0.05); contents of MDA decreased in 20, 40, and 80 μmol/L Rg2 groups (P < 0.05).

CONCLUSIONSRg2 and its stereoisomers could improve cell vitality of cortical neurons against OGD/R induced injury. This might be related to improving anti-apoptotic capacities and antioxidant abilities, and reducing Ca2+ inflow. Besides, the neuroprotective effect of 20 (R) -Rg2 was better than that of 20 (S) -Rg2, but inferior to that of Rg2.

Antioxidants ; metabolism ; Apoptosis ; Calcium ; metabolism ; Caspase 3 ; metabolism ; Cell Survival ; Cells, Cultured ; Ginsenosides ; pharmacology ; Glucose ; Humans ; Malondialdehyde ; metabolism ; Neurons ; drug effects ; Neuroprotective Agents ; pharmacology ; Oxygen ; Random Allocation ; Reperfusion Injury ; Stereoisomerism ; Superoxide Dismutase ; metabolism

Objective To evaluate the effects of retrograde colonic electrical stimulation (RCES) with trains of short pulses and RCES with long pulses on colonic transit in irritable bowel syndrome (IBS) rats and to investigate whether stress-induced visceral hypersensitivity could be alleviated by RCES so as to find a valuable new approach for IBS treatment. Methods A total of 48 male rats were randomly divided into model group and control group. Visceral hypersensitivity model was induced by a 6-day HIS protocol composed of two stressors, restraint stress for 40 min and forced swimming stress for 20 min. The extent of visceral hypersensitivity was quantified by electromyography and abdominal withdrawal reflex scores (AWRs) of colorectal distension (use a balloon) at different pressures. After the modeling, all rats were equipped with electrodes in descending colon for retrograde electrical stimulation and a PE tube for perfusing phenol red saline solution in the ileocecus. After recovering from surgery, RCES with long pulses, RCES with trains of short pulses, and sham RCES were performed in colonic serosa of rats for 40 min in six groups of 8 each, including three groups of visceral hypersensitivity rats and three groups of health rats. Colonic transit was assessed by calculating the output of phenol red from the anus every 10 min for 90 min. Finally, the extent of visceral hypersensitivity will be quantified again in model group. Results After the 6-day HIS protocol, the HIS rats displayed an increased sensitivity to colorectal distention, compared to control group at different distention pressures (P < 0.01). CRES with trains of short pulses and long pulses significantly attenuated the hypersensitive responses to colorectal distention in the HIS rats compared with sham RCES group (P < 0.01). The effects of RCES on rats colon transmission: In the IBS rats, the colonic emptying were (77.4 ± 3.4)%, (74.8 ± 2.4)% and (64.2 ± 1.6)% in the sham RCES group, long pulses group and trains of short pulses group at 90 min; In healthy rats, The colonic emptying was (65.2 ± 3.5)%, (63.5 ± 4.0)% and (54.0 ± 2.5)% in the sham RCES group, long pulses group and trains of short pulses group at 90 min. Conclusion RCES with long pulses and RCES with trains of short pulses can significantly alleviate stress-induced visceral hypersensitivity. RCES with trains of short pulses has an inhibitory effect of colonic transit, both in visceral hypersensitivity rats and healthy rats.

OBJECTIVETo study the antiinflammatory effect of a compound TCM (Traditional Chinese Medicine) agent on animal models. The agent contains ant extractive and a blent of three herbal products, herba epimedii, fructus cnidii, and fructus lycii.

METHODThree animal models to induce experimental inflammation in rats, including carrageenin--induced paw edema, cotton-ball granuloma and adjuvant induced arthritis, were chosen to study the antiinflammatory effect of the TCM agent.

RESULTThe TCM agent showed a marked inhibitory effect on edema induced by all three types of inflammation in rats, the inhibitory rate of the TCM agent at the dose of 0.20, 0.40 and 0.80 g.kg-1 in granuloma model bing over 25% at 1 hour post oral administration, and being 23.8%, 22.7%, 39.7% at 6 hour. In addition, the TCM agent also showed a significant preventive as well as therapeutic effect on adjuvant induced arthritis in rats, and improved the pathological changes of the animal joints with the induced arthritis.

CONCLUSIONTCM agent has significant antiinflammatory effects on the three above mentioned animal models.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Ants ; Arthritis ; drug therapy ; Capsules ; Cnidium ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; therapeutic use ; Edema ; drug therapy ; Epimedium ; chemistry ; Granuloma, Foreign-Body ; drug therapy ; Lycium ; chemistry ; Male ; Materia Medica ; therapeutic use ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar

The aim of this study was to investigate the effect of cytochrome C on apoptosis of HL-60 cells and to explore the mechanism of HL-60 cell apoptosis induced by cytochrome C. The HL-60 cells were treated with different concentrations of cytochrome C for 24 hours. The concentrations of cytochrome C were as follows: 0 (control group), 9.375 mg/L, 18.75 mg/L, 37.5 mg/L, 75 mg/L and 150 mg/L. The apoptosis was analyzed by flow cytometry (FCM) after HL-60 cells were treated with different concentrations of cytochrome C for 24 hours. The fas mRNA expression changes of relevant apoptotic genes were examined by reverse transcription-polymerase chain reaction (RT-PCR) after HL-60 cells were treated with different concentrations of cytochrome C for 24 hours and were analyzed half-quantitatively. The expressions of fas involved in HL-60 treated with different concentrations of cytochrome C for 24 hours were detected by Western blot. The results indicated that the flow cytometry (FCM) (PI straining) showed obvious hypo-diploid peak before G(1) phase in the treated group, and its apoptotic ratio increased in a dose-dependent manner. The fas mRNA expression levels of the relevant apoptotic genes could be detected by RT-PCR after HL-60 cells treated with different concentrations of cytochrome C for 24 hours, and the expression of fas mRNA in HL-60 cells was increased by cytochrome C in dose-dependent manner within range of 0-37.5 mg/L. It is concluded that the cytochrome C up-regulates expressions of fas mRNA and their protein so as to induce apoptosis of the HL-60 cells.
Apoptosis ; drug effects ; Cytochromes c ; pharmacology ; Dose-Response Relationship, Drug ; HL-60 Cells ; Humans ; RNA, Messenger ; metabolism ; Up-Regulation ; fas Receptor ; metabolism

OBJECTIVETo evaluate the progression in morphologic changes of lungs in SARS patients.

METHODSFour cases of SARS with lung tissue samples available (including one for ultrastructural examination) were enrolled into the study. Histochemical study for VG, Masson, reticulin, orcein, PAS, sirius red stains and immunohistochemical study for vimentin, desmin, smooth muscle actin, HHF-35, CD34, F8, collagen types I and III were also performed.

RESULTSAccording to the morphologic changes, lung lesions in SARS were subcategorized into 3 phases: acute exudative inflammation, fibrous proliferation and the final fibrotic stage. Two cases belonged to the acute exudative phase, in which the course was less than 20 days. The principal lesions consisted of acute alveolar exudative inflammation, hyperplasia of alveolar epithelium, necrosis, alveolar hyaline membrane formation, alveolar desquamation and focal fibroplasia. The acute exudative protein was PAS-positive. There was an increase in reticulin fiber formation. The reactive fibroblasts were highlighted by desmin and vimentin. One case belonged to the fibroproliferative stage, in which the course was around 25 days. Major lesions included proliferative interstitial pneumonia with early pulmonary fibrosis. There was also evidence of organizing pneumonia, with an increase in reticulin fiber formation, which had a glomeruloid appearance on special stain. The mesenchymal cells showed either myofibroblastic (which expressed desmin, HHF-35, smooth muscle actin and vimentin) or fibroblastic (which expressed vimentin only) differentiation. Fibroelastosis and fibroplasia was also noted. The remaining case belonged to the fibrotic stage, in which the course was around 75 days. The main features included diffuse fibrosis and honeycomb change, which were highlighted by sirius red stain. Immunohistochemistry showed mainly types I and IV collagen fibers. In all lesions, there was also an increase of number of CD68-positive macrophages.

CONCLUSIONSThe morphologic progression in lungs of SARS patients is characterized by the development of increased fibrosis. The primitive mesenchymal cells, hyperplastic alveolar epithelial cells and macrophages play an important role in the pathogenesis.

Actins ; metabolism ; Adult ; Collagen Type I ; metabolism ; Desmin ; metabolism ; Humans ; Lung ; metabolism ; pathology ; Male ; Middle Aged ; Pulmonary Fibrosis ; pathology ; Severe Acute Respiratory Syndrome ; metabolism ; pathology ; Vimentin ; metabolism

BACKGROUNDThe N400 component of event-related potentials (ERP) has recently drawn widespread attention at home and abroad. This study was to explore the relationship between N400 changes and risperidone treatment and rehabilitation infirst-episode schizophrenia (FES).

METHODSERP component N400 was recorded by Guangzhou Runjie WJ-1 ERP instruments, in 58 FES before and 6 months, 15 months after risperidone treatment, and in 62 normal controls. The patients' syndromes were assessed by Positive and Negative Syndrome Scale (PANSS). And the stimuli are Chinese sentences with matching (congruent) or mismatching (incongruent) ending words.

RESULTSN400 latencies were prolonged, and amplitudes were decreased in Cz, Pz, Fz, C3, C4, in FES compared with in NC, before treatment. The prolonged N400 latencies and decreased amplitudes were negatively correlated with the patients' positive scale and total scale of PANSS. There are significant differences of N400 amplitudes and latencies in 6 months and 15 months follow-up after treatment. Before treatment, 6 months and 15 months after treatment, N400 latencies are 446 ± 35 ms, 440 ± 37 ms, 414 ± 31 ms (F = 9.72, P < 0.01) in incongruent situation; N400 amplitudes are 5.2 ± 4.6 μV, 5.7 ± 4.8 μV, 7.3 ± 5.0 μV (F = 2.06, P > 0.05) in congruent situation, and 8.5 ± 5.9 μV, 10.1 ± 5.0 μV, 11.9 ± 7.0 μV (F = 3.697, P < 0.05) in incongruent situation.

CONCLUSIONSN400 could be used to predict the effects of treatment of schizophrenia to some degree. The linguistic and cognitive impairment in schizophrenia can be improved by antipsychotic drugs.

Adult ; Evoked Potentials ; Follow-Up Studies ; Humans ; Middle Aged ; Risperidone ; therapeutic use ; Schizophrenia ; drug therapy ; rehabilitation

Abstract：Objective To predict the structure and function of sterol O⁃acyltransferase 1（SOAT1）related to hepatocellular carcinoma（HCC）by using bioinformatics tools，in order to understand its mechanism as the marker and therapeutic target of S⁃Ⅲ subtype. Methods The structure，function and protein interaction of SOAT1 were predicted and analyzed by using databases or softwares such as NCBI，STRING，Protscale，SignalP，TMHMM，PSORT，SOPMA，SWISS ⁃ MODEL， NetNGlyc，NetOGlyc，Netphos and ProtParam. Results The protein encoded by SOAT1 was a hydrophobic protein with good stability，which was a nonclassical pathway protein with 8 transmembrane regions，mainly distributed among the cell membrane. SOAT1 was expressed in many tissues，while most of them in the adrenal gland，which showed multiple phosphorylation sites and was mainly involved in the synthesis and catabolism of cholesterol. Conclusion Bioinformatics analysis of structure and function of SOAT1 showed that SOAT1 lipid synthesis and catabolism pathways played an important role，and lipid expression was closely related to the development of cancer，indicating that the treatment of HCC may be achieved by regulating the expression of SOAT1 gene.

Aim To analyze the molecular mechanism of rhubarb in the treatment of aeute pancreatitis ( AP) by network pharmacology and molecular docking.Methods TCMSP,TCMID and Swiss target predic¬tion databases were used to screen the active compo¬nents and targets of rhubarb,and genecards and OMIM databases were used to screen the targets of AP.Then the active ingredient drug target network of rhubarb and theactive ingredient disease target network of rhubarb for AP were constructed by using Cytoscape software.PPI network was constructed in string database, and go and KEGG enrichment analysis was performed in metascape database and R language.Finally,molecular docking was used to verify the possibility of binding the core active components to the core target.Results A total of 192 active components and 1 882 AP targets were obtained.The first three active components of rhubarb in the treatment of AP were beta sitosterol, aloe emodin and eupatin.The core target of rhubarb in the treatment of AP was hsp90aal.Go enrichment analysis focused on reaction to toxic substances, while KEGG enrichment analysis was significantly enriched in p53 signaling pathway closely related to AP.Molecular docking showed good binding and stable conformation.Conclusions Rhubarb can affect the expression of AP related genes and proteins through p53 signaling pathway, thereby inhibiting cell apoptosis and allevia¬ting the inflammatory injury of AP.

OBJECTIVETo clarify the roles of SPK pathway in the regulation of proliferation, survival and glucose consume of human erythroleukemia TF-1 cells.

METHODSThe interfering in SPK expression of TF-1 cells was performed using leutivirus vector-mediated shRNA, the interference efficacy of SPK in TF-1 cells was detected by RT-qPCR and Western blot, the viability of TF-1 cell proliferation was detected by using CCK-8 method, the apoptosis of TF-1 cells was determined by flow cytmetry with Annexin V staining.

RESULTSHypoxia up-regulated the expression of HIF-1α, HIF-2α, and SPK in TF-1 cells. SPK treatment resulted in reduced proliferation and induced apoptosis in TF-1 cells. Furthermore, knockdown of the SPK significantly reduced utilization and consumption of glucose.

CONCLUSIONThe SPK is key signalling molecule involved in regulation of hypoxia-induced proliferation and glucose metabolism in TF-1 cells, and plays an important rote in proliferation and energy metabolism of leukemia cells.

OBJECTIVETo investigate the effect and mechanism of miR-486 on glycometabolism of hematopoietic cells.

METHODSqRT-PCR was applied to detect the expression of miR-486 or Sirt1 on TF-1 cells under hypoxia. Lentivirus was used to mediate the overexpression or inhibition of miR-486 on TF-1 cells and qRT-PCR was used to detect the expressions of Sirt1, glucose transporter 1(Glut1) and glucose transporter 4(Glut4). Lentivirus-mediated Sirt1-shRNA transduction was used to knockdown Sirt1 expression which was detected by qRT-PCR and Western blot. The expressions of Glut1 and Glut4 were determined by qRT-PCR.

RESULTSHypoxia promoted the expression of miR-486 and inhibited the expression of Sirt1. MiR-486 overexpression could inhibit the expression of Sirt1 and promote the expressions of Glut1 and Glut4, whereas miR-486 silencing upregulated the sirt1 expression and inhibited the expressions of Glut1 and Glut4. And inhibition of Sirt1 expression increased the expressions of Glut1 and Glut4.

CONCLUSIONMiR-486 can regulate the glycometabolism of hematopoietic cells by targeting Sirt1.