This study is to investigate the role of endogenous CSE/H2S in regulating apoptosis of HepG2 cells. MTT and Trypan blue assay were performed to determine the effect of CSE inhibitor PAG and CSE siRNA on proliferation of HepG2. Production of H2S from HepG2 cells was assessed spectrophotometrically using N, N-dimethyl-p-phenylenediamine-dihydrochloride. Cells apoptosis was detected by means of double staining of Hoechst 33342 and PI with Array Scan V(TI)HCS600 High-Contents. Dihydroethidine (DHE) and 2', 7'-dichlorodihydrofluorescein diacetate (DCFH-DA) assay was used to determine intracellular superoxide anion and ROS level. Reduced glutathione (GSH) was determined by OxiSelect Total Glutathione Assay Kit. Recombinant plasmid pcDNA 3.1/myc-His(-)-CSE was constructed and transfected into 293T cells to rescue the ROS and GSH level to further investigate the effect of CSE/H2S on ROS and GSH. Western blotting was performed to test the effect of CSE siRNA on expression of activated caspase 3 and p-AKT and Nrf2 protein. The results showed that PAG and CSE siRNA could significantly decrease the production of H2S in HepG2 cells and inhibit the proliferation of HepG2 cells at a dose-dependent and time-dependent manner, respectively. PAG and CSE siRNA could promote the cell apoptosis of HepG2 cells. Moreover, PAG and CSE siRNA induced increased ROS generation and depletion of the critical antioxidant GSH and recombinant plasmid pcDNA 3.1/myc-His(-)-CSE rescued the level of ROS and GSH. Meanwhile, CSE siRNA increased the expression of activated caspase 3, but CSE siRNA did not affect the expression of p-AKT and Nrf2. These results suggested that the CSE/H2S pathway was involved in suppression of HepG2 cell growth and promoted apoptosis of HepG2 cells in an oxidative stress-dependent manner.

Objective To compare the effectiveness,safety and related clinical indicators between simple drainage treatment and drainage treatment combined with intrathoracic urokinase for children with parapneumonic pleural effusion(PPE).Methods Twenty-nine in patients with PPE given pleural effusion drainage in the First Affiliated Hospital of Zhengzhou University from January 2013 to December 2015 were selected as research subjects,who were divided into a simple group and an urokinase group based on whether intrathoracic urokinase was injected or not.The total number of hospital stay,the total drainage volume,the total number of catheter days,the total cost,the days with fever,efficient rate,operation rate and security of the patients were retrospectively analyzed between two groups.Results The intrathoracic days of hospital stay [M(P25,P75)] of urokinase group[19(11,30) days]were less than those of simple group[30(21,38) days],and the difference was significant (Z =-2.545,P =0.011);the total drainage volume[M(P25,P75)] of the urokinase group [430 (175,1 308) mL] was more than that of the simple group [110 (10,325)mL],and the difference was significant (Z =-2.811,P =0.005);the total number of catheter days [M (P25,P75)] of urokinase group [9 (7,19) days] was less than that of the simple group [20 (10,30) days],and the difference was significant (Z =-2.020,P =0.043);the total cost [M(P25,P75)] of the urokinase group [20 000(10 000,30 000)RMB] was less than that of the simple group [40 000 (30 000,50 000) RMB],and the difference was significant (Z =-2.631,P =0.009);the days with fever between urokinase group and the simple group was not significant (Z =-0.820,P =0.412).The urokinase group had a higher cure rate[76.9％ (10/13 cases)] and a lower surgical rate [23.1％ (3/13 cases)] compared with those of the simple group[18.7％ (3/16 cases),81.3％ (3/16 cases)],and the difference was significant (x2 =9.814,P =0.003).Conclusions Intrapleural urokinase therapy as an adjuvant treatment of PPE is simple and convenient,economic,higher efficiency,lower risk,which can be used as an effective clinical solution such disease.

AIM: To investigate the expression and effects of sialyl Lewis X (SLeX) on the invasion and migration of human hepatocellular carcinoma HepG2 cells.METHODS: The expression of α1,3-fucosyltransferase VII (FUT7) in HepG2 cells and L-02 cells was detected by RT-qPCR and Western blot.The SLeX expression in HepG2 cells and L-02 cells was determined by Western blot and immunocytochemical staining.The invasion and migration abilities of the treated cells were evaluated by Transwell assay.RESULTS: The expression of FUT7 and SLeX in the HepG2 cells, but not in the L-02 cells, was observed.The invasion rates of the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L were significantly decreased as compared with control group (P<0.05).The migration ability of the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L was also significantly reduced as compared with control group (P<0.05).The invasion rate and migratory cell number were significantly different between any 2 groups in the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L (P<0.05).CONCLUSION: HepG2 cells express SLeX.SLeX is closely related to the migration and invasion abilities of the HepG2 cells.

This study is to investigate the inhibitory effect and mechanism of prosapogenin A (PSA) on MCF7. MTT assay was performed to determine the inhibitory effect of PSA on MCF7 cells. PI/Hoechst 33342 double staining was used to detect cell apoptosis. RT-PCR was used to test the mRNA levels of STAT3, GLUT1, HK and PFKL. Western blotting was performed to determine the expression of STAT3 and pSTAT3 protein in MCF7 cells. The results showed that PSA could dose-dependently inhibit cell growth of MCF7 followed by IC50 of 9.65 micrmol x L(-1) and promote cell apoptosis of MCF7. Reduced mRNA levels of STAT3, HK and PFKL were observed in MCF7 cells treated with 5 micromol x L(-1) of PSA. PSA also decreased the level of pSTAT3 protein. STAT3 siRNA caused decrease of mRNA of GLUT1, HK and PFKL which indicated STAT3 could regulate the expressions of GLUT1, HK and PFKL. The results suggested that PSA could inhibit cell growth and promote cell apoptosis of MCF7 via inhibition of STAT3 and glycometabolism-related gene.

Objective To explore the potential of autologous dendritic cells (DC) pulsed with HLA-A201-binding peptide E613-21(KLPDLCTEL) and E786-94(TLGIVCPI)in inducing specific T cells respouse in vitro.Methods Cervical carcinoma patients with positive HLA-A201 were enrolled and their monocytes isolated and induced into dendritic cells and pulsed with HLA-A201-binding peptide E613-21 and E786-94.PBLs were primed by DCs every week for thee times.The cytokine level of supernatant of CTLs was tested by ELISA.The percentage of special CTLs was tested by flow cytometry.The specific killing effect of CTLs was tested by MTT.Results the numbers of DCs of eleven cervical carcinoma patients were (10.79±0.88) ×106(100 ml peripheral blood).CDllc+HLA-DR+(97.15±2.41)％,CD80+(84.28+5.39)％,CD83 +(85.17±5.06) ％,CD86 + (97.74+0.87) ％.Proliferation index of PBLs primed by DCs three times was 15.4± 1.5.Cytokine levels including IL-2,IL-12,IFN-γ and TNF-α were obviously higher than nonpriming PBLs[(2551.9+195.3) pg/ml,(554.9±64.0) pg/ml,(2416.9±281.7) pg/ml,(632.4 +71.1)pg/ml,respectively] (P＜0.05),but IL-10 was no significant difference between priming CTLs and nonpriming CTLs.The average percentage of special CTLs was obviously higher than control group[(6.32±1.54)％,P＜0.05].The killing effect of CTLs was obviously higher than control group(P＜0.05).Conclusion Dendritic cells pulsed with peptide E613-21 and E786-94 can induce special CTLs in vitro and stimulate CTLs secret cytokines.This will provide science basis for research of therapeutic HPV vaccine.

Objective To study the impact of hepatitis B virus (HBV) infection on the activity of cord hematopoietic stem cells. Methods CD34+ cells were isolated from healthy human cord blood by miniMACS. Cells were cultured in IMDM complete culture medium containing stem cell factor (SCF),fms-like tyrosine kinase 3 ligand (FL), thrombopoietin (TPO), interleukin-3 (IL-3) and 10% fetal bovine serum. High copies HBV were added to the culture system. The proliferation of stem ceils and virus replication were observed. Following the proliferation, dendritic cells (DCs) were induced by adding granulocyte-macrophage colony-stimulating factor and IL-4. Morphous of stem cells and DCs were observed by microscope and the cell surface molecules were detected. Results The proliferation of stem cells infected with HBV was significantly lower than that of healthy stem cells (P＜0.01),and enhanced after adding cytokines (P＜0.01). At the same time, HBV replication was increased after adding cytokines in the culture system (P＜0.01), but the proliferation was still lower than that of healthy stem cells with cytokines in the culture medium (P＜0.05). Dane particles were found in the cytoplasma of stem cells infected with HBV by electron microscope. The expression of CD80,CD86 ,CD1a and HLA-DR on DCs derived from HBV infected stem cells were all lower than those on DCs from non-infected stem cells (P＜0.01). Conclusions HBV could infect CD34+ stem cell and the proliferation of the stem cell could enhance the virus replication. HBV could not only inhibit the proliferation of stem cells,but also down-regulate the immuno-phenotype expression of DCs derived from CD34+stem cells.

Objective To analyze the influence of the mean energy and the full-width of half msximum(FWHM)of incident electron beam intensity distilbution(assumed Gaussian distribution)on depth dose curves and off-axis ratios and to derive a most optimal combination of mean energy and FWHM of incident electron beam intensity distribution.Methods The study simulated 6 MV photon beam produced by Varian 600C medical linear accelerator with OMEGA/EGSnrc by matching the relative error of calculated and measured depth dose curves past depth of maximum dose and off-axis ratios at a depth of 10.0 cm in water within 2%.Results The depth dose curves were relatively insensitive to the mean energy past depth of maximum dose and the FWHM of the incident electron beam intensity distribution.Dose profiles were sensitive tO the mean energy and FWHM.The dose profiles horns decreased as the mean energy and tlle FWHM of the ineident electron beam intensity distilbution increased.The calculated value of the depth dose curves matched well with the measured value.The calculated value of the off-axis ratio was consistent with the measured value within the radiation field.However, the maximum errors of individual measurement points in the penumbra region and OUt of the field reached 18.5%.Conclusions In the field.the most optimal combination of mean energy and FWHM of incident electron beam intensitv distribution Can be derived, however,can not be derived out of the field and in the penumbra region.

Objective To study the topical application of Tuhuai decoction and its disassembled prescriptions on skin function of mice with epidermal hyperplasia, and to provide a theoretical basis for the treatment of psoriasis. Methods According to the Denda method, skin barrier function in mice was destructed with tape, to produce epidermal hyperplasia model. Tuhuai decoction and its disassembled prescriptions with different concentrations of water and alcohol extracts were external applied on the mice. Since the beginning of the treatment, loss of skin moisture of both sides the trunk was detected with Multi-functional instrument at every day before using the medicine. Results Tuhuai decoction had function of reducing the amount of water loss of skin. The disassembled prescriptions, such as Angelica group, safflower group, Danshen root group, and water and ethanol extracts of Prtmella group can reduce the amount of water loss of skin effect. Conclusion Water and ethanol extracts of Tuhuai decoction has the functions of reducing water loss of skin and resisting hyperplasia,suitable for treating psoriasis by external application, setting a foundation for clinical treatment of psoriasis.

BACKGROUND:Poststroke depression is one of the most common psychological behavior disorders after stroke and its mechanism remains unclear. Studies have suggested that microRNAs (miRNAs) involved in neurogenesis and synaptogenesis may play an important role in psychology diseases. OBJECTIVE:To observe the expression of miR-137 in the blood and brain of poststroke depression rats and its effect on the behaviors of rats. METHODS:Thirty-six rats were equal y divided into six groups:control, model, agomir-137, agomir-NC, agomir-137+Grin2A and agomir-137+vector groups. Control group had no treatment. Poststroke depression models were established by ligation of middle cerebral artery and chronic mild stimulation in the latter four groups fol owed by receiving an injection of nothing, agomir-137, agomir-NC, LV-CMV-Grin2A or control plasmids into the left lateral ventricle, respectively. RESULTS AND CONCLUSION:We found significantly lower miR-137 levels in the brain and peripheral blood of post-stroke depression rats compared with normal rats. Vertical scores and horizontal scores on the behavior test were significantly higher in the agomir-137 group than the agomir-NC and model groups at 3 weeks after cerebral ischemia;while, sucrose consumption percentage was also higher in the agomir-137 group at the end of 2 weeks after cerebral ischemia. Luciferase assays showed miR-137 bound to the 3’ UTR of Grin2A, regulating Grin2A expression in a neuronal cel line. Grin2A gene overexpression in the brain of post-stroke depression rats noticeably suppressed the inhibitory effect of miR-137 on post-stroke depression. Overal , these findings show that miR-137 suppresses Grin2A protein expression through binding to Grin2A mRNA, thereby exerting an inhibitory effect on post-stroke depression and offering a new therapeutic target for poststroke depression.

Objective To analyze the papers published by domestic scientific research workers in order to improve the academic level of their papers. Methods The papers published by over 3000 domestic scientific research workers were investigated with questionnaires. Their motives to publish papers and the relation between the number of pub-lished papers and the assessment of their performance were analyzed. Results The number of papers published by domestic scientific research workers was increased. However, their academic level was to be further improved. Over quantization of the assessment mechanisms for scientific research increased the external motives to publish papers, thus leading to the insufficient internal motives of them to engage in scientific research. Conclusion A loose and comfortable academic environment should be created for the scientific research workers in order to initiate their in-ternal motives to publish papers. Over quantization of the assessment mechanisms for scientific research should be changed in order to reduce the external motives of domestic scientific research workers to publish papers. Innovative and cultural environment should be created in order to improve the soft power of scientific research in our country.