Clinical significance of high mobility group protein B1 in patients with severe pneumonia complicated with sepsis / 新乡医学院学报

Objective To investigate the clinical significance of the changes of serum level of high mobility group protein B1 (HMGB1) in patients with severe pneumonia complicated with sepsis.Methods Fifty patients with severe pneumonia complicated with sepsisin in Respiratory Intensive Care Unit(RICU) of Xinxiang Central Hospital from April 2014 to March 2017 were selected as observation group;while 50 healthy individuals were selected as control group.The patients in the observation group were divided into death group(n =32) and survival group(n =18) according to the prognosis.The serum levels of procalcitonin(PCT) and HMGB1 of patients in the observation group were detected on the 1st,3rd,7th day of patients hospitalized in the RICU,while the acute physiology and chronic health evaluation Ⅱ (APACHE lⅡ)scores of the patients were evaluated.The serum levels of PCT and HMGB1 of subjects in the control group were detected during physical examination.Results There was no statistic difference in the mean arterial pressure,oxygenation index,body temperature and total white cell count of patients between the death group and survival group(P ＞0.05).On the first day of patients hospitalized in the RICU,the serum levels of PCT and HMGB1 of patients in the observation group were significantly higher than those in the control group (P ＜0.05).The serum levels of HMGB1 and the APACHEⅡ scores of patients in the death group were significantly higher than those in the survival group at each time point(P ＜0.05).On the first day of patients hospitalized in the RICU,there was no statistic difference in the serum level of PCT of patients between the death group and survival group (P ＞ 0.05);the serum level of PCT of patients in the death group was significantly higher than that in the survival group at another time point (P ＜ 0.05).The serum level of HMGB1 of patients in the observation group was positively correlated with the PCT and APACHE Ⅱ score (r =0.562,0.460;P ＜0.05).Conclusion The serum level of HMGB1 in patients with severe pneumonia complicated with sepsis is increased;and the increase of serum level of HMGB1 in the death cases is more obvious than that in the survival cases.So it can be used to evaluate the patient's condition and judge the prognosis.

Clinical observation of exenatide for treatment of obesity type 2 diabetes mellitus complicated with obstructive sleep apnea hypopnea syndrome / 新乡医学院学报

Objective To observe the clinical effect of exenatide for treatment of obesity type 2 diabetes mellitus (T2DM) conplicated with obstructive sleep apnea hypopnea syndrome(OSAHS).Methods Eighty patients of obesity T2DM complicated with OSAHS in the Department of Endocrinology,the First People's Hospital of Xinxiang from August 2014 to August 2016 were chosen and randomly divided into observation group(n =40) and control group(n =40).The patients in control group taken orally mefformin 800 mg twice daily for 24 weeks;based on this,the patients in observation group were injected subcutaneously exenatide 10 μg at one hour before meal,twice daily for 24 weeks.The fasting blood glucose (FBG),2-hour postprandial blood glucose (2 hPBG),glycosylated hemoglobin (HbA1 C),body mass index (BMI),waistline,total cholesterol (TC),triglyceride (TG),apnea hypoventilation index (AHI),Epworth sleepiness scale (ESS) score,the lowest oxygen saturation in sleep (LSpO2) were compared between the two groups before and after treatment.Results There was no statistic difference in the HbA1 C,FPG,2 hPBG levels of patients between the two groups before treatment (P ＞ 0.05);the HbA1 C,FPG,2 hPBG levels of patients in the two groups after treatment were significantly lower than those before treatment (P ＜ 0.05);and the HbA1 C,FPG,2 hPBG levels of patients in observation group were significantly lower than those in the control group after treatment(P ＜ 0.05).There was no statistic difference in the BMI,waistline and TC,TG levels of patients between the two groups before treatment(P ＞ 0.05);the BMI,waistline and TC,TG levels of patients in the two groups after treatment were significantly lower than those before treatment (P ＜ 0.05);and the BMI,waistline and TC,TG levels of patients in observation group were significantly lower than those in the control group after treatment(P ＜ 0.05).There was no statistic difference in the AHI,ESS scores,SpO2 and LSpO2 levels of patients between the two groups before treatment(P ＞0.05);the AHI,ESS scores,SpO2 and LSpO2 levels of patients in the two groups after treatment were significantly lower than those before treatment(P ＜0.05);and the AHI,ESS scores,SpO2 and LSPO2 levels of patients in observation group were significantly lower than those in the control group after treatment(P ＜0.05).The incidence of adverse reaction of patients in observation group and control group was 7.5％ (3/40) and 5.0％ (2/40) respectively;there was no statistic difference in the incidence of adverse reaction between the two groups (P ＞ 0.05).Conclusion Exenatide can effectively control blood glucose,improve the function of beta cells and reduce insulin resistance.It can effectively reduce body weight,BMI and waistline,improve the quality of sleep breathing.

1,3,4-tri-O-galloyl-6-O-caffeoyl-β-D-glucopyranose, a new anti-proliferative ellagitannin, regulates the expression of microRNAs in HepG(2) cancer cells / 南方医科大学学报

<p><b>OBJECTIVE</b>MicroRNAs (miRNAs) play important roles in cell proliferation, differentiation and apoptosis. 1, 3, 4-tri-O-galloyl-6-O-caffeoyl-β-D-glucopyranose (BJA32515) is a new natural ellagitannin compound extracted from Balanophora Japonica MAKINO. The effect of BJA32515 on the expression of miRNAs in cancer cells has not yet been explored. Objective The present study was carried out to examine the changes in miRNA expression profiles in human HepG(2) hepatocarcinoma cells following BJA32515 exposure.</p><p><b>METHODS</b>The proliferation of BJA32515-exposed HepG(2) cells was assessed using a colorimetric assay (cell counting kit-8). The miRNA expression profile of the cancer cells was analyzed using a miRNA array and quantitative real-time PCR. Apoptosis was assessed by annexin V and propidium iodide staining.</p><p><b>RESULTS</b>BJA32515 inhibited the cell proliferation and increased apoptosis in HepG(2) cancer cells. The exposure to BJA32515 also caused alterations in the miRNA expression profile in the cells, with 33 miRNAs upregulated and 59 down-regulated. The up-regulation of let-7a and miR-29a and the down-regulation of miR-373 and miR-197 were verified by quantitative real-time PCR. CONCLSION: BJA32515-modifed miRNA expression may mediate the antiproliferative effect of this compound in HepG(2) cancer cells.</p>

Synthesis and antitumor activities of pyrimidines / 南方医科大学学报

<p><b>OBJECTIVE</b>To synthesize cyclin-dependent kinase (CDKs) inhibitors and assay their antitumor activities.</p><p><b>METHODS</b>A series of pyrimidines containing different arylamino and 1-(methylsulfonyl)piperidin moieties were designed by combining the segments 1-(methylsulfonyl)piperidin and pyrimidine heterocycles according to the super-position principle of the reinforcement of biological activities.</p><p><b>RESULTS</b>Their structures were characterized by MS and 1H NMR spectra and all the synthesized compounds were screened for their antimicrobial activity with MTT assay.</p><p><b>CONCLUSION</b>The preliminary bioassay showed that compound 3 b displayed good antitumor activity (IC(50)=13.6 µmol/L). The preliminary structure activity relationship analysis of these analogues suggest that the steric factor may have important impact on the anti-tumor activity.</p>

Construction of rheumatoid arthritis-specific full-length fully human mammalian display antibody libraries / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a rheumatoid arthritis-specific full-length fully human mammalian display antibody libraries.</p><p><b>METHODS</b>Peripheral blood lymphocytes were isolated from patients with rheumatoid arthritis. The repertoires of kappa light chain (LCκ) and heavy chain variable region (VH) of the antibodies were amplified by RT-PCR. The amplified LCκ and VH genes were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO-10 strain to construct the rheumatoid arthritis-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length fully human antibody expressed on the surface of 293T cells were analyzed by flow cytometry.</p><p><b>RESULTS</b>The libraries of rheumatoid arthritis-specific antibody LCκ and heavy chain (IgG1) were constructed. The expression of full-length fully human antibody on the surface of 293T cells was confirmed by flow cytometry. With the rates of correct LCκ and heavy chain sequence insertion reaching 80% and 60%, respectively, as shown by DNA sequence analysis of the randomly selected clones, the libraries showed an expressible combinatory diversity of 6.13×10(10).</p><p><b>CONCLUSION</b>The constructed libraries provide a useful platform for screening rheumatoid arthritis-specific antibodies.</p>

Cough-relieving, analgesic and antibiotic effects of durian shell extracts: a study in mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the cough-relieving, analgesic and antibiotic effects of durian shell extract (DSE) in relieving cough and its analgesic and antibiotic effects.</p><p><b>METHODS</b>The effect of DSE in relieving cough was assessed in mice challenged with ammonia and SO(2) to induce coughing. The analgesic and antibiotic effects of DSE in mice were evaluated by hot plate test and twisting reaction induced by acetic acid, and by minimal inhibitory concentration (MIC) and disc-agar diffusion tests, respectively.</p><p><b>RESULTS</b>Compared with the control group, the mice treated with 300 and 900 mg/kg DSE showed significantly prolonged latency with decreased number of coughing induced by ammonia and SO(2), and the effect was dose-dependent. DSE markedly prolonged the latency and decreased the twisting number of the mice induced by acetic acid without affecting the pain threshold in hot plate test. DSE produced no significant inhibitory effects against Staphylococcus aureus, Staphylococcus epidermidis, or E. coli, and showed a week inhibition against Bacillus aeruginosus.</p><p><b>CONCLUSION</b>DSE shows obvious effect in relieving cough and produces better analgesic effect against chemical factor-induced pain than against physical agent-induced pain sensation. DSE has a moderate inhibitory effect against Bacillus aeruginosus.</p>

Naringin inhibits monocyte adhesion to high glucose-induced human umbilical vein endothelial cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the inhibitory effect of naringin on monocyte adhesion to high glucose-induced human umbilical vein endothelial cells (HUVECs).</p><p><b>METHODS</b>Cultured HUVECs isolated from human umbilical cords were pretreated with or without naringin and induced with high glucose (33 mmol/L) for 48 h. Human monocyte THP-1 cells, after labeling with BCECF-AM, were co-cultured with the HUVECs for 30 min. The labeled THP-1 cells adhering to HUVECs were observed under fluoroscence microscope, and the inhibitory effect of naringin on the cell adhesion was evaluated by measuring the adhering cell density. Western blot analysis was used to detect the expressions of the adhesion molecules in the HUVECs, and reactive oxygen species (ROS) production in the HUVECs was measured using an oxidation-sensitive fluorescent probe (DCFH-DA). The nuclear extracts of the HUVECs were prepared to examine the expression of nuclear factor-kappa B (NF-kappaB) in the cell nuclei by Western blotting.</p><p><b>RESULTS</b>HUVECs in high-glucose culture showed increased adhesion to THP-1 cells and enhanced expressions of the cell adhesion molecules, which were significantly attenuated by pretreatment with naringin (10-50 microg/ml). High glucose induced DCF-sensitive intracellular ROS production in the HUVECs, and this effect was inhibited by naringin pretreatment of the cells. Naringin also suppressed high glucose-induced increment of NF-kappaB expression in the cell nuclei of HUVECs.</p><p><b>CONCLUSION</b>Naringin can suppress high glucose-induced vascular inflammation possibly by inhibiting ROS production and NF-kappaB activation in HUVECs.</p>

Immunomodulatory effects of Fomes fomentarius polysaccharides: an experimental study in mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the immunomodulatory effects of Fomes fomentarius polysaccharides (FFP) in mice.</p><p><b>METHODS</b>MTT assay was employed to evaluate the in vitro metabolic activity of the mouse splenocytes treated with FFP at different concentrations, and the secretion of tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (INF-gamma) and interleukin 2 (IL-2) from the cells were measured by enzyme-linked immunosorbent assay. The changes in the phagocytotic activity of mouse macrophage in response to FFP treatment were evaluated by phagocytosis percentage of chicken red blood cells (CRBCs). The effect of FFP on the humoral immunity was assessed in mice immunized with sheep red blood cells (SRBCs) by measuring the serum levels of specific antibody (hemolysin) against SRBCs.</p><p><b>RESULTS</b>FFP at the concentrations of 25, 50, and 100 microg/ml all significantly enhanced the metabolic activity of mouse splenocytes in vitro and increased the production of TNF-alpha, IFN-gamma and IL-2. FFP treatment also markedly enhanced the metabolic activity of mouse peritoneal exudate cells and TNF-alpha production by the cells. At the doses of 25, 50, and 100 mg/kg, FFP significantly increased serum hemolysin level in mice immunized with SRBCs, and FFP at 50 and 100 mg/kg obviously increased the capacity of mouse peritoneal macrophages in vivo for CRBC phagocytosis.</p><p><b>CONCLUSION</b>FFP can promote the secretion of TNF-alpha, IFN-gamma and IL-2 by mouse immunocytes and enhance mouse humoral immune response and the phagocytotic activity of the macrophages.</p>

Construction of a bait plasmid containing human telomerase RNA mutant in yeast three-hybrid system / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a bait plasmid containing human telomerase RNA with multiple point mutations in a yeast three-hybrid system and evaluate the toxicity of the recombinant bait plasmid.</p><p><b>METHODS</b>The primers were designed according to the hTR sequence and the target mutation sites for inducing T-->A mutations at the 41st, the 80th and 102nd nucleotides of the hTR gene using the overlapping extension PCR (OE-PCR) method. The mutant was cloned into PMD18T vector, confirmed by sequencing, sub-cloned into the bait plasmid PRH3' and identified with PCR and restriction enzyme digestion. The recombinant bait plasmid was then transformed into yeast L40 ura3/pHyblex/ZeoMS2 for toxicity test.</p><p><b>RESULTS</b>Sequence analysis demonstrated successful introduction of point mutations at the target sites without causing random mutation. The recombined bait plasmid constructed showed no obvious toxicity against the host yeast cells.</p><p><b>CONCLUSIONS</b>The recombinant plasmid containing the human telomerase RNA mutant (PRH3'-hTRm) has been successfully constructed and can be used as the bait plasmid in yeast three-hybrid system.</p>

Establishment of mouse model of humoral immune response using rabbit red blood cells as the antigen / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a mouse model of humoral immune response by immunization with rabbit red blood cells (RRBCs).</p><p><b>METHODS</b>The mice were immunized with RRBCs and the serum hemolysin level was measured by micro-hemolysis spectrophotometry.</p><p><b>RESULTS</b>The peak time needed for hemolysin production against RRBCs was 6 days after the immunization, and 20% RRBCs in a total volume of 0.2 ml was optimal for intraperitoneal injection. Hydrocortisone (25 mg/kg) and cyclophosphamide (20 mg/kg) inhibited hemolysin production. Mannatide (4 mg/kg) produced no significant effect on serum hemolysin level in normal mice, but significantly potentiated hemolysin production in immunosuppressed mice induced by cyclophosphamide (20 mg/kg).</p><p><b>CONCLUSION</b>Intraperitoneal RRBC injection is feasible for establishing mouse models of humoral immune response.</p>

Establishment of homogeneous time-resolved fluorescence immunoassay for high throughput screening of protein tyrosine kinase inhibitors / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish an in vitro homogeneous time-resolved fluorescence immunoassay method for high throughput screening of protein tyrosine kinase (PTK) inhibitors.</p><p><b>METHODS</b>Specific fluorescence signals at 670 and 612 nm were measured by multifunctional microplate reader when the fluorescence was emitted through a resonance energy transfer between fluorescent materials (EuK and XL-665). The inhibitory activity of Sunitinib, a standard PTK inhibitor, on vascular endothelia growth factor receptor 2 (VEGFR-2) kinase activity was investigated.</p><p><b>RESULTS</b>A homogeneous time-resolved fluorescence immunoassay was established for high throughput screening of PTK inhibitor. In this system, the concentrations of VEGFR-2, adenosine triphosphate (ATP) and poly-peptide substrate were 5 ng/microl, 100 micromol/L and 1 micromol/L, respectively. Sunitinib inhibited VEGFR-2 kinase activity with an IC50 value of 86.7 nmol/L, which was close to the values tested using other methods.</p><p><b>CONCLUSION</b>The homogeneous time-resolved fluorescence immunoassay we established can be easily used for high throughput screening of PTK inhibitors.</p>

Hydrocamptothecin promotes the mRNA expression of Wnt signaling inhibitor DKK-1 / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of hydrocamptothecin on the expression of Wnt signaling pathway inhibitor DKK-1 in tumor cells.</p><p><b>METHODS</b>Human HepG2, Hep3B, LoVo and U251 cells were treated with the antitumor drug Hydrocamptothecin. DKK-1 mRNA expression in the cells was detected with RT-PCR, and beta-catenin expression was measured by fluorescence-activated cell sorting (FACS).</p><p><b>RESULTS</b>DKK mRNA in Hep3B, HepG2, LoVo and U251 cells was significantly increased after hydrocamptothecin treatment for 24 h, and the percentage of beta-catenin-positive cells and fluorescence intensity for beta-catenin expression was lowered in the cells after the treatment.</p><p><b>CONCLUSION</b>Hydrocamptothecin promotes mRNA expression of Wnt signaling pathway inhibitor DKK-1 in Hep3B, HepG2, LoVo and U251 cells.</p>

1,2,6-tri-O-galloyl-beta-D-glucopyranose inhibits gp41-mediated HIV envelope fusion with target cell membrane / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the inhibitory effect of 1,2,6-Tri-O-galloyl-beta-D-glucopyranose (TGGP) from Balanophora japonica Makino on human immunodeficiency virus (HIV) entry into the host cells and explore the mechanisms.</p><p><b>METHODS</b>TGGP was purified from Balanophora japonica Makino by n-hexane and ethyl acetate extraction and column chromatography. The inhibitory activity of TGGP on HIV gp41 six-helix bundle formation was measured with ELISA, N-PAGE and SE-HPLC, and the inhibitory effect of TGGP on HIV envelope grlycoprotein-induced cell-cell fusion was detected using a non-infectious cell-based assay.</p><p><b>RESULTS</b>TGGP inhibited HIV gp41 six-helix bundle formation, with an IC50 of 1.37-/+0.19 microg/ml as determined by ELISA, and this activity was further confirmed by N-PAGE and SE-HPLC. TGGP at 25 microg/ml significantly inhibited syncytium formation between the effector (CHO-WT) and the target (MT-2) cells.</p><p><b>CONCLUSION</b>The HIV transmembrane subunit gp41 mediates the entry of HIV into the target cells. TGGP can inhibit HIV fusion and entry into the target cells by inhibiting the formation of gp41 six-helix bundles, suggesting the potential of TGGP as a microbicide to prevent sexual transmission of HIV.</p>

Immunological activity of Streptomyces polysaccharide in normal and immunosuppressed mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the immunological activity of Streptomyces polysaccharide (SMP) on normal and immunosuppressed mice.</p><p><b>METHODS</b>The effect of SMP on the proliferating activity of normal mouse splenocytes was tested in the mixed lymphocyte culture, and the changes of peripheral blood T lymphocytes were evaluated with acid a-naphthyl acetate esterase (ANAE) method. The ratio of Lyt2+ and L3T4+ T cell subsets was measured by flow cytometry.</p><p><b>RESULTS</b>SMP stimulated obvious proliferation of mixed lymphocytes, showed protective effects on T lymphocyte and increased the ratio of Lyt2+ and L3T4+ cell subsets to nearly normal level in immunosuppressed mice.</p><p><b>CONCLUSIONS</b>SMP can regulate the immune function in mice.</p>

Saponin from Tupistra chinensis Baker inhibits mouse sarcoma S-180 cell proliferation in vitro and implanted solid tumor growth in mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the antitumor effect of saponin extracted from Tupistra chinensis Baker (STCB) against mouse sarcoma S-180 cell proliferation in vitro and in vivo and explore the primary mechanism of this effect.</p><p><b>METHODS</b>Cytotoxic effect of STCB on S-180 cells in vitro was evaluated by MTT colorimetry, and its effect against in vitro tumor growth was tested in Kunmin mice bearing S-180 implanted tumor. The morphological and ultrastructural changes of S-180 cells after saponin treatment in vitro were examined with light and transmission electron microscope. Flow cytometry was performed to examine the cell cycle and apoptosis of S180 cells treated with different concentrations of STCB with propidium iodide staining.</p><p><b>RESULTS</b>STCB could markedly inhibit S-180 cell proliferation in vitro with 50% inhibitory concentration of 34.64 microg/ml. STCB given by intragastric administration also significantly inhibited the growth of S-180 solid tumor, and the inhibition rate exceeded 30% at the dose of 0.5 g/kg, reaching 54.86% at 2 g/kg. Electron microscopy and flow cytometry revealed increased S180 tumor cell apoptotic rate with the increment of saponin concentration, along with increased percentage of cells in S phase and decreased cells in G(2)/M phase in response to 10 or 30 microg/ml STCB treatment. At the concentration of 60 microg/ml, however, STCB resulted in an opposite effect on the cell cycles, presumably due to its interference with mitosis at high concentrations.</p><p><b>CONCLUSIONS</b>STCB inhibits the growth of S-180 cells both in vivo and in vitro possibly by inducing cell apoptosis and interfering with the cell cycle progression of the tumor cells.</p>

Inhibitory effect of polysaccharides on the six-alpha-helix bundle formation of HIV gp41 protein / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare the in vitro inhibitory effect of expolysaccharides from Streptomyces, polysaccharides of Ganoderma lucidum and rice bran on six-alpha-helix bundle formation of HIV gp41 protein.</p><p><b>METHODS</b>The amount of six-alpha-helix bundle formed in the presence of N36 and C34 was tested by ELISA in response to treatments with different doses of polysaccharides.</p><p><b>RESULTS</b>Expolysaccharides from Streptomyces potentially inhibited six-alpha-helix bundle formation with the effective concentration (IC(50)) of 145.48-/+7.25 mg /L. Polysaccharides of Ganoderma lucidum and rice bran showed no effect on the six-alpha-helix bundle formation.</p><p><b>CONCLUSION</b>Expolysaccharides from Streptomyces can inhibit the six-alpha-helix bundle formation of HIV gp41, whereas polysaccharides of Ganoderma lucidum and rice bran do not exhibit such activity.</p>

Effect of Ganoderma lucidum polysaccharides on tumor cell nucleotide content and cell cycle in S180 ascitic tumor-bearing mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of Ganoderma lucidum polysaccharides (GLP) on the nucleotide contents and cell cycle distribution of the tumor cells in S180 ascitic tumor-bearing mice and explore the possible mechanism of the antitumor effect of GLP.</p><p><b>METHODS</b>Mice bearing S180 ascitic tumor were subjected to intragastric administration of GLP (100, 200, and 400 mg/kg), normal saline or subcutaneous injection of cyclophosphamide (CTX) at 25 mg/kg, respectively. The treatment was given once daily for 9 consecutive days, after which the ascitic tumor cells were harvested for determination of the RNA and DNA contents and their ratio as well as the cell cycle alterations. Laser scanning confocal microscopy and acridine orange staining was performed to evaluate the DNA and RNA fluorescence intensity, and flow cytometry with propidium iodide (PI) staining was utilized for cell cycle analysis of the tumor cells.</p><p><b>RESULTS</b>Compared with normal saline group, the tumor cells in the 3 GLP groups all showed reduced RNA and DNA contents, and this reduction was statistically significant in 200 mg/kg GLP group (P=0.000). Significantly reduced RNA/DNA ratio was noted in all the 3 GLP groups (P=0.003, 0.000, 0.008 corresponding to 400, 200, and 100 mg/kg groups), suggesting that ganoderma polysaccharides more effectively reduced RNA content than DNA content. CTX also resulted in reduced RNA and DNA contents but not the RNA/DNA ratio. At the doses of 400, 200, and 100 mg/kg, GLP increased the percentage of G2/G2 phase cells (P=0.003, 0.000, and 0.000) whereas CTX showed the contrary effect (P=0.000). GLP produced no obvious effect on S-phage cells but CTX significantly reduced their percentage (P=0.000). GLP at the 3 doses all decreased the percentage of G2/M phase tumor cells (P=0.014, 0.049, 0.016) and CTX again induced contrary effect (P=0.000).</p><p><b>CONCLUSION</b>With different effects from CTX on DNA and RNA contents and cell cycle, GLP inhibits DNA and RNA synthesis in the tumor cells by mobilizing the host immune function to interfere with the normal cell cycles, which might be one of the mechanisms for the antitumor effect of GLP.</p>

Purification of a new phospholipase A2 homologue from Agkistrodon blomhoffii siniticus and its effects on gene expression profile of Hep3B cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To isolate and purify a new phospholipase A2 (PLA2) homologue from Agkistrodon blomhoffii siniticus and investigate its effects on the gene expression profile of Hep3B cells.</p><p><b>METHODS</b>The PLA2 homologue was isolated and purified by reverse-phase high-performance liquid chromatography (HPLC) and its purity was determined also by HPLC. The relative molecular mass of the homologue was measured by electrospray ionization mass spectrum. The gene expression profile of Hep3B cells was detected with gene chip after exposure of the cells to 139 microg/ml PLA2 homologue for 12 h.</p><p><b>RESULTS</b>The purity of the PLA2 homologue was 97.2%, whose relative molecular mass was 13,900. After exposure of Hep3B cells to 139 microg/ml PLA2 homologue for 12 h, 19 genes were down-regulated and 20 up-regulated in the cells. The genes showing altered expressions in response to the exposure were mainly involved in cell cycle control and DNA damage repair, cell apoptosis and senescence, production of signal transduction molecules and transcription factors, cell adhesion, angiogenesis, and tumor invasion and metastasis.</p><p><b>CONCLUSIONS</b>The PLA2 homologue induces alterations in the expression of a wide variety of genes involved in the growth and metastasis of tumor cells. The results of this study provide clues for further study of the possible mechanism for the action of PLA2 homologue on Hep3B cells.</p>

Ganoderma polysaccharides antagonize prostaglandin E2-induced suppression of murine splenocyte IFN-gamma and TNF-alpha mRNA expression / 南方医科大学学报

<p><b>OBJECTIVE</b>To determine if Ganoderma polysaccharides can antagonize prostaglandin E2 (PGE2)-induced suppression of murine splenocyte interferongamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) mRNA expression.</p><p><b>METHODS</b>Mixed lymphocyte culture reaction was used as the experimental model. The expressions levels of IFN-gamma and TNF-alpha mRNA were measured by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>After the cultures were treated with PGE2 for 4 h, IFN-gamma mRNA expression was reduced as compared with the control, which was especially obvious when PGE2 concentrations exceeded 10 micromol/L (P<0.01). Ganoderma polysaccharides above 100 mg/L showed partial antagonistic effect against the inhibition of IFN-gamma by PGE2 at the fixed concentration of 20 micromol/L. Further studies indicated that PGE2 (20 micromol/L) impaired the expression of TNF-alpha mRNA after an 8-hour incubation and Ganoderma polysaccharides above 100 mg/L could partially antagonize this effect.</p><p><b>CONCLUSION</b>Ganoderma polysaccharides can partially antagonize PGE2-induced suppression of murine splenocyte IFN-gamma and TNF-alpha mRNA expression.</p>

Diclofenac suppresses hepatoma cell proliferation and promotes cyclooxygenase-2 mRNA expression / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of cyclooxygenase inhibitor diclofenac on the proliferation and cyclooxygenase-2 (COX-2) mRNA expression of cultured hepatocellular carcinoma cell lines HepG2, Hep3B and human hepatocellular cell line QSG-7701.</p><p><b>METHODS</b>After exposure to diclofenac at various concentrations (10-200 micromol/L) for 24, 48 and 72 h, the cell proliferation was analyzed by Cell Counting Kit-8 (CCK-8) assay and mRNA expression determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>Diclofenac exposure for 24, 48 and 72 h significantly inhibited HepG2 and Hep3B cell proliferation in a concentration-dependent manner, with inhibition rate of 40.47% and 54.49% after 48 h exposure to 50 micromol/L diclofenac and IC50 of 70.54 and 48.39 micromol/L, respectively. A much weaker antiproliferative effect on QSG-7701 cells was shown, with IC50 of 189.91 micromol/L after 48-hour exposure to diclofenac. RT-PCR detected COX-2 mRNA in HepG2 and Hep3B cells, but hardly in QSG-7701 cells. Treatment with diclofenac or 5-Fu resulted in elevated COX-2 mRNA expression both in HepG2 and Hep3B cells.</p><p><b>CONCLUSION</b>Diclofenac can specifically inhibit the proliferation of COX-2-expressing HepG2 and Hep3B cells, and induce up-regulation of COX-2 mRNA expression, which indicates the important role of COX-2 in the proliferation of hepatoma cells.</p>