OBJECTIVEThe study aimed to test if Paridis Rhizoma total saponins (PRTS) could induce apoptosis of human gastric cancer cell MKN-45.

METHODBased on the previous researches, PRTS was set by different concentrations to treat human gastric cancer cell for 12 h (5, 10, 20 mg x L(-1)). Fluorescent staining methods were adopted to observe apoptotic morphological changes of MKN-45. The apoptosis rates were analyzed by flow cytometry with Annexin V-FITC/PI staining. The enzymatic activities of caspase-3 and caspase-8 were measured by ELISA. The protein levels of Fas and FasL were detected by Western blotting.

RESULTUnder a fluorescence microscope, MKN-45 treated by PRTS was seen typical apoptotic morphological features. PRTS significantly increased the rate of apoptosis. Compared with the control group, there exsited significant differences in apoptosis rate of PRTS concentration of 20 mg x L(-1) (P < 0.01); besides, the enzymatic activities of caspase-3 and caspase-8 were promoted obviously after the effect of PRTS on MKN-45 cells for 12 h (P < 0.01). The protein levels of Fas and FasL in the MKN-45 were upgraded significantly.

CONCLUSIONPRTS can induce apoptosis of human gastric cancer cell MKN-45 , which is concerned with caspase-3 and caspase-8 and upgraded Fas and FasL.

Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Caspase 8 ; genetics ; metabolism ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; Fas Ligand Protein ; metabolism ; Humans ; Magnoliopsida ; chemistry ; Rhizome ; chemistry ; Saponins ; pharmacology ; Signal Transduction ; drug effects ; Stomach Neoplasms ; drug therapy ; genetics ; metabolism ; physiopathology ; fas Receptor ; metabolism

Animals ; Dietary Carbohydrates ; administration & dosage ; Dietary Fats ; administration & dosage ; Dietary Proteins ; administration & dosage ; Humans ; Malnutrition ; complications ; Obesity ; etiology ; Trace Elements ; administration & dosage

Teaching method for basal experiment, comprehensive experiment, design experiment and teach- ing practice in food microiological analysis were elaborated completely, and design experimental teaching was discussed stress. At the same time, Through introducing various experience of the design experiment teaching, resolvent and way of thinking against problem meeted in design experiment teaching were put forward.

Objective To summarize the preliminary experience of endovascular stent-graft exclusion for aortic dissections. Methods From October 2003 to February 2007,121 patients[86 males,37 females,mean age(53.7?13.8)years,range 29～ 72 years]underwent endovascular stent-graft exclusion for aortic dissections,including Stanford B in 114 patients,Stanford A in 4, and traumatic aortic mptore in 3.An emergency operation was performed in 4 patients for acute aortic rapture.Results No primary conversion was needed.There was no postoperative death,no spinal cord iscbemic injury,or stent displacement or subclavian steal syndrome.Postoperative hospital stay time was(4.0?1.3)days.Complications included fever in 35 patients,type Ⅳ endoleak in 11,type Ⅰ endoleak in 1 and acute renal dysfunction in 1.Contusion Endovascular thoracic aorta repair is an effective,less inva- sire and safe surgery for patients with Stanford B or some Stanford A aortic dissection and traumatic aortic rupture.

Objective To explore the efficacy of Fushe Jiedu Decoction combined with red light on viper bites injury limb swelling and the effects on the inflammatory cytokines. Methods Totally 90 patients were divided into control group and experimental group by using random number table method, with 45 cases in each group. The wounds of the control group were sterilized and given anti-snake venom serum, antibiotics, and tetanus immunoglobulin and supplemented with energy to correct water and electrolyte disturbances. The experimental group was treated with Fushe Jiedu Decoction based on the treatment of control group,150 mL each time,orally,twice a day; Red light was applied at the site of the most obvious swelling of the injured limb, 20 minutes each time, twice a day. The treatment lasted for 6 d. The swelling of injured limbs, serum C-reactive protein (CRP), 5-hydroxytryptamine (5-HT), histamine, tumor necrosis factor-α (TNF-α), interleukin (IL)-6 levels before and after treatment in the two groups were observed. Results Compared with before treatment, the swelling of the limbs disappeared significantly in the experimental group at 3 d and 6 d and in the control group at 6 d, with statistical significance (P<0.01). Compared with the control group at the same time point, the swelling of the limbs in the treatment group was significantly better than that in the control group at 3 d and 6 d, with statistical significance (P<0.01). Compared with before treatment, the levels of CRP, 5-HT, TNF-α, and IL-6 were significantly lower in the two groups, with statistical significance (P<0.05). After treatment, CRP, 5-HT and histamine in the experimental group were significantly better than that in the control group(P<0.05).Conclusion Fushe Jiedu Decoction combined with red light has good efficacy for viper bites injury limb swelling, which can reduce inflammatory cytokines levels of patients.

The relationship between tyrosine phosphorylation (TP) and protein expression of insulin receptor (InsR) and insulin resistance (IR) in patients with gestational diabetes mellitus (GDM) was investigated. The InsR expression and TP in skeleton muscle tissue were determined by Western blotting and immunoprecipitation in women with GDM (GDM group, n=22), normal pregnant women (normal pregnancy group, n=22) and normal non-pregnant women (normal non-pregnant group, n=13). Fasting plasma glucose (FPG) and fasting insulin (FINS) were measured by oxidase assay and immunoradioassay. The results showed that the levels of FPG (5.61±0.78 mmol/L), FINS (15.42±5.13 mU/L) and Homeostasis model assessment-IR (HOMA-IR) (1.21±0.52) in GDM group were significantly higher than those in normal pregnancy group (4.43±0.46 mmol/L, 10.56±3.07 mU/L and 0.80±0.31 respectively) (P<0.01). The levels of FINS and HOMA-IR in normal pregnancy group were significantly higher than those in normal non-pregnant group (7.56±2.31 mU/L and 0.47±0.26 respectively) (P<0.01). There was no significant difference in the InsR expression level among the three groups (P>0.05). TP of InsR with insulin stimulation was significantly decreased in GDM group (0.20±0.05) as compared with normal pregnancy group (0.26±0.06) (P<0.01). TP of InsR with insulin stimulation in normal pregnancy group was lower than that in normal non-pregnant group (0.31±0.06) (P<0.01). TP of InsR with insulin stimulation was negatively related with HOMA-IR in GDM group (r=-0.525, P<0.01). There was no correlation between the protein expression of InsR and HOMA-IR in GDM group (r=-0.236, P>0.05). It was suggested that there is no significant correlation between the protein expression of InsR in skeletal muscle and IR in GDM, but changes in TP of InsR are associated with IR in GDM.

Aged ; Genital Neoplasms, Male ; pathology ; Humans ; Male ; Neurilemmoma ; pathology ; Scrotum

Yeast is a widely used host for recombinant protein expression. However, glycoproteins derived from yeast contain N-glycan of high mannose type and are usually hyperglycosylated. alpha-1,6-mannosyltransferases gene (och1) encodes the enzyme that initiates the first step of out-chain elongation of high mannose type N-glycan in yeast, which is different from that in human. So, a high efficient method to knockout target gene by two-step recombination was established and was used to delete och1. In the first recombinant, a plasmid with och1::ADE1 and ura3 gene was linearized in the downstream of och1 and inserted to the och1 site of P. pastoris genome, where the upstream and downstream of och1 were duplicated. In the second recombinant, the duplicated fragments of och1 were exchanged and the och1 deletion strains were selected on the plates containing 5-FOA, but no adenine. Then the och1 deletion strain was applied to express an human serum albumin (HSA) granulocyte-macrophage colony-stimulating factor (GM-CSF) chimera. Different with the hyperglycosylated HSA/GM-CSF chimera expressed in wild type P. pastoris, the chimera expressed in the och1 deletion strain, contained smaller N-glycan. The results suggested that the och1 mutant yeast may be more suitable for production of recombinant glycoproteins. And the och 1 deletion strain could be used for further re-engineering to produce complex human glycoproteins.
Chimera ; Gene Deletion ; Gene Knockout Techniques ; Granulocyte-Macrophage Colony-Stimulating Factor ; biosynthesis ; genetics ; Mannosyltransferases ; genetics ; Pichia ; enzymology ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Serum Albumin ; biosynthesis ; genetics

Blood Cell Count ; Bone Marrow Cells ; metabolism ; Child ; Cord Blood Stem Cell Transplantation ; Cytokines ; therapeutic use ; Histocompatibility Testing ; Humans ; Male ; Myelodysplastic Syndromes ; blood ; therapy ; Treatment Outcome

OBJECTIVETo investigate the effect of decreased expression of high mobility group Box-1 on the proliferation and apoptosis of HepG2 cells.

METHODSThree specific siRNAs of HMGB1 were designed and synthesized, and were transiently transfected into HepG2 cells by Lipofectamine 2000. The HMGB1 expression in HepG2 cells was detected by RT-PCR and Western blotting respectively. The proliferation activity in vitro was assessed by MTT assay. In situ apoptosis was evaluated by terminal deoxynucleotidyl transferase-deoxyuridine triphosphate nick end labeling (TUNEL) assay.

RESULTSAll of these specific HMGB1-siRNAs (1, 2, 3) efficiently and specifically inhibited the expression of the HMGB1 gene, and the levels of HMGB1 mRNA were 1.147+/-0.024, 1.014+/-0.042, 0.435+/-0.055, respectively, in HMGB1-siRNAs transfection group, which were significantly lower than that in Lipofectamine 2000 alone group (1.411+/-0.065, P < 0.01). Correspondingly, all of these specific HMGB1-siRNAs (1, 2, 3) could efficiently and specifically inhibit the expression of the HMGB1 protein, and the levels of HMGB1 protein were 0.369+/-0.035, 0.340+/-0.028, 0.097+/-0.020, respectively, in HMGB1-siRNAs transfection group, which were significantly lower than that in Lipofectamine 2000 alone group (0.553+/-0.051, P < 0.01). Of the 3 specific HMGB1-siRNAs, HMGB1-siRNA-3 (siRNAH3) had the highest inhibition rate (80%). The proliferation of HepG2 cells was markedly inhibited by siRNAH3 transfection. Compared to mock-transfection, siRNAH3 transfection dramatically suppressed the proliferation of HepG2 cells (P < 0.01). Moreover, siRNAH3 can induce apoptosis (P < 0.01).

CONCLUSIONsiRNA targeting HMGB1 mRNA can specifically reduce HMGB1 gene and protein expression. siRNAH3 can effectively suppress the proliferation and induce apoptosis of HepG2 cells.

Apoptosis ; Cell Proliferation ; HMGB1 Protein ; genetics ; Hep G2 Cells ; Humans ; RNA, Small Interfering