Objective To investigate the effects of selective cyclooxygenase-2(COX-2) inhibitor nimesulide on the proliferation of colon adenocarcinoma cells in vitro and the expression of matrix metalloproteinase-2(MMP-2).Methods The human colon cancer cell lines HT-29 and HCT-116 were employed in the study,grouped as nimesulide group,DMSO control group and blank control group.After treatment with nimesulide,the inhibitory effect of nimesulide on the proliferation of cancer cells was quantified by MTT assay,and the expression of MMP-2 in the cells was detected by quantitative zymography.Results Nimesulide inhibited the proliferation of HT-29 and HCT-116 cells in time and dose-dependent manners.The inhibitory effect on HT-29 cells was stronger than that on HCT-116 cells.Nimesulide down-regulated the MMP-2 expression in HT-29 cells,whereas the expression in HCT-116 cells remained unchanged.Conclusion Nimesulide can obviously inhibit the growth of colon cancer HT-29 cells with positive COX-2 protein,suggesting that nimesulide may down-regulate the expression of MMP-2 by inhibiting the activity of COX-2.

Objective To evaluate the potential roles of celecoxib on proliferation and cell cycle progression of colon adenocarcinoma cells and on the hepatic metastasis of nude mice. Methods The human colon cancer cells HT-29 and HCT-116 were employed in the study. After treatment with celecoxib, the inhibitory effects of celecoxib on the proliferation of cancer cells were quantified by MTT assay, and the cell cycle progression was detected by flow cytometry, tumor cells were inoculated in nude mice, and the hepatic metastasis was detected. Results ①Celecoxib inhibited the proliferation of the tumor cells in time and dose-dependent manners (P

According to the limited class hours and requirements of medicinary major curriculum application,this paper attempts to make some choice of physical chemistry teaching contents and emphasize on the first class,and also to explore bilingual teaching of partial chapters and experimental teaching.

SUMMARY Herewereportedapatientwithwarfarin-inducedautoimmunehepatitis(AIH),andex-plored new concerns for the pharmaceutical care of warfarin.A 57-year-old woman was admitted to hospi-tal for repeated anorexia,abdominal pain and abnormal liver function.She received prosthetic heart valve replacement because of rheumatic heart disease,and had started warfarin medication since 2 years before.Her liver function was elevated with highest alanine aminotransferase 861 U/L, aspertate aminotransferase 604 U/L,and total bilirubin 1 06.7 μmol/L.Her anticoagulant therapy was switched to low molecular weight heparin and the liver function returned to normal.The liver function was elevated when she started to take warfarin again.The patient was then on liver protection therapy,and warfarin was stopped again for the liver biopsy for diagnosis reason.Through medication consultation and evalua-tion,pharmacists were invited to work together with the physicians and helped to differentiate the reason for abnormal liver function, and provided therapeutic suggestions. Also the pharmacists gained experiences in the treatment of AIH,and discovered a new and severe adverse drug reaction for warfarin. In treating this case,the pharmacists’active involvement into the treatment and evaluation of the effect on the patient reflected the advantage and importance of the multidisciplinary cooperation for pharmacists and physicians when complex diseases are faced.

Objective To compare detection of STAT3 and CEA by nest RT-PCR and peritoneal lavage cytology examination(PLC) in predicting peritoneal micrometastasis in gastric carcinoma patients. Methods Peritoneal washings of 66 patients with gastric carcinoma and 12 patients with benign gastric diseases were collected to detect the mRNA level of STAT3 and CEA by nest RT-PCR.PLC were carried out by H-E staining.Results The positive rates of STAT3 mRNA and CEA mRNA in peritoneal washings were 43.9% and 48.5%,respectively.The positive rates of STAT3 mRNA and CEA mRNA were significantly higher than those of PLC(31.8%).The positive rates of STAT3 mRNA and CEA mRNA were concurrently increased with the depth of tumor invasion,TNM staging and pathogenesis.Conclusion Detecting STAT3 mRNA in peritoneal washings by nest RT-PCR can improve the sensitivity of free tumor cell detection in peritoneal washings,and may be used to predict peritoneal micrometastasis of gastric carcinoma.

Background Pulmonary fibrosis currently lacks screening and diagnostic methods in the early stages and effective treatments in the later stages, so there is an urgent need to explore the mechanisms and develop targeted treatments. Objective To screen the expression of differentially expressed circular RNA (circRNA) hsa_circUCK2 under pathological conditions and to explore its effect on pulmonary fibrosis. Methods In the cell-based experiments, hsa_circUCK2 was knocked down in HPF-a cells using small interfering RNA (siRNA), and HPF-a cells were stimulated by TGF-β1. Four groups were set up: si-NC group, si-circUCK2 group, si-NC+TGF-β1-treated group, and si-circUCK2+TGF-β1-treated group. Western blot assay was used to detect the expression of fibronectin (FN1) in HPF-a cells of the four groups, scratch assay was used to detect the migration ability of HPF-a cells, and CCK-8 assay was used to detect the proliferation ability of HPF-a cells in the two groups with TGF-β1 stimulation, the si-NC+TGF-β1-treated group and the si-circUCK2+TGF-β1-treated group. In the animal experiments, forty-eighty healthy SPF-grade male C57BL/6 mice were randomly divided into four groups: saline+si-con group, saline+si-circ_0000115 group, SiO₂+si-con group, and SiO₂+si-circ_0000115 group. Mouse lung circRNA mmu_circ_0000115 (mouse homolog of hsa_circUCK2) was knocked down by tracheal drip injection of siRNA, and a mouse lung fibrosis model was constructed by tracheal drip injection of SiO₂ suspension (0.2 g·kg⁻¹, 50 mg·mL⁻¹) after 48 h. Real-time fluorescence quantitative PCR was used to detect the knockout efficiency in each organ of the mouse, Western blot was applied to detect the expression of type I collagen α2 (COL1A2) in the lung tissues, and Sirius red was used to detect collagen synthesis in the lung tissues. Results In the cell-based experiments, after the knockdown of hsa_circUCK2, the Western blot results showed that the expression level of the FN1 protein in TGF-β1-stimulated HPF-a cells was significantly down-regulated (P <0.05); the CCK-8 assay and cell scratch assay showed that the down-regulation of hsa_circUCK2 gene significantly inhibited the proliferation and migration of HPF-a cells (P<0.01). In the animal experiments, the real-time fluorescence quantitative PCR results showed that among the detected organs, mmu_circ_0000115 was significantly knocked down only in the lung tissues (P<0.0001); the Western blot results showed that knocking down mmu_circ_0000115 significantly reduced the COL1A2 protein expression level when compared with the SiO₂+si-con group (P<0.0001); the Sirius red results showed that knocking down mmu_circ_0000115 significantly reduced collagen production and deposition in lung tissues of mice in the model group. Conclusion Knockdown of hsa_circUCK2 inhibits fibroblast activation and reduces collagen deposition in lung fibrosis model mice. It is suggested that the hsa_circUCK2 is involved in the process of pulmonary fibrosis and may be a potential therapeutic target for pulmonary fibrosis.

ObjectiveTo investigate the effect of propofol on cyclooxygenase-2 (COX-2) expression in hippocampal neurons in depressed rats after electroconvulsive therapy (ECT).MethodsFifty 2-3 months old male SD rats weighing 200-250 g were randomly divided into 5 groups (n =10 each):group control (group C) ; group depression (group D); group propofol (group P); group ECT (group E) and group propofol + ECT (group PE).Depression was induced by separation and chronic unpredictable mild stres in groups D,P,E and PE.Groups P and PE received intraperitoneal pro pofol 80 mg/kg.Groups E and PE received ECT at 5 min after IP normal saline 8 ml/kg and propofol 80 mg/kg respectively once a day for 7 consecutive days.The learning and memory function was assessed by using Morris water maze test before (baseline) and after depression was induced and ECT.The animals were then sacrificed and their brains removed for detection of COX-2 mRNA expression in hippocampus (by RT-PCR).Results In group D depression significantly prolonged evasive latency and decreased swimming time percentage in platform quadrant and up-regulated COX-2 mRNA expression as compared with group C.In group E ECT further prolonged evasive latency and up-regulated COX-2 mRNA expression in depressed rats.In group PE propofol pretreatment attenuated ECT-induced impairment of learning-memory function and increase in COX-2 mRNA expression as compared with group E.ConclusionPropofol can ameliorate the decrease in learning and memory function induced by ECT in depressed rats by inhibiting COX-2 expression in hippocampal neurons.

ObjectiveTo observe the subcutaneous and intraperitoneal insulin injection's effect of the level of oxygen free radicals of type 2 diabetes model.MethodsC57BL/6J mice were chosen as normal control group (C group,n=9).KK mice were randomly divided into intraperitoneal injection of insulin group (i.p.group,n=9),the subcutaneous insulin group (s.c.group,n=9) and untreated group (U group,n =9).The i.p.group and the s.c.groups were given certain amount of insulin (insulin injecta and protamine insulin injecta by volume ratio of 2:1 mixture)for one month,maintained the GLU at normal levels (6±1.5) mmol/L.SOD,GSH-PX activity and MDA content of serum,liver,kidney and heart in each group were detected.Results The liver,kidney,heart and serum's SOD and GSH-PX activity significantly reduced and MDA content significantly increased in the U group.Both kinds of delivery methods could increase serum SOD and GSH-PX activity and reduce the content of MDA to the normal control group level,but the intraperitoneal injection had stronger effect.Two kinds of delivery methods could both reduce the MDA content of liver,and had almost the same effect; but the subcutaneous injection group had better effect on increasing the liver's SOD activity,and the intraperitoneal injection had better effect on increasing liver's GSH-PX activity.Intraperitoneal injection had better effect on reducing kidney' s MDA content and increased SOD activity.Two kinds of delivery methods had the same effect on reducing the heart's MDA content.Conclusion The two delivery methods can both make the MDA levels of KK mice in serum,heart,liver and kidney fall to as normal as that of control group,but the two delivery methods have different ways of improving the antioxidant capacity in different organs.Intraperitoneal injection can reduce MDA content in serum and kidney better.

Objective To investigate the effect of propofol anesthesia on electroconvulsive therapy (ECT)-induced hyperphosphorylation of Tau protein in hippocampus in depressed rats.Methods Thirty-two female WYK rats in which the total score was 30-120 after Open-field test,aged 24 weeks,weighing 200-250 g,were randomly divided into 4 groups ( n =8 each):control group (group C),propofol group (group P),ECT group (group E)and propofol + ECT group (group PE).In groups C and E,the animals received intraperitoneal normal saline 5 ml,and in addition the animals received ECT 15 min later in group E.In groups P and PE,the animals received intraperitoneal 100 mg/kg propofol 5 ml,and in addition the animals received ECT 15 min later in group PE.The learning and memory function was assessed by Morris water maze test at 24 h after ECT.The animals were sacririced at 6 h after Morris water maze test and the hippocampal tissues were removed for determination of the expression of phosphorylated Tau protein.Results Compared with group C,the escape latency was significantly prolonged,the swimming time was significantly shortened in groups P,E and PE,the expression of phosphorylated Tau protein in hippocampus was down-regulated in group P,and the expression of phosphorylated Tau protein in hippocampus was up-regulated in group E ( P ＜ 0.05).Compared with group E,the escape latency was significantly shortened,the swimming time was significantly prolonged,and the expression of phosphorylated Tau protein in hippocampus was down-regulated in group PE (P ＜0.05).Conclusion The mechanism by which propofol anesthesia improves cognitive impairment induced by ECT may be related to inhibition of hyperphosphorylation of Tau protein in hippocampus in depressed rats.

OBJECTIVETo investigate the inhibitory effect of puerarin on invasive and metastatic abilities of tumor cells, and its possible mechanism through observing its impacts on the migratory, adhesive and invasive capacities of human oophoroma cells HO-8910 to the artificial recombined basement membrane.

METHODSExpression of estrogen receptor (ER) in HO-8910 cells was detected using PCR assay. Effects of puerarin on HO-8910 proliferation was detected with MTT assay; on its adhesion potential was tested with cell-matrigel adhesion assay, and on invasive and migratory capacities were measured with Transwell matrigel invasion assay and Transwell motility assay respectively.

RESULTSER was positively expressed in HO-8910 cells. After being treated with 20 micromol/L puerarin for 12 h, the adhesive test showed that OD value in the tested group was significantly lower than that in the control (P < 0.01), the inhibiting rate reached 50.63%; and the Transwell assay showed a significant lowering of penetrated cells (P < 0.01), the inhibition rate for invasion was 38.59% and that for motility migration 40.63%. The number of penetrated cells was lower in the group intervened with combination of Puerarin and estrogen than in the group intervened with estrogen alone, 33.40 +/- 3.30 vs 48.05 +/- 3. 56 for invasion and 35.35 +/- 3.03 vs 52.45 +/- 1.04 for motility (all P < 0.01).

CONCLUSIONPuerarin can inhibit the adhesion, invasion and migration of HO-8910 cells, plays an antagonist effect against the stimulation of estrogen on the malignant behavior of tumor cells.

Cell Adhesion ; drug effects ; Cell Line, Tumor ; Cell Movement ; drug effects ; Female ; Humans ; Isoflavones ; pharmacology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Ovarian Neoplasms ; pathology ; Receptors, Estrogen ; metabolism