Objective To evaluate the effect of morphine on the growth of subcutaneous tumor of human gastric cancer cells in nude mice.Methods Thirty SPF male BALB/C nude mice,aged 4-5 weeks,weighing 15-20 g,in which the model of subcutaneous tumor of human gastric cancer cell line MGC-803 was established,were randomly divided into 3 groups (n=10 each) using a random number table:control group (group C),normal saline group (group N),and morphine group (group M).The mice in group C received no treatment.Morphine 20 mg/kg was intraperitoneally injected once a day for 14 consecutive days in group M,while normal saline 1.5 ml/kg was given instead of morphine in group N.The caliper was used to measure the tumor size every 2 days starting from 3 days after beginning of administration,and the relative tumor volume was calculated.The nude mice were sacrificed on 15th day,and the tumor tissues were obtained for determination of nuclear factor-kappa B activity and Bcl-2 and Bax protein and mRNA expression by real-time reverse transcriptase polymerase chain reaction and Western blot.Results Compared with group C,the relative tumor volume was significantly decreased,the activity of nuclear factor-kappa B in tumor tissues was significantly decreased,the expression of Bcl-2 protein and mRNA was significantly down-regulated,and the expression of Bax protein and mRNA was significantly up-regulated at each time point in group M (P＜0.05),and no significant change was found in each parameter mentioned above in group N (P＞0.05).Conclusion Morphine can inhibit the growth of subcutaneous tumor of human gastric cancer cells in nude mice,and the mechanism is associated with promotion of apoptosis in tumor cells.

Objective To study the anti,radiation effect of beta,carotene(?,C). Methods The different doses of ?,C (30,60 and 90 mg/kg bw) were orally administered separately to three groups of mice, and a control group was also arranged. The four groups of mice were exposed to 60Co,? rays after 30 days, then all groups were continuously orally administered, the survival rates, survival time, number of white blood cells(WBC), micronucleus frequency(MF), spermatocyte chromosome abberation, immune function, superoxide dismutase(SOD) activity were measured. Results The results showed that compared with control group, groups treated with?,C expressed lower range WBC decrease, stronger immune function, higher SOD activity, less MF, longer survival time and improved survival rates. Conclusion ?,C had radioprotective effect.

Translationally controlled tumor protein (TCTP)is a small protein highly conserved in a variety of eukaryotic species.TCTP is overexpressed in various tumor cells and has been implicated in the regu-lation of cellular processes including apoptosis,DNA repair and drug resistance.By reviewing the recent pro-gress of TCTP research,TCTP is becoming an important regulator of DNA repair and a new molecular target for tumor therapy.

AIM: To observe the changes of sarcoplasmic reticulum Ca~(2+)-ATPase (SERCA), phospholamban (PLB) during heart failure after acute myocardial infarction (AMI) in rats and the effect of carvedilol. METHODS: Rats were randomly assigned to normal control group, sham-operation group, AMI group and carvedilol (CAR) group. 6 weeks later, in vivo hemodynamic, morphometry and SERCA, PLB mRNA and protein expression of myocytes were measured in all animals. RESULTS: In comparison with sham-operation group, LV end diastolic pressure (LVEDP) and weight of ventricles were increased, while maximal rate of rise and fall (?dp/dt) of LV pressure were decreased in AMI group. After treatment with carvedilol, these parameters were all improved. The mRNA and protein expression of SERCA were downregulated (P0.05). CONCLUSIONS: The changes of SERCA and PLB may be the important mechanism of contractile dysfunction in heart failure after AMI. Carvedilol is effective in preventing LV dysfunction after AMI. The molecular mechanism may be related with normalization of SERCA expression.

Objective To investigate the effect of anti-syphilis treatment on the perinatal outcomes and neonatal prognosis in pregnant women complicated with syphilis.Methods One hundred and ninety eight pregnant women complicated with syphilis were collected from Chengdu Hospital of Infectious Diseases during January 2010 and January 2012,including 98 cases received standard treatment,59 cases received nonstandard treatment and 41 cases did not receive treatment.Pearson x2 and partition of chi-square were used for the comparison of pregnant outcomes,neonatal prognosis and negative rates of rapid plasma circle card test (RPR) among 3 groups.Results The incidence of adverse pregnancy outcomes,including miscarriage,prematurity,still birth and congenital malformation were 4.08％,27.12％ and 63.41％ in three groups,respectively.The incidence of congenital syphilis,low birth weight,asphyxia in infants and neonatal death raised in from standard-treatment group,nonstandard-treatment group to untreated group.Congenital syphilis rates were 2.04％,18.75％ and 35.29％ in three groups,respectively.RPR titers in newborns from mothers with high RPR titer (≥ 1 ∶ 8) in standard-treatment group were significantly lower than those in nonstandard-treatment group and untreated group (x2 =37.122,P ＜ 0.01).RPR negative rates were 100.00％,59.26％ and 25.00％ in three groups,respectively (x2 =18.839,P ＜ 0.01).Conclusion Standard anti-syphilis treatment can improve pregnant outcome,neonatal prognosis and reduce the incidence of congenital syphilis.

Objective To investigate the influence on right-to-left shunt (RLS) of patent foramen ovale with contrast transcranial Dopple (cTCD) in different positions.Methods 276 consecutive patients included ischemic cerebrovascular disease (ICVD),migraine,episodic vertigo and syncope.They underwent cTCD at rest and after Valsalva maneuver in 4 positions:upright sitting,supine,left lateral decubitus and right lateral decubitus,in random order.RLS was graded 0 (no microbubbles detected),1 (1-10microbubbles),2 (＞ 10 microbubbles but no curtain),and 3 (curtain,shower of microbubbles).Meanwhile,each patients was examined by contrast transthoracic echocardiography(cTTE),contrast transesophageal echocardiography (cTEE) or right heart catheterization.Results RLS detection rate was 48.2％ (133/276)through cTCD.They were also confirmed patent foramen ovale by 82 cases in cTTE,28 cases in cTEE,and 23 patients in right heart catheterization.At rest,RLS detection rate and microbubbles of 4 positions had no significant difference (P ＞0.05).After the Valsalva maneuver,RLS detection rate and microbubbles of each position were significantly higher than at rest (P ＜0.001),and the RLS detection rate and microbubbles in upright sitting were higher than that at rest(P ＜0.05).Conclusions RLS is best detected in the upright sitting position with Valsalva maneuver with cTCD.Upright sitting detection helps to improve the detection rate and microbubbles of RLS.

Objective To understand the self-care experiences of nurses during their life and work,in order to provide evidences for self-care improvement among nurses.Methods This qualitative study investigated 1 5 clinical nurses chosen by purposive sampling method.The participants were in-depth interviewed and the data were analyzed by Colaizzi method.Results The study distilled three major themes including motivations of the self-care,practice of self-care on the body,psychology and social network,and career and barriers of self-care. Conclusions Strengthening quality of nursing care provision can benefit from the improvement of nurses′self-care.Such improvement not only requires the motivation and practice of nurses themselves,but also requires multiple supports and helps from family,organization,and society.

Objective To establish transgenic mice model with over expression of neuritin in bone mar-row,for the further study on the function of neuritin protein in the treatment of peripheral neuropathy. Methods Two pairs of transgenic mice(loxp-stop-loxp-neuritin and lyz-Cre/+)were fed and propagated,the DNA from the mice tails extracted and the genotype of transgenic mice identified by PCR. The wild type mice with B6 were as-signed as the controls,and the immunofluorescence was used to detect the accuracy of the neuritinloxp/+ _lyz -Cre/+. Results The two trensgenetic homozygous mice had the ability to reproduce,and the hybrid offsprings were neuritinloxp/+_lyz-Cre/+,neuritinloxp/-_lyz-Cre/+,neuritinloxp/+_lyz-Cre/-,neuritinloxp/-_lyz-Cre/-. The re-sults were met with the Mendel′s law. The results of immunofluorescence showed that the expression of neuritin of neuritinloxp/+_lyz-Cre/+ mice in bone marrow was significantly higher than the wild type mice(P < 0.05). Con-clusion The PCR method is of high reliability for identification of sub pus genotype and the female neuritinloxp/+mice mating with the male lyz-Cre/+ ones is an effective way for obtaining the neuritinloxp/+_lyz-Cre/+ mice.

AIK is a novel cationic peptide with potential antitumor activity. In order to construct the AIK expression vector by Gateway technology, and establish an optimal expression and purification method for recombinant AIK, a set of primers containing AttB sites were designed and used to create the AttB-TEV-FLAG-AIR fusion gene by overlapping PCR. The resulting fusion gene was cloned into the donor vector pDONR223 by attB and attP mediated recombination (BP reaction), then, transferred into the destination vector pDESTl 5 by attL and attR mediated recombination (LR reaction). All the cloning was verified by both colony PCR and DNA sequencing. The BL21 F. coli transformed by the GST-AIR expression plasmid was used to express the GST-AIK fusion protein with IPTG induction and the induction conditions were optimized. GST-AIR fusion protein was purified by glutathione magnetic beads, followed by rTEV cleavage to remove GST tag and MTS assay to test the growth inhibition activity of the recombinant AIR on human leukemia HL-60 cells. We found that a high level of soluble expression of GST-AIK protein (more than 30% out of the total bacterial proteins) was achieved upon 0.1 mmol/L ITPG induction for 4 h at 37 °C in the transformed BL21 F. coli with starting OD₆₀₀ at 1.0. Through GST affinity purification and rTEV cleavage, the purity of the resulting recombinant AIK was greater than 95%. And the MTS assays on HL-60 cells confirmed that the recombinant AIK retains an antitumor activity at a level similar to the chemically synthesized AIK. Taken together, we have established a method for expression and purification of recombinant AIK with a potent activity against tumor cells, which will be beneficial for the large-scale production and application of recombinant AIK in the future.
Antimicrobial Cationic Peptides ; biosynthesis ; Antineoplastic Agents ; metabolism ; Escherichia coli ; metabolism ; Genetic Vectors ; HL-60 Cells ; Humans ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Sequence Analysis, DNA

Objective:To investigate the effects of ultrasound microbubble-mediated RasGAP SH3-binding protein 1 (G3BP1) transfection on the proliferation and migration of human liver cancer HepG2 cells.Methods:HepG2 cells were treated with ultrasound targeted microbubble destruction (UTMD) technology-mediated si-G3BP1. The expression of G3BP1 in HepG2 cell lines was detected by Western blot, and the silencing efficiency was analyzed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blot. HepG2 cell proliferation and migration were analyzed by flow cytometry, methyl thiazolyl tetrazolium (MTT) , EdU staining, colony formation experiment, wound healing experiment, Transwell experiment and Western blot.Results:After silencing G3BP1 in HepG2 cells, its mRNA and protein levels were significantly reduced (1.01±0.03 vs 0.27±0.03, 1.02±0.01 vs 0.33±0.04) ; UTMD-mediated si-G3BP1 could significantly reduce the proliferation rate (31.49±3.09 vs 12.51±1.02) , proliferation activity (1.20±0.13 vs 0.46±0.31) , EdU-positive cell rate (99.23±1.01 vs 36.75±4.03) , colony formation rate (96.45±1.21 vs 32.67±2.62) , scratch healing rate (97.58±1.04 vs 42.33±2.56) , migration rate (94.28±2.33 vs 39.36±2.51) and Ki67, Cyclin D1, Vimentin protein levels, increase E-cadherin protein levels.Conclusion:UTMD-mediated si-G3BP1 can inhibit the proliferation and migration of human liver cancer HepG2 cells.