Objective We have examined tumor suppressor gene p53 from N- methyl - N- nitrosourea (MNU ) induced the col - orectal tumors in rats. Methods PCR, SSCP and DNA sequence analysis. Results In colorectal tumors induced by MNU, the mobil- ity shifts of p53 gene mutation were observed, p53 gene mutation was a deletion at third position C of codon 244 in exon7. Conclu- sion The present study suggested that MNU induces rat p53 mutations in the colorectal tumors.

Objective To evaluate the efficacy and safety of bilateral transversus abdominis plane block (TAPB)combined with bilateral rectus sheath block (RSB)in abdominal surgery. Methods Ninety ASA Ⅰ or Ⅱ patients,35 males,55 females,aged 19-79 years,with body mass index 18-30 kg/m2 ,scheduled for elective laparoscopic cholecystectomy were randomly divided into three groups(n=30):ultrasound-guided bilateral TAPB combined with bilateral RSB group (group TR),ultrasound-guided bilateral TAPB group (group T),patient-controlled intravenous analgesia (PCIA)group (group P).In group TR,ultrasound-guided bilateral TAPB were performed with 20 ml of 0.22% ropivacaine mesylate injection in each side and ultrasound-guided bilateral RSB were per-formed with 10 ml of 0.22% ropivacaine mesylate injection in each side before surgery.In group T, ultrasound-guided bilateral TAPB were performed with 20 ml of 0.22% ropivacaine mesylate injection in each side and ultrasound-guided bilateral RSB were performed with 10 ml of NS in each side before surgery.In group P,ultrasound-guided bilateral TAPB were performed with 20 ml of NS in each side and ultrasound-guided bilateral RSB were performed with 10 ml of NS in each side before surgery, and PCIA was applied in group P.BP,HR,SpO2 were observed when patients were sent into the op-erating room, 2 minutes before trocar puncture, and 2 minutes after trocar puncture, the consumption of propofol and remifentanil used during the surgery were recorded.The score of visual analogue scale (VAS)during rest and movement were recorded at 2,6,12,24 h after surgery.The patient analgesia satisfaction and the adverse reactions were recorded.Results Compared with group T and group P,group TR had less change of BP before and after trocar puncture(P <0.05).The VAS score was significantly lower in group TR after operation(P <0.05).There were no statistical significant differences of VAS score at 24 h after operation among the three groups.The patient anal-gesia satisfaction was significantly better in group TR than other two groups (P < 0.05 ). Conclusion Ultrasound-guided bilateral transversus abdominis plane block combined with bilateral rectus sheath block is of safety and much efficacy of postoperative analgesia in patients undergoing laparoscopic cholecystectomy.

BACKGROUNDOver-expression of RAB5A gene has been proved to be associated with neoplasia metastasis. This study is to explore the effect of RAB5A gene on invasion and metastasis of human lung adenocarcinoma cell lines.

METHODSConstituted basement membrane invasion technique, adhesion capability of tumor cell assay, the chemotactic migration of tumor cells assay, and gelatinases SDS-PAGE analysis method were used to detect the changes of invasive and metastatic capability of Anip973 (with high metastatic capability) and its parent AGZY83-a cell lines (with low metastatic capability).

RESULTSAfter AGZY83-a cells were transfected by PcDNA3.1-RAB5A plasmid, its invasion was significantly increased (t=24.36, P < 0.000 5); adhesion capability of cell was promoted (P < 0.05); the chemotactic migration of cells was higher than that of the parent lines (t=14.18, P < 0.000 5); and the activity of gelatinases secreted from transfected AGZY83-a was enhanced. The invasion of the transfected Anip973 cells with PcDNA3-AntiRAB5A was lower (t= 16.510 4, P < 0.002 5); adhesion capability of cell was decreased (P < 0.05); the chemotactic migration of cells was lower than that of the parent lines (t=6.062, P < 0.005); and the activity of gelatinases was obviously decreased.

CONCLUSIONSThis study in vitro indicates that over-expression of RAB5A genes plays an important role in tumor invasion and metastatic phenotype formation of human lung adenocarcinoma cells; and antisense RNA can interrupt the translation of RAB5A gene.

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 characteristics of neuropsychological factors in patients with persistent postural-perceptual dizziness(PPPD) and provide the basis for the psychosomatic comprehensive treatment. Methods: Cartel Personality Test (16PF), Symptom Checklist 90 (SCL-90), HAMA, HDMD, SAS and SDS were used to evaluate personality and mental state in patients with PPPD(PPPD group, n=65) and control group(n=63). Dizziness handicap inventory(DHI) was used to evaluate the degree of vertigo.The correlation analysis was carried out between the DHI scores and 16-PF, SCL-90 factor scores. Results: (1)16PF factor scores: the factor scores of assertiveness(8.50±1.84), excitability (6.59±1.73), boldness (7.46±1.78), sensitivity (7.25±1.79), doubtfulness (6.55±1.74), fantasy(6.20±1.60), anxiety(7.67±1.61) and tension(6.81±1.67)in PPPD group were higher than those in the control group, and the differences were statistically significant (all P<0.05). The gregariousness (4.38±1.65), intelligence (4.51±1.67), stability (3.51±1.75), independence (4.39±1.56) and self-discipline (4.70±1.82) factor scores in PPPD group were lower than those in the control group, and the differences were statistically significant (all P<0.05). (2)SCL-90 factor scores: the factor scores of somatization(1.62±0.40), anxiety (1.64±0.56), interpersonal sensitivity (1.79±0.42), terrifying(1.71±0.53), total points(150.77±21.60), total average score (1.62±0.51) in PPPD group were higher than those in control group (all P<0.05). There were no differences in obsessive-compulsive (1.50±0.55), depression (1.45±0.44), hostility (1.69±0.60), paranoia (1.76±0.53), somatization (1.42±0.49) and psychotic (1.29±0.35) between PPPD group and the control group (all P>0.05). (3)The factor scores of HAMA(9.08±1.77) and SAS(37.88±2.96)in patients with PPPD were higher than that in the control group, and the differences were statistically significant (all P<0.05). There was no significant difference in HAMD (6.19±2.82) and SDS (36.36±4.71) scores between PPPD group and control group (all P>0.05). (4)The DHI scores were positively correlated with assertiveness, sensitivity, tension and doubtfulness factors of 16PF.The DHI scores were positively correlated with somatization, interpersonal sensitivity, anxiety and terrifying factors of SCL-90. Conclusion Patients with persistent postural-perceptual dizziness suffer from personality changes, mental disorders and anxiety disorder.

Objective To evaluate the efficacy of ultrasound-guided transversus abdominal plane (TAP) and posterior rectus sheath (PRS) block for postoperative analgesia in the patients undergoing radical resection for gastric cancer.Methods One hundred twenty patients of both sexes,aged 18-64 yr,with body mass index of 19-25 kg/m2,of American Society of Anesthesiologists physical status Ⅱ or Ⅲ,scheduled for elective radical resection for gastric cancer,were divided into 2 groups (n =60 each) using a random number table:control group (group C) and ultrasound-guided TAP and PRS block group (group T+R).Bilateral TAP (0.375％ ropivacaine 0.5 ml/kg was injected) and PRS block (0.375％ ropivacaine 0.3 ml/kg was injected) were performed before induction of general anesthesia in group T+R.Patient-controlled intravenous analgesia was provided to all the patients after surgery in the two groups,and the visual analog scale score at rest and during activity was maintained less than 4 within 48 h after surgery.The requirement for rescue analgesia within 48 h after surgery and occurrence of adverse reactions during the analgesia period were recorded.Results Compared with group C,the requirement for rescue analgesia within 48 h after surgery and incidence of nausea and vomiting were significantly decreased in group T+R (P＜0.05).Conclusion Ultrasound-guided TAP and PRS block provides better efficacy for postoperative analgesia with less adverse reactions in the patients undergoing radical resection for gastric cancer.