Objective To identify the expression of a fusion gene GCA formed from GM-CSF gene and LMP2A gene of Epstein-Barr virus (EBV) in a recombinant BCG (rBCG) and to study its immunoge-nicity.Methods The rBCG was constructed to express the fusion gene GCA and the expressed products were detected by Western blot assay .ELISA was performed to measure specific antibody titers in serum sam-ples from mice immunized with rBCG .Lactate dehydrogenase assay was used to analyze the cellular immuni-ty of mice.A mouse model of EBV-positive gastric carcinoma was established to evaluate the therapeutic effects of rBCG.Results The target proteins of GM-CSF and LMP2A were successfully expressed in rBCG . The specific antibodies were detected in rBCG immunized mice as indicated by ELISA .The maximum anti-body titer reached 1 ∶27 900 [(326.5±7.8) pg/ml] as injection with rBCG 5×108/mouse.The rBCG in-duced cytotoxicity of cytotoxic lymphocytes (CTLs) to EBV-positive gastric carcinoma cells (GT39) (with a killing rate of 89.6%±6.8%) was significantly higher than that of control group (P<0.05) The sizes of tumor in PBS control group [(1964.0±548.7) mm3] and BCG group [(1268.65±72.4) mm3] were big-ger than those in rBCG group [(168.64±78.80) mm3].Conclusion The rBCG expressing GM-CSF and LMP2A fusion gene was successfully constructed .The rBCG could induce humoral and cellular immune re-sponses in mice and inhibit the growth of tumor .

494 ng/ml. The test showed a sensitivity of 100%, a negative predictive vale of 100%. Conclusion VIDAS D dimer is well sited as diagnostic tool for the exclusion pulmonary embolism.

Objective To explore the effect of NGAL knockdown by NGAL siRNA encapsulated with urocanic acid-modified chitosan nanoparticles (UAC).MethodsNGAL siRNA encapsulated by UAC and chitosan (CTS) respectively, which were then used to transfect human colon cancer cell lines HT29.The expression level of NGAL protein were detected by Enzyme Linked Immunosorbent Assay(ELISA).ResultsThe ELISA study revealed that the expression level of NGAL protein in UAC group(average 0.583μg/L) was significantly lower than in CTS group (average 0.772μg/L) and control group(average 1.071μg/L) (P<0.05).ConclusionThe NGAL expression of mRNA and protein in HT29 cells could be down-regulated by siRNA encapsulated by UAC.

Objective:To investigate the effects of total minimally invasive surgery (TMA) and mixed minimally invasive/open surgery (HMOA) on perioperative conditions and long-term efficacy of patients with rectal cancer.Methods:The clinical data of 240 patients with rectal cancer treated with minimally invasive surgery from January 2014 to August 2018 in Hangzhou Third People′s Hospital were retrospectively analyzed. Among them, 110 patients were treated with TMA (TMA group) and 130 patients were treated with HMOA (HMOA group). The relevant indexes of patients before and after surgery were collected and analyzed.Results:The operation time in TMA group was significantly longer than that in HMOA group: (312.5 ± 20.3) min vs. (210.8 ± 15.2) min, the length of hospital stay was significantly shorter than that in HMOA group: (4.0 ± 0.5) d vs. (6.8 ± 1.0) d, and there were statistical differences ( P<0.01); there were no statistical differences in low anterior resection and surgical procedures, ileostomy, open surgery, postoperative complications, reoperation, morphine dosage at 3 d after surgery and readmission between 2 groups ( P>0.05). Multivariate Cox analysis result showed that BMI ≥ 30 kg/m 2 ( OR=4.11, 95% CI 1.68 to 9.72, P<0.01), TMA ( OR=0.13, 95% CI 0.06 to 0.42, P<0.01), delayed bowel obstruction ( OR=13.6, 95% CI 1.59 to 110.56, P<0.05) and reoperation ( OR=15.32, 95% CI 5.52 to 42.56, P<0.01) were independent risk factors of prolonged hospital stay in patients with rectal cancer. The patients were followed up for 15 to 42 (29.5 ± 0.2) months, and there were no statistical differences in 3-year overall survival (OS) rate and 3-year disease-free survival (DFS) rate between HMOA group and TMA group (92.5% vs. 92.8% and 79.6% vs. 85.5%, HR=1.20 and 0.75, 95% CI 0.35 to 3.14 and 0.28 to 1.34, P=0.98 and 0.25). Conclusions:Patients with rectal cancer treated with TMA have the advantages of shorter hospital stay and shorter short-term effects compared with those treated with HMOA. However, the long-term effects of the two minimally invasive procedures are similar.

Objective:To evaluate the safety and efficacy of Delorme procedure for adults with full-thickness rectal prolapse.Methods:Clinical data of 17 adult patients suffering from full-thickness rectal prolapse undergoing Delorme procedure from June 2014 to May 2018 in Hangzhou Third Hospital were retrospectively analyzed. Patient characteristics, operative data, postoperative complications, recurrence of rectal prolapse, continence state and constipation state were evaluated.Results:Eleven patients were female, 6 patients were male with a mean age of (68 ± 9) years. Operations were successfully performed in these 17 cases. The operation time was (88 ± 16) minutes. The estimated blood loss during operation was (23 ± 9) ml. The postoperative time of hospital stay was (8 ± 1) d. Two complications in two patients were observed. There was no treatment related death. One recurrent case was observed during (16 ± 2) months follow-up. The preoperative and postoperative mean constipation score of five patients with fecal constipation were (23 ± 2) and (11 ± 3) respectively ( t = 9.51, P<0.01). The mean fecal incontinence score of six patients with fecal incontinence, before and after Delorme procedure, were (14 ± 2) and (6 ± 2) respectively ( t = 9.09, P<0.01). Conclusions:The Delorme procedure for adults with full-thickness rectal prolapse is a safe and effective surgery with less complications and low recurrence rate. The Delorme procedure may be one of the preferred option of perineal approach for adults with full-thickness rectal prolapse, but the long-term outcome of Delormer procedure and its effect on postoperative anal function need to be further studied.

Objective:To study the anti-tumor efficacy of endostar microbubble combined with focused ultrasound radiation in colon cancer liver metastases.Method:29 mice with colon cancer liver metastasis were randomly divided into four groups. Group 1(8 mice), as the control group. Group 2(7 mice) were treated only with ultrasonic radiation. Group 3 (7 mice) treated with the ultrasonic radiation combined with SonoVue microbubbles without carrying any medicine. Group 4(7 mice), treated with the ultrasonic radiation combined with microbubbles carrying endostar. The mice were sacrificed and the tumor specimens were weighted on the 12 days after ultrasound radiation. Immunohistochemistry was used to assess CD34 expression within the metastatic tumor.Results:The tumor weight in group 4 (0.79±0.49)g was significantly lower than that in group 1 (2.67±0.61)g, group 2 (2.60±0.60)g and group 3 (1.74±0.33)g ( F=20.629, P<0.01). The liver metastatic tumor weight in group 4(0.55±0.16) g was much lower than that in group 1 (1.47±0.22)g, group 2(1.42±0.28) g and group 3 (0.95±0.27)g ( F=23.758, P<0.01). There was no obvious difference among the four groups in the number of nodules of metastatic tumor in liver ( F=0.167, P=0.918). The level of CD34 in group 4 were (8 037±1 708) , significantly lower than that in any other group, ( F=15.779, P<0.01). Conclusion:Endostar microbubble combined with focused ultrasound radiation decreases tumor angiogenesis in liver metastasis, and inhibits the growth of both primary and metastatic tumor.

OBJECTIVETo explore the impact of neutrophil gelatinase-associated lipocalin (NGAL) knockdown by NGAL siRNA encapsulated with urocanic acid-modified chitosan nanoparticles (UAC) on the proliferation, migration and apoptosis of human colon cancer cells.

METHODSNGAL siRNA was encapsulated by UAC and chitosan (CTS) respectively, and then was transfected into human colon cancer cell lines HT29. The NGAL mRNA was detected by real-time quantitative PCR (RT-QPCR). Relationships of NGAL gene silencing with the proliferation, migration and apoptosis of HT29 cell were analyzed.

RESULTSUnder the fluorescence microscope, the transfection efficiency of siRNA in UAC group was (37.52±7.17)%, which was significantly higher than (11.32±3.39)% in CTS group (t=6.102, P=0.005). Forty-eight hours after transfection, RT-QPCR examination showed that the level of NGAL mRNA expression was 0.350 in UAC group and 0.529 in CTS group with significant difference (t=-3.743, P=0.02), meanwhile both levels were significantly lower as compared to control group(F=163.538, P<0.001). Proliferation analysis revealed that after silencing NGAL gene, proliferation rate of UAC group and CTS group was slightly lower than control group, and no significant differences were found (F=9.520, P=0.438). However, migration assay demonstrated that the 24-hour migration rate of UAC group and CTS group was significantly lower than that of control group (F=6.756, P=0.029), meanwhile the migration rate of UAC group was slightly lower than that of CTS group [(77.90±7.14)% vs. (87.67±3.98)%, t=-1.704, P=0.164]. Apoptosis detection revealed that the apoptosis rate in UAC group was significantly higher than that in CTS group and the control group 2 days after transfection [(15.800±1.054)% vs. (12.900±0.656)%, (11.933±1.914)%, F=7.004, P=0.027].

CONCLUSIONSThe encapsulated ability and transfection efficiency of chitosan modified by urocanic acid elevate significantly. Silencing NGAL gene by UAC carrier can down-regulate the expression of NGAL mRNA in HT29 colon cell line, inhibit their migration and facilitate their apoptosis.