In order to investigate the effect of antisense oligonucleotide (ASODN) of vascular endothelial growth factor C (VEGF-C) on lymphangiogenesis and angiogenesis of pancreatic cancer, antisense and scamble-sense oligonucleotide of VEGF-C were constructed, and the model of nude mice with orthotopically xenografted human pancreatic cancer cells (Panc-1) was established. Thirty nude mice were randomly divided into 3 groups: PBS control group (group A), scramble-sense control group (group B) and antisense group (group C). All nude mice were treated once every 2 days as 3 times per week, for 3 weeks (oligonucleotide 10 mg/kg every time). After treatments were completed, ELISA method was used to examine the concentration of VEGF-C in plasma and immunohistochemical method to examine microvessel density (MVD), lymphtic vessel density (LVD) of pancreatic cancer. The results showed that the expression of VEGF-C was inhibited significantly in group C. The concentrations were 237.5+/-41.5, 221.5+/-52.3 and 108.6+/-14.9 pg/mL in groups A, B and C respectively (P<0.01). LVD in groups A, B and C was 13.8+/-2.1, 12.4+/-1.9 and 4.2+/-1.6 respectively (P<0.01). MVD in groups A, B and C was 27.5+/-8.7, 25.9+/-4.2 and 19.4+/-5.6 respectively with no significant difference among the groups (P>0.05). It was suggested that VEGF-C ASODN decreased the expression levels of VEGF-C in nude mice with orthotopically xenografted human pancreatic cancer, and it could inhibit lymphangiogenesis, but had no significant effect on angiogenesis.

In order to investigate the correlation between protein expression of PTEN and the proliferation, infiltration, metastasis and prognosis in pancreatic cancer, immunohistochemical SP method was used to examine the protein expression of PTEN, PCNA, MVD, MMP-2, MMP-9 and TUNEL method to detect the levels of apoptosis of pancreatic cells in 41 pancreatic head cancers from regional pancreatectomy (RP) and 10 normal pancreatic tissues. The results showed that among 41 cases of pancreatic cancers, the positive staining of PTNE (39.02%) was significantly weaker than that in normal pancreatic tissues (P < 0.05). The levels of PCNA labeling index (LI), apoptotic index (AI), microvessel density (MVD), MMP-2 LI and MMP-9 LI were decreased gradually with the increase of the expression intensity of PTEN, and there was a significant difference in the above parameters among the patients having different expression levels of PTEN (P < 0.01 or P < 0.05). There was a negative correlation between the expression of PTEN and PCNA LI, MVD, MMP-2 LI, MMP-9 LI, and a positive correlation between AI and the expression of PTEN. The expression intensity of PTEN was correlated with the postoperative survival of the patients with pancreatic cancer (chi2 = 22.3400, P < 0.0001, RR = 2.030). It was suggested that the expression levels of PTEN protein were closely related with proliferation, infiltration and metastasis in human pancreatic cancer, and the expression of PTEN protein was one of the prognostic factors for pancreatic cancer following RP.

The effect of hypoxia inducible factor-1alpha (HIF-1alpha) on vascular endothelial growth factor C (VEGF-C) and the correlation between HIF-1alpha and lymphangiogenesis and lymph nodes metastases (LNM) in pancreatic cancer were investigated. Immunohistochemical SP method was used to detect the protein expression of HIF-1alpha and VEGF-C, and Lymphatic vessel density (LVD) was determined by stain of VEGFR-3, collagen type IV in 75 pancreatic head cancers from regional pancreatectomy (RP) during Dec. 2001 to Dec. 2003. The relationship between HIF-1alpha and VEGF-C, lymphangiogenesis, LNM was analyzed statistically. The results showed that the positive expressionrate of HIF-1alpha and VEGF-C in pancreatic cancer tissues was 48.00 % (36/75) and 65.33 % (49/75) respectively. In positive group of HIF-1alpha, the positive rate of VEGF-C and LVD, and LVD rate was 80.56 % (29/36), 13.22+/-3.76 and 88.89 % (32/36) respectively, and in negative group of HIF-1alpha, positive rate of VEGF-C and LVD was 51.28 % (20/39), 5.98+/-2.17 and 66.67 % (26/39) respectively (P<0.01 or P<0.05). It was suggested that HIF-1alpha could promote the expression of VEGF-C, lymphangiogenesis and LNM in pancreatic cancer.

Objective:To evaluate the clinical effect of retroperitoneoscopic renal pedicle lymphatic disconnection via extra-adipose capsule in the management of chyluria and to discuss the management of its complications.Methods:From August 2013 to June 2008,five patients with chyluria were admitted. All the patients were female,aged from 26 to 73 years,and disease course from 1 to 10 years.All the five patients had complained of intermittently voiding milky urine with varying degrees of weight loss,and fa-tigue,of whom two presented with flank pain and one with anemia.Their urine chyle tests were con-firmed to be positive.Preoperative cystoscopy found that chyluria was from the left side in 3 cases,and from the right side in 2 cases.Their proteinuria ranged from +to ++++.All the cases had been treated with the modified procedure by which lymphatic ligation was performed to hilar vessels and proximalureter via extra-adiposecapsule without disconnection of perirenal fat tissues.The operation time,intraoperative blood loss,postoperative intestinal function recovery,catheter time,drainage tube removal time and com-plications during operation were collected.Results:All the five cases were performed successfully.The operation time ranged from 75 to 170 minutes,mean (126.0 ±39.6)minutes,with the intraoperative blood loss 20 to 60 mL,mean (38.0 ±16.4)mL,and the postoperative intestinal function recovery time 1 to 3 days,mean (1.9 ±0.4)days.The catheter time was 1 to 4 days,mean (2.1 ±0.3)days and the drainage tube removal time ranged from 3 to 15 days,mean (9.3 ±1.8)days.Postoperatively lym-phorrhagia was found in two cases.No renal vessels injury occurred during operation.Chyluria of all the patients disappeared on the operation day with negative chyluria test after surgery.Furthermore,urine test revealed that proteinuria was totally negative.No recurrence was detected in our patients in the 9 to 31 months’follow-up.Conclusion:This modified procedure does not have to disconnect perirenal fat tissues and nephropexy during operation.It was characterized with shorter operation time,definitive effect and fewer complications.

Objective To investigate the role of erythropoietin(EPO)in relieving the injury of human renal tubular cells (HK-2) induced by postasphyxial-serum of neonates.Methods Human renal proximal tubular cell(HK-2) was used as the target cell.The experiment was designed as control group, asphyxia group,and group of pretreatment with EPO. The attacking concentration of serum was 200 mL/L,then the changes of morphology were observed under inverted microscope,and the cell viability was measured by 3-(4,5-dimethy lthiazcl-2-yl)-2,5-diphenyl-tetazolium bromide(MTT) methods,and the leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.Results Compared with control group,the change in morphology of HK-2 was most serious and obvious,and the leakage rate of LDH increased significantly,and the cell viability decreased obviously in asphyxia group.But compared with asphyxia group,the change in morphology of HK-2 was obviously improved,and the leakage rate of LDH decreased and the cell viability increased in group of pretreatment with EPO in a dose-dependent manner except the group of 1 IU/mL.Conclusion EPO can play the role in relieving the injury of renal tubular cells induced by postasphyxial-serum in neonates.

Humans ; Enhanced Recovery After Surgery ; Stomach Neoplasms/surgery* ; Length of Stay ; Laparoscopy ; Postoperative Complications ; Gastrectomy

This retrospective analysis compared standard regimen of doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) with the dose-dense ABVD regimen (ABVD-21) in terms of efficacy and toxicity. Patients who had early-stage unfavorable or advanced Hodgkin's lymphoma (HL) according to German Hodgkin Study Group criteria from March 1999 to February 2011 were analyzed for treatment response, long-term survival and hematological toxicity. There were 85 patients in the ABVD-21 group and 118 patients in the ABVD group respectively. The complete remission rates after completion of treatment were 92.9% and 90.7% for ABVD-21 and ABVD, respectively. During a median follow-up period of 62 months, no significant difference was found in projected 10-year progression-free survival (PFS) and overall survival (OS) rates (84.7% and 94.1% respectively for ABVD-21; 81.4% and 91.5% for ABVD). Subgroup analyses showed that ABVD-21 was significantly better than ABVD for patients with IPS≥3 in terms of PFS and OS rates. Grade 3 to 4 leukopenia (51.8% vs. 28.8%, P=0.001) and neutropenia (57.6% vs. 39.0%, P=0.009) were more common with ABVD-21. We were led to conclude that dose-dense ABVD did not result in better tumor control and overall survival than did ABVD for early-stage unfavorable HL. However, patients at high risk, for example, with IPS≥3, may benefit from dose-dense ABVD.

In order to investigate the effect of antisense oligonucleotide (ASODN) of vascular endothelial growth factor C (VEGF-C) on lymphangiogenesis and angiogenesis of pancreatic cancer, antisense and scamble-sense oligonucleotide of VEGF-C were constructed, and the model of nude mice with orthotopically xenografted human pancreatic cancer cells (Panc-1) was established. Thirty nude mice were randomly divided into 3 groups: PBS control group (group A), scramble-sense control group (group B) and antisense group (group C). All nude mice were treated once every 2 days as 3 times per week, for 3 weeks (oligonucleotide 10 mg/kg every time). After treatments were completed, ELISA method was used to examine the concentration of VEGF-C in plasma and immunohistochemical method to examine microvessel density (MVD), lymphtic vessel density (LVD) of pancreatic cancer. The results showed that the expression of VEGF-C was inhibited significantly in group C. The concentrations were 237.5±41.5, 221.5±52.3 and 108.6±14.9 pg/mL in groups A, B and C respectively (P＜0.01). LVD in groups A, B and C was 13.8±2.1, 12.4±1.9 and 4.2±1.6 respectively (P＜0.01). MVD in groups A, B and C was 27.5±8.7, 25.9±4.2 and 19.4±5.6 respectively with no significant difference among the groups (P＞0.05). It was suggested that VEGF-C ASODN decreased the expression levels of VEGF-C in nude mice with orthotopically xenografted human pancreatic cancer, and it could inhibit lymphangiogenesis, but had no significant effect on angiogenesis.

OBJECTIVETo investigate the effects of small interfering RNA (siRNA) recombinant expression vector targeting survivin gene on chemotherapy sensitivity of human colon cancer cells to 5-fluorouracil.

METHODSsiRNA recombinant expression vector targeting survivin gene was constructed and transfected into human colon cancer cell lines LOVO. After 48 hours of transfection, cells were harvested for analysis of survivin mRNA and protein expressions using RT-PCR and Western blot. In addition, after human colon cancer cell lines were treated with Survivin siRNA and/or 5-fluorouracil, MTT assay and flow cytometry were used to analyze cell proliferation and apoptosis.

RESULTSRestriction endonuclease analysis confirmed that siRNA recombinant expression vector targeting survivin gene was successfully constructed. Inhibitory ratios of survivin mRNA and protein expressions by Survivin siRNA were 36.33% and 44.65%, respectively. Survivin siRNA combined with 5-fluorouracil significantly increased the cell proliferation inhibitory ratio and apoptosis ratio compared with 5-fluorouracil treating alone (P<0.05).

CONCLUSIONThe siRNA recombinant expression vector targeting survivin gene can inhibit the expression of survivin gene, and enhance chemotherapy sensitivity of human colon cancer cells to 5-fluorouracil.

Antimetabolites, Antineoplastic ; therapeutic use ; Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; drug therapy ; genetics ; metabolism ; Fluorouracil ; therapeutic use ; Genetic Vectors ; genetics ; metabolism ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; metabolism

In order to maximize the efficiency and versatility of the vector-based siRNA approach, we have developed a novel siRNA expression vector containing multiple tandem siRNA cassettes to investigate the synergistic inhibitory effect of it on human colon cancer cells proliferation. Multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes was constructed and transfected into human colon cancer cell. The effect of multiple siRNA recombinant expression vector was detected by RT-PCR, Western blotting and flow cytometry. It was confirmed by restriction endonuclease and sequence analysis that multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes was constructed successfully. Livin and Survivin genes inhibition ratio of Livin and Survivin siRNA at mRNA levels were 27.90% and 32.24%, at protein levels were 22.28% and 40.86%, the apoptotic ratio was (11.69 +/- 1.37) %, but the synergistic effect was weaker than Livin and Survivin RNA interference, respectively. The multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes has been constructed successfully. It can inhibit the expression of Livin and Survivin genes in human colon cancer cells, but the synergistic effect was weaker than Livin and Survivin RNA interferences, respectively.
Adaptor Proteins, Signal Transducing ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; Neoplasm Proteins ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection