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Objective To detect circulating tumor cells (CTCs) in the peripheral blood of patients with pancreatic cancer using a new nano microfluidic chip and to explore the relationship between CTCs and clinicopathological feature,postoperative recurrence and prognosis of pancreatic cancer.Methods The peripheral blood samples of 53 patients with pancreatic cancer who underwent curative resection in the second affiliated hospital of Jiaxing college of medicine were collected from January 2015 to January 2017.The CTCs from peripheral blood were detected by novel nano microfluidic chip.The cut-off value for CTCs-positive and negative groups was 1 CTC.The relationship between CTCs positive and postoperative recurrence and prognosis of pancreatic cancer were evaluated.Results The number of CTCs for 23 pancreatic cancer of 53 patients ranged from 5 to 196 per ml,and the mean number was 78.5 ± 44.7 per ml;the rate of CTCs-positive patients was 43.4％ (23/53).There were significant correlation between CTCs with vascular invasion (P =0.001),but but CTCs was not correlated with the gender,age,the presence of clinical symptoms,tumor size,pathological type,lymph metastasis and TNM stage.31 patients had tumor recurrence,and the rate of tumor recurrence was 58.5％.Among them,there were 13 cases with tumor local recurrence,10 cases with tumor distant metastasis (including liver,lung,kidney,etc.) and 8 cases with both tumor local recurrence and distant metastasis.The median recurrence free survival time of all patients was 14.0 months (13.0-17.0) and the median overall survival time was 19.0 months (15.5-24.0).The cumulative one-year and two-year recurrence free survival rate were 66.9％,12.2％ for patients with CTCs-positive and 88.3％,42.2％ for CTCs-negative patients,and the differences were statistically significant (both P ＜ 0.05).The cumulative one-year and two-year overall survival rate were 85.4％,33.6％ for patients with CTCs-positive and 96.3％,62.2％ for CTCs-negative.There was no difference in statistics in cumulative one-year overall survival rate and with a statistically significant difference in cumulative two-year overall survival rate (P =0.028).Conclusions Peripheral blood CTCs of pancreatic cancer can be effectively detected by a novel nano microfluidic chip.There were significant correlation between CTCs with vascular invasion and survival time after surgery.CTCs may be a potential prognostic indicator of the postoperative recurrence and prognosis of pancreatic cancer.
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Objective To investigate the effects of chemokine like factor 1 (CKLF1) gene on the proliferative activities and osteogenic potentials of hip ligaments of ankylosing spondylitis (AS) in situ and in vitro. Methods Normal and AS hip ligament specimens were collected from 6 patients with femoral neck fracture and 4 AS patients with severe hip deformities. Ligament specimens were exposed to type Ⅱ colla-genase and obtained a single cell suspension, while phase contrast microscopy and anti-vimentin immuno- fluorescence staining (IFC) were applied to observe the cells. The specimens and fibroblasts were divided and cultured in situ and in vitro respectively, and the recombinant adeno-associated virus (rAAV)-lacZ (E. coli beta-galactosidase gene)and rAAV-hCKLF1 (human CKLF1 cDNA cloned in rAAV-lacZ in place of lacZ) were transduced for 21 days. Cell proliferation (cellularity), secretion of pro-inflammatory cytokines, expression of CKLF1 and CCR4 genes were detected by the water-soluble tetrazolium (WST-1) assay and Hoechst 33258 test (DNA content), enzyme-linked immunosorbent assay (ELISA), IFC test and fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCR), respectively. Statistical analysis significance was conducted using the Student's t test and one-way analysis of variance (ANOVA) (LSD) test where appropriate. Results The second passage of normal and AS cells were positive for anti-vimentin, indicating that the cells were fibroblasts. After transducing with rAAV-hCKLF1 for 21 days, cellularity, WST-1 and Hoechst 33258 assays illustrated that CKLF1 gene transfer promoted cell proliferation (compared with the non-viral transduction and lacZ groups, F=6.98, 64.32, 115.91, P<0.05 or P<0.01). Overexpression of CKLF1 gene enhanced the secretion of pro-inflammatory cytokines (interleukin-6 and tumor necrosis factor-alpha) and the expression of bone-specific extracellular matrix proteins (osteopontin and osteocalcin) (F=34.57, 8.89, P<0.05 or P<0.01). Similar results were observed in fluorescent quantitative RT-PCR test. Conclusion Overexpression of CKLF1 promotes the proliferation of fibroblasts, the secretion of pro-inflammatory cytokines and the expression of osteogenic related target genes, suggesting that CKLF1 might be involved in the pathological ossification of AS.
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Objective To investigate the effects of chemokine like factor 1 (CKLF1) gene on the proliferative activities and osteogenic potentials of hip ligaments of ankylosing spondylitis (AS) in situ and in vitro. Methods Normal and AS hip ligament specimens were collected from 6 patients with femoral neck fracture and 4 AS patients with severe hip deformities. Ligament specimens were exposed to type Ⅱ colla-genase and obtained a single cell suspension, while phase contrast microscopy and anti-vimentin immuno- fluorescence staining (IFC) were applied to observe the cells. The specimens and fibroblasts were divided and cultured in situ and in vitro respectively, and the recombinant adeno-associated virus (rAAV)-lacZ (E. coli beta-galactosidase gene)and rAAV-hCKLF1 (human CKLF1 cDNA cloned in rAAV-lacZ in place of lacZ) were transduced for 21 days. Cell proliferation (cellularity), secretion of pro-inflammatory cytokines, expression of CKLF1 and CCR4 genes were detected by the water-soluble tetrazolium (WST-1) assay and Hoechst 33258 test (DNA content), enzyme-linked immunosorbent assay (ELISA), IFC test and fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCR), respectively. Statistical analysis significance was conducted using the Student's t test and one-way analysis of variance (ANOVA) (LSD) test where appropriate. Results The second passage of normal and AS cells were positive for anti-vimentin, indicating that the cells were fibroblasts. After transducing with rAAV-hCKLF1 for 21 days, cellularity, WST-1 and Hoechst 33258 assays illustrated that CKLF1 gene transfer promoted cell proliferation (compared with the non-viral transduction and lacZ groups, F=6.98, 64.32, 115.91, P<0.05 or P<0.01). Overexpression of CKLF1 gene enhanced the secretion of pro-inflammatory cytokines (interleukin-6 and tumor necrosis factor-alpha) and the expression of bone-specific extracellular matrix proteins (osteopontin and osteocalcin) (F=34.57, 8.89, P<0.05 or P<0.01). Similar results were observed in fluorescent quantitative RT-PCR test. Conclusion Overexpression of CKLF1 promotes the proliferation of fibroblasts, the secretion of pro-inflammatory cytokines and the expression of osteogenic related target genes, suggesting that CKLF1 might be involved in the pathological ossification of AS.
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Background and purpose:Colorectal cancer is one of the most common malignant tumors.Radical operation remains the primary treatment for the disease.Postoperative recurrence and metastases are the main cause for the patient' death.Reoperation for recurrent colorectal cancer is one of the methods to improve survival rate and quality of life for those patients.This study aimed to explore the cause,diagnosis and surgical treatment of recurrent colorectal cancer after operation.Methods:Thirty-five cases of postoperatively recurrent colorectal carcinoma who were treated from 2003 to 2006 at the Department of Surgical Oncology,2th Hospital of Jiaxing were retrospectively analyzed.Results:35 cases of colorectal cancers were re-operated and the overall rate of resection was 63 %,12 of them received radical resection with 4 recurrent rectal cancers and 8 recurrent colon cancers while others were palliatively treated.We followed up these patients from 6 to 36 months and there were 9 cases in 12 cases of radical respected group who were disease free,1 of them had lung metastases,1 liver metastases.While 23 cases in the palliative group,5 were dead,4 had liver metastases and 12 cases were alive.Of the 35 cases,26 cases(70%)tumor recurred within 3 years after first operations while others were recurrent at 1 year.Conclusions:For the patients with recurrent colorectal cancer,surgery remains one of the choices for the treatment.Multimodality treatment should be recommended for the patients at diagnosis.Completed resection of intestine with primary lesion,standard lymphadenectomy and eradicating peritoneal exfoliated cancer cells are the major steps to prevent recurrence of colorectal cancer.