Establishment of methylation detection method for Septin 9 gene and its application in plasma specimen detection in colorectal cancer / 国际检验医学杂志

Objective To establish the blood Septin9 methylation detection system for early screening of colorectal cancer based on fluorescence PCR technology .Methods The PCR primer of Septin9 was designed by searching the CpG island site of Septin9 methylation in the NCBI database .The high methylation site of Septin9 gene promoter region was confirmed by PCR amplification and sequencing after extracting DNA from colorectal cancer and para-carcinoma tissues .The fluorescence PCR and TaqMan probe detection technique was designed by aiming at high methylation site for constructing the plasma sample methylation detection sys-tem .Then its accuracy ,specificity ,repeatability and minimum detectable amount were performed the assess-ment and analysis .The SETP9 methylation detection was performed in plasma samples from 57 cases of color-ectal cancer and 30 healthy persons .Results The high methylation site of Septin9 gene in tissue samples was confirmed by sequencing .This site served as the target for designing fluorescence PCR detection system .After assessment ,the accuracy ,specificity and repeatability of this detection system were 100％ ,the lowest detection amount reached 0 .1 ng/μL .Among the plasma samples in 57 cases of colorectal cancer ,the positive rate of Septin9 methylation site detection was 71 .92％ (41/57) and the positive rate of in the patients with pathologi-cal stage Ⅰ and Ⅱ of colorectal cancer reached 64 .2％ .But Septin9 gene had no methylation in plasma of healthy population .Conclusion The plasma fluorescence PCR detection system with Septin9 gene methylation as the target has the characteristics of high sensitivity ,high specificity and high accuracy ,which is the reliable detection technique for the early screening of colorectal cancer and has good clinical application prospect .

Expression difference of DNA mismatch repair gene hMLH1 and hMSH2 between schistosomiasis-associated colorectal cancer and sporadic colorectal cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the expression difference of DNA mismatch repair gene hMLH1 and hMSH2 between schistosomiasis-associated colorectal cancer and sporadic colorectal cancer.</p><p><b>METHOD</b>Clinical and pathological data of colorectal cancer patients receiving operations in Zhejiang Cancer Hospital between January 2008 and December 2010 were retrospectively analyzed. Patients were divided into schistosomiasis group(n=80) and sporadic group (n=258) according to the preoperative history and pathologic results. Pathological specimens were collected and tissue chips were made to analyze the expression of hMLH1 and hMSH2 by immunohistochemistr.</p><p><b>RESULTS</b>Compared with sporadic group, older age [(62.2 ± 9.6) year vs. (57.2 ± 11.7) year, P=0.000)], lower platelet level [(197.0 ± 59.6) × 10(9)/L vs. (217.0 ± 84.3) × 10(9)/L, P=0.02] and lower WBC level [(5.9 ± 1.9) × 10(9)/L vs. (6.6 ± 2.8) × 10(9)/L, P=0.02] were found in schistosomiasis group. Ratio of low differentiation-undifferentiation tumor was significantly higher in schistosomiasis group [44.2% (34/77) vs. 4.9% (12/247), P<0.05]. Lower positive rate of hMLH1 expression [77.5% (62/80) vs. 98.1% (253/258), P=0.000] and hMSH2 expression [75.0% (60/80) vs. 95.3% (246/258), P=0.000] was found in schistosomiasis group compared with sporadic group. Concurrent schistosomiasis was one of the risk factors of hMLH1/hMSH2 deficiency (RR: 0.913, 95% CI: 0.836-0.997, P=0.043), but not an independent factor (RR: 0.951, 95% CI: 0.867-1.043, P=0.286).</p><p><b>CONCLUSION</b>Schistosomiasis is associated with lower positive expression of hMLH1 and hMSH2, which indicates that hMLH1/hMSH2 deficiency may be a potential mechanism of schistosomiasis inducing carcinogenesis of colorectal cancer.</p>

Expressions of P53,topoisomerase Ⅱ and multi-drug resistance associated protein in tissues of colorectal cancer of patients combined with chronic schistosomiasis / 中华消化外科杂志

Objective To investigate the expressions of P53,topoisomeraseⅡ(TopoⅡ)and multidrug resistance associated protein(MRP)in tissues of colorectal cancer of patients combined with chronic schistosomiasis.Method The retrospective case-control study was adopted.The clinicopathological data of 338 colorectal cancer patients who were admitted to the Zhejiang Cancer Hospital between January 2008 and December 2010 were collected.Cancer tissue specimens from surgical resection were collected.Among 338 patients,80 were combined with chronic schistosomiasis and 258 were combined with non-chronic schistosomiasis.The expressions of P53,TopoⅡand MRP were dectected using immunohistochemistry(IHC).Ranked data were presented as percentage and analyzed using the non-parametric test.Results The negative,weak positive,positive and strong positive expressions of P53 were respectively 5.00％(4/80),87.50％(70/80),3.75％(3/80),3.75％(3/80)in tissues of colorectal cancer of patients combined with chronic schistosomiasis and 28.68％(74/258),19.38％(50/258),16.67％(43/258),35.27％(91/258)in tissues of colorectal cancer of patients combined with non-chronic schistosomiasis,with a statistically significant difference(Z=-2.962,P＜0.05).The negative,weak positive,positive and strong positive expressions of TopoⅡwere respectively 8.75％(7/80),51.25％(41/80),22.50％(18/80),17.50％(14/80)in tissues of colorectal cancer of patients combined with chronic schistosomiasis and 12.01％(31/258),55.43％(143/258),22.48％(58/258),10.08％(26/258)in tissues of colorectal cancer of patients combined with non-chronic schistosomiasis,with no statistically significant difference(Z=-1.551,P＞0.05).The negative,weak positive,positive and strong positive expressions of MRP were respectively 7.50％(6/80),40.00％(32/80),28.75％(23/80),23.75％(19/80)in issues of colorectal cancer of patients combined with chronic schistosomiasis and 24.42％(63/258),38.37％(99/258),24.03％(62/258),13.18％(34/258)in tissues of colorectal cancer of patients combined with non-chronic schistosomiasis,with a statistically significant difference(Z=-3.408,P＜0.05).Conclusion There are abnormal expressions of P53 and MRP in tissues of colorectal cancer of patients combined with chronic schistosomiasis,which may be involved in the hypothetical mechanism of chronic schistosomiasis inducing carcinogenesis of colorectal cancer.

The anti-neuroinflammatory effects of dehydromiltirone and related mechanisms / 中国药理学通报

Aim To investigate the anti-neuroinflam-matory activities of dehydromiltirone and the underlying mechanisms in LPS-stimulated microglial cell line BV2 cells. Methods BV2 cells were pre-treated with de-hydromiltirone, then stimulated by LPS. The levels of nitric oxide( NO) were measured by Griess assay, and the concentrations of pro-inflammatory cytokines were measured by ELISA assay. Confocal fluorescence mi-croscopy was used to measure the expression of MAC-1, the biomarker of activated BV2 cells. The levels of-inducible nitric oxide synthase ( iNOS ) , cyclooxygen-ase-2 ( COX-2 ) , NF-κB and PI3 K/Akt were deter-mined by Western blot analysis. Results The treat-ment of dehydromiltirone significantly inhibited the pro-duction of NO, TNF-α and IL-6, attenuated the ex-pression of iNOS and COX-2 protein, and dampened the microglial activation in LPS-stimulated BV2 cells. The mechanistic study revealed that dehydromiltirone inhibited the phosphorylation of PI3 K and Akt in LPS-stimulated BV2 cells, and decreased NF-κB activation by suppressing the degradation of IκB. Conclusion dehydromiltirone shows significant anti-neuroinflamma-tory effects through inhibiting PI3 K/Akt phosphoryla-tion and then inhibiting NF-κB signaling pathway.

Research progress in the study of protective effect of tanshinone IIA on cerebral ischemic stroke / 药学学报

Danshen is one of the traditional Chinese herbal medicines and nas a long history or being used clinically in the treatment of cardiovascular and cerebrovascular conditions such as coronary heart disease and angina pectoris. Tanshinone IIA is a derivative of phenanthrene-quinone isolated from Danshen. It has been reported to be the major bioactive compound of Danshen and has diverse biological effects. Recent studies demonstrated that tanshinone IIA had neuroprotective effects on experimental ischemic stroke through its antiinflammatory, anti-oxidant, anti-apoptosis effects and its inhibitory effect on excitatory amino acid toxicity. In this review, we summarized all the recent progresses on the protective effect of tanshinone IIA on cerebral ischemic stroke. Hopefully, this article will throw some light on further study and application of tanshinone IIA.

Research progress of gastrointestinal stromal tumor / 国际肿瘤学杂志

Gastrointestinal stromal tumor is one kind of the most common gastrointestinal tumors derived from the mesenchymal tissue.In most cases,it occurs in adults,while rarely occurs in adolescents.The clinical diagnosis of pediatric GIST can refer to the diagnosis standard of adults GIST,but it pays more attention to genetic diagnosis.The main treatment for pediatric GIST is surgery,but for advanced or unresectable patients,we can adopt imatinib or sutent for targeted therapy according to the results of genetic testing.

Comparison of short-term outcomes of hand-assisted laparoscopic, laparoscopic, and open surgery in the treatment of rectal cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>The aim of this study was to compare the short-term outcomes for hand-assisted, laparoscopic, and open resection for rectal cancer.</p><p><b>METHODS</b>Three hundred ninety patients with rectal cancer who underwent curative resection between June 2009 and June 2012 were included. Patients were classified into a hand-assisted group (HALS, n=101), a laparoscopic surgery group (LS, n=157), and an open surgery group (OS, n=132). Patient and disease characteristics, operative parameters, postoperative morbidity, pathological results and length of recovery were compared among three groups.</p><p><b>RESULTS</b>The mean operating time was (173±39) min for the HALS group, (231±61) min for the LS group, and (173±39) min for the OS group (P<0.01). Conversion rates did not differ between HALS and LS groups (2.0% vs 3.2%, P=0.708). The overall complication rates were 11.9%, 11.5%, and 19.7% in the HALS, LS and OS groups respectively (P=0.100). The specimen quality with a specimen length, distal resection margin, harvested lymph nodes, and positive lymph nodes did not differ among the three groups. Patients in the HALS and LS groups recovered significantly faster than those from the OS group.</p><p><b>CONCLUSIONS</b>This comparative study shows that HALS and LS can reproduce the equivalent short-term results of standard OS. HALS retained the minimal invasive advantages of LS, and significantly shorten the operation time.</p>

Formulation and Evaluation of Irinotecan Suppository for Rectal Administration

Irinotecan suppository was prepared using the moulding method with a homogeneous blend. A sensitive and specific fluorescence method was developed and validated for the determination of irinotecan in plasma using HPLC. The pharmacokinetics of intravenous administered and rectal administered in rabbits was investigated. Following a single intravenous dose of irinotecan (50 mg/kg), the plasma irinotecan concentration demonstrated a bi-exponential decay, with a rapid decline over 15 min. C(max), t(1/2), AUC(0-30h) and AUC(0-infinity) were 16.1 +/- 2.7 g/ml, 7.6 +/- 1.2 h, 71.3 +/- 8.8 microg.h/ml and 82.3 +/- 9.5 microg.h/ml, respectively. Following rectal administration of 100 mg/kg irinotecan, the plasma irinotecan concentration reached a peak of 5.3 +/- 2.5 microg/ml at 4 h. The AUC(0-30h) and AUC(0-infinity) were 32.2 +/- 6.2 microg.h/ml and 41.6 +/- 7.2 microg.h/ml, respectively. It representing ~50.6% of the absolute bioavailability.

Formulation and Evaluation of Irinotecan Suppository for Rectal Administration

Irinotecan suppository was prepared using the moulding method with a homogeneous blend. A sensitive and specific fluorescence method was developed and validated for the determination of irinotecan in plasma using HPLC. The pharmacokinetics of intravenous administered and rectal administered in rabbits was investigated. Following a single intravenous dose of irinotecan (50 mg/kg), the plasma irinotecan concentration demonstrated a bi-exponential decay, with a rapid decline over 15 min. C(max), t(1/2), AUC(0-30h) and AUC(0-infinity) were 16.1 +/- 2.7 g/ml, 7.6 +/- 1.2 h, 71.3 +/- 8.8 microg.h/ml and 82.3 +/- 9.5 microg.h/ml, respectively. Following rectal administration of 100 mg/kg irinotecan, the plasma irinotecan concentration reached a peak of 5.3 +/- 2.5 microg/ml at 4 h. The AUC(0-30h) and AUC(0-infinity) were 32.2 +/- 6.2 microg.h/ml and 41.6 +/- 7.2 microg.h/ml, respectively. It representing ~50.6% of the absolute bioavailability.

Data analysis of 256 consultations of patients with ostomy by telephone / 中华护理杂志

Objective To analyze the ostomy patients' demands, resolve the common problems after discharge by telephone consultation, and supplement the content of specialized health education. Methods The data of 256 telephone consultations from 186 patients with ostomy were retrospectively analyzed. Results Most of the consultations came from patients aged from 61 to 80 years old (64.5%) and patients within one year after surgery (85%). The contents focused on information about ostomy products, care of skin around the stoma and leakage of defecation, which accounted to 32.8%,30.1%,24.6%,respectively.The problems were solved by telephone interview in 201 consultations (78.5%). Conclusion Consultation by telephone is useful to find out the demands of discharged ostomy patients, resolve their problems in time and improve their quality of life. It has great significance for improving health education and extensive services of ostomy patients.

Multiple primary colorectal carcinoma / 中华普通外科杂志

Objective To investigate the diagnosis and surgical therapy of multiple primary colorectal carcinoma. Methods From 1998 to 2007, 47 patients with synchronous multiple primary colorectal carcinoma and 20 cases with metachronous carcinoma were treated in our hospital. Results In these 67 cases of multiple primary colorectal carcinoma, synchronous carcinoma (SC) accounted for 70% (47 cases) including 37 rectal cancer with a total of 95 larger bowel cancer lesions. There were 6 cases with Dukes A stage, 22 cases with Dukes B stage, 15 cases with Dukes C stage and 4 cases with Dukes D stage. In this whole group there were 20 cases with lymph node metastasis, 21 cases with adenoma and multiple polyps in SC. Three cases received total coloectomy, 10 cases did subtotal coloectomy, 34 cases were treated by radical resection and intestine segment resection. In 20 metachronous carcinoma cases, there were 31 colon cancer(70%) with a total of 44 intestinal cancer lesions. Altogether, there were 17 cases with two tumors, 2 cases with three tumors, one case with four tumors. The duration between the first and the last carcinoma was from 7 months to 19 years, including less than two years in 7 cases, from two to five years in 5 cases, and more than five years in 8 cases. In all 20 MC cases the first (primary) carcinoma received radical resection, while radical resection was performed for the secondary carcinoma in 14 cases and for the third carcinoma in 2 cases. In the SC and the primary carcinoma of MC patients who received radical resection, the 5-year survival rates were 74% and 78% respectively. Conclusion In cases of colonic carcinoma we shouldn't be satisfied with the diagnosis of single colon tumor before a thorough screening of the whole colon was made. In radical resection surgery for SC or MC cases an attempt to preserve enough residual intestinal tract should be made in order to improve the life quality of post-operative patients.

Expression and clinical significance of guanylyl cyclase C mRNA in peripheral blood of the patients with colorectal cancer / 国际外科学杂志

Objective To investigate the expression of guanylyl eyelasec(GC-C)mRNA in pefipheral blood of colorectal cancer patients and discuss its clinical significance. Methods To detect the GC-C mRNA in peripheral blood of colorectal cancer patients by reverse transcriptase PCR and analysis their clinical and pathological indexes. Results The positive rate of GC-C mRNA in 52 colorectal cancer patients was 50%(26/52),while to patients with extra-intestinal cancer,the positive rate was 0(0/8).There were significant difierences of positive rate of GC-C mRNA associated with tumor infiltration(P＜0.05)and tumor embolus in vessel(P＜0.05).While to CEA level in peripheral blood(P＞0.05),but there were no statistical difference.The positive rate of micrometastasis in coloreetal cancer patients might increase using the GC-C mRNA combined with CEA level in peripheral bood.The positive rate of GC-C mRNA in colorectal cancer patients with Dukes A、B、C and D stages were 0(0/3),47.06%(8/17),52.38%(11/21),63.63%(7/11),The expression of GC-C mRNA increased with the clinical stage chang,there was no statistical difference.Conclusions GC-C expresses selectively in peripheral blood of colorectal cancer patients.Itis a valuable new tumor marker in micrometastasis diagnosis and post-operation follow-up of colorectal cancer,and it can also be used as the location determination of metastasis intestinal tumor.

Detection of COX-2 and its relationship with MVD in colorectal carcinoma tissue / 肿瘤研究与临床

Objective To investigate the expression of COX-2 and angiogenesis in colorectal carcinoma tissue.Methods The expressions of COX-2 and MVD of 88 cases of colorectal carcinoma and 20cases of control tissue were examined by immunohistochemical staining. Results Compared with normal control,positive expression rates of COX-2(3/20 vs 64/88),and CD34(7/20 vs 74/88)of colorectal carcinoma tissue increased significantly(P＜0.05).MVD in COX-2 positive expressed tissue(63.24-20.4)was higher than that in COX-2 negative expressed tissue(41.2±29.8)(P＜0.01).Conclusion COX-2 is overexpressed in colorectal carcinoma tissue,which may have a strong relationship with MVD.

Comparative study of transrectal ultrasonography and spiral computerized tomography in preoperative staging of rectal cancer:contrasted with the pathologic findings / 中华超声影像学杂志

Objective To compare the efficacy of transrectal ultrasonography(TRUS)and spiral computed tomography(SCT)in preoperative staging of rectal carcinoma contrasted with the postoperative pathologic findings.Methods Both TRUS and SCT were performed prior to surgery in 92 patients with rectal carcinoma.After radical operation,the preoperative findings were compared with the histological findings,and the the efficacy of TRUS and SCT in staging the rectal carcinoma were evaluated.Results The accuracy of TRUS for T stage and N stage was 87.0% and 64.1% respectively,while the accuracy of SCT was 68.5% and 66.3%.Conclusions TRUS is superior to SCT for the judgment of tumor infiltration depth,TRUS may become the first choice in preoperative staging of rectal carcinoma.But neither is able to provide satisfaction assessment for lymph node metastases.When both methods are used together,it would be better.

Laparoscopic Total Mesorectal Excision in Treatment of Mid-Low Rectal Cancer(Report of 74 Cases) / 中国普外基础与临床杂志

Objective To evaluate the feasibility and clinical outcomes of laparoscopic total mesorectal excision(TME) in treating mid-low rectal cancer.Methods From March 2005 to July 2008,74 patients with mid-low rectal cancer undergoing laparoscopic TME in Zhejiang Cancer Hospital were collected.The data of clinicopathologic parameters were analyzed.Results Laparoscopic TME was performed on 74 patients with mid-low rectal cancer.No operative death occurred in this group.No case was converted to open procedure.The mean operation time was 187 min.The mean operative blood loss was 90 ml.The mean postoperative hospital stay was 10 d.Bowel function was restored on 46 h after operation on average.The mean distance between tumor and the section edge was 3.1 cm.The average number of lymph node dissection was 19.7.The sphincter preservation rate was 97% in patients with tumor 6 cm above the anal verge.The follow-up times were 2-44 months,average 25 months.The incidence of complications was 9.5%.No tumor cell port site implantation or distant metastasis happened.One case was pelvic recurrence,no patient was dead.Conclusion Laparoscopic TME is a feasible,safe and minimally invasive technique for the patients with mid-low rectal cancer,achieving the principles of TME.