人胆囊癌细胞系GBC-SD中侧群细胞的耐药性研究

目的 研究人胆囊细胞系GBC-SD中侧群细胞(side population cells,SP)的耐药特性,并探讨其耐药机制.方法 利用流式细胞术分选SP、非SP细胞,采用MTT检测2种细胞亚群对5种化疗药物吉西他滨、顺铂、5-氟尿嘧啶、表阿霉素、米托蒽醌的药物敏感性;利用流式细胞术检测吉西他滨处理的人胆囊癌细胞系GBC-SD中SP细胞比例的变化,并通过RT-PCR、Western印迹检测SP、非SP细胞亚群中ABCG2 mRNA和蛋白的表达.结果 吉西他滨、顺铂、5-氟尿嘧啶、米托蒽醌以对胆囊癌细胞系GBC-SD的IC50浓度分别作用SP、非SP细胞亚群1 d后,SP细胞的增殖能力高于非SP细胞,两组之间的差异具有统计学意义(P<0.05),而表阿霉素处理水平的两组间差异无统计学意义(P>0.05);人胆囊癌细胞系GBC-SD经过吉西他滨处理3周后,SP细胞的比例显著升高(8.02%±0.13%比0.62%±0.08%,P<0.05),且SP细胞明显高表达ABCG2基因.结论 人胆囊癌细胞系GBC-SD中SP细胞具有类似干细胞的耐药特性,耐药基因ABCG2高表达是其耐药的重要机制.

Abstract：

Objective To investigate the drug resistance of side population cells in human gallbladder cancer cell line GBC-SD and explore its mechanism. Methods Drug sensitivity assays of 5chemotherapeutic agents were performed on side population cells (SP) and non-SP cells of GBC-SD.GBC-SD was cultured and then treated with the chemotherapeutic agent gemcitabine. The frequency of SP by FACS was measured. RT-PCR and Western blotting were used to detect the expression of AB-CG2 in both the SP and the corresponding non-SP subsets. Results After 1 d treatment with 4 chemotherapeutic agents (gemcitabine, cisplatin, 5-fluorouracil and mitoxantrone) in IC50 concentration to GBC-SD cell line, the reproductive ability of SP was higher than that of non-SP (P<0.05). However, statistical significance was not achieved when compared with epirubicin (P>0.05). The percentage of SP in GBC-SD treated with chemotherapeutic agent gemcitabine after 3 weeks was sharply elevated by FACS (8.02% ±0.13% vs 0.62% ±0.08%, P<0.05), and the expression of ABCG2mRNA and protein were increased in SP as compared with non-SP. Conclusion SP from human gallbladder cancer cell line GBC-SD, like stem cell, showed a heighten resistance to drugs. Increased expression of ABCG2 was largely responsible for the multi-drug resistance.     

苏皖地区汉族人群IL-6-572C>G多态性与前列腺癌易感性的关系

目的:研究中国苏皖地区汉族人群IL-6-572C>G多态性与前列腺癌易感性的关系。方法:采用病例-对照研究,提取200例前列腺癌患者(病例组)和279例健康体检的非肿瘤患者(对照组)外周血基因组DNA,应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析两组IL-6-572C>G位点的多态性,比较不同基因型与前列腺癌易感性的关系。结果:以携带CC基因型的对象为参照组,携带GG基因型者患前列腺癌的风险提高146%(OR=2.46,95%C I:1.41～4.29)。而携带GG基因型者患前列腺癌的风险是携带GC/CC基因型的2.47倍(95%C I:1.47～4.17)。在高龄(>70岁)、高体重指数(>23 kg/m2)、非吸烟、饮酒和有肿瘤家族史5组人群中,携带GG基因型的个体发病风险显著升高,调整后的OR(95%C I)分别为3.06(1.44～6.49)、3.72(1.79～7.74)、2.96(1.30～6.72)、2.73(1.28～5.79)、6.67(1.50～29.69)。结论:IL-6-572C>G与苏皖地区汉族人群前列腺癌的易感性有关,GG基因型可能是前列腺癌的易感基因型。     

产后小鼠子宫内膜边缘群干/祖细胞的分离纯化

目的分离和纯化小鼠子宫内膜边缘群(side population,SP)干/祖细胞,为进一步探讨产后子宫修复的细胞机制奠定基础。方法分别采用酶消化和机械分离结合法与机械研磨分离的方法,原代分离小鼠子宫内膜上皮细胞与基质细胞,通过Hoechst33342-SP法(对照加入维拉帕米),使用流式细胞仪检测子宫内膜SP细胞。结果未孕小鼠子宫内膜上皮与基质中均无明确SP干/祖细胞,产后哺乳期后的小鼠子宫内膜基质中含有SP干/祖细胞达(3.44±1.59)%。结论本研究初步检测到小鼠产后子宫内膜中含有SP干/祖细胞。产后更多SP细胞的出现可能与修复损伤的子宫内膜有关。这群SP细胞是子宫原位干细胞还是有其他来源,如何参与子宫内膜的修复,其分子机制如何,尚待进一步研究。     

Liver Transplantation for Cystic Fibrosis in Adults

Purpose To expand our knowledge on liver transplantation for cirrhosis associated with cystic fibrosis in adults. Methods Five patients who underwent a liver transplantation due to cystic fibrosis were reviewed. The outcome of the patients in terms of age, immunosuppression regimen, patient and graft survival, and pre- and post-transplant complications were investigated. Results Five adult liver transplant patients had cystic fibrosis (0.2%). These included 4 men and 1 woman with a mean age of 31 ± 10, ranging from 22 to 52 years old at the time of transplantation. All patients had lung problems. Four patients had exocrine and two had endocrine pancreatic insufficiency. Two are currently alive with a follow-up of 5.8 years and 4 months after transplantation, respectively. There were three deaths from pulmonary embolism at 4.5 years, myocardial infarction with cyclosporine nephrotoxicity at 10.7 years, and lymphoproliferative disorder at 5 months after transplantation. No deaths occurred from lung infection. Only one patient had postoperative pulmonary infectious complications, which were successfully treated with antibiotics and did not result in mortality. Conclusion Adult liver transplantation for end-stage liver disease associated with cystic fibrosis offers encouraging results with a rapid general improvement after surgery and it is now considered to be a safe and acceptable treatment for this disease population.     

SURVEILLANCE CAN BE THE STANDARD OF CARE FOR STAGE I NONSEMINOMATOUS TESTICULAR TUMORS AND EVEN HIGH RISK PATIENTS

PURPOSE: We investigate the results of a surveillance program for stage I nonseminomatous germ cell tumors to validate a surveillance policy, and furthermore improve it by analyzing diagnostic instruments and identifying prognostic factors for relapse. MATERIALS AND METHODS: From 1982 to 1994, 90 patients with stage I nonseminomatous germ cell tumors entered a surveillance protocol after orchiectomy. Patients with relapse were treated with cisplatin based chemotherapy. A statistical analysis of possible prognostic factors for relapse was performed. RESULTS: Relapse occurred in 23 (26%) patients. Disease specific survival was 98.9%, and 1 patient died of tumor. Most relapses were located in retroperitoneal lymph nodes only (78%). Tumor markers were the most important indicators of relapse. However, in 22% of patients with relapse abdominal x-ray of lymphangiographic contrast showed the first sign of relapse. Computerized tomography located all but 1 relapse. Vascular invasion (p = 0.0001), tumor size (p = 0.0341) and presence of immature teratoma (p = 0.0154) were significantly predictive of relapse with the multivariate analysis, percentage embryonal carcinoma only by univariate analysis (p = 0.032). The relapse rate was highest (52%) when vascular invasion was present. CONCLUSIONS: With surveillance for stage I nonseminomatous germ cell tumors, excellent treatment results can be achieved that are comparable to primary retroperitoneal lymph node dissection. Tumor markers and computerized tomography are highly reliable for detecting relapse. Lymphangiography is still of staging value. Pathological factors may influence the choice of adjuvant treatment. However, relapse risks of 50% to 60% are maximally achieved with presently available prognostic factors, and so sparing morbidity of adjuvant treatment by a surveillance protocol remains a feasible option even in these patients.     

Risk‐stratified intensive follow up for treated colorectal cancer – realistic and cost saving?

Objective Intensive follow‐up post surgery for colorectal cancer (CRC) is thought to improve long‐term survival principally through the earlier detection of recurrent disease. This paper aims to calculate the additional resource and cost implications of intensive follow up post‐CRC resection, examine the possibility of risk‐stratifying this follow up to those at highest risk of recurrence and investigating the impact that population screening might have on the future cost and outcomes of follow up. Method Two follow‐up regimens were constructed: the ‘standard’ follow‐up protocol used the principles of the British Society of Gastroenterology (BSG) guidelines whilst the ‘intensive’ follow‐up protocol used the most intensive arm of the follow up after colorectal surgery (FACS) trial. Using ONS data, the number of CRC diagnosed in a given year was calculated for 2003 and projected for 2016 based on the population of England and Wales. The resource requirements and costs of follow up over a 5‐year period were then calculated for the two time periods. Risk stratifying entry to follow up and the introduction of population CRC screening were then considered. Results For the 2003 cohort, an intensive follow‐up program would detect 853 additional resectable recurrences over 5 years with 795 fewer subjects requiring palliative care. An additional 26 302 outpatient appointments, 181 352 CEA tests and 79 695 CT scans over 5 years would be required to achieve this. The cost of investigating subjects who would never develop detectable recurrences was £15.6 million. The cost per additional resectable recurrence was £18 077, a figure also found for a nonscreened population in 2016. An identical intensive follow‐up policy with biennial FOBT screening in 2016 saw the cost per additional resectable recurrence rise to £36 255. Conclusion Intensive follow up will detect considerably more resectable recurrences but at considerable cost and it is unclear if such follow up will be achievable in an already over‐stretched NHS. If population‐based CRC screening increases the number of Dukes A cancers this may offer the possibility of risk‐stratifying future follow up to those at highest risk of recurrence; minimizing tests on those who will never have recurrent disease and better utilizing our scarce resources.     

Patient comprehension and recall of laparoscopic surgery and outcomes in a non-English speaking population.

BACKGROUND AND OBJECTIVES: The purpose of this study was to determine patient recall and comprehension after laparoscopic appendectomy in an underserved population. Laparoscopic surgery can lead to diagnostic uncertainty secondary to poor recall and variable port placement. METHODS: After institutional review board approval, we identified a cohort of patients who underwent laparoscopic appendectomy from 2000 to 2004 at a single institution. We then attempted to contact the patients to conduct a 10-question telephone survey, which determined whether the patient spoke English or Spanish as a primary language, ethnicity, educational level, and questions about recall of perioperative events and diagnoses. If we could not reach the patient, we tried to call back on 2 different occasions. RESULTS: Between 2000 and 2004, 186 patients underwent laparoscopic appendectomy. Of these, 65% were Hispanic. We found that only 17% of these patients returned for a postoperative visit. Only 19.3% could be contacted by phone. Forty-seven percent of the patients contacted by phone spoke Spanish exclusively. Overall 92% of patients contacted knew what operation they had, and gave their correct diagnosis. CONCLUSIONS: The low percentage of patients available to follow-up makes this study statistically insignificant. However, we believe that fact in itself is important. In Southwestern states, we see a large migrant population. This highlights the need to communicate effectively with the patients at the time of surgery, which we speculate we did based on the percentage of patients that knew their diagnosis.     

Complications in patients of acetabular fractures and the factors affecting the quality of reduction in surgically treated cases

IntroductionAcetabulum fractures, though relatively uncommon, are associated with significant morbidity and mortality. These involve high energy trauma and due to their complex nature, the management requires understanding the relevant surgical anatomy, defining the injury via appropriate radiographic assessment and determining a suitable treatment plan. Literature is scarce for the demographic data, fracture patterns, associated injuries, management and early complications in the Indian scenario. These factors play a pivotal role in the ultimate recovery of the patients. Therefore this study was conceptualised to assess the epidemiology and evaluate the complications of these fractures. Furthermore the effects of various factors determining the quality of reduction in surgically treated patients were also assessed.MethodologyThis was a prospective observational study in which patients presenting to the advance trauma centre of our institute with acetabulum fractures were included. Demographical data of the patients, mechanisms of injuries, fractures morphologies, complications and radiological outcomes were recorded prospectively.Results116 patients with acetabular fractures were included in the study. 81% of these were males, with average age of 39.95 ± 15.87 years; with road traffic accidents being the predominant mode of injuries. Mortality was reported in 5 patients; 4 patients had deep venous thrombosis and sciatic nerve injuries were seen in 12 patients of which 4 were iatrogenic. 8 patients had some form of infection, out of which 4 required multiple debridements. 4 cases developed heterotrophic ossification while 2 cases had loss of reduction. The timing of surgery and other associated fractures had significant effect on the quality of reduction (p < 0.05); while age, gender, mode of injury or individual fracture patterns had no such effect.ConclusionsProper radiological assessment and evaluation of fracture configuration is important for management of acetabulum fractures. When indicated, this should be followed by early open reduction and internal fixation to achieve anatomical reduction, with management of associated injuries for better outcomes.     

Post‐mastectomy radiation in node‐positive breast cancer in Ontario

More recent guidelines are more supportive for post‐mastectomy radiation in all node‐positive breast cancer patients. We examined the rate and predictors of post‐mastectomy radiation receipt in Ontario Canada from 2010 to 2014. Of 6535 node‐positive post‐mastectomy patients, 73.9% received radiation. The rate was 68.7% (2903/4227) among women with 1‐3 positive nodes. Radiation was less likely to be administered to women who were older, had high levels of comorbidity, or presented with early stages of breast cancer. Regional practice variation was reassuringly modest.     

Long-term Clinical Significance of Tacrolimus Trough Level at the Early Period After Kidney Transplantation

BackgroundThe stable immunosuppressant level at the early period after kidney transplantation (KT) is one of the most important factors for the prognosis of KT. However, the extent of immunosuppression varies according to the policies of each KT center. We investigated the relationship between the clinical outcome and tacrolimus trough level (TTL) at the early post-transplant period.Materials and MethodsWe retrospectively analyzed medical records of patients who underwent KT between July 2007 and June 2016. We investigated TTLs at 3 months after KT. We evaluated the incidence of biopsy-proven acute rejection (BPAR), cytomegalovirus infection, and graft survival according to the TTLs.ResultsA total of 426 patients who received KT during the study period were enrolled. The mean age of KT recipients was 46.3 ± 11.5 years, and 55.5% of patients were men. The incidence of BPAR within 1 year after KT was significantly higher when TTLs at 3 months were less than 4.0 ng/mL (P = .020). Death-censored graft survival rates were significantly lower in KT recipients with BPAR and TTL less than 4.0 ng/mL (P < .001, P < .001, respectively). In multivariate analysis, BPAR and TTL less than 4.0 ng/mL at 3 months after KT were independent risk factors for graft failure.ConclusionBPAR and TTL less than 4.0 ng/mL at 3 months after KT are important risk factors for allograft failure. Therefore, TTL should be kept at least 4.0 ng/mL or more at 3 months after KT to reduce the incidence of BPAR within 1 year after KT.