Osteopontin increases hepatocellular carcinoma cell growth in a CD44 dependant manner

AIM:To investigate the role of osteopontin(OPN) and its splice variants in the proliferation of hepatocellular carcinoma(HCC).METHODS:The expression of OPN variants in HCC cell lines as well as HCC tissue samples and nontumour tissue was studied using polymerase chain reaction.OPN variant cDNAs were cloned into a mammalian expression vector allowing both transient expression and the production of stable OPN expressing cell lines.OPN expression was studied in these cells using Western blotting,immunofluoresnce and enzyme linked immunosorbent assay.A CD44 blocking antibody and siRNA targeting of CD44 were used to examine the role of this receptor in the OPN stimulated cell growth observed in culture.Huh-7 cells stably expressing either OPN-A,-B or-C were injected subcutaneously into the flanks of nude mice to observe in vivo tumour growth.Expression of OPN mRNA and protein in these tumours was examined using reverse transcriptionpolymerase chain reaction and immunohistochemistry.RESULTS:OPN is expressed in HCC in 3 forms,the full length OPN-A and 2 splice variants OPN-B and-C.OPN variant expression was noted in HCC tissue as well as cognate surrounding cirrhotic liver tissue.Expression of these OPN variants in the HCC derived cell line Huh-7 resulted in secretion of OPN into the culture medium.Transfer of OPN conditioned media to na ve Huh-7 and HepG2 cells resulted in significant cell growth suggesting that all OPN variants can modulate cell proliferation in a paracrine manner.Furthermore the OPN mediated increase in cellular proliferation was dependent on CD44 as only CD44 positive cell lines responded to OPN conditioned media while siRNA knockdown of CD44 blocked the proliferative effect.OPN expression also increased the proliferation of Huh-7 cells in a subcutaneous nude mouse tumour model,with Huh-7 cells expressing OPN-A showing the greatest proliferative effect.CONCLUSION:This study demonstrates that OPN plays a significant role in the proliferation of HCC through interaction with the cell surface receptor CD44.Modulation of this interaction could represent a novel strategy for the control of HCC.

MicroRNA signatures in chemotherapy resistant esophageal cancer cell lines

AIM:To investigate expression of microRNA(miRNA)and potential targets in chemotherapy resistant esoph-ageal cancer cell lines.METHODS:An in-vitro model of acquired chemotherapy resistance in esophageal adeno-(EAC)and squamous cell carcinoma(ESCC)cells was used,and microRNA expression profiles for cisplatin or 5-fluorouracil(5-FU)resistant variants vs chemotherapy sensitive controls were compared using microarray and quantitative real-time polymerase chain reaction(PCR).The expression of chemotherapy-relevant genes potentially targeted by the dysregulated microRNAs in the chemotherapy resistant variants was also evaluated.RESULTS:Chemotherapy resistant sublines were found to have specific miRNA signatures,and these miRNA signatures were different for the cisplatin vs 5-FU resistant cells from the same tumor cell line,and also for EAC vs ESCC cells with resistance to the same specific chemotherapy agent.Amongst others,miR-27b-3p,miR-193b-3p,miR-192-5p,miR-378 a-3p,miR-125a-5p and miR-18a-3p were dysregulated,consistent with negative posttranscriptional control of KRAS,TYMS,ABCC3,CBL-B and ERBB2 expression via these miRNAs.CONCLUSION:The current study supports the hypothesis that microRNA expression has an impact on chemotherapy resistance in esophageal cancer.

Trabecular Plate Loss and Deteriorating Elastic Modulus of Femoral Trabecular Bone in Intertrochanteric Hip Fractures

Osteoporotic hip fracture is associated with significant trabecular bone loss, which is typically characterized as low bone density by dual-energy X-ray absorptiometry （DXA） and altered microstructure by micro-computed tomography （pCT）. Emerging morphological analysis techniques, e.g. individual trabecula segmentation （ITS）, can provide additional insights into changes in plate-like and rod-like trabeculae, two major micro- structural types serving different roles in determining bone strength. Using ITS, we evaluated trabecular microstructure of intertrochanteric bone cores obtained from 23 patients undergoing hip replacement surgery for intertrochanteric fracture and 22 cadaveric controls. Micro-finite element （~FE） analyses were performed to further understand how the abnormalities seen by ITS might translate into effects on bone strength. ITS analyses revealed that, near fracture site, plate-like trabeculae were seriously depleted in fracture patients, but trabecular rod volume was maintained. Besides, decreased plate area and rod length were observed in fracture patients. Fracture patients also showed decreased elastic moduli and shear moduli of trabecular bone. These results provided evidence that in intertrochanteric hip fracture, preferential loss of plate-like trabeculae led to more rod-like microstructure and deteriorated mechanical competence adjacent to the fracture site, which increased our understanding of the biomechanical pathogenesis of hip fracture in osteoporosis.

Extent and predictors of grade upgrading and downgrading in an Australian cohort according to the new prostate cancer grade groupings

Object:To determine the extent and impact of upgrading and downgrading among men who underwent radical prostatectomy(RP)according to new grade groupings and to identify predictors of upgrading from biopsy grade Group Ⅰ and Ⅱ,and downgrading to grade Group I,in a community setting.Methods:Study participants included 2279 men with non-metastatic prostate cancer diagnosed 2006-2015 who underwent prostatectomy,from the multi-institutional South Australia Prostate Cancer Clinical Outcomes Collaborative registry.Extent of up-or down-grading was assessed by comparing biopsy and prostatectomy grade groupings.Risk of biochemical recurrence(BCR)with upgrading was assessed using multivariable competing risk regression.Binomial logistic regression was used to identify pre-treatment predictors of upgrading from grade Groups Ⅰ and Ⅱ,and risk group reclassification among men with low risk disease.Results:Upgrading occurred in 35%of cases,while downgrading occurred in 13%of cases.Sixty percent with grade Group I disease were upgraded following prostatectomy.Upgrading from grade Group I was associated with greater risk of BCR compared with concordant grading(Hazard ratio:3.1,95%confidence interval:1.7-6.0).Older age,higher prostate-specific antigen levels(PSA),fewer biopsy cores,higher number of positive cores and more recent diagnosis predicted upgrading from grade Group Ⅰ,while higher PSA and clinical stage predicted upgrading from grade Group Ⅱ.No clinical risk factors for reclassification were identified.Conclusion:Biopsy sampling errors may play an important role in upgrading from grade Group I.Improved clinical assessment of grade is needed to encourage greater uptake of active surveillance.

Serotypes of Non-O157 Shigatoxigenic <i>Escherichia coli</i>(STEC)

Non-O157 STEC has been shown to have a diverse ecological distribution among food-animals. It has been associated with both outbreaks and individual cases of severe illness. This group of the organisms is now considered as a major contributor to human disease. The clinical description of the diseases caused by these organisms is reviewed. The host specificity of these pathogens is described and discussed. These organisms appear widespread among food animals like cattle and sheep, and can therefore affect a range of foods directly from the meat and excretions of these animals being used in farming practices. This article reviews the origins, diversity and pathogenesis of non-O157 STEC.

Multinational survey on the preferred approach to management of Barrett’s esophagus in the Asia-Pacific region

BACKGROUND Major societies provide differing guidance on management of Barrett’s esophagus(BE),making standardization challenging.AIM To evaluate the preferred diagnosis and management practices of BE among Asian endoscopists.METHODS Endoscopists from across Asia were invited to participate in an online questionnaire comprising eleven questions regarding diagnosis,surveillance and management of BE.RESULTS Five hundred sixty-nine of 1016(56.0%)respondents completed the survey,with most respondents from Japan(n=310,54.5%)and China(n=129,22.7%).Overall,the preferred endoscopic landmark of the esophagogastric junction was squamocolumnar junction(42.0%).Distal palisade vessels was preferred in Japan(59.0%vs 10.0%,P<0.001)while outside Japan,squamo-columnar junction was preferred(59.5%vs 27.4%,P<0.001).Only 16.3%of respondents used Prague C and M criteria all the time.It was never used by 46.1%of Japanese,whereas 84.2%outside Japan,endoscopists used it to varying extents(P<0.001).Most Asian endoscopists(70.8%)would survey long-segment BE without dysplasia every two years.Adherence to Seattle protocol was poor with only 6.3%always performing it.73.2%of Japanese never did it,compared to 19.3%outside Japan(P<0.001).The most preferred(74.0%)treatment of non-dysplastic BE was proton pump inhibitor only when the patient was symptomatic or had esophagitis.For BE with low-grade dysplasia,6-monthly surveillance was preferred in 61.9%within Japan vs 47.9%outside Japan(P<0.001).CONCLUSION Diagnosis and management of BE varied within Asia,with stark contrast between Japan and outside Japan.Most Asian endoscopists chose squamo-columnar junction to be the landmark for esophagogastric junction,which is incorrect.Most also did not consistently use Prague criteria,and Seattle protocol.Lack of standardization,education and research are possible reasons.

Gastro-oesophageal reflux disease and eosinophilic oesophagitis: What is the relationship?

Eosinophilic oesophagitis(EoE) and gastro-oesophageal reflux disease(GORD) are the most common causes of chronic oesophagitis and dysphagia associated with oesophageal mucosal eosinophilia. Distinguishing between the two is imperative but challenging due to overlapping clinical and histological features. A diagnosis of EoE requires clinical, histological and endoscopic correlation whereas a diagnosis of GORD is mainly clinical without the need for other investigations. Both entities may exhibit oesophageal eosinophilia at a similar level making a histological distinction between them difficult. Although the term proton-pump inhibitor responsive oesophageal eosinophilia has recently been retracted from the guidelines, a relationship between Eo E and GORD still exists. This relationship is complex as they may coexist, either interacting bidirectionally or are unrelated. This review aims to outline the differences and potential relationship between the two conditions, with specific focus on histology, immunology, pathogenesis and treatment.

阿哌沙班vs依诺肝素用于膝关节置换术后血栓预防(ADVANCE-2):随机双盲试验

背景依诺肝素等低分子肝素是预防大关节置换术后静脉血栓形成的首选。阿哌沙班是一种口服活性的Xa因了抑制剂,在预防大关节置换术后静脉血栓形成方面可能有效,而且相对于依诺肝素来说,出血风险较低、用法也较简便。本研究评估选择性全膝关节置换术后卜述药物的有效性和安全性。方法 ADVANCE-2为多中心随机双盲Ⅲ期研究,选择性双侧或单侧全膝关节置换术患者通过交互式中心电话系统进行分组,随机接受阿哌沙班2.5mg每天2次口服(1528例)或依诺肝素40mg每天1次皮下注射(1529例)。随机化方案由Bristol-Myers Squibb随机中心生成,按研究中心和单侧或双侧手术分层,区组大小为4。研究者、患者、统计专家、评判者和指导委员会不知道患者分组情况。阿哌沙班于手术伤口闭合后10～12小时使用,依诺肝素于术前12小时使用;两药均持续用药10-14天,按照计划用药结束时行双侧顺行性静脉造影。主要终点是治疗期间无症状和有症状的深静脉血栓形成、非致死性肺栓塞和全因死亡的复合指标。统计学计划要求在检验阿哌沙班的优效性之前达到非劣效;非劣效检验对意向治疗患者进行分析。本研究在ClinicalTrials.gov备案,备案号NCT00452530。结果接受治疗的3057例患者(阿哌沙班组1528例、依诺肝素组1529例)中,1973例符合主要有效性分析的要求。阿哌沙班组976例和依诺肝素组997例中,报告主要终点的患者分别为147例(15%)和243例(24%)[相对危险度0.62(95%可信区间CI为:0.51-0.74),P<0.0001;绝对危险度下降9.3%(5.8～12.7)]。阿哌沙班组1501例和依诺肝素组1508例中,发生大出血或临床相关非大出血的患者分别为53例(4%)和72例(5%)(P=0.09)。解读全膝关节置换术后次日上午使用阿哌沙班2.5mg每天2次,是依诺肝素40mg/天简便而更有效的口服替代措施,且不会增加出血。