移植于裸鼠的人鼻咽癌细胞DNA合成生物节律的初步研究

目的:探索鼻咽癌细胞DNA合成的生物节律为临床制订鼻咽癌时辰化疗方案提供必要的参数。方法:BALB/C裸小鼠15只,置于程控的独立光照系统中(12h光照,12h黑暗)中同步化至少3周。然后双侧腋下接种人鼻咽低分化鳞癌细胞裸小鼠移植瘤CNE2,成瘤后按灯亮后3、9、15、21h(即3、9、15、21HALO)的时间点取瘤块,制成单个细胞悬液固定,染色后流式细胞仪测DNA含量。用SPSS软件系统ANOVA法检验各期细胞在4个时间点差异的显著性,用Cosinor法考察G1、S、G2/M期细胞在24h的分布是否符合余弦函数。结果:G1、S、G2/M期细胞在3、9、15、21HALO的分布随时间变化有显著性差异,G1、G2/M期细胞变化符合余弦节律。结论:移植于裸鼠的人鼻咽癌细胞的DNA合成随昼夜交替呈节律性变化。

肝脏移植治疗门脉高压症

肝脏移植对门脉高压及其基础肝病是很有效的,许多医生甚至把肝移植视为恶性化进展肝病和门脉高压的首选疗法,因移植可长久地消除门脉高压并发症。但由于器官来源不足,慢性肝病导致门脉高压的患者数远远超过可获得的器官。而且从5年存活率看。

Emerging concepts in liver graft preservation

The urgent need to expand the donor pool in order to attend to the growing demand for liver transplantation has obliged physicians to consider the use of suboptimal liver grafts and also to redefine the preservation strategies. This review examines the different methods of liver graft preservation, focusing on the latest advances in both static cold storage and machine perfusion(MP). The new strategies for static cold storage are mainly designed to increase the fatty liver graft preservation via the supplementation of commercial organ preservation solutions with additives. In this paper we stress the importance of carrying out effective graft washout after static cold preservation, and present a detailed discussion of the future perspectives for dynamic graft preservation using MP at different temperatures(hypothermia at 4 ℃, normothermia a t 3 7 ℃ and subnormothermia at 20 ℃- 2 5 ℃). Finally, we highlight some emerging applications of regenerative medicine in liver graft preservation. In conclusion, this review discusses the "state of the art" and future perspectives in static and dynamic liver graft preservation in order to improve graft viability.

Polyethylene glycol rinse solution:An effective way to prevent ischemia-reperfusion injury

AIM:To test whether a new rinse solution containing polyethylene glycol 35(PEG-35)could prevent ischemia-reperfusion injury(IRI)in liver grafts.METHODS:Sprague-Dawley rat livers were stored in University of Wisconsin preservation solution and then washed with different rinse solutions(Ringer’s lactate solution and a new rinse solution enriched with PEG-35 at either 1 or 5 g/L)before ex vivo perfusion with Krebs-Heinseleit buffer solution.We assessed the following:liver injury(transaminase levels),mitochondrial damage(glutamate dehydrogenase activity),liver function(bile output and vascular resistance),oxidative stress(malondialdehyde),nitric oxide,liver autophagy(Beclin-1 and LCB3)and cytoskeleton integrity(filament and globular actin fraction);as well as levels of metalloproteinases(MMP2 and MMP9),adenosine monophosphate-activated protein kinase(AMPK),heat shock protein 70(HSP70)and heme oxygenase 1(HO-1).RESULTS:When we used the PEG-35 rinse solution,reduced hepatic injury and improved liver function were noted after reperfusion.The PEG-35 rinse solution prevented oxidative stress,mitochondrial damage,and liver autophagy.Further,it increased the expression of cytoprotective heat shock proteins such as HO-1 and HSP70,activated AMPK,and contributed to the restoration of cytoskeleton integrity after IRI.CONCLUSION:Using the rinse solution containing PEG-35 was effective for decreasing liver graft vulnerability to IRI.

Differences and similarities between mesenchymal stem cell and endothelial progenitor cell immunoregulatory properties against T cells

Bone-marrow-derived mesenchymal stem cells and endothelial progenitor cells have some interesting biological properties that make them unique for cell therapy of degenerative and cardiovascular disorders.Although both cell populations have been already studied and used for their regenerative potentials,recently their special immunoregulatory features have brought much more attention.Mesenchymal stem cells and endothelial progenitor cells have both proangiogenic functions and have been shown to suppress the immune response,particularly T cell proliferation,activation,and cytokine production.This makes them suitable choices for allogeneic stem cell transplantation.Nevertheless,these two cells do not have equal immunoregulatory activities.Many elements including their extraction sources,age/passage,expression of different markers,secretion of bioactive mediators,and some others could change the efficiency of their immunosuppressive function.However,to our knowledge,no publication has yet compared mesenchymal stem cells and endothelial progenitor cells for their immunological interaction with T cells.This review aims to specifically compare the immunoregulatory effect of these two populations including their T cell suppression,deactivation,cytokine production,and regulatory T cells induction capacities.Moreover,it evaluates the implications of the tumor necrosis factor alpha-tumor necrosis factor receptor 2 axis as an emerging immune checkpoint signaling pathway controlling most of their immunological properties.

Genomic integrity of human induced pluripotent stem cells:Reprogramming, differentiation and applications

Ten years after the initial generation of induced pluripotent stem cells(hiPSCs)from human tissues,their potential is no longer questioned,with over 15000 publications listed on PubMed,covering various fields of research;including disease modeling,cell therapy strategies,pharmacology/toxicology screening and 3D organoid systems.However,despite evidences that the presence of mutations in hiPSCs should be a concern,publications addressing genomic integrity of these cells represent less than 1%of the literature.After a first overview of the mutation types currently reported in hiPSCs,including karyotype abnormalities,copy number variations,single point mutation as well as uniparental disomy,this review will discuss the impact of reprogramming parameters such as starting cell type and reprogramming method on the maintenance of the cellular genomic integrity.Then,a specific focus will be placed on culture conditions and subsequent differentiation protocols and how their may also trigger genomic aberrations within the cell population of interest.Finally,in a last section,the impact of genomic alterations on the possible usages of hiPSCs and their derivatives will also be exemplified and discussed.We will also discuss which techniques or combination of techniques should be used to screen for genomic abnormalities with a particular focus on the necessary quality controls and the potential alternatives.

Pathological evolution of hepatitis C virus-“Healthy carriers”

AIM: To determine factors associated with fibrosis progression in hepatitis C virus (HCV)-infected patients without signifi cant initial pathological lesions. METHODS: Seventy six untreated HCV-infected pa-tients with initially normal liver as defi ned by a Knodell score≤3,with 2 liver biopsies and detectable HCV-RNA were included. Markers of fibrosis progression were assessed. RESULTS: Median duration of infection and time between paired biopsies was 13 (95% CI: 1-28) and 4 (95% CI: 2-16) years respectively. Alanine-transaminase (ALT) activity was normal in 43.4% of cases. 50% demonstrated progression of the necro-in? ammation and 34% of fi brosis after a median time evolution of 4 years (95% CI: 2-16). The median dif-ference in the necro-inflammation and fibrosis score between biopsies was low,1.5 and 0.0 respectively. Univariate analysis showed there was no difference between fibrosis activity or evolution according to genotype or viral load. A higher fibrosis progression (P = 0.03) was observed in patients with body mass index (BMI) > 25. Fibrosis progression correlated with the time interval between biopsies (P = 0.01). A sig-nifi cant progression of activity (1.7 vs 0.4,P < 0.05) or fi brosis (0.9 vs 0.0,P < 0.01) was observed in patients with elevated ALT. There was a signifi cant correlation between activity progression and fi brosis progression (P = 0.003). Multivariate analysis demonstrated that fi brosis progression was associated with elevated ALT,BMI > 25 and the time interval between 2 biopsies. CONCLUSION: There is no fibrosis progression in 66% of patients without signifi cant initial histopatho-logical lesion. Fibrosis progression is associated with elevated ALT and BMI > 25.

Solitary pulmonary metastasis arising thirteen years after liver transplantation for HBV-related hepatocellular carcinoma

我们描述了为复杂化肝炎 B (HBV ) 的肝细胞癌(HCC ) 在 1989 经历了肝移植的一个 59 岁的男病人肝硬化。在 2001 (在肝移植以后的 12 年) ，没有肝内复发，他开发了 HCC 的肺转移，切除术被做。在 2003 年 7 月，他是没有任何复发免费的症状。HCC 转移罐头甚至在肝移植的一很长的时间以后发展。许多 HCC 慢慢地成长，并且在收到抑制免疫力的治疗的病人的再发性瘤的生长率比不收到抑制免疫力的治疗的那些的显著地大。

Spinal cord injury reprograms muscle fibroadipogenic progenitors to form heterotopic bones within muscles

The cells of origin of neurogenic heterotopic ossifications(NHOs), which develop frequently in the periarticular muscles following spinal cord injuries(SCIs) and traumatic brain injuries, remain unclear because skeletal muscle harbors two progenitor cell populations: satellite cells(SCs), which are myogenic, and fibroadipogenic progenitors(FAPs), which are mesenchymal. Lineage-tracing experiments using the Cre recombinase/Lox P system were performed in two mouse strains with the fluorescent protein Zs Green specifically expressed in either SCs or FAPs in skeletal muscles under the control of the Pax7 or Prrx1 gene promoter, respectively. These experiments demonstrate that following muscle injury, SCI causes the upregulation of PDGFRα expression on FAPs but not SCs and the failure of SCs to regenerate myofibers in the injured muscle, with reduced apoptosis and continued proliferation of muscle resident FAPs enabling their osteogenic differentiation into NHOs. No cells expressing Zs Green under the Prrx1 promoter were detected in the blood after injury, suggesting that the cells of origin of NHOs are locally derived from the injured muscle. We validated these findings using human NHO biopsies. PDGFRα+mesenchymal cells isolated from the muscle surrounding NHO biopsies could develop ectopic human bones when transplanted into immunocompromised mice, whereas CD56+myogenic cells had a much lower potential. Therefore, NHO is a pathology of the injured muscle in which SCI reprograms FAPs to undergo uncontrolled proliferation and differentiation into osteoblasts.

Conversion of Unresectable to Resectable Liver Metastases from Colorectal Carcinoma Using Hepatic Arterial Chronomodulated Chemotherapy: A Case Report and Short Literature Review

Background: The regional chronomodulated hepatic arterial infusion chemotherapy (HAIC) is an effective regimen for the treatment of patients with unresectable liver metastases from colorectal cancer, especially for the conversion into resectability. Aim: To demonstrate that chronomodulated HAI triplet chemotherapy according to OPTILIV protocol is well tolerated and displayed high antitumor activity in this heavily-pretreated patient. Case Presentation: A 54 years old patient from Russia was treated for a tumor in the ascending colon presented with 13 hepatic metastases ranging from 0.3 to 2.7 cm in diameter. He underwent a laparoscopic right hemicolectomy, 12 cycles of FOLFIRINOX combined to bevacizumab for the last 5 cycles, resulting in a partial response according to CT scan. It was decided to perform a two-stage hepatectomy at Paul Brousse hospital: left partial hepatectomy allowed the excision of 9 lesions. Radio frequency ablation was performed in 2 nodular lesions. Afterwards, the patient received 5 cycles of chronomodulated triplet chemotherapy into the hepatic artery, according to the OPTILIV protocol design, yet without cetuximab, because of the KRAS mutation in the liver metastases, with a partial re-sponse. The patient could then undergo the second stage of the planned right hepatectomy, which turned out to be an R0 resection followed by receiving three courses of chronomodulated HAIC. Disease progression was documented after 3 months. Chronomodulated FOLFIRI chemotherapy was re-started intravenously, in combination with Aflibercept and it was associated with further disease progression. The genetic analysis of our patient’s cancer revealed a high level of MSI. The patient was included in the Phase 2 CheckMate-142 trial and received nivolumab 3 mg/kg every 2 weeks within 3 months. Treatment was discontinued due to ineffectiveness. Then the patient underwent radiotherapy geared towards reduction of pain. Afterwards, the patient died from the disease progression 2 years after the beginning of treatment. Conclusion: In this article, the authors report a clinical case with chronomodulated HAIC as rescue therapy in a heavily pretreated patient with metastatic colorectal cancer, allowing to achieve an objective response despite prior progression on FOLFIRINOX (the same triplet chemo by IV route). This strategy permitted to overcome drug resistance and to perform further complete resection of the liver me-tastases with prolonged patient survival. Thus, chronomodulated HAI is useful in patients with liver metastases from colorectal cancer and de-serves to be further assessed prospectively in clinical trials chemotherapy.

应用特异性IgG亲和力诊断甲型肝炎

Aim -Diagnosis of acute hepatitis A virus (HAV) infection is classically based on the detection of HAV-IgM. Nevertheless, HAV-IgM can be positive for patients with polyclonal stimulation of their immune system (i.e. immune reactivation). To improve the diagnostic yield, an avidity test for HAV-IgG antibodies was developed and tested. Methods -Avidity tests were performed in 128 sera: 11 selected samples from patients with past infection, 15 acute hepatitis A, 10 vaccinated subjects and 4 patients with immune reactivation as well as 84 HAV IgM positive unselected sera, provided by routine laboratories. Results -Patients with past infection had avidities over 70%, whereas avidities in patients with acute hepatitis A were below 50%during the first month following the onset of symptoms. As expected, patients with immune reactivation had avidities over 70%consistent with past infection. The results obtained for the 84 unselected sera allowed reconsidering the diagnosis of acute hepatitis A for nearly a third of patients. Conclusion -This test could improve the diagnosis of acute hepatitis A infection, particularly in elderly patients.

熊脱氧胆酸缓解硫唑嘌呤和皮质激素耐药的I型自身免疫性肝炎

Combination therapy with steroids and azathioprine is the reference treatment for autoimmune hepatitis, but potential adverse effects are numerous and intolerance can occur. We report a patient with a well-documented type 1 autoimmune hepatitis intolerant to corticosteroids and azathioprine therapy, in whom eight years of ursodeoxycholic acid monotherapy was associated with biochemical and histological remission.

胎儿至成年期女性肝脏中的男性细胞微嵌合现象

Male microchimerismis frequent in the adult female liver and is attributed to fetal cells originating frompreviousmale offspring. It has never been studied in pregnant women, female children, or fetuses. We examined its frequency and cellular nature in normal and diseased female livers from fetal life to adulthood. Forty-six liver samples from 29 women, 6 female children, and 11 female fetuses were screened for the Y chromosome via polymerase chain reaction (PCR) assay and fluorescent in situ hybridization (FISH). The X chromosome was used as an internal control. A third PCR assay was used for Y genotyping. The Y chromosome was detected in 5 of 6 children, 7 of 11 fetuses, 3 of 9 women with normal liver, 7 of 10 women with chronic hepatitis C, 5 of 6 women with acute liver disease during pregnancy with male offspring, and 2 of 4 nonpregnant women with fulminant hepatitis. In positive samples, the mean XY/XX ratio was 0.012 (±0.004). In women, male microchimerism was correlated with previous male offspring. Male hepatocytes, detected via FISH combined with anti-hepatocyte immunohistochemistry,were observed only in fetuses (4/9) and in postpartem women (4/6). Y genotypes were different from each other in 4 of 5 female livers. In conclusion, male liver microchimerism is frequent in normal and diseased female livers. The presence of male cells in the liver of female children and fetuses is probably due to the transplacental transmission of fetal cells preexisting in the mother and acquired either from previous pregnancy with male offspring or during the mother’s own fetal life.

Quality of life 10 years after liver transplantation:The impact of graft histology

AIM To evaluate the relationship between the state of transplanted liver graft and the recipient quality of life(QOL) of histologically proven lesions in a 10-year post liver transplantation(LT) cohort of patients. METHODS Seventy-two recipients with a functional first graft at 10 years post-LT underwent liver biopsy and completed a QOL questionnaire. Logistic regression analysis was used to explore associations between histological, clinical andQOL criteria. RESULTS Ten years after LT, fibrosis was detected in 53% of patients, and affected the general health perception, while ductopenia, present in 36%, affected the well-being(P = 0.05). Hepatic steatosis(HS) was present in 33% of patients and was associated with the worst QOL score on multiple domains. When compared to patients without HS, patients with HS had significantly higher incidence of fibrosis(P = 0.03), hepatitis C virus(HCV) infection(P = 0.007), and more patients had retired from their job(P = 0.03). Recurrent or de novo HCV-associated fibrosis and patient retirement as objective variables, and abdominal pain or discomfort and joint aches or pains as subjective variables, emerged as independent determinants of HS. CONCLUSION Long-term liver graft lesions, mainly HS presumably as a surrogate marker of HCV infection, may have a substantial impact on QOL 10 years after LT.

Impact of body mass index on hepatocellular carcinoma recurrence after liver transplantation through long-term follow-up

Background:Obesity is associated with increased oncological risk and outcomes but the evidence surrounding the effect of body mass index(BMI)on increased risk of hepatocellular carcinoma(HCC)recurrence after liver transplantation(LT)is still questionable.The purpose of this retrospective study of a large cohort of adult patients transplanted for HCC was to investigate the effect of BMI on the incidence of HCC recurrence and outcome.Methods:Data from 427 adult recipients transplanted for HCC between 2000 and 2017 were collected.Patients were classified at time of LT according to the World Health Organization BMI classification into 3 groups;group 1:BMI<25(n=166),group 2:BMI 25-29.9(n=150)and group 3:BMI≥30(n=111).Results:There were no significant changes of mean BMI overtime 26.8±5.0 kg/m2 at time of LT and 28.8±23.1 at 5 years.The recurrence rates of HCC after LT in the three groups were 19%,16%and 17%respectively.The 5,10 and 15-year recurrence free survival(RFS)rates were respectively 68.6%,47.3%and 40.8%in group 1,73.3%,66.2%and 49.5%in group 2 and 68.8%,57.5%and 47.7%in group 3(log rank P=0.47).Conclusions:Recipient BMI at time of transplant and during follow-up didn’t impact the incidence of HCC recurrence nor long-term patient survival,irrespective to the status of the patients and their tumor characteristic at time of LT.The present study clearly confirms that obesity should not be considered,when selecting patients with HCC to LT,as a predictive factor of recurrence.

Patient with unresectable colorectal liver metastases and asymptomatic primary tumor:end of the debate!

In patients with definitively unresectable colorectal liver metastases,whether the primary tumor should be resected prior to chemotherapy is an old question.Several retrospective studies(1-5)and a meta-analysis with individual data(6)had suggested that primary resection of the primary tumor followed by chemotherapy was associated with better survival than the chemotherapy-first strategy.The prevention of complications related to the primary tumor(occlusion,perforation,bleeding)as well as a better response to chemotherapy were usual justifications to explain the superiority of primary tumor resection-first.However,the absence of any randomized study made it difficult to conclude on this question due to the numerous selection biases related to these retrospective analyses.

The quest for optimal and reliable guidelines based on robust evidence for the treatment of cholangiocarcinoma

In 1991,Gordon H.Guyatt described the evidence-based medicine(EBM)as“a focus on educating front-line clinicians in assessing the credibility of research evidence,understanding the results of clinical studies,and determining how best to apply the results to their everyday practice”(1).In 2010,Graham et al.defined clinical guidelines as“statements that include recommendations intended to optimize patient care that are informed by a systematic review of the evidence and an assessment of the benefits and harms of alternative care options”(2).Since 2010,the number of clinical guidelines increased exponentially,and the query on PubMed with the term“new guidelines”produces 59,773 results.