Overexpression of wild-type HRAS drives non-alcoholic steatohepatitis to hepatocellular carcinoma in mice

Hepatocellular carcinoma(HCC),a prevalent solid carcinoma of significant concern,is an aggressive and often fatal disease with increasing global incidence rates and poor therapeutic outcomes.The etiology and pathological progression of non-alcoholic steatohepatitis(NASH)-related HCC is multifactorial and multistage.However,no single animal model can accurately mimic the full NASH-related HCC pathological progression,posing considerable challenges to transition and mechanistic studies.Herein,a novel conditional inducible wild-type human HRAS overexpressed mouse model(HRAS-HCC)was established,demonstrating 100%morbidity and mortality within approximately one month under normal dietary and lifestyle conditions.Advanced symptoms of HCC such as ascites,thrombus,internal hemorrhage,jaundice,and lung metastasis were successfully replicated in mice.In-depth pathological features of NASH-related HCC were demonstrated by pathological staining,biochemical analyses,and typical marker gene detections.Combined murine anti-PD-1 and sorafenib treatment effectively prolonged mouse survival,further confirming the accuracy and reliability of the model.Based on protein-protein interaction(PPI)network and RNA sequencing analyses,we speculated that overexpression of HRAS may initiate the THBS1-COL4A3 axis to induce NASH with severe fibrosis,with subsequent progression to HCC.Collectively,our study successfully duplicated natural sequential progression in a single murine model over a very short period,providing an accurate and reliable preclinical tool for therapeutic evaluations targeting the NASH to HCC continuum.

Update on biomarkers of glycemic control

Attaining and maintaining good glycemic control is a cornerstone of diabetes care. The monitoring of glycemic control is currently based on the self-monitoring of blood glucose(SMBG) and laboratory testing for hemoglobin A1 c(HbA1 c),which is a surrogate biochemical marker of the average glycemia level over the previous 2-3 mo period. Although hyperglycemia is a key biochemical feature of diabetes, both the level of and exposure to high glucose, as well as glycemic variability, contribute to the pathogenesis of diabetic complications and follow different patterns in type 1 and type 2 diabetes. HbA1 c provides a valuable,standardized and evidence-based parameter that is relevant for clinical decision making, but several biological and analytical confounders limit its accuracy in reflecting true glycemia. It has become apparent in recent years that other glycated proteins such as fructosamine, glycated albumin, and the nutritional monosaccharide 1,5-anhydroglucitol, as well as integrated measures from direct glucose testing by an SMBG/continuous glucose monitoring system, may provide valuable complementary data, particularly in circumstances when HbA1 c results may be unreliable or are insufficient to assess the risk of adverse outcomes. Long-term associations of these alternative biomarkers of glycemia with the risk of complications need to be investigated in order to provide clinically relevant cut-off values and to validate their utility in diverse populations of diabetes patients.

Transfusion-transmitted virus in association with hepatitis A-E viral infections in various forms of liver diseases in India

AIM： To describe the prevalence of transfusiontransmitted virus （TTV） infection in association with hepatitis A-E viral infections in different forms of liver diseases in North India. METHODS： Sera from a total number of 137 patients, including 37 patients with acute viral hepatitis （AVH）, 37 patients with chronic viral hepatitis （CVH）, 31 patients with cirrhosis of liver and 32 patients with fulminant hepatic failure （FHF）, were analyzed both for TTV-DNA and hepatitis A-E viral markers. Presence of hepatitis B virus （HBV）, hepatitis C virus （HCV） and hepatitis E virus （HEV） infections was detected in different proportions in different groups. Moreover, rrV-DNA was simultaneously tested in 100 healthy blood donors also. RESULTS： None of the patients had hepatitis A virus （HAV） and hepatitis D virus （HDV） infections. Overall prevalence of TTV-DNA was detected in 27.1% cases with AVH, 18.9% cases with CVH, 48.4% cases with cirrhosis and 9.4% cases with FHF. TTV-DNA simultaneously tested in 100 healthy blood donors showed 27% positivity. On establishing a relation between TTV infection with other hepatitis viral infections, TTV demonstrated co-infection with HBV, HCV and HEV in these disease groups. Correlation of TTV with ALT level in sera did not demonstrate high ALT level in TTV-infected patients, suggesting that TI-V does not cause severe liver damage. CONCLUSION： TTV infection is prevalent both in patients and healthy individuals in India. However, it does not have any significant correlation with other hepatitis viral infections, nor does it produce an evidence of severe liver damage in patients with liver diseases.

Left atrial thrombosis in an anticoagulated patient after bioprosthetic valve replacement:Report of a case

We present the case of a 74 year old woman suffering from severe mitral valve incompetence and rapid atrial fibrillation. After an appropriate vitamin K antagonist(VKA) therapy, the patient underwent mitral valve replacement by bioprosthesis. Then, the patient was rehospitalized for jaundice. Suspecting hepatotoxicity, VKA was discontinued and fondaparinux was started. During this treatment, the patient developed a symptomatic atrial thrombus. After exclusion of a hepatic disease, VKA was re-established with hemodynamic and liver enzymes normalization and atrial thrombus resolution. Caution has to be used when considering fondaparinux as an alternative strategy to VKA in patients with multiple thrombotic risk factors.

Alternative splicing of DNA damage response genes and gastrointestinal cancers

Alternative splicing,which is a common phenomenon in mammalian genomes,is a fundamental process of gene regulation and contributes to great protein diversity.Alternative splicing events not only occur in the normal gene regulation process but are also closely related to certain diseases including cancer.In this review,we briefly demonstrate the concept of alternative splicing and DNA damage and describe the association of alternative splicing and cancer pathogenesis,focusing on the potential relationship of alternative splicing,DNA damage,and gastrointestinal cancers.We will also discuss whether alternative splicing leads to genetic instability,which is considered to be a driving force for tumorigenesis.Better understanding of the role and mechanism of alternative splicing in tumorigenesis may provide new directions for future cancer studies.

Combined Effects of Blood Pressure and Aldosterone on Cardiac Left Ventricular Mass Index—Ethnic Differences between Kazakh, Uygur and Han Subjects

Previous Background: Hemodynamic factors, like blood pressure, have been established to be major determinants of cardiac left ventricular structure. However, several factors other than blood pressure to influence cardiac mass have been implicated. When we did medical survey, cardiac left ventricular mass index (LVMI) of one ethnic group that had higher blood pressure was found to be smaller than that of the other ethnic groups with a lower blood pressure. Such contradicted data from the present study were analyzed combining blood pressure, LVMI and chemical parameters obtained from blood and urine. Methods: In a medical survey conducted in Xinjiang, China, 279 people (65 - 70 years old) from three ethnic groups (Kazakh, Uygur and Han) from two separated regions provided blood and urine samples and underwent echocardiography and 24-h ambulatory blood pressure monitoring (ABPM). Results: Systolic and diastolic blood pressure obtained from ABPM and urinary sodium excretion values were significantly higher in Kazakh than that in Uygur and Han. However, LVMI in Kazakh was lower than that in other 2 groups. Plasma aldosterone concentration (PAC) and plasma renin activity (PRA) were significantly lowest in Kazakh. The values of LVMI in all ethnic groups were positively related to both blood pressure and PAC. An inverse correlation was identified between PAC and urinary sodium excretion value. Conclusion: Although higher blood pressure in Kazakh subjects, their LVMI was lower than those of Uygur and Han, whose blood pressure was lower than that in Kazakh. These results suggest that blood pressure is not always a determinant for LVMI value. There is a possibility that relatively lower PAC resulted from higher sodium intake suppressed the rise in LVMI caused by higher blood pressure in Kazakh.

One-Step Multiplex PCR for Simultaneous Detection and Identification of Eight Medically Important <i>Candida</i>Species

Recently, the incidence of Candida infections has substantially increased. Conventional identification methods for Candida species are technically difficult to conduct and cannot accurately distinguish each species. The purpose of the present study was to design primers to identify and detect simultaneously eight medically important Candida species using one-step multiplex PCR. PCR primers were designed based on partial sequences of intergenic spacer (IGS) and internal transcribed spacer (ITS) genes of eight medically important Candida species. These primers were able to distinguish each Candida species and did not display cross-reactivity with representative Candida species other than the eight Candida species. Moreover, our developed one-step multiplex PCR method is accurate, specific, cost-effective, time-saving, and worked without requiring DNA extraction.

Vibration in mice: A review of comparative effects and use in translational research

Sound pressure waves surround individuals in everyday life and are perceived by animals and humans primarily through sound or vibration. When sound pressure waves traverse through a solid medium, vibration will result. Vibration has long been considered an unwanted variable in animal research and may confound scientific endeavors using animals. Understanding the characteristics of vibration is required to determine whether effects in animals are likely to be therapeutic or result in adverse biological effects. The eighth edition of the "Guide for the Care and Use of Laboratory Animals" highlights the importance of considering vibration and its effects on animals in the research setting, but knowledge of the level of vibration for eliciting these effects was unknown. The literature provides information regarding therapeutic use of vibration in humans, but the range of conditions to be of therapeutic benefit is varied and without clarity. Understanding the characteristics of vibration(eg, frequency and magnitude) necessary to cause various effects will ultimately assist in the evaluation of this environmental factor and its role on a number of potential therapeutic regimens for use in humans. This paper will review the principles of vibration, sources within a research setting, comparative physiological effects in various species, and the relative potential use of vibration in the mouse as a translational research model.

A New Automated Method and Sample Data Flow for Analysis of Volatile Nitrosamines in Human Urine

Volatile nitrosamines (VNAs) are a group of compounds classified as probable (group 2A) and possible (group 2B) carcinogens in humans. Along with certain foods and contaminated drinking water, VNAs are detected at high levels in tobacco products and in both mainstream and side-stream smoke. Our laboratory monitors six urinary VNAs—N-nitrosodimethylamine (NDMA), N-nitrosomethylethylamine (NMEA), N-nitrosodiethylamine (NDEA), N-nitrosopiperidine (NPIP), N-nitrosopyrrolidine (NPYR), and N-nitrosomorpholine (NMOR)—using isotope dilution GC-MS/ MS (QQQ) for large population studies such as the National Health and Nutrition Examination Survey (NHANES). In this paper, we report for the first time a new automated sample preparation method to more efficiently quantitate these VNAs. Automation is done using Hamilton STARTM and Caliper StaccatoTM workstations. This new automated method reduces sample preparation time from 4 hours to 2.5 hours while maintaining precision (inter-run CV < 10%) and accuracy (85% - 111%). More importantly this method increases sample throughput while maintaining a low limit of detection (<10 pg/mL) for all analytes. A streamlined sample data flow was created in parallel to the automated method, in which samples can be tracked from receiving to final LIMs output with minimal human intervention, further minimizing human error in the sample preparation process. This new automated method and the sample data flow are currently applied in bio-monitoring of VNAs in the US non-institutionalized population NHANES 2013-2014 cycle.

Cohort analysis of pediatric intussusception score to diagnose intussusception

BACKGROUND Intussusception is a primary cause of intestinal obstruction in young children.Delayed diagnosis is associated with increased morbidity.Ultrasonography(USG)is the gold standard for diagnosis,but it is operator dependent and often unavailable in limited resource areas.AIM To study the clinical characteristics of intussusception including management and evaluation of the diagnostic accuracy of abdominal radiography(AR)and the promising parameters found in the pediatric intussusception score(PIS).METHODS Children with suspected intussusception in our center from 2006 to 2018 were recruited.Clinical manifestations,investigations,and treatment outcomes were recorded.AR images were interpreted by a pediatric radiologist.Diagnosis of intussusception was composed of compatible USG and response with reduction.The diagnostic value of the proposed PIS was evaluated.RESULTS Ninety-seven children were diagnosed with intussusception(2.06±2.67 years,62.9%male),of whom 74%were<2 years old and 37.1%were referrals.The common manifestations of intussusception were irritability or abdominal pain(86.7%)and vomiting(59.2%).Children aged 6 mo to 2 years,pallor,palpable abdominal mass,and positive AR were the parameters that could discriminate intussusception from other mimics(P<0.05).Referral case was the only significant parameter for failure to reduce intussusception(P<0.05).AR to diagnose intussusception had a sensitivity of 59.2%.The proposed PIS,a combination of clinical irritability or abdominal pain,children aged 6 mo to 2 years,and compatible AR,had a sensitivity of 85.7%.CONCLUSION AR alone provides poor screening for intussusception.The proposed PIS in combination with common manifestations and AR data was shown to increase the diagnostic sensitivity,leading to timely clinical management.

Simultaneous Occurrence of Myofascial Pain Referred to Tooth and Endodontic Lesions in Patient with Toothache: A Case Report

Toothache is almost always caused by odontogenic toothache, but diagnosis is more difficult in the case of nonodontogenic toothache. We report a case of simultaneous occurrence of odontogenic and nonodontogenic toothache. This manuscript presents a case report for a 35-year-old woman who visited our Orofacial and Head Pain Clinic with the chief complaint of continuous dull pain in left maxillary molar teeth region. It was concluded to be a case of simultaneous odontogenic toothache and nonodontogenic toothache. It was successfully treated by an endodontist and an orofacial pain specialist. The endodontist performed root canal treatment against odontogenic toothache caused by apical periodontitis using a dental operating microscope. The presence of a trigger point (TP) resulting in tooth pain was inferred. A trigger point injection (TPI) was administered by orofacial pain specialist, and toothache relief was confirmed. Myofascial pain was diagnosed definitively. After confirming that the toothache had resolved at multiple TPIs, a crown prosthesis was placed. Following the application of crown prosthesis, we were concerned but did not find recurrence of toothache from myofascial pain due to increased occlusal force. This case suggests that there is no single cause of chronic pain and that multiple causes must be considered for diagnosis, suggesting the need for treatment by multiple specialists.

Reference values for selected hematological, biochemical and physiological parameters of Sprague‐Dawley rats at the Animal House, Faculty of Medicine, University of Colombo,Sri Lanka

Background: Lack of available reference values in a research setting under local conditions can be a drawback for beginners, as the accuracy of data from control samples cannot be checked at the beginning of a research project. This affects comparisons with data from test samples. To avoid these complications in their research projects,beginners tend to have a greater number of animals in the control group compared to test groups in order to have control group measurements within 2 SDs of the mean.Methods: As non-availability of reference values was a long-felt need, the described project was conducted in order to establish a reference database for selected haematological, biochemical and physiological parameters using apparently healthy Sprague-Dawley rats bred in the Animal House of Faculty of Medicine, University of Colombo(UCFM).Results: Differences in mean values of packed cell volume(PCV), mean corpuscular volume(MCV) and mean corpuscular hemoglobin concentration(MCHC), serum creatinine and blood glucose levels between the two genders were statistically significant. Lipid profile measurements did not differ significantly between genders, but mean and median values of triglycerides(TG) between male and female rats showed a difference of more than 10 mg/dL. The liver enzymes alkaline phosphatase(AP)and aspartate aminotransferase(AST) were also statistically significantly different between sexes. Despite wide variation in mean alanine aminotransferase(ALT)between sexes, the difference was not statistically significant.Conclusion: The findings of this project should support to a certain extent the"Reduction" aspect of the 3 Rs concept of Russell and Burch by reducing the number of Sprague-Dawley rats used in future research projects at UCFM.

The Transcriptional Activity of LAMP3 Gene Involved in Autophagocytosis in Colorectal Cancer LAMP3 Expression in Colorectal Cancer

Background: Autophagocytosis is a biological process involving the controlled distribution of cell fragments and organelles in order to obtain an additional source of energy. LAMP3 (lysosome-associated membrane protein 3) is a heavily glycosylated integral membrane protein located mainly in the lysosomal membrane. Recent studies have shown that it participates in tumor metastasis and drug resistance. Its main role is contribution to tumor cells proliferation, migration and invasion. The aim of this study was to determine changes in transcriptional activity of LAMP3 at various stages of colorectal cancer. In addition, an attempt was made to select miRNAs potentially regulating LAMP3 expression using bioinformatic databases. Methods: The study was conducted on healthy colon samples and colon cancer samples in four stages. Molecular analysis included the extraction of total RNA, purification of the obtained extracts, expression profile analysis using oligonucleotide microarray technique and in silico determination of miRNAs potentially regulating the LAMP3 expression. Results: The level of LAMP3 expression is higher in colorectal adenocarcinoma cells than in non-cancerous cells and depends on the stage of the disease. Conclusions: LAMP3 may promote cancer progression, metastasis and cause the resistance to treatment.

circRNA3669 promotes goat endometrial epithelial cells proliferation via miR-26a/RCN2 to activate PI3K/AKT-mTOR and MAPK pathways

The development of receptive endometrium(RE) from pre-receptive endometrium(PE) for successful embryo implantation is a complex dynamic process in which the morphology and physiological states of the endometrial epithelium undergo a series of significant changes, including cell proliferation and apoptosis. However, the molecular mechanisms are not yet fully understood. In this study, a higher circRNA3669 level was observed in PE than in RE of goats. Functional assays revealed that this overexpression promoted the proliferation of goat endometrial epithelial cells(GEECs) by activating the expression of genes related to the PI3K/AKT-mTOR and MAPK pathways,thereby inhibiting apoptosis in vitro. Furthermore, circRNA3669 functioned as a competing endogenous RNA(ceRNA) to upregulate Reticulocalbin-2(RCN2) expression at the post-transcriptional level by interacting with and downregulating miR-26a in GEECs. In addition, RCN2, which is highly expressed in the PE of goats, was found to be regulated by β-estradiol(E2) and progesterone(P4). Our results demonstrated that RCN2 also affected the key proteins PI3K, AKT, mTOR, JNK, and P38 in the PI3K/AKT-mTOR and MAPK pathways, thereby facilitating GEECs proliferation and suppressing their apoptosis in vitro. Collectively, we constructed a new circRNA3669-miR-26aRCN2 regulatory network in GEECs, which further provides strong evidence that circRNA could potentially play a crucial regulatory role in the development of RE in goats.

Resolving tumor evolution:a phylogenetic approach

The evolutionary dynamics of cancer,characterized by its profound heterogeneity,demand sophisticated tools for a holistic understanding.This review delves into tumor phylogenetics,an essential approach bridging evolu-tionary biology with oncology,offering unparalleled insights into cancer’s evolutionary trajectory.We provide an overview of the workflow,encompassing study design,data acquisition,and phylogeny reconstruction.No-tably,the integration of diverse data sets emerges as a transformative step,enhancing the depth and breadth of evolutionary insights.With this integrated perspective,tumor phylogenetics stands poised to redefine our understanding of cancer evolution and influence therapeutic strategies.

基于直向同源序列的比较基因组学研究

直向同源序列在不同的物种中具有相近甚至相同的功能、相似的调控途径,扮演相似甚至相同的角色,而且,绝大多数核心生物功能就是由相当数量的直向同源基因所承担,它是基因组序列的功能注释与分析中最可靠的选择,其特殊的生物学特性决定:利用直向同源序列开展比较基因组学研究,必将为探测不同生物在进化过程中重要功能基因的出现、表达和丢失提供线索。文章从直向同源基因的基本特性、直向同源序列与比较基因组学的关系、应用直向同源序列开展比较基因组学相关研究方法、现状等展开综述。

Triglyceride levels and apolipoprotein E polymorphism in patients with acute pancreatitis

BACKGROUND:Hypertriglyceridemia is an unusual cause of acute pancreatitis and sometimes considered to be an epiphenomenon.This study aimed to investigate the clinical and analytical features and the APOE genotypes in patients with acute pancreatitis and severe hypertriglyceridemia.METHODS:We undertook a one-year,prospective study of patients with acute pancreatitis whose first laboratory analysis on admission to the emergency department included measurement of serum triglycerides.The APOE genotype was determined and the patients answered an established questionnaire within the first 24 hours concerning their alcohol consumption,the presence of co-morbidities and any medications being taken.The patients’ progression,etiological diagnosis,hospital stay and clinical and radiological severity were all recorded.RESULTS:Hypertriglyceridemia was responsible for 7 of 133 cases of pancreatitis (5%);the remaining cases were of biliary (53%),idiopathic (26%),alcoholic (11%) or other (5%) origin.Compared with these remaining cases,the patients with hypertriglyceridemia were significantly younger,had more relapses,and more often had diabetes mellitus.They usually consumed alcohol or consumed it excessively on the days before admission.Also,the ε4 allele of the APOE gene was more common in this group (P<0.05).CONCLUSION:One of 20 episodes of acute pancreatitis is caused by hypertriglyceridemia and it is linked to genetic (ε4 allele) and comorbid factors such as diabetes and,especially,alcohol consumption.

Challenges and improvements in testosterone and estradiol testing

Assays that measure steroid hormones in patient care, public health, and research need to be both accurate and precise, as these criteria help to ensure comparability across all clinical and research applications. This review addresses major issues relevant to assay variability and describes recent activities by the US Centers for Disease Control and Prevention （CDC） to improve assay performance. Currently, high degrees of accuracy and precision are not always met for testosterone and estradiol measurements; although technologies for steroid hormone measurement have advanced significantly, measurement variability within and across laboratories has not improved accordingly. Differences in calibration and specificity are discussed as sources of variability in measurement accuracy. Ultimately, a combination of factors appears to cause inaccuracy of steroid hormone measurements, with nonuniform assay calibration and lack of specificity being two major contributors to assay variability. Within-assay variability for current assays is generally high, especially at low analyte concentrations. The CDC Hormone Standardization （HoSt） Program is improving clinical assays, as evidenced by a 50% decline in mean absolute bias between mass spectrometry assays and the CDC reference method from 2007 to 2011. This program provides the measurement traceability to CDC reference methods and helps to minimize factors affecting measurement variability.