The thermoneutral zone in women takes an "arctic" shift compared to men.

Conventionally, women are perceived to feel colder than men, but controlled comparisons are sparse. We measured the response of healthy, lean, young women and men to a range of ambient temperatures typical of the daily environment (17 to 31 °C). The Scholander model of thermoregulation defines the lower critical temperature as threshold of the thermoneutral zone, below which additional heat production is required to defend core body temperature. This parameter can be used to characterize the thermoregulatory phenotypes of endotherms on a spectrum from "arctic" to "tropical." We found that women had a cooler lower critical temperature (mean ± SD: 21.9 ± 1.3 °C vs. 22.9 ± 1.2 °C, P = 0.047), resembling an "arctic" shift compared to men. The more arctic profile of women was predominantly driven by higher insulation associated with more body fat compared to men, countering the lower basal metabolic rate associated with their smaller body size, which typically favors a "tropical" shift. We did not detect sex-based differences in secondary measures of thermoregulation including brown adipose tissue glucose uptake, muscle electrical activity, skin temperatures, cold-induced thermogenesis, or self-reported thermal comfort. In conclusion, the principal contributors to individual differences in human thermoregulation are physical attributes, including body size and composition, which may be partly mediated by sex.

Did implementation of no-fault auto-insurance in British Columbia, Canada, impact return to work following road trauma? Protocol for a before-after survival analysis.

INTRODUCTION: Road trauma (RT) is a major public health problem that often results in prolonged absenteeism from work. Limited evidence suggests that recovery after RT is associated with automobile insurance compensation schemes. In May 2021, British Columbia, Canada switched from fault-based to no-fault auto-insurance coverage. This manuscript presents the protocol for a planned evaluation of that natural experiment: We will evaluate the impact of changing automobile insurance schemes on return to work following RT. METHODS AND ANALYSIS: The evaluation will use a before-after design to analyse auto-insurance claims (1 April 2019 to 30 April 2024) in order to compare recovery of claimants with non-catastrophic injuries who filed claims under the no-fault insurance scheme to that of those who filed claims under the previous system. Claimants will be followed from date of injury until they return to work or have been followed for 6 months (right-censored). We will perform sensitivity analyses to examine the robustness of our findings. First, we will exclude injuries that occurred during the COVID-19 provincial State of Emergency. Second, we will use propensity score methods rather than conventional covariate adjustment to address potential imbalance between characteristics of claimants pre-change and post-change. Finally, as the implementation effect may have a heterogeneous association with time off work, we will use quantile regression with right-censoring at 6 months to model differences in return to work at the 25th, 50th, 75th and 90th percentiles. ETHICS AND DISSEMINATION: The study uses de-identified data and is approved by the University of British Columbia Clinical Research Ethics Board (H20-03644). This research is funded by the Insurance Corporation of British Columbia (ICBC). Findings will be published in the peer-reviewed literature and summarised in a report prepared for ICBC. We anticipate that our findings will inform policy decisions in other jurisdictions considering switching to no-fault auto-insurance schemes.

Pharmacokinetics and Safety of iGlarLixi in Healthy Chinese Participants: Results of a Phase 1 Randomized Study.

INTRODUCTION: The Chinese Diabetes Society recommends basal insulin and glucagon-like peptide-1 receptor agonists as an add-on therapy to first-line oral antihyperglycemic drugs for people with type 2 diabetes (T2D). Fixed-ratio combination of insulin glargine 100 U/ml (iGlar) and lixisenatide (iGlarLixi) is known to improve glycemic control in adults with T2D. However, the pharmacokinetics of iGlarLixi has not been evaluated in Chinese participants. The present study evaluated pharmacokinetics and safety of two iGlarLixi (10 U/10 µg and 30 U/15 µg) doses following single subcutaneous administration in healthy Chinese participants. METHODS: This was a Phase 1, single-center, open-label, parallel-group, randomized study in healthy Chinese adults who were randomized to receive a single dose of iGlarLixi with either 1:1 (10 U/10 µg) or 2:1 (30 U/15 µg) ratio of iGlar and lixisenatide. Primary objectives include assessment of pharmacokinetics of iGlar in iGlarLixi 30 U/15 µg group and the pharmacokinetics of lixisenatide in both the groups (iGlarLixi 10 U/10 µg and iGlarLixi 30 U/15 µg). Safety and tolerability were also assessed. RESULTS: In iGlarLixi 30 U/15 µg group, iGlar concentrations were low and quantifiable in three of ten participants, while its main metabolite (M1) was quantifiable in all participants, reflecting rapid conversion of iGlar to M1. Median INS-tmax was 14.00 h for iGlar and 13.00 h post-dose for M1. Absorption of lixisenatide was similar in both dose groups with median tmax of 3.25 and 2.00 h post-dose in both groups. The exposure increase was dose proportionate with a 1.5-fold increase in the lixisenatide dose. Adverse events observed were consistent with those previously reported with iGlar or lixisenatide. CONCLUSION: iGlarLixi administration resulted in early absorption of both iGlar and lixisenatide with a good tolerability profile in healthy Chinese participants. These results are consistent with the previously published data from other geographic regions. TRIAL REGISTRATION: U1111-1194-9411.

Continuous glucose monitoring in patients with post-bariatric hypoglycaemia reduces hypoglycaemia and glycaemic variability.

AIM: To determine whether continuous glucose monitoring (CGM) can reduce hypoglycaemia in patients with post-bariatric hypoglycaemia (PBH). MATERIALS AND METHODS: In an open-label, nonrandomized, pre-post design with sequential assignment, CGM data were collected in 22 individuals with PBH in two sequential phases: (i) masked (no access to sensor glucose or alarms); and (ii) unmasked (access to sensor glucose and alarms for low or rapidly declining sensor glucose). Twelve participants wore the Dexcom G4 device for a total of 28 days, while 10 wore the Dexcom G6 device for a total of 20 days. RESULTS: Participants with PBH spent a lower percentage of time in hypoglycaemia over 24 hours with unmasked versus masked CGM (<3.3 mM/L, or <60 mg/dL: median [median absolute deviation {MAD}] 0.7 [0.8]% vs. 1.4 [1.7]%, P = 0.03; <3.9 mM/L, or <70 mg/dL: median [MAD] 2.9 [2.5]% vs. 4.7 [4.8]%; P = 0.04), with similar trends overnight. Sensor glucose data from the unmasked phase showed a greater percentage of time spent between 3.9 and 10 mM/L (70-180 mg/dL) (median [MAD] 94.8 [3.9]% vs. 90.8 [5.2]%; P = 0.004) and lower glycaemic variability over 24 hours (median [MAD] mean amplitude of glycaemic excursion 4.1 [0.98] vs. 4.4 [0.99] mM/L; P = 0.04). During the day, participants also spent a greater percentage of time in normoglycaemia with unmasked CGM (median [MAD] 94.2 [4.8]% vs. 90.9 [6.2]%; P = 0.005), largely due to a reduction in hyperglycaemia (>10 mM/L, or 180 mg/dL: median [MAD] 1.9 [2.2]% vs. 3.9 [3.6]%; P = 0.02). CONCLUSIONS: Real-time CGM data and alarms are associated with reductions in low sensor glucose, elevated sensor glucose, and glycaemic variability. This suggests CGM allows patients to detect hyperglycaemic peaks and imminent hypoglycaemia, allowing dietary modification and self-treatment to reduce hypoglycaemia. The use of CGM devices may improve safety in PBH, particularly for patients with hypoglycaemia unawareness.

Insights from Koala-Cattle Interaction Experiments: Koalas and Cattle May See Each Other as a Disturbance.

Koalas are facing many threats and have now been officially listed as endangered. Recently, concerns were raised in anecdotal reports of koalas being killed by livestock, especially cattle. We investigated the significance of cattle as a threat to koala survival via two koala-cattle interaction experiments, from both the koala and cattle perspectives. In the first experiment, we recorded the ranging behaviour of free-ranging, radio-collared koalas prior to, during and after cattle grazed within their usual home range. Koalas decreased their distance travelled and the size of their home range when they shared space with cattle, compared with the period before cattle started grazing within their home range. In the second experiment, we recorded the reactions of cattle towards koalas that they encountered on the ground, using motorised animal models: a model koala mounted on a remote-controlled vehicle and a model dog mounted on the same vehicle, and the vehicle alone. The koala model elicited aggression and fear in cattle, similar to the dog model, whereas their reaction to the vehicle was significantly less aggressive. No actual attacks by the cattle were observed. The results provide experimental evidence that negative koala-livestock interactions occur and indicate that cattle and koalas may see each other as a disturbance.

Activating Human Adipose Tissue with the ß3-Adrenergic Agonist Mirabegron.

An appealing strategy for treatment of metabolic disease in humans is activation of brown adipose tissue (BAT), a thermogenic organ best visualized through 18F-FDG PET/CT. BAT has been activated to varying degrees by mild cold exposure. However, this approach can cause undesirable stress, and there remains no consensus protocol. Here, we describe standardized methods for both acute and chronic activation of BAT using the orally administered ß3-adrenergic receptor (AR) agonist, mirabegron. Acute pharmacological stimulation has enabled quantification of whole-body BAT volume and metabolic activity using PET/CT imaging, and chronic stimulation increases these properties of BAT over time.

Movement of Free-Ranging Koalas in Response to Male Vocalisation Playbacks.

Effective conservation strategies rely on knowledge of seasonal and social drivers of animal behaviour. Koalas are generally solitary and their social arrangement appears to rely on vocal and chemical signalling. Male koala vocalisations, known as bellows, are believed to be closely related to their breeding behaviour. Previous research suggests that oestrous female koalas use bellows to locate unique males to mate with, and that males can similarly use bellows to evaluate the physical attributes of their peers. We tested the behavioural responses of 20 free ranging koalas to bellow recordings collected from small (<6 kg) and large (>8.5 kg) adult male koalas. Individual koala movement was reported by hourly-uploaded GPS coordinates. We report evidence of intra-male competition, with adult males approaching bellow playbacks, particularly those from small-sized males. In contrast, males under three years of age were averse to the playbacks. No patterns in the response of females were detected. Our results provide the strongest evidence yet that bellows are primarily a means by which males occupy and control space during the breeding season. Future studies are required to see if female response to bellows depends on their reproductive status.

Do Livestock Injure and Kill Koalas? Insights from Wildlife Hospital and Rescue Group Admissions and an Online Survey of Livestock-Koala Conflicts.

Koala populations in Australia are declining due to threats such as chlamydiosis, wild dog predation and vehicle collision. In the last decade, grazing livestock emerged anecdotally as a threat to koala survival in areas where koala habitat and livestock grazing land overlap. This is the first study investigating the significance of livestock-inflicted injuries and deaths in koala populations over a large spatial and temporal scale. We investigated the outcome, scale, and frequency of livestock-koala incidents via an online survey and analysed koala admission records in Queensland wildlife hospitals and a wildlife rescue group (Wildlife Victoria) in Victoria. The results provide evidence of both livestock-inflicted injuries and deaths to koalas, especially as these have been confirmed by witness statements. The outcomes for the koala victims of the incidents were severe with a 75% mortality rate. The reported frequency of livestock-koala incidents was low but increasing, with 72 cases (0.14% out of 50,873 admissions) in Queensland wildlife hospitals during 1997-2019, and 59 cases (0.8% of 7017 rescue records) in Wildlife Victoria during 2007-2019. These incidents were likely to be under-reported due to the remoteness of the incident location, possible mis-diagnoses by veterinarians and the possible reluctance of farmers to report them. Future research is encouraged to explore the mechanics and causes of livestock-koala incidents and to develop management strategies to minimise the livestock threat to koalas.

Identification, Genetic Characterization and Validation of Highly Diverse HIV-1 Viruses for Reference Panel Development.

The continued diversification of HIV poses potentially significant challenges to HIV diagnostics and therapeutics. The dynamic evolution of emerging variants is highlighted in countries such as Cameroon in West Central Africa, where all known subtypes and circulating recombinant forms (CRFs) have been shown to be prevalent. We obtained several hundred HIV-positive plasma and viruses from this region for characterization and identification of highly divergent HIV strains. A total of 163 viral strains were cultured to high titers and high volumes using donor peripheral blood mononuclear cells (PBMCs). Initially, 101 viruses representing 59 strains were well characterized and categorized. Results showed that the viral load (VL) range was 0.36-398.9 × 107 copies/mL, p24 values was 0.2-1134 ng/mL. Phylogenetic analysis of thirty-six near full-length HIV-1 genomic sequences demonstrated that most recombinants were highly diverse CRF02 containing unique recombinant forms (URFs). There were seven viral isolates identified as pure subtype/sub-subtypes (F2, A1, G, and D), six as CRFs (CRF06, CRF18, and CRF22), and ten as URFs. These extensively characterized reagents reflect the current dynamic and complex HIV epidemic in Cameroon and provide valuable insights into the potential phylogenetic evolutionary trend of global HIV molecular epidemiology in the future. These materials may be useful for development of HIV validation and reference panels to evaluate the performance of serologic antigen and nucleic acid assays for their ability to detect and quantitate highly divergent HIV strains.

Point-of-care ultrasonography for the diagnosis of testicular torsion: a practical resident curriculum.

Background: Prompt Doppler ultrasonography to aid in diagnosis is often key to managing testicular torsion, but there may be delays in access; a faster, more widely available alternative is point-of-care ultrasonography (POCUS). The purpose of this study was to develop and evaluate a scrotal POCUS curriculum for urology and emergency medicine residents. Methods: Content experts in urology, emergency medicine and diagnostic imaging collaborated in a modified Delphi method to design a practical didactic curriculum for scrotal POCUS for the identification of testicular torsion. Training included 3 online video teaching modules and a 1-hour hands-on teaching session with standardized adult patients. We evaluated participants' competency in scrotal POCUS using a validated scale. We assessed participants' knowledge, comfort and confidence in performing scrotal POCUS before and after the intervention and at 3 months. Results: Twenty-four urology (n = 12) and emergency medicine (n = 12) residents participated in the curriculum. After hands-on practice, 23 participants (96%) were deemed competent at scrotal POCUS. Pre-post testing showed significant improvement in knowledge (mean score 63% v. 80%, p < 0.001), comfort (mean Likert score 0.6 v. 3.6, p < 0.001) and confidence (mean Likert score 1.0 v. 2.1, p < 0.001) after the intervention. These effects were maintained at the 3-month assessment. Conclusion: The scrotal POCUS curriculum was effective and acceptable to both urology and emergency medicine residents. The findings suggest that scrotal POCUS can be learned effectively through a short hands-on session and didactic instruction.

Contexte: Le diagnostic rapide d'une torsion testiculaire à l'aide d'une échographie Doppler est souvent crucial à la prise en charge de ce trouble. Or, comme l'accès à cet examen peut être limité, l'échographie portable constitue une solution de rechange rapide et largement accessible. La présente étude visait la mise au point et l'évaluation d'un programme de formation sur l'échographie portable scrotale destiné aux résidents en urologie et en médecine d'urgence. Méthodes: Des experts en urologie, en médecine d'urgence et en imagerie diagnostique se sont servis d'une méthode Delphi modifiée pour concevoir un programme de formation sur l'échographie portable visant à faciliter le diagnostic de la torsion testiculaire. Ce programme comprenait 3 modules d'apprentissage vidéo en ligne, ainsi qu'une séance pratique d'une heure auprès de patients adultes normalisés. Nous avons évalué les compétences des participants en matière d'échographie portable scrotale au moyen d'une échelle validée. Nous avons également évalué les connaissances, l'aisance et le niveau de confiance des participants à l'égard de cet examen avant et immédiatement après la formation, puis 3 mois plus tard. Résultats: Au total, 24 résidents en urologie (n = 12) et en médecine d'urgence (n = 12) ont suivi le programme de formation. Après la séance pratique, 23 participants (96 %) avaient les compétences nécessaires à la réalisation d'une échographie portable scrotale. La comparaison des résultats obtenus avant et immédiatement après la formation a montré une augmentation significative des connaissances (note moyenne : 63 % c. 80 %; p < 0,001), de l'aisance (moyenne à l'échelle de Likert : 0,6 c. 3,6; p < 0,001) et du niveau de confiance (moyenne à l'échelle de Likert : 1,0 c. 2,1; p < 0,001) des participants. Les effets de la formation étaient toujours présents 3 mois plus tard. Conclusion: Le programme de formation sur l'échographie portable scrotale s'est avéré efficace et acceptable pour les résidents en urologie et en médecine d'urgence. Les résultats obtenus laissent croire qu'une formation pédagogique et une courte séance d'apprentissage pratique permettent l'enseignement efficace de l'échographie portable scrotale.

Conformational triggers associated with influenza matrix protein 1 polymerization.

A central role for the influenza matrix protein 1 (M1) is to form a polymeric coat on the inner leaflet of the host membrane that ultimately provides shape and stability to the virion. M1 polymerizes upon binding membranes, but triggers for conversion of M1 from a water-soluble component of the nucleus and cytosol into an oligomer at the membrane surface are unknown. While full-length M1 is required for virus viability, the N-terminal domain (M1NT) retains membrane binding and pH-dependent oligomerization. We studied the structural plasticity and oligomerization of M1NT in solution using NMR spectroscopy. We show that the isolated domain can be induced by sterol-containing compounds to undergo a conformational change and self-associate in a pH-dependent manner consistent with the stacked dimer oligomeric interface. Surface-exposed residues at one of the stacked dimer interfaces are most sensitive to sterols. Several perturbed residues are at the interface between the N-terminal subdomains and are also perturbed by changes in pH. The effects of sterols appear to be indirect and most likely mediated by reduction in water activity. The local changes are centered on strictly conserved residues and consistent with a priming of the N-terminal domain for polymerization. We hypothesize that M1NT is sensitive to changes in the aqueous environment and that this sensitivity is part of a mechanism for restricting polymerization to the membrane surface. Structural models combined with information from chemical shift perturbations indicate mechanisms by which conformational changes can be transmitted from one polymerization interface to the other.

Harnessing synthetic biology to enhance heterologous protein expression.

The ability to express heterologous proteins in microbial hosts is crucial for many areas of research and technology. In most cases, however, successful expression and purification of the desired protein require fusion to another protein. To date, all fusion partners have been chosen from natural sequences, which evolved for other purposes, and may not be optimal fusion partners. However, the rise of synthetic biology and protein design make it possible to design and optimize fusion proteins using novel sequences that did not arise in nature. Here, we describe a series of De novo Expression Enhancer Proteins (DEEPs) that facilitate high-level expression and facile purification of heterologous proteins and peptides. To test the DEEP system, a de novo protein was fused to several target proteins covering a range of sizes and solubilities. In all cases, fusions to DEEP outperformed fusions to SUMO, a commonly used natural fusion partner. The availability of novel proteins that can be engineered for specific fusion applications could be beneficial to enhance the expression of a wide range of heterologous proteins.

Trends of gallbladder cancer incidence, mortality, and diagnostic approach in urban Shanghai between 1973 and 2009.

PURPOSE: To describe and interpret secular time trends in gallbladder cancer (GBC) incidence, mortality, and diagnostic approach using 37 years of cancer registry data in urban Shanghai. METHODS: Data on registration of GBC in urban Shanghai during 1973 and 2009 were collected by the Shanghai Cancer Registry. To describe time trends and to identify specific time points when significant changes occurred, we used joinpoint regression analysis. RESULTS: The age-standardized rates (ASRs) of incidence increased from 1.1/100,000 (1973-1975) to 2.9/100,000 (2006-2009) in men and from 1.7/100,000 (1973-1975) to 3.9/100,000 (2006-2009) in women. ASRs of incidence increased significantly with estimated annual percent changes (EAPCs) of 2.8% in men and 2.5% in women. The mortality trends increased significantly, with EAPCs of 2.8% in men and 2.5% in women. The increasing incidence and mortality rates were primarily observed in men ⩾60 years of age and in women ⩾70 years of age. Notable downward trends in incidence and mortality were identified among women age 60-69 years over the last decade. The percentage of GBC diagnosed by pathology increased steadily over the years while the percentage of GBC diagnosed by imaging, surgery, and biochemistry sharply increased from 1987 onwards. CONCLUSIONS: Thirty-seven years of cancer registry data document a tremendous increase in incidence/mortality and a slight decline in incidence/mortality over the last decades for GBC, especially among women, in Shanghai. The development of diagnostic approaches and aging population may play important roles.

Characterizing drug-related adverse events by joint analysis of biomedical and genomic data: A case study of drug-induced pulmonary fibrosis.

Spontaneous reporting systems such as the FDA's adverse event reporting system (FAERS) present a great resource to mine for and analyze real-world medication usage. Our study is based on a central premise that FAERS captures unsuspected drug-related adverse events (AEs). Since drug-related AEs result for several reasons, no single approach will be able to predict the entire gamut of AEs. A fundamental premise of systems biology is that a full understanding of a biological process or phenotype (e.g., drug-related AE) requires that all the individual elements be studied in conjunction with one another. We therefore hypothesize that integrative analysis of FAERS-based drug-related AEs with the transcriptional signatures from disease models and drug treatments can lead to the generation of unbiased hypotheses for drug-induced AE-modulating mechanisms of action as well as drug combinations that may target those mechanisms. We test this hypothesis using drug-induced pulmonary fibrosis (DIPF) as a proof-of-concept study.

Golga5 is dispensable for mouse embryonic development and postnatal survival.

Golgins are a family of coiled-coil proteins located at the cytoplasmic surface of the Golgi apparatus and have been implicated in maintaining Golgi structural integrity through acting as tethering factors for retrograde vesicle transport. Whereas knockdown of several individual golgins in cultured cells caused Golgi fragmentation and disruption of vesicle trafficking, analysis of mutant mouse models lacking individual golgins have discovered tissue-specific developmental functions. Recently, homozygous loss of function of GOLGA2, of which previous in vitro studies suggested an essential role in maintenance of Golgi structure and in mitosis, has been associated with a neuromuscular disorder in human patients, which highlights the need for understanding the developmental roles of the golgins in vivo. We report here generation of Golga5-deficient mice using CRISPR/Cas9-mediated genome editing. Although knockdown studies in cultured cells have implicated Golga5 in maintenance of Golgi organization, we show that Golga5 is not required for mouse embryonic development, postnatal survival, or fertility. Moreover, whereas Golga5 is structurally closely related to Golgb1, we show that inactivation of Golga5 does not enhance the severity of developmental defects in Golgb1-deficient mice. The Golga5-deficient mice enable further investigation of the roles and functional specificity of golgins in development and diseases.

The microRNA miR-33a suppresses IL-6-induced tumor progression by binding Twist in gallbladder cancer.

Cytokine is a key molecular link between chronic inflammation and gallbladder cancer (GBC) progression. The potential mechanism of cytokine-associated modulation of microRNAs (miRNAs) expression in GBC progression is not fully understood. In this study, we investigated the biological effects and prognostic significance of interleukin-6 (IL-6) -induced miRNAs in the development of GBC. We identify that inflammatory cytokine, IL-6 promotes proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of GBC both in vitro and in vivo. Among all the changed miRNAs in miRNA profiling, miR-33a expression was significantly decreased in IL-6 treated GBC cell lines, as well as in GBC tissues compared with case-matched normal tissues and cholecystitis tissues. In turn, miR-33a suppresses IL-6-induced tumor metastasis by directly binding Twist which was identified as an EMT marker. High expression of miR-33a suppressed xenograft tumor growth and dissemination in nude mice. The downregulation of miR-33a was closely associated with advanced clinical stage, lymph node metastasis, and poor clinical outcomes in patients with GBC. miR-33a acts as a tumor suppressor miRNA in GBC progression and may be considered for the development of potential therapeutics against GBC.

Expression of interleukin-6 is associated with epithelial-mesenchymal transition and survival rates in gallbladder cancer.

The present study aimed to investigate the expression of interleukin­6 (IL­6) in gallbladder cancer (GBC) tissues and its correlation with survival rate. The association between IL­6 and epithelial­mesenchymal transition (EMT)­associated markers was also examined. Using immunohistochemistry, reverse transcription quantitative polymerase chain reaction (RT­qPCR) and western blot analysis, the protein and mRNA expression levels of IL­6, Twist, E­cadherin and Vimentin in 20 GBC tissues were analyzed. The IL­6, Twist and Vimentin proteins were overexpressed in 40, 20 and 70% of the human GBC samples, respectively. The protein expression of E­cadherin was higher in only 5% of the GBC samples. These differences were significant (P<0.05). Western blot analysis also revealed overexpression of IL­6, Twist and Vimentin and underexpression of E­cadherin in the GBC samples with poor differentiation, local invasion and a higher tumor­node­metastasis (TNM) stage (P<0.05). Higher mRNA expression levels of IL­6, Twist and Vimentin and a reduced expression level of E­cadherin were also demonstrated in the GBC tissues (P<0.05). The degree of differentiation, local invasion, lymph node metastasis and clinical stage were significantly associated with the mRNA expression levels of IL­6, Twist and E­cadherin. The increased expression levels of IL­6 and Twist and the reduced expression of E­cadherin correlated with shorter median survival rates (P<0.05). Line regression results revealed correlation among the mRNA expression levels of IL­6, Twist, E­cadherin and Vimentin. To the best of our knowledge, the present study is the first to demonstrate that IL­6 is associated with EMT­associated markers, tumor differentiation, local invasion, TNM stage and survival rates in GBC.

Synergic silencing of costimulatory molecules prevents cardiac allograft rejection.

BACKGROUND: While substantial progress has been made in blocking acute transplant rejection with the advent of immune suppressive drugs, chronic rejection, mediated primarily by recipient antigen presentation, remains a formidable problem in clinical transplantation. We hypothesized that blocking co-stimulatory pathways in the recipient by induction of RNA interference using small interference RNA (siRNA) expression vectors can prolong allogeneic heart graft survival. METHOD: Vectors expressing siRNA specifically targeting CD40 and CD80 were prepared. Recipients (BALB/c mice) were treated with CD40 and/or CD80 siRNA expression vectors via hydrodynamic injection. Control groups were injected with a scrambled siRNA vector and sham treatment (PBS). After treatment, a fully MHC-mismatched (BALB/c to C57/BL6) heart transplantation was performed. RESULT: Allogeneic heart graft survival (>100 days) was approximately 70% in the mice treated simultaneously with CD40 and CD80 siRNA expression vectors with overall reduction in lymphocyte interstitium infiltration, vascular obstruction, and edema. Hearts transplanted into CD40 or CD80 siRNA vector-treated recipients had an increased graft survival time compared to negative control groups, but did not survive longer than 40 days. In contrast, allogenic hearts transplanted into recipients treated with scrambled siRNA vector and PBS stopped beating within 10-16 days. Real-time PCR (RT-PCR) and flow cytometric analysis showed an upregulation of FoxP3 expression in spleen lymphocytes and a concurrent downregulation of CD40 and CD80 expression in splenic dendritic cells of siRNA-treated mice. Functional suppressive activity of splenic dendritic cells (DCs) isolated from tolerant recipients was demonstrated in a mixed lymphocyte reaction (MLR). Furthermore, DCs isolated from CD40- and CD80-treated recipients promoted CD4+CD25+FoxP3+ regulatory T cell differentiation in vitro. CONCLUSION: This study demonstrates that the simultaneous silencing of CD40 and CD80 genes has synergistic effects in preventing allograft rejection, and may therefore have therapeutic potential in clinical transplantation.

Simultaneous pancreas-kidney transplantation: The role in the treatment of type 1 diabetes and end-stage renal disease.

Type 1 diabetes mellitus (DM) is one of the most common and debilitating diseases to affect the world. Many patients are afflicted by microvascular and macrovascular complications, and succumb to end-stage renal disease (ESRD). Although dialysis and insulin therapy provides better glycemic control, it nonetheless significantly decreases a patient's quality of life. Moreover, they cannot reverse ESRD or alleviate complications. Simultaneous pancreas-kidney (SPK) transplantation has revolutionized the way we manage type 1 DM; it provides a physiological means of achieving normoglycemia while rendering patients free of dialysis. Understanding this procedure is important because it is becoming a more common management strategy for patients with type 1 DM. In this review, we will begin with a brief summary of type 1 DM, followed by a comprehensive description of SPK procedure, including the history and technique. We will then present the outcomes of transplantation.