Infiltration Of Liver Research Articles

Acute toxicities of intravenous, intraperitoneal, or intratumoral injection of natural killer cells in human pancreatic adenocarcinoma-bearing mice: Randomized study

AimsTo investigate the possible acute toxicities and pathological changes associated with intravenous, intraperitoneal, or intratumoral injection of natural killer (NK) cells in mice subcutaneously bearing human pancreatic adenocarcinoma (PaC). Methods100 NPG tumor-bearing mice (50/sex) were engrafted subcutaneously with human PaC BXPC-3 cells 9 days before administration. They were randomly divided into 10 groups with 5 males and 5 females in each group. Mice in Group 1 were given sodium chloride intravenously as vehicle control, and mice in Groups 2–4 human peripheral blood-derived NK cells intravenously at doses of 2 × 107, 1 × 108, and 5 × 108 cells/kg, respectively; mice in Groups 5–7 were injected with NK cells intraperitoneally at doses of 2 × 107, 1 × 108, and 5 × 108 cells/kg, respectively, and mice in Groups 8–10 with NK cells intratumorally at doses of 4 × 103, 2 × 104, and 1 × 105 cells/mm3, respectively. Each group was given a single dose; the mice were observed clinically, and body weight, food intake, blood biochemistry, and tumor volume were measured. On Day 15, the mice were euthanized for gross anatomy and histopathology. ResultsOn planned euthanasia, in Groups 2–4 no gross or microscopic pathological changes related to cells injection were found; in Groups 5–7 mice of both sexes showed a decrease in extramedullary hematopoiesis of spleen, and at the dose of 5 × 108 cells/kg, mice of both sexes showed an increase in the composition of spleen white pulp cells. In Groups 8–10, mice of both sexes at doses of 4 × 103 and 1 × 105 cells/mm3 and female mice at the dose of 2 × 104 cells/mm3 showed a decrease in extramedullary hematopoiesis of spleen, and female mice at a dose of 4 × 103 cells/mm3 and mice of both sexes at doses of ≥ 2 × 104 cells/mm3 showed an increase in the composition of spleen white pulp cells; perivascular/peribronchiolar inflammatory cell infiltration in lung and bronchus was observed in mice of both sexes at doses of ≥ 2 × 104 cells/mm3, and inflammatory cell infiltration in liver was observed in mice of both sexes at a dose of 1 × 105 cells/mm3. No other abnormal changes with toxicological significance in clinical observation, body weight, food intake, or blood biochemistry were observed in each group. ConclusionsIn our study intravenous injection appears the safest way to give NK cells to human PaC-bearing mice. Using intraperitoneal or intratumoral administration, spleen, liver, and lung were the most often affected organs, albeit with mostly mild pathological changes.

Regulatory Effects and Mechanisms of L-Theanine on Neurotransmitters via Liver-Brain Axis Under a High Protein Diet.

Excessive protein intake causes liver and brain damage and neurotransmitter disorders, thereby inducing cognitive dysfunction. L-theanine can regulate the neurotransmitter content and show great potential in liver and brain protection. However, it remains unclear whether l-theanine effectively regulates neurotransmitter content under high-protein diet. A 40-day feeding experiment was performed in Sprague Dawley rats to investigate the regulatory effects and mechanisms of l-theanine on neurotransmitters via liver-brain axis in high-protein diets. The results showed that a 30% protein diet increased the liver and brain neurotransmitter content while maintaining the normal structure of liver and the hippocampal CA1 of brain and improving the autonomous behavior of rats. In contrast, 40% and 50% protein diets decreased the content of neurotransmitters, affected autonomous behavior, destroyed the hippocampal CA1 of brain structure, increased hepatic inflammatory infiltration, lipid degeneration, and hepatocyte eosinophilic change in liver, increased liver AST, ALT, MDA, CRP, and blood ammonia level, and decreased liver SOD and CAT level. However, l-theanine improved liver and brain neurotransmitter content, autonomous behavior, liver and hippocampal brain structure, and liver biochemical indicators in 40% and 50% protein diets. To explore how LTA can eliminate the adverse effects of a high-protein diet, we analyzed different metabolites and proteomes and using western blotting for validate quantitatively. We found that l-theanine regulates the activity of PF4 and G protein subunit alpha i2, increases the content of brain-derived neurotrophic factor and dopamine under a 20% protein diet. In addition, l-theanine can activate the adenylate cyclase-protein kinase A pathway through the protein alpha/beta-hydrolase domain protein 12 to regulate the content of neurotransmitters under a 40% protein diet, thereby exerting a neuroprotective effect.

Antitumor activity of a whey peptide-based enteral diet in C26 colon tumor-bearing mice.

The antitumor effects of a whey peptide-based enteral diet, whose main components are whey peptides and yogurt fermented by Lactobacillus delbureckii subsp. bulgaricus 2038 and Streptococcus thermophilus 1131, were investigated in mice. Our results indicated that the tumor weight in C26 carcinoma-transplanted mice was significantly smaller at day 16 post-implantation in the whey peptide-based enteral diet group (1.36±0.54g) than in the control group (1.83±0.89g) (p <0.05). The whey peptide-based enteral diet group exhibited higher tumor cell apoptosis, lower cell proliferation, and inactive angiogenesis indicating by higher degree of TUNEL, lower positive rates of Ki-67, VEGF, and CD34 than control group. It also attenuated inflammatory cell infiltration of spleen and liver as indicated by the decreased spleen index (10.89±2.06vs. 12.85±2.92, p<0.05) and increased liver index (58.09±11.37vs. 53.19±6.67, p<0.05) in the whey peptide-based enteral diet group than the control diet group. These results proved the inhibitory effect of the whey peptide-based enteral diet on tumor growth, which might be attributed to the whey peptides component. PRACTICAL APPLICATION: A whey peptide-based enteral diet (MEIN® ), containing cheese whey and multiple nutrients, was selected to verify the anti-tumor effect by animal experiments. The tumor weight growth, tumor cell proliferation, inflammatory cell infiltration of spleen and liver in tumor model mice was significantly attenuated by the whey peptide-based enteral diet, that might be attributed to its whey peptides component. These results provided an additive direction for cancer therapy and need a further study including clinical trials.

Biochemical, Hematological and Histopathological Alterations in Liver and Lung of Rats Due to Sub-chronic Inhalation Exposure to NPK 15.15.15 Fertilizer

The current study has been conducted to evaluate biochemical, hematological and histopathological alterations in rats following one month of inhalation exposure to NPK (nitrate, phosphate, potassium) 15.15.15 fertilizer. 24 male Wistar rats were randomly divided into four groups (A, B, C and D). Group A was served as control, whereas B, C and D groups have been exposed to aerosol of different doses (10 mg/m3, 30 mg/m3 and 50 mg/m3, respectively) of NPK 15.15.15 fertilizer during 30 min/day for one month. At the end of the experiment, rats were sacrificed. Blood samples were collected to evaluate biochemical and hematological parameters. Liver and lung tissues were served to evaluate oxidative stress statue and histopathological alterations.&#x0D; Our results showed that inhalation exposure of rats to NPK 15.15.15 fertilizers, especially at high dose (50 mg/m3) resulted in a significant (P = 0,05) elevation of biochemical hepatic markers: Aspartate aminotransferase and Alanine transaminase, and a highly significant (P = 0,01) elevation of alkaline phosphatase. In addition, there were significant changes in hematological parameters revealed mainly by a highly significant (P = 0,01) increase of lymphocytes in both groups C and D groups, and significant decreases (P = 0,05) in all of: red blood cells, hematocrit, and hemoglobin in rats of group D compared to control. Furthermore, the inhalation exposure of rats to NPK 15.15.15 fertilizers induced lung oxidative stress revealed by the reduction of anti-oxidant enzymes (Glutathione peroxidase, reduced glutathione and superoxide dismutase); besides we have observed significant histopathological alterations revealed mainly by inflammatory cell infiltration in liver and lung tissues.&#x0D; The inhalation exposure to important doses of NPK 15.15.15 fertilizer can damage the liver and lung tissues by altering biochemical, hematological and oxidative stress parameters.

Pathological characteristics and congenital immunological responses of pigeons-infected with Neospora caninum

Pigeons are natural intermediate host of Neospora caninum (N. caninum). In comparison to ruminants, N. caninum causes milder clinical symptoms and less financial loss to pigeons. Natural infectious rates and high prevalence of N. caninum in pigeons, and death cases of N. caninum-infected pigeons under experimental conditions have been reported, but the detailed pathological characteristics and congenital immunological responses of pigeons-infected with N. caninum remain not well described. In this study, pigeons were infected intraperitoneally with 107 N. caninum tachyzoites. N. caninum in tissues was detected by qPCR. Pathological changes of tissues were examined by hematoxylin-eosin staining. Blood smears were prepared for counting eosinophils changes in blood. Heterophil extracellular traps (HETs) in vivo and in vitro were quantified by Pico Green. N. caninum-induced HETs structures were observed by immunofluorescence staining. The model of pigeons-infected with N. caninum was successfully established. Lung and duodenum were the main target organs of pigeons-infected with N. caninum. N. caninum caused hemorrhage, edema and inflammatory cell infiltration in liver, pulmonary congestion and hemorrhage, organizational destruction in lung, and shorter villi or even disappear in duodenum. N. caninum also increased the number of eosinophils in blood of pigeons. Moreover, N. caninum-induced HETs release in the congenital immunological system of pigeons were first demonstrated, and the HETs structures were consisted of DNA as the skeleton and modified with citH3 and elastase. N. caninum-induced HETs release was related with NADPH oxidase, TLR 2 and 4, ERK1/2 and p38 MAPK signaling pathways, and glycolysis. In summary, it is the first report on the detailed pathological characteristics and congenital immunological responses of pigeons-infected with N. caninum, which may provide theoretical basis for the prevention and control of Neosporosis in pigeons.

Gemcitabine, Cisplatin, and Nab-Paclitaxel as a First-Line Therapy for Advanced Biliary Tract Cancers.

Locally advanced, inoperable, or metastatic gallbladder cancers (GBC) are treated with either gemcitabine-platinum combinations or gemcitabine alone based on physician discretion. However, the combination of gemcitabine, cisplatin, and nab-paclitaxel (GCNP) has shown increased response rates and prolonged survival in a phase II trial of biliary tract patients. Consecutive series of patients diagnosed with locally advanced (liver infiltration > 5cm, large nodes at porta, abutting duodenum), inoperable, and metastatic biliary tract patients between January 2018 and August 2022 were evaluated for first-line chemotherapy GCNP, in the multidisciplinary joint clinic (MDJC). The primary endpoint was ORR, and the major secondary endpoint was event-free survival (EFS). A total of 142 patients received GCNP during the specified time period. The median age of the cohort was 52years (range: 21-79), the majority were females (61.3%), and the majority were GB (81.7%). Response rates were available in 137 patients. Complete response, partial response, and stable disease were seen in 9 (6.3%), 87 (61.3%), and 24 (16.9%), respectively, for an ORR of 67.6% and a clinical benefit rate of 84.5%. The median EFS was 9.92 (95% CI, 7.69-12.14) months. Of the 52 patients in whom GCNP was given with NACT intent for locally advanced GBC, 17 patients underwent surgery (34%). Our study indicates that GCNP leads to improved response rates, increased chances of resectability, and possibly better survival in patients with GBC.

1574-P: FoxO1 Links Insulin Resistance to Hepatic Inflammation and Catalyzes the Evolution of Benign Steatosis to Nonalcoholic Steatohepatitis

Hepatic inflammation is intertwined with insulin resistance and is a causative factor for the transition of benign steatosis to nonalcoholic steatohepatitis (NASH). We found that FoxO1 was markedly upregulated in hepatic macrophages, coinciding with hepatic inflammation and steatosis in dietary obese mice. We generated myeloid-conditional FoxO1-transgenic and FoxO1-knockout mice, two complementary models for determining the effect of FoxO1 gain- vs. loss-of-function on macrophage activation in response to insulin resistance. We found that myeloid FoxO1 gain-of-function aggravated hepatic inflammation in mice in response to overnutrition or endotoxin. In contrast, myeloid FoxO1 loss-of-function skewed macrophage polarization from pro-inflammatory M1 to anti-inflammatory M2 phenotypes, accompanied by the reduction of macrophage infiltration in liver. This effect contributed to the suppression of hepatic inflammation and improvement of insulin sensitivity. Despite fat-induced obesity, myeloid-conditional FoxO1-deficient mice were protected from developing hyperglycemia, hyperinsulinemia and glucose intolerance. Furthermore, the improvement in hepatic inflammation and insulin resistance translated into a significant beneficial effect on NASH, culminating in the reduction of hepatic steatosis and fibrosis in myeloid FoxO1-deficient mice on NASH-inducing diet. Human FOXO1 expression was upregulated in inflammatory macrophages in liver, correlating with hepatic inflammation, steatosis and fibrosis in patients with NASH. Mechanistically, FoxO1 counteracts Stat6 to skew macrophage polarization from M2 toward M1 signatures with pro-inflammatory profiles. We concluded that myeloid FoxO1 dysregulation is culpable for linking insulin resistance to abnormal macrophage activation. This effect perpetuates hepatic inflammation and catalyzes the evolution of simple steatosis to NASH. Disclosure S.Lee: None. T.Usman: None. G.Chhetri: None. X.Wang: None. H.Dong: None.

Investigating Ineligibility of Potential Living Liver Donors for Transplantation: Experience from a Large Liver Transplant Center in Pakistan

BackgroundLiving Donor Liver Transplantation (LDLT) is common in Pakistan, where the deceased organ donation is lacking. However, not all potential living donors are suitable for the procedure due to various medical, technical, psychosocial, and ethical reasons. This study aims to investigate the reasons for the ineligibility of potential living liver donors in Pakistan. MethodsBetween June 2022 and March 2023, 530 potential living liver donors were assessed for liver transplant recipients at the Pakistan Kidney and Liver Institute & Research center (PKLI&RC). Potential donors were evaluated in three steps, including laboratory tests imaging, and consultations. Prospectively maintained data on all potential donors were examined and retrospectively assessed to identify the causes of donor rejection. ResultsOf the 530 potential living donors evaluated, 364 of the potential donors were considered ineligible (68.67% of donors were rejected). From these rejected donors, 193 (53%) were males, and 171 (47%) were females. The mean age of the rejected donors was 26 years old. The most common reason for disqualifying potential living liver donors was a withdrawal of consent (65 cases, 17.9%). Other factors contributing to ineligibility included fatty liver infiltration reflected by a liver attenuation index (LAI) < 0, future liver remnant (FLR) < 28%, and a graft-to-recipient weight ratio (GRWR) < 0.6. Medical issues, such as abnormal liver function tests and positive hepatitis serology, also resulted in disqualification. ConclusionThis study highlights the importance of meticulous assessment of potential living liver donors to ensure their safety and the success of the transplant. The high rate of withdrawal of consent and the prevalence of medical issues indicates the need for addressing ethical concerns and improving public education on the donation process. This study also emphasizes the importance of a deceased organ donation in developing countries like Pakistan to ensure the availability of organs for transplantation.

Quercetin alleviates gliotoxin-induced duckling tissue injury by inhibiting oxidative stress, inflammation and increasing heterophil extracellular traps release.

Aspergillus fumigatus causes aspergillosis with high morbidity and mortality in the duck industry. As a vital virulence factor produced by A. fumigatus, gliotoxin (GT) is widely present in food and feed, threatening duck industry and human health. Quercetin is a polyphenol flavonoid compound from natural plants with anti-inflammatory and antioxidant functions. However, the effects of quercetin on ducklings with GT poisoning are unknown. The model of ducklings with GT poisoning was established, and the protective effects and molecular mechanisms of quercetin on ducklings with GT poisoning were investigated. Ducklings were divided into control, GT, and quercetin groups. A model of GT (2.5 mg/kg) poisoning in ducklings was successfully established. Quercetin protected GT-induced liver and kidney functions and alleviated GT-induced alveolar wall thickening in lungs, cell fragmentation, and inflammatory cell infiltration in liver and kidney. Quercetin decreased malondialdehyde (MDA) and increased superoxide dismutase (SOD) and catalase (CAT) after GT treatment. Quercetin significantly reduced GT-induced mRNA expression levels of inflammatory factors. Furthermore, quercetin increased GT-reduced heterophil extracellular traps (HETs) in serum. These results indicated that quercetin protected ducklings against GT poisoning by inhibiting oxidative stress, inflammation and increasing HETs release, which confirms the potential applicability of quercetin in treating GT-induced duckling poisoning.

Gemcitabine, cisplatin, nab-paclitaxel: The effectiveness of this regimen for gall bladder cancer.

e16168 Background: Locally Advanced inoperable/metastatic Gallbladder Cancers (GBC) are treated with either GC, GO, or single-agent Gemcitabine based on physician discretion. However, based on about 50% response rates (ORR) and prolonged survival in the phase II trial of biliary tract patients, were treated with Gemcitabine - Cisplatin - Nab-Paclitaxel (GCNP). Methods: Consecutive series of patients diagnosed with locally advanced (liver infiltration &gt; 5 cm, large nodes at porta, abutting duodenum), inoperable and metastatic biliary tract patients between January 2018 to August 2022 were evaluated for first-line chemotherapy GCNP, in the Multidisciplinary Joint Clinic (MDJC). The primary endpoint was ORR and the secondary endpoints were survival and tolerance. Results: A total of 142 patients received GCNP during the specified time period. The median age of the cohort was 52 years (range: 21- 79), the majority were females (61.3%), and the majority were GB (81.7%). Response rates were available in 137 patients. CR, PR, and SD were seen in 9 (6.3%), 87 (61.3%), and 24 (16.9%) respectively, for a RR of 67.6% and CBR of 84.5%. Median EFS was 9.92 (95% CI, 7.69- 12.14) months. Of the 52 patients in whom GCNP was given with NACT intent for locally advanced GBC, 17 patients underwent surgery (34%). Conclusions: Our study indicates that GCNP leads to better response rates, better chances of unresectable disease being surgically feasible at a later date, and possibly offers better survival in the GBC cohort.

Obesity management for the pre–liver transplant and post–liver transplant patient

Obesity management for the pre–liver transplant and post–liver transplant patient

A phase I, randomized, controlled, multicentre trial of isolated hepatic perfusion in combination with ipilimumab and nivolumab in patients with uveal melanoma metastases (the SCANDIUM 2 trial).

9533 Background: Uveal melanoma is a rare disease characterized by liver metastasis and a very poor prognosis. In patients with metastatic UM, a single treatment with isolated hepatic perfusion (IHP) with high dose melphalan has shown response rates of 40%, while immune checkpoint blockade with ipilimumab (3 mg/kg) in combination with nivolumab (1 mg/kg) every third week (IPI3/NIVO1 q3w) for four cycles followed by nivolumab monotherapy has shown response rates of 11-18%. The impact of these treatments on overall survival, especially if combined, is unclear. This phase I trial investigates the safety and tolerability of the combination of IHP and IPI3/NIVO1. Methods: Eligible in this multicenter open, randomized, controlled, phase I trial, were patients with liver dominant metastatic uveal melanoma who had not received previous systemic treatment. Patients were randomized to receive either (Arm A) IHP followed by combination immunotherapy (four cycles IPI3/NIVO1 q3w) or (Arm B) one neoadjuvant cycle of IPI3/NIVO1 prior to IHP followed by three cycles IPI3/NIVO1 q3w. Thereafter, both Arm A and B received monotherapy with nivolumab (480 mg q4w) for up to 1 year. IHP was performed using melphalan 1mg/kg perfused through the liver for 60 minutes under hyperthermia (40°C). The primary endpoint was incidence and severity of adverse events while, secondary endpoints included response according to RECIST v1.1 criteria reported according by the local investigator. Results: A total of 18 patients were included and randomized, nine to Arm A and nine to Arm B. Three patients did not undergo IHP as planned, one due to perioperative complications (Arm A), one due to extensive metastatic liver infiltration ( &gt; 50% liver volume) (Arm B) and one due to encephalitis related to neoadjuvant IPI3/NIVO1 (Arm B). A total of 20 serious adverse events (SAEs) were reported in eleven patients. There were ten SAEs in each arm, with no treatment related deaths. In total, 11 of 18 patients (six in Arm A and five in arm B) did not complete the planned four cycles of IPI3/NIVO1, with a mean of 2.4 cycles in Arm A and 3.0 cycles in Arm B. Response was evaluable in 17 patients, with the best clinical responses reported as three complete responses (18%), four partial responses (24%), seven stable disease (41%) and three progressive disease (18%). The overall response rate was 63% in Arm A (5/8) and 22% in Arm B (2/9). Conclusions: For previously untreated patients with liver dominant metastatic uveal melanoma, treatment with IHP in combination with IPI3/NIVO1 had a high, yet manageable toxicity profile. The efficacy of this combination treatment is encouraging, however, one cycle of neoadjuvant IPI3/NIVO1 given before IHP did not seem beneficial, neither in regards to safety nor efficacy. Clinical trial information: NCT04463368 .

Macrophage-Derived Cathepsin S Remodels the Extracellular Matrix to Promote Liver Fibrogenesis

Liver fibrosis is an intrinsic wound-healing response to chronic injury and the major cause of liver-related morbidity and mortality worldwide. However, no effective diagnostic or therapeutic strategies are available, owing to its poorly characterized molecular etiology. We aimed to elucidate the mechanisms underlying liver fibrogenesis. We performed a quantitative proteomic analysis of clinical fibrotic liver samples to identify dysregulated proteins. Further analyses were performed on the sera of 164 patients with liver fibrosis. Two fibrosis mouse models and several biochemical experiments were used to elucidate liver fibrogenesis. We identified cathepsin S (CTSS) up-regulation as a central node for extracellular matrix remodeling in the human fibrotic liver by proteomic screening. Increased serum CTSS levels efficiently predicted liver fibrosis, even at an early stage. Secreted CTSS cleaved collagen 18A1 at its C-terminus, releasing endostatin peptide, which directly bound to and activated hepatic stellate cells via integrin α5β1 signaling, whereas genetic ablation of Ctss remarkably suppressed liver fibrogenesis via endostatin reduction invivo. Further studies identified macrophages as the main source of hepatic CTSS, and splenectomy effectively attenuated macrophage infiltration and CTSS expression in the fibrotic liver. Pharmacologic inhibition of CTSS ameliorated liver fibrosis progression in the mouse models. CTSS functions as a novel profibrotic factor by remodeling extracellular matrix proteins and may represent a promising target for the diagnosis and treatment of liver fibrosis.

The significant IgG4 infiltrate in autoimmune hepatitis is associated with a greater ductular reaction and more advanced liver disease

BackgroundSclerosing cholangitis is the typical IgG4-related disease digestive involvement. However, the role of the IgG4 liver expression in autoimmune hepatitis remains unknown. Aimsto assess whether the expression of IgG4 plasma cells in patients with autoimmune hepatitis (AIH) was associated with different outcomes. MethodsRetrospective study including patients diagnosed with AIH by biopsy from January-2009 to June-2021. At least mild IgG4 expression (>10 IgG4+-plasma cells per field) was considered as significant. Results85 patients with AIH were included. Overall, 58.8% were women, mean age 54 years. Nine (10.6%) presented cirrhosis at diagnosis. Fifteen (17.6%) had significant IgG4 liver expression. Patients with IgG4 infiltrate were older (p = 0.021), presented liver cirrhosis more frequently (33.3% vs. 5.7%, p = 0.007), greater IgG plasma values (p = 0.008) and atypical ANCAs (p = 0.086); ductular reaction was also more common (p = 0.009). Complete remission rate was similar regardless of the IgG4 infiltrate. Time to corticosteroids discontinuation was longer in subjects with IgG4 infiltrate (p = 0.068), but second-line therapy tended to be less frequent (p = 0.187). ConclusionSignificant IgG4 liver infiltrate in patients with autoimmune hepatitis is associated with more advanced liver disease. The greater ductular reaction mediated by the IgG4 infiltrate may be the cause for this finding, though this finding should be prospectively assessed.

Diagnostic performance and safety of endoscopic ultrasound-guided fine-needle aspiration/biopsy for gallbladder lesions.

Endoscopic ultrasound-guided fine-needle aspiration and fine-needle biopsy (EUS-FNA/FNB) is not fully established as a pathological sampling tool for gallbladder lesions due to limited evidence. We therefore aimed to clarify the effectiveness and safety of this procedure in a large-population cohort. This study retrospectively evaluated the diagnostic yield of EUS-FNA/FNB for accurately differentiating between benign and malignant gallbladder lesions. Puncture targets included the gallbladder mass, lymph node, and liver mass. Adverse events and factors associated with diagnostic accuracy were analyzed as well. In 187 patients with gallbladder lesions undergoing EUS-FNA/FNB, 18 benign lesions and 169 malignant lesions were identified. Overall sampling adequacy was 98% (184/187). The diagnostic accuracy of EUS-FNA/FNB was 97% (182/187), sensitivity was 97% (164/169), and specificity was 100% (18/18). A single postprocedural complication (minor bleeding) was recorded in one patient. In the 169 cases of malignancy, 203 sites were punctured for pathological sampling of the primary mass (n = 94), lymph node (n = 79), and metastatic liver mass (n = 30). No significant difference was found for diagnostic accuracy among the puncture sites (P = 0.70). In cases having specimens obtained from the primary mass, the accuracy of those targeting liver invasion sites was significantly higher than that of other sites (98% vs. 83%, P < 0.01). EUS-FNA/FNB demonstrated clinical usefulness and safety for the pathological diagnosis of gallbladder lesions, with high diagnostic yield and a low incidence of adverse events. Targeting the site of liver infiltration may improve the diagnostic rate of EUS-FNA/FNB in the primary mass.

Prevotella intermedia Aggravates Subclinical Hypothyroidism

Subclinical hypothyroidism (SCH) has been shown to be associated with microbiota. However, the association between SCH and oral microbiota has not yet been elucidated. The results of our previous clinical studies showed that Prevotella intermedia was abundant in the oral microbiota of SCH patients. This study aimed to investigate the relationship between SCH and oral microbiota, verify the pathogenicity of P. intermedia in SCH, and preliminarily explore the possible mechanism. The SCH mouse model with oral application of P. intermedia was established, and the variance in the mouse oral microbiota and changes in thyroid function and metabolism were detected in mice. Student’s t test and analysis of variance were used for statistical analysis. Oral application of P. intermedia changed the composition of the oral microbiota of SCH mice, which enhanced the damage to the thyroid and decreased the expression of functional genes of the thyroid. Moreover, P. intermedia decreased oxygen consumption and aggravated glucose and lipid metabolism disorders in SCH mice. Glucose tolerance and insulin tolerance decreased, and the triglyceride content of the liver and inflammatory infiltration in adipose tissue increased in SCH mice after P. intermedia stimulation. Mechanistically, P. intermedia increased the proportion of CD4+ T cells in cervical lymph nodes and thyroids in SCH mice. Th1 cells were suggested to play an important role in the pathogenesis of SCH involving P. intermedia. In conclusion, P. intermedia aggravated SCH manifestations, including thyroid dysfunction and glucose and lipid metabolism disorders, by causing immune imbalance in mice. This study sheds new light on the pathogenesis of SCH from the perspective of oral microbiota.

Imaging Phenotypes and Evolution of Hepatic Langerhans Cell Histiocytosis on CT/MRI: A Retrospective Study of Clinical Cases and Literature Review.

(1) Background: pathological changes in hepatic Langerhans cell histiocytosis (LCH) have been observed; however, corresponding imaging findings can appear vague to physicians and radiologists. The present study aimed to comprehensively illustrate the imaging findings of hepatic LCH and to investigate the evolution of LCH-associated lesions. (2) Methods: LCH patients with liver involvement treated at our institution were retrospectively reviewed along with prior studies in PubMed. Initial and follow-up computed tomography (CT) and magnetic resonance imaging (MRI) were systematically reviewed, and three imaging phenotypes were created based on the lesion distribution pattern. Clinical features and prognoses were compared among the three phenotypes. Liver fibrosis was evaluated visually on T2-weighted imaging (T2WI) and diffusion-weighted imaging (DWI), and apparent diffusion coefficient (ADC) values of the fibrotic areas were measured. Descriptive statistics and a comparative analysis were used to analyze the data. (3) Results: based on the lesion distribution pattern on CT/MRI scans, patients with liver involvement were categorized as the disseminated lesion phenotype, scattered lesion phenotype, and central periportal lesion phenotype. Patients with scattered lesion phenotype were typically adults, and only a few of them had hepatomegaly (npresent = 1, 1/6, 16.7%) and liver biochemical abnormalities (npresent = 2, 2/6, 33.3%); patients with central periportal lesion phenotype were typically young children, and hepatomegaly and biochemical abnormalities were more apparent in these patients than those with another phenotype; and those with the disseminated lesion phenotype were found in all age groups, and the lesions evolved rapidly on medical imaging. Follow-up MRI scans show more details and can better document the evolution of lesions than CT. T2-hypointense fibrotic changes, including the periportal halo sign (npresent = 2, 2/9, 22.2%), patchy liver parenchyma changes (npresent = 6, 6/9, 66.7%), and giant hepatic nodules close to the central portal vein (npresent = 1, 1/9, 11.1%), were found, while fibrotic changes were not observed in patients with the scattered lesion phenotype. The mean ADC value for the area of liver fibrosis in each patient was lower than the optimal cutoff for significant fibrosis (METAVIR Fibrosis Stage ≥ 2) in a previous study that assessed liver fibrosis in chronic viral hepatitis. (4) Conclusions: The infiltrative lesions and liver fibrosis of hepatic LCH can be well characterized on MRI scans with DWI. The evolution of these lesions was well demonstrated on follow-up MRI scans.

Association of Serum Leptin With Body Mass Index in Gallbladder Cancer Patients: A Pilot Study.

Leptin has been proposed to be a link between obesity and the increased incidence of various cancers like breast cancer, colon cancer, gastric cancer, etc. The role of leptin in gallbladder cancer is largely undetermined. Moreover, no study has evaluated serum leptin levels and their correlation with clinicopathological characteristics and serum tumour markers in gallbladder cancer (GBC). Therefore, the present study was planned. A cross-sectional study was conducted in a tertiary care hospital in Northern India after obtaining ethical approval from the institution. Forty GBC patients staged as per American Joint Committee on Cancer (AJCC) 8th staging systemwere recruited along with 40 healthy controls. Serum leptin was assayed by sandwich enzyme-linked immunosorbent assay (ELISA) and tumour markers (CA19-9, CEA and CA125) by Chemiluminescence.ROC, Mann Whitney U test, Linear regression and Spearman correlation was performed using Statistical Product and Service Solutions (SPSS) (IBM SPSS Statistics for Windows, Version 25.0, Armonk, NY). BMI was also assessed for both groups. Median BMI for GBC patients was 19.46 (IQR 17.61-22.36). Median serum leptin levels were significantly lower (2.09 (IQR 1.01-7.76) ng/mL) in GBC patients as compared to controls (12.32 (IQR 10.50-14.72) ng/mL). AUC was 0.84 with 100% sensitivity and 75% specificity at 7.57 ng/mL.Serum leptin was not associated with cancer stage, resectability, metastasis, liver infiltration, or tumour markers on linear regression (p=0.74, adjusted R square = -0.07). A significant positive correlation was found between BMI and serum leptin in GBC patients (p=0.00). Lower BMI and relatively lean presentation of GBC patients may account for low serum leptin levels.

Phenotypic Evaluation of TREX1 Null Mice as a Model for Lupus-Like Autoimmune Disease

Abstract TREX1 (DNase III) is the major mammalian 3’-5’ DNA exonuclease and thought to play a major role in cell death and genomic DNA degradation. Dysfunction of Trex1 has been suggested to activate immune response by self DNA, as TREX1 null mice develop inflammatory myocarditis similar to autoimmune cardiomyopathy and produce type 1 IFN. TREX1 D18N causes a monogenic cutaneous form of lupus called familial chilblain lupus and exhibits dysfunctioned dsDNA-degrading activity, providing a link between dsDNA degradation and nucleic acid-mediated autoimmune diseases. Using CRISPR-Cas9, we generated a Trex1 −/−mouse by knocking out exon 2 and performed comprehensive phenotypic analyses to evaluate the loss of function of TREX1 and its potential application in SLE research. Both female and male TREX1 null mice showed dramatically increased post-wean morbidity and significantly increased serum ALT, AST and LDH levels, compared to wildtype and heterozygotes from 6 to 24 weeks of age, indicating liver and heart injuries. Pathohistological analysis showed inflammatory immune cell infiltration in liver, heart, spleen, kidney, lung and skin among TREX1 null mice. Serum anti-dsDNA antibody levels were significantly increased and remained high from 7 to 25 weeks of age, consistent with Trex1’s being the major DNA exonuclease. Urine protein to creatinine ratio was dramatically increased at 28 weeks of age among TREX1 null mice. In conclusion, the Trex1 −/−mice we generated in this work showed lupus-like inflammatory autoimmune responses in multiple organs, providing an urgently needed tool to study Trex1-mediated autoimmunity and to evaluate potential therapeutic treatments for lupus.

Immunophenotyping to improve the mechanistic understanding of idiosyncratic drug-induced liver injury: clinical implications and future directions

The late event onset of a fraction of idiosyncratic drug-induced liver injury (DILI) cases and the link observed by genome-wide association studies (GWASs) of certain human leucocyte antigen (HLA) alleles with DILI due to specific drugs support the crucial role of the immune system (both innate and adaptive) in the pathogenesis of DILI. Recent advances in both flow and mass cytometry have allowed the profiling of all major immune cell types in a given sample. Therefore, determining the lymphocyte populations in samples from patients with DILI would facilitate the development of specific biomarkers for DILI diagnosis and prognosis. To date, a few studies have explored the immune landscape in DILI. In a recent study of leukocyte immunophenotyping using flow cytometry from the Spanish DILI Registry, an important role of adaptive immune response in DILI is suggested. DILI patients had significantly higher levels of T helper 1 (Th1) cells and activated helper and cytotoxic T cells than healthy controls. Furthermore, the increased expression of negative immune checkpoints and ligands in DILI patients could reflect a restoration of the immune homeostasis. Differences in the profile of cytokines in DILI patients from the Drug-Induced Liver Injury Network (DILIN) also suggest an involvement of both innate and adaptive immune systems in DILI development and prognosis. Moreover, several studies based on immunophenotyping of liver infiltrates showed a distinctive pattern of cellular infiltrates in patients with immune checkpoint inhibitors (ICIs)-DILI, with lower levels of plasma cells, CD20+ B cells and CD4+ T cells than in autoimmune hepatitis (AIH) patients. These pioneering studies highlight the importance of immunophenotyping for the mechanistic understanding of DILI. In this review, available data on immunophenotyping in DILI are gathered, and the potential clinical applications of cutting-edge, novel immunophenotyping techniques are discussed.