Titanium dioxide nanoparticles (TiO2 NPs), widely used as food additives, frequently coexist with high-fat diets (HD) in modern dietary patterns, yet their combined in vivo toxicity remains poorly understood. This study investigated the multi-organ effects of co-exposure to TiO2 NPs or food-grade E171 and HD in male C57BL/6J mice. Mice were randomly assigned to six groups and fed regular or high-fat diets containing 1 wt% TiO2 NPs or E171 for 13 weeks. Histopathology, serum biochemistry, organ coefficients, and open-field behavioral tests were used to assess tissue injury and functional alterations. Co-exposure to TiO2 NPs and HD markedly exacerbated tissue damage across multiple organs. In the liver, more severe ballooning degeneration, necrosis, and inflammatory infiltration were observed, accompanied by altered liver enzymes and reduced organ coefficients. Intestinal injury was characterized by crypt distortion and increased inflammation, particularly in the HD + TiO2 group. Testicular tissues showed disorganized seminiferous tubules, loss of spermatogenic cells, and interstitial hyperplasia. In the brain, hippocampal neurons exhibited pyknosis and disarray, with decreased brain coefficients and impaired exploratory behavior. E171 induced similar but milder effects. These findings indicate that HD enhances TiO2 NPs induced multi-organ toxicity, highlighting the health risks of realistic co-exposure to dietary nanoparticles and high-fat foods.

Hepatocellular carcinoma (HCC) is a lethal malignancy in which stereotactic body radiotherapy (SBRT) is used for inoperable cases. However, radiation-induced liver disease (RILD) remains a major risk, particularly in fibrotic livers. This study established rat models of RILD with and without preexisting fibrosis to evaluate the effects of radiation-fibrosis on liver damage. Male Sprague-Dawley rats were divided into radiation therapy (RT) (n=41; 25 Gy right liver irradiation) and thioacetamide (TAA)+RT (n=46; 6-week TAA-induced fibrosis + 20 Gy RT) groups. Pathological assessments (Hematoxylin and Eosin, Masson's Trichrome, Picro-Sirius Red, and TGF-β/α-SMA immunohistochemistry) were performed at 2, 4, 8, and 12 weeks post-RT to quantify fibrosis, collagen, inflammation, and ballooning degeneration. Statistical analyses included independent sample t-tests, Mann-Whitney U tests, and one-way ANOVA, with p<0.05 considered significant. The RT group exhibited mild edema (2-12 weeks), mild ballooning degeneration (4-12 weeks), and minimal inflammation (2-12 weeks). Collagen deposition and TGF-β expression increased significantly at 8-12 weeks (p<0.05), whereas α-SMA remained mildly elevated at 4-12 weeks (p>0.05). The TAA+RT group showed mild/severe ballooning degeneration, moderate inflammation, and markedly higher collagen/fibrosis compared with the RT group (p<0.05). Both TGF-β and α-SMA increased progressively in the TAA+RT group, peaking at 12 weeks (p<0.05). Radiation combined with preexisting fibrosis exacerbates hepatic damage and stellate cell activation. This study provides validated RILD models for translational research and highlights the need for cautious radiation dose selection in patients with fibrosis to mitigate the risk of liver injury.

Metabolic dysfunction-associated steatohepatitis (MASH) is the most common chronic liver disease worldwide, with a particularly high incidence among individuals with type 2 diabetes and obesity. Glucagon-like peptide-1 receptor agonists (GLP-1RAs) have emerged as effective weight loss agents, with growing evidence suggesting potential benefits in MASH. This study aimed to evaluate the efficacy and safety of GLP-1RAs in improving the histologic features of MASH. This study is a meta-analysis of randomized controlled trials evaluating the effect of FDA-approved GLP-1RAs in patients with biopsy-confirmed MASH, up to May 2025. The primary outcomes were resolution of MASH without worsening of fibrosis and a ≥1 improvement in fibrosis stage without worsening of MASH at 12-18 months. Secondary outcomes included ≥1-point improvements in the nonalcoholic fatty liver disease activity score, changes in liver histologic features, and the incidence of serious adverse events. A total of 6 randomized controlled trials with 1555 patients (1082 GLP-1RA and 473 placebo) with MASH were included. At 18 months, a significantly greater proportion of patients in the GLP-1RA group achieved MASH resolution without worsening fibrosis compared with the placebo group (OR: 4.16, 95% CI: 2.33-7.42, p<0.001). A subgroup analysis excluding studies with patients with cirrhosis showed a significant benefit on fibrosis regression (OR: 2.02, 95% CI: 1.56-2.62, p=0.01). GLP-1RAs significantly improved nonalcoholic fatty liver disease activity scores and liver histologic features, including balloon degeneration, lobular inflammation, and steatosis, compared with placebo. The pooled serious adverse event rate for GLP-1RAs was comparable to placebo. In patients with noncirrhotic MASH, GLP-1RAs appear to be associated with both MASH resolution without fibrosis worsening and ≥1-stage fibrosis regression without MASH worsening.

Metabolic dysfunction-associated steatohepatitis (MASH), a progressive subtype of metabolic dysfunction-associated steatotic liver disease (MASLD), is characterized by hepatic steatosis, lobular inflammation, and hepatocyte ballooning, and may further progress to liver fibrosis and cirrhosis. Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), a member of the scavenger receptor family, recognizes and binds oxidized low-density lipoprotein. This study aims to investigate the role of LOX-1 in MASH progression. LOX-1 expression in MASLD mouse liver was analyzed using Gene Expression Omnibus (GEO) datasets. Immunofluorescence staining was performed to detect LOX-1 and alpha-smooth muscle actin (α-SMA) levels and co-localization in fibrotic liver tissues and LX-2 cells. LOX-1 knockout (Lox-1-/-) mice were generated using CRISPR/caspase-9 (Cas9) and genotyped by PCR and Sanger sequencing. Wild-type (WT) and Lox-1-/- mice were randomized into control and Western diet model groups. Serum and liver samples were collected for alanine aminotransferase (ALT) and aspartate aminotransferase (AST) measurement by biochemical kits, liver structure evaluation by hematoxylin and eosin (HE) staining, collagen deposition by Masson staining, lipid accumulation by Oil Red O staining, and fibrotic marker gene expression by real-time quantitative PCR (RT-qPCR). Network pharmacology and search tool for the retrieval of interacting genes/proteins (STRING)-based protein-protein interaction (PPI) with Gene Ontology (GO) enrichment were used to predict downstream targets and pathways. The results from the GEO datasets GSE30552 and GSE40041 indicated LOX-1 mRNA was upregulated in high fat diet (HFD) and bile duct ligation (BDL) mouse models (both P<0.001). LOX-1 and α-SMA levels were elevated in fibrotic liver tissues. Lox-1-/- mice were successfully established. Biochemical tests showed that serum AST and ALT levels were significantly elevated in WT mice fed a Western diet (both P<0.001), and these levels decreased after LOX-1 knockout (both P<0.05). HE staining revealed that WT mice on the Western diet exhibited marked hepatocellular ballooning degeneration, steatosis, inflammatory cell infiltration, and periportal fibroplasia, which were significantly ameliorated by LOX-1 knockout. Masson staining demonstrated increased blue-stained collagen fibers in the liver tissues of WT mice fed the Western diet compared with control-diet mice, and LOX-1 knockout inhibited collagen fiber deposition (all P<0.05). RT‑qPCR results showed that hepatic mRNA levels of Acta2, Col1a1, and Timp1 were significantly increased in Western diet-fed mice, and LOX-1 knockout reduced the expression of these fibrogenic marker genes. Oil Red O staining indicated that hepatocytes in WT mice fed the Western diet were notably enlarged, displayed macrovesicular steatosis, and exhibited diffusely distributed red lipid droplets, whereas LOX-1 knockout alleviated hepatic lipid accumulation (both P<0.001). RT‑qPCR results further demonstrated that knockdown of LOX-1 reduced Acta2, Col1a1, and Timp1 mRNA levels in LX‑2 cells (all P<0.05). Immunofluorescence analysis revealed co‑localization of LOX-1 and α‑SMA in LX‑2 cells, and LOX-1 silencing suppressed α‑SMA expression. Network pharmacology suggested LOX-1 may promote MASH via lipid and cholesterol metabolism networks. LOX-1 gene knockout ameliorates Western diet-induced MASH in mice and may serve as a potential therapeutic target.

Nonalcoholic steatohepatitis (NASH) is a chronic liver disease associated with metabolic syndrome, such as obesity, diabetes, and hyperlipidemia, and is a progressive form of nonalcoholic fatty liver disease with pathological features including steatosis, lobular inflammation, and hepatocyte damage (ballooning degeneration). Hepatocellular ballooning degeneration is an important pathological feature of NASH and is closely related to the prognosis of NASH patients. Accurate identification of hepatocellular ballooning degeneration is of great significance for diagnosing NASH and assessing therapeutic response. This article reviews the current status and identification methods of hepatocellular ballooning degeneration in NASH, including invasive and noninvasive.

Tamoxifen (TAM) is one of the most used drugs in the prevention and treatment of breast cancer. A set of common side effects was recorded associating its prolonged clinical use that ranges 3–10 years. This study aimed to investigate TAM-induced parietal cells (PCs) injury in rats and the possible protective effect of rhubarb (Rh) water extract (WE). Twenty-four adult female rats were distributed as: control group, TAM-group (3 mg/kg/day TAM for 4-weeks) and TAM+Rh-group (combined 3 mg/kg/day TAM and 20 mg/kg Rh-WE for 4-weeks). Blood sample before euthanizing rats was tested for vitamin-B12. PCs in stomach fundus were examined using histological and transmission electron microscopic studies, besides immunohistochemistry for Caspase-3, proliferating cell nuclear antigen (PCNA) and hydrogen potassium (H+/K+)-ATPase. Gastric homogenates were inspected for malondialdehyde (MDA) by ELISA. TAM intake induced structural and ultrastructural alteration in rat PCs including ballooning degeneration, apoptosis, decreased canaliculi, increased tubulovesicular system and irregular-shaped mitochondria. A significant increase of Caspase-3 immunostaining and MDA expression in gastric tissue was associated with a significant decrease of PCNA and H+/K+-ATPase-immunostaining and in serum vitamin-B12 as compared to the control group. Combined oral intake of TAM and Rh-WE revealed a significant reversal of the previous findings. Conclusion: Prolonged use of oral TAM substantially affected the structure and function of gastric PCs which can be ameliorated by Rh-WE.

Catechins promoted <i>Enterococcus faecalis</i> to alleviate related indices of nonalcoholic steatohepatitis mice induced by high-fat diet

IntroductionWild Cordyceps sinensis (C. sinensis) is a Chinese medicinal material known for its renal and pulmonary benefits. However, inorganic arsenic in wild Cordyceps sinensis may accumulate in the body following prolonged consumption; therefore, rigorous safety evaluations are needed.MethodsThis study compared the impacts of wild Cordyceps sinensis at the maximum clinical dose and equivalent doses of inorganic arsenic (16.36 mg/kg) to its total arsenic dose on organ indices, arsenic accumulation, and functional and pathological changes in the liver and kidney in mice, aiming to explore the safety of consuming wild Cordyceps sinensis. Arsenic accumulation in organs was measured via ICP–MS, while serum markers of liver and kidney functions were assessed via ELISA and biochemical assay kits. Histopathology was observed through H&E staining.ResultCompared with those in the control group, no significant adverse effects on body weight, organ indices, arsenic accumulation, liver or kidney function, or liver or kidney pathology were observed in the Cordyceps group. In contrast, inorganic arsenic exposure resulted in significant arsenic accumulation in organs, especially in the liver and kidneys (p < 0.01), liver and kidney function impairment (p < 0.01), and pathological changes, including hepatic steatosis, mild edema, balloon degeneration, and renal tubular epithelial cell edema and degeneration, with the aggregation of eosinophils in the renal capsule.ConclusionThese findings indicate that, at the maximum clinical dose, wild Cordyceps sinensis does not cause measurable hepatorenal toxicity in long-term and exhibits markedly greater safety compared to a mixture of inorganic arsenic compounds (sodium arsenate and sodium arsenite) at an equivalent total arsenic dose.

AimsTo reveal clinicopathological characteristics of alcoholic foamy degeneration (AFD)―an uncommon form of alcoholic liver injury.MethodsClinicopathological features of AFD (n=9) were examined in comparison to those of severe alcoholic hepatitis (SAH;...

Indian childhood cirrhosis is a chronic liver disease in infants and children. Indian childhood cirrhosis is unique to the Indian subcontinent and occurs from 6 months to 5 years of age. We report 2 cases in a period of 5 years, including 1 male and 1 female. Both children were less than 3 years of age. Presenting complaints were jaundice and hepatosplenomegaly. The clinical diagnosis was metabolic liver disease. Histological findings included diffuse hepatocellular ballooning degeneration, prominent Mallory Denk bodies, diffuse pericellular fibrosis, and marked copper/copper-associated protein deposits, along with the absence of steatosis and glycogenated nuclei. Mettalothionein immunohistochemistry was performed in 1 case and showed strong positivity. The first child developed liver failure and died. The second child was started on oral penicillamine therapy and is alive on the most recent follow-up. Whole-exome studies of both patients showed no significant findings. None of the children had exposure to excess dietary copper. Sporadic cases of Indian childhood cirrhosis continue to occur. There should be greater awareness among pediatricians and pathologists of the disease to enable earlier diagnosis. Awareness of metallothionein expression in biopsies of patients with Indian childhood cirrhosis is important to prevent misdiagnosis of Wilson disease.

Outbreak of ruminal acidosis in cattle caused by the ingestion of hedge lucerne (Desmanthus virgatus L. Willd.) in Northeastern Brazil

A temporal relationship between liver enzyme derangement and an herbal remedy warrants further assessment for herb‐induced liver injury (HILI). Here, we describe the use of kava, a drink traditionally consumed in Pacific Island cultures, causing acute ALT and AST elevation as assessed by an updated RUCAM score of 7. The increasing use of kava in Western society should prompt clinicians to be more aware of this rare cause of HILI. A 46‐year‐old man was referred to the emergency department with a 3‐week history of fatigue, right upper quadrant pain, and profound transaminitis. He commenced kava 10 g daily 5 weeks prior to aid sleep, which was ceased 2 weeks prior due to his biochemical derangement. Blood tests revealed an ALT of 1546 U/L and an AST of 920 U/L. An autoimmune screen, viral serology, and liver ultrasound showed no abnormalities. A liver biopsy revealed foci of hepatocellular necrosis with scattered ballooning degeneration and apoptotic bodies in the parenchyma, but normal underlying hepatic parenchyma without steatosis. Following cessation of kava, the liver enzymes improved without any other intervention. He was monitored as an outpatient and had no recurrence. The incidence of kava HILI may increase with its marketing; its exact mechanism is unknown. Ultimately, further research is needed to identify the pathogenesis of kava HILI. HILI is a significant cause of transaminitis, and clinicians should remain vigilant in patients presenting with nonspecific symptoms and a negative liver screen.

ABSTRACTBackgroundMpox (formerly known as monkeypox), a zoonotic disease caused by Monkeypox virus (MPXV), has become an international outbreak since May 2022. Mpox often presents with a mild systemic illness and a characteristic vesiculopustular skin eruption. In addition to molecular testing, histopathology of cutaneous lesions usually shows distinctive findings, such as epidermal necrosis, balloon degeneration, papillary dermal edema, and focal dermal necrosis, which have proven helpful in the diagnosis of mpox. Viral cytopathic changes with areas of multinucleation, smudging of the nuclei, and intracytoplasmic inclusions have also been described. Although useful, these features are relatively nonspecific. The use of a monoclonal antibody for immunohistochemical (IHC) staining of MPXV may be a useful tool in confirming mpox infection.MethodsThree cases of PCR‐confirmed mpox were biopsied and subjected to IHC staining with a monoclonal MPXV‐specific antibody targeting viral envelope protein A29. As controls, cell lines transduced to express other MPXV viral antigens and samples of cutaneous viral infections involving Molluscum contagiosum, Herpes simplex, Herpes zoster, or Cytomegalovirus were also subjected to IHC staining with this antibody.ResultsAll three mpox patient biopsies performed on lesional skin subjected to MPXV IHC staining reliably detected viral infection in lesional skin with a diffuse cytoplasmic and focally nuclear staining pattern. No staining was seen in transduced cell lines expressing off‐target MPXV viral antigens and in lesional skin of other common viral infections listed above.ConclusionsThe monoclonal MPXV‐specific antibody may be used as an adjunct tool to confirm mpox infection.

Gossypol is an anti-nutritional factor in cottonseed products, and its toxicity is reduced mainly by transforming free gossypol (FG) to bound gossypol (BG). However, the safety of BG chronic consumption in relation to liver and reproduction has not been fully examined. This study aimed to assess the effects of chronic BG exposure on liver and male reproductive function. BG with a purity of 97.38 % was prepared from whole egg solution and acetate gossypol, and its digestive stability was investigated by in vitro and in vivo digestion experiments. In vitro , digestion experiment of BG in the simulated gastric and intestinal fluid was performed. In vivo , thirty 8-week-old male C57BL/6 mice were randomly assigned into 3 groups, and the effects of BG on the liver and reproductive function of mice were studied by administering 24.8 mg/(kg·BW) BG (contains 0.67 mg FG) (marked as BG group), through gavage for 90 days, and administrated with 0.67 mg/(kg·BW) FG as a control group (C group), administrated with vehicle as a negative control group. Results show that BG dissociated into FG in simulated gastrointestinal environments. Long-term BG administration hindered the growth performance of the male mice, during which higher FG was detected in the feces of BG-administrated mice. This BG administration also increased serum alanine aminotransferase and aspartate aminotransferase levels, as well as the balloon degeneration and binucleate cells in the liver. Sperm concentration, sperm motility, epididymis coefficient, and the lactate dehydrogenase-X activity in the testis were inhibited by BG administration. Taken together, we reported that long-term administration with BG at a low dosage of 24.8 mg/(kg·BW) damages the growth performance, liver, and reproductive function, which may guide the modest use of cottonseed in foods and feeds.

Short-chain chlorinated paraffins induce liver injury in mice through mitochondrial disorders and disruption of cholesterol-bile acid pathway

Abstract Disclosure: C. Alvarado: None. L.K. Beitel: None. M. Paliouras: None. M.A. Trifiro: None. The mouse androgen receptor (AR) shares over 90% homology with the human ortholog, but it lacks the CAG encoded poly-Q tract; instead, it contains a mixed glutamine/histidine tract. To study AR functionality in a more structured fashion, we created a knock-in mouse model that has only the equivalent of human poly-Q tract while maintaining the remaining mouse receptor sequence intact. Our humanized androgen receptor mouse (AR-19Q) displays a phenotype that is best described as Metabolic Syndrome (MetS). The mice gain excess weight, have hypertension, hyperglycemia, pancreatic islet cell abnormalities, and progressive non-alcoholic fatty liver disease (NAFLD). Features of MetS begin at approximately 4 months and progress till 12-14 months. Obesity: AR-19Q mice are significantly heavier, with a 35% gain of weight vs C57BL/6 mice. However, these mice are not hyperphagic at any time and eat identical quantities of chow. To assess obesity status, and found AR-19Q mice to have a significantly higher depositions of total white adipose visceral, lingual and subcutaneous vs age-matched controls. NAFLD: The progression of NAFLD is observed in AR-19Q mice. AR-19Q Histological analysis and immunohistochemistry reveal microvesicular steatosis, superseded by macrovesicular steatosis with hepatocyte balloon degeneration, scattered lobular inflammation (macrophages and lymphocytes) (6-9 months) and further superseded by perisinusoidal fibrosis (12 months). Biochemical blood analysis confirms. that there is a significant increase in alanine aminotransferase (ALT) levels, indicative of liver damage. With time classical NASH hepatitis and subsequent NASH fibrosis ensues; with several mice developing hepatocellular carcinoma (overall 20% of mice). Gene expression profiling AR-19Q mice for fatty liver genes, we observe an upregulation of inflammation genes Il-6, Il-10 and Tnf. In addition, increased Lpl (lipoprotein lipase) is indicative of increased blood triglyceride levels. On the other hand, down-regulation of G6pc (glucose-6-phosphatase) could be linked to glycogen storage disease. Down-regulation of Slc2a2 and Slc2a4, that encode GLUT2 and GLUT4, should have an influence on glucose levels, glucose homeostasis and insulin sensitivity, as insulin promotes glucose uptake through GLUT4. Type 2 Diabetes: Glucose intolerance is observed by 3-4 months followed by fasting hyperglycemia. Pancreases show advanced islet hyperplasia, concurrent with insulin resistance and type 2 diabetes. Pancreases of middle-age AR-19Q mice present with a significant islet hyperplasia, suggesting peripheral insulin resistance. We propose the use of the AR-19Q mice as a model to study the metabolic dysfunction/ deficiency leading to the MetS. The model will allow us to a better understanding into the mechanisms responsible for T2D and NAFLD-related disorders. Presentation: 6/2/2024

Abstract Disclosure: C. Alvarado: None. L.K. Beitel: None. M. Paliouras: None. M.A. Trifiro: None. The mouse androgen receptor (AR) shares over 90% homology with the human ortholog, but it lacks the CAG encoded poly-Q tract; instead, it contains a mixed glutamine/histidine tract. To study AR functionality in a more structured fashion, we created a knock-in mouse model that has only the equivalent of human poly-Q tract while maintaining the remaining mouse receptor sequence intact. Our humanized androgen receptor mouse (AR-19Q) displays a phenotype that is best described as Metabolic Syndrome (MetS). The mice gain excess weight, have hypertension, hyperglycemia, pancreatic islet cell abnormalities, and progressive non-alcoholic fatty liver disease (NAFLD). Features of MetS begin at approximately 4 months and progress till 12-14 months. Obesity: AR-19Q mice are significantly heavier, with a 35% gain of weight vs C57BL/6 mice. However, these mice are not hyperphagic at any time and eat identical quantities of chow. To assess obesity status, and found AR-19Q mice to have a significantly higher depositions of total white adipose visceral, lingual and subcutaneous vs age-matched controls. NAFLD: The progression of NAFLD is observed in AR-19Q mice. AR-19Q Histological analysis and immunohistochemistry reveal microvesicular steatosis, superseded by macrovesicular steatosis with hepatocyte balloon degeneration, scattered lobular inflammation (macrophages and lymphocytes) (6-9 months) and further superseded by perisinusoidal fibrosis (12 months). Biochemical blood analysis confirms. that there is a significant increase in alanine aminotransferase (ALT) levels, indicative of liver damage. With time classical NASH hepatitis and subsequent NASH fibrosis ensues; with several mice developing hepatocellular carcinoma (overall 20% of mice). Gene expression profiling AR-19Q mice for fatty liver genes, we observe an upregulation of inflammation genes Il-6, Il-10 and Tnf. In addition, increased Lpl (lipoprotein lipase) is indicative of increased blood triglyceride levels. On the other hand, down-regulation of G6pc (glucose-6-phosphatase) could be linked to glycogen storage disease. Down-regulation of Slc2a2 and Slc2a4, that encode GLUT2 and GLUT4, should have an influence on glucose levels, glucose homeostasis and insulin sensitivity, as insulin promotes glucose uptake through GLUT4. Type 2 Diabetes: Glucose intolerance is observed by 3-4 months followed by fasting hyperglycemia. Pancreases show advanced islet hyperplasia, concurrent with insulin resistance and type 2 diabetes. Pancreases of middle-age AR-19Q mice present with a significant islet hyperplasia, suggesting peripheral insulin resistance. We propose the use of the AR-19Q mice as a model to study the metabolic dysfunction/ deficiency leading to the MetS. The model will allow us to a better understanding into the mechanisms responsible for T2D and NAFLD-related disorders. Presentation: 6/2/2024

In China, there is a long history and rich clinical experience in treating nonalcoholic steatohepatitis (NASH) with traditional Chinese herbal medicines, including Chai Hu Shu Gan San. This study aims to investigate the potential regulatory effects of Chaihu Shugan San (CSS) on liver lipid metabolism and inflammatory damage in mice with experimental nonalcoholic steatohepatitis (NASH) induced by a choline-deficient high-fat diet (CDHFD). Utilizing network pharmacology, we systematically explore the mechanisms of action and therapeutic potential of CSS against NASH. Potential targets in CSS and targets for NASH were identified using online databases. Functional enrichment and protein-protein interaction analyses were conducted to identify hub-targeted genes and elucidate the underlying molecular mechanisms. The affinities of active compounds in CSS with hub-targeted genes were evaluated using molecular docking. Finally, hub-targeted genes were validated through real-time polymerase chain reaction, western blotting, and immunofluorescence in choline-deficient high-fat diet mice, both with and without CSS treatment. CSS reduces serum ALT and AST levels in NASH mice(P < 0.05) and ameliorates ballooning degeneration in the livers of NASH mice, thereby lowering the NAS score(P < 0.05). Including naringenin, high-performance liquid chromatography/mass spectrometrys identified 12 chromatographic peaks. Based on network pharmacology analysis, CSS contains a total of 103 active compounds and 877 target genes. Transferase activity represents a potential mechanism for therapeutic intervention of CSS in NASH. The transcriptional levels and protein expression of the SIRT1 gene in NASH mice are significantly increased by CSS (P < 0.05). Naringenin is probable active compound in CSS and SIRT1 is the hub gene by which CSS is involved in NASH treatment.

Obesity and its associated hepatic steatosis have become a global concern, posing numerous health hazards. Photodynamic therapy (PDT) is a unique approach that promotes anti-obesity by releasing intracellular fat. Chlorin e6 (Ce6)-PDT was tested for its anti-obesity properties in male ovariectomized (OVX) beagle dogs, as well as male C57BL/6 and Balb/c mice. The 12 OVX beagles were randomly assigned to one of four groups: high-fat diet (HFD) only, Ce6 only, Ce6 + 10 min of light-emitting diode light (LED) treatment, and Ce6 + 15 min of light treatment. We assessed several parameters, such as body weight, adipose tissue morphology, serum biochemistry, and body fat content analysis by computed tomography (CT) scan in HFD-fed beagle dogs. At the end of the study period, dogs that were treated for 35 days with Ce6 and exposed to LED irradiation (660 nm) either for 10 min (Ce6 + 10 min of light) or for 15 min (Ce6 + 15 min of light) had decreased body weight, including visceral and subcutaneous fats, lower aspartate transaminase (AST)/alanine transaminase (ALT) ratios, and a reduction in the area of individual adipocytes with a concomitant increase in the number of adipocytes. Furthermore, C57BL/6 male mice following an HFD diet were effectively treated by Ce6-PDT treatment through a reduction in weight gain and fat accumulation. Meanwhile, Ce6-PDT attenuated hepatocyte steatosis by decreasing the epididymal adipose tissue and balloon degeneration in hepatocytes in HFD-fed Balb/c mice. Taken together, our results support the idea that Ce6-PDT is a promising therapeutic strategy for the recovery of obesity and obesity-related hepatic steatosis.

SQUIRRELPOX has been identified as a key factor in red squirrel (Sciurus vulgaris) decline in the UK.1 Grey squirrels (Sciurus carolinensis) are thought to act as reservoir hosts for squirrelpox virus (SQPV), the causative agent of squirrelpox, with a reported asymptomatic seroprevalence of 61 per cent.2SQPV is implicated in the complete replacement of red squirrels by grey squirrels throughout mainland England and Wales,1 and poses a major threat to Scottish red squirrel populations since being first detected in 2007.3 To combat this threat, an mortality surveillance programme has been established at the Royal (Dick) School of Veterinary Studies, University of Edinburgh, using opportunistic sampling of red squirrel carcases. This has been ongoing for several years, including a summary publication covering 262 cases submitted from 2005 to 2009.4As part of this monitoring programme, an adult female red squirrel carcase was submitted by a member of the public, after having been found in Townhall Wood (NT110894), on the outskirts of Dunfermline, Fife, in March 2024. Postmortem examination identified multiple lesions typically associated with squirrelpox, including ulcerative and exudative dermatitis of the periocular and perioral skin (Fig 1). Histopathology of the affected skin identified extensive ulceration alongside remnant areas of epithelium with marked ballooning degeneration and eosinophilic intracytoplasmic inclusion bodies. Transmission electron microscopy of the affected tissue also identified numerous pox virions within the affected tissue, which through their size, shape and available surface morphology (Fig 2), were consistent with those of SQPV. In Britain there have been no additional identified diseases in red squirrels that present with periocular or perioral, ulcerative to exudative dermatitis due to a poxvirus,5indicating this case is highly likely due to SQPV.This finding represents the first identification of squirrelpox north of the central belt and is consistent with the predictions of previous modelling, which identified a high risk of northern SQPV spread from 2023 onwards.6 This modelling also suggests a rapid increase and spread of squirrelpox into more northerly and naive red squirrel populations is likely following establishment north of the central belt in central Scotland.6This case and modelling supports an increased requirement for targeted investigations, ongoing monitoring and grey squirrel interventions both around Dunfermline itself and within adjacent areas to establish the disease burden in this locality and limit further northerly squirrelpox spread.LA Wilson, veterinary pathology lecturer, M Marr, postdoctoral research fellow, C Logie, postmortem room technician, K Beckmann, conservation medicine lecturer, PWW Lurz, squirrel ecologist, R Ogden, director of conservation science, E Milne, professor emerita of veterinary clinical pathology Royal (Dick) School of Veterinary Studies and Roslin Institute, University of Edinburgh, Easter Bush Campus, Roslin, Midlothian EH25 9RG email: liam.wilson@ed.ac.uk DJ Everest, pathology scientist APHA Weybridge, Addlestone, Surrey KT15 3NBReferences1 Tompkins DM, White AR, Boots M. Ecological replacement of native red squirrels by invasive greys driven by disease. Ecol Lett2003;6:189–962 Sainsbury AW, Nettleton P, Gilray J, et al. Grey squirrels have high seroprevalence to a parapoxvirus associated with deaths in red squirrels. Anim Conserv2000;3:229–33 3 Mclnnes CJ, Coulter L, Dagleish MP, et al. First cases of squirrelpox in red squirrels (Sciurus vulgaris) in Scotland. Vet Rec2009;164:528–314 LaRose JP, Meredith AL, Everest DJ, et al. Epidemiological and postmortem findings in 262 red squirrels (Sciurus vulgaris) in Scotland, 2005 to 2009. Vet Rec2010;167:297–3025 Everest DJ, Tolhurst-Cherriman DAR, Davies H, et al. Assessing a potential non-invasive method for viral diagnostic purposes in European squirrels. Hystrix 2019;30:44–506 White A, Lurz PWW. A modelling assessment of control strategies to prevent/reduce squirrelpox spread. 2014. Scottish Natural Heritage Commissioned Report No 627. https://bit.ly/441TOgM (accessed 8 April 2024)PROFESSIONHistory of the veterinary professionI WRITE in response to the debate article by Bruce Vivash Jones (VR, 16/23 March 2024, vol 194, p 236). As a PhD researcher on the history of the RCVS and veterinary regulation, and a veterinary nurse, I would like to raise some issues with Vivash Jones’ historical evidence. He states that changes to the council structure would create an oligarchy. From his interpretation of an oligarchy I can assure him and our profession that the first RCVS council did work as This finding represents the first identification of squirrelpox north of the central belt20/27 April 2024 | VET RECORD312Fig 2: Squirrelpox virus virions detected in the affected tissue. Bar = 200 nmFig 1: Macroscopic lesions of ulcerative and exudative dermatitis surrounding the eye in a red squirrel (Sciurus vulgaris)Letters 20 April.indd 312Letters 20 April.indd 31216/04/2024 12:4816/04/2024 12:48