In vivo最新文献

Background/aim: Metabolic dysfunction-associated steatotic liver disease (MASLD), formerly known as nonalcoholic fatty liver disease (NAFLD), is a significant global health burden, with increasing evidence pointing to gut microbiota as a modifiable contributor to disease pathogenesis. Yet, the causality of gut microbiota in liver health is not fully established. This study aimed to clarify a causal relationship of specific gut microbes with MASLD.

Materials and methods: In this pre-clinical study, we assessed the hepatoprotective potential of Bacteroides eggerthii (B. eggerthii), a gut commensal bacterium, in a high-fat diet (HFD)-induced mouse model of MASLD.

Results: Our in vivo experiments showed that mice given with HFD exhibited prominent metabolic abnormalities, intestinal dysbiosis, liver steatosis, and drivers of pathological changes in the livers (autophagic dysregulation and lipid peroxidation). Oral supplementation with a strain of B. eggerthii that was isolated from the fecal specimen of a healthy subject improved the perturbation of fecal microbiota and mitigated lipid peroxidation and lobular inflammation in the liver of MASLD mice, without ameliorating hyperlipidemic conditions and hepatic steatosis.

Conclusion: B. eggerthii may potentially protect against diet-induced fatty liver diseases and further extend our understanding on the gut-liver axis in MASLD.

Background/aim: Induction of immune tolerance by the activation of regulatory T (Tregs) cells after extracorporeal photopheresis (ECP) appears to influence the response to treatment in patients with chronic graft-versus-host disease (cGvHD). This study examined the activation mechanisms of Tregs and their potential link to the expression of FOXP3, TP53, and SELPLG genes. Results were compared with the therapeutic response to ECP.

Materials and methods: The study included six patients with cGvHD who underwent at least four cycles of ECP. The samples were collected from peripheral blood (PB) and from apheresis material before and after ECP, which was then used to establish cell cultures. The percentage of Tregs and their subpopulations were assessed using multicolor flow cytometry. Gene expression levels were evaluated using quantitative PCR (qPCR).

Results: A statistically significant increase in the expression levels of TP53 gene was found in the fourth ECP cycle in patients with partial response (PR) compared to patients with stable disease (SD). Patients with PR were characterized by higher mRNA levels for the TP53 and SELPLG genes after 48 h of culture. Patients with PR had a statistically significant higher percentage of Tregs including activated HLA-DR+ Tregs.

Conclusion: Patients with PR demonstrated elevated gene expression levels in 48-h mononuclear cell cultures, which correlated with their clinical response to treatment. They also had a higher baseline percentage of Tregs, which did not correlate with elevated FOXP3 or other gene expression levels.

Background/aim: Although the purse-string suture (PSS) method is widely adopted to reduce surgical-site infection (SSI; the commonest complication following bowel-stoma closure), this technique is associated with delayed epithelialization. Recently, delayed primary closure (DPC) combined with negative-pressure wound therapy (NPWT) has been proposed as a promising alternative. This study aimed to evaluate the clinical efficacy of DPC with NPWT compared to the PSS method in stoma-closure surgery.

Patients and methods: We conducted a retrospective observational study involving 31 patients who underwent stoma closure between January 2021 and March 2023. Patients were categorized into two groups: PSS with or without NPWT (PSS±NPWT; n=15) and DPC with NPWT (DPC+NPWT; n=16). The primary outcome was wound-healing duration; the secondary outcomes were patient satisfaction and in-hospital costs.

Results: The wound-healing duration was significantly shorter in the DPC+NPWT group compared to the PSS±NPWT group (median: 20 vs. 45 days, p<0.001). Multivariate analysis identified wound-closure method as an independent predictor of wound healing at 30 days. Patient-reported outcomes indicated improved wound-care experience and a trend toward better cosmetic satisfaction in the DPC+NPWT group. Although not statistically significant, the total in-hospital costs were lower in the DPC+NPWT group despite higher device-related expenditures.

Conclusion: DPC with NPWT significantly shortened the wound-healing duration and showed potential benefits regarding patient satisfaction and healthcare costs compared to the PSS method. These findings suggest DPC with NPWT as a viable alternative for bowel-stoma closure and wound management.

The sural nerve grafting plays a crucial role in the repair of peripheral nerve impairments when tension-free neurorrhaphy is unfeasible. The sural nerve is frequently utilized in sensory and mixed nerve repairs because of its anatomical reliability, sensory characteristics, and low donor site morbidity. This article critically reviews the current knowledge in the field and examines the anatomical and histological features of the sural nerve, along with contemporary harvesting techniques and clinical applications. High-resolution ultrasound (US) is highlighted as a non-invasive, real-time instrument for preoperative preparation, intraoperative mapping, and postoperative assessment. Ultrasonography enhances procedural precision, resulting in superior outcomes and less complications. Given the constraints of sural nerve grafts, alternative solutions such processed allografts and synthetic guidance are being evaluated.

Background/aim: Manual syringe injections often produce variable pain responses due to inconsistent injection pressure and speed, complicating both clinical procedures and experimental outcomes. The digital automatic syringe, I-ject™, was developed to provide controlled, reproducible injections and thereby reduce injection-associated pain.

Materials and methods: Using a formalin-induced acute pain model in rats, we compared nociceptive behavioral and molecular responses between injections delivered by a conventional manual syringe versus the I-ject™ automatic syringe. Pain-related behaviors, limping and licking of the injected paw, were recorded for 60 min. After behavioral observation, spinal cord tissue was collected for quantitative real-time PCR analysis of pain-related genes.

Results: Rats injected with the I-ject™ automatic syringe exhibited a 30.6% reduction in limping and a 66.0% reduction in licking frequency within the 15 min after formalin injection, compared to those injected with a manual syringe (p<0.05). Expression of examined pain-marker genes was significantly lower in the automatic syringe group than in the manual syringe group. Notably, c-fos and GFAP mRNA levels were reduced by 29.7% and 81.8%, respectively. Similar downregulation trends were observed for Oprl1, Htr2a, and Oprm1, indicating that the automatic injection attenuated both neuronal and glial activation associated with pain.

Conclusion: The I-ject™ automatic syringe markedly reduced both behavioral signs of pain and molecular markers of nociceptive activation in this rodent formalin pain model. These findings support the potential of this digital automatic injection system as a safer and more consistent alternative to conventional manual syringes in both preclinical research and clinical settings.

Background/aim: Aging results in diminished physiological functions and reduced stem cell activity, driving research into cellular mechanisms and natural compounds that support tissue regeneration and improve skin health. Marine algae, particularly Caulerpa lentillifera (sea grape), are rich in sulfated polysaccharides and widely used in the food and cosmetics industries due to their diverse biological properties, including UV protection, anti-inflammatory effects, and hydration.

Materials and methods: The expression levels of stem cell transcription factors were evaluated at the mRNA level using RT-qPCR and at the protein level via western blot and immunofluorescence. Melanin content was quantified in human melanoma cells treated with sea grape extract.

Results: The findings revealed that treatment with sea grape significantly enhanced the expression of stem cell transcription factors in human Dermal Papilla (DP) cells, with mRNA levels of OCT4, NANOG, and SOX2 increasing by 20-, 35-, and 15-fold, respectively. Likewise, in UE6E7T-3 bone marrow stem cells, sea grape induced OCT4, NANOG, and SOX2 induction by 1.5-, 1.5-, and 2.5-fold, respectively. Corresponding protein levels of these stem cell markers also showed increased expression in both sea grape-treated DP cells and UE6E7T-3 cells. Additionally, sea grape exhibited a significant anti-melanogenic effect on human melanoma cells.

Conclusion: Overall, sea grape reduces melanin levels and stimulates regenerative markers in stem cells, presenting a promising opportunity for developing novel products focused on pigmentation regulation and skin rejuvenation.

Background/aim: Complement activation has been implicated as a contributor to lung injury. Membrane-bound complement regulatory proteins, including decay-accelerating factor (DAF), control complement activation, thus mitigating complement-mediated injury. Their effect on lung injury remains unexplored. Using a DAF knockout rat model, we assessed the effect of DAF on the extent of lipopolysaccharide (LPS)-mediated acute lung injury (ALI) in rats.

Materials and methods: Wild-type (WT) and DAF-knock out (Daf^-/- ) rats were injected intraperitoneally with a sublethal LPS dose. Following 16 h, protein and cytokine levels were assessed using immunohistochemistry/immunoblotting and ELISA, respectively.

Results: LPS administration to WT rats caused histopathological lesions indicative of ALI and significantly increased infiltrating inflammatory cells, as well as total number of cells and interleukin (IL)-6 levels, in bronchoalveolar lavage fluid samples. DAF absence resulted in increased C3b levels in Daf^-/- control rats, compared with their WT controls. LPS administration to Daf^-/- rats exacerbated lung injury without an effect on C3b levels. Higher baseline tissue protein levels of the anaphylatoxin complement component 5a receptor 1 (C5aR1) were observed in Daf^-/- rats compared with WT whereas LPS administration to either group resulted in reduced levels of C5aR1, albeit not significant. DAF deficiency had no effect on baseline protein levels of CR1-related gene/protein Y (Crry) complement regulator, whereas LPS administration increased Crry protein levels to a similar extent in WT and Daf^-/- rats.

Conclusion: DAF deficiency increased baseline C3b and C5aR1 protein levels in lung tissue. LPS-induced inflammatory response and lung injury were augmented in Daf^-/- rats without further increase in C3b and in the absence of significant changes in the expression of membrane-bound complement regulatory proteins, DAF and Crry, or the anaphylatoxin receptor C5aR1.

Background/aim: The adverse side effects associated with chemotherapeutic agents have prompted the exploration of natural compounds as adjuvants to chemotherapy, offering more effective therapeutic alternatives. Licoricidin, a bioactive constituent of licorice, possesses diverse pharmacological properties. However, the antitumor mechanisms underlying the therapeutic effects of Licoricidin combined with paclitaxel in cervical cancer remain unclear and further investigations are warranted.

Materials and methods: Cell growth and apoptosis were assessed using the MTT assay and Annexin V/PI staining. Endoplasmic reticulum (ER) stress was evaluated using the ER-ID assay. The expression of apoptosis- and ER stress-related proteins in response to the combined treatment was analyzed using western blotting.

Results: Combined treatment with licoricidin and paclitaxel effectively inhibited cell growth and induced apoptosis in human HeLa and C-33A cells. We further revealed that this combined treatment increased the expression of apoptotic proteins (c-PARP, Bax, and DR5) while decreasing the expression of pro-caspase proteins (pro-caspase-3 and pro-caspase-9) in HeLa cells. In addition, the combined treatment induced ER stress, as evidenced by decreased expression levels of ERp44, ERp57, and ERp72, whereas the expression of GRP78 was increased in HeLa cells.

Conclusion: The combination of licoricidin and paclitaxel induced ER stress-mediated apoptosis and may serve as a potential antitumor therapeutic strategy against human cervical cancer cells.

Background/aim: The optimal extent of neck dissection (ND) in oral squamous cell carcinoma (OSCC) is controversial, particularly regarding levels IV and V in cases with metastases in levels I-III. This study evaluated the probability of metastases in levels IV-V when levels I-III are pN+ in cN0 necks and analyzes prognostic factors influencing their occurrence.

Patients and methods: A retrospective study was performed at the Department of Oral, Maxillofacial and Facial Plastic Surgery, Ludwigshafen Hospital, Germany, including 61 patients with primary OSCC treated surgically including ND. Patients underwent either supraomohyoid ND (SOND) of levels I-III with secondary extension to IV-V or modified radical ND (MRND) of levels I-V. Statistical analysis assessed the correlation between metastases in levels IV-V and extracapsular spread (ECS), number of positive lymph nodes, T-classification, bone infiltration, grading, lymphovascular invasion, vascular invasion, and perineural invasion.

Results: Among the 61 patients with metastases in levels I-III, 6 patients (9.8%) had metastases in levels IV-V. A significant correlation (p=0.042) indicated that pN+ in levels I-III is associated with >5% risk of level IV-V metastases. The presence of more than one metastasis in levels I-III significantly (p=0.027) predicted level IV-V involvement. A pN status of >pN2b significantly (p=0.002) increased the prevalence of metastases in levels IV-V. ECS showed a trend toward increased IV-V involvement, though not statistically significant (p=0.078).

Conclusion: The risk of level IV-V metastases in patients with pN+ in levels I-III exceeds 5% in cN0 necks. The number of affected nodes and pN classification were the strongest predictors. These findings support selective extension of ND beyond level III in specific patients and emphasize individualized treatment strategies.

Background/aim: To compare survival outcomes and radiation-related toxicities between conventional radiotherapy (CvT) and volumetric-modulated arc therapy (VMAT) in patients with early glottic cancer.

Patients and methods: We reviewed 81 patients with Tis-T2N0 glottic cancer who underwent definitive radiotherapy between 2010 and 2018. CvT (N=47) was delivered using two opposing lateral beams (70 Gy in 35 fractions), while VMAT (N=34) utilized two arcs (65.25 Gy in 29 fractions). Hypothyroidism was defined as an elevated thyroid-stimulating hormone level with or without a decrease in free T4 or T3 levels.

Results: The overall median follow-up time was 80.0 months and was longer in the CvT group (116.0 months) than in the VMAT group (45.3 months). Five-year local failure-free survival (95.4% vs. 84.5%, p=0.088), regional failure-free survival (93.0% vs. 94.1%, p=0.481), locoregional failure-free survival (90.9% vs. 84.7%, p=0.319), and overall survival (89.4% vs. 90.7%, p=0.575) were comparable between the CvT and VMAT groups, respectively. No increase in regional failure was observed in the VMAT group despite exclusion of the anterior level III neck. Most local failures in the VMAT group occurred near the original tumor site and were associated with T2 disease (p=0.015). Hypothyroidism occurred more frequently in the CvT group (53.5%) than in the VMAT group (25.9%, p=0.043). Carotid artery events were noted in three patients treated with CvT (6.4%), with none in the VMAT group.

Conclusion: VMAT demonstrated comparable survival outcomes to CvT while significantly reducing the incidence of hypothyroidism and potentially lowering carotid artery events. VMAT may be considered a preferred modality to improve quality of life in patients with early glottic cancer.