Fusobacterium nucleatum facilitates anti-PD-1 therapy in microsatellite stable colorectal cancer.

Microsatellite stable (MSS) colorectal cancers (CRCs) are often resistant to anti-programmed death-1 (PD-1) therapy. Here, we show that a CRC pathogen, Fusobacterium nucleatum (Fn), paradoxically sensitizes MSS CRC to anti-PD-1. Fecal microbiota transplantation (FMT) from patients with Fn-high MSS CRC to germ-free mice bearing MSS CRC confers sensitivity to anti-PD-1 compared to FMT from Fn-low counterparts. Single Fn administration also potentiates anti-PD-1 efficacy in murine allografts and CD34+-humanized mice bearing MSS CRC. Mechanistically, we demonstrate that intratumoral Fn generates abundant butyric acid, which inhibits histone deacetylase (HDAC) 3/8 in CD8+ T cells, inducing Tbx21 promoter H3K27 acetylation and expression. TBX21 transcriptionally represses PD-1, alleviating CD8+ T cell exhaustion and promoting effector function. Supporting this notion, knockout of a butyric acid-producing gene in Fn abolishes its anti-PD-1 boosting effect. In patients with MSS CRC, high intratumoral Fn predicts favorable response to anti-PD-1 therapy, indicating Fn as a potential biomarker of immunotherapy response in MSS CRC.

The causal role between circulating immune cells and diabetic nephropathy: a bidirectional Mendelian randomization with mediating insights.

Diabetic nephropathy (DN) is a critical inflammatory condition linked to diabetes, affecting millions worldwide. This study employs Mendelian randomization (MR) to explore the causal relationship between immune cell signatures and DN, analyzing over 731 immune signatures and incorporating data from 1400 metabolites to investigate potential mediators. Despite no statistically significant influence of DN on immunophenotypes after FDR correction, some phenotypes with unadjusted low P-values warranted mention, including CD34 on Hematopoietic Stem Cell (Myeloid cell Panel), CD45 on CD33- HLA DR- (Myeloid cell Panel). Furthermore, three immunophenotypes were identified to have a significant impact on DN risk: CD16-CD56 on HLA DR+ NK (TBNK Panel), CD45 on HLA DR+ T cell (TBNK Panel), and CD33dim HLA DR+ CD11b+ AC (Myeloid cell Panel). Our findings underscore the critical role of immune cells in DN, highlighting potential mediators and offering new insights into its underlying mechanisms.

Immune checkpoint inhibitors and acute kidney injury.

As a new type of anti-tumor immunotherapy, immune checkpoint inhibitors (ICIs) have improved the prognosis of multiple malignancies. However, renal complications are becoming more frequent. Nephrotoxicity often manifests as acute kidney injury (AKI), and the most common histopathological type is acute tubulointerstitial nephritis (ATIN). Based on previous studies of the incidence and potential risk factors for nephrotoxicity, in this review, we describe the mechanism of AKI after ICIs treatment, summarize the incidence, risk factors, and outcomes of AKI, and discuss the diagnosis and management of immune checkpoint inhibitors-associated acute kidney injury (ICI-AKI). In addition, we review the current status of ICIs rechallenge and the therapeutic strategies of ICIs applied in kidney transplant recipients. Finally, we emphasize the importance of collaboration between nephrologists and oncologists to guide the treatment of ICIs and the management of renal complications.

Case report: Left atrial myxoma with morphology of cavernous hemangioma supplied by the right coronary artery.

Here, we report an unusual case of left atrial myxoma presented with morphology of cavernous hemangioma supplied by the right coronary artery. Surgical resection of the left atrium myxoma was performed, and the patient experienced an uneventful recovery during hospitalization.

Effect of BCG HSP70 Gene Transfection on Dendritic Cells Derived From Bone Marrow in Children With Acute Leukemia.

OBJECTIVES: In this study, immature dendritic cells (imDCs) were transfected with the Bacillé Calmette-Guérin (BCG) heat shock protein 70 (HSP70) gene to investigate the impact on the maturity and function of imDCs from the bone marrow of pediatric patients with acute leukemia. MATERIALS AND METHODS: Bone marrow mononuclear cells were isolated from pediatric patients with acute lymphoblastic leukemia who had achieved complete remission at least 6 months prior. The recombinant vector pDisplay-HSP70 was transfected into imDCs. The test groups included 5 subgroups: imDCs (imDCs without special processing), imDC-neos (imDCs transfected with the pDisplay vector), HSP70 (imDCs transfected with the pDisplay-HSP70 vector), tumor necrosis factor α (TNF-α) (imDCs induced with rhTNF-α), and HSP70+TNF-α. Mature dendritic cells (mDCs) from different groups (HSP70, TNF-α, and HSP70+TNF-α) and T cells were cultured. An equal number of lymphocytes and mDCs were used as controls. The proliferation indices of T cells and the cytokine contents (interleukin-12 and interferon-γ) were determined. RESULTS: The HSP70 group and the TNF-α group expressed higher levels of HLA-DR, CD80, and CD86 but lower levels than the HSP70+TNF-α group; there was no significant difference between the HSP70 group and the TNF-α group. The combination of HSP70 and TNF-α induced the highest levels of interleukin-12 and interferon-γ. CONCLUSIONS: The outcomes of this study indicated that gene transfection with BCG HSP70 evidently promoted imDC maturity and the antitumor effects of mDC-mediated T cells. It could serve as a candidate gene-modified cell vaccine for tumor immunotherapy.

[Determination of thiram, propineb and metiram in mushroom by gas chromatography-mass spectrometry].

OBJECTIVE: To establish a method for determination of thiram, propineb and metiram in mushroom samples by gas chromatography-mass spectrometry(GC-MS). METHODS: Insoluble heavy metal salts were converted into water-soluble sodium salts by alkaline buffer with strong chelating agents. Dithiocarbamates can be converted into different methyl ester compounds with ion pair methylation. The GC separation was performed on a DB-5 MS capillary column(30 m×0. 25 mm, 0. 25 µm). The pesticides were detected by GC-MS with selective ion monitoring(SIM) and quantified by external standard of working curve method. Methodsological verification was carried out based on optimized sample pretreatment and GC-MS condition. RESULTS: The concentrations of dithiocarbamates exhibited a good linear relationship with GC-MS within a certain range. The limits of detection of thiram, propineb and metiram were 0. 01, 0. 05 and 0. 05 mg/kg, respectively. Furthermore, the average recoveries were from 76. 98% to 93. 52%, and the maximum relative standard deviation was 11. 54%(n=6). CONCLUSION: This method is simple, sensitive, accurate and reliable. All the indices meet the requirements of pesticide residue detection.

Hypoxia-Induced Cleavage Of Soluble ephrinA1 From Cancer Cells Is Mediated By MMP-2 And Associates With Angiogenesis In Oral Squamous Cell Carcinoma.

INTRODUCTION: ephrinA1 plays important roles in tumor angiogenesis. Matrix metalloproteases (MMPs) can cleave ephrinA1 from the cell membrane into extracellular environment. However, how soluble ephrinA1 is modulated by hypoxia and whether MMPs participate in this hypoxic process remains to be investigated in detail. METHODS: Thirty-seven patients with oral squamous cell carcinoma (OSCC) were included in the present study for HIF-1α, MMP-2, MMP-9 and ephrinA1 detection by immunohistochemistry. Serum samples from 35 patients were collected both preoperatively and postoperatively to confirm the existence of soluble ephrinA1 by ELISA. Block assay and Western blot analysis were further carried out to elucidate the proteolysis mechanism of ephrinA1 under hypoxic condition in vitro. RESULTS: Our data demonstrated that HIF-1α, MMP-2, MMP-9 and ephrinA1 expressed positively, and correlated with microvessel density in OSCCs, except for MMP-9. The serum expression level of ephrinA1 in OSCC patients decreased significantly after surgical removal of the solid tumors. In vitro experiments indicated that GM6001, a MMP-specific inhibitor, could reduce hypoxia-induced soluble ephrinA1 secretion from SCC cells. Further Western blot analysis confirmed that both HIF-1α and MMP-2 were up-regulated by hypoxia in a similar time-dependent manner, with the MMP-9 expression unchanged during this course. CONCLUSION: These results suggested a possible novel mechanism that ephrinA1 secretion is mediated by HIF-1α/MMP-2 signaling cascade which may play pivotal roles in OSCC neovascularization in a paracrine manner.

[The study of combined unilateral intraoral and extraoral reduction approach in the treatment of anterior temporo-mandibular joint dislocation].

OBJECTIVE: To observe the clinical outcomes of a combined unilateral intraoral and extraoral reduction approach in the treatment of anterior temporomandibular joint (TMJ) dislocation. METHODS: Postural muscular chains were utilized in the biomechanical analysis of stomatognathic systems for improving TMJ repositioning approaches. A total of 87 patients with anterior TMJ dislocation were included in the present study. A combined unilateral intraoral and extraoral reduction approach was applied, and the clinical effects were evaluated. RESULTS: Biomechanical analysis reveal that reflexive contrac-tion of the maxillary muscle group was blocked sufficiently during the combined unilateral intraoral and extraoral reduction process. All dislocated TMJs were set successfully and efficiently with few complications. CONCLUSIONS: Combined unilateral intraoral and extraoral reduction approach is an effective, convenient, and minimally invasive way to treat anterior TMJ dislo-cations.

Growth arrest-specific protein 6 (Gas6) as a noninvasive biomarker for early detection of diabetic nephropathy.

BACKGROUND: To investigate the diagnostic level of cystatin C and growth arrest-specific gene 6 (Gas6) levels in elderly type 2 diabetic patients with different degrees of diabetic nephropathy (DN). METHODS: Four hundred and eighty-two old people, including 130 healthy controls, 130 normoalbuminuric diabetic patients, 122 with microalbuminuria, and 100 with macroalbuminuria, were recruited. Plasma Gas6 and serum cystatin C levels were measured. RESULTS: Plasma Gas6 concentration was significantly lower in diabetic patients with microalbuminuria or macroalbuminuria, as compared with diabetic subjects with normoalbuminuria; while cystatin C was significantly higher. Gas6 was inversely correlated with BMI, WHR, and HbA1c, while cystatin C was inversely correlated with urea nitrogen and creatinine. Multivariate logistic regression analysis showed that, after adjusted for established diabetes risk factors, higher plasma Gas6 was significantly associated with a decreased risk of DN, while higher serum cystatin C was significantly associated with an increased risk. Receiver operating characteristic curve analysis showed that Gas6 was better than cystatin C as a biomarker for early diagnosis and detection of DN, with a cutoff value of 9.435 ng/mL (86.1% sensitivity and 84.6% specificity). CONCLUSION: Compared to cystatin C, Gas6 may be potentially a better noninvasive diagnostic biomarker for early detection of DN.

Molecular characterization and expression of interleukin-10 and interleukin-22 in golden pompano (Trachinotus ovatus) in response to Streptococcus agalactiae stimulus.

In the present study, members of the interleukin (IL)-10 family of cytokines, including IL-10 (TOIL-10) and IL-22 (TOIL-22) of golden pompano (Trachinotus ovatus), were cloned for the first time, and their expression patterns and 3D structures analyzed. The full-length cDNA sequences of TOIL-10 and TOIL-22 contained open reading frames of 564 and 567 bp, respectively. TOIL-10 and TOIL-22 shared higher homology (78%-89%) with the corresponding genes from various fish relative to other species (25%-34%) and contained the IL-10 family signature and four cysteine residues that are well conserved in other vertebrate IL-10 members. Phylogenetic tree analysis of our sequences alongside other IL-10 family proteins revealed that TOIL-10 and TOIL-22 cluster together with other teleost IL-10 and IL-22 molecules. Expression of TOIL-10 and TOIL-22 genes was ubiquitous in all tissues examined. The TOIL-10 gene was also highly expressed in skin, heart, gill, spleen, kidney, brain and liver, and lower levels were detected in intestine and muscle. High expression of the TOIL-22 gene was observed in gill, intestine, kidney, spleen, with the lowest levels in liver. TOIL-10 and TOIL-22 were rapidly activated after SAΔphoB immunization and significantly increased to peak levels at 12 h and 4 d in golden pompano kidney and spleen respectively following challenge. Expression in the brain reached peak levels at 4 d and 3 d respectively after post-immunization. Our results collectively indicate that TOIL-10 and TOIL-22 participate in the host immune response to bacterial infection. Moreover, TOIL-22 plays a potentially important role in mucosal immunity.

A canine model of femoral head osteonecrosis induced by an ethanol injection navigated by a novel template.

There is no consensus on how to establish models of osteonecrosis of the femoral head (ONFH) in large mammals. The aim of this study was to investigate the effectiveness of a novel canine model of ONFH, induced by a navigated injection of absolute ethanol. Using three-dimensional reconstruction and rapid prototyping manufacturing techniques, a new template was designed and processed to navigate the ethanol injection. The femoral heads of 18 adult dogs were injected with ethanol. Macroscopic, X-ray and histological examinations were performed at 3, 6, and 9 weeks after the operation. Further, computed tomography (CT), magnetic resonance imaging (MRI), and radionuclide scans were performed 6 weeks postoperatively. Three weeks after the operation, the femoral heads showed evidence of osteonecrosis including increasing numbers of empty lacunae, decreased hematopoietic cells, and destroyed adipose tissue in the medullary cavity, which increased in severity at the subsequent follow-up evaluations at 6 and 9 weeks. Fractured trabeculae and fibrous tissue were noted 9 weeks postoperatively. Image analysis also revealed evidence of osteonecrosis, such as several osteopenic areas with sclerotic rims on the X-ray, several areas of low bone mineral density with sclerosis on the CT scan, increased uptake of the nuclide species in MRI, and an inhomogeneous long T2 signal on the radioisotopic images. Ethanol injection navigated by our novel template was successful in establishing a canine model of ONFH. This model can be used to test new treatment modalities for human ONFH.

The treatment effect of porous titanium alloy rod on the early stage talar osteonecrosis of sheep.

Osteonecrosis of the talus (ONT) may severely affect the function of the ankle joint. Most orthopedists believe that ONT should be treated at an early stage, but a concise and effective surgical treatment is lacking. In this study, porous titanium alloy rods were prepared and implanted into the tali of sheep with early-stage ONT (IM group). The curative effect of the rods was compared to treatment by core decompression (DC group). No significant differences in bone reconstruction were observed between the two groups at 1 month after intervention. After 3 months, the macroscopic view of gross specimens of the IM group showed ordinary contours, but the specimens of the DC group showed obvious partial bone defects and cartilage degeneration. Quantitative analysis of the reconstructed trabeculae by micro-CT and histological study suggested that the curative effect of the IM group was superior to that of the DC group at 3 months after intervention. These favorable short-term results of the implantation of porous titanium alloy rods into the tali of sheep with early-stage ONT may provide insight into an innovative surgical treatment for ONT.

[Preliminary study of gene expression profile associated with risk classification of childhood patients with acute lymphoblastic leukemia].

OBJECTIVE: To explore genes associated with risk classification of childhood acute lymphoblastic leukemia (ALL) by gene chip technology. METHODS: Group A and B were both composed of three newly diagnosed ALL cases with standard risk. After re-evaluation, group B was relegated to high-risk. The control group was composed of three idiopathic thrombocytopenic purpura (ITP) patients. The gene expression profiles of group A and B were studied by Illumina Human-6 Beadchip. Eighty-two ALL patients were selected as the experimental group and 21 with normal bone marrow as control group for real-time quantitative RT-PCR (RQ-PCR). RESULTS: (1) There were 19 genes expressed differently between group B and A, including 14 up-regulated as ABCC4 and BCL11A, 5 down-regulated genes as TOP2A. (2) ABCC4 and BCL11A were validated by RQ-PCR and their expression level was higher in the high risk group than in the standard risk group (P < 0.05). The gene expression level in the group A and B was higher than that in the normal control group (P < 0.01). TOP2A was also validated by RQ-PCR and its expression level in the high risk group was lower than that in the standard risk group (P < 0.05). The gene expression level in the groups A and B was lower than that in the normal control group and the difference was statistically significance (P < 0.01). (3) There was a significant difference in the expression level of ABCC4 between the remission and unremission patients (P < 0.05). There was no significant difference in the expression level of BCL11A between different clinical indicators (P > 0.05). There was significant difference in the expression level of TOP2A between remission and prednisone good responder groups (P < 0.05). CONCLUSIONS: Fourteen genes studied were involved in the pathogenesis and drug resistance mechanism in childhood ALL patients. Investigation of gene expression profile will be helpful for predicting drug resistance, prognosis, early intervention and target therapy in childhood ALL.

[Study on tumor necrosis factor-alpha and interleukin 2 in rat skin allograft].

OBJECTIVE: To elucidate the histocompatibility of tissue engineered rat skin through studying the effect of TNF-alpha and IL-2 in immunological rejection after rat skin allograft. METHODS: Tissue engineered skin that the basic materials were taken from neonatal SD rats was cultured in lab, grafted to adult Wistar rats. The expressions of TNF-alpha and IL-2 in grafted tissue were detected with immunohistochemical staining and Western blot. RESULTS: The expressions of TNF-alpha and IL-2 were remarkable in skin allograft group, but low in tissue engineered skin group. CONCLUSION: The expressions of TNF-alpha and IL-2 and the immunological rejection were closely related after skin allograft, but the tissue engineered skin has favorable histocompatibility and doesn't arose obvious immunological rejections.