Prognostic stratification of sepsis through DNA damage response based RiskScore system: insights from single-cell RNA-sequencing and transcriptomic profiling.

Background: A novel risk scoring system, predicated on DNA damage response (DDR), was developed to enhance prognostic predictions and potentially inform the creation of more effective therapeutic protocols for sepsis. Methods: To thoroughly delineate the expression profiles of DDR markers within the context of sepsis, an analytical approach utilizing single-cell RNA-sequencing (scRNA-seq) was implemented. Our study utilized single-cell analysis techniques alongside weighted gene co-expression network analysis (WGCNA) to pinpoint the genes that exhibit the most substantial associations with DNA damage response (DDR). Through Cox proportional hazards LASSO regression, we distinguished DDR-associated genes and established a risk model, enabling the stratification of patients into high- and low-risk groups. Subsequently, we carried out an analysis to determine our model's predictive accuracy regarding patient survival. Moreover, we examined the distinct biological characteristics, various signal transduction routes, and immune system responses in sepsis patients, considering different risk categories and outcomes related to survival. Lastly, we conducted experimental validation of the identified genes through in vivo and in vitro assays, employing RT-PCR, ELISA, and flow cytometry. Results: Both single-cell RNA sequencing (scRNA-seq) and bulk transcriptomic analyses have demonstrated a strong correlation between DNA damage response (DDR) levels and sepsis prognosis. Specific cell subtypes, including monocytes, megakaryocytes, CD4+ T cells, and neutrophils, have shown elevated DDR activity. Cells with increased DDR scores exhibited more robust and numerous interactions with other cell populations. The weighted gene co-expression network analysis (WGCNA) and single-cell analyses revealed 71 DDR-associated genes. We developed a four-gene risk scoring system using ARL4C, CD247, RPL7, and RPL31, identified through univariate COX, LASSO COX regression, and log-rank (Mantel-Cox) tests. Nomograms, calibration plots, and decision curve analyses (DCA) regarding these specific genes have provided significant clinical benefits for individuals diagnosed with sepsis. The study suggested that individuals categorized as lower-risk demonstrated enhanced infiltration of immune cells, upregulated expression of immune regulators, and a more prolific presence of immune-associated functionalities and pathways. RT-qPCR analyses on a sepsis rat model revealed differential gene expression predominantly in the four targeted genes. Furthermore, ARL4C knockdown in sepsis model in vivo and vitro caused increased inflammatory response and a worse prognosis. Conclusion: The delineated DDR expression landscape offers insights into sepsis pathogenesis, whilst our riskScore model, based on a robust four-gene signature, could underpin personalized sepsis treatment strategies.

Effect of interfering IGF-1R by siRNA on cell cycle and apoptosis of hypoxic hepatocellular carcinoma HepG2 cells / 中国肿瘤生物治疗杂志

@# Objective: To explore the effect of interfering insulin-like growth factors-1 receptors (IGF-1R) by small interfering RNA (siRNA) on cell cycle and apoptosis of hypoxic hepatocellular carcinoma HepG2 cells. Methods: The hypoxic hepatocellular carcinoma model was established via cobalt chloride treatment. Three siRNAs targeting IGF1R gene and one negative control siRNA were designed and synthesized. They were transfected into hypoxic HepG2 cells, and 24 h later, the transfection efficiency was detected by fluorescent microscopy. The protein expression of IFG-1R was detected with Western blotting (WB) to screen the siRNA with highest transfection efficacy. The selected siRNA was used to transfect hypoxic HepG2 cells. The proliferation of hypoxic HepG2 cells was determined by MTT assay. Cell cycle distribution and apoptosis were analyzed by Flow cytometry. WB was performed to detect the proteinexpressionsofCDK1,CDK2andCaspase-3inHepG2cells. Results: The hypoxic hepatocellular carcinoma model was successfully established. IGF-1R-siRNA-2 showed the most effective interference efficiency and the most significant knockdown of IGF-1R (all P<0.01). The proliferation of HepG2 cells transfected with IGF-1R siRNA-2 was significantly suppressed (P<0.05 or P<0.01), the cell cycle was blocked at G0/G1 phase (P<0.05), and the apoptosis rate was increased up to (25.3±1.3)% P<0.01). In the meanwhile, the expressions of CDK1 and CDK2 were decreased and the expression of Caspase-3 was increased in hypoxic HepG2 cells after IGF-1R knockdown (P<0.05). Conclusion: Interfering IGF-1R by siRNA inhibits the malignant biological behaviors of hypoxic HepG2 cells via regulating cell cycle and apoptosis-related proteins. IGF-1R may be a potential target for the treatment of HCC.

Effect of curcumin on angiogenesis of rats with DEN-induced hepatocellular carcinoma under hypoxia / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of curcumin on angiogenesis in rats with DEN (diethylnitrosamine)-induced HCC (hepatocellular carcinoma) under hypoxia. Methods: Rat HCC was induced by DEN, and its hepatic hypoxia model was established by ligating hepatic artery. The rats with established HCC model were randomly divided into four groups according to digital table method: lipiodol embolization group (group A), lipiodol combined with curcumin embolization group (group B), lipiodol combined with peripheral liver capsule group (group C), lipiodol combined with curcumin and peripheral liver capsule group (group D), with 10 rats in each group. VEGF expression in HCC cells and tissues, microvessel density (MVD), and median survival time (MST) of rats in each group were compared. Results: VEGF protein expression and microvessel density in group B, D were significantly lower than those in A group ( P <0.01), while those in C group had no significant difference compared with group A ( P >0.05). MST in group B, C and D was significantly longer than that in group A ( P <0.05), and the MST in group D was higher than that in group B and C ( P <0.05). Conclusion: Curcumin can inhibit tumor angiogenesis and decrease VEGF expression and MVD in HCC rats under hypoxia, thus further prolong the survival time of the rats.

A large primary retroperitoneal vaginal leiomyosarcoma: a case report.

INTRODUCTION: Primary vaginal leiomyosarcomas are uncommon, especially those growing outside the vagina.Out of all malignant vaginal neoplasms, leiomyosarcomas account for about 2%. Reports in the literature mostly concern the pathology of these tumors; few reports have been published that discuss how to surgically remove them. CASE PRESENTATION: A 69-year-old Chinese woman presented with a mass in her buttocks that had been present for more than four months. Computed tomography demonstrated a mass of approximately 12.0×9.5×8.0cm in her retroperitoneal space. We resected the tumor via a posterior incision, and resected part of her sacrum and coccyx. The resected tumor was diagnosed by its pathological features as a leiomyosarcoma. Our patient received adjuvant chemotherapy after surgery. She was free of disease at a one-year follow-up and her general condition is good. CONCLUSIONS: We report a rare case of a primary vaginal leiomyosarcoma that was resected through an approach that has not, to the best of our knowledge, been previously reported. This case report adds valuable knowledge to the sparse available literature on the surgical treatment of vaginal leiomyosarcomas.

Effect of hypoxia on hypoxia inducible factor-1α, insulin-like growth factor I and vascular endothelial growth factor expression in hepatocellular carcinoma HepG2 cells.

Hypoxic microenvironments and angiogenesis have been a focus of tumor research in previous years. The aim of the the present study was to create a hypoxic model and observe the effect of hypoxia on the expression of hypoxia inducible factor-1α (HIF-1α), insulin-like growth factor I (IGF-1) and vascular endothelial growth factor expression. The hypoxia model was generated using cobalt chloride (CoCl2) and an MTT assay was used to observe the influence of hypoxia on HepG2 cells. Reverse transcription-polymerase chain reaction, western blotting, ELISA and confocal immunofluorescence microscopy were used to detect the expression of HIF-1α, IGF-1 and VEGF in HepG2 cells, in which hypoxia was induced by various concentrations of CoCl2 and for various incubation times. The cell viability worsened with increasing concentrations of CoCl2. The expression of HIF-1α and IGF-1R was observed in hypoxic HepG2 cells, with the exception of HIF-1α mRNA. The expression of IGF-1R and VEGF mRNA and protein was correlated with the concentration of CoCl2 and the time that hypoxia was induced for. The expression of HIF-1α mRNA and protein was positively correlated with the expression of the VEGF mRNA and protein in a dose- and time-dependent manner under hypoxic conditions. Using immunofluorescence, it was observed that IGF-1R and HIF-1α were secreted from the hypoxic HepG2 cells. It was concluded that hypoxia induces the accumulation of IGF-1R and HIF-1α mRNA and protein, which regulates the expression of VEGF mRNA and protein in hypoxic HepG2 cells.

Variant arterial supply to the lesser curvature of the stomach and duodenum from double inferior phrenic arteries.

A rare case of the left accessory inferior phrenic artery (IPA) supplying the lesser curvature of the stomach and the right accessory IPA supplying the duodenum was observed during interventional radiography. To our knowledge, variation arteries supplying the stomach and duodenum have never been reported in English literature; herein, we report the first case of variant arterial supply to the lesser curvature of the stomach and duodenum from double IPAs.

Perineal lipoblastoma: a case report and review of literature.

Lipoblastoma is a rare benign mesenchymal tumor that composes of embryonal white fat tissue and typically occurs in infants or young children under 3 years of age. It usually affects the extremities, trunk, head, and neck. The perineum is a rare location with only 7 cases reported in the literature. We describe a case of 3-year-old girl with a lipoblastoma arising from perineum. An approximately 4.5 cm × 3.5 cm × 2.5 cm nodule was resected in left perineum with satisfied results. Pathological examination showed that it was composed of small lobules of mature and immature fat cells, separated by fibrous septa containing small dilated blood vessels. The left perineal lipoblastoma, although rare, should be differentiated from some other mesenchymal tumors with similar histologic and cytological features.