ABSTRACT
Background: Point of care ultrasound (POCUS) is a cost-effective, non-invasive procedure with high diagnostic reliability and therapeutic utilities. For various reasons, it is not being used routinely in neurosurgical intensive care unit (ICU). We have introduced a systematic use of POCUS in critically ill patients in our neurosurgical ICU. We have studied the various indications and benefit of using this technique. Methods: This is a prospective, single center cohort observational study done in patients who were admitted in a tertiary neurosurgical ICU over 1 year (17th September 2020 to 16th September 2021). POCUS was used daily as per a standardized format for multiple purposes. A formal training to the operator was provided and standardized method of evaluation and intervention was used. Outcome was studied to understand the impact of the POCUS and difficulties encountered during its use. Results: POCUS was used in 240 patients, including all patients for primary and secondary surveys, 192 patients (80%) for optic nerve sheath diameter (ONSD) measurement, 14 myelomeningoceles for trans-fontanelle ultrasound study, 16 post operative cases of aneurysm clipping for transcranial Doppler (TCD) study, 86 patients for deep vein thrombosis (DVT) screening, 17 for evaluation of ventriculo-peritoneal (VP) shunt functionality, 30 for transcranial defect ultrasound, 45 for chest ultrasound, 4 for evaluation of hemodynamic shock and 67 patients who had difficult cannualtion or while insertion of central venous catheter placement. POCUS was also used for difficult cannulation, central catheter placement and e-FAST scan. Significant findings were reported 129 times, which led to immediate change in management in 62 patients (25.83%) as compared to 16 patients in whom significant findings were not reported using POCUS, but management was changed using other radiological modalities (P<0.01). There was initial lag in adopting the technique, however with practice, the team developed confidence. As a result, the accuracy and time taken to perform the procedure was reduced significantly. Conclusions: Routine systematic use of POCUS can be beneficial not only for the rapid diagnosis and prompt management of patients, but also helpful in monitoring and performing various procedures in neurosurgical ICU. Though not all modalities mentioned in standardized format was used in all patients, use of this format has helped improved training and maintain proper use of POCUS in our ICU.
ABSTRACT
BACKGROUND/AIM: Cancer-associated fibroblasts (CAFs) are important factors in the progression of hepatocellular carcinoma (HCC). But the characterization of these cells remains incomplete. This study aims to identify a panel of markers for CAFs that are associated with HCC progression. MATERIALS AND METHODS: The sequencing data and clinicopathological characteristics of 366 patients were obtained from the Cancer Genome Atlas (TCGA) database (366 HCC tissues and there were 50/366 cases with corresponding normal liver tissues). In vitro validation of the markers was performed by quantitative real-time PCR using the hepatic stellate cell line LX2 induced by the HCC cell line Huh7. The activation of LX2 was confirmed by α-smooth muscle actin and fibroblast activation protein, using quantitative real-time PCR and immunofluorescence staining. In vivo detections of the 12 markers were done in 40 tissue samples (30 HCC and 10 normal). RESULTS: We successfully identified 12 CAF markers from TCGA data: FGF5, CXCL5, IGFL2, MMP1, ADAM32, ADAM18, IGFL1, FGF8, FGF17, FGF19, FGF4, and FGF23. The 12-marker panel was associated with the pathological and clinical progressions of HCC. All 12 markers were upregulated in vitro. In vivo expressions of these markers were paralleled with those in TCGA data. CONCLUSION: A 12-marker panel of CAFs in HCC is identified, which is associated with both pathological and clinical progressions of cancer.
ABSTRACT
Fibroblast activation protein (FAP) is a serine protease that has been reported in fibroblasts and some carcinoma cells, which correlates with poor patient outcomes. FAP can be induced under hypoxia which is also vital in the malignant behaviors of cancer cells. However, the role of FAP and its correlation with hypoxia has not been investigated in HCC cancer cells. In tissues from post-surgical HCC patients in our center, we adopted immunohistochemistry staining (IHC), western blot and quantitative RT-PCR to detect the expression levels of FAP and the hypoxia related marker, hypoxia inducible factor 1α (HIF-1α). X-tile software was used for the determination of high and low expression of FAP and HIF-1α after the IHC analysis. Clinicopathological analysis, Kaplan-Meier analysis and Cox regression model were performed. In-vitro experiments were performed to confirm the relationship between FAP and hypoxia in HCC cancer cell lines (HepG2, Huh7 and MHCC97H). Results revealed that expression levels of FAP and HIF-1α were significantly correlated (Pearson r2 = 0.2753, p < 0.0001) in IHC analysis of the 138-patient cohort. Western blot and quantity RT-PCR indicated parallel changes in 11 post-surgical fresh frozen tissues. The HIF-1α and FAP expression were associated with serum AFP, TNM, tumor size and vascular invasion. Cox regression analysis showed that HIF-1α/ FAP combination were the independent predictor for overall survival (OS) and time-to-recurrence (TTR) in post-surgical HCC patients. Kaplan-Meier analyses revealed that the patient with high levels of HIF-1α, FAP and combined HIF-1α/FAP had the shortest OS and TTR. In-vitro experiments showed that FAP was increased in hypoxic HCC cancer cell lines in parallel with that of HIF-1α and three EMT markers (E-cadherin, Snail and TWIST). In conclusion, the up-regulation of FAP in HCC cancer cells under hypoxia can be indicative of poor prognosis in patients.