The Dynamic Changes of AFP From Baseline to Recurrence as an Excellent Prognostic Factor of Hepatocellular Carcinoma After Locoregional Therapy: A 5-Year Prospective Cohort Study

BackgroundAlthough many studies have confirmed the prognostic value of preoperative alpha-fetoprotein (AFP) in patients with hepatocellular carcinoma (HCC), the association between AFP at baseline (b-AFP), subsequent AFP at relapse (r-AFP), and AFP alteration and overall survival in HCC patients receiving locoregional therapy has rarely been systematically elucidated.Patients and MethodsA total of 583 subjects with newly diagnosis of virus-related HCC who were admitted to Beijing You ‘an Hospital, Capital Medical University from January 1, 2012 to December 31, 2016 were prospectively enrolled. The influence of b-AFP, subsequent r-AFP, and AFP alteration on relapse and post-recurrence survival were analyzed.ResultsBy the end of follow-up, a total of 431 (73.9%) patients relapsed and 200 (34.3%) died. Patients with positive b-AFP had a 24% increased risk of recurrence compared with those who were negative. Patients with positive r-AFP had a 68% increased risk of death after relapse compared with those who were negative. The cumulative recurrence-death survival (RDS) rates for 1, 3, 5 years in patients with negative r-AFP were 85.6% (184/215), 70.2%(151/215), and 67.4%(145/215), while the corresponding rates were 75.1% (154/205), 51.2% (105/205), and 48.8% (100/205) in those with positive AFP (P<0.001). 35 (21.6%) of the 162 patients with negative b-AFP turned positive at the time of recurrence, and of this subset, only 12 (34.3%) survived. Of the 255 patients with positive b-AFP, 86 (33.7%) turned negative at the time of relapse, and of this subset, only 30 (34.9%) died. The 1-, 3-, and 5-year cumulative RDS rates were also compared among groups stratified by AFP at baseline and relapse. The present study found that patients with positive AFP at baseline and relapse, as well as those who were negative turned positive, had the shortest RDS and OS.ConclusionsNot only AFP at baseline but also subsequent AFP at relapse can be used to predict a post-recurrence survival, which can help evaluate mortality risk stratification of patients after relapse.

Machine Leaning Algorithm on Chemotherapeutic Drug Resistance Related Gene Classifier in Acute Myeloid Leukemia

Background: Acute myeloid leukemia (AML) is a heterogeneous disease in which 20-50% patients are resistant to chemotherapy. Relapsed/refractory (R/R) AML has a poor long-term prognosis resulting from chemotherapeutic drug resistance. However, the specific gene-drug pairs have been rarely reported. The purpose of our study was to explore an accurate algorithm for screening gene profiles classifier associated with chemotherapy drug sensitivity. Method: 43 AML patients from the Institute of Hematology and Blood Diseases Hospital were enrolled, including newly diagnosis (n=23) and R/R (n=20) AML between March 2019 to January 2020. We separated bone marrow cells (BMCs) of the 43 patients. Additionally, 24 therapeutic drugs panel (Figure 1) were selected and 7-concentration gradient were used to test drug sensitivity in vitro by HDS (High-throuphput Drug Sensitivity Analysis Strategy). Paired molecular data of RNA-seq and DNA methylation 850K CHIP implemented simultaneously. K-Means, an unsupervised algorithm, was applied to cluster all samples into two groups (Resistance/Sensitive group) according to IC50 and 100% PPC inhibition rate of each drug. We used a combination of different algorithms to build a proper gene classifier. The whole process was shown as Figure 2. The major objectives of the evaluating algorithm included F1 score, precision rate, recall rate and AUC. "pRRophetic", a R package, was modified to specify target type as "AML" in aim to estimate IC50 of samples based on GDSC database. Furthermore, we cultured Ara-C resistant cell line of HL-60 in purpose of comparing relative gene expression with wide type (wt) by quantitative Real-time PCR (qPCR) to prove the selected gene signature. Result: The HDS data suggested that sensitivities of newly diagnosis AML to chemotherapeutic-drugs were highly variable. However, Newly Diagnosis Group was more sensitive to majority chemotherapeutic drugs in contrast to R/R AML (P<0.05). As heterogeneous drug response in vitro, we found K-means (k=2) was more reasonable algorithm, as compared to K-means (k=3) for grouping all samples. SVM of transcription data for each drug showed a significantly advantage over other algorithms (P<0.001) with the median F1 score was 0.805295 (0.710648-0.934641), median AUC was 0.818 (0.545-1), median precision rate was 0.75 (0.55-1), and median recall rate was 0.76(0.66-1). The screening feature genes from SVM also performed well on different models (SVM, RF, logistics regression, KNN, Decision Tree) of test set. The SVM mean F1 scores for each algotithm were as follows:0.7382, 0.6128, 0.6530, 0.6168,and 0.5956. It is worth mentioning that SVM equally applies to DMP analysis (P<0.001).We confirmed our algorithm by using Ara-C selected genes grounded by SVM and RF. The difference of estimated IC 50 between resistant group and sensitive group based on SVM (P=0.058) was better than RF (p=0.087) (Figure 3). Compared to newly diagnosis AML, R/R Group possessed 2134 upregulated genes and 1210 down-regulated genes in the aspect of RNA-seq data. Crosslinking analysis of RNA-seq and methylation data of the two group, 1 gene were described as up-regulated expression with hypo-methylation, and 39 genes was down-regulated expression with hyper-methylation. 8/39 genes are corresponded with SVM algorithm. Moreover, 4-gene-drug pairs, including FOXC1 for anthracyclines drug, IGFBP5 for Ara-C, VTRNA1-1 and TKTL1 for pan-drug, were investigated by overlapping TOP 5 genes for each drug of SVM algorithm and DEGs between R/R and newly diagnosis AML. High expression of four genes was identified as a risk factor for AML prognosis. The result of qPCR revealed that IGFBP5 is overexpression in Ara-C-resistant HL-60 than wt. Conclusion: We construct a model of K-Means and RFE-DEG/DMP-SVM, a validated and precise computational approach, for predicting drug sensitivity related genes in AML patients. Up-regulated expression with hypo-methylation genes may be signature genes for drug resistance. 4-gene-based classifier may make contributions to chemotherapeutic-drug resistance prediction and AML treatment decision-making. Disclosures No relevant conflicts of interest to declare.

Serum Vitamin D Levels in Children with Immune Thrombocytopenia

Objective: Immune thrombocytopenia (ITP) is an acquired immuno-mediated disorder characterized by thrombocytopenia with an increased risk of bleeding. In recent years 1,25[OH]2D3 has been rediscovered as an immune modulator. We decided to evaluate serum Vitamin D levels in a cohort of children with immune thrombocytopenia in order to discover if Vitamin D concentrations may predict ITP duration. Methods: Thirty children were enrolled in this study (sixteen with chronic ITP and fourteen with newly diagnosed ITP) to assess serum Vitamin D levels. Results:: The results showed that 80% of the enrolled children presented a D hypovitaminosis status. Children with newly diagnosis ITP showed no statistically significantly higher median values of Vitamin D compared to chronic ITP. Conclusions:: This study may suggest that Vitamin D deficiency does not represent a chronicity factor for ITP. However, further studies are needed to understand the role of Vitamin D in ITP pathogenesis.

Comparison Study of the Effect of Erlotinib as a Tyrosine Kinase Inhibitor on Electrolyte Levels in Type2 Diabetic and Diabetic Nephropathy

Diabetes mellitus can be defined as a metabolic disorder disease .Complication of diabetes are due to diabetic nephropathy .This study was done in vitro to study the effect of different concentrations of erlotinib inhibitor ( tyrosine kinase inhibitor) on electrolyte levels (Mg⁺²,Ca⁺²,Na⁺) in sera of Iraqi patients with newly diagnosis type2 diabetes and diabetic nephropathy in addition to find the best percentage inhibition for utilizing different concentrations from erlotinib (6.97x10⁻⁷, 9.30x10⁻⁷ ,1.16x10⁻⁶,1.39x10⁻⁶ ,1.62x10⁻⁵ )molar on electrolyte levels . This study was conducted in The National Diabetes Center, Al-Mustansiriya University - Baghdad and included 150 patients divided into50 patients type 2 diabetic as group (G2) , 50patients diabetic nephropathy as group(G3) and also 50 healthy as control group(G1).The period time for aggregation the blood sampling was from July to October 2017 . All patients were within (18 to 60) years age. Erlotinib(Tyrosine kinase inhibitor) affected on serum Mg ⁺²levels in human as a mild effect and a slight effect on seum Na⁺ and Ca⁺².The best inhibition of erlotinib in concentration (1.62x10⁻5)M for both serum Na⁺ and Ca⁺² in newly diagnosis diabetes type 2 and diabetic nephropathy.serum Mg ⁺²levels showed best inhibition in concentration (9.30x10⁻7)M