Clinical dose rationale of tislelizumab in patients with solid or hematological advanced tumors.

Tislelizumab, an anti-programmed cell death protein 1 monoclonal antibody, has demonstrated improved survival benefits over standard of care for multiple cancer indications. We present the clinical rationale and data supporting tislelizumab dose recommendation in patients with advanced tumors. The phase I, first-in-human, dose-finding BGB-A317-001 study (data cutoff [DCO]: August 2017) examined the following tislelizumab dosing regimens: 0.5-10 mg/kg every 2 weeks (q2w), 2-5 mg/kg q2w or q3w, and 200 mg q3w. Similar objective response rates (ORRs) were reported in the 2 and 5 mg/kg q2w or q3w cohorts. Safety outcomes (grade ≥3 adverse events [AEs], AEs leading to dose modification/discontinuation, immune-mediated AEs, and infusion-related reactions) were generally comparable across the dosing range examined. These results, alongside the convenience of a fixed q3w dose, formed the basis of choosing 200 mg q3w as the recommended dosing regimen for further clinical use. Pooled exposure-response (E-R) analyses by logistic regression using data from study BGB-A317-001 (DCO: August 2020) and three additional phase I/II studies (DCOs: 2018-2020) showed no statistically significant correlation between tislelizumab pharmacokinetic exposure and ORR across multiple solid tumor types or classical Hodgkin's lymphoma, nor was exposure associated with any of the safety end points evaluated over the dose range tested. Hence, tislelizumab showed a relatively flat E-R relationship. Overall, the totality of data, including efficacy, safety, and E-R analyses, together with the relative convenience of a fixed q3w dose, provided clinical rationale for the recommended dosing regimen of tislelizumab 200 mg q3w for multiple cancer indications.

[Ecological network optimization of Jiuquan City, Gansu, China based on complex network theory and circuit model]. / åŸºäºŽå¤æ‚ç½‘ç»œç†è®ºå’Œç”µè·¯æ¨¡åž‹çš„é ’æ³‰å¸‚ç”Ÿæ€ç½‘ç»œä¼˜åŒ–.

Building a scientific and reasonable ecological network is the key for optimizing the pattern of territorial development and protection, and is of great significance for ensuring regional ecological security and promoting the virtuous cycle of ecosystems. In previous studies, nodal attack method (destruction of ecological source area) was often used in the "robustness" evaluation of ecological networks. Actually, the ecological corridor is more fragile than the source area, and thus the nodal attack method is not reasonable. In this study, taking Jiuquan City as the research area, based on the circuit model to construct the ecological network, we carried out the topology optimization of ecological network by using three strategies (random edge increase, node degree and priority edge increase with low node intermedium number) in complex network theory. We compared and analyzed the "robustness" of ecological network before and after optimization by constructing edge attack strategy, and selected the best network optimization strategy. The results showed that 65 ecological source areas were identified in Jiuquan City, with a total area of 20275.15 km2, and that grassland accounted for 89.5% of the source area. We identified 179 ecological corridors with a total length of 6387.16 km, 158 ecological barrier points with a total area of 1385.5 km2. The unused land accounted for 92.2% of the total barrier points area. We identified 63 ecological pinch points, mainly concentrated in the source edge and corridor intersection. Among them, the spatial distribution of 11 barrier points and pinch points was consistent, which was the key area to be repaired in ecological network optimization. The three optimization strategies had significantly improved the stability of ecological network in Jiuquan City. The relative size of the maximum connected subgraph and the edge connected rate of the ecological network of the optimization strategy of adding edges according to degree were all the most stable under random attack mode and deliberate attack mode, which was the best optimization scheme for ecological network in Jiuquan City.

[Diagnosis of a case with Hermansky-Pudlak syndrome type 5 through high-throughput sequencing and a literature review].

OBJECTIVE: To explore the clinical and genetic characteristics of a patient with Hermansky-Pudlak syndrome type 5 (HPS-5). METHODS: A child with HPS-5 who had attended the Children's Hospital Affiliated to Shandong University on October 3, 2019 was selected as the study subject. Clinical data of the child were collected. Genetic variant was analyzed through high-throughput sequencing. A literature review was also carried out. RESULTS: The child, a 1-year-and-5-month-old girl, had nystagmus since childhood, lost of retinal pigmentation by fundus examination and easy bruising. High-throughput sequencing revealed that she has harbored compound heterozygous variants of the HPS5 gene, namely c.1562_1563delAA (p.F521Sfs*27) and c.1404C>A (p.C468X), which were inherited from his father and mother, respectively. Based on the guidelines from the American College for Medical Genetics and Genomics (ACMG), both variants were predicted to be pathogenic (PVS+PM2_Supporting+PM3+PP4). Among 18 previously reported HPS-5 patients, all had had eye problems, and most of them had tendency for bleeding. Eight cases had carried compound heterozygous variants of the HPS5 gene, 8 carried homozygous variants, 2 carried double homozygous variants, and most of them were null mutations. CONCLUSION: The c.1562_1563delAA(p.F521Sfs*27) and c.1404C>A (p.C468X) compound heterozygous variants of the HPS5 gene probably underlay the HPS-5 in this child. High-throughput sequencing has provided an important tool for the diagnosis. HSP-5 patients usually have typical ocular albinism and/or oculocutaneous albinism and tendency of bleeding, which are commonly caused by compound heterozygous and homozygous variants of the HPS5 gene, though serious complications have been rare.

Mining and modification of Oryza sativa-derived squalene epoxidase for improved ß-amyrin production in Saccharomyces cerevisiae.

ß-Amyrin is a pentacyclic triterpenoid and has anti-viral, anti-bacterial and anti-inflammatory activities. The synthetic pathway of ß-amyrin has been analyzed and its heterogeneous synthesis has been achieved in Saccharomyces cerevisiae. Squalene epoxidase (SQE) catalyzes the oxygenation of squalene to form 2,3-oxidosqualene and is rate-limiting in the synthetic pathways of ß-amyrin. The endogenous SQE in S. cerevisiae is insufficient for high production of ß-amyrin. Herein, eight squalene epoxidases derived from different plants were selected and characterized in S. cerevisiae for improved biosynthesis of ß-amyrin. Among them, the squalene epoxidase from Oryza sativa (OsSQE52) showed the best performance compared to other plant-derived sources. Through protein remodeling, the mutant OsSQE52L256R, obtained based on modeling analysis, increased the titer of ß-amyrin by 2.43-fold compared to that in the control strain with ERG1 overexpressed under the same conditions. Moreover, the expression of OsSQE52L256R was optimized with the improvement of precursor supply to further increase the production of ß-amyrin. Finally, the constructed strains produced 66.97 mg/L ß-amyrin in the shake flask, which was 6.45-fold higher than the original strain. Our study provides alternative SQEs for efficient production of ß-amyrin as well as other triterpenoids derived from 2,3-oxidosqualene.

[Enrichment Characteristics of Heavy Metals and Health Risk in Different Vegetables].

The health risk caused by heavy metal accumulation in vegetables is of great concern. In this study, a database of heavy metal content in a vegetable-soil system in China was constructed through literature review and field sample collection. A systematic analysis of seven heavy metal contents in edible parts of vegetables and their bioaccumulation capacity among different vegetables was also performed. Additionally, the non-carcinogenic health risks of four types vegetables were assessed by using Monte Carlo simulation (MCS). The mean values of Cd, As, Pb, Cr, Hg, Cu, and Zn in the edible parts of the vegetables were 0.093, 0.024, 0.137, 0.118, 0.007, 0.622, and 3.272 mg·kg-1, and the exceedance rates of the five toxic elements were:Pb (18.5%)>Cd (12.9%)>Hg (11.5%)>Cr (4.03%)>As (0.21%). Leafy vegetables showed high Cd enrichment, and root vegetables showed high Pb enrichment, with mean bioconcentration factors of 0.264 and 0.262, respectively. Generally, legumes vegetables and solanaceous vegetables showed lower bioaccumulation for heavy metals. The health risk results indicated that the non-carcinogenic risk for single elements of vegetable intake was within the acceptable range, with the health risk for children being higher than that for adults. The mean non-carcinogenic risk for single elements were:Pb>Hg>Cd>As>Cr. The multi-element combined non-carcinogenic risks of four types vegetables were:leafy vegetables>root vegetables>legume vegetables>solanaceous vegetables. Planting lower-heavy metal bioaccumulation vegetables in heavy metal-contaminated farmland is an effective method to minimize the health risk.

[Clinical characteristics and genetic analysis of two children with Autosomal dominant mental retardation type 21 due to variants of CTCF gene].

OBJECTIVE: To explore the clinical and genetic characteristics of two children with developmental delay. METHODS: Two children who had presented at the Children's Hospital Affiliated to Shandong University on August 18, 2021 were enrolled as the study subjects. Clinical and laboratory examination, chromosomal karyotyping and high-throughput sequencing were carried out for both children. RESULTS: Both children had a 46,XX karyotype. High-throughput sequencing showed that they have respectively carried a c.489delG (p.Q165Rfs*14) and a c.1157_1158delAT (p.Y386Cfs*22) frameshifting variant of the CTCF gene, both had a de novo origin and were unreported previously. CONCLUSION: The CTCF gene variants probably underlay the development delay in the two children. Above discovery has enriched the mutational spectrum of the CTCF gene and has important implications for revealing the genotype-phenotype correlation for similar patients.

Model-based population pharmacokinetic analysis of tislelizumab in patients with advanced tumors.

Tislelizumab, a humanized immunoglobulin G4 monoclonal antibody, is a programmed cell death protein 1 (PD-1) inhibitor designed to minimize Fc gamma receptor binding on macrophages to limit antibody-dependent phagocytosis, a potential mechanism of resistance to anti-PD-1 therapy. The pharmacokinetic (PK) profile of tislelizumab was analyzed with population PK modeling using 14,473 observed serum concentration data points from 2596 cancer patients who received intravenous (i.v.) tislelizumab at 0.5-10 mg/kg every 2 weeks or every 3 weeks (q3w), or a 200 mg i.v. flat dose q3w in 12 clinical studies. Tislelizumab exhibited linear PK across the dose range tested. Baseline body weight, albumin, tumor size, tumor type, and presence of antidrug antibodies were identified as significant covariates on central clearance, whereas baseline body weight, sex, and age significantly affected central volume of distribution. Sensitivity analysis showed that these covariates did not have clinically relevant effects on tislelizumab PK. Other covariates evaluated, including race (Asian vs. White), lactate dehydrogenase, estimated glomerular filtration rate, renal function categories, hepatic function measures and categories, Eastern Cooperative Oncology Group performance status, therapy (monotherapy vs. combination therapy), and line of therapy did not show a statistically significant impact on tislelizumab PK. These results support the use of tislelizumab 200 mg i.v. q3w without dose adjustment in a variety of patient subpopulations.

Elevated pretreatment neutrophil-to-lymphocyte ratio indicate low survival rate in apatinib-treated patients with non-small cell lung cancer: A STROBE-compliant article.

This study aimed to analyze the predictive value of the neutrophil-to-lymphocyte ratio (NLR) to better clarify which patients with advanced non-small cell lung cancer (NSCLC) would benefit most from apatinib after multiline treatment for drug resistance. This observational cohort study involved patients with advanced NSCLC who were treated with apatinib between May 2016 to May 2018. The participants in this study had previously been treated with at least two treatment regimens. Multivariate logistic regression and Cox proportional risk models were used to evaluate the overall survival (OS) and progression-free survival (PFS) of the pretreatment NLR. A total of 125 patients were reviewed. The median age was 64 years (range, 33-92); and 32.8% of the patients were female. Only 0.8% of the patients had an Eastern Cooperative Oncology Group Performance Status (ECOG-PS) score ≥ 2. In multivariate analysis, pretreatment NLR ≥ 5 had an independent correlation with inferior OS (median 2.07 vs 3.40 months; HR 1.493, 95% CI 1.022-2.182; P = .038) and inferior PFS (median 1.83 vs 2.76 months; HR 1.478, 95% CI 1.015-2.153; P = .042). Elevated pretreatment NLR is associated with shorter OS and PFS in patients with advanced NSCLC treated with apatinib after multiline treatment for drug resistance.

Evolution of Preset Void and Damage Characteristics in Aluminum during Shock Compression and Release.

It is well known that initial defects play an essential role in the dynamic failure of materials. In practice, dynamic tension is often realized by release of compression waves. In this work, we consider void-included single-crystal aluminum and investigate the damage characteristics under different shock compression and release based on direct atomistic simulations. Elastic deformation, limited growth and closure of voids, and the typical spall and new nucleation of voids were all observed. In the case of elastic deformation, we observed the oscillatory change of void volume under multiple compression and tension. With the increase of impact velocity, the void volume reduced oscillations to the point of disappearance with apparent strain localization and local plastic deformation. The incomplete or complete collapsed void became the priority of damage growth under tension. An increase in sample length promoted the continuous growth of preset void and the occurrence of fracture. Of course, on the release of strong shock, homogeneous nucleation of voids covered the initial void, leading to a wider range of damaged zones. Finally, the effect of the preset void on the spall strength was presented for different shock pressures and strain rates.

The risks of sulfur addition on cadmium accumulation in paddy rice under different water-management conditions.

Recently, the application of sulfur (S) has been recommended to control the accumulation of cadmium (Cd) in rice in contaminated paddy soil. However, the effects of exogenous S on Cd transfer in paddy rice systems under different water-management practices have not been systematically investigated. Pot experiments were performed to monitor the composition of soil pore water and the Cd accumulation in iron plaque and rice tissue were compared under different S (0 and 200 mg/kg Na2SO4) and water (continuous and discontinuous flooding) treatments. Sulfur application significantly increased Cd concentrations in soil pore water under discontinuous flooding conditions, but slightly reduced them under continuous flooding. Moreover, the oxidation/reduction potential (Eh) was the most critical factor that affected the Cd levels. When the Eh exceeded -42.5 mV, S became the second critical factor, and excessive S application promoted Cd dissolution. In addition, S addition elevated the Cd levels in iron plaque and reduced the Cd transfer from the iron plaque to rice roots. In rice, S addition inhibited Cd transfer from the rice roots to the straw; thus, more Cd was stored in the rice roots. Nevertheless, additional S application increased the Cd content in the rice grains by 72% under discontinuous flooding, although this effect was mitigated by continued flooding. Under simulated practical water management conditions, S addition increased the risk of Cd contamination in rice, suggesting that S application should be reconsidered as a paddy fertilization strategy.

Mutational Characteristics of Causative Genes in Chinese Hereditary Spherocytosis Patients: a Report on Fourteen Cases and a Review of the Literature.

Background: Hereditary spherocytosis (HS), characterized by the presence of spherocytic red cells in peripheral blood, hemolysis, splenomegaly, jaundice, and gallstones, is a common form of inherited hemolytic anemia (HA). To date, five causative genes associated with HS have been identified, including ANK1, SPTB, SPTA1, SLC4A1, and EPB42. Methods: Clinically suspected patients with HS or undiagnosed HA from 14 Chinese families were enrolled in this study. We presented the patients' clinical features and identified the causative gene variants in these patients using whole exome sequencing (WES), with 10 novel and four reported mutations in the ANK1 and SPTB genes (seven mutations in ANK1 and seven in SPTB), individually. Then, we reviewed all available literature on Chinese HS patients from 2000 to 2020 in PubMed and Chinese Journals with genetic results and clinical information, to delineate gene mutation spectrum and potential correlation with phenotypes. Results: A total of 158 variants (including 144 in previous reports and 14 in this study) indicated that ANK1 (46%) and SPTB (42%) were the most frequently mutated genes in Chinese HS patients, followed by SLC4A1 (11%) and SPTA1 (1%), while no mutations in EPB42 was reported. Most of the mutations in ANK1 and SPTB were nonsense (26/73 in ANK1 and 32/66 in SPTB) and frameshift (20/73 in ANK1 and 15/66 in SPTB), while missense mutations (14/18) accounted for the majority in SLC4A1. The higher mutation frequency of ANK1 was found in its exon 8, 9, 26, and 28. The majority of mutations in SPTB were located in its exon 13, 15, and 18-30, whereas mutations in SLC4A1 were scattered throughout the entire region of the gene. Conclusion: Our study expanded the mutation spectrum of ANK1 and SPTB. Furthermore, we clarified the mutational characteristics of causative genes by reviewing all available literature on Chinese patients with HS.

Cadmium accumulation in wheat and maize grains from China: Interaction of soil properties, novel enrichment models and soil thresholds.

The cadmium (Cd) activity in soil has been widely studied. However, the interactive effects of soil properties (e.g. soil pH, CEC, and SOM) on Cd transfer from soil to grain are generally overlooked. In total 325 datasets including soil pH, CEC, SOM, and soil Cd content were used in this study. The descriptive statistics indicated that Cd content in wheat and maize soils ranged from 0.05 to 10.31 mg/kg and 0.02-13.68 mg/kg, with mean values of 0.87 and 1.14 mg/kg, respectively. Cd contents in wheat and maize grains were 0.01-1.36 mg/kg and 0.001-1.08 mg/kg with average values of 0.15 and 0.10 mg/kg, respectively. The results of SEM demonstrated that the interactive effects of soil properties contributed more to Cd transfer from soil to wheat grain than the soil Cd content. Subsequently, CITs-MLR indicated that the critical factors, including soil pH and total soil Cd content, could mask the contribution of other soil properties on Cd accumulation in grain; soil CEC may prevent Cd from leaching and therefore improve grain Cd level of wheat especially at acidic soil condition. The result of derived Cd thresholds revealed that current Cd thresholds are not completely suitable to wheat and maize grain at different soil conditions. This study provides a new model for further investigation on relationships between soil properties, soil Cd content and grain Cd level.

Arsenic transfer and accumulation in the soil-rice system with sulfur application and different water managements.

Rice (Oryza sativa L.) can readily accumulate arsenic (As), owing to its high capacity to take up As and special flooding cultivation, which poses a potential risk to human health. Although sulfur (S) can influence As accumulation in rice, its behavior in the rice-soil system is not clear under practical water management conditions. In this study, the transfer and soil solution dynamics of As in the whole soil-rice system was investigated under continuous flooding or intermittent drainage, either with S addition or not. The results showed that intermittent drainage effectively reduced As concentration in soil solution regardless of S conditions, and it only reduced As content in rice without S addition. Exogenous S decreased As concentration in soil solution and rice, except in the roots of mature rice, under continuous flooding. Sulfur addition significantly decreased the total As and As(III) contents in rice grains, by 62% and 79% under continuous flooding and by 50% and 76% under intermittent drainage, respectively. Moreover, the addition of S resulted in impaired iron plaque and inhibited binding of As. Sulfur also reduced As translocation from rice roots to shoots. Therefore, S could alleviate the crisis of excessive accumulation of As in rice grains caused by flooded environment through various adjustments to the soil-rice system.

[Analysis of clinical and genetic characteristics of a child with ring chromosome 4 syndrome].

OBJECTIVE: To explore the genetic basis for a child featuring short stature. METHODS: G-banded karyotyping, chromosomal microarray analysis (CMA) and high-throughput sequencing were carried out on peripheral blood sample from the child. RESULTS: The karyotype of the child was ascertained as 45,XY,-4[3]/46,XY,r(4)(p16q35)[84]/47,XY,-4,r(4)(p16q25)*2[7]/48,XY,-4,r(4)(p16q35)*3[1]/46,XY,dic r(4;4)(p16q35;p16q35)[2]/46,XY,add(4)(p16)[3]. A 647 kb deletion at 4p16.3 was identified by CMA, which encompassed 6 OMIM genes including ZNF141, PIGG, PDE6B, ATP5I, PCGF3 and MYL5. High-throughput sequencing has identified no pathogenic/likely pathogenic variants consistent with the clinical symptoms. CONCLUSION: A rare ring chromosome 4 syndrome was identified by combined chromosomal karyotyping, CMA and high-throughput sequencing. Conventional cytogenetic analysis and genetic testing in combine have enabled the diagnosis in this case.

Structural Evolution and Optical Property Tunability by Halogen Substitution in [N(CH3)4]MX2 (M = Ga+, In+, X = Cl, Br): A Family of Organically Templated Metal Halides.

Structural transformation in hybrid halides is an attractive way to modulate and improve their optical properties. Following newly reported monovalent-metal-based organic-inorganic chlorides [N(CH3)4]MCl2 (M = Ga+, In+), their bromide analogues [N(CH3)4]MBr2 (M = Ga+, In+) were subsequently synthesized. Impressively, their structure transforms from noncentrosymmetric P4̅21m space group for chlorides to centrosymmetric P42/m space group for bromides. As the halogen changes, the (MBr2)- units are partially rotated along the c-axis, which can be attributed to the difference in the anion radius and H-bond interactions between host networks and guest molecules. In addition, the tunable electronic structure and optical anisotropy in this family are also presented. This work provides a typical example for exploring new asymmetric and symmetric organic-inorganic hybrid halides as appliable photoelectric functional materials.

[Clinical and genetic analysis of a patient with rare nephronophthisis].

OBJECTIVE: To explore the genetic basis for a child with clinically suspected nephronophthisis (NPHP). METHODS: Peripheral blood samples of the patient and her parents were collected subjected to high-throughput sequencing. Sanger sequencing was used to verify the gene variants. RESULTS: The patient, a 7-year-old girl with congenital blindness, was admitted to a local hospital due to repeated vomiting for 7-8 days and then transferred to author's hospital due to renal failure. Her urine occult bloods (3+) and urine protein (1+) were abnormal. Her blood urea nitrogen and creatinine showed a significant progressive increase. Renal ultrasound showed a mild enlargement in bilateral renal, increased echogenicity, loss of corticomedullary differentiation, and the presence of cysts in both kidneys. No familial genetic history was found in the family of patient and the child was clinically diagnosed with nephronophthisis. The proband was found to harbor compound heterozygous variants of the CEP290 gene, namely c.2587-2A>T and c.2251C>T, which were inherited from her mother and father, respectively. Based on the ACMG guidelines, both variants were predicted to be pathogenic. CONCLUSION: The patient was diagnosed with NPHP type 6 due to variants of the CEP290 gene. Above finding has provided new evidence for the genotype-phenotype correlation of this disease.

Accumulation of potentially toxic elements in agricultural soil and scenario analysis of cadmium inputs by fertilization: A case study in Quzhou county.

Fertilizer application has greatly increased crop yield, however impurities in mineral or organic fertilizers, such as heavy metals, are being added to agricultural soils, which would pose a high risk for soil and crop production. 115 soil samples were collected from Quzhou, a typical agricultural county in the North China Plain, to investigate the total content of cadmium (Cd), arsenic (As), lead (Pb), nickel (Ni), copper (Cu), zinc (Zn) and chromium (Cr) in soils. The contamination levels and source apportionment of studied elements were explored by the pollution indices, multivariate statistical approaches and geostatistical analysis. The ranges of Cd, As, Pb, Ni, Cu, Zn and Cr were between 0.08 and 0.35, 5.34-15.9, 7.34-38.9, 12.9-61.3, 7.80-27.0, 31.4-154, and 17.0-50.5 mg/kg and with the mean values 0.16, 9.20, 16.0, 24.7, 17.6, 61.1, and 29.5 mg/kg, respectively. The studied area was slightly polluted mainly by Cd, and higher pollution was found in soils under vegetable crops. The application of mineral phosphate fertilizer and livestock manure were the main source of Cd and Zn, and other elements (As, Pb, Ni and Cu) might originate from soil parent materials. Scenario analyses were performed using the R programming language, based on the cadmium contents in mineral phosphate fertilizers and livestock manures. The results showed that the long-term application of phosphate fertilizers would lead to some Cd enrichment in soil without risk of substantial pollution. Compared to pure mineral fertilizers, the long-term application of blended fertilizers (30% livestock manures and 70% phosphate fertilizers) or livestock manures would incur a higher Cd pollution risk within a short period, with a maximum probability of Cd risk of 55.21%. Mitigation measurements and scientific agronomic practices should be developed to minimize the risk of potential toxic elements in agricultural soil.

[Variant analysis of CCBE1 gene in a case of Hennekam lymphangiectasia-lymphedema syndrome type 1].

OBJECTIVE: To explore the genetic etiology of a child with lymphangiectasia and lymphedema. METHODS: DNA sample of the patient was extracted and subjected to whole exome sequencing. Suspected variants were verified by Sanger sequencing. RESULTS: The patient was found to carry compound heterozygote variants (c.521G>A and c.472C>T) of the CCBE1 gene, which were respectively inherited from his parents. CONCLUSION: The compound heterozygote variants of the CCBE1 gene probably underlie the disease in this child.

An integration-free iPSC line SDQLCHi025-A from a girl with multiminicore disease carrying compound heterozygote mutations in RYR1 gene.

Peripheral blood mononuclear cells for reprogramming in this work were donated by a girl with clinically and genetically diagnosed multiminicore disease harboring compound heterozygote mutations of RYR1 gene. Induced pluripotent stem cells (iPSCs) were obtained by non-integrating episomal vectors containing OCT4, SOX2, KLF4, BCL-XL and c-MYC. The iPSC line (SDQLCHi025-A) presented pluripotent cell morphology, high mRNA levels of pluripotency markers, differentiation potential in vitro, a normal karyotype, and carrying RYR1 gene mutations.

Downregulation of orosomucoid 2 acts as a prognostic factor associated with cancer-promoting pathways in liver cancer.

BACKGROUND: Liver cancer has a high mortality and morbidity rate throughout the world. In clinical practice, the prognosis of liver cancer patients is poor, and the complex reasons contribute to treatment failures, including fibrosis, hepatitis viral infection, drug resistance and metastasis. Thus, screening novel prognostic biomarkers is of great importance for guiding liver cancer therapy. Orosomucoid genes (ORMs) encode acute phase plasma proteins, including orosomucoid 1 (ORM1) and ORM2. Previous studies showed their upregulation upon inflammation, but the specific function of ORMs has not yet been determined, especially in the development of liver cancer. AIM: To determine the expression of ORMs and their potential function in liver cancer. METHODS: Analysis of the expression of ORMs in different human tissues was performed on data from the HPA RNA-seq normal tissues project. The expression ratio of ORMs was determined using the HCCDB database, including the ratio between liver cancer and other cancers, normal liver and other normal tissues, liver cancer and adjacent normal liver tissues. Analysis of ORM expression in different cancer types was performed using The Cancer Genome Atlas and TIMER database. The expression of ORMs in liver tumor tissues and adjacent normal tissues were further confirmed using Gene Expression Omnibus data, including GSE36376 and GSE14520. The 10-year overall survival (OS), progression-free survival (PFS) and relapse-free survival (RFS) rates between high and low ORM expression groups in liver cancer patients were determined using the Kaplan-Meier plotter tool. Gene Set Enrichment Analysis (GSEA) was employed to explore the ORM2-associated signaling network. Correlations between ORM2 expression and tumor purity or the infiltration level of macrophages in liver tumor tissues were determined using the TIMER database. The correlation between ORM2 gene levels, tumor-associated macrophage (TAM) markers (including CD68 and TGFß1) and T cell immunosuppression (including CTLA4 and PD-1) in liver tumor tissues and liver GTEx was determined using the GEPIA database. RESULTS: ORM1 and ORM2 were highly expressed in normal liver and liver tumor tissues. ORM1 and ORM2 expression was significantly decreased in liver tumor tissues compared with adjacent normal tissues, and similar results were also noted in cholangiocarcinoma, esophageal carcinoma, and lung squamous cell carcinoma. Further analysis of the Gene Expression Omnibus Database also confirmed the downregulation of ORM1 and ORM2 in liver tumors. Survival analysis showed that the high ORM2 group had better survival rates in OS, PFS and RFS. ORM1 only represented better performance in PFS, but not in OS or RFS. GSEA analysis of ORM2 from The Cancer Genome Atlas liver cancer data identified that ORM2 positively associated with the G2/M checkpoint, E2F target signaling, as well as Wnt/ß-catenin and Hedgehog signaling. Moreover, apoptosis, IFN-α responses, IFN-γ responses and humoral immune responses were upregulated in the ORM2 high group. ORM2 expression was negatively correlated with the macrophage infiltration level, CD68, TGFß1, CTLA4 and PD-1 levels. CONCLUSION: The results showed that ORM1 and ORM2 were highly expressed specifically in liver tissues, whereas ORM1 and ORM2 were downregulated in liver tumor tissues. ORM2 is a better prognostic factor for liver cancer. Furthermore, ORM2 is closely associated with cancer-promoting pathways.