{"data":{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:40225293", "kind": "science-record", "revision": 4, "content_hash": "sha256:4d03c6f878e590bc32dc674a90a92d134d3aedfd872eb67b667b0111fec0d176", "created_at": "2026-11-01T20:30:23Z", "updated_at": "2026-11-01T22:56:21Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "Identification and Verification of Key Genes Associated with Temozolomide Resistance in Glioblastoma Based on Comprehensive Bioinformatics Analysis.", "doi": "10.30498/ijb.2024.448826.3892", "pmid": "40225293", "authors": ["Jun Hu", "Jingyan Yang", "Na Hu", "Zongting Shi", "Tiemin Hu", "Baohong Mi", "Hong Wang", "Weiheng Chen"], "journal": "Iranian journal of biotechnology", "year": 2024, "abstract": "Background: Glioblastoma (GBM) is the most aggressive form of brain cancer, with poor prognosis despite treatments like temozolomide (TMZ). Resistance to TMZ is a significant clinical challenge, and understanding the genes involved is crucial for developing new therapies and prognostic markers. This study aims to identify key genes associated with TMZ resistance in GBM, which could serve as valuable biomarkers for predicting patient outcomes and potential targets for treatment.\nObjectives: This study aimed to identify genes involved in TMZ resistance in GBM and to assess the value of these genes in GBM treatment and prognosis evaluation.\nMaterials and Methods: Bioinformatics analysis of Gene Expression Omnibus (GEO) datasets (GSE113510 and GSE199689) and The Chinese Glioblastoma Genome Atlas (CGGA) database was performed to identify differentially expressed genes (DEGs) between GBM cell lines with and without TMZ resistance. Subsequently, the key modules associated with GBM patient prognosis were identified by weighted gene coexpression network analysis (WGCNA). Furthermore, hub genes related to TMZ resistance were accurately screened and confirmed using three machine learning algorithms. In addition, immune cell infiltration analysis, TF-miRNA coregulatory network analysis, drug sensitivity prediction, and gene set enrichment analysis (GSEA) were also performed for temozolomide resistance-specific genes. Finally, the expression levels of key genes were validated in our constructed TMZ-resistant cell lines by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting (WB).\nResults: Integrated analysis of the GEO and CGGA datasets revealed 769 differentially expressed genes (DEGs), comprising 350 downregulated and 419 upregulated genes, between GBM patients and normal controls. Among these DEGs, three key genes, namely, PITX1, TNFRSF11B, and IGFBP2, exhibited significant differences in expression between groups and were prioritized via machine learning algorithms. The expression levels of these genes were found to be closely related to adverse clinical features and immune cell infiltration levels in GBM patients. These genes were also found to participate in several biological pathways and processes. RT‒qPCR and WB confirmed the differential expression of these genes in vitro, indicating that they play vital roles in GBM patients with TMZ resistance.\nConclusions: PITX1, TNFRSF11B, and IGFBP2 are key genes associated with the prognosis of GBM patients with TMZ resistance. The differential expression of these genes correlates with adverse outcomes in GBM patients, suggesting that they are valuable biomarkers for predicting patient prognosis and that they could serve as diagnostic biomarkers or treatment targets.", "claims": [{"id": "claim-method", "statement": "After normalization and comparison of gene expression levels between the TMZ-sensitive and TMZ-resistant groups, DEGs were screened using the R package “limma” with the cutoff criteria |log2-fold change| >1 and adjusted p value < 0.05.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:40225293", "locator": "sections[5] \"3.2. Identification of DEGs\" paragraph 2", "quote": "After normalization and comparison of gene expression levels between the TMZ-sensitive and TMZ-resistant groups, DEGs were screened using the R package “limma” with the cutoff criteria |log2-fold change| >1 and adjusted p value < 0.05."}], "state": "complete"}, {"id": "claim-data-access", "statement": "The study’s datasets are accessible online via the following GEO accession numbers: GSE113510 and GSE199689, available at NCBI’s Gene Expression Omnibus (https://www.ncbi.nlm.nih.gov/geo).", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:40225293", "locator": "sections[28] \"ack\" paragraph 1", "quote": "The study’s datasets are accessible online via the following GEO accession numbers: GSE113510 and GSE199689, available at NCBI’s Gene Expression Omnibus (https://www.ncbi.nlm.nih.gov/geo)."}], "state": "complete"}, {"id": "claim-target-result", "statement": "Integrated analysis of the GEO and CGGA datasets revealed 769 differentially expressed genes (DEGs), comprising 350 downregulated and 419 upregulated genes, between GBM patients and normal controls.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:40225293", "locator": "sections[0] \"Abstract\" paragraph 4", "quote": "Integrated analysis of the GEO and CGGA datasets revealed 769 differentially expressed genes (DEGs), comprising 350 downregulated and 419 upregulated genes, between GBM patients and normal controls."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "Integrated analysis of the GEO and CGGA datasets revealed 769 differentially expressed genes (DEGs), comprising 350 downregulated and 419 upregulated genes, between GBM patients and normal controls.", "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:40225293"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. This lane did not download the dataset or read its licence.", "duration_hours": null, "parameters": {}, "inputs": ["data:geo:GSE113510"], "outputs": ["the source data with its repository provenance"], "instruments": [], "reagents": []}, {"id": "run_analysis", "order": 3, "name": "Run the published analysis", "kind": "analysis", "state": "complete", "blocked_on": null, "description": "Apply the paper's statistical method to the resolved series and compare the result with the published target.", "duration_hours": null, "parameters": {"fold_change": "e cutoff criteria |log2-fold change| >1 and adjusted p valu"}, "inputs": ["data:geo:GSE113510"], "outputs": ["the replicated result and its comparison"], "instruments": [], "reagents": []}], "materials": [], "instruments": [], "reagents": [], "conditions": {}, "gaps": []}, "data": {"state": "partial", "assets": [{"id": "source:pmid:40225293", "role": "manuscript", "format": "xml", "media_type": "application/xml", "uri": 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the following GEO accession numbers: GSE113510 and GSE199689, available at NCBI’s Gene Expression Omnibus (https://www.ncbi.nlm.nih.gov/geo).", "structure": {"repository": "GEO", "access_route": "GEO accession URL", "access_statement_verbatim": "The study’s datasets are accessible online via the following GEO accession numbers: GSE113510 and GSE199689, available at NCBI’s Gene Expression Omnibus (https://www.ncbi.nlm.nih.gov/geo).", "access_statement_kind": "data_availability_statement", "sample_accessions": ["GSM3108207", "GSM3108208", "GSM3108209", "GSM3108210", "GSM3108211", "GSM3108212"], "sample_count": 6, "group_sizes": null, "group_sizes_basis": null, "platform": "21047", "taxon": "Homo sapiens", "title": "The different expression of long non-coding RNAs and messenger RNAs in temozolomide-resistant glioblastoma cells and parental glioblastoma cells", "resolution_checked_at": "2026-11-01T18:46:34Z", "resolution_check": "identifier_resolution_only_no_download_no_licence_read", "downloaded": false, "licence_read": false}}, {"id": "data:geo:GSE199689", "role": "raw", "format": "geo-series", "media_type": "application/vnd.ncbi.geo-series+json", "uri": "https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE199689", "sha256": null, "bytes": null, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "GSE199689", "version": null, "note": "The study’s datasets are accessible online via the following GEO accession numbers: GSE113510 and GSE199689, available at NCBI’s Gene Expression Omnibus (https://www.ncbi.nlm.nih.gov/geo).", "structure": {"repository": "GEO", "access_route": "GEO accession URL", "access_statement_verbatim": "The study’s datasets are accessible online via the following GEO accession numbers: GSE113510 and GSE199689, available at NCBI’s Gene Expression Omnibus (https://www.ncbi.nlm.nih.gov/geo).", "access_statement_kind": "data_availability_statement", "sample_accessions": ["GSM5981753", 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No dataset was downloaded and no licence was read."}}, "can_run": {"dsl_operations": ["normalise_counts", "differential_expression_fit", "fdr_bh"], "assets": ["source:pmid:40225293", "data:geo:GSE113510"], "steps": ["run_analysis"]}, "must_build": {"dsl_operations": [], "converters": [], "readers": [], "capabilities": [], "data_gaps": ["data:geo:GSE113510"]}, "gaps": []}, "review": {"state": "partial", "machine_critique": {"kind": "extraction_review", "reader": {"model": "MiniMax-M3.1-Flash", "provider": "minimax", "provider_family": "minimax", "read_at": "2026-11-01T19:50:03Z", "independence": "single_reader"}, "evidence_check": {"method": "verbatim", "availability": "verbatim"}, "calls": 5, "usage": {"prompt_tokens": 10971, "completion_tokens": 1568, "cached_tokens": 264}}, "human_critique": null, "stages": [{"id": "extraction", "state": "complete", "reader": "MiniMax-M3.1-Flash", "provider": "minimax"}], "gaps": ["review.independent_reader", "review.machine_critique", "review.human_critique"]}, "execution": {"state": "absent", "status": "not_run", "mode": null, "runs": [], "gaps": ["execution.not_run"]}, "provenance": {"entities": [{"id": "pmid:40225293", "type": "paper", "label": "pmid:40225293"}, {"id": "source:pmid:40225293", "type": "manuscript", "label": "PMID 40225293 full text"}, {"id": "data:geo:GSE113510", "type": "dataset", "label": "GEO GSE113510"}], "activities": [{"id": "activity-reading-sci-2026-11-01", "type": "classification", "used": [{"id": "pmid:40225293", "type": "paper", "label": "pmid:40225293"}, {"id": "source:pmid:40225293", "type": "manuscript", "label": "PMID 40225293 full text"}, {"id": "data:geo:GSE113510", "type": "dataset", "label": "GEO GSE113510"}], "agent": "agent:corpus-assay-engine"}, {"id": "activity-read-40225293", "type": "extraction", "used": ["source:pmid:40225293"], "agent": "agent:minimax-reader"}, {"id": "activity-resolve-GSE113510", "type": "accession_resolution", "used": ["data:geo:GSE113510"], "agent": "agent:corpus-assay-engine"}], "agents": [{"id": "agent:corpus-assay-engine", "type": "software", "label": "Corpus assay engine"}, {"id": "agent:minimax-reader", "type": "software", "label": "MiniMax literature reader"}], "source_run_id": "reading-sci-2026-11-01", "dsl_catalogue_fingerprint": "343fb30756ba6ec3", "capability_table_fingerprint": "2cb3e42f047d2ecf", "snapshot_id": "ba34b49490d9bfaf"}, "lineage": {"parent_id": "pmid:40225293", "parent_hash": "sha256:3eff6fe652b5748405b0a8e55c6b7b917818039d9636f2d7773339558f9bdee8", "fork_of": null, "generation": 3}, "gaps": ["asset:data:geo:GSE113510 licence not read", "asset:data:geo:GSE113510 not downloaded", "asset:data:geo:GSE199689 licence not read", "asset:data:geo:GSE199689 not downloaded", "execution.not_run", "review.human_critique", "review.independent_reader", "review.machine_critique"], "assays": [{"role": "data_generation", "definition_id": "qpcr", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["time quantitative polymerase chain reaction (RT-qPCR) and Western blotting (WB). Results: Integra", "e quantitative polymerase chain reaction (RT-qPCR) and Western blotting (WB). Results: Integra", "everal biological pathways and processes. RT‒qPCR and WB confirmed the differential expression"], "order_basis": "paper_text_order:1689", "id": "assay:1"}, {"role": "data_generation", "definition_id": "western_blot", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["tive polymerase chain reaction (RT-qPCR) and Western blotting (WB). 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Evidence bands order textual support; method agreement intervals are null (n=0), never probabilities. Work-type validation does not validate methods.","Operation coverage describes only explicitly named methods. It does not prove a complete protocol, compatible parameters, successful compilation or re-execution. Unrecognised or ambiguous methods require review.","Data reachability is a textual accession or public-data statement. No dataset was downloaded or checked for access, completeness or licence.","Capabilities are planning states seeded from the dated equipment register. No bench ownership, completed quote or partner booking is inferred. Procurement counts are dependencies, not guaranteed replications or financial returns.","Tier and blocker counts are deterministic assessments of this corpus, not measured agreement. R3 is reserved for procurement; unresolved methods and data require human work (R4). The most restrictive axis wins, with capability, method, data as tie-breaks. 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At the evidence floor the work type comes from title framing and a fixed prior, not from anything in the paper, so the published work-type split should be read as a rough triage signal rather than a measured distribution. The impact is bounded but not removed: since 2026-10-23 the engine excludes floor-confidence records from the shortlist entirely and publishes them separately as held out, so the recommendation is unaffected even though the census is.","status":"unverified","severity":"high"},{"code":"runtime_is_modelled","basis":["Runtime is summed from published per-assay hour figures. Timing awaits the first executed replication.","4,407 of 5,529 records (79.7%) model to 0.00 hours because no assay was matched, which the ranking then reads as \"cheapest\".","0 of 10 shortlisted records carry a 0.00-hour estimate."],"claim":"Estimated replication runtime","gloss":"The hour figures are estimates built from assay mentions; timing awaits the first executed replication.","detail":"The hour figures are a cost model derived from the bench steps implied by each matched assay family. The first executed replication will supply measured timing. Records without an assay match currently receive a zero-hour model value.","status":"unverified","severity":"high"},{"code":"signal_score_without_citations","basis":["5,529 of 5,529 records (100.0%) have no citation count from the source metadata."],"claim":"Signal score","gloss":"The signal score uses publication-type weight because the mirror supplied zero citation counts.","detail":"The source supplied no citation counts for any record in this run, so the signal score is publication-type weight only. The citation term of the ranking is inert here, and the shortlist is ordered by publication type alone.","status":"unverified","severity":"high"},{"code":"abstract_absent","basis":["1,667 of 5,529 records (30.2%) carry no abstract at all."],"claim":"Evidence base for classification","gloss":"Some records contain a title and publication type only, defining the available classification evidence.","detail":"Where the mirror contains a title and publication types only, those fields define the classifier evidence. A methods section is required for fuller automated and human review.","status":"unverified","severity":"medium"},{"code":"classifier_agreement_measured","basis":["Published two-way decision agreement 77.7% (310 of 399 decidable records) from a stratified hand-check of 438 real records.","Three-way diagnostic agreement 73.9% (288 of 390 decidable records).","data_only agreement 75.2% (100 of 133 decidable).","48 record(s) could not be decided from title and abstract and were excluded from the denominator.","56 analysis-only records were published under another work type, so the data_only share is not an upper bound."],"claim":"Published automatable decision","gloss":"This figure has been checked against a human reading the same papers, so we know how often it is right.","detail":"The published automatable / not-automatable distribution comes from a rule-based classifier that was hand-checked against a fixed ground-truth definition. The agreement rates below are measured, not estimated. The three-way work type remains a diagnostic and is reported separately.","status":"measured","severity":"high"},{"code":"llm_comparison_not_measured","basis":["rule_based_v1+lex99e1195c7d6f produced 5,529 of 5,529 classifications.","Measured on 438 labelled records; these are RULE-BASED figures, not model results."],"claim":"Rule-based versus LLM agreement","gloss":"Every number here is the rule engine’s own. Model comparison begins when a working API credential is connected and measured.","detail":"The published two-way decision agreed with a human reading on 310 of 399 decidable records (77.7%). The diagnostic three-way label agreed on 288 of 390 decidable records (73.9%). Every percentage in this run belongs to the rule engine. Model comparison begins when a working API credential is connected and measured.","status":"not_measured","severity":"high"},{"code":"confidence_is_a_band","basis":["low band: 135 of 210 hand-judged records were correct, 64.3% [57.6, 70.5] (95% CI).","medium band: 80 of 104 hand-judged records were correct, 76.9% [68.0, 84.0] (95% CI).","high band: 73 of 76 hand-judged records were correct, 96.0% [89.0, 98.7] (95% CI).","A fitted calibrated probability was built, measured and withdrawn: it overstated held-out accuracy and reorders nothing."],"claim":"Confidence band and evidence score","gloss":"The confidence band carries a measured accuracy interval; 0.85 represents relative rank strength.","detail":"Confidence on this run is a BAND plus its measured accuracy interval. The number stored alongside each record is a monotone evidence score that orders records; 0.85 represents relative rank strength. A fitted probability was withdrawn after it overstated held-out accuracy. Threshold on the band.","status":"measured","severity":"medium"},{"code":"three_way_split_is_diagnostic","basis":["The corpus is published at the two-way decision: automatable or not-automatable. The three-way split (data_only / physical / mixed) is published as a diagnostic with its measured reliability attached, not as a claim, because the rule engine's `mixed` call cannot be defended.","The engine called 22 record(s) mixed; 10 matched the human judgement (45.5%, 95% CI 26.9-65.3).","The engine's `mixed` call is right on 10/22 hand-judged records (45.5%, 95% CI 26.9%-65.3%); a constant `physical` guess scores 36.4%.","The engine's `mixed` call does not reliably beat a constant `physical` guess on the hand-judged records it labels mixed.","The sign of that difference flips between two independent samples, so the three-way refinement is not demonstrably informative.","The mixed stratum contributes a disproportionate share of errors relative to its size.","At title-only depth the physical/mixed split collapses: a title says that work was done, not whether it was also analysed."],"claim":"The published decision is two-way","gloss":"The three-way split is shown for diagnosis only. The published split is two ways.","detail":"Count and filter the corpus on automatable versus not-automatable. The data_only / physical / mixed split remains an internal engine diagnostic because the mixed call has lower measured reliability.","status":"measured","severity":"medium"},{"code":"automatable_flag_unproven","basis":["6 of 29 assay families are marked automatable now: meta_analysis, machine_learning, survival_analysis, pca, molecular_docking, differential_expression.","The flag is a rule over assay family and work type; validation begins with the first executed replication.","It inherits the 75.2% data_only agreement rate, because the flag requires the record to be analysis-only."],"claim":"Automatable now","gloss":"This flag marks assay families the rules classify as data-only. The first executed replication will validate it.","detail":"This flag marks families the rules consider replicable from data alone. Its validity depends on the analysis-only classification and will be measured through an executed replication.","status":"unverified","severity":"medium"}],"confidence_policy":"band_with_measured_interval","confidence_contract":{"policy":"band_with_measured_interval","summary":"Confidence is reported as a BAND with a measured accuracy interval. The raw evidence score supplies monotone ordering. A fitted point estimate was withdrawn after it overstated held-out accuracy.","evidence_score":"A monotone evidence score in [0,1] built from cue mass, axis separation and text coverage. Use it to order records. Interpret 0.85 as relative rank strength.","confidence_band":"low | medium | high. Consumers threshold on this value.","band_accuracy":"Wilson 95% interval on the measured agreement of the hand-judged records in this band.","calibrated_confidence":"WITHDRAWN. Reintroduction requires held-out evidence demonstrating reliable estimates across every bin."},"band_accuracy":{"low":{"n":210,"low":0.576,"band":"low","high":0.7046,"agree":135,"accuracy":0.6429},"high":{"n":76,"low":0.8903,"band":"high","high":0.9865,"agree":73,"accuracy":0.9605},"medium":{"n":104,"low":0.6795,"band":"medium","high":0.8397,"agree":80,"accuracy":0.7692}},"provider":"rule_based_v1+lex99e1195c7d6f","lexicon_id":"99e1195c7d6f","contract_published_at":"2026-10-23T12:25:29.022232+00:00","confidence_is_band":true,"confidence_is_probability":false,"decision_taxonomy":"two_way","decidable_fraction_two_way":0.8854,"decidable_fraction_two_way_low":0.8565,"decidable_fraction_two_way_high":0.9143,"decidable_fraction_three_way":0.8586,"decidable_fraction_three_way_low":0.8287,"decidable_fraction_three_way_high":0.8884,"decidable_population":5529,"validation_sample_size":438,"three_way_cost_fraction":0.0268,"three_way_cost_records":148,"decision_counts":{"automatable":230,"not_automatable":5299},"decision_split":{"automatable":{"count":230,"share":0.04159884246699222},"not_automatable":{"count":5299,"share":0.9584011575330078}},"diagnostic_work_type_counts":{"mixed":24,"physical":5275,"data_only":230}}}