Methemoglobinemia caused by hemoglobin M variants is a rare inherited hemoglobinopathy characterized by lifelong cyanosis and discordant oxygen measurements. We report an 18-year-old male presenting with marked cyanosis, chocolate-brown blood, markedly reduced arterial oxygen saturation, normal arterial partial pressure of oxygen, and elevated methemoglobin levels. Complete blood count analysis on a Sysmex XN hematology analyzer revealed a global downward shift of fluorescence signals on the leukocyte differential scattergram with apparent pseudoeosinophilia, whereas parallel analysis on an ADVIA 2120 analyzer showed a normal leukocyte differential. Peripheral blood smear examination demonstrated no morphologic abnormalities. Hemoglobin electrophoresis and whole-genome sequencing identified a heterozygous HBB:c.190C > T (p.His64Tyr) mutation, confirming the diagnosis of Hb M Saskatoon-associated methemoglobinemia. This case highlights methemoglobin as an important but underrecognized source of analytical interference in fluorescence-based leukocyte differential analysis and underscores the need for integrated interpretation of scattergram features, cross-platform verification, and clinical context in automated hematology testing.

Immune checkpoint inhibitors (ICIs) have revolutionized cancer therapy by reinvigorating antitumor immunity through the blockade of inhibitory pathways such as programmed cell death protein 1 (PD-1)/programmed cell death protein ligand 1 (PD-L1) and cytotoxic T-lymphocyte-associated protein 4 (CTLA-4). Despite their remarkable clinical success, only a subset of patients derives durable benefit, whereas others exhibit primary or acquired resistance and develop immune-related adverse events (irAEs). These heterogeneous responses highlight an urgent need for robust biomarkers to predict therapeutic efficacy and for innovative combinatorial strategies to enhance clinical outcomes. Beyond their classical roles in hemostasis and thrombosis, platelets have recently emerged as pivotal modulators of tumor progression and immune regulation. Accumulating evidence indicates that platelets engage in dynamic crosstalk with tumor and immune cells, reshaping the tumor microenvironment (TME) and modulating the response to ICI therapy. Of note, platelet-associated immune checkpoint molecules (e.g., PD-L1) have shown great promise as liquid biopsy markers for patient stratification and real-time immunomonitoring. Furthermore, platelet-associated nucleic acids and traditional platelet parameters (such as platelet count and activation status) have been identified as accessible and effective biomarkers for predicting ICI responsiveness and irAEs. These platelet-derived components may also represent novel therapeutic targets to overcome resistance and potentiate ICI efficacy. Meanwhile, advances in biomaterials and genetic engineering have further enabled the development of platelet-based and platelet membrane (PM)-camouflaged delivery systems endowed with tumor-homing capacity, combinatorial drug delivery potential, and immune-responsive release properties. Collectively, these insights reposition platelets from passive participants to active regulators and versatile therapeutic platforms in cancer immunotherapy, providing a conceptual foundation for next-generation platelet-guided precision immunotherapeutic strategies.

Objective:This study aimed to evaluate the diagnostic performance of individual serum biomarkers [alpha-fetoprotein (AFP), protein induced by vitamin K absence or antagonist-II (PIVKA-II)] and composite models (GALAD, ASAP) for hepatocellular carcinoma (HCC) across two immunoassay platforms. Methods:From 2011 to 2021, 518 serum samples were selected from a liver-related disease biobank at Peking Union Medical College Hospital (Beijing, China), including 102 HCC patients, 117 with benign liver disease, 38 with cholangiocarcinoma, 96 with colorectal cancer, 65 with metastatic hepatic carcinoma, and 100 healthy controls. AFP and PIVKA-II levels were measured on both the Hotgen and Abbott ARCHITECT platforms. The GALAD and ASAP scores were calculated based on the data from each platform. Receiver operating characteristic (ROC) curve analysis and the corresponding areas under the curves (AUCs) were used to evaluate and compare the diagnostic value of the individual biomarkers and the two composite models. Results:For HCC diagnosis, AFP exhibited comparable efficacy between Hotgen (AUC: 0.821) and Abbott (AUC: 0.846), whereas PIVKA-II performed better on Abbott (AUC: 0.863) than Hotgen (AUC: 0.787). GALAD and ASAP models exhibited significantly better diagnostic performance than individual serum biomarkers on both platforms (P < 0.05): on Hotgen, both models achieved an AUC of 0.872, while on Abbott, ASAP (AUC: 0.913) was marginally superior to GALAD (AUC: 0.901, P = 0.0569). Notably, both models performed better on Abbott than Hotgen (GALAD: 0.901 vs 0.872, P = 0.0001; ASAP: 0.913 vs 0.872, P = 0.0003). Spearman correlation analysis showed moderate inter-platform correlations for AFP (r = 0.573) and PIVKA-II (r = 0.460). Bland-Altman analysis indicated poor inter-platform consistency, with mean biases of 44.32% (AFP) and -92.02% (PIVKA-II). Conclusion:GALAD and ASAP models demonstrate superior diagnostic efficacy for HCC compared to individual biomarkers, and their performance is significantly influenced by the immunoassay platform employed.

Microfluidic impedance flow cytometry has functioned as an enabling instrument in single-cell analysis, which, however, suffers from the limiting tradeoff between high sensitivity and clogging-free operation. In order to address this issue, this study presented a microfluidic impedance flow cytometer based on three-dimensional (3D) hydrodynamic focusing, in which the crossflow of conductive sample fluids and insulating sheath fluids was leveraged to centralize and restrict electric field lines to the sample fluid, thereby achieving high impedance sensitivity of single cells without the concern of channel blockage. Different from conventional impedance flow cytometry, in this study, impedance amplitude dips (rather than pulse singles) generated by single microparticles traveling through the 3D hydrodynamic focusing region were experimentally validated using microbeads. Based on the home-developed microfluidic impedance flow cytometer, high-sensitivity and clogging-free impedance profiles of three leukemia cell lines (K562, Jurkat, and HL-60) and four types of purified leukocytes (neutrophil, eosinophil, monocyte, and lymphocyte) were quantified as -8.01 ± 2.96%, -4.53 ± 1.09%, -6.36 ± 1.54%; -8.11 ± 0.84%, -7.23 ± 1.06%, -9.05 ± 2.00% and -5.68 ± 1.24%, respectively. When a recurrent neural network was adopted for cell-type classification, high classification accuracies of 93.9% for three leukemia cell lines and 87.8% for four types of purified leukocytes were achieved. This study presented a promising impedance flow cytometer that combines high sensitivity with sustainable working capabilities, potentially overcoming the limitations of conventional microfluidic impedance flow cytometry and significantly advancing its commercial development.

Leukocyte differential analysis represents a critical first-line diagnostic tool in clinical examinations. However, conventional methodologies exhibit significant limitations: Automated slide stainers demonstrate low throughput, while standard hematology analyzers lack the capability to accurately classify abnormal leukocytes. To address this, an imaging flow cytometry system was developed based on acoustic focusing to capture high-quality, label-free images of individual leukocytes, including 5 subtypes of normal cells and 3 subtypes of abnormal cells. A convolutional neural network (CNN) was employed for feature extraction and cell-type classification, enabling an 8-part leukocyte differential. Based on the developed system, high-quality label-free images of 4515 normal leukocytes and 3289 abnormal leukemia cells were obtained. Using ResNet-18, classification accuracies for 5-part normal leukocytes, 3-part abnormal leukocytes, and 8-part normal/abnormal leukocytes were quantified as 98.5 %, 97.0 %, and 97.9 %, respectively. To summarize, this label-free imaging flow cytometry enables an 8-part leukocyte differential through a combination of acoustic focusing and a deep neural network.

Cryoglobulinemic vasculitis (CryoVas) is the most common symptom of cryoglobulinemia. However, the mechanism by which cryoglobulins (CGs) induce vascular injury remains unclear. Herein, serum samples from patients with type I, II, and III cryoglobulinemia were analyzed, with healthy individuals as controls. Dynamic light scattering and electron microscopy revealed amorphous CG aggregates, with sizes increasing progressively from type I to III. Cytotoxicity and apoptosis assays demonstrated CG aggregates alone caused no direct harm to M1 macrophages or human umbilical vein endothelial cells (HUVECs). However, CGs were endocytosed by M1 macrophages, triggering secretion of TNF-α, perforin, and IL-1β, which induced HUVEC apoptosis. Type II/III CGs exhibited stronger pro-inflammatory effects than type I, correlating with their structural aggregation tendencies. These findings challenge the notion of direct CG cytotoxicity, instead implicating macrophage-mediated cytokine release as the primary driver of vascular injury in CryoVas. The study highlights immune activation-rather than intrinsic CG toxicity-as central to pathogenesis, offering new insights for therapeutic strategies targeting macrophage-derived inflammatory pathways. This mechanistic distinction between cryoglobulin types may also inform diagnostic approaches based on structural and functional profiling of CG aggregates.

In cardiovascular disease, the discovery of the proprotein convertase subtilisin/kexin type 9 (PCSK9) has undoubtedly opened a new chapter in regulating blood lipids. Since its first identification as a key regulator of low-density lipoprotein receptor (LDLR) degradation in 2003, the role of PCSK9 in cholesterol metabolism has been extensively studied. However, with further research, the pleiotropy of PCSK9 has gradually emerged, and its impact extends far beyond cholesterol metabolism in the liver. The purpose of this review is to systematically explore the pleiotropy of PCSK9, extending from its important role in lipid regulation to its extensive effects in extrahepatic tissues, and to reveal its potential role in cardiovascular health, nervous system function, and tumor biology. By integrating the latest research findings, this paper summarizes the complex mechanisms of action of PCSK9 in different biological processes and explores its potential and challenges as a therapeutic target.

This study presented a high-sensitivity and clogging-free microfluidic impedance flow cytometer enabled by virtual constriction microchannel. The new structure of the virtual constriction microchannel relied on crossflowing conductive cell solutions and insulated sheath fluids, leading to high impedance sensitivities without channel blockage. Large impedance variations (similar to 1%) of a variety of microparticles were captured smoothly by this cytometer, and based on deep neural networks, high accuracies of 94.4% and 87.8% in classifying leukemia cell lines and 4-part leukocytes were obtained.

This paper presented a high-quality imaging flow cytometry based on acoustic focusing with demonstrative applications in label-free leukocyte differential coupled with deep neural network. Novelties of this study lied in 1) capture of high-quality cell images due to acoustic radiation forces, where all cells travelled along the standing wave node without deviation, enabling similar to 100% capture of cells without losing focuses; 2) high classification accuracy due to deep neural network, where the best performance of leukocyte differential (99.4% at 5-part normal/abnormal leukocytes) was reported among label-free impedance and/or imaging flow cytometry.

(1) Background: Methylobacterium radiotolerans (M. radiotolerans) is a fastidious, aerobic, Gram-negative bacillus primarily found in environmental sources such as soil and sewage, with rare clinical isolation. Its identification remains challenging due to poor growth with conventional culture methods. (2) Case presentation: A 42-year-old male patient with early T-cell precursor acute lymphoblastic leukemia (ETP-ALL) presented with M. radiotolerans bacteremia during hospitalization. The organism was successfully isolated from peripheral blood using the Myco/F Lytic culture vial (Becton, Dickinson and Company, Lincoln, MT, USA). Comparative analysis demonstrated markedly superior growth of M. radiotolerans in Myco/F Lytic culture vials compared with Plus Aerobic/F Lytic and Lytic/10 Anaerobic/F culture vials (Becton, Dickinson and Company, Lincoln, MT, USA). Antimicrobial susceptibility testing, performed with the epsilometer test (E-test) and Bauer–Kirby disk diffusion (BK) method, guided the selection of an appropriate therapeutic regimen. The patient’s infection was ultimately controlled following targeted antimicrobial therapy. (3) Conclusions: M. radiotolerans demonstrates a distinct growth preference for the Myco/F Lytic culture medium. This observation highlights the importance of considering alternative culture media in cases of rare or fastidious bacterial infections that cannot be reliably detected using conventional Plus Aerobic/F Lytic or Lytic/10 Anaerobic/F culture vials, which are typically employed for clinical isolation of aerobic and anaerobic bacteria.

DNAJB6, a major member of the DNAJ/HSP40 family, plays an important role in tumor development. We explored the effect of DNAJB6 expression on the prognosis of patients and its biological role in lung adenocarcinoma (LUAD). mRNA and clinical data were obtained from The Cancer Genome Atlas (TCGA). Enriched pathways were determined by the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. A nomogram incorporating DNAJB6 and three clinical features was constructed to predict the survival rate. DNAJB6 expression and function in LUAD were explored using immunohistochemistry, Western blotting, proliferation, cell cycle analysis, RNA sequencing, and xenograft tumor assays. DNAJB6 mRNA levels were elevated in the LUAD-TCGA dataset. DNAJB6 protein levels were higher in LUAD tumor tissues than in normal tissues. A high DNAJB6 level was an independent risk factor for poor prognosis in patients with LUAD. The proportion of tumor-infiltrating immune cells significantly differed between high and low DNAJB6 expression. DNAJB6 was associated with cell cycle pathways; therefore, its knockdown induced G2/M cell cycle arrest and inhibited LUAD cell proliferation. This is the first report of the DNAJB6 requirement for LUAD cell proliferation and its potentially crucial role in LUAD prognosis.

Objective:Serum pepsinogen (PG) is a good indicator of atrophic changes in the gastric mucosa. Gastric mucosal atrophy is a high-risk factor for gastric cancer. Serological testing for PG combined with endoscopy can help to improve gastric cancer screening. In this study, we established the reference ranges of serum PG-I, PG-II, and the PG-I/II ratio (PGR) in the Chinese population by chemiluminescence immunoassay (CLIA) and enzyme-linked immunosorbent assay (ELISA). Besides, in the real world, doctors are often confused by the results of different testing platforms. Thus, a comparison of methods CLIA and ELISA was performed. Methods:2904 individuals were enrolled from six regions in China as part of the Chinese Adult Digestive Diseases Surveillance (2016) program. The individuals completed questionnaires and volunteered to undergo examinations, including gastroscopy, urea breath test, abdominal ultrasound examination and routine serologic tests. Serum was collected to measure PGs (including PG-I, PG-II and PGR) by CLIA and ELISA. Participants who were found obvious abnormalities or absent from the examinations were excluded. Ultimately, 747 healthy individuals were enrolled in this study. The Kolmogorov-Smirnov test was used to assess the distribution of variables. The Kruskal-Wallis H or Mann-Whitney U-tests were used to compare different sex, age, and geographical groups. The 95% reference ranges of PGs obtained by the two methods were established according to document CLSI-EP28-A3, with covariates of sex, age, and region. Spearman correlation analysis, linear regression analysis and allowable total error (ATE) zone analysis were utilized for comparing the two methods. Results:On overall, the 95% reference ranges of PG-I, PG-II, and PGR measured by CLIA were 23.00-110.64 ng/mL, 2.50-19.13 ng/mL, and 3.87-13.30, respectively. Meanwhile, the reference ranges of PG-I, PG-II, and PGR measured by ELISA were 36.93-205.06 ng/mL, 1.65-17.96 ng/mL, and 7.50-33.60, respectively. Both PG-I and PG-II levels measured by the two platforms were found to be influenced by sex and age. PGR measured by CLIA was influenced by age but not by sex, while PGR measured by ELISA was not affected by either age or sex. Regional factors did not significantly impact the PG results, except for PG-I detected by ELISA. Ultimately, reference ranges for PGs were established based on age and sex stratification. Additionally, the Spearman correlation analysis revealed that the correlation coefficients for PG-I, PG-II, and PGR detected by the two methods were 0.899, 0.887, and 0.777, respectively, indicating a strong correlation between the two methods. The regression equation for the PG levels detected by two methods was obtained through linear regression analysis. The ATE analysis provided a visual depiction of the consistency between the two methods, clearly indicating the poor agreement between them. Conclusion:This study established the reference ranges of PGs by strict and intact enrollment standard. In addition, the results indicated a strong linear relationship between the two methods, yet with a clear bias, which was valuable for laboratory interpretation.

Waldenstr & ouml;m macroglobulinemia (WM) is a relatively rare hematological malignancy characterized by serum monoclonal IgM gammopathy and bone marrow infiltration of lymphoma cells (small B lymphocytes, plasmacytoid lymphocytes, or plasma cells). Elevated CA125 is most seen in ovarian cancer or some benign diseases such as pelvic inflammatory disease and endometriosis. No cases of WM combined with elevated CA125 have been reported so far. Here, we report two rare cases of WM with abnormally high CA125 at the onset of illness. Patient 1 had a nine-year history of pulmonary shadow with a moderately increased CA125 level. Subsequently, she was diagnosed with WM-related lung involvement by biopsy. Patient 2 presented with WM manifestation and a significantly elevated CA125 level of unknown significance. Based on bone marrow smear results and serum IgM levels, the diagnosis of WM was established in both patients. After rigorous physical examination, imaging screening, and pathological biopsy, any underlying disease associated with elevated CA125 in both patients was excluded. CA125 and IgM levels decreased with effective treatment for WM, suggesting that abnormally elevated CA125 was related to the progression of macroglobulinemia. Suspicious WM patients with elevated serum CA125 of unknown significance need to be alert to a special manifestation of macroglobulinemia. More clinical concern is needed. At the same time, the clinician could monitor the patient's serum CA125 level changes to assist in the judgment of the efficacy of the original disease. This report extends the understanding of WM and the application of CA125.

To facilitate adeno-associated virus (AAV)-mediated gene therapy in China, we conducted a study on the distribution of AAV-neutralizing antibodies (NAbs) in healthy subjects and in patients with Duchenne muscular dystrophy (DMD)/Becker muscular dystrophy (BMD). A total of 352 healthy adult controls (ACs) from a national multicenter study, 100 schoolchild controls (SCs), and 281 patients with DMD/BMD from Peking Union Medical College Hospital were enrolled in this study. Cell-based inhibition assays were applied, and serum samples demonstrating 50% inhibition of infection were considered positive. The seroprevalence of AAV2 and AAV9 NAbs among the 733 participants was 86.1% and 56.3%, respectively. The AAV2 NAbs and AAV9 NAbs positivity rates in the AC, SC, and DMD/BMD groups were 97.4%/86.6%, 100.0%/17.0%, and 66.9%/32.4%, respectively. The seroprevalence of AAV NAbs gradually increased with age, especially in AAV9 NAbs. Females tended to have higher positivity rate than males. Over 85% of ACs had overlapping AAV9 and AAV2 infection. However, being positive for only AAV2 NAbs in the SC group was common, and 30.6% of patients with DMD/BMD were negative for both AAV2 and AAV9 NAbs. Our findings reveal that a significant proportion of patients with DMD/BMD were negative for AAV2 and AAV9 NAbs, which is the population that is most amenable to being treated with gene therapy.

The differential of leukocytes functions as the first indicator in clinical examinations. However, microscopic examinations suffered from key limitations of low throughputs in classifying leukocytes while commercially available hematology analyzers failed to provide quantitative accuracies in leukocyte differentials. A home-developed imaging and impedance flow cytometry of microfluidics was used to capture fluorescent images and impedance variations of single cells traveling through constrictional microchannels. Convolutional and recurrent neural networks were adopted for data processing and feature extractions, which were then fused by a support vector machine to realize the four-part differential of leukocytes. The classification accuracies of the four-part leukocyte differential were quantified as 95.4% based on fluorescent images plus the convolutional neural network, 90.3% based on impedance variations plus the recurrent neural network, and 99.3% on the basis of fluorescent images, impedance variations, and deep neural networks. Based on single-cell fluorescent imaging and impedance variations coupled with deep neural networks, the four-part leukocyte differential can be realized with almost 100% accuracy.

Celiac disease (CD), a gluten‐related disease, is a multi‐system rare disorder mainly involving the gastrointestinal tract. The clinical signs of CD are exceedingly heterogeneous, which increases the difficulty of clinical differential diagnosis. Neurological manifestations are one of the non‐classical CD symptoms. As some patients present only neurological symptoms at early stages, the diagnosis of CD is always delayed. Correct diagnosis and management could decrease patient morbidity and deaths. A 32‐year‐old male was admitted to the hospital due to progressive muscle atrophy of both lower limbs and lumbar stiffness. Based on positive gluten‐sensitive enteropathy autoantibody profiles and gastroscopy foundation, the diagnosis of CD was established. The patient was instructed to gluten‐free diet. The antibody titer of gluten‐sensitive enteropathy autoantibodies decreased, and the patient's symptoms alleviated. We emphasize the importance of CD screening in patients with neurological disorders of unknown aetiology.

To investigate the prevalence of gastroesophageal reflux disease (GERD), reflux esophagitis (RE), digestive ulcer gastric ulcer (GU), duodenal ulcer (DU), and Helicobacter pylori infection in Chinese adults aged 18-64 years and their associated factors, a community-based cross-sectional study using a stratified multistage sampling method was conducted. A standardized questionnaire survey, the 13C-urea breath test, and gastroscopy were performed. Weighted methods were used to estimate the prevalence of diseases or infection mentioned above and their risk factors. Finally, 27,637 participants aged 18-64 years were enrolled from 2017 to 2018. The prevalence (95% confidence interval) of GERD, RE, GU, DU, and H. pylori infection was estimated to be 10.5% (7.8%-14.2%), 5.4% (3.9%-7.3%), 2.5% (1.7%-3.7%), 4.5% (3.6%-5.4%), and 41.5% (36.7%-46.4%), respectively. The fraction of H. pylori infection reached 58.6% and 61.1% among the GU and DU patients, respectively. Weighted multivariable logistic regression models showed that GERD, RE, and GU shared the common risk factors of age and obesity. Dose-response relationships were observed between smoking and all four diseases, as well as alcohol consumption and GERD and H. pylori infection. Northwest China had the highest prevalence of GERD (23.9%), RE (8.7%), GU (7.8%), DU (7.3%), and H. pylori infection (63.6%); however, the southwest region had the highest prevalence of GU but the lowest of DU, RE, and H. pylori infection. Non-steroidal anti-inflammatory drugs were positively associated with GERD risk. On the contrary, a reduced risk of GU was observed among H. pylori-infected patients taking this drug. In summary, the prevalence of GERD, RE, and H. pylori infection still appears high in China. H. pylori infection eradication remains the priority to reduce the burden of peptic ulcer disease. The aging population, high prevalence of overweight or obesity, smoking, and drinking in China could explain the high burden of these diseases, thus suggesting the targeted preventive measures for upper gastrointestinal diseases in the future.

This study presented a microfluidic impedance flow cytometry based on three-dimensional hydrodynamic focusing, in which the sample fluid (e.g., cells or microbeads) with high conductivity was focused to the center of the detection microchannel by the sheath fluid with low conductivity. As a result, electric field lines were centralized and restricted to the sample fluid, leading to large impedance variations caused by travelling single cells in the detection microchannel. Optimal parameters of the three-dimensional hydrodynamic focusing structure were determined by finite element simulations and validated by experimental observations. Three-frequency impedance profiles of individual microbeads travelling through the three-dimensional hydrodynamic focusing structure under a variety of focusing circumstances were captured and compared, validating the functionality of the microfluidic impedance flow cytometry of this study.

Long noncoding RNAs (lncRNAs) have emerged as a new family of master regulators of cancer. The HOXA transcript antisense RNA myeloid-specific 1 (HOTAIRM1) located in the HOXA gene cluster was first discovered in acute promyelocytic leukemia. Increasing evidence have showed that HOTAIRM1 is involved in diverse cancer. It regulates tumor proliferation, invasion, and migration via various mechanisms and is related to the clinicopathological characteristics of tumor patients. This article reviews its expression, function, and molecular mechanism in various malignant tumors and future challenges for the diagnosis and treatment of cancers through HOTAIRM1.

Abstract Besides medullary thyroid cancer, a latent and relatively mild increase of serum procalcitonin (PCT) has been found in neuroendocrine neoplasms (NENs). Here we are aiming to supply more information about NENs related PCT elevation, in order to broaden the clinical experiences about diagnosis and treatment of shock in tumor patients. we reported an advanced pancreatic neuroendocrine carcinoma (pNEC) with liver and lung metastasis, in which a rare pseudo-sepsis shock with extremely high serum PCT level (exceeding 100 ng/ml) had been demonstrated. A series of screening tests to exclude bacterial infections, including blood culture, urine culture and even metagenomic NGS (mNGS), had been performed. Given negative evidence of bacterial infection and useless broad-spectrum antibiotics treatment, steroid was used to relieve the serious inflammation and its related shock. The patient’s condition was improved and discharged. In summary, despite the ubiquitous use of PCT used in bacterial infection and sepsis shock, pNEC could cause the high level of serum PCT and even result in severe inflammation accompanied by shock. As for diagnosis and treatment strategies, pNEC should be regarded as one of rare differential diagnosis when experimental antibiotics are not working. The potential mechanism of PCT elevation and its role in prognosis of pNEC still needed to be further studied.