[Analysis of microsatellite instability in endometrial cancer: The significance of minimal microsatellite shift].

OBJECTIVE: To analyze the differences and characteristics of microsatellite instability (MSI) in endometrial cancer (EMC), by using colorectal cancer (CRC) as control. METHODS: In the study, 228 cases of EMC were collected. For comparative analysis, 770 cases of CRC were collected. Mismatch repair (MMR) expression was detected by immunohistochemistry (IHC), and microsatellite instability (MSI) was analyzed by PCR and capillary electrophoresis fragment analysis (MSI-PCR). MSI-PCR was detected using five mononucleotide repeat markers: BAT-25, BAT-26, NR-21, NR-24, and MONO-27. RESULTS: In EMC, we found 27.19% (62/228) of deficient mismatch repair (dMMR) using IHC, significantly higher than CRC (7.79%, 60/770). Meanwhile, subclonal expression of MMR protein was found in 4 cases of dMMR-EMC and 2 cases of dMMR-CRC. According to the criteria of major micro-satellite shift, we found 16.23% (37/228) of MSI-high (MSI-H), 2.63% (6/228) of MSI-low (MSI-L), and 81.14% (185/228) of microsatellite stability (MSS) in EMC using MSI-PCR. The discor-dance rate between MMR-IHC and MSI-PCR in EMC was 11.84% (27/228). In CRC, we found 8.05% (62/770) of MSI-H, 0.13% (1/770) of MSI-L, and 91.82% (707/770) of MSS. The discordance rate between MMR-IHC and MSI-PCR in CRC was only 0.52% (4/770). However, according to the criteria of minimal microsatellite shift, 12 cases of EMC showed minimal microsatellite shift including 8 cases of dMMR/MSS and 4 cases of dMMR/MSI-L and these cases were ultimately evaluated as dMMR/MSI-H. Then, 21.49% (49/228) of EMC showed MSI-H and the discordance rate MMR-IHC and MSI-PCR in EMC decreased to 6.58% (15/228). No minimal microsatellite shift was found in CRC. Compared with EMC group with major microsatellite shift, cases with minimal microsatellite shift showed younger age, better tumor differentiation, and earlier International Federation of Gynecology and Obstetrics (FIGO) stage. There were significant differences in histological variant and FIGO stage between the two groups (P < 0.001, P=0.006). CONCLUSION: EMC was more prone to minimal microsatellite shift, which should not be ignored in the interpretation of MSI-PCR results. The combined detection of MMR-IHC and MSI-PCR is the most sensitive and specific method to capture MSI tumors.

[Blastic plasmacytoid dendritic cell neoplasm: A clinico-pathological retrospective analysis of thirteen cases].

OBJECTIVE: To investigate the clinicopathological features of blastic plasmacytoid dendritic cell neoplasm (BPDCN). METHODS: A total of 13 cases of BPDCN diagnosed in Peking University First Hospital from January 2013 to March 2022 were collected. The clinical features, histopathological characteristics, immunophenotypes and prognosis of the patients were analyzed retrospectively, and the related literatures was reviewed as well. RESULTS: Among the 13 patients, 11 were male and 2 were female, with a median age of 62 years (ranging from 5 to 78 years). Among them, single organ involvement occurred in 5 cases, all of which presented with skin lesions. Two or more organs were involved in other 8 cases (single organ with bone marrow involved in 3 cases; skin, bone marrow and lymph node involved simultaneously in 3 cases; skin, bone marrow, lymph node and spleen involved simultaneously in 2 cases). Histopathologically, it was characterized by the proliferation of medium to large atypical blastic cells, which infiltrated the whole thickness of dermis. When involved, the bone marrow lesions mainly appeared in a diffuse pattern, while the lymph node structure was usually destroyed, and the red pulp of the affected spleen was diffusely invaded. Immunohistochemical staining showed that all the 13 cases were positive for CD4, CD56, and CD123 (13/13) in varying degrees. All the 9 cases expressed TCL1 (9/9). Variable expression of CD68 (KP1) (8/13), TdT (7/12), CD117 (2/6), and high Ki-67 proliferation index (40%~80%) were showed. The neoplastic cells lacked expressions of CD20, CD3, MPO, CD34, or CD30; EBER in situ hybridization were negative (0/9). After definite diagnosis, 6 cases received chemotherapy, among which 1 received adjuvant radiotherapy, and 2 received subsequent bone marrow transplantation. Another 2 cases only received maintenance treatment. The median follow-up time was 14 months (ranging from 6 to 36 months), 5 patients died of the disease (6 to 18 months), 3 patients survived (7 to 36 months up to now), and the remaining 5 patients lost follow-up. CONCLUSION: BPDCN is a rare type of malignant lymphohematopoietic tumor with aggressive behavior and poor prognosis. The diagnosis should be made combining clinical features, histopathology, and immunohistochemical phenotype. Attention should be paid to differentiating BPDCN from other neoplasms with blastoid morphology or CD4+CD56+ tumors.

LINC00052 inhibits tumor growth, invasion and metastasis by repressing STAT3 in cervical carcinoma.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "LINC00052 inhibits tumor growth, invasion and metastasis by repressing STAT3 in cervical carcinoma, by J. Lin, L.-L. Nong, M.-Q. Li, F.-C. Yang, S.-H. Wang, M.-J. Liu, published in Eur Rev Med Pharmacol Sci 2019; 23 (11): 4673-4679-DOI: 10.26355/eurrev_201906_18047-PMID: 31210293" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/18047.

[Semiquantitative parameters of 18F-FDG PET/CT, gene mutation states of epidermal growth factor receptor and anaplastic lymphoma kinase in prognosis evaluation of patients with lung adenocarcinoma].

OBJECTIVE: To explore the valuable predictors for evaluating progression-free survival (PFS) in patients with lung adenocarcinoma, we analyzed the potential roles of standardized uptake value (SUV)-derived parameters from 18F-FDG PET/CT, combining with the gene mutation states of epidermal growth factor receptor (EGFR) and anaplastic lymphoma kinase (ALK), and other clinical characteristics. METHODS: Data of 84 lung adenocarcinoma patients pre-treated, who underwent 18F-FDG PET/CT scans, EGFR gene mutations test, ALK rearrangement assay and other relative tests, were retrospectively collected. Then a series of clinical parameters including EGFR/ALK mutation status and SUV-derived features [maximum standardized uptake value (SUVmax), average of standardized uptake value (SUVmean), metabolic tumor volume (MTV), and total lesion glycolysis (TLG)] were evaluated. Best possible cutoff points for all measuring parameters were calculated using receiver operating characteristic curve (ROC) analysis. Survival analysis was performed using Cox proportional hazards model to determine the prognostic markers for progression-free survival (PFS). Survival curves were obtained through Log-rank test and Kaplan-Meier curve. RESULTS: The median follow-up period was 31 months (24 to 58 months). It was found that SUVmax (≥3.01), SUVmean (≥2.25), MTV (≥25.41 cm3), and TLG (≥55.02) of the primary tumors were significantly associated with PFS in univariate Cox proportional hazards regression. Then regardless of age, gender, co-morbidity, EGFR/ALK mutation status, and treatment program, TLG (≥ 55.02, HR=4.965, 95%CI: 1.360-18.133), TNM stage (Ⅲ/Ⅳ, HR=7.811, 95%CI: 2.977-20.489), pro-gastrin releasing peptide (proGRP) (≥45.65 ng/L, HR=4.070, 95%CI: 1.442-11.487), tissue polypeptide antigen (TPA) (≥68.20 U/L, HR=6.996, 95%CI: 1.458-33.574), alkaline phosphatase (ALP) (≥82.50 IU/L, HR=4.160, 95%CI: 1.416-12.219) and ratio of activated partial thromboplastin time (aPTTR) (≥1.16: HR=4.58, 95%CI: 1.913-10.946) showed the independently relevant to PFS through multivariate Cox proportional hazards analysis. The EGFR mutant (P=0.343) and ALK rearrangement (P=0.608) were not significant either in survival analysis. CONCLUSION: High SUV-derived parameters (SUVmax, SUVmean, MTV and TLG) might provide prognostic value to some extent. Especially, TLG, and other clinical features [TNM stage, proGRP, TPA, ALP, and aPTTR] could be independently and significantly associated with PFS of lung adenocarcinoma patients. However, EGFR/ALK gene status could not be effectively relevant to PFS in lung adenocarcinoma patients.

[Clinicopathological features of lung salivary gland-type tumors].

Objective: To study the clinicopathological characteristics of lung salivary gland-type tumors (SGT), and to compare with the corresponding primary SGT in salivary glands. Methods: Twenty-three cases of lung SGT were retrieved from the files of Peking University First Hospital from January 2004 to September 2018. The morphology, immunophenotype, genotype and outcome of these cases were analyzed. Results: The 23 patients included 13 males and 10 females, with age range of 13-79 years (median 54 years). There were 11 cases of adenoid cystic carcinoma, 10 cases of mucoepidermoid carcinoma (MEC), one case each of clear cell carcinoma and myoepithelioma. The morphology and immunophenotype of lung SGT were very similar to their counterparts in salivary gland. MYB rearrangement was detected in one of 11 adenoid cystic carcinomas. MAML2 rearrangement was detected in all the MECs. EWSR1 rearrangement was detected in the one case of clear cell carcinoma. Of patients with adenoid cystic carcinoma, the survival time was more than 60 months (three cases), 52 months (one case), and 12-36 months (three cases). There was no recurrence and death in seven cases of MEC with follow-up results. One case of clear cell carcinoma recurred after 52 months of follow-up. Conclusions: Although the SGT of lung and their counterparts in salivary gland are very similar in their morphology, immunophenotype, genotype and prognosis, there are also some differences between each other. MYB rearrangement can be detected in most adenoid cystic carcinomas of salivary gland, but rarely in lung adenoid cystic carcinoma. The prognosis of patients with lung MEC is better than that of patients with salivary gland MEC, while the prognosis of patients with lung adenoid cystic carcinoma is worse than that of patients with salivary gland adenoid cystic carcinoma.

LINC00052 inhibits tumor growth, invasion and metastasis by repressing STAT3 in cervical carcinoma.

OBJECTIVE: The vital role of long noncoding RNAs (lncRNAs) in tumor progression has been identified in numerous studies. In this research, the biological function of lncRNA LINC00052 during the development of cervical cancer was mainly explored. PATIENTS AND METHODS: LINC00052 expression was detected by quantitative Real-time polymerase chain reaction (qRT-PCR) in cervical cancer tissue samples and cell lines. Moreover, the correlation between LINC00052 expression level and disease-free survival rate of cervical cancer patients was analyzed. In vitro functions of LINC00052 in cervical cancer cells were evaluated by proliferation assay, wound healing assay and transwell assay. In addition, qRT-PCR and Western blot were utilized to explore the underlying mechanism of LINC00052 in mediating the progression of cervical cancer. RESULTS: LINC00052 expression level was lower in cervical cancer samples than that in adjacent tissues, which was correlated with disease-free survival time. Moreover, cell proliferation, migration and invasion were inhibited through overexpression of LINC00052 in vitro. The mRNA and protein expression of signal transducers and activators of transcription 3 (STAT3) was downregulated after overexpressing LINC00052 in cervical cancer cells. The STAT3 expression level was negatively correlated with the expression of LINC00052 in cervical cancer tissues. CONCLUSIONS: LINC00052 could repress metastasis and invasion of cervical cancer cell via suppressing STAT3. LINC00052 might be a novel tumor suppressor in cervical cancer.

Effects of ulinastatin on inflammatory response and cognitive function after hip arthroplasty for the elderly patients with femoral neck fracture.

OBJECTIVE: To investigate the effects of ulinastatin on inflammatory response and cognitive function after hip arthroplasty for the elderly patients with femoral neck fracture. PATIENTS AND METHODS: A total of 80 patients with femoral neck fracture receiving hip arthroplasty in our hospital from August 2016 to February 2017 were selected and divided into observation group (n=40) and control group (n=40) using a random number table. The control group was treated with hip arthroplasty and symptomatic and supportive treatment after operation, while the observation group was treated with ulinastatin based on the treatment means of control group. The changes in antioxidant capacities, plasma noradrenaline (NA) and adrenaline (A) levels between the two groups before and after intervention were compared. The changes in neuron-specific enolase (NSE) and plasma S-100B protein levels before intervention and at 48 h after intervention were also compared. Moreover, the changes in mini-mental state examination (MMSE) scores during intervention and the Harris hip scores before intervention and at discharge between the two groups were compared. Finally, the off-bed walking time and postoperative discharge time of the two groups were recorded. RESULTS: After intervention, the levels of malondialdehyde (MDA) and superoxide dismutase (SOD) and the total antioxidant capacity in observation group were significantly superior to those in observation group before intervention and control group after intervention (p<0.05). After intervention, the levels of NA and A in observation group were lower than those in control group (p<0.05), and the levels of interleukin-1 (IL-1), tumor necrosis factor-α (TNF-α) and high-sensitivity C-reactive protein (hs-CRP) in observation group were also lower than those in control group (p<0.05). At 48 h after intervention, the levels of NSE and plasma S-100B protein in observation group were significantly lower than those in observation group before intervention and control group at 48 h after intervention (p<0.05). At 12 h, 24 h and 48 h after intervention, the MMSE scores of observation group were superior to those of control group in the same period (p<0.05). After intervention, the Harris hip score of observation group was superior to that of control group before and after intervention (p<0.05). The postoperative discharge time of observation group was earlier than that of control group (p<0.05), and the off-bed walking time was also earlier than that of control group (p<0.05). CONCLUSIONS: The combined application of ulinastatin could effectively reduce the oxidative stress and inflammatory response, improve the neurological functions, and promote the postoperative recovery in the elderly patients with femoral neck fracture after hip arthroplasty.

[Clinic pathological and ultrasonographic features of primary thyroid lymphoma].

Objective: To analyze the ultrasonographic and clinicopathological features of primary thyroid lymphoma(PTL). Methods: The ultrasonographic and clinicopathological featuresof 21 cases of pathologically-confirmed PTLs were analyzed retrospectively. Results: Of all 21 PTLs, 15 cases were diffuse large B-cell lymphoma, 4 were mucosal-associated lymphoid tissue extranodular marginal zone B-cell lymphoma, 1 was small B-cell lymphoma and 1 was classical Hodgkin lymphoma. Eight cases were proved by pathology with concomitant Hashimoto's thyroiditis. Ultrasonography observed bilateral or unilateral asymmetric goiter (21/21, 100.0%), marked hypoechogenicity (21/21, 100.0%) with posterior acoustic enhancement (19/21, 90.5%), heterogeneous echo texture with interspersed linear echogenic strands or intensive reticular echogenic strands or cloud echogenic, heterogeneous echo texture of thyroid gland (21/21, 100.0%), focal nodular hypoechoic(2/21, 9.5%) with regular or irregular shape, increased vascularity (13/21, 61.9%) and cervical lymphadenopathy (12/21, 57.1%). Two cases involved the anterior cervical muscle and 1 infiltrated trachea. Rapidly enlarging cervical mass were found in 13 cases (13/21, 61.9%)with associated compressive symptoms such as dyspnea, dysphagia and hoarseness. There was no any indisposed symptom in 3 cases. Conclusion: PTL has some common ultrasonographic and clinical features, core needle biopsy should be warranted to prove PTL.Surgical resection should be considered when needed to reduce misdiagnosis.

Comparison of lumbar pedicular dynamic stabilisation systems versus fusion for the treatment of lumbar degenerative disc disease: A meta-analysis.

This study aimed to systematically compare the safety, effectiveness and radiological changes after lumbar pedicular dynamic stabilisation systems and fusion to treat lumbar degenerative disc disease . All studies that were performed to compare various lumbar pedicular dynamic stabilisation systems with any lumbar fusion to treat lumbar degenerative disc disease and were published until April 30, 2015 were acquired through a comprehensive search in various databases. A meta-analysis was performed after the methodological qualities of trials were assessed and after data were extracted. Sixteen trials with 881 patients with a short-term follow-up (within 2 years) and a middle-term follow-up (2 to 4 years) were identified. Patients treated with lumbar pedicular dynamic stabilisation systems experienced more significant advantages in terms of operation time, intra-operative blood loss, complications and adjacent segment degeneration/disease development than those treated with lumbar fusion. The two groups did not significantly differ in terms of improvement in Oswestry Disability Index, visual analogue scale scores, satisfaction rate of operation and range of motion of adjacent segments. Lumbar pedicular dynamic stabilisation systems is superior to lumbar fusion to some extent, although some of its advantages have yet to be verified and compared with those of lumbar fusion. However, the two interventions were not significantly different in terms of relief in symptoms, functional recovery and motion preservation. Thus, lumbar pedicular dynamic stabilisation systems is recommended for its safety. A prudent attitude is necessary to choose between these interventions on the basis of effectiveness and changes in adjacent segments before a large-scale and long-term follow-up study can be performed.

Targeting PYK2 mediates microenvironment-specific cell death in multiple myeloma.

Multiple myeloma (MM) remains an incurable malignancy due, in part, to the influence of the bone marrow microenvironment on survival and drug response. Identification of microenvironment-specific survival signaling determinants is critical for the rational design of therapy and elimination of MM. Previously, we have shown that collaborative signaling between ß1 integrin-mediated adhesion to fibronectin and interleukin-6 confers a more malignant phenotype via amplification of signal transducer and activator of transcription 3 (STAT3) activation. Further characterization of the events modulated under these conditions with quantitative phosphotyrosine profiling identified 193 differentially phosphorylated peptides. Seventy-seven phosphorylations were upregulated upon adhesion, including PYK2/FAK2, Paxillin, CASL and p130CAS consistent with focal adhesion (FA) formation. We hypothesized that the collaborative signaling between ß1 integrin and gp130 (IL-6 beta receptor, IL-6 signal transducer) was mediated by FA formation and proline-rich tyrosine kinase 2 (PYK2) activity. Both pharmacological and molecular targeting of PYK2 attenuated the amplification of STAT3 phosphorylation under co-stimulatory conditions. Co-culture of MM cells with patient bone marrow stromal cells (BMSC) showed similar ß1 integrin-specific enhancement of PYK2 and STAT3 signaling. Molecular and pharmacological targeting of PYK2 specifically induced cell death and reduced clonogenic growth in BMSC-adherent myeloma cell lines, aldehyde dehydrogenase-positive MM cancer stem cells and patient specimens. Finally, PYK2 inhibition similarly attenuated MM progression in vivo. These data identify a novel PYK2-mediated survival pathway in MM cells and MM cancer stem cells within the context of microenvironmental cues, providing preclinical support for the use of the clinical stage FAK/PYK2 inhibitors for treatment of MM, especially in a minimal residual disease setting.

An updated meta-analysis of the Fc receptor-like 3 -169T/C polymorphism and rheumatoid arthritis risk.

OBJECTIVES: Published studies have shown conflicting results concerning the association between the -169T/C promoter polymorphism in the Fc receptor-like 3 (FCRL3) gene and rheumatoid arthritis (RA). In this study we conducted an up-to-date meta-analysis to examine the relationship. METHOD: We searched the PubMed database for all papers published up to 20 April 2012. Overall, 18 case-control studies with 12 620 cases and 12 613 controls were retrieved based on the search criteria for RA susceptibility related to the FCRL3 -169T/C polymorphism. Odds ratios (ORs) and 95% confidence intervals (CIs) were used to assess the strength of this association. Publication bias was assessed using the Egger test. RESULTS: We found that the FCRL3 -169T/C polymorphism increased the risk for RA overall in genetic models (allelic contrast: OR 1.09, 95% CI 1.03-1.14, p = 0.001; homozygote comparison: OR 1.20, 95% CI 1.08-1.34, p = 0.001; dominant genetic model: OR 1.03, 95% CI 1.01-1.05, p = 0.001). Stratified analysis by race also showed a significant positive association with Asians and Caucasians. Subgroup analysis of rheumatoid factor (RF) revealed a slightly positive relationship between the FCRL3 -169T/C polymorphism and RF-positive RA risk. No obvious evidence of publication bias was detected in the overall analysis. CONCLUSION: Our study indicates that the FCRL3 -169T/C polymorphism is significantly associated with increased RA risk.

CB1 and CB2 receptor mRNA expression in human peripheral blood mononuclear cells (PBMC) from various donor types.

Marijuana cannabinoid receptors (CBR), CB1 and CB2, are G protein-coupled receptors reported to be expressed in brain as well as cells of the periphery. Human peripheral blood mononuclear cells (PBMCs) are reported to express CBR mRNA with CB2 expression higher than CB1 and expression in B cells higher than other cells. However, it is not known if the mRNA expression is constant among individuals of differing ages, gender, or ethnic origins. In the present study, PBMCs were obtained from a limited number of normal donors of both genders, of ages ranging from 21 to 55, and from Caucasian, and Asian ethnic origin. Using semi-quantitative RT-PCR, we confirmed previous reports that CB2 mRNA expression was higher than CB1 in PBMCs and in addition demonstrated that this basic profile was observed when stratified by age, gender, or ethnic origin. The latter results suggest that CBR expression is relatively constant across the human population.

Expression of early lung cancer detection marker: hnRNP-A2/B1 and its relation to microsatellite alteration in non-small cell lung cancer.

We have reported that a mouse monoclonal antibody, 703D4, which recognizes heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNP-A2/B1) can frequently detect lung cancer in exfoliated sputum epithelial cells 1-2 years earlier than routine chest X-ray or sputum cytomorphology. We along with others have shown that microsatellite alteration (MA) at selected loci can be recognized in sputum cells prior to clinical lung cancer. The present study was undertaken to determine how frequently the expression of hnRNP-A2/B1 message is associated with neoplastic clonal expansion as shown by MA in 41 cases of non-small cell lung cancer (NSCLC). We used Northern blotting to evaluate hnRNP-A2/B1 mRNA expression in lung tumor and remote noninvolved lung. We evaluated microsatellite instability (i.e. shifts; MI) or loss of heterozygosity (LOH) with a panel of 13 microsatellite markers at loci identified previously as susceptible in NSCLC. Of the 41 tumors, 25 (61%) over-expressed hnRNP-A2/B1 and 33 (80%) demonstrated MA in at least one of 13 loci (58% in at least two loci). The association between MA (one locus) and the overexpression of hnRNP-A2/B1 is statistically significant (P=0.0082), and those lung tumors with MA at two or more loci were significantly more likely to over-express hnRNP-A2/B1 mRNA (P=0.004). MA of loci on 3p were the only MA statistically associated with hnRNP-A2/B1 message overexpression (P=0.001). We conclude that lung tumor cells undergoing clonal expansion frequently upregulate hnRNP-A2/B1.

[Studies on the efficacy of albendazole candy for treatment of intestinal nematode infections].

A single dose of albendazole (candy) 100 mg and 100 mg qd for two days was given respectively to 135 and 321 children who were infected with Enterobius vermicularis. All cases were cured 3wk later according to perianal tape examination. The same drug of 150 mg qd and 200 mg qd for two days was administered respectively to residents with single or multi-infections of Ascaris, hookworm and Trichuris; the egg negative conversion rates of the two dosages were revealed to be 99.4% (466/469) and 99.8% (487/488) for Ascaris infection, 96.8% (91/94) and 94.3% (99/105) for hookworm infection, and 53.4% (228/427) and 76.3% (370/486) for Trichuris infection. The maximal numbers of worm expelled were 100 on d2-3, 88 on d3-5 and 588 on d2-4 for Enterobius, Ascaris and hookworm, respectively; whereas the discharged Trichuris were scarce, merely 4.3 in average. The results exhibited promising efficacy of the drug on Enterobius, Ascaris and hookworm infections, but not so on trichuriasis.

Chloroplast DNA and cytoplasmic male-sterility.

This paper deals with the relationship of chloroplast DNA (ctDNA) and cytoplasmic malesterility (CMS) of maize, wheat and rape. Intramolecular heterogeneity and fragment patterns of DNA digested with restriction endonucleases were compared and analyzed. For this purpose methods of agarose gel electrophoresis and two dimensional gel electrophoresis with denaturating solvent concentration gradients were applied. 1) Heat denaturation of ctDNA from a male sterile line in maize yielded a profile with three melting regions. This base sequence heterogeneity was not observed for the ctDNA of its maintainer. 2) Maize ctDNA was digested with EcoR1 and BamH1 restriction endonucleases, wheat and rape ctDNA with EcoR1 only. No significant differences were observed in the comparison of the fragment patterns from the sterile lines and their maintainers with the exception of the sterile line in rape which lacked one fragment present in its maintainer. 3) Two dimensional electrophoresis using a gradient of denaturing agents revealed several differences in number and relative positions of the separated restriction endonuclease fragments between the sterile lines and their respective maintainers in all three tested crops. This raises the possibility that changes in ctDNA may be involved in CMS.