Objective To characterize the expression patterns of serum microRNA-497(miR-497)in patients with colorectal cancer(CRC)and to investigate its associations with clinicopathological characteristics,diagnostic performance,and long-term prognostic outcomes.Methods This study retrospectively analyzed data from 122 patients with CRC admitted to the hospital between March 2020 and March 2022(CRC group),and enrolled 100 healthy individuals undergoing routine physical examinations(healthy control group)for comparison.Serum samples were collected from all participants prior to any surgical intervention,and the expression levels of miR-497 in serum were quantified using real-time quantitative polymerase chain reaction(qRT-PCR).Simulta-neously,the levels of carcinoembryonic antigen(CEA)and carbohydrate antigen 199(CA199)were measured.To investigate the association between miR-497 expression and clinicopathological characteristics,we evaluated its diagnostic performance using receiver operating characteristic(ROC)curve analysis.Furthermore,Kaplan-Meier survival analysis and Cox proportional hazards regression models were employed to assess its impact on patient prognosis.Results Compared to healthy individuals,CRC patients exhibited significantly lower serum miR-497 expression levels(P<0.001).Notably,miR-497 expression was strongly correlated with TNM stage progression and lymph node metastasis(P<0.001),but showed no significant association with tumor location,patient sex,or age.Diagnostic evaluation using ROC curves demonstrated that miR-497 achieved an AUC of 0.845 for CRC detection,outperforming CEA(AUC=0.748)and CA19-9(AUC=0.702),with DeLong's test confirming the statistically significant differences(P<0.05).Kaplan-Meier survival analysis revealed a significantly higher 3-year DFS rate among patients with high miR-497 expression(84.06%)compared to those with low expression(64.58%),with median DFS not reached in the high-expression group versus 36 months in the low-expression group(P=0.015).Multivariate Cox regression analysis confirmed that reduced miR-497 expression(HR=1.923,95%CI:1.184～3.125),advanced TNM stage(HR=2.511,95%CI:1.421～4.437),and lymph node metastasis(HR=1.753,95%CI:1.151～2.664)were independently associated with poorer disease-free survival outcomes.Conclusions Serum miR-497 is downregulated in patients with CRC and is significantly associated with tumor progression and poor prognosis.It demonstrates high diagnostic accuracy and strong potential for prognostic evalua-tion,highlighting its promise as a biomarker for auxiliary diagnosis and outcome prediction in CRC.

Objective To characterize the expression patterns of serum microRNA-497(miR-497)in patients with colorectal cancer(CRC)and to investigate its associations with clinicopathological characteristics,diagnostic performance,and long-term prognostic outcomes.Methods This study retrospectively analyzed data from 122 patients with CRC admitted to the hospital between March 2020 and March 2022(CRC group),and enrolled 100 healthy individuals undergoing routine physical examinations(healthy control group)for comparison.Serum samples were collected from all participants prior to any surgical intervention,and the expression levels of miR-497 in serum were quantified using real-time quantitative polymerase chain reaction(qRT-PCR).Simulta-neously,the levels of carcinoembryonic antigen(CEA)and carbohydrate antigen 199(CA199)were measured.To investigate the association between miR-497 expression and clinicopathological characteristics,we evaluated its diagnostic performance using receiver operating characteristic(ROC)curve analysis.Furthermore,Kaplan-Meier survival analysis and Cox proportional hazards regression models were employed to assess its impact on patient prognosis.Results Compared to healthy individuals,CRC patients exhibited significantly lower serum miR-497 expression levels(P<0.001).Notably,miR-497 expression was strongly correlated with TNM stage progression and lymph node metastasis(P<0.001),but showed no significant association with tumor location,patient sex,or age.Diagnostic evaluation using ROC curves demonstrated that miR-497 achieved an AUC of 0.845 for CRC detection,outperforming CEA(AUC=0.748)and CA19-9(AUC=0.702),with DeLong's test confirming the statistically significant differences(P<0.05).Kaplan-Meier survival analysis revealed a significantly higher 3-year DFS rate among patients with high miR-497 expression(84.06%)compared to those with low expression(64.58%),with median DFS not reached in the high-expression group versus 36 months in the low-expression group(P=0.015).Multivariate Cox regression analysis confirmed that reduced miR-497 expression(HR=1.923,95%CI:1.184～3.125),advanced TNM stage(HR=2.511,95%CI:1.421～4.437),and lymph node metastasis(HR=1.753,95%CI:1.151～2.664)were independently associated with poorer disease-free survival outcomes.Conclusions Serum miR-497 is downregulated in patients with CRC and is significantly associated with tumor progression and poor prognosis.It demonstrates high diagnostic accuracy and strong potential for prognostic evalua-tion,highlighting its promise as a biomarker for auxiliary diagnosis and outcome prediction in CRC.

AIM To establish the HPLC fingerprints for Tanacetum tatsienense(Bureau & Franchet)K.Bremer & Humphries and to determine the contents of chlorogenic acid,quercetin-3-O-glucoside,luteolin-7-O-glucoside,luteolin-7-O-glucuronide,apigenin-7-O-glucuronide,luteolin and apigenin.METHODS The analysis was performed on a 35 ℃ thermostatic Agilent ZORBAX Extend C18 column(4.6 mm×250 mm,0.5 μm),with the mobile phase comprising of 0.5％phosphoric acid-acetonitrile flowing at 1.0 mL/min,and the detection wavelength was set at 350 nm.Subsequently,principal component analysis,partial least squares discriminant analysis and cluster analysis were carried out.RESULTS There were twelve common peaks in the fingerprints for fourteen batches of samples with the similarities of 0.761-0.975.Seven constituents showed good linear relationships within their own ranges(R2≥0.999 1),whose average recoveries were 84.00％-105.11％with the RSDs of 1.28％-2.86％.Various batches of samples were clustered into three types,and seven differential constituents were observable,containing peaks 4(luteolin-7-O-glucoside),12(apigenin),9(apigenin-7-O-glucuronide),8,11(luteolin),5(luteolin-7-O-glucuronide)and 2.CONCLUSION This precise,stable,specific and reproducible method can be used for the quality control of T.tatsienense.

AIM To study the chemical constituents from the methanol fraction of Premna fulva Craib leaves and their anti-inflammatory activities.METHODS The methanol fraction from P.fluva were isolated and purified by TLC,column chromatography,and HSCCC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.Their anti-inflammatory activities in vitro were evaluated by RAW264.7 model.RESULTS Twelve compounds were isolated and identified as vitexin(1),balanophonin(2),inotodisaccharide(3),4-allyl-2,6-dimethoxyphenol-β-D-glucopyranoside(4),dehydrovomifoliol(5),loliolide(6),(E)-4-((1S,3R,4R)-1-hydroxy-4,5,5-trimethyl-7-oxabicyclo[4.1.0]heptan-1-yl)but-1-en-3-o-ne(7),(E)-4-hydroxyphenylprop-7-ene 4-O-β-D-glucopyranoside(8),4-O-β-D-glucopyranosylbenzoic acid(9),vicenin(10),oleanicacid(11),sesamin(12),respectively.Compounds 1,2,5,7,10,and 12 showed good inhibitory activities against NO,and the IC50 values were(26.42±2.5)、(21.24±2.2)、(25.88±1.9)、(29.72±2.1)、(8.90±1.1)、(9.73±0.7)μmol/L,respectively.CONCLUSION Compounds 2,4-8 are isolated from Premna genus plants for the first time.Compounds 1,2,5,7,10,12 have anti-inflammatory activities.

Objective: Compare and analyze the results of the domestic Lanyi AH600 glycated hemoglobin analyzer and other different detection systems to understand the comparability of the detection results of different detectors, and establish the best cut point of Lanyi AH600 determination of haemoglobin A1c (HbA1c) in the diagnosis of diabetes. Methods: Multi center cohort study was adopted. The clinical laboratory departments of 18 medical institutions independently collected test samples from their respective hospitals from March to April 2022, and independently completed comparative analysis of the evaluated instrument (Lanyi AH600) and the reference instrument HbA1c. The reference instruments include four different brands of glycosylated hemoglobin meters, including Arkray, Bio-Rad, DOSOH, and Huizhong. Scatter plot was used to calculate the correlation between the results of different detection systems, and the regression equation was calculated. The consistency analysis between the results of different detection systems was evaluated by Bland Altman method. Consistency judgment principles: (1) When the 95% limits of agreement (95% LoA) of the measurement difference was within 0.4% HbA1c and the measurement score was≥80 points, the comparison consistency was good; (2) When the measurement difference of 95% LoA exceeded 0.4% HbA1c, and the measurement score was≥80 points, the comparison consistency was relatively good; (3) The measurement score was less than 80 points, the comparison consistency was poor. The difference between the results of different detection systems was tested by paired sample T test or Wilcoxon paired sign rank sum test; The best cut-off point of diabetes was analyzed by receiver operating characteristic curve (ROC). Results: The correlation coefficient R² of results between Lanyi AH600 and the reference instrument in 16 hospitals is≥0.99; The Bland Altman consistency analysis showed that the difference of 95% LoA in Nanjing Maternity and Child Health Care Hospital in Jiangsu Province (reference instrument: Arkray HA8180) was -0.486%-0.325%, and the measurement score was 94.6 points (473/500); The difference of 95% LoA in the Tibetan Traditional Medical Hospital of TAR (reference instrument: Bio-Rad Variant II) was -0.727%-0.612%, and the measurement score was 89.8 points; The difference of 95% LoA in the People's Hospital of Chongqing Liang Jiang New Area (reference instrument: Huizhong MQ-2000PT) was -0.231%-0.461%, and the measurement score was 96.6 points; The difference of 95% LoA in the Taihe Hospital of traditional Chinese Medicine in Anhui Province (reference instrument: Huizhong MQ-2000PT) was -0.469%-0.479%, and the measurement score was 91.9 points. The other 14 hospitals, Lanyi AH600, were compared with 4 reference instrument brands, the difference of 95% LoA was less than 0.4% HbA1c, and the scores were all greater than 95 points. The results of paired sample T test or Wilcoxon paired sign rank sum test showed that there was no statistically significant difference between Lanyi AH600 and the reference instrument Arkray HA8180 (Z=1.665,P=0.096), with no statistical difference. The mean difference between the measured values of the two instruments was 0.004%. The comparison data of Lanyi AH600 and the reference instrument of all other institutions had significant differences (all P<0.001), however, it was necessary to consider whether it was within the clinical acceptable range in combination with the results of the Bland-Altman consistency analysis. The ROC curve of HbA1c detected by Lanyi AH600 in 985 patients with diabetes and 3 423 patients with non-diabetes was analyzed, the area under curve (AUC) was 0.877, the standard error was 0.007, and the 95% confidence interval 95%CI was (0.864, 0.891), which was statistically significant (P<0.001). The maximum value of Youden index was 0.634, and the corresponding HbA1c cut point was 6.235%. The sensitivity and specificity of diabetes diagnosis were 76.2% and 87.2%, respectively. Conclusion: Among the hospitals and instruments currently included in this study, among these four hospitals included Nanjing Maternity and Child Health Care Hospital in Jiangsu Province (reference instrument: Arkray HA8180), Tibetan Traditional Medical Hospital of TAR (reference instrument: Bio-Rad Variant Ⅱ), the People's Hospital of Chongqing Liang Jiang New Area (reference instrument: Huizhong MQ-2000PT), and the Taihe Hospital of traditional Chinese Medicine in Anhui Province (reference instrument: Huizhong MQ-2000PT), the comparison between Lanyi AH600 and the reference instruments showed relatively good consistency, while the other 14 hospitals involved four different brands of reference instruments: Arkray, Bio-Rad, DOSOH, and Huizhong, Lanyi AH600 had good consistency with its comparison. The best cut point of the domestic Lanyi AH600 for detecting HbA1c in the diagnosis of diabetes is 6.235%.
Pregnancy ; Child ; Humans ; Female ; Glycated Hemoglobin ; Cohort Studies ; Diabetes Mellitus/diagnosis* ; Sensitivity and Specificity ; ROC Curve

Objective: To analyze the genotypes and clinical phenotypes of patients with epilepsy associated with IQSEC2 gene variants. Methods: The genotypes, seizure types, electroencephalogram, neuroimage of 6 patients with IQSEC2 gene variants in the Department of Pediatrics, Peking University First Hospital from July 2019 to October 2021 were analyzed. Results: There were 5 males and 1 female. Six variants were de novo, including 2 frameshift variants (c.3801_3808dup/p.Q1270Rfs*130, c.1459_1460delAT/p.M487Vfs*2), 2 nonsense variants (c.3163C>T/p.R1055*, c.1417G>T/p.E473*), 1 in-frame deletion (c.2295_2297del/p.N765del) and 1 missense variant (c.2293A>G/p.N765D). Age at seizure onset ranged from 3 months to 2 years and 5 months. Multiple seizure types were observed, including epileptic spasms in 5 patients, focal seizures in 5 patients, tonic seizures in 3 patients, myoclonic seizures in 3 patients, atypical absence seizures in 2 patients and atonic seizures in 2 patients. All 6 patients showed global developmental delay before seizure onset. There were other clinical manifestations, including autistic features in 3 patients, microcephaly in 3 patients, dystonia in 2 patients and binocular esotropia in 1 patient. The electroencephalogram showed slow background activity and hypsarrhythmia in all 6 patients. Brain magnetic resonance imaging showed abnormal in 5 patients and normal in 1 patient. Five patients were diagnosed with infantile spasms. Among them, 4 patients had late-onset infantile spasms. One patient was unclassified developmental epileptic encephalopathy. The age of last follow-up ranged from 3 years and 2 months to 7 years and 2 months. All 6 patients still had seizures after multiple anti-seizure medications. Conclusions: The seizure onset of patients with IQSEC2 gene variants usually begins after 1 year of age. The common seizure types include epileptic spasms and focal seizures. Patients usually have a global developmental delay before seizure onset. IQSEC2 variants could be related to developmental and epileptic encephalopathy, and most patients are diagnosed with late onset infantile spasms. Epilepsy associated with IQSEC2 gene variants is usually refractory.
Female ; Male ; Child ; Humans ; Spasms, Infantile/genetics* ; Genotype ; Phenotype ; Epilepsy/genetics* ; Seizures ; Spasm ; Guanine Nucleotide Exchange Factors

Humans ; Carcinoma, Hepatocellular/secondary* ; Liver Neoplasms/surgery* ; Thyroid Neoplasms/surgery* ; Neoplasms, Second Primary

Objective: To investigate the incidence and treatment of perioperative anemia in patients with gastrointestinal neoplasms in Hubei Province. Methods: The clinicopathological data of 7 474 patients with gastrointestinal neoplasms in 62 hospitals in 15 cities (state) of Hubei Province in 2019 were collected in the form of network database. There were 4 749 males and 2 725 females. The median age of the patients was 62 years (range: 17 to 96 years). The hemoglobin value of the first time in hospital and the first day after operation was used as the criterion of preoperative anemia and postoperative anemia. Anemia was defined as male hemoglobin <120 g/L and female hemoglobin <110.0 g/L, mild anemia as 90 to normal, moderate anemia as 60 to <90 g/L, severe anemia as <60 g/L. The t test and χ² test were used for inter-group comparison. Results: The overall incidence of preoperative anemia was 38.60%(2 885/7 474), and the incidences of mild anemia, moderate anemia and severe anemia were 25.09%(1 875/7 474), 11.37%(850/7 474) and 2.14%(160/7 474), respectively. The overall incidence of postoperative anemia was 61.40%(4 589/7 474). The incidence of mild anemia, moderate anemia and severe anemia were 48.73%(3 642/7 474), 12.20%(912/7 474) and 0.47%(35/7 474), respectively. The proportion of preoperative anemia patients receiving treatment was 26.86% (775/2 885), and the proportion of postoperative anemia patients receiving treatment was 14.93% (685/4 589). The proportions of preoperative anemia patients in grade ⅢA, grade ⅢB, and grade ⅡA hospitals receiving treatment were 26.12% (649/2 485), 32.32% (85/263), and 29.93% (41/137), and the proportions of postoperative anemia patients receiving treatment were 14.61% (592/4 052), 22.05% (73/331), and 9.71% (20/206). The proportion of intraoperative blood transfusion (16.74% (483/2 885) vs. 3.05% (140/4 589), χ²=434.555, P<0.01) and the incidence of postoperative complications (17.78% (513/2 885) vs. 14.08% (646/4 589), χ²=18.553, P<0.01) in the preoperative anemia group were higher than those in the non-anemia group, and the postoperative hospital stay in the preoperative anemia group was longer than that in the non-anemia group ((14.1±7.3) days vs. (13.3±6.2) days, t=5.202, P<0.01). Conclusions: The incidence of perioperative anemia in patients with gastrointestinal neoplasms is high. Preoperative anemia can increase the demand for intraoperative blood transfusion and affect the short-term prognosis of patients. At present, the concept of standardized treatment of perioperative anemia among gastrointestinal surgeons in Hubei Province needs to be improved.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anemia/epidemiology* ; Blood Transfusion ; Female ; Gastrointestinal Neoplasms/surgery* ; Humans ; Length of Stay ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVES: To construct a cell line that can stably express human phospholamban(PLN) and initially explore its application in the study of myocardial toxicity mechanism. METHODS: FastCloning method was used to insert the open reading frame sequence of target gene PLN into eukaryotic expression vector pcDNA5/FRT/TO(hereinafter referred to as pDFT) to construct the pDFT-PLN-Flag plasmid. The Flp-In^TM T-REx^TM 293 cells were generated by cotransfection of the constructed plasmid and pOG44 plasmid to express the target gene. Successfully recombined monoclonal cell lines were screened by hygromycin B resistance. Western blot and indirect immunofluorescence (IFA) were used to examine the expression of the target protein in recombinant cells. After the cell line was exposed to aconitine, it was verified by Western blot to detect changes in PLN protein phosphorylation. RESULTS: After PCR amplification of the recombinant plasmid and DNA electrophoresis, the length of the amplified product is the same as the known PLN gene fragment, which is consistent with the open reading frame (ORF) sequence of the human PLN gene after sequencing. IFA and Western blot showed that the constructed proliferation cell line can stably express high levels of human PLN under induction and regulation. Preliminary results showed that the phosphorylation level of Thr17-PLN decreased after two hours of exposure to 1 μmol/L aconitine. CONCLUSIONS This human cell line can stably express PLN and can be used to study the mechanism of action of aconitine on the cell at molecular level.
Calcium-Binding Proteins/metabolism* ; Cell Line ; Humans ; Myocardium/metabolism* ; Phosphorylation

β-amyloid (Aβ) plaque deposition, neurofibrillary tangles and neuroinflammation are important pathological features of Alzheimer's disease (AD). Microglial membrane receptors, such as triggering receptor expressed on myeloid cells 2, Toll-like receptors, complement system and scavenger receptor, play a key role that mediate neuroinflammatory responses, promote Aβ clearance and signal transduction.