Objective: To investigate the effects of different storage temperatures and durations on the activities of coagulation factor Ⅷ (Factor Ⅷ, FⅧ) and coagulation factor Ⅸ (Factor Ⅸ, FⅨ) after whole blood collection, so as to provide data support for the optimal storage conditions. Methods: A total of 16 mL of whole blood was collected from each of the 20 healthy volunteers at our blood center and aliquoted into 8 sodium citrate anticoagulant tubes. Two tubes were immediately centrifuged for the measurement of FⅧ and FⅨ activity levels. The remaining 6 tubes of whole blood were respectively stored under room temperature and low-temperature conditions. At 2, 4, and 6 h, the whole blood samples were centrifuged and analyzed for FⅧ and FⅨ activity levels. The mean values of the two immediately tested tubes were used as the control group, while other tubes were designated as the experimental groups for comparison. Statistical analysis was performed using SPSS 26.0. Results: The activity of FⅧ in whole blood remained stable after 4 hours of storage at both room temperature and low temperature (116.53±25.95 vs 125.22±27.33, 109.77±23.23 vs 125.22±27.33) (P>0.05 for both). However, by 6 hours, FⅧ activity showed a statistically significant decline compared to the control group (108.65±22.92 vs 125.22±27.33, 100.46±20.19 vs 125.22±27.33) (P<0.05 for both), though the room temperature group results were closer to the control values. The activity of FⅨ in whole blood remained stable after 6 hours of storage under both conditions (97.14±19.48 vs 96.76±19.67, 97.10±17.45 vs 96.76±19.6) (P>0.05 for all comparisons). Conclusion: For whole blood samples after collection, storage at either room temperature or low temperature for up to 4 hours does not compromise the accuracy of test results. When stored for 6 hours, FⅨ activity remains stable, whereas FⅧ activity decreases significantly. Notably, FⅧ activity demonstrates better stability at room temperature than under low-temperature conditions within the 6-hour storage.

Objective: To study the correlation between activated partial thromboplastin time (APTT) mixing test results and the inhibitor titers in hemophilia A inhibitor-positive patients. Methods: In this cross-sectional study, 41 patients with severe hemophilia A and inhibitors (and negative for lupus anticoagulant) were included from the hemophilia clinic of Shandong Blood Center from February 2022 to February 2024. All patients underwent APTT mixing test. The Rosner's index (RI, including the immediate RI and the RI after 2-hour water bath incubation [water bath 2h RI]), the time-dependent difference (Δ value), and the corrected percentage were calculated based on results of APTT mixing test. The median (interquartile range) of the corresponding indexes were calculated, and the ROC curves for identification of high inhibitor titers using the four indexes (the immediate RI, the water bath 2h RI, the Δ value, and the corrected percentage) were plotted, The correlations between APTT mixing test and inhibitor titers for coagulation factor Ⅷ (Factor Ⅷ, FⅧ) were investigated. Results: The median (lower quartile, upper quartile) of immediate RI, water bath 2h RI, Δ-value and corrected percentage for FⅧ inhibitor positive patients were 11.0 (5.4, 29.3)%, 45.0 (25.7, 75.0)%, 26.2 (7.6, 41.8) s, and 82.2 (58.5, 91.6)%, respectively. The median (lower quartile, upper quartile) of the immediate RI, water bath 2h RI, Δ-value and corrected percentage were 25.2 (13.0, 37.5)%, 64.1 (44.6, 72.6)%, 38.0 (14.3, 38.3) s, and 66.5 (50.1, 82.1)% for the high-titer inhibitor group, and 5.2 (4.2, 9.4)%, 17.9 (8.8, 28.0)%, 13.0 (7.6, 25.4) s, and 92.3 (88.0, 94.3)% for the low-titer inhibitor group. The AUCs of the ROC curves for discrimination between high and low titer inhibitor were: 0.9105 for immediate RI, 0.9118 for water bath 2h RI, 0.8873 for correcter percentage, and 0.6532 for Δ-value. Conclusion: High-titer inhibitors can be highly suspected in hemophiliac patients with an immediate RI >10% and a water bath 2h RI >45%, and the presence of low-titer inhibitors is suspected in patients with a 4-second < immediate RI <10% and a 13% < water bath 2h RI <45%.

Objective:To analyze the clinicopathological features of prostate cancers with BRCA2 pathogenic mutations, and the association between BRCA2 pathogenic mutation and clinicopathological characteristics. Patient survivals were also examined.Methods:Clinicopathological data of 249 prostate cancer patients who underwent genetic testing in West China Hospital of Sichuan University, Chengdu, China from June 2014 to August 2021 were collected. A retrospective analysis of histopathological morphology, clinicopathological characteristics, and patient survivals was conducted.Results:The genetic testing in the 249 prostate cancer patients showed a pathogenic mutation of DNA damage repair gene (DRG) in 73 cases (73/249, 29.3%), including 22 cases (8.8%) with BRCA2 pathogenic mutation and 51 cases with pathogenic mutations of other DRG. Among the 22 patients with BRCA2 pathogenic mutation, 14 patients (5.6%) harbored germline mutations and 8 patients (3.2%) somatic mutations. Their ages ranged from 48 to 91 years, with a median of 67 years. Seventeen patients (77.3%) had distant metastasis, including 16 cases with bone metastasis and 1 case with multiple metastases. Thirteen patients (59.1%) were castration-resistant prostate cancer. The histological type was mainly classical prostatic acinar adenocarcinoma, including 16 cases (72.7%) with intraductal carcinoma of the prostate (IDC-P). Six cases (27.3%) showed focal neuroendocrine differentiation. Perineural/vascular invasion and extraprostatic extension were seen in 11 cases (50.0%) and 8 cases (36.4%), respectively. The Gleason scores of 19 patients (86.4%) were≥8. IDC-P was more commonly found in patients with BRCA2 germline pathogenic mutation than those with BRCA2 somatic pathogenic mutation, other DRG pathogenic mutation or no-DRG pathogenic mutation ( P=0.002). With a total follow-up time of 189 months, the median overall survival (OS) was 132.3 months. Patients with DRG pathogenic mutation had shorter OS than those with no-DRG pathogenic mutation ( P=0.040). The OS of patients with BRCA2 germline pathogenic mutation did not significantly differ from that of patients with BRCA2 somatic pathogenic mutation, other DRG pathogenic mutation or no-DRG pathogenic mutation ( P=0.216). Conclusions:The presence of BRCA2 gene pathogenic mutation is common in the prostate cancers with high Gleason grade, advanced clinical stage, and castration resistance. IDC-P is more commonly noted in cases with BRCA2 germline pathogenic mutation than those without. Patients with DRG pathogenic mutation have shorter OS than those with no-DRG pathogenic mutation, but there is no significant association between BRCA2 pathogenic mutations and OS.

Na⁺/H⁺ antiporter (NHX) gene subfamily plays an important role in plant response to salt stress. In this study, we identified the NHX gene family members of Chinese cabbage and analyzed the expression patterns of BrNHXs gene in response to abiotic stresses such as high temperature, low temperature, drought and salt stress. The results showed that there were 9 members of the NHX gene family in Chinese cabbage, which were distributed on 6 chromosomes respectively. The number of amino acids was 513-1 154 aa, the relative molecular weight was 56 804.22-127 856.66 kDa, the isoelectric point was 5.35-7.68. Members of BrNHX gene family mainly existed in vacuoles, the gene structure is complete, and the number of exons is 11-22. The secondary structures of the proteins encoded by the NHX gene family in Chinese cabbage had alpha helix, beta turn and random coil, and the alpha helix occurred more frequently. Quantitative real-time PCR (qRT-PCR) analysis showed that the gene family members had different responses to high temperature, low temperature, drought and salt stress, and their expression levels differed significantly in different time periods. BrNHX02 and BrNHX09 had the most significant responses to these four stresses, and their expression levels were significantly up-regulated at 72 h after treatments, which could be used as candidate genes to further verify their functions.
Genome, Plant ; Multigene Family ; Stress, Physiological/genetics* ; Brassica/metabolism* ; Gene Expression Regulation, Plant ; Phylogeny ; Plant Proteins/metabolism*

Objective:To study the clinicopathological features, immunophenotype, molecular genetic characteristics and prognosis of renal cell carcinoma associated with TFEB gene rearrangement (TFEBr-RCC).Methods:Eight cases of TFEBr-RCC diagnosed at the West China Hospital of Sichuan University from 2014 to 2022 were collected for clinicopathological, immunohistochemical, fluorescence in situ hybridization and RNA sequencing analyses, with review of literature.Results:Six patients were male and two were female. The patient ages ranged from 25 to 50 years (mean: 34 years, median: 32 years). The tumors were present in the right kidney (3 cases) or the left kidney (5 cases). The maximum diameters of the tumors ranged from 4.0 cm to 18.5 cm, with an average of 8.5 cm. Histologically, majority of the cases (5/8) showed typical biphasic "pseudorosette" structure, while the remaining three cases demonstrated atypical morphology that was similar to epithelioid angiomyolipoma or clear cell renal cell carcinoma. Immunohistochemical study showed positivity of TFEB (8/8), PAX8 (8/8), MART-1 (7/7), and HMB45 (5/6). Interestingly, PD-L1 was variably expressed in all five tested cases. Staining for TFE3 in all cases was negative. TFEB translocation was verified in all 8 cases using TFEB fluorescence in situ hybridization. RNA sequencing showed MALAT1-TFEB gene fusion in 4 of the 5 tested cases (two of which showing novel MALAT1-TFEB fusion sites), and one case with a novel ACTB-TFEB gene fusion. Patient follow-ups ranged from 5 to 96 months (average 47 months). All patients were alive without recurrence or metastasis.Conclusions:TFEBr-RCC tends to occur in young adults and has a good prognosis. Histologically, most of the cases show characteristic biphasic structure, and some cases show epithelioid angiomyolipoma-like or clear cell RCC-like morphology. Immunohistochemical reactivity to TFEB, melanocytic markers and PD-L1 is characteristic. MALAT1-TFEB gene fusion is the most common molecular change, with variable fusion sites.

Objective:To investigate the expression of HER2 and its relationship with clinicopathological features in patients with urothelial carcinoma.Methods:Urothelial carcinoma specimens collected from January 2019 to June 2022 were used. The expression of HER2, cytokeratin 20 and cytokeratin 5/6 was examined using immunohistochemistry. The HER2 expression was assessed according to the clinical pathological expert consensus on HER2 testing in urothelial carcinoma in China. Cases with HER2 2+/3+ were classified as HER2 positive. The relationship between HER2 expression and clinicopathological and molecular features was analyzed.Results:Four hundred and twenty-nine urothelial carcinoma specimens were analyzed, including 166 cases of raclical resection and 263 cases of local tumor resection. The median patient-age was 69 years (range: 31-93 years). The male: female ratio was 2.9∶1.0. The positive rate of HER2 was 45.7%(196/429). The positive rate of HER2 in patients with local tumor resection was higher than that in patients with radical resection [51.7%(136/263) vs.36.1%(60/166), P<0.05]. In the upper urinary tract (renal pelvis/ureter) urothelial carcinomas, the positive rate of HER2 was 35.2% (37/105). In bladder urothelial carcinoma, the positive rate of HER2 was 49.1%(157/320), and higher than that in upper urinary tract urothelial carcinoma ( P<0.05). In high grade urothelial carcinoma, the positive rate of HER2 was 52.8%(168/318) and higher than that in low grade urothelial carcinomas (25.2%, 28/111, P<0.01). In 166 radical resection specimens, the positive rate of HER2 was not differentially distributed by tumor pT stage [Ta (26.1%, 6/23), T1 (41.7%, 20/48), T2 (40.0%, 10/25), T3 (28.1%, 16/57), T4 (8/13) ( P>0.05)]. In urothelial carcinomas with muscle invasion, the HER2 positive rate was 35.8%(34/95), while the rate in non-muscle-invasion urothelial carcinoma was 36.6%(26/71, P>0.05). CK20 and CK5/6 were used to refine the urothelial carcinoma molecular subtypes. The positive rate of HER2 was highest in CK20 +/CK5/6 -group (124/194, 63.9%), followed by CK20 +/CK5/6 +group (18/40, 45.0%), CK20 -/CK5/6 -group (14/41, 34.1%) and CK20 -/CK5/6 +group (25/131, 19.1%, P<0.01). The positive rate of HER2 in micropapillary urothelial carcinoma was highest (14/15), followed by urothelial carcinoma with glandular differentiation (11/14), conventional urothelial carcinoma (161/360, 44.7%) and urothelial carcinoma with squamous differentiation (6/35, 17.1%, P<0.01). In the cases with lymph node metastasis, the positive rate of HER2 was 45.5% (10/22) and higher than the cases without lymph node metastasis (31.0%, 13/42). But there was no statistically significant association between HER2 expression and lymph node metastasis ( P>0.05). Conclusions:Expression of HER2 in urothelial carcinoma is closely correlated with tumor location, grade, histologic subtypes, molecular subtypes and surgical approach, but not with pT stage, muscle invasiveness or lymph node metastasis.

Objective:To study the key factors that affect patients′ perception among the five dimensional factors of patients′ perceived service quality, and explore how to effectively rebuild patients′ confidence in the process of handling services with quality defects.Methods:A total of 388 12345 work orders from a Beijing stomatological hospital in 2021 were collected. The problems and solutions of patients′feedback were classified and standardized into tangibility, reliability, responsiveness, assurance and empathy. Chi-square test and logistic regression were used to analyze the relationship between relevant factors and problem solving rate, patient satisfaction.Results:There were 513 feedback questions in 388 return visit work orders, with 1.32 items for each. There were 83, 112, 126, 111 and 81 questions in the five dimensions of tangibility, reliability, responsiveness, assurance and empathy; 273 work orders had single-dimension problems, accounting for 70.4%, 105 work orders had two dimensional problems, and 10 work orders had three dimensional problems. Among them, the assurance dimension problem accounts for 47% and 100% of the work orders of two-dimension and three-dimension problems respectively; The proportion of unsolved responsiveness dimension problems was the highest, accounting for 31.7%, and there was a negative correlation between responsiveness dimension problems and patient satisfaction rate( r=-0.709). Conclusions:In the process of medical service, the basic quality and skills of medical staff are the basis of building patient trust. In the process of handling medical services with quality defects, it is most important that quickly respond to patients′ queries, which could help to rebuild patients′ confidence in the quality of services.

Objective:To investigate the clinical and pathological features and prognosis of patients with microfocal prostate adenocarcinoma.Methods:Clinical and pathological data of the patients diagnosed with microfocal adenocarcinoma on prostate biopsy at the West China Hospital from 2013 to 2019 were collected. Microfocal adenocarcinoma was defined as follows: Gleason score of 3+3=6, total number of the cores ≥10, number of the positive cores ≤2, and proportion of the tumor in each positive core<50%. Clinicopathological parameters, treatment plans and follow-up data were collected. Pathological information of the biopsy and radical resection specimens was used to analyze the correlation between pathological parameters in the biopsy report and adverse pathological features of radical resection specimens, including increased Gleason score, capsule invasion, positive surgical margin and perineural invasion.Results:A total of 206 cases of microfocal adenocarcinoma were diagnosed on prostate biopsies from 2013 to 2019, accounting for 6.7% of all adenocarcinoma cases. There were 139 cases of 1 positive core and 67 cases of 2 positive cores. Patients with microfocal adenocarcinoma were younger than those with non-microfocal adenocarcinoma (69 years versus 71 years, P<0.001). Compared with patients with non-microfocal adenocarcinoma, the pre-biopsy total prostate specific antigen (tPSA) and free prostate specific antigen (fPSA) levels in patients with microfocal adenocarcinoma were both lower (11.2 μg/L 2 versus 23.7 μg/L 2; 1.4 μg/L 2 versus 3.0 μg/L 2, P<0.001), the fPSA/tPSA level was higher (12.9% versus 10.7%, P<0.05), the prostate volume was larger (38.9 mL versus 34.3 mL, P<0.05), and the PSA density was lower (0.3 μg/L 2 versus 0.8 μg/L 2, P<0.001). 130 patients underwent radical prostatectomy, 30 patients chose active monitoring, 31 patients chose endocrine or radiation therapy, and 15 patients were lost to follow-up. Three patients in the active surveillance group underwent radical prostatectomy for disease progression after 21-39 months observation. Biochemical relapses occurred in two patients in the radical prostatectomy group. The remaining patients have no disease progression or recurrence at present. Compared with radical prostatectomy specimens, Gleason score in the biopsy material was increased in 64/115 patients (55.7%). Among resection excision specimens, 14 cases (12.2%) had extraprostatic extension (EPE), 35 cases (30.4%) had perineural invasion, and 16 cases (13.9%) had a positive margin. Univariate and multivariate analyses showed that low fPSA/tPSA ratio and 2 positive cores were independent risk factors for Gleason score increase in the radical prostatectomy specimens. A low fPSA/tPSA ratio was an independent risk factor for perineural invasion. Low fPSA/tPSA ratio and low prostate volume were associated with a positive margin in radical prostatectomy specimens. Conclusions:In this study, patients diagnosed with microfocal adenocarcinoma on prostate biopsy account for a high proportion of the patients with increased Gleason score in the radical prostatectomy specimens, and there is a certain proportion of adverse pathological features in the radical specimens. Therefore, for the patients with only a small amount of low-grade adenocarcinoma found in biopsy, PSA levels and PSA density should be taken into consideration in treatment selection.

Objective:To establish a continuous monitoring system based on nursing sensitivity outcome indexes so as to improve the quality of fall prevention in elderly inpatients.Methods:By adopting the convenient sampling method, the elderly inpatients and their caregivers in 14 wards of the Institute of Geriatrics of Guangdong People's Hospital from April 2016 to January 2017 were selected as the research objects, and the nurses in 14 wards were also included in the research. A total of 14 wards were randomly divided into the control group with 7 wards and the experimental group with 7 wards. In the experimental group, there were 151 patients, 137 caregivers and 68 nurses. In the control group, there were 151 patients, 131 caregivers and 57 nurses. In the control group, the quality of fall prevention was monitored by the conventional three-level monitoring method of quality control department of the nursing department, head nurse of department and head nurse of ward. On this basis, the experimental group applied Fall-related Evaluation Scale for Elderly Inpatients to continuously monitor the quality of fall prevention. Modified Fall Efficacy Scale (MFES) was used to compare the fall performance scores and the incidence of falls during hospitalization between the two groups. Caregivers' Fall Knowledge, Attitude and Practice Scale was used to compare the knowledge, belief, attitude and practice of caregivers in the two groups before and after intervention. The General Self-Efficacy Scale was used to compare the self-efficacy of nurses in the two groups. The Fall-Related Evaluation Scale for Elderly Inpatients was used to compare the quality evaluation results of fall prevention before and after intervention in the experimental group.Results:None of the patients in the experimental group fell during their hospitalization, and 2 patients in the control group fell. There was no statistically significant difference in the incidence of falls between the two groups ( P>0.05) . The fall efficacy score at discharge was (70.77±16.85) in the experimental group and (62.77±18.78) in the control group, and the difference was statistically significant ( t=3.179, P<0.01) . After intervention, the scores of fall knowledge, attitude and practice of caregivers in the experimental group were higher than those in the control group, and the self-efficacy score of nurses was higher than that in the control group, and the differences were statistically significant ( P<0.01) . The quality score of fall prevention in the experimental group after intervention was higher than that before intervention, and the difference was statistically significant ( P<0.05) . Conclusions:The establishment of continuous monitoring system based on the nursing sensitivity outcome index can improve the quality and efficacy of fall prevention in elderly patients and improve the knowledge, attitude and practice of the caregiver and the self-efficacy of the nurses.

Objectives:To summarize the clinical and genetic characteristics of the patients with isolated methylmalonic acidemia and investigate the strategies for the diagnosis, treatment and prevention.Methods:Three hundred and fourteen patients (180 males, 134 females) with isolated methylmalonic acidemia were ascertained from 26 provinces or cities across the mainland of China during January 1998 to March 2020. Genetic analysis was performed by Sanger sequencing, gene panel sequencing, whole exome sequencing, multiplex ligation-dependent probe amplification or quantitative PCR. According to the age of onset, the patients were divided to early-onset group (≤12 months of age) and the late-onset group (>12 months of age). They were treated by cobalamin, L-carnitine and (or) special diet and symptomatic treatment. Statistical analysis was done using Chi-square test.Results:Fifty-eight of 314 (18.5%) patients were detected by Newborn screening using liquid chromatography tandem mass spectrometry. Five cases (1.6%) had a postmortem diagnosis. Two hundred and fifty-one patients (79.9%) were clinically diagnosed with an age of onset ranged from 3 hours after birth to 18 years. One hundred and fifty-nine patients (71.0%) belonged to early-onset groups, 65 patients (29.0%) belonged to the late-onset group. The most common symptoms were metabolic crises, psychomotor retardation, epilepsy, anemia and multiple organ damage. Metabolic acidosis and anemia were more common in early-onset patients than that in late-onset patients (20.8%(33/159) vs. 9.2% (6/65), 34.6% (55/159) vs. 16.9% (11/165), χ 2=4.261, 6.930, P=0.039, 0.008). Genetic tests were performed for 236 patients (75.2%), 96.2%(227/236) had molecular confirmation. One hundred and twenty-seven variants were identified in seven genes (MMUT, MMAA, MMAB, MMADHC, SUCLG1, SUCLA2, and MCEE), of which 49 were novel. The mut type, caused by the deficiency of methylmalonyl-CoA mutase, was the most common ( n=211, 93%) cause of this condition. c.729_730insTT, c.1106G>A and c.914T>C were the three most frequent mutations in MMUT gene. The frequency of c.914T>C in early-onset patients was significantly higher than that in late-onset patients (8.3% (18/216) vs. 1.6% (1/64), χ 2=3.859, P=0.037). Metabolic crisis was more frequent in mut type than the other types (72.6% (114/157) vs. 3/13, χ 2=13.729, P=0.001),developmental delay and hypotonia were less frequent in mut type (38.2% (60/157) vs. 9/13, 25.5% (40/157) vs. 8/13, χ 2=4.789, 7.705, P=0.030, 0.006). Of the 58 patients identified by newborn screening, 44 patients (75.9%) who were treated from asymptomatic phase developed normally whereas 14 patients (24.1%) who received treatment after developing symptoms exhibited varying degrees of psychomotor retardation. Conclusions:The characteristics of phenotypes and genotypes among Chinese patients with isolated methylmalonic acidemia were analyzed. Expanded the mutation spectrum of the associated genes. Because of the complex clinical manifestations and severe early onset of isolated methylmalonic acidemia, Newborn screening is crucial for early diagnosis and improvement of prognosis. MMUT gene is recommended for carrier screening as an effort to move the test earlier as a part of the primary prevention of birth defects.