Objective: To retrospectively analyze the prevalence of hepatitis C virus (HCV) infection among voluntary blood donors in Qinghai Province over a ten-year period and to provide evidence for refining blood safety screening strategies. Methods: Blood samples from 362 066 blood donors in Qinghai collected between January 2015 and April 2024 were simultaneously screened using enzyme-linked immunosorbent assay (ELISA) and nucleic acid testing (NAT). Follow-up was conducted for donors with reactive HCV RNA screening results, and alanine transaminase (ALT) was detected by rate method. Results: The HCV positive rate among blood donors in Qinghai was 0.22%. Gender, marital status, number of blood donations, and educational level were associated with HCV infection. Significant differences in HCV positive rates were observed among donors across regions and ethnic groups. The HCV positive rate among donors in Golog Tibetan Autonomous Prefecture (with an average altitude of 4 330 m) was significantly higher than that in Xining (0.52% vs 0.21%, P<0.001). Positivity rates were also significantly higher in Salar (0.84%), Hui (0.81%), Zang (0.60%), and Tu (0.45%) ethnic groups compared to the Han ethnic group (0.17%) (P<0.001). The abnormal rate of ALT in HCV-positive donors was higher than in non-HCV donors (6.13% vs 1.55%) (P<0.001). Conclusion: The relatively high HCV positivity rate among blood donors in Qinghai highlights the need for further investigation into viral sources, risk factors, and transmission routes. Optimized screening strategies are essential to ensure blood safety.

Objective: To genotype 50 RhD variant samples from Guangzhou, China, using our previously established genotyping strategy, thereby providing guidance for transfusion management and antenatal monitoring in RhD-variant individuals. Methods: Between June and August 2024, fifty samples identified as RhD variants during RhD-negative confirmation testing at Guangzhou Blood Center were collected. Serological testing for the D antigen was performed with two different anti-D reagents, and the epitope profiles of the D antigen were determined using a commercial panel of monoclonal anti-D reagents containing nine kinds of monoclonal anti-D. Genomic DNA was extracted, and high-resolution melting (HRM) analysis was applied to detect the Asian-type DEL (RHD 1227A). Subsequently, RHD genotyping was carried out using Multiplex Ligation-dependent Probe Amplification (MLPA) and Sanger sequencing. Results: Among the 50 D variant samples, 17 (34.0%) Asian type DEL samples were detected by HRM, including 13 cases with RHD DEL1/01N.01 genotype and 4 cases with RHD DEL1/DEL1 genotype. Eleven (11/50, 22.0%) samples were typed as DVI by the epitope profiles of D antigen. The epitope profiles of D antigen combined with Sanger sequencing of exon 6 identified 5 (5/50, 10.0%) cases of RHD weak partial 15/01N.01. MLPA combined with Sanger sequencing identified two cases of RHD DVI.3/DEL1, representing 4.0% (2/50) of the samples. Additionally, the following RHD genotypes were each detected in one case: RHD weak D type 18/01N.04, RHD weak D type 72/01N.01, RHD weak D type 95/DEL1, RHD weak D type 114/DEL1, RHD weak D type 136/DEL1, RHD weak D type 147/01N.01, RHD 496G/496G, RHD 536C/01N.01, RHD 689A/689A, RHD 689A/DEL1, RHD DEL32/DEL1, RHD DV.1/01N.01, RHD DV.5/01N.01, RHD 01.01/01N.01, and RHD 01/01N.01. Conclusion: Fifty D variant individuals were typed using our previously established serological and molecular approach. These findings provide guidance for precision transfusion therapy in RhD variant patients and inform evidence-based decisions regarding anti-D immunoglobulin prophylaxis for RhD variant pregnant women.

Objective:To analyze the relationship between the expression level of calumenin(CALU)and the prognosis of hepatocellular carcinoma(HCC)patients by bioinformatics tools and establish the prognostic prediction nomogram,and to clarify its possible mechanism.Methods:The data of 374 HCC tissue samples were downloaded from The Cancer Genome Atlas(TCGA)database and the data of 160 normal tissue samples were down loaded from Genotype-Tissue Expression(GTEx)database.Paired sample t-test was used to analyze the difference in CALU expression between the HCC tissue samples and the paired adjacent normal tissue samples.Human Protein Atlas(HPA)database was used to verify the results.DESeq2 package was used to screen the differentially expressed genes(DEGs)between CALU low expression group and CALU high expression group in the HCC tissue samples.R package pROC was used to analyze the receiver operating characteristic(ROC)curve.Univariate and multivariate Cox regression analyses were used to confirm the prognosis value of CALU in the HCC patients with different clinicopathological characteristics,and ggplot2 package was used to construct the forest plot.R packages rms and survival were used to construct the nomogram and its calibration curve,and the diagnostic value of CALU in distinguishing HCC tissue from normal tissue was analyzed.The data from Kaplan-Meier Plotter database were used to further verify the relationship between CALU and the prognosis of HCC patients.The gene transcriptional expression data of 216 HCC samples obtained from GSE14520 dataset were used to verify the prediction accuracy of the nomogram.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were used to determine the function and enrichment pathways of the DEGs,and Gene Set Enrichment Analysis(GSEA)was used to obtain the significantly enriched gene sets of the DEGs.Single-cell sequencing data of 10 HCC tissue samples and 8 adjacent normal tissue samples obtained from GSE149614 dataset were used to verify the relationship between CALU and the prognosis of HCC patients and its mechanism.Results:Compared with normal tissue,the expression level of CALU mRNA in HCC tissue was significantly increased(P＜0.001),and the expression level of CALU protein in HCC samples was also increased.A total of 928 DEGs were identified between CALU low expression group and CALU high expression group in the HCC samples,including 784 upregulated DEGs and 144 downregulated DEGs.The ROC analysis results indicated that CALU showed high diagnostic value in distinguishing cancer tissue from adjacent non-cancer tissue with an area under curve(AUC)of 0.839.Kaplan-Meier survival analysis showed that the survival rate of HCC patients in CALU high expression group was significantly lower than that in CALU group low expression(P＜0.001).Univariate and multivariate Cox regression analyses results demonstrated that high expression of CALU was an independent risk factor of the prognosis in HCC patients,and a prognosis prediction nomogram was constructed.The applicability of nomogram on the prognosis of HCC was verified by GSE14520 dataset.The GO enrichment analysis results showed that DEGs were mainly enriched in pathways related to the oxidative stress,ferroptosis and cuproptosis(P＜0.05).The KEGG enrichment analysis results showed that DEGs were mainly enriched in the pathways related to extracellular matrix(ECM)receptor interaction,linoleic acid metabolism and neuroactive ligand receptor interaction(P＜0.05).The GSEA results showed that high expression of CALU may promote the G1-S phase transition of the cell cycle,ubiquitination protein polymerization and HCC progression,while low expression of CALU may activate oxidative stress,ferroptosis and cuproptosis in HCC cells.Single-cell sequencing analysis results showed that the expression level of CALU mRNA was significantly increased in HCC cells with advanced tumor stages.HCC_CALU_High cell subset was mainly related to ubiquitination,p53 and cell cycle(P＜0.01),and HCC_CALU_Low cell subset was mainly related to oxidative stress,ferroptosis,and histone binding(P＜0.01).Conclusion:The high expression of CALU may be related to the poor prognosis of HCC patients.The constructed nomogram of HCC prognosis shows favourable effect in predicting the survival rate of the HCC patients.The up-regulation of CALU may promote HCC progression by regulating the G1-S phase of the cell cycle and ubiquitination of protein,while down-regulation of CALU may inhibit HCC progression by inducing oxidative stress,ferroptosis and cuproptosis in cells.

Objective:To investigate the changes of pathogenic bacteria distribution and immune response of urinary tract infection(UTI)after pyeloureteroplasty in children.Methods:A total of 150 children with obstructive hydronephroplasty who received pyeloureteroplasty in Qinghai Women and Children's Hospital from February 2020 to June 2023 were selected as the study objects.The postoperative urine of the children was collected,the pathogens were identified by automatic bacterial analyzer,and the patients were divided into infected group and uninfected group according to the infection status.The detection of pathogenic bacteria in children with UTI was analyzed.The clinical data of the two groups were compared,and the influencing factors of postoperative UTI were screened by Logistic regression analysis,and the prediction model was established and verified.The changes of immune response were compared between the two groups.Results:UTI occurred in 56 cases(37.33%)of 150 children who underwent pyeloureteroplasty.There were 58 strains of gram-negative bacteria,29 strains of gram-positive bacteria and 2 strains of fungi isolated.The main pathogenic bacteria was Escherichia coli.Multivariate Logistic regression analysis showed that serum creatinine(SCr)and hydronephrosis volume/renal volume(HV/RV)levels increased,urinary catheter retention time,urinary bag replacement time and hospital stay were risk factors for UTI after pyeloureteroplasty in children,increased age and elevated albumin(Alb)were protective factors(P<0.05).Through Bootstrap self-sampling,the prediction model had good differentiation and accuracy.There was no significant difference in the levels of CD4+,CD8+and IgM between the two groups before surgery(P>0.05),and the levels of CD4+,CD8+and IgM were signi-ficantly improved between the two groups after surgery,and the improvement degree of the uninfected group was significantly better than that of the infected group(P<0.05).Conclusion:The pathogens of UTI in children after pyeloureteroplasty are mainly gram-nega-tive bacteria,and the immune function is increased 7 days after the operation.

Objective:To investigate the changes of pathogenic bacteria distribution and immune response of urinary tract infection(UTI)after pyeloureteroplasty in children.Methods:A total of 150 children with obstructive hydronephroplasty who received pyeloureteroplasty in Qinghai Women and Children's Hospital from February 2020 to June 2023 were selected as the study objects.The postoperative urine of the children was collected,the pathogens were identified by automatic bacterial analyzer,and the patients were divided into infected group and uninfected group according to the infection status.The detection of pathogenic bacteria in children with UTI was analyzed.The clinical data of the two groups were compared,and the influencing factors of postoperative UTI were screened by Logistic regression analysis,and the prediction model was established and verified.The changes of immune response were compared between the two groups.Results:UTI occurred in 56 cases(37.33%)of 150 children who underwent pyeloureteroplasty.There were 58 strains of gram-negative bacteria,29 strains of gram-positive bacteria and 2 strains of fungi isolated.The main pathogenic bacteria was Escherichia coli.Multivariate Logistic regression analysis showed that serum creatinine(SCr)and hydronephrosis volume/renal volume(HV/RV)levels increased,urinary catheter retention time,urinary bag replacement time and hospital stay were risk factors for UTI after pyeloureteroplasty in children,increased age and elevated albumin(Alb)were protective factors(P<0.05).Through Bootstrap self-sampling,the prediction model had good differentiation and accuracy.There was no significant difference in the levels of CD4+,CD8+and IgM between the two groups before surgery(P>0.05),and the levels of CD4+,CD8+and IgM were signi-ficantly improved between the two groups after surgery,and the improvement degree of the uninfected group was significantly better than that of the infected group(P<0.05).Conclusion:The pathogens of UTI in children after pyeloureteroplasty are mainly gram-nega-tive bacteria,and the immune function is increased 7 days after the operation.

Breast cancer is the most common cancer in women and one of the deadliest cancers worldwide.According to the distribution of tumor tissue,breast cancer can be divided into invasive and non-invasive forms.The cancer cells in invasive breast cancer pass through the breast and through the immune system or systemic circulation to different parts of the body,forming metastatic breast cancer.Drug resistance and distant metastasis are the main causes of death from breast cancer.Research on breast cancer has attracted extensive attention from researchers.In vitro construction of tumor models by tissue engineering methods is a common tool for studying cancer mechanisms and anticancer drug screening.The tumor microenvironment consists of cancer cells and various types of stromal cells,including fibroblasts,endothelial cells,mesenchymal cells,and immune cells embedded in the extracellular matrix.The extracellular matrix contains fibrin proteins(such as types Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅵ,and Ⅹcollagen and elastin)and glycoproteins(such as proteoglycan,laminin,and fibronectin),which are involved in cell signaling and binding of growth factors.The current traditional two-dimensional(2D)tumor models are limited by the growth environment and often cannot accurately reproduce the heterogeneity and complexity of tumor tissues in vivo.Therefore,in recent years,research on three-dimensional(3D)tumor models has gradually increased,especially 3D bioprinting models with high precision and repeatability.Compared with a 2D model,the 3D environment can better simulate the complex extracellular matrix components and structures in the tumor microenvironment.Three-dimensional models are often used as a bridge between 2D cellular level experiments and animal experiments.Acellular matrix,gelatin,sodium alginate,and other natural materials are widely used in the construction of tumor models because of their excellent biocompatibility and non-immune rejection.Here,we review various natural scaffold materials and construction methods involved in 3D tissue-engineered tumor models,as a reference for research in the field of breast cancer.

Objective To explore the differences in brain functions between individuals with good acclimatization to high altitudes and those with poor acclimatization based on microstate and functional connectivity analysis of resting-state electroencephalogram(EEG)activities.Methods A total of 44 residents exposed to the high-altitude environment of 3 650 m above sea level were enrolled.They were divided into two groups according to their findings for the high-altitude acclimatization index(AAI),with 22 in the good acclimatization group and 22 in the poor acclimatization group.The good acclimatization group had 10 males and 12 females of the mean age of(20.48±2.09)years.Among them,10 were Tibetans,and 12 were Han Chinese.The poor acclimatization group consisted of 18 males and 4 females,with a mean age of(18.90±1.25)years.Among them,7 were Tibetans,and 15 were Han Chinese.The K-means clustering algorithm was used to classify EEG microstates into four categories of A,B,C,and D.The average duration,coverage,occurrence frequency,and transition probabilities between microstates were compared.Coherence analysis(COH)was performed to assess the strength of brain network connectivity.Results Compared to the good acclimatization group,the poor acclimatization group exhibited a longer duration,higher occurrence frequency,and coverage of microstate C.The coverage of microstate D was also higher,while the occurrence frequency and coverage of microstate B were lower.The poor acclimatization group showed a decrease in transitions from microstate B to A or D,while transitions from C to B or D,and those from D to A or C increased,showing statistically significant differences between the groups(P＜0.05).In the α,β,δ,and θ frequency bands,the poor acclimatization group had higher COH strength in regions such as the frontal and occipital lobes,showing symmetrical functional connectivity between the left and right hemispheres.Conclusion The good and poor high-altitude acclimatization groups exhibit different neural mechanisms in terms of microstates and functional connectivity.There are significant differences between individuals with good acclimatization and those with poor acclimatization regarding visual information processing,interference resistance,brain network stability,and coherence.

Objective To observe the changes in the symptoms and relevant physiological indicators in subjects after inhaling the hypoxic air produced by a hypoxic air generator at a low altitude prior to their entry into high-altitude environment,and to explore its early warning effect for acute mountain sickness(AMS)among the subjects upon their subsequent entry into high-altitude environment.Methods A total of 50 subjects who were going to visit high-altitude regions were enrolled.All subjects were men,with an average age of(22.00±1.52)years.They continuously inhaled for 30 minutes hypoxic air(which simulated the air at the altitude of 5200 m,with an oxygen content 10.80％)generated by a hypoxic air generator.During this period fingertip oxygen saturation,heart rate,blood pressure,and symptoms of discomfort were observed and recorded.On the fourth day after living at an altitude of 4020 m,the subjects completed the evaluation for the symptom scores of acute mild altitude disease(AMAD).The subjects were divided into a maladjusted group(18 cases)and a well-adjusted group(32 cases)according to whether they experienced discomfort(including drowsiness,dizziness,chest tightness,cold sweating of the hands,etc.)during the inhalation of hypoxic air at a low altitude.After entry into the high-altitude environment,they were divided into an AMS group(28 cases)and a non-AMS group(22 cases)according to whether they experienced AMS after entering the he high-altitude environment.The primary indicator was the incidence of AMS,including the incidence of AMAD and severe acute mountain sickness(SAMS),and the incidence of AMS in the maladjusted group and the well-adjusted group after entering high-altitude environment.The secondary indicator was the relationship between the changes in fingertip oxygen saturation after inhaling hypoxic air at a low altitude and the incidence of AMS and the AMAD symptom scores.Results All 50 subjects traveled by air to the target altitude of 4020 m above sea level at the same time.The AMS incidence among them was 56.0％(28/50),with the incidence of AMAD being 54.0％(27/50)and the incidence of SAMS being 2.0％(1/50).In the single case of SAMS,the patient had high-altitude pulmonary edema.The incidences of AMS after entering high-altitude environment in the maladjusted and well-adjusted groups were 88.9％(16/18)and 37.5％(12/32),respectively,and the difference was statistically significant(P＜0.01).In the 50 subjects,fingertip oxygen saturation decreased rapidly in the first 11 minutes into the inhalation of hypoxic air at a low altitude,with a more pronounced decrease in the AMS group than that in the non-AMS group,and the differences between the groups were statistically significant after 5,9,and 11 minutes(P＜0.05).Fingertip oxygen saturation plateaued in the 50 subjects from the 12th to the 30th minute,with no significant differences between the AMS and non-AMS groups.The mean value of fingertip oxygen saturation within 30 minutes of hypoxic air inhalation was negatively correlated with the AMAD symptom scores after subjects'entry into high-altitude environment(r=-0.300).Conclusion Those who experience symptoms of discomfort after exposure to hypoxic air produced by a hypoxic air generator at a low altitude are more likely to develop AMS and close attention should be paid to the decrease in fingertip oxygen saturation within the first 11 minutes.

{L-End}Objective To investigate the current status of long working hours of food-delivery workers from food-delivery platform, and analyze its impact on their health such as occupational stress, depressive symptoms, insomnia symptoms, fatigue accumulation, and life satisfaction. {L-End}Methods A total of 2 145 food-delivery workers from two food-delivery platforms in Guangzhou City were selected as the research subjects using convenience sampling method. The occupational stress, depressive symptoms, insomnia symptoms, fatigue accumulation, and life satisfaction were investigated using the Core Occupational Stress Scale, Patient Health Questionnaire-9 items, Self-sleep Management Questionnaire, Self-diagnosis Questionnaire of Fatigue Accumulation of Workers, and World Health Organization Five-item Well Being Index. {L-End}Results The median and the 25th and 75th percentiles of weekly working hours were 63 (49,77) hours. And 92.2% (1 978/2 145) of workers, who worked more than 40 hours per week, were long working hours workers. While 70.1% (1 504/2 145) workers, who worked 55 hours or more per week, were ultra-long working hours workers. The detection rate of occupational stress, depressive symptoms, insomnia symptoms, fatigue accumulation, and low life satisfaction was 30.1%, 27.5%, 34.7%, 40.8%, and 75.1%, respectively. The longer the weekly working hours of the research subjects, the higher the detection rate of occupational stress and fatigue accumulation (all P<0.01). The results of multivariate logistic regression analysis showed that ultra-long working hours (weekly working hours ≥55 hours) was the influencing factor of occupational stress and fatigue accumulation after excluding the influence of confounding factors among workers (all P<0.05). {L-End}Conclusion The food-delivery workers of food-delivery platforms generally work long hours. Ultra-long working hours is a risk factor for occupational stress and fatigue accumulation among these workers.

Objective:To explore the influencing and prognosis factors of emphysematous urinary tract infection (EUTI).Methods:The baseline clinical data of the patients admitted to Shandong University Qilu Hospital (Qingdao) from December 2013 to June 2020 and diagnosed with EUTI were analyzed retrospectively. The patients with non-EUTI (NEUTI) during the same period were selected as the control group. The baseline characteristics between the two groups were compared. Logistic regression analysis method was used to analyze the influencing factors of EUTI.Results:(1) 24 EUTI patients and 53 NEUTI patients were included in the present study. Compared with the NEUTI group, the hemoglobin level was lower ( t=-5.245, P<0.001) and the levels of blood urine nitrogen ( Z=-4.361, P<0.001), serum creatinine (Scr, Z=-4.543, P<0.001), blood glucose ( Z=-2.608, P=0.009), and triacylglycerol ( Z=-2.408, P=0.016) were higher in the EUTI group. The proportions of diabetes mellitus ( χ2=13.453, P<0.001) and chronic kidney disease ( χ2=17.936, P<0.001) in the EUTI group were higher than those in the NEUTI group. Increasing Scr was the risk factor of EUTI in patients with urinary tract infection ( OR=1.011, 95% CI 1.001-1.020, P=0.025). (2) Escherichia coli ( E.coli, 14 cases, 58.3%) was the most common causative organism. The other causative organisms included Klebsiella pneumoniae (2 cases, 8.3%), Enterococcus faecium (1 case, 4.2%), Pantoea (1 case, 4.2%), and mixed bacteria of E.coli and Enterococcus faecium (1 case, 4.2%). Ten cases of E.coli were extended-spectrum β-lactamases (ESBL)-positive. (3) Of the 24 patients with EUTI, 4 patients had adverse outcomes. The length of stay ( Z=-2.457, P=0.014), blood urea nitrogen ( t=2.432, P=0.024), shock ( P=0.002), autoimmune disease ( P=0.022), and white blood cell count ( Z=-2.091, P=0.036) were statistically different between good prognosis group ( n=20) and poor prognosis group ( n=4). However, logistic regression analysis results showed that neither was the influencing factor of poor prognosis of EUTI. Conclusions:The elevated Scr level is the independent influencing factor of EUTI among urinary infection patients. E.coli is the most common pathogenic bacteria, and ESBL-positive bacteria are common.