[Objective] To investigate the factors associated with alanine aminotransferase (ALT) abnormalities in multi-ethnic blood donors across five Zang autonomous prefectures in the plateau regions of Qinghai Province, and to provide evidence for ensuring blood safety and formulating screening strategies. [Methods] A retrospective analysis was performed on the ALT abnormal test results of blood donors in the Zang autonomous prefectures of Qinghai from 2022 to 2024. The correlations between ALT levels and factors including gender, age, altitude, and infectious markers were investigated. [Results] The overall ALT unqualified rate among blood donors in this region was 9.01%. Significant differences in ALT levels were observed across genders and age groups (P<0.05). Variations in ALT abnormality rates were also noted among different plateau regions (P<0.05). Overall, ALT values exhibited an increasing trend with rising altitude. The average ALT unqualified rates were 11.19% in Zang donors, 7.96% in Han donors, and 4.79% in donors from other ethnic groups (P<0.05). No statistically significant association was observed between ALT abnormality and the presence of HBV/HCV infectious markers (P>0.05). [Conclusion] In the plateau areas of Qinghai, multi-ethnic blood donors have a relatively high ALT levels and ALT unqualified rates, showing distinct regional characteristics. ALT elevation in voluntary blood donors is related to non-pathological factors such as gender, age, and dietary habits, but not to infectious indicators.

Objective To evaluate the disease risk of Vibrio parahaemolyticus (VP) in aquatic products of raw food in Guangzhou. Methods VP detection was carried out in aquatic products of raw food sold in Guangzhou from 2009 to 2022. Gene sequence and wgSNP analysis of 30 VP strains (including 15 food strains and 15 patient strains) were performed for the detection rate of pathogenic VP. sQMRA was applied to assess VP risk of aquatic products of raw food. Results The detection rate of VP in raw aquatic products in Guangzhou was 7.30% (98/1 343). The detection rate of TDH virulence gene in patient strains was 86.70% (13/15) , and the detection rate of TRH was 6.67% (1/15). In 15 food strains, TDH and TRH were negative. The WgSNP analysis showed that 2 food strains had high similarity with the patient strains, indicating the same cluster. Risk assessment showed that the number of Vibrio parahaemolyticus infection cases caused by intaking aquatic products of raw food in Guangzhou was 384 ever year. Conclusion The detection rate of VP in aquatic products of raw food is high in Guangzhou , and the detection rate of VP virulence genes in aquatic products of raw food is low. Gene sequence and wgSNP analysis can be used for risk assessment of food pathogenic bacteria. The risk of disease of Vibrio parahaemolyticus in aquatic products of raw food is high.

OBJECTIVE To investigate the effects of metformin （Met） on liver injury in non-alcoholic steatohepatitis （NASH） rats by regulating the PI3K/AKT/PDGF signaling pathway. METHODS NASH model was constructed by feeding rats with a high- glucose and high-fat diet， and assigned into Model group， Met low-dose group （Met-L group， 100 mg/kg）， Met medium-dose group （Met-M group， 200 mg/kg）， Met high-dose group （Met-H group， 400 mg/kg）， and high dose of Met+PI3K activator group （Met-H+740 Y-P group， 400 mg/kg Met+50 mg/kg 740 Y-P）， with 12 rats in each group. Another 12 rats were regarded as the Control group. Each group of rats was orally administered/injected with the corresponding medication once a day for 6 consecutive weeks. The changes in body weight and liver index of rats were recorded and analyzed. The pathological damage [evaluation of non-alcoholic fatty liver disease activity score （NAS）]， lipid deposition （calculation of the proportion of oil red O-positive staining area）， and fibrosis （calculation of collagen deposition score） were observed in liver tissue of rats. The levels of inflammatory factors [interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α）] in serum and liver tissue， the levels of serum lipid metabolism indicators [total cholesterol （TC）， triglyceride （TG）， and low-density lipoprotein cholesterol （LDL-C）] and liver function indicators [aspartate aminotransferase （AST） and alanine Δ 基金项目 武汉市知识创新专项项目（No.2022020801010588）； aminotransferase （ALT）] were measured. The expression levels of PI3K/AKT/PDGF signaling pathway-related proteins and Caspase-3 in liver tissue of rats were determined. RESULTS Compared with the Control group， body weight， liver index， the levels of serum lipid metabolism indicators and liver function indicators， the levels of IL-6 and TNF-α in serum and liver tissue， the NAS， the proportion of oil red O-positive staining area， the collagen deposition fraction， and the levels of phosphorylated PI3K and AKT proteins， as well as the expression levels of PDGF and Caspase-3 proteins in liver tissue， were all significantly increased （P＜0.05）. The liver tissue showed severe pathological damage， characterized by an abundance of lipid droplets and pronounced collagen deposition. After the intervention with Met， the aforementioned quantitative indicators and pathological changes in rats were significantly improved in a dose- dependent manner （P＜0.05）. 740 Y-P could reverse the improvement effects of high dose of Met on the above indexes of rats （P＜ 0.05）. CONCLUSIONS Met can improve liver damage， and alleviate inflammatory reactions and liver fibrosis of NASH rats， the mechanism of which may be associated with inhibiting PI3K/AKT/PDGF signaling pathway.

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.

Wilson's disease （WD） is a copper metabolism disorder caused by mutations in the ATP7B gene， with diverse phenotypes and complex pathogenesis. It is one of the few rare diseases that can achieve good clinical efficacy through standardized treatment. Since there are few systematic reviews of this disease， we summarize the pathogenesis and treatment methods of WD from traditional Chinese and western medicine by reviewing the literature related to WD. In western medicine， ATP7B gene mutation is considered as the root cause of WD， which affects copper transport and causes copper metabolism disorders. The excessive copper deposited in the body will result in oxidative stress， defects in mitochondrial function， and cell death. Western medicine treatment of WD relies mainly on drugs， and copper antagonists are the first choice in clinical practice， which are often combined with hepatoprotective and antioxidant therapy. Surgery is a common therapy for the patients with end-stage WD， and gene therapy provides an option for WD patients. According to the traditional Chinese medicine （TCM） theory， WD is rooted in constitutional deficiency and copper accumulation and triggered by dampness-heat accumulation or phlegm combined with stasis. The patient syndrome varies in different stages of the disease， and thus the treatment should be based on syndrome differentiation. The TCM treatment method of nourishing the liver and kidneys and warming the spleen and kidneys can address the root cause. The methods of clearing heat and drying dampness， resolving phlegm and dispelling stasis， and soothing liver and regulating qi movement can be adopted to treat symptoms. On the basis of syndrome differentiation， special prescriptions for the treatment of WD have been formulated， such as Gandou decoction， Gandouling， and Gandou Fumu decoction， which have been widely used in clinical practice. TCM and western medicine have their own advantages and shortcomings. The integrated Chinese and western medicine complementing with each other demonstrates great therapeutic potential. This paper summarizes the pathogenesis and treatment of WD with integrated Chinese and western medicine， aiming to provide a reference for the clinical diagnosis and treatment of this disease.

Objective To investigate the application effects of different myocardial protective solutions in total thoracoscopic minimally invasive aortic valve replacement surgery.Methods The clinical data of 72 patients with total thoracoscopic minimally invasive aortic valve replacement surgery in this hospital from May 2020 to January 2024 were analyzed retrospectively.The patients were divided into the St Thomas cardioplegia group(STH group,n=13),del Nido cardioplegia group(DN group,n=24),histidine tryptophan ketoglutar-ate solution group(HTK group,n=35)according to the different myocardial protective solutions.The levels of lactate(Lac)before and during surgery,the highest levels of myocardial creatine kinase isoenzyme(CK-MB),high-sensitivity troponin T(TnT)and creatinine(Cr)before operation,on the operative day and after surgery as well as the duration of extracorporeal circulation,aortic cross-clamping time,maximum flow rate,minimum bladder temperature,cardioplegia perfusion times,number of defibrillation after aortic de-clamping,postoperative ventilator assisted time,ICU stay duration and postoperative hospitalization duration were com-pared among the three groups.Results Except for 1 case of HTK was discharged automatically after surgery,the other 71 cases recovered and discharged according to the doctor's advice.There were no statistically signif-icant differences in the age,body weight,extracorporeal circulation time,aortic blocking time,maximum flow volume of extracorporeal circulation,minimum bladder temperature of extracorporeal circulation,Lac before extracorporeal circulation,highest Lac during extracorporeal circulation,assistant time of postoperative venti-lator,ICU stay duration,postoperative hospitalization duration,serum Cr before operation,Cr on operative day,preoperative TnT,postoperative TnT on operative day,postoperative highest TnT,preoperative CK-MB,postoperative CK-MB on operative day and postoperative highest CK-MB among the three groups(P＞0.05).There were statistically significant differences in the defibrillation ratio after aortic de-clamping and perfusion frequency of myocardial protective solution(P＜0.05).There was statistically significant difference in the perfusion frequency of myocardial protective solution in pairwise comparison among groups(P＜0.05),and the defibrillation ratio after aortic de-clamping had statistical difference between the DN group and HTK group(P＜0.05).Conclusion DN,STH and HTK all have good myocardial protective effect in total thoraco-scopic minimally invasive aortic valve surgery.HTK has the advantages of less perfusion times and decreasing the operative procedures compared with DN and STH;DN has the advantage of lower use for electrical defib-rillation correcting arrhythmias after aortic opening over HTK.

Objective To evaluate the accuracy and clinical application value of the fluorescence quantitative PCR melting curve meth-od for detecting fungal nucleic acid.Methods 460 suspected or confirmed patients with respiratory fungal infections were enrolled in the study.The fluorescence quantitative PCR melting curve method was used as the test method,and the fungal 26S rRNA gene nucleic acid detection kit combined with Sanger sequencing was used as the reference method.Sputum samples from each study subject were collected and detected by the test method and reference method,respectively.The Kappa value of the two methods was calculated to evaluate the consistency of the results.Results Compared with the reference method,the overall conformity rate of the test method was 92.83%(427/460).Compared with the reference method,the positive conformity rates,negative conformity rates,and overall conformity rates of the test method for detecting 8 fungi,including Candida albicans,Candida glabrata,Candida krusei,Candida trop-icalis,Candida parapsilosis,Cryptococcus neoformans,Candida guilliermondii,and Aspergillus,were 97.34%(183/188),97.06%(264/272),and 97.17%(447/460),100.00%(33/33),99.77%(426/427),and 99.78%(459/460),100.00%(16/16),99.55%(442/444),and 99.57%(458/460),98.11%(52/53),99.75%(442/444),and 99.57%(458/460),95.08%(58/61),99.50%(397/399),and 98.91%(455/460),100.00%(9/9),99.56%(449/451),and 99.57%(458/460),85.00%(17/20),99.32%(437/440),and 98.70%(454/460),and 97.59%(81/83),97.88%(369/377),and 97.83%(450/460),respectively.The Kappa values for the consistency evaluation of the two methods'detection results were both greater than 0.8.Upon retesting the inconsistent re-sults of the two methods,it was found that 53.7%(22/41)of the detection results were consistent with the test method,and the others were consistent with the reference method.Conclusion The fluorescence quantitative PCR melting curve method can simultaneously detect 8 kinds of fungi,and the detection results are highly consistent with the reference method.It has unique advantages in fungal de-tection and important clinical application value.

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.