Objective: To establish an accurate and rapid typing method for Rh typing of samples from patients who have received recent blood transfusions by utilizing the difference in osmotic fragility between fresh and old red blood cells. Methods: A lysing solution suitable for destroying old RBCs was prepared. Sixty-one samples collected in our hospital in 2024 with Rh typing of double groups were treated with the lysing solution to remove the old allogeneic red blood cells while preserving the patient's own fresh red blood cells, followed by repeat Rh typing tests. Results: For 61 samples with Rh typing in double groups, 41 were accurately detected identified through the red blood cell lysis method, yielding an identification rate of 67.21%. No significant difference was observed compared to the detection rate of the commonly used capillary centrifugation modified method (χ =0.103, P>0.05). Conclusion: The red blood cell lysis method provides a novel and rapid experimental approach for clinical use in processing Rh-typed samples that are of double groups, thereby offering a basis for Rh compatibility blood transfusion.

OBJECTIVES: To identify potential molecular targets of quercetin in the treatment of clear cell renal carcinoma (ccRCC). METHODS: The therapeutic targets of quercetin were screened from multiple databases by network pharmacology analysis, and the targets significantly correlated with ccRCC were screened from 4907 plasma proteins using a Mendelian randomization method. The drug-disease network model was constructed to screen the potential key targets. The functions of these targets were evaluated via bioinformatics analysis, and the screened targets were verified in cultured ccRCC cells. RESULTS: Network pharmacology analysis combined with Mendelian randomization identified TP53 (OR=3.325, 95% CI: 1.805-6.124, P=0.0001), ARF4 (OR=0.173, 95% CI: 0.065-0.456, P=0.0003), and DPP4 (OR=0.463, 95% CI: 0.302-0.711, P=0.0004) as the core targets in quercetin treatment of ccRCC. Bioinformatics analysis showed that TP53 was highly expressed in ccRCC, and patients with high TP53 expressions had worse survival outcomes. Molecular docking studies showed that the binding energy between quercetin and TP53 was -5.83 kcal/mol. In cultured 786-O cells, CCK-8 assay and wound healing assay showed that treatment with quercetin significantly inhibited cell proliferation and migration. Quercetin treatment also strongly suppressed the expression of TP53 at both the mRNA and protein levels in 786-O cells as shown by RT-qPCR and Western blotting. CONCLUSIONS TP53 may be the key target of quercetin in the treatment of ccRCC, which sheds light on potential molecular mechanism that mediate the therapeutic effect of quercetin.
Humans ; Quercetin/pharmacology* ; Carcinoma, Renal Cell/genetics* ; Cell Proliferation/drug effects* ; Kidney Neoplasms/genetics* ; Cell Movement/drug effects* ; Tumor Suppressor Protein p53/metabolism* ; Cell Line, Tumor ; Computational Biology

Objective:To prepare a novel ginseng polypeptide thermosensitive hydrogel,and to investigate its repair effect on heat-induced skin injury in the rats and explore the underlying mechanisms.Methods:Thermosensitive hydrogels were formulated using Pluronic F127 and β-sodium glycerophosphate(β-GP),and their phase transition temperatures,spatial structures,elemental compositions,and water retention capacities were evaluated.The rat models of heat-induced skin injury were established and the model rats were divided into PBS group,Gel group,and ginseng polypeptide gel(GP-Gel)group.After 11 d of treatment,the morphological changes of wound and collagen deposition in the wound of the rats in various groups were observed by HE and Masson staining.Immunohistochemistry was used to detect the expression levels of α-smooth muscle actin(α-SMA),connective tissue growth factor(CTGF),basic fibroblast growth factor(bFGF),proliferating cell nuclear antigen(PCNA),cell proliferation marker Ki67,epidermal growth factor(EGF),CD31,vascular endothelial growth factor(VEGF),P50 and P65 proteins in the skin wound tissue of the rats in various groups.Western blotting method was used to detect the expression levels of Toll-like receptor 4(TLR4)in the skin wound tissue of the rats in various groups.ELISA method was used to measure the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-15(IL-15),and interleukin-10(IL-10)in the serum of the rats in various groups.Results:Compared with PBS and Gel groups,the wound area of the rats in GP-Gel group was reduced(P＜0.01),the expression levels of PCNA,Ki67,EGF,CD31,VEGF,α-SMA,and CTGF proteins in the skin wound tissue were increased(P＜0.05 or P＜0.01),and the expression levels of P65 and TLR4 proteins were decreased(P＜0.01);the level of anti-inflammatory factor IL-10 in serum was increased(P＜0.01),while the levels of pro-inflammatory factors TNF-α,IL-1β,IL-6 and IL-15 were decreased(P＜0.05 or P＜0.01).Conclusion:The ginseng polypeptide thermosensitive hydrogel promotes the repair of heat-induced skin injury by enhancing cell proliferation,collagen synthesis,angiogenesis,and reducing inflammatory responses.

Objective To construct a mammography image classification assistant system suitable for Chinese population,and explore the potential of artificial intelligence technology to assist early screening of breast cancer in China.Methods Curated breast imaging subset of digital database for screening mammography(CBIS-DDSM),Mammographic image analysis society database(MIAS)and other international open datasets were used to conduct model training respectively in order to reproduce the mainstream in-depth learning methods in the current literature.The model was also tested on the Chinese breast mammography database(CBMD)provided by Huajiao Technology Co.,Ltd,and the performance was compared.Aiming at the problem that the Chinese population data are not ideal in the performance test of the open dataset training model,an optimization strategy based on the sliding window adjustment mechanism was implemented in combination with the characteristics of Chinese population data.Then a two-stage migration learning method was designed to improve the overall performance of the model,and then development of our system was carried out.Results With the sliding window adjustment mechanism and the CBMD training model after two-stage transfer learning,the accuracy of our developed system was improved from 0.50 of the open datasets to 0.80,precision from 0.54 to 0.82,sensitivity from 0.52 to 0.80,F1 value from 0.52 to 0.80,and AUC value from 0.51 to 0.89 based on the Chinese population dataset as the test set.Conclusion Through the introduction of sliding window adjustment mechanism and two-stage migration learning strategy,the performance of the breast molybdenum target image classification model has been significantly improved in the Chinese population dataset,and our system primarily achieves the purpose of assisting the classification of breast molybdenum target images for the Chinese population.

Objective To conduct an in-depth analysis and feature extraction of the transcriptomics data of 10 types of cancers in order to realize the staging diagnosis of cancer samples.Methods The transcriptomics data of the top 10 cancers having the highest incidence were amassed from the UCSC Xena website,which comprised 4 938 samples and 59 428 genes.With the aid of variational autoencoder,we developed an incremental feature ranking and selection variational autoencoder(IFRSVAE)based on feature importance ranking and incorporating the masking algorithm and the Incremental Feature Selection(IFS).Subsequently,the performance efficiency of our IFRSVAE model was evaluated in conjunction with Random Forest(RF),Support Vector Machine(SVM),and eXtreme Gradient Boosting(XGboost),and it was also compared with other methods.Results Our research extracted 21 features for the ensuing classification.In comparison to the conventional variational autoencoder,recursive feature elimination,and Lasso regression models,the IFRSVAE model attained more favorable performance across all 3 classifiers(highest AUC value,and well performed other indicators).Notably,the IFRSVAE-RF exhibited the most outstanding performance,with an AUC value reaching 85.49%(95%CI:83.24%～87.74%).Moreover,Shapley additive explanations(SHAP)interpretable model illustrated well contributions of the features in our model.Conclusion Our developed IFRSVAE shows certain effectiveness in feature extraction.The constructed IFRSVAE-RF model demonstrates relatively good performance in the task of cancer staging diagnosis,which providing a new and referable idea for research orientation of deep-learning-based diagnostic methods for cancer staging.

Objective:To explore the effects of Xiaoyao Conger Decoction pretreatment on model rats of tinnitus with syndrome of liver-qi stagnation and spleen deficiency (TLSSD) and its preliminary mechanisms.Methods:The rats were divided into four groups: a blank group, a model group, a positive group, and Xiaoyao Conger high- and low-dosage groups, with 7 rats in each group according to random number table method. Except for the blank group, the TLSSD model was prepared by the composite etiological method in all groups of rats. The positive group received a gavage of carbamazepine at 15 mg/kg, while Xiaoyao Conger high- and low-dosage groups were given 30 g/kg and 15 g/kg of Xiaoyao Conger Decoction, respectively. The blank and model groups were administered an equal volume of saline once daily for 4 weeks. The frequency of conditioned avoidance response behaviors in rats was recorded before and at 2 and 4 weeks post-drug administration; TCM syndrome scores were documented before and after treatment; auditory brainstem response (ABR) thresholds were monitored at frequencies of 4, 12, 20, and 28 kHz; IL-1β mRNA expression in the inferior colliculus and cochlea was detected by RT-qPCR; and the levels of γ-aminobutyric acid (GABA) and glutamic acid (Glu) in hippocampal tissues were determined by LC-MS/MS.Results:The model group exhibited symptoms of liver depression, including irritability, yellow fur, and a dark red tongue, along with signs of spleen deficiency such as lethargy, decreased activity, and loose stools. Compared to the model group, rats in the Xiaoyao Conger high- and low-dosage groups demonstrated a reduction in conditioned avoidance response behaviors ( P<0.05 or P<0.01), a decrease in TCM syndrome scores ( P<0.05 or P<0.01), lower ABR thresholds at 12, 20, and 28 kHz ( P<0.01), reduced levels of IL-1β mRNA in the inferior colliculus and cochlea ( P<0.05 or P<0.01), and decreased hippocampal Glu levels ( P<0.01). Conclusion:The therapeutic effect of Xiaoyao Conger Decoction pretreatment on TLSSD rats may be related to its ability to improve the mood disorders and stress, reduce the neuroinflammatory response, regulate the CNS neurotransmitter balance, and correct the overexcitability of the auditory centre, etc.

This paper focuses on extreme environments and systematically analyzes sleep disorders caused by multiple pathogenesis,including circadian rhythm disorder,yin-yang imbalance and qi-blood disorder under weightlessness,emotional depression due to environmental changes,abnormal diet and excretion,and six excesses pathogenic factors,in accordance with the theory of correspondence between nature and human.It proposes harmonizing yin and yang as the core therapeutic principle and guiding framework,with specific methods including calming rebellious qi-blood,regulating qi movement,harmonizing heart and kidney,and dredging and regulating blood vessels,thus forming the"one guiding principle and four specific methods"treatment strategy.Additionally,by integrating the modified application of classic formulas,a diagnostic and therapeutic approach targeting multi-dimensional pathogenesis is established.This study aims to promote the in-depth integration of Traditional Chinese Medicine(TCM)in extreme environmental medicine through TCM theories and innovative application of prescriptions,provide TCM-characterized solutions for health management of workers in extreme environments,and facilitate the transformation of extreme environmental medicine toward the modern medical model of"prevention-treatment integration".

Objective To conduct preliminary evaluation of the reliability and safety of pancreatic duct-to-jejunum stent-bridging internal drainage as a supplementary approach to pancreaticojejunostomy in central pancreatectomy.Methods The clinical data of 28 patients who underwent robotic central pancreatectomy performed by our team between January 2021 and November 2024 were retrospectively collected,and and follow-up of postoperative endocrine and exocrine functions was performed.Based on the methods of digestive tract reconstruction adopted,the patients were divided into a conventional pancreaticojejunostomy group and a pancreatic duct-to-jejunum stent-bridging internal drainage group(PancreaticoJejunal-Stent bridge group).The operative time,digestive tract reconstruction time,and short-term complications were compared between the two groups.Results Among patients undergoing robotic central pancreatectomy,the digestive tract reconstruction time was shorter(t=5.168,P<0.001)in the PancreaticoJejunal-Stent bridge group([31.1±6.3]min)than that in the conventional pancreaticojejunostomy group([49.7±8.9]min)(t=5.168,P<0.001).The total operative time was(172.7±64.6)min in the PancreaticoJejunal-Stent bridge group and(200.1±52.7)min in the conventional pancreaticojejunostomy group,showing no statistically significant difference(t=1.215,P=0.235).In the PancreaticoJejunal-Stent bridge group,one patient developed a postoperative biochemical fistula,and 14 patients developed grade B pancreatic fistulas.Among the 14 patients with grade B pancreatic fistulas,1 case was complicated by fistula-related intra-abdominal infection,and 13 cases had drainage tube retention time of more than 21 days.In the conventional pancreaticojejunostomy group,2 patients developed postoperative biochemical fistulas,and 11 patients developed grade B pancreatic fistulas.Among the 11 patients with grade B pancreatic fistulas,1 case was complicated by fistula-related intra-abdominal infection,and 1 case was complicated by fistula-related intra-abdominal bleeding and infection.No postoperative gastroparesis,pancreatitis,or grade C pancreatic fistulas occurred in either group.There were no statistically significant differences between the two groups in overall postoperative complication rate(P=0.522),postoperative pancreatic fistula rate(P=0.583),intra-abdominal infection rate(P=0.583),or bleeding rate(P=0.464).Conclusion Pancreatic duct-to-jejunum stent-bridging internal drainage optimizes the anastomosis between the distal end of the pancreas and the jejunum during central pancreatectomy,shortens digestive tract reconstruction time,and reduces surgical complexity without increasing the risk of short-term severe postoperative complications.This approach is safe and feasible.

Objective To determine the reference red blood cells with weak agglutination intensity of low positive quali-ty control products by comparing RhD antigen expression intensity difference according to the serological results.Methods The RhD(+)red blood cells were detected by microcolumn gel method with 1 500 times diluted anti-D typing reagent.The samples with weak and strong RhD antigen expression intensity were selected as the reference red blood cells for weak agglu-tination intensity of low positive quality control products,and verification was performed.Results Ten RhD(+)red blood cells were detected with diluted anti-D typing reagent,of which 8 were 1+and 2 were±.Red blood cells with agglutination intensity of 1+were used as the benchmark to determine the maximum dilution ratio of anti-D typing reagent when their ag-glutination intensity was 1+.As the preparation standard of low positive quality control products,the agglutination intensity of red blood cells with low RhD antigen expression intensity was extremely weak±,which was difficult to ensure the stability of its control limit properties.Based on red blood cells with agglutination intensity of±,the maximum dilution ratio of anti-D typing reagent with agglutination intensity of 1+was re-determined as the preparation standard of low positive quality con-trol products,and the results met the requirements of quality control product setting.Conclusion Using red blood cells with low RhD antigen expression intensity as the benchmark to set the weak agglutination intensity of the low positive quality control products can avoid the loss of control due to the low target value.

Objective: To investigate the function and potential mechanism of hepatocellular carcinoma(HCC) cells-derived extracellular nicotinamide phosphoribosyl transferase(eNAMPT) in the angiogenesis of human umbilical vein endothelial cells(HUVEC). Methods: The eNAMPT secretion of HCC cells were stimulated by hypoxia treatment and investigated the angiogenesis of HUVEC was investigated by tube formation assays. Transcriptomic RNA sequencing(RNA-seq) analyses of HUVEC under eNAMPT treatment were used to explore the mechanism of the effect of eNAMPT-insulin receptor(INSR) axis on angiogenesis. Western blot was used to detect the effects of eNAMPT-INSR axis on protein expression of HUVEC cells. qRT-PCR was used to detect the influence of eNAMPT-INSR axis on the expression of HUVEC intracellular related target genes. Results: ELISA and Western blot results showed that the protein level of eNAMPT in the culture supernatant of HCC cells increased significantly after hypoxia stimulation.In vitroangiogenesis assay results showed that eNAMPT significantly increased the nodules number and total length of tubes in HUVEC. The transcriptome sequencing results of HUVEC showed that eNAMPT, as an insulin analogue, participated in the regulation of biological processes such as cell migration and angiogenesis in HUVEC, and might promote angiogenesis by activating insulin-like signaling pathways. Western blot results showed that the eNAMPT-INSR axis affected p-AKT and p-ERK1/2 protein levels in PI3K-AKT and MAPK-ERK1/2 signaling pathways in HUVEC. qRT-PCR results showed that the eNAMPT-INSR axis affected the expression of angiogenesis related target genes MMP2, MMP9 and VEGF in HUVEC. Conclusion eNAMPT secreted by HCC cells promotes angiogenesis by binding INSR to activate the PI3K-AKT and MAPK-ERK1/2 signaling pathways.