[Objective] To investigate the factors influencing the detection of HLA-C expression by flow cytometry. [Methods] A total of 12 hematopoietic stem cell suspension samples from peripheral hematopoietic stem cell volunteer donors were randomly collected after CD34⁺ cell counting detection. The influence of detecting different number of nucleated cell (500 000, 50 000 and 5 000), sequential order of red blood cell lysis and antibody incubation, and the HLA-C antibody with varied remaining time from the expiration date on the detection results of HLA-C expression by flow cytometry were investigated, respectively. The significance of differences between different groups was analyzed through Student t test. [Results] There was no significant difference in the proportion of HLA-C positive cells and mean fluorescence intensity (MFI) among the three groups with different nucleated cell numbers detected (500 000, 50 000 and 5 000) (P>0.05). The sequential order of red blood cell lysis and antibody incubation had no influence on the proportion of HLA-C positive cells (P>0.05), but HLA-C MFI value was significantly lower when antibody incubation was performed after red blood cell lysis than that when antibody incubation was performed before red blood cell lysis (P<0.05). The proportion of HLA-C positive cells and MFI value detected by HLA-C antibody remaining 24 months from the expiration date were significantly higher than those detected by HLA-C antibody remaining only 5 months from the expiration date (P<0.05). [Conclusion] The present study has investigated the factors of influencing HLA-C expression level by flow cytometry, the results have important reference and application value for standardizing the experimental operation of HLA-C expression and improving the accuracy and comparability of detection results.

ObjectiveThis study aims to explore the mechanism of action of Huangqi Guizhi Wuwutang in treating rheumatoid arthritis （RA）-associated sarcopenia by regulating autophagy and improving skeletal muscle homeostasis based on network pharmacology，bioinformatics，machine learning，and animal experiments. MethodsActive ingredients and targets of Huangqi Guizhi Wuwutang were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP），PubChem，and SwissTargetPrediction databases. RA-related datasets were retrieved from the GEO database，and differential genes were screened. Sarcopenia-related targets were searched through GeneCards and the Comparative Toxicology Database （CTD），and autophagy-related gene sets were downloaded from the Human Autophagy Database （HADb）. Their intersection was analyzed to identify autophagy-related therapeutic targets，followed by enrichment analysis. A protein-protein interaction （PPI） network was constructed using the STRING database，and key targets were selected using multiple methods. Machine learning was applied to predict models based on the expression profiles of intersecting targets，and nomogram models were constructed based on key targets. Molecular docking of the top four active ingredients with key targets was performed using AutoDockVina. A collagen-induced arthritis （CIA） rat model was established using bovine type Ⅱ collagen，with SD rats divided into groups including a blank group，a model group，and low-，medium-，and high-dose groups of Huangqi Guizhi Wuwutang （2.44，4.88，and 9.76 g·kg^-1） and administered for five consecutive weeks. Joint scores and gastrocnemius muscle mass were recorded and analyzed after modeling. Hematoxylin and eosin （HE） staining and Masson's staining were used to observe pathological changes in muscle tissue. Immunofluorescence staining was applied to observe the protein expression levels of myosin heavy chain （MYHC） and insulin-like growth factor-1 （IGF-1） in skeletal muscle. Western blot was used to detect the protein expression levels of autophagy-related proteins ATG5，Beclin1，LC3B，muscle-specific proteins （MuRF1），MaFbx，and MYHC. Real-time quantitative reverse transcription PCR （Real-time PCR） was performed to measure the mRNA expression levels of ATG5，Beclin1，LC3B，MuRF1，MaFbx，and MYHC in muscle tissue. ResultsNetwork pharmacology revealed that Huangqi Guizhi Wuwutang shared 25 common targets with autophagy genes related to RA-associated sarcopenia. The PPI network and machine learning identified six key targets，which were primarily involved in autophagy and inflammatory pathways. Animal experiments showed that compared to the blank group，the model group had significantly higher joint scores （P<0.01） and lower gastrocnemius muscle index （P<0.01）. HE staining indicated a significant reduction in the cross-sectional area of gastrocnemius muscle fibers，with notable inflammatory cell infiltration and muscle atrophy in the model group. Masson's staining revealed obvious collagen fiber proliferation and deposition，with significant muscle fibrosis in the model group. The protein and mRNA expression levels of ATG5，Beclin1，LC3B，MuRF1，and MaFbx were significantly increased （P<0.01），while the protein expression of MYHC and IGF1 was significantly downregulated （P<0.01）. Compared with the model group，the high-dose group of Huangqi Guizhi Wuwutang showed significantly reduced protein and mRNA expression levels of ATG5，Beclin1，LC3B，MuRF1，and MaFbx （P<0.01） and increased protein expression levels of MYHC and IGF1 （P<0.01）. The cross-sectional area of muscle fibers increased，and the muscle cell morphology approached normal. Moreover，pathological abnormalities in the gastrocnemius muscle were significantly improved，with reduced collagen fiber proliferation （P<0.01）. ConclusionHuangqi Guizhi Wuwutang can mediate autophagy by regulating the expression of ATG5，Beclin1，LC3B，and IGF1，thereby reducing skeletal muscle catabolism and improving skeletal muscle homeostasis，which contributes to the treatment of RA-associated sarcopenia. The findings provide insight into the mechanisms underlying the effects of Huangqi Guizhi Wuwutang in the treatment of RA-related sarcopenia and offer a reference for its enhanced clinical application.

Objective: To establish a "regular blood donor red blood cell gene database"(hereafter referred to as the "database") by applying molecular biology techniques for red blood cell antigens genotyping and utilizing information technology software, and to determine the significance and application value of this "database" in precise red blood cell transfusion. Methods: Fifteen antigens [C, c, E, e, M, N, S, s, Fy (a), Fy (b), Jk (a), Jk (b), Le (a), Le (b), P1] across six blood group systems (RHCE, MNS, FY, JK, Lewis and P1PK) were detected among 9 426 regular blood donors using the TaqMan-MGB method combined with an improved U-shaped microplate approach. With the assistance of information technology software, the "database" was integrated into the overall inventory management system of the blood supply chain. This enabled comprehensive management of regular blood donor and patient information, test results, specific antigen screening for regular blood donors, graded antigen matching between donors and patients, and rare blood type donor records. Results: The TaqMan-MGB method successfully detected paired antigens (C/c, E/e, M/N, S/s, Fy /Fy , Jk /Jk ) within a single reaction well using a standardized PCR amplification protocol. This method provided a reliable testing solution for clinical institutions and empowered blood collection and supply organizations with high-throughput screening capabilities. In the blood supply chain, genotyped red blood cells accounted for 13.2% (721/5 462 U) of the total inventory, with 95.34% (348/365) originating from donors who donated two units of blood. Moreover, the “database” fulfilled 94.06% (443/471 U) of compatible transfusion requirements from medical institutions and effectively managed rare blood type donors. Conclusion: The establishment of the "database" facilitated the transition of blood compatibility testing from traditional serological methods to molecular biology-based gold standard techniques, significantly advancing the implementation of precise transfusion strategies based on multi-antigen matching between donors and patients.

Cough variant asthma is a distinct subtype of asthma characterized by chronic irritant dry cough as the sole or predominant clinical manifestation. It is one of the primary causes of chronic cough in children. In traditional Chinese medicine, it belongs to the category of " spasmodic cough", " wind-induced cough", " chronic cough", and " asthmatic cough". At present, Western medicine treatment approaches mainly focus on symptomatic treatment, but fail to fully deal with its complex systemic mechanisms, and have limitations such as poor control of clinical symptoms and rebound exacerbation upon treatment cessation. Based on the theory of " systemic qi stagnation", this paper proposes that the core pathogenesis of cough variant asthma in children is qi stagnation, intermingling of phlegm and blood stasis, and obstruction of collaterals. Disease progression is fundamentally driven by exogenous pathogen activation of endogenous predispositions, particularly dysregulation of sanjiao qi movement, which serves as the primary disease-inducing factor. During the acute phase, the treatment principle focuses on dispelling wind and ventilating lung to restore physiological qi. As the disease progresses to the progressive phase, the focus shifts toward smoothing liver and purging lung to resolve qi counterflow. In the chronic phase, therapeutic strategy prioritizes dissipating phlegm and eliminating blood stasis to smooth collaterals. Finally, during the remission phase, treatment emphasizes strengthening spleen and kidney to consolidate the foundation and cultivate the vitality. This integrative approach synergizes the external elimination of latent pathogens, internal harmonization of qi movement, and dredging collaterals by dispelling blood stasis. It also incorporates the theory of " gentle dispersion to expel pathogens and moistening dryness to harmonize collaterals", aiming to provide a theoretical basis and effective prescriptions for the integrated traditional Chinese and Western medicine treatment of cough variant asthma in children.

Objective:To explore the mediating effect of psychological resilience between structural empowerment and work engagement of clinical nurses.Methods:This study was a cross-sectional study. Using the convenient sampling method, a total of 1 723 nurses from three Class Ⅲ Grade A hospitals in Henan Province were selected as the research objects from March to May 2023. The survey was conducted using general information questionnaire, Conditions of Work Effectiveness Questionnaire-Ⅱ (CWEQ-Ⅱ), Nurses Psychological Resilience Scale and Utrecht Work Engagement Scale (UWES). Pearson correlation analysis was used to explore the correlation between structural empowerment, psychological resilience and job engagement. AMOS 24.0 software was used to establish the structural equation model and analyze the mediating effect.Results:A total of 1 723 questionnaires were sent out in this study, and 1 590 were effectively collected, with an effective recovery rate of 92.28%. The total scores of structural empowerment, psychological resilience and job engagement of 1 590 clinical nurses were (70.42±15.61), (96.84±13.15) and (62.24±11.02), respectively. Pearson correlation analysis showed that job engagement was positively correlated with structural empowerment and psychological resilience ( r=0.637, 0.670, P<0.01). The results of structural equation model showed that psychological resilience had a partial mediating effect between structural empowerment and job engagement. The mediating effect value was 0.321, accounting for 48.86% of the total effect. Conclusions:In this study, the structural empowerment, psychological resilience and job engagement of clinical nurses are above the medium level, and psychological resilience has a partial mediating effect between structural empowerment and job engagement. Nursing managers should fully empower nurses at the organizational level, take targeted measures to improve their psychological resilience and enhance their level of work engagement.

Objective To explore the value of the Kyoto classification score in differentiating undifferentiated gastric cancer from differentiated gastric cancer,and establish a predictive scoring system for differentiating undifferentiated gastric cancer under endoscope.Methods 183 gastric cancer patients were retrospectively analyzed.According to pathology,95 patients were included in the differentiated group and 88 were included in the undifferentiated group.The age,gender and Kyoto classification score of patients in the two groups were compared,and the factors associated with undifferentiated gastric cancer were screened by binary Logistic regression analysis.The predictive scoring system for undifferentiated gastric cancer was established based on the obtained odds ratio(O(R))values,and the receiver operator characteristic curve(ROC curve)was drawn.Results Compared with differentiated group,the total scores of Kyoto classification,atrophy,intestinal metaplasia and diffuse redness were lower in undifferentiated group(P<0.01).Under the age of 55(P<0.05),female(P<0.05),and C1 atrophy or no atrophy(P<0.01)were independently associated with undifferentiated gastric cancer.The area under the curve(AUC)of predictive scoring system for undifferentiated gastric cancer was 0.881(95％CI:0.828～0.934),and the sensitivity and specificity were 80.70％and 90.50％at the optimal cut-off value.Conclusion There are differences in Kyoto classification scores between undifferentiated and differentiated gastric cancer patients.The predictive scoring system of undifferentiated gastric cancer established by us has certain value in distinguishing undifferentiated gastric cancer under endoscope.

Objective To observe the effect of quercetin on mechanical allodynia,astrocyte activation,and upregulation of pain-related transient receptor potential vanilloid 1(TRPV1)and P2X purinoceptor 3(P2X3)in mice with paclitaxel-induced peripheral neuropathy.Methods Twenty-four C57BL/6 mice were divided randomly into control,model,and model+quercetin groups(n=8 mice per group).Paclitaxel(total dose 8 mg/kg)was injected intraperitoneally into mice in the model and model+quercetin groups to establish the model.Mice in the control group were injected intraperitoneally with the same volume of vehicle.On day 8 after the first injection,mice in the model+quercetin group were injected with 60 mg/kg quercetin solution orally and mice in the other groups were injected with the same volume of vehicle.Mechanical pain was measured by the von Frey test.Activation of astrocytes in the spinal dorsal horn was detected by immunofluorescence.Expression levels of TRPV1 and P2X3 in dorsal root ganglia were detected by immunofluorescence and Western Blot.Results(1)Compared with model group,the mechanical pain of mice in model+quercetin group were relieved.(2)Compared with model group,the activation of astrocytes and the expressions of TRPV1 and P2X3 in mice of model+quercetin group were alleviated(P＜0.05).Conclusions Quercetin can significantly reduce mechanical pain in mice with paclitaxel-induced peripheral neuropathy.This mechanism maybe related to alleviating the activation of astrocytes in the spinal dorsal horn and reducing expression of TRPV1 and P2X3 in the dorsal root ganglia.

Pulmonary lymphangiomyomatosis(LAM)is a rare chronic progressive diffuse cystic lung disease that mainly occurs in women of reproductive age.Pulmonary hypertension is a rare complication of LAM.Currently,there is insufficient evidence on the epidemiology,pathogenesis and treatment strategy of LAM related pulmonary hypertension.We reported a case of a woman at reproductive age with shortness of breath and diagnosed with LAM by the combination of specific lung imaging features and serum vascular endothelial growth factor D.Precapillary pulmonary hypertension was confirmed by right cardiac catheterization.Her condition was stable with Sirolimus and home oxygen therapy.

Pulmonary hypertension is a rare complication of pneumoconiosis,which is caused by long term denture dusk contacting and poor protection.Here we reported a case,who was a denture technician and had been engaged in denture grinding for more than 10 years.According to the specific lung imaging findings and dust exposure history,she was diagnosed with interstitial lung disease and pneumoconiosis (probable).Precapillary pulmonary hypertension was confirmed by right cardiac catheterization.

Objective To establish the energy expenditure(EE)prediction models of health Qigong Wuqinxi based on heart rate combined with accelerometer counts in different body parts,so to provide a reference for monitoring EE of Wuqinxi.Methods Seventy-four healthy college students aged 18-30 were selected as the research objects.They were divided into a skilled group of 39(21 males and 18 females)and a primary group of 35(17 males and 18 females)according to their level of practice,with 55 in the model group and 19 in the validation group.When performing a set of Wuqinxi,all subjects were recorded heart rate(HR),accelerometer counts[the average counts of X-axis,Y-axis,Z-axis and vector magnitude(VM)],and EE-related indicators by wearing the Polar heart rate moni-tor,ActiGraph-GT3X+accelerometers(9 parts:waist,both arms,both wrists,both thighs,and both ankles)and CORTEX Meta Max3B-R2 portable indirect calorimeter.Based on the accurate EE mea-sured by the gas metabolism meter,linear regression models of only HR,only accelerometer counts,or HR combined accelerometer counts were established,and their accuracy was analyzed and com-pared.Results The EE of Wuqinxi was significantly correlated with gender,height,weight,muscle mass,HR-related indicators,and accelerometer counts in different parts(X-axis counts of the waist,VM values of the left thigh and counts of the X-axis and Y-axis,VM values of the right thigh and the Y-axis,Z-axis count value,left ankle VM value and Y-axis count value,right ankle VM value and Y-axis,Z-axis count value)(P<0.05).Moreover,the adjusted R2 of the prediction model of only HR,only accelerometer counts,and the both were 0.582,0.508 and 0.678,respectively(P<0.05).The correlation between the predicted and measured values ranged between 0.706 and 0.817.Accord-ing to the Bland-Altman analysis,for each model,all except one error value fell outside the 95%confidence interval.That is,all models had an excellent fitting effect and high accuracy.Among them,the combined model was of the best prediction effect:EE(kcal)=-20.089+0.279×body weight(kg)+0.243×ΔHR(exercise HR-quiet HR,bpm)+0.001×Right thigh Y-axis count value+0.181×exer-cise HR(bpm)-4.202×gender(male=0,female=1).Conclusion The EE prediction model of Wuqinxi es-tablished on HR combined with accelerometer counts has the best effect and can calculate the EE more accurately.