[Objective] To analyse and predict the tendencies of using erythrocyte blood in Changsha based on the autoregressive integrated moving average (ARIMA) model, long short-term memory (LSTM) and ARIMA-LSTM combination model, so as to provide reliable basis for designing a feasible and effective blood inventory management strategy. [Methods] The data of erythrocyte usage from hospitals in Changsha between January 2012 and December 2023 were collected, and ARIMA model, LSTM model and ARIMA-LSTM combination model were established. The actual erythrocyte consumption from January to May 2024 were used to assess and verify the prediction effect of the models. The extrapolation prediction accuracy of the models were tested using two evaluation indicators: mean absolute percentage error (MAPE) and root mean square error (RMSE), and then the prediction performance of the model was compared. [Results] The RMSE of LSTM model, optimal model ARIMA(1,1,1)(1,1,1)12 and ARIMA-LSTM combination model were respectively 5 206.66, 3 096.43 and 2 745.75, and the MAPE were 18.78%,11.54% and 9.76% respectively, which indicated that the ARIMA-LSTM combination model was more accurate than the ARIMA model and LSTM model, and the prediction results was basically consistent with the actual situation. [Conclusion] The ARIMA-LSTM model can better predict the clinical erythrocyte consumption in Changsha in the short term.

ObjectiveTo investigate the suppressive effect of 23-hydroxybetulinic acid （23-HBA）， a key constituent of Pulsatillae Radix， on the pulmonary inflammation-related carcinogenesis induced by the combined exposure of 4-（methylnitrosamino）-1-（3-pyridyl）-1-butanone （NNK） and lipopolysaccharide （LPS） in mice， alongside exploring its influence on immune cells and delving into the underlying mechanisms. MethodsA murine model of pulmonary inflammation-related carcinogenesis induced by NNK combined with LPS was established. Mice were randomly assigned into blank control， model， aspirin （10 mg·kg^-1）， and low-， medium-， and high-dose （3.75， 7.5， 15 mg·kg^-1， respectively） 23-HBA groups. The treatment lasted for 26 weeks， after which the spleen， lung， and peripheral blood samples were collected. Lung and spleen indices were calculated. Histopathological changes in the lung tissue were observed by hematoxylin-eosin staining. Immunohistochemistry was employed to assess the expression levels of thyroid transcription factor-1 （TTF-1）， neuron-specific enolase （NSE）， and proliferating cell nuclear antigen （Ki-67） in the lung tissue. High-throughput protein microarray was employed to measure the levels of interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） in the mouse serum. Flow cytometry was employed to evaluate the expression of macrophages， myeloid-derived suppressor cells （MDSCs）， and exhausted T lymphocytes in the lung and spleen tissue. Molecular docking was performed to predict the binding affinity of 23-HBA to Janus kinase 2 （JAK2）， Src homology 2 domain-containing phosphatase 2 （SHP2）， and suppressor of cytokine signaling 3 （SOCS3）. Western blot was performed to assess the protein levels of phosphorylated-signal transducer and activator of transcription 3 （p-STAT3）， p53， and SHP2 in the M1-activated macrophages and A549 lung adenocarcinoma cells treated with 23-HBA. ResultsCompared with the normal group， the lung and spleen indexes of the model group were increased to varying degrees （P<0.05， P<0.01）， the expression of TTF-1， NSE and Ki-67 protein was significantly increased （P<0.05， P<0.01）， and the serum levels of TNF-α， IL-1β and IL-6 were significantly increased （P<0.01）. The number of macrophages in the model group was significantly decreased （P<0.01）， and the number of exhausted T cells and MDSCs was significantly increased （P<0.05， P<0.01）. Compared with the model group， the spleen and thymus index of mice in each dose group of 23-HBA decreased significantly （P<0.05）， and the lung index of mice in the middle dose group of 23-HBA decreased significantly （P<0.05）. The high and middle dose groups of 23-HBA could improve the occurrence of inflammatory infiltration and malignant lesions in the lungs of mice induced by NNK combined with LPS in the model group. The expression of TTF-1 in the middle and high dose groups of 23-HBA was significantly lower than that in the model group （P<0.05， P<0.01）. The expression of NSE and Ki-67 protein in each dose group of 23-HBA was significantly lower than that in the model group （P<0.05， P<0.01）. The contents of IL-1β in the low and high dose groups of 23-HBA were significantly decreased （P<0.05）， and the contents of IL-6 and TNF-α in each dose of 23-HBA were significantly decreased （P<0.05， P<0.01）. The number of macrophages in the lung of the middle dose group of 23-HBA was significantly increased （P<0.05）， and the number of exhausted T cells and MDSCs expressing PD-1 in the lung was significantly decreased （P<0.05， P<0.01）. In addition， 23-HBA had strong molecular docking ability to SHP2， SOCS3 and JAK2 （≥7 kcal·mol^-1）， and significantly down-regulated the protein levels of p-STAT3， SHP2 and p53 in M1 macrophages and A549 lung adenocarcinoma （P<0.01）. Conclusion23-HBA holds promise as a potential therapeutic agent for mitigating pulmonary inflammation and inhibiting malignant transformation induced by the combination of LPS and NNK. It may exert effects by regulating immune cell responses， improving the tumor immune microenvironment， and regulating key signaling pathways.

AIM: To delineate the specific mutation responsible for retinitis pigmentosa(RP)in a Yi pedigree, and to analyze the correlation of RHO gene mutation with clinical phenotype.METHODS:A comprehensive clinical evaluation was conducted on the proband diagnosed with RP and other familial members, complemented by a thorough ophthalmic examination. Peripheral blood samples were obtained from the proband and familial members, from which genomic DNA was extracte. Subsequent whole exome sequencing(WES)was employed to identify the variant genes in the proband. The identified variant gene was validated through Sanger sequencing, then an in-depth analysis of the mutation genes was carried out using genetic databases to ascertain the pathogenic mutation sites. Furthermore, an exhaustive analysis was performed to delineate the genotype and phenotype characteristics.RESULTS:The RP pedigree encompasses 5 generations with 42 members, including 19 males and 23 females. A total of 13 cases of RP were identified, consisting of 4 males and 9 females, which conforms to the autosomal dominant inheritance pattern. The clinical features of this family include an early onset age, rapid progression, and a more severe condition. The patients were found to have night blindness around 6 years old, representing the earliest reported case of night blindness in RP families. The retina was manifested by progressive osteocytoid pigmentation of the fundus, a reduced visual field, and significantly decreased or even vanished a and b amplitudes of ERG. The combined results of WES and Sanger sequencing indicated that the proband had a heterozygous missense mutation of the RHO gene c.1040C>T:p.P347L, where the 1 040 base C of cDNA was replaced by T, causing codon 347 to encode leucine instead of proline. Interestingly, this mutation has not been reported in the Chinese population.CONCLUSION:This study confirmed that the mutant gene of RP in a Yi nationality pedigree was RHO(c.1040C>T). This variant leads to the change of codon 347 from encoding proline to encoding leucine, resulting in a severe clinical phenotype among family members. This study provides a certain molecular, clinical, and genetic basis for genetic counseling and gene diagnosis of RHO.

AIM: To delineate the specific mutation responsible for retinitis pigmentosa(RP)in a Yi pedigree, and to analyze the correlation of RHO gene mutation with clinical phenotype.METHODS:A comprehensive clinical evaluation was conducted on the proband diagnosed with RP and other familial members, complemented by a thorough ophthalmic examination. Peripheral blood samples were obtained from the proband and familial members, from which genomic DNA was extracte. Subsequent whole exome sequencing(WES)was employed to identify the variant genes in the proband. The identified variant gene was validated through Sanger sequencing, then an in-depth analysis of the mutation genes was carried out using genetic databases to ascertain the pathogenic mutation sites. Furthermore, an exhaustive analysis was performed to delineate the genotype and phenotype characteristics.RESULTS:The RP pedigree encompasses 5 generations with 42 members, including 19 males and 23 females. A total of 13 cases of RP were identified, consisting of 4 males and 9 females, which conforms to the autosomal dominant inheritance pattern. The clinical features of this family include an early onset age, rapid progression, and a more severe condition. The patients were found to have night blindness around 6 years old, representing the earliest reported case of night blindness in RP families. The retina was manifested by progressive osteocytoid pigmentation of the fundus, a reduced visual field, and significantly decreased or even vanished a and b amplitudes of ERG. The combined results of WES and Sanger sequencing indicated that the proband had a heterozygous missense mutation of the RHO gene c.1040C>T:p.P347L, where the 1 040 base C of cDNA was replaced by T, causing codon 347 to encode leucine instead of proline. Interestingly, this mutation has not been reported in the Chinese population.CONCLUSION:This study confirmed that the mutant gene of RP in a Yi nationality pedigree was RHO(c.1040C>T). This variant leads to the change of codon 347 from encoding proline to encoding leucine, resulting in a severe clinical phenotype among family members. This study provides a certain molecular, clinical, and genetic basis for genetic counseling and gene diagnosis of RHO.

Objective Logistic regression model was used to analyze the risk factors of liver cancer in patients with hepatitis B virus-related cirrhosis. Methods A retrospective analysis was performed on 504 patients with hepatitis B cirrhosis who were treated in a hospital from April 2021 to April 2024. The occurrence of liver cancer was counted. The risk factors of liver cancer in patients with HBV-related cirrhosis were analyzed by logistic regression analysis. Results Among the 504 patients with hepatitis B cirrhosis, 101 patients developed liver cancer and 403 patients did not develop liver cancer, which were included in the liver cancer group (n=101) and the non-liver cancer group (n=403).. Among hepatitis B cirrhosis, the incidence rate of liver cancer was 20.04%. Compared with the non-liver cancer group, the proportion of patients with long-term drinking history, family history of liver cancer, history of diabetes mellitus, antiviral therapy, and HBV-DNA load>104 were higher in the liver cancer group (P<0.05). logistic regression analysis found that long-term drinking history (OR=3.077, 95%CI: 1.130-8.378, P=0.028), history of diabetes mellitus (OR=3.747, 95%CI: 1.765-7.954, P=0.001), no antiviral therapy (OR=3.466, 95%CI: 1.337-8.985, P=0.011) and HBV-DNA load>104 (OR=3.149, 95%CI: 1.353-7.328, P=0.008) could independently affect the occurrence of liver cancer in patients with hepatitis B cirrhosis. Conclusion According to logistic regression analysis, long-term drinking history, history of diabetes mellitus, no antiviral therapy, and HBV-DNA load>104 are risk factors for liver cancer in patients with HBV-related cirrhosis.

OBJECTIVE To evaluate the influence of pharmaceutical quality control on the efficiency and outcomes of standardized medication therapy management （MTM） services for patients with coronary heart disease by using Economic， Clinical and Humanistic Outcomes （ECHO） model. METHODS This study collected case data of coronary heart disease patients who received MTM services during January-March 2023 （pre-quality control implementation group， n＝96） and June-August 2023 （post-quality control implementation group， n＝164）. Using propensity score matching analysis， 80 patients were selected from each group. The study subsequently compared the economic， clinical， and humanistic outcome indicators of pharmaceutical services between the two matched groups. RESULTS There were no statistically significant differences in baseline data between the two groups after matching （P＞0.05）. Compared with pre-quality control implementation group， the daily treatment cost （16.26 yuan vs. 24.40 yuan， P＜0.001）， cost-effectiveness ratio [23.12 yuan/quality-adjusted life year （QALY） vs. 32.32 yuan/QALY， P＜0.001]， and the incidence of general adverse drug reactions （2.50% vs. 10.00%， P＝0.049） of post-quality control implementation group were decreased significantly； the utility value of the EuroQol Five-Dimensional Questionnaire （0.74± 0.06 vs. 0.71±0.07， P＝0.003）， the reduction in the number of medication related problems （1.0 vs. 0.5， P＜0.001）， the medication adherence score （[ 6.32±0.48） points vs. （6.10±0.37） points， P＝0.001]， and the satisfaction score （[ 92.56±1.52） points vs. （91.95±1.56） points， P＝0.013] all showed significant improvements. Neither group experienced serious adverse drug reactions. There was no statistically significant difference in the incidence of new adverse reactions between the two groups （1.25% vs. 3.75%， P＝0.310）. CONCLUSIONS Pharmaceutical quality control can improve the quality of pharmaceutical care， and the ECHO model can quantitatively evaluate the effect of MTM services， making pharmaceutical care better priced and more adaptable to social needs， thus being worthy of promotion.

BACKGROUND:We have successfully established an animal model of small ear pig in southern Yunnan province with internal tension relieving technique combined with autologous Achilles tendon for anterior cruciate ligament reconstruction,and verified the stability and reliability of the model.However,whether internal tension relieving technique can promote the ligamentalization process of autologous Achilles tendon graft has not been studied. OBJECTIVE:To investigate the differences in the process of ligamentalization between conventional reconstruction and internal reduction reconstruction of the anterior cruciate ligament by gross view,histology and electron microscopy. METHODS:Thirty adult female small ear pigs in southern Yunnan province were selected.Anterior cruciate ligament reconstruction was performed on the left knee joint with the ipsilateral knee Achilles tendon(n=30 in the normal group),and anterior cruciate ligament reconstruction was performed on the right knee joint with the ipsilateral knee Achilles tendon combined with the internal relaxation and enhancement system(n=30 in the relaxation group).The autogenous right forelimb was used as the control group;the anterior cruciate ligament was exposed but not severed or surgically treated.At 12,24,and 48 weeks after surgery,10 animals were sacrificed,respectively.The left and right knee joint specimens were taken for gross morphological observation to evaluate the graft morphology.MAS score was used to evaluate the excellent and good rate of the ligament at each time point.Hematoxylin-eosin staining was used to evaluate the degree of ligament graft vascularization.Collagen fibers and nuclear morphology were observed,and nuclear morphology was scored.Ultrastructural remodeling was evaluated by scanning electron microscopy and transmission electron microscopy. RESULTS AND CONCLUSION:(1)The ligament healing shape of the relaxation group was better at various time points after surgery,and the excellent and good rate of MAS score was higher(P<0.05).Moreover,the relaxation group could obtain higher ligament vascularization score(P<0.05).(2)The arrangement of collagen bundles and fiber bundles in the two groups gradually tended to be orderly,and the transverse fiber connections between collagen gradually increased and thickened,suggesting that the strength and shape degree of the grafts were gradually improved,but the ligament remodeling in the relaxation group was always faster than that in the normal group at various time points after surgery.(3)The diameter,distribution density,and arrangement degree of collagen fibers in the relaxation group were better than those in the normal group at all time points,especially in the comparison of collagen fiber diameter between and within the relaxation group(P<0.05).

Objective To observe the effects of Tongqiao Pingchuan Decoction on miR-155 and NF-κB-HIF-1α signaling pathway in mice with combined allergic rhinitis and asthma syndrome(CARAS);To explore its potential mechanisms in the treatment of chronic inflammation of respiratory tract in CARAS.Methods Totally 60 female BALB/c mice were divided into blank group,model group,miR-155 inhibitor NC group,miR-155 inhibitor group,dexamethasone group and Tongqiao Pingchuan Decoction group using a random number table method,with 10 mice in each group.Except for the blank group,all other groups were sensitized by intraperitoneal injection of ovalbumin combined with aluminum hydroxide gel on days 0 and 7.From days 14 to 30,5%ovalbumin was administered intranasally every other day,followed by continuous nebulization with 5%ovalbumin from days 31 to 37.The miR-155 inhibitor NC group and the miR-155 inhibitor group received intranasal administration of miR-155 inhibitor NC and miR-155 inhibitor solution,respectively,3 hours before nebulization.The blank group was given an equivalent volume of normal saline via intraperitoneal injection,intranasal administration and nebulization.The administration groups were gavaged with the corresponding drug solution 1 hour after nebulization,while the other groups received an equivalent volume of normal saline via gavage.HE staining was used to observe morphology of nasal mucosa and lung tissue,ELISA was used to detect the contents of interleukin(IL)-4,IL-5,and γ-interferon(IFN-γ)in bronchoalveolar lavage fluid,RT-qPCR was used to detect the expression of miR-155,T-bet,GATA-3,NF-κB p65 and HIF-1α mRNA in nasal mucosa and lung tissue,Western blot was used to detect the expressions of T-bet,GATA-3,NF-κB p65 and HIF-1α proteins in lung tissue.Results Compared with the blank group,the model group mice showed shedding of nasal mucosal cilia,tissue congestion and edema,significant glandular hyperplasia and inflammatory cell infiltration,bronchial lumen stenosis,smooth muscle thickening and a large amount of inflammatory substance exudation,the contents of IL-4 and IL-5 increased,while IFN-γ content decreased(P<0.01),the mRNA expression of miR-155,GATA-3,NF-κB p65,HIF-1α in nasal mucosa and lung tissue were increased(P<0.01),while T-bet mRNA expression was decreased(P<0.01),the protein expression of GATA-3,NF-κB p65 and HIF-1α in lung tissue increased(P<0.01),while T-bet protein expression decreased(P<0.01).Compared with the model group,the miR-155 inhibitor group,dexamethasone group and Tongqiao Pingchuan Decoction group showed reduced inflammatory cell infiltration and pathological changes in nasal mucosa and lung tissue of mice,the contents of IL-4 and IL-5 in bronchoalveolar lavage fluid were reduced,while the content of IFN-γ was increased(P<0.01),the expressions of miR-155,GATA-3,NF-κB p65 and HIF-1α mRNA in nasal mucosa and lung tissue were reduced(P<0.05,P<0.01),while the expression of T-bet mRNA increased(P<0.05,P<0.01),the expressions of GATA-3,NF-κB p65 and HIF-1α protein in lung tissue were reduced(P<0.05,P<0.01),the expression of T-bet protein in dexamethasone group and Tongqiao Pingchuan Decoction group mice increased(P<0.05).Conclusion Tongqiao Pingchuan Decoction can effectively inhibit the expression of miR-155,suppress the abnormal activation of NF-κB-HIF-1α signaling pathway,correct the imbalance of Th1/Th2 immunity,and play a therapeutic role in alleviating airway inflammation in CARAS.

Objective To analyze the impact of medical guidance games combined with special-ized nursing on treatment compliance and clinical symptoms in preschool children with pneumonia.Methods A total of 113 preschool children with pneumonia admitted to Beijing Daxing District Peo-ple's Hospital from February 2023 to June 2024 were selected as research subjects and randomly di-vided into control group(n=56)and observation group(n=57).The control group received routine nursing,while the observation group received medical guidance games combined with specialized nursing on the basis of the control group.Treatment compliance,improvement time of clinical symp-toms,and psychological states were compared between the two groups.Results The total treatment compliance rate in the observation group was 98.2％,which was higher than 89.3％in the control group,and the difference was statistically significant(P＜0.05).The disappearance times of symp-toms such as cough,expectoration,dyspnea,and fever in the observation group were all shorter than those in the control group(P＜0.05).After intervention,the scores of the Children's Medical Fear Scale(CMFS)and the Screen for Child Anxiety Related Emotional Disorders(SCARED)in the ob-servation group were both lower than those in the control group(P＜0.05).Conclusion Medical guidance games combined with specialized nursing can improve treatment compliance in preschool children with pneumonia,shorten their recovery time,help alleviate negative emotions such as medi-cal fear and anxiety,and promote disease recovery.

Objective To explore common target genes related to aging and osteoporosis using data from public databases and to screen for small molecule compounds with potential therapeutic effects.Methods Osteoporosis datasets GSE56814 and GSE56815 were downloaded and merged from the public gene expression database(Gene Expression Omnibus(GEO)).Senescent genes were selected and merged from the genecard database and the cell age database.Data cleaning and differential gene selection were performed using R software,and a volcano plot was created.Weighted gene co-expression network analysis was performed to screen out key modules and genes,and the machine learning methods were employed to screen target genes.A Venn diagram was drawn using the the Microbioinformatics platform,and cytoscape software was used to visualise protein interactions results.Interaction analysis of differentially encoded proteins was carried out using the STRING database,and prediction of small molecule compounds was carried out using Cmap database.Finally,Receiver operating characteristic(ROC)curves were plotted to find out the most meaningful target genes,followed by the establishment of a logistic regression model and the creation of a nomogram.Results A total of 127 differential genes were screened using the limma package.Weighted gene co-expression network analysis was performed on the differential genes and 103 highly expressed genes were screened.Three machine learning methods,lasso regression,support vector machine and random forest model,were used to finally obtain four target genes:FOXO3,HIRA,CBX5 and RAD1.The nomogram and calibration curves showed good predictive results.Five small molecule compounds with potential therapeutic effects were identified using the Cmap database:sakuraflavin(O-methylated isoflavone),XMD-885,kangaroo hyoscyamine,everolimus and XMD-1150.Conclusion FOXO3 and HIRA genes have now been confirmed to be the common target genes for aging and osteoporosis,and sakuraflavin has potential therapeutic effects on both conditions.