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 provide a reference for the implementation and high-quality development of hospital medication reconciliation. METHODS A semi-structured questionnaire was designed to investigate the implementation of drug reconciliation services in medical institutions before and after the release of 5 standards such as Standard for Medication Reconciliation Services in Medical Institutions（“standards” for short，in 2021 and 2022）. Descriptive statistical analysis was conducted on the survey results. RESULTS After the promulgation of the standards， the medication reconciliation service rate of all types of medical institutions increased from 15.10% （434/2 874） in 2021 to 27.84%（363/1 304） in 2022. In 2022， in the 363 medical institutions providing drug reconciliation services， the median number of pharmacists involved in drug reconciliation was 6. The participation rate of pharmacists in standardized training for drug reconciliation services was 75.00%， among which the participation rate of third-class hospitals was higher， reaching 85.71%. The main stages covered by medication reconciliation services included patient admission， transfer between departments， and discharge. The main problems found in the service included repeated medication （252， 69.42%）， inappropriate usage and dosage （228， 62.81%）， drug interactions and adverse reactions （218， E-mail：cputianxin@163.com 60.06%）. Only 69 institutions （19.01%） had a separate electronic information recording system， while 48 institutions 58516003。E-mail：zhenjiancun@vip.163.com （13.22%） had established comprehensive quality management and evaluation improvement systems. In terms of value embodiment， 141 institutions （38.84%） did not provide any form of compensation to relevant pharmacists. “Closely linked to enhancing patient satisfaction and improving services” was the most significant experience influencing medication reconciliation work（192， 52.89%）， while “the shortage of talent which meet the relevant requirements” stands as the primary challenge faced by medical institutions at all levels（238， 65.56%）. CONCLUSIONS The release of the standards has effectively improved the development rate of medication reconciliation in national medical institutions. However， there is still room for improvement in various aspects， including the allocation of personnel for medication reconciliation services， service content， information management， and the construction of quality control and evaluation systems.

ObjectiveUltra-performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS/MS） was used to investigate the metabolism and distribution of nardosinone in rats， then metabolic pathways were speculated. MethodRats were administered with 30 mg·kg^-1 of nardosinone suspension by gavage for 3 consecutive days， and plasma， urine， feces， and tissues of heart， liver， spleen， lung， kidney， brain， stomach， and intestine were collected at predetermined time points. After treatment， the samples were processed for UPLC-Q-Exactive Orbitrap MS/MS， and the MS data were analyzed using Xcalibur 2.2 software. The metabolites were searched by comparing the base peak chromatogram and extracted ion chromatogram between the treated group and blank group， and based on the relative retention time（t_R）， quasi-molecular ion peak， precise molecular mass， and fragment ions of MS/MS， the elemental composition were searched using databases such as SciFinder and PubChem， as well as referring to relevant literature， the possible metabolites were identified and the metabolic pathways were inferred. ResultA total of 30 metabolites of nardosinone were identified， including 15， 19， 12， 7， 4， 11， 8， 13， 13， 8 and 12 metabolites in urine， feces， plasma， brain， heart， liver， spleen， lung， kidney， stomach and intestine， respectively. The main metabolic pathways of nardosinone in rats were hydroxylation， dehydroxylation， reduction， dehydrogenation， hydration， dehydration， carboxylation， glucuronidation， and dehydroxy-isopropyl. ConclusionNardosinone can be metabolized by phase Ⅰ and phase Ⅱ metabolism in rats， and the metabolites are widely distributed in the major organs. The results of this study can provide a basis for further research on the pharmacodynamic material basis， pharmacological mechanism and clinical application of nardosinone.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Bovine coronavirus(BCoV)can cause diarrhea in calves,as well as respiratory diseases,and lead to the death of calves in severe cases,resulting in great economic losses to the cattle pro-duction.BCoV is highly prevalent and widely spread.So far in China there is no independently de-veloped vaccine against BCoV on the market.In this study,the most conserved N gene of BCoV was first selected to construct a prokaryotic expression vector,and the N protein was expressed and purified;subsequently,BALB/c mice were immunized with BCoV N protein combined with Freund's adjuvant,and the antibody potency,the proportion of splenic T-lymphocyte subpopulation and the cytokine release of the immunized mice were detected by ELISA and flow cytometry.The results showed that a soluble BCoV N protein about 55 kDa in size was successfully obtained.The ELISA test showed that the antibody potency of serum IgG and IgA was 1∶51 200 and 1∶3 200,respectively,in mice immunized with BCoV N protein combined with Freund's adjuvant.The flow cytometry test showed that the percentage of CD4+/CD8+T-lymphocytc subsets was extremely higher in the immunized mice(P＜0.01),and the release of TNF-α was significantly higher(P＜0.05),producing a cellular immune response biased towards the Th1 type in comparison with that in control mice.It is indicated that the soluble expression of BCoV N protein can be successfully a-chieved by the prokaryotic expression system,and the obtained BCoV N protein presents good im-munogenicity which induces strong humoral,mucosal and cellular immune responses in the immu-nized mice.Our study provides an important technical support for the development of a safe and ef-fective BCoV subunit vaccine.

Objective:To analyze the clinical effect of Jiawei Shengxian decoction in the treatment of acquired weakness in patients with mechanical ventilation in intensive care unit (ICU).Methods:A prospective study method was used to select 68 patients with severe mechanical ventilation who met the diagnostic criteria of ICU acquired weakness (ICU-AW) and were admitted to the Department of Critical Care Medicine, Nanchong Hospital of Traditional Chinese Medicine from May 2022 to May 2023 as the study objects, and were divided into observation group and control group according to random number table method, with 34 cases in each group. The control group was treated with conventional Western medicine treatment and rehabilitation treatment, and the observation group was treated with Jiawei Shengxian decoction based on syndrome differentiation of the control group. The muscle strength indexes, symptom scores, inflammatory factors, recovery time, serum biochemical indexes and safety indexes of the two groups were evaluated and compared before and after treatment.Results:Before treatment, there was no statistically significant difference in muscle strength indicators, symptom scores, inflammatory factors, and serum biochemical indicators between the two groups (all P>0.05). After treatment, the Medical Research Council (MRC) muscle strength score, diaphragm thickness and diaphragm motion of observation group were higher than those of control group (all P<0.05). After treatment, the scores of Acute Physiology and Chronic Health Evaluation Ⅱ (APACHEⅡ), multiple organ failure (MODS) and traditional Chinese medicine syndrome in the observation group were lower than those in the control group, and the Barthel index (BI) score of activities of daily living in the observation group was higher than that in the control group (all P<0.05). After treatment, the serum levels of procalcitonin (PCT), hypersensitive C-reactive protein (hs-CRP) and white blood cell count (WBC) in the observation group were lower than those in the control group (all P<0.05). The mechanical ventilation time, ICU admission time, getting out of bed activity time and hospital stay time of the observation group were shorter than those of the control group (all P<0.05). After treatment, the levels of hemoglobin (HGB) and platelet count (PLT) in the observation group were higher than those in the control group, and the levels of serum total bilirubin (TBIL), blood urea nitrogen (BUN) and serum creatinine (Scr) in the observation group were lower than those in control group (all P<0.05). Conclusions:Jiawei Shengxian decoction has a good effect in the adjuvant treatment of acquired asthenia in ICU patients with mechanical ventilation, which can enhance the muscle strength of limbs and the activity ability of diaphragm, improve the symptom score of patients, reduce the expression level of inflammatory factors, shorten the rehabilitation process of patients, and improve the serum biochemical indexes of patients.

Objective:To investigate the efficacy and mechanism of modified Yinjia formula in the treatment of persistent genital tract chlamydia trachomatis infections based on the targeted regulation of CD8+T cell autophagy pathway.Methods:Thirty-six BALB/c mice,except for the blank group,were inoculated intravaginally with chlamydia to establish a model of persistent chlamydia trachomatis infection.After successful modeling,patients were divided into groups and given low,medium and high doses of Yinjia modifiedformula and azithromycin by continuous gavage for 4 weeks,respectively.HE staining was used to observe the histomorphologic changes of uterus and fallopian tube tissues of the mice.Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression level of chlamydia trachomatis genes in the low reproductive tract.Proportion of OX40 and LAG-3 positive cells in mouse lymph nodes was detected by flow cytometry.Immunoblotting was used to detect the expression of autophagy-related protein Beclin-1,Atg5,LC3Ⅱ/LC3Ⅰ,p62,and Ras/Raf/MEK/ERK pathway protein in mouse splenic CD8+T cells.Results:Compared with the model group,the inflammatory cell infiltration was reduced to different degrees in all treatment groups.Among them,only a very small amount of in-flammatory cell infiltration was observed in the uterus and fallopian tubes in the high-dose group of traditional Chi-nese medicine,and no significant inflammatory exudation and thickening was observed in the fallopian tubes.Compared with the model group,the expression of Beclin-1,Atg5,LC3Ⅱ/LC3Ⅰ was decreased in all treatment groups(P＜0.01),while the protein expression of p62,Ras,P-Raf,P-MEK and P-ERK was up-regulated(P＜0.01).Additionally,the proportion of OX40 and LAG-3 positive cells in lymphocytes was decreased(P＜0.01).A-mong them,Beclin-1,Atg5,LC3Ⅱ/LC3Ⅰ protein expression as well as the percentage of OX40 and LAG-3 positive cells were the lowest in the high-dose group of traditional Chinese medicine(P＜0.01).Conclusions:The modi-fied Yinjia formula may inhibit the autophagic activity of CD8+T cells through the Ras/Raf/MEK/ERK signaling pathway,thereby enhancing the targeted clearance function of chlamydia trachomatis inclusion bodies by CD8+T cells and delaying the process of persistent chlamydia trachomatis infection in the reproductive tract.

Objective:To investigate the efficacy and mechanism of modified Yinjia formula in the treatment of persistent genital tract chlamydia trachomatis infections based on the targeted regulation of CD8+T cell autophagy pathway.Methods:Thirty-six BALB/c mice,except for the blank group,were inoculated intravaginally with chlamydia to establish a model of persistent chlamydia trachomatis infection.After successful modeling,patients were divided into groups and given low,medium and high doses of Yinjia modifiedformula and azithromycin by continuous gavage for 4 weeks,respectively.HE staining was used to observe the histomorphologic changes of uterus and fallopian tube tissues of the mice.Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression level of chlamydia trachomatis genes in the low reproductive tract.Proportion of OX40 and LAG-3 positive cells in mouse lymph nodes was detected by flow cytometry.Immunoblotting was used to detect the expression of autophagy-related protein Beclin-1,Atg5,LC3Ⅱ/LC3Ⅰ,p62,and Ras/Raf/MEK/ERK pathway protein in mouse splenic CD8+T cells.Results:Compared with the model group,the inflammatory cell infiltration was reduced to different degrees in all treatment groups.Among them,only a very small amount of in-flammatory cell infiltration was observed in the uterus and fallopian tubes in the high-dose group of traditional Chi-nese medicine,and no significant inflammatory exudation and thickening was observed in the fallopian tubes.Compared with the model group,the expression of Beclin-1,Atg5,LC3Ⅱ/LC3Ⅰ was decreased in all treatment groups(P＜0.01),while the protein expression of p62,Ras,P-Raf,P-MEK and P-ERK was up-regulated(P＜0.01).Additionally,the proportion of OX40 and LAG-3 positive cells in lymphocytes was decreased(P＜0.01).A-mong them,Beclin-1,Atg5,LC3Ⅱ/LC3Ⅰ protein expression as well as the percentage of OX40 and LAG-3 positive cells were the lowest in the high-dose group of traditional Chinese medicine(P＜0.01).Conclusions:The modi-fied Yinjia formula may inhibit the autophagic activity of CD8+T cells through the Ras/Raf/MEK/ERK signaling pathway,thereby enhancing the targeted clearance function of chlamydia trachomatis inclusion bodies by CD8+T cells and delaying the process of persistent chlamydia trachomatis infection in the reproductive tract.

Bovine coronavirus(BCoV)can cause diarrhea in calves,as well as respiratory diseases,and lead to the death of calves in severe cases,resulting in great economic losses to the cattle pro-duction.BCoV is highly prevalent and widely spread.So far in China there is no independently de-veloped vaccine against BCoV on the market.In this study,the most conserved N gene of BCoV was first selected to construct a prokaryotic expression vector,and the N protein was expressed and purified;subsequently,BALB/c mice were immunized with BCoV N protein combined with Freund's adjuvant,and the antibody potency,the proportion of splenic T-lymphocyte subpopulation and the cytokine release of the immunized mice were detected by ELISA and flow cytometry.The results showed that a soluble BCoV N protein about 55 kDa in size was successfully obtained.The ELISA test showed that the antibody potency of serum IgG and IgA was 1∶51 200 and 1∶3 200,respectively,in mice immunized with BCoV N protein combined with Freund's adjuvant.The flow cytometry test showed that the percentage of CD4+/CD8+T-lymphocytc subsets was extremely higher in the immunized mice(P＜0.01),and the release of TNF-α was significantly higher(P＜0.05),producing a cellular immune response biased towards the Th1 type in comparison with that in control mice.It is indicated that the soluble expression of BCoV N protein can be successfully a-chieved by the prokaryotic expression system,and the obtained BCoV N protein presents good im-munogenicity which induces strong humoral,mucosal and cellular immune responses in the immu-nized mice.Our study provides an important technical support for the development of a safe and ef-fective BCoV subunit vaccine.

Objective:To analyze the clinical effect of Jiawei Shengxian decoction in the treatment of acquired weakness in patients with mechanical ventilation in intensive care unit (ICU).Methods:A prospective study method was used to select 68 patients with severe mechanical ventilation who met the diagnostic criteria of ICU acquired weakness (ICU-AW) and were admitted to the Department of Critical Care Medicine, Nanchong Hospital of Traditional Chinese Medicine from May 2022 to May 2023 as the study objects, and were divided into observation group and control group according to random number table method, with 34 cases in each group. The control group was treated with conventional Western medicine treatment and rehabilitation treatment, and the observation group was treated with Jiawei Shengxian decoction based on syndrome differentiation of the control group. The muscle strength indexes, symptom scores, inflammatory factors, recovery time, serum biochemical indexes and safety indexes of the two groups were evaluated and compared before and after treatment.Results:Before treatment, there was no statistically significant difference in muscle strength indicators, symptom scores, inflammatory factors, and serum biochemical indicators between the two groups (all P>0.05). After treatment, the Medical Research Council (MRC) muscle strength score, diaphragm thickness and diaphragm motion of observation group were higher than those of control group (all P<0.05). After treatment, the scores of Acute Physiology and Chronic Health Evaluation Ⅱ (APACHEⅡ), multiple organ failure (MODS) and traditional Chinese medicine syndrome in the observation group were lower than those in the control group, and the Barthel index (BI) score of activities of daily living in the observation group was higher than that in the control group (all P<0.05). After treatment, the serum levels of procalcitonin (PCT), hypersensitive C-reactive protein (hs-CRP) and white blood cell count (WBC) in the observation group were lower than those in the control group (all P<0.05). The mechanical ventilation time, ICU admission time, getting out of bed activity time and hospital stay time of the observation group were shorter than those of the control group (all P<0.05). After treatment, the levels of hemoglobin (HGB) and platelet count (PLT) in the observation group were higher than those in the control group, and the levels of serum total bilirubin (TBIL), blood urea nitrogen (BUN) and serum creatinine (Scr) in the observation group were lower than those in control group (all P<0.05). Conclusions:Jiawei Shengxian decoction has a good effect in the adjuvant treatment of acquired asthenia in ICU patients with mechanical ventilation, which can enhance the muscle strength of limbs and the activity ability of diaphragm, improve the symptom score of patients, reduce the expression level of inflammatory factors, shorten the rehabilitation process of patients, and improve the serum biochemical indexes of patients.