OBJECTIVE To explore the correlation between drug consumption index and diagnosis related groups （DRG） overspending cases， and provide a basis for hospitals to optimize the cost structure and strengthen the refined management. METHODS Based on the data of DRG patients enrolled in a third-grade class A hospital from September to November 2023， the multivariate Logistic regression model and restricted cubic spline （RCS） model were used to analyze the correlation of drug consumption index with DRG overspending cases and its threshold effect， respectively. At the same time， rational drug use evaluation was conducted based on the drug consumption index， precise cost control and management were carried out， and the changes in the main pharmaceutical indicators of the whole hospital were analyzed before control （January-June 2023） and after control （January-June 2024）. RESULTS The results of multivariate Logistic regression analysis showed that long hospitalization days， high drug consumption index， transfer to other departments and combined diabetes mellitus were the risk factors for DRG overspending （P＜0.05）. The results of the RCS model showed that the drug consumption index had a non-linear relationship with DRG overspending. When the drug consumption index was ≥0.64， the drug consumption index was positively correlated with the risk of DRG overspending（P＜0.05）. Compared with the same period before the control， medical cost per time， drug cost per time and drug consumption index decreased significantly after the control （P＜0.01）. CONCLUSIONS The drug consumption index is a risk factor for DRG overruns， there is a non-linear relationship and threshold effect between it and DRG overruns. Each hospital can set a reasonable threshold and implement dynamic monitoring and intervention by comprehensively considering the actual drug usage， disease spectrum characteristics， and cost control targets， as well as factors such as medical quality， patient needs， and the payment capacity of medical insurance， which can effectively achieve precise control over drug usage.

ObjectiveTo investigate the effect of Danggui Shaoyaosan on ferroptosis in the rat model of non-alcoholic fatty liver disease （NAFLD） and explore the underlying mechanism based on the nuclear factor E₂-related factor 2 （Nrf2）/solute carrier family 7 member 11 （SLC7A11）/glutathione peroxidase 4 （GPX4） signaling pathway. MethodsThe sixty SD rats were randomly grouped as follows： control， model， Yishanfu （0.144 g·kg^-1）， and low-， medium-， and high-dose （2.44， 4.88， and 9.76 g·kg^-1， respectively） Danggui Shaoyaosan. A high-fat diet was used to establish the rat model of NAFLD. After 12 weeks of modeling， rats were treated with corresponding agents for 4 weeks. Then， the body weight and liver weight were measured， and the liver index was calculated. At the same time， serum and liver samples were collected. The levels or activities of total cholesterol （TC）， triglycerides （TG）， alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， and Fe²⁺ in the serum and TC， TG， free fatty acids （FFA）， malondialdehyde （MDA）， superoxide dismutase （SOD）， glutathione peroxidase （GPX）， and Fe²⁺ in the liver were measured. Hematoxylin-eosin staining and oil red O staining were employed to observe the pathological changes in the liver. Immunofluorescence was used to assess the reactive oxygen species （ROS） content in the liver. Mitochondrial morphology was observed by transmission electron microscopy. The protein levels of Nrf2， SLC7A11， GPX4， transferrin receptor 1 （TFR1）， and divalent metal transporter 1 （DMT1） in the liver were determined by Western blot. ResultsCompared with the control group， the model group showed increases in the body weight， liver weight， liver index， levels or activities of TC， TG， ALT， AST， and Fe²⁺ in the serum， levels of TC， TG， FFA， MDA， Fe²⁺， and ROS in the liver， and protein levels of TFR1 and DMT1 in the liver （P<0.01）， and decreases in the activities of SOD， GPX and the protein levels of Nrf2， SLC7A11， and GPX4 in the liver （P<0.05， P<0.01）. Meanwhile， the liver tissue in the model group presented steatosis， iron deposition， mitochondrial shrinkage， and blurred or swollen mitochondrial cristae. Compared with the model group， all doses of Danggui Shaoyaosan reduced the body weight， liver weight， liver index， levels or activities of TC， TG， ALT， AST， and Fe²⁺ in the serum， levels of TC， TG， FFA， MDA， Fe²⁺， and ROS in the liver， and protein levels of TFR1 and DMT1 in the liver （P<0.01）， while increasing the activities of SOD and GPX and the protein levels of Nrf2， SLC7A11， and GPX4 in the liver （P<0.01）. Furthermore， Danggui Shaoyaosan alleviated steatosis， iron deposition， and mitochondrial damage in the liver. ConclusionDanggui Shaoyaosan may inhibit lipid peroxidation and ferroptosis by activating the Nrf2/SLC7A11/GPX4 signaling pathway to treat NAFLD.

OBJECTIVES: To investigate the impact of incorrect specification of the working correlation structure matrix on estimated sample size in a 2×2 crossover design based on the generalized estimating equation (GEE). METHODS: Based on Monte Carlo simulation, the influence of incorrect specification of the work-related structure matrix on the sample size estimation under different conditions was evaluated after controlling the total sample size n, the proportion of subjects assigned to AB sequence (s=1) θ, the correlation coefficient ρ, and the placebo effect OR. Bias and mean square error (MSE) were used to assess the difference between the sample size estimates and the theoretical values. RESULTS: When the correctly specified working correlation structure matrix is independent, the sample size estimation effect of correctly specifying the working correlation structure matrix is better than that of incorrect specification. But when the correctly specified working correlation structure matrix is equal and the correlation coefficient is closer to 0, with other factors being smaller (n≤50, θ≤0.5, OR=2 in this article), there is a situation where the bias of the sample size estimation value for the correctly specified working correlation structure matrix is greater than the bias for the incorrectly specified working correlation structure matrix. CONCLUSIONS Under most conditions, incorrectly specifying the working correlation structure matrix can cause the estimated sample size to deviate significantly from the theoretical value, but under certain conditions, the impact of incorrectly specifying the working correlation structure matrix can be small on the estimated sample size.
Sample Size ; Monte Carlo Method ; Humans ; Cross-Over Studies ; Computer Simulation ; Research Design ; Bias

As a supplement to randomized controlled trials， observational studies can provide evidence for the effectiveness of traditional Chinese medicine （TCM） treatment measures. They can also study influencing factors of diseases， etiology， and prognosis. However， there is a confounding effect due to the lack of randomization， which seriously affects the causal inference between the study factors and the outcome， resulting in confounding bias. Therefore， identifying and controlling confounding factors are key issues to be addressed in TCM observational studies. According to the causal network and the characteristics of TCM theory， confounding factors can be categorized into measured and unmeasured confounding factors. In addition， attention must be paid to identifying confounding factors and intermediate variables， as well as the interaction between confounding factors and study factors. For methods of controlling confounding factors， measured confounding factors can be controlled by stratification， multifactor analysis， propensity scores， and disease risk scores. Unmeasured and unknown confounding factors can be corrected using instrumental variable methods， difference-in-difference methods， and correction for underlying event rate ratios. Correcting and controlling confounding factors can ensure a balance between groups， and confounding bias can be reduced. In addition， methods such as sensitivity analysis and determination of interactions make the control of confounding factors more comprehensive. Due to the unique characteristics of TCM， observational studies of TCM face unique challenges in identifying and controlling confounding factors， including the ever-changing TCM treatment measures received by patients， the often-overlooked confounding effects in the four diagnostic information of TCM， and the lack of objective criteria for TCM evidence-based diagnosis. Some scholars have already conducted innovative explorations to address these issues， providing a methodological basis for conducting higher-quality TCM observational studies， so as to obtain more rigorous real-world evidence of TCM and gradually develop quality evaluation criteria for OS that are consistent with the characteristics of TCM.

WANG Qishi put forward the theory of "yang deficiency with three lackings， controlled by the spleen" in Lixu Yuanjian （《理虚元鉴》）， which regarded that yang deficiency can lead to consumptive diseases with changes of lacking essence， lacking qi， and lacking fire， so the treatment should start from the spleen to restore the middle yang urgently. This article summarised the experience of treating type 4 cardiorenal syndrome based on the theory of "yang deficiency with three lackings， controlled by the spleen"， and proposed that lacking essence is the beginning of the onset of type 4 cardiorenal syndrome， lacking qi is the gradual development of the disease， and lacking fire is the changes of the disease， and ultimately resulted in the complex situation of kidney and qi deficiency， and edema due to yang deficiency， combined with syndromes variation. In the clinical evidence， in the stage of lacking fire， therapies should warm the middle and strengthen the spleen in order to rescue the middle yang， prescribed with modified Baoyuan Decoction （保元汤） plus Lizhong Decoction （理中汤）； in the stage of lacking qi， prescriptions can add Taoren （Juglans regia）， Tubiechong （Eupolyphaga sinensis）， Fuling （Smilax glabra）， Guizhi （Neolitsea cassia） to activate blood and drain water to transport and restore the center qi； in the stage of lacking essence， prescriptions can add Gouqizi （Lycium barbarum）， Tusizi （Cuscuta chinensis）， Duzhong （Eucommia ulmoides）， Bajitian （Gynochthodes officinalis） to supplement deficiency and resolve masses to consolidate the root and supplement essence.

ObjectiveTo investigate the regulatory effect of Danggui Shaoyaosan on adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/Unc-51-like kinase-1 （ULK1） signaling pathway in the rat model of metabolism-associated fatty liver disease （MAFLD）. MethodSixty SD rats were randomized into control， model， western medicine （polyene phosphatidylcholine capsules，0.144 g·kg^-1）， and low-， medium-， and high-dose （2.44， 4.88， 9.76 g·kg^-1， respectively） Danggui Shaoyaosan groups. After being fed with a high-fat diet for 8 weeks， the rats in each group were administrated with corresponding drugs for 4 weeks. At the end of drug treatment， serum and liver tissue were collected for subsequent determination of related indicators. ResultCompared with the control group， the model group showed increased contents of total cholesterol （TC）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C） and activities of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in the serum， increased contents of TC， TG， and free fatty acids （FFAs） in the liver （P<0.01）， and decreased content of high-density lipoprotein cholesterol （HDL-C） in the serum （P<0.01）. Furthermore， the model group showed down-regulated protein levels of p-AMPK， microtubule-associated protein 1 light chain 3B （LC3B） Ⅱ， Beclin1， and ULK1 （P<0.01） and up-regulated protein levels of p-mTOR and ubiquitin-binding protein p62 in the liver （P<0.01）. The hepatic steatosis was obvious and the NAFLD activity score （NAS） and oil red O staining area increased in the model group， （P<0.05， P<0.01）. Compared with the model group， Danggui Shaoyaosan reduced the contents of TC and TG and the activities of ALT and AST in the serum， lowered the levels of TC， TG， and FFA in the liver， down-regulated the protein levels of p-mTOR and p62 （P<0.01）， elevated the serum HDL-C level， and up-regulated the protein levels of p-AMPK， LCBⅡ， Beclin1， and ULK1 in the liver （P<0.05， P<0.01）. Moreover， it alleviated hepatic steatosis and decreased the NAS and oil red O staining area （P<0.05， P<0.01）. ConclusionDanggui Shaoyaosan has therapeutic effect on MAFLD rats by regulating AMPK/mTOR/ULK1 signaling pathway to enhance autophagy.

Respiratory syncytial virus (RSV) is an enveloped, negative-sense, single-stranded RNA virus of the Orthopneumovirus  genus of the Pneumoviridae family in the order Mononegavirales. RSV can cause acute upper and lower respiratory tract infections, sometimes with extrapulmonary complications. The disease burden of RSV infection is enormous, mainly affecting infants and older adults aged 75 years or above. Currently, treatment options for RSV are largely supportive. Prevention strategies remain a critical focus, with efforts centered on vaccine development and the use of prophylactic monoclonal antibodies. To date, three RSV vaccines have been approved for active immunization among individuals aged 60 years and above. For children who are not eligible for these vaccines, passive immunization is recommended. A newly approved prophylactic monoclonal antibody, Nirsevimab, which offers enhanced neutralizing activity and an extended half-life, provides exceptional protection for high-risk infants and young children. This review provides a comprehensive and detailed exploration of RSV's virology, immunology, pathogenesis, epidemiology, clinical manifestations, treatment options, and prevention strategies.
Humans ; Respiratory Syncytial Virus Infections/prevention & control* ; Respiratory Syncytial Viruses/pathogenicity* ; Respiratory Syncytial Virus, Human/pathogenicity* ; Antiviral Agents/therapeutic use*

Immune imbalance is believed to play a dominant role in the pathogenesis of chronic urticaria.While Th1/Th2 imbalance used to be considered as the main contributing factor of the development of chronic urticaria.Recently,Th17/Treg imbalance is found to be an important immune mechanism leading to the development of chronic urticaria.To be more specific,according to traditional Chinese medicine(TCM)'s comprehensive understanding of the etiology of chronic urticaria.it is generally believed that the pathogenesis of chronic urticaria is due to a lack of innate endowment,a lack of solidity of the body surface,and repeated exposure to six pathogenic factors.Another possible reason lies with dietary disorders that generate heat and wind or chronic illness and weakness and loss of nourishment of qi and blood.Therefore,in terms of the treatment,from the perspective of sthenia syndrome,it is advisable to remove the wind and disperse the pathogenic factors and clear the dampness and heat.From the perspective of asthenia syndrome,it is advisable to nourish the qi and blood and support the righteousness.As for mixed excess and deficiency,both support the healthy atmosphere and dispel the pathogenic factors are important.Regarding the effects of TCM on the balance of Th17/Treg in chronic urticaria and immune diseases,it mainly involved herbal compounding,herbal active ingredients and single herbs.However,the research attention has been drawn to investigating the role of TCM in the treatment of chronic urticaria and various immune diseases based on the research outcomes in modern pharmacological research.This can not only provide scientific evidence for TCM treatment of chronic urticaria,but also bring benefits to more patients with immune diseases.Therefore,the author reviews the recent research progress of TCM on the effects of Th17/Treg immune imbalance in chronic urticaria and other immune diseases by explaining the effects of Th17 and Treg cells in chronic urticaria.

Objective:To explore the relationship and underlying mechanism between exosomes derived from doxorubicin-resistant osteosarcoma cells and MDR1 and miRNAs. Methods:MG63 and U2OS cell lines were selected to construct doxorubicin-resistant strains, and the 50% inhibitory concentration (half maximal inhibitory concentration, IC 50) of drug-resistant and sensitive strains was detected by MTT, and fluorescence staining was performed at intervals of 15 min between 15 and 120 min to detect the change of fluorescence intensity. RT-PCR and Western Blot were used to detect the expression levels of MDR1 P-gp to verify the drug resistance of osteosarcoma cells. Exosomes were identified by particle size analysis and Western Bolt detection. The endocytosis of PKH26-labeled exosomes from doxorubicin-resistant cells was observed, and the proliferation level and migration of exosomes from doxorubicin-resistant cells co-cultured with osteosarcoma cells were detected by MTT assay and cell scratch assay. The differential expression levels of miRNAs in osteosarcoma-sensitive and drug-resistant cells were verified by sequencing and bioinformatics analysis and RT-PCR assay. Tumor growth, serum exosome identification and mRNA expression level of miR-21-5p in tumor-bearing nude mice between normal osteosarcoma cell group and drug-resistant group, drug-resistant+normal exosome group, drug-resistant+drug-resistant+drug-resistant exosome group were observed. MDR1 expression level in tumor tissue was detected by RT-PCR, Western Blot and immunohistochemistry. Results:The IC 50 of two adriamycin resistant strains were 2.21 vs. 11.81 μg/ml and 0.93 vs. 11.81 μg/ml, respectively, and the fluorescence intensity decreased faster than that of normal strains. The relative mRNA expression levels of MDR1 in two cell lines were normal 1.12±0.16, 1.02±0.11 and drug-resistant 2.15±0.10, 2.127±0.12, respectively. The relative protein expression of P-gp was normal 0.92±0.11, 0.73±0.10 and drug-resistant 0.46±0.03, 0.30±0.04, the differences were statistically significant ( P<0.05). Drug-resistant exosomes can enter osteosarcoma cells through endocytosis and concentrate in the cytoplasm when co-cultured with normal strains. Osteosarcoma cells were co-cultured with drug-resistant exosomes at 2, 4, 6, and 8 μg/ml adriamycin, respectively. Compared with normal group, the proliferation level in drug-resistant group was significantly increased. Compared with the normal cell group 35.95±3.92, 6.72±3.55 and the normal exosome group 51.22±5.55, 19.31±1.93, the drug-resistant cell group 54.20±9.32, 19.24±2.88 and drug-resistant exosome group 76.40±5.41, 30.26±4.87, all had significantly higher cell mobility, the difference was statistically significant ( P<0.05). Exosome sequencing and biogenic analysis of 10 highly upregated miRNAs to validate mRNA expression differences between normal and drug-resistant strains by RT-PCR, showing a significant increase in miR-21-5p expression level of drug-resistant strains (5.89±0.26 vs. 0.99±0.06; 1.05±0.07 vs. 8.80±0.93, P<0.05), the difference was statistically significant ( P<0.05). In MG63 and U2OS, the normal cell group and drug-resistant cell group, and the normal exosome group and drug-resistant exosome group were compared, the tumor volume and the terminal tumor weight of nude mice were increased to varying degrees. MRNA relative expression levels of miR-21-5p in serum exosomes of nude mice after drug intervention were 0.86±0.07 and 0.86±0.05 in normal cell group, respectively. The values were 1.13±0.12, 1.14±0.12 in drug-resistant cell group, 0.71±0.05, 0.75±0.03 in normal exosome group, and 0.90±0.07, 0.93±0.04 in drug-resistant exosome group. Compared with normal and drug-resistant strains, the expression levels of normal and drug-resistant exosome groups were increased, with statistical significance ( P<0.05). Conclusion:The exosomes of drug-resistant cells in osteosarcoma could enhance the proliferation level and migration ability of cells through intercellular transfer of MDR1 and miRNAs. The expression of MDR1 and miR-21-5p in drug-resistant cells and tumor-forming nude mouse serum and tumor tissues were up-regulated which suggested that it might be involved in regulating the drug resistance process of osteosarcoma.

Stroke is a cerebrovascular disease that damages brain tissue due to blockage or rupture of blood vessels, and is one of the leading causes of death and disability worldwide. Diagnosing, monitoring stroke and the treatment effects all rely on neuroimaging that detects changes in the brain. Near-infrared spectroscopy (NIRS) is a novel neuroimaging technique which is based on the non-invasively measuring of concentration changes of oxyhemoglobin and reduced hemoglobin. This review summarizes the underlying mechanisms of NIRS and its current clinical application in stroke, as well as the limitations of NIRS in broader clinical applications.