Objective To investigate the changes in eosinophil counts and the activation of microglial cells in the brain tissues of mice at different stages of Angiostrongylus cantonensis infection, and to examine the role of microglia in regulating the progression of angiostrongyliasis and unravel the possible molecular mechanisms. Methods Fifty BALB/c mice were randomly divided into the control group and the 7-d, 14-d, 21-day and 25-d infection groups, of 10 mice in each group. All mice in infection groups were infected with 30 stage III A. cantonensis larvae by gavage, and animals in the control group was given an equal amount of physiological saline. Five mice were collected from each of infection groups on days 7, 14, 21 d and 25 d post-infection, and 5 mice were collected from the control group on the day of oral gavage. The general and focal functional impairment was scored using the Clark scoring method to assess the degree of mouse neurological impairment. Five mice from each of infection groups were sacrificed on days 7, 14, 21 d and 25 d post-infection, and 5 mice from the control group were sacrificed on the day of oral gavage. Mouse brain tissues were sampled, and the pathological changes of brain tissues were dynamically observed using hematoxylin and eosin (HE) staining. Immunofluorescence staining with eosinophilic cationic protein (ECP) and ionized calcium binding adaptor molecule 1 (Iba1) was used to assess the degree of eosinophil infiltration and the counts of microglial cells in mouse brain tissues in each group, and the morphological parameters of microglial cells (skeleton analysis and fractal analysis) were quantified by using Image J software to determine the morphological changes of microglial cells. In addition, the expression of M1 microglia markers Fcγ receptor III (Fcgr3), Fcγ receptor IIb (Fcgr2b) and CD86 antigen (Cd86), M2 microglia markers Arginase 1 (Arg1), macrophage mannose receptor C-type 1 (Mrc1), chitinase-like 3 (Chil3), and phagocytosis genes myeloid cell triggering receptor expressed on myeloid cells 2 (Trem2), CD68 antigen (Cd68), and apolipoprotein E (Apoe) was quantified using real-time quantitative reverse transcription PCR (RT-qPCR) assay in the mouse cerebral cortex of mice post-infection. Results A large number of A. cantonensis larvae were seen on the mouse meninges surface post-infection, and many neuronal nuclei were crumpled and deeply stained, with a large number of bleeding points in the meninges. The median Clark scores of mouse general functional impairment were 0 (interquartile range, 0), 0 (interquartile range, 0.5), 6 (interquartile range, 1.0), 14 (interquartile range, 8.5) points and 20 (interquartile range, 9.0) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.45, P < 0.01), and the median Clark scores of mouse focal functional impairment were 0 (interquartile range, 0), 2 (interquartile range, 2.5), 7 (interquartile range, 3.0), 18 (interquartile range, 5.0) points and 25 (interquartile range, 6.5) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.72, P < 0.01). The mean scores of mice general and focal functional impairment were all higher in the infection groups than in the control group (all P values < 0.05). Immunofluorescence staining showed a significant difference in the eosinophil counts in mouse brain tissues among the five groups (F = 40.05, P < 0.000 1), and the eosinophil counts were significantly higher in mouse brain tissues in the 14-d (3.08 ± 0.78) and 21-d infection groups (5.97 ± 1.37) than in the control group (1.00 ± 0.28) (both P values < 0.05). Semi-quantitative analysis of microglia immunofluorescence showed a significant difference in the counts of microglial cells among the five groups (F = 17.66, P < 0.000 1), and higher Iba1 levels were detected in mouse brain tissues in 14-d (5.75 ± 1.28), 21-d (6.23 ± 1.89) and 25-d infection groups (3.70 ± 1.30) than in the control group (1.00 ± 0.30) (all P values < 0.05). Skeleton and fractal analyses showed that the branch length [(162.04 ± 34.10) μm vs. (395.37 ± 64.11) μm; t = 5.566, P < 0.05] and fractal dimension of microglial cells (1.30 ± 0.01 vs. 1.41 ± 0.03; t = 5.266, P < 0.05) were reduced in mouse brain tissues in the 21-d infection group relative to the control group. In addition, there were significant differences among the 5 groups in terms of M1 and M2 microglia markers Fcgr3 (F = 48.34, P < 0.05), Fcgr2b (F = 55.46, P < 0.05), Cd86 (F = 24.44, P < 0.05), Arg1 (F = 31.18, P < 0.05), Mrc1 (F = 15.42, P < 0.05) and Chil3 (F = 24.41, P < 0.05), as well as phagocytosis markers Trem2 (F = 21.19, P < 0.05), Cd68 (F = 43.95, P < 0.05) and Apoe (F = 7.12, P < 0.05) in mice brain tissues. Conclusions A. cantonensis infections may induce severe pathological injuries in mouse brain tissues that are characterized by massive eosinophil infiltration and persistent activation of microglia cells, thereby resulting in progressive deterioration of neurological functions.

OBJECTIVE: To observe the effect of moxibustion on small intestinal mucosal immune barrier in rats with diarrhea-predominant irritable bowel syndrome (IBS-D) and explore its underlying mechanisms. METHODS: Of 38 newborn rats from 4 healthy SPF pregnant rats, 12 neonatal rats were randomly selected in a normal group. IBS-D model was prepared by the combined measures for the rest rats, including neonatal maternal separation, acetic acid enema and chronic restraint stress. Twenty-four successfully-modeled rats were randomized into a model group and a moxibustion group, 12 rats in each one. In the moxibustion group, suspending moxibustion was delivered at bilateral "Tianshu" (ST25) and "Shangjuxu" (ST37), 20 min each time, once daily and for 7 consecutive days. Separately, before acetic acid enema (aged 35 days), after modeling (aged 45 days) and after intervention (aged 53 days), the body mass, loose stool rate (LSR) and and the minimum volume threshold when abdominal withdrawal reflex (AWR) scored 3 were observed in the rats of each group. After intervention (aged 53 days), using HE and PAS staining, the morphology of duodenum was observed, the length of villus and the depth of crypt were measured, the ratio of the length of villus to the depth of crypt was calculated; and the numbers of mucosal intraepithelial lymphocytes (IELs) and goblet cells were counted. With ELISA adopted, the contents of γ-interferon (IFN-γ), interleukin-4 (IL-4) and secretory immunoglobulin A (sIgA) in duodenal mucosa of rats were detected. The proportion of T cell subsets in duodenal mucosa was detected using flow cytometry. The microvilli and tight junctions of duodenal mucosal epithelial cells were observed by transmission electron microscopy, and the integrity of duodenal mucosa observed by scanning electron microscopy. RESULTS: Compared with the normal group, for the rats in the model group, the body mass, the minimum volume threshold when AWR scored 3, the length of duodenal villus and the the ratio of the length of villus to the depth of crypt, as well as the proportion of CD₈⁺ T subset were all reduced (P<0.01, P<0.05), the counts of goblet cells in duodenal mucosa decreased (P<0.01); LRS, the proportion of CD₄⁺ T subset and CD₄⁺/CD₈⁺, as well as the contents of IFN-γ, IL-4 and sIgA in duodenal mucosa and IFN-γ/IL-4 were all elevated (P<0.01); and the numbers of IELs rose (P<0.01). The morphology of duodenal mucosa was irregular, the villi got shorter, sparse and scattered, with uneven density. The morphology of epithelial cells was destroyed and the tight junctions damaged, with larger spaces. When compared with the model group, in the moxibustion group, the body mass, the minimum volume threshold when AWR scored 3, the length of duodenal villus and the ratio of the length of villus to the depth of crypt, as well as the counts of goblet cells in duodenal mucosa increased (P<0.01); LRS, the proportion of CD₄⁺ T subset, and CD₄⁺/CD₈⁺, as well as the contents of IFN-γ, IL-4 and sIgA in duodenal mucosa and IFN-γ/IL-4 were reduced (P<0.01); and the numbers of IELs was dropped (P<0.01). The morphology of duodenal mucosa was more regular, the villi were grew, got longer and arranged regularly, with even density. The morphology of epithelial cells was slightly destroyed, and the tight junctions partially damaged. CONCLUSION Moxibustion at "Tianshu" (ST25) and "Shangjuxu" (ST37) can reduce visceral hypersensitivity in IBS-D rats and relieve abdominal pain, diarrhea and other symptoms. Its effect mechanism may be related to the repair of small intestinal mucosal immune barrier and the improvement in the immune function in IBS-D.
Animals ; Irritable Bowel Syndrome/immunology* ; Rats ; Moxibustion ; Intestinal Mucosa/immunology* ; Female ; Diarrhea/therapy* ; Intestine, Small/immunology* ; Male ; Humans ; Rats, Sprague-Dawley ; Disease Models, Animal

Objective:In the background of the national promotion of DIP insurance payment reform,using empirical data to evaluate the initial effects of payment reforms in typical traditional Chinese medicine hospitals.Methods:It collected inpatient case data from a Chinese medicine hospital in S city,encompassing a total of 207 500 cases from 2016 to 2022,and the differences between before and after the reform were analyzed using key indicators such as medical costs.Interrupted time series was applied to analyze the initial effects of the reform based on three intervention time points:reform implementation,epidemic development and static management.Logistic regression was applied to explore the main influencing factors,and recommendations for refinement were made based on the results of the key informant interviews.Results:It was found that the percentage of the high hospitalization cost group,high self-pay cost group and high bed-day cost group increased after the reform.There was an overall increasing trend in bed-day costs after the reform.Age,being married,operation times and average grade of operations,and death were the common influences.Conclusion:The initial stage of the DIP insurance payment reform was not effective in regulating medical costs.Demographic changes,strong intervention in epidemics,and the tendency of Chinese medicine hospitals to perform surgeries all tend to lead to an increase in hospitalization costs.At the same time,factors such as the shortage of policy interpretation and the lack of incentives for medical staff have affected the effectiveness of the reform.It was suggested that the DIP insurance payment reform of the Chinese medicine hospitals should be further promoted by strengthening the interpretation of policies and changing the solidification model,establishing positive incentives for medical staff,and adding indicators of traditional Chinese medicine characteristics.This will further fully leverage the advantages of Chinese medicine characteristics and enhance the substance and quality of Chinese medicine services.

To summarize XU Jingshi's clinical experience in treating fever from the perspective of latent pathogen. It is believed that latent pathogen heat accumulation is an important pathogenesis of fever. For latent pathogen in shaoyang and membrane-source， modified Xiaochaihu Decoction （小柴胡汤） plus Qinghao （Artemisiae Annuae Herba） could be used to expel pathogen through membrane and regulate shaoyang； for latent pathogen leading to yin fluids consumption， treatment should clear heat and expel pathogen， and prescribed with Chaihu （Bupleuri Radix）， Huangqin （Scutellaria baicalensis Georgi）， Qinghao （Artemisia carvifolia） combined with Baihu Decoction （白虎汤） or Zhuye Shigao Decoction （竹叶石膏汤） or Qinghao Biejia Powder （青蒿鳖甲散） or Shengmai Decoction （生脉饮） or Siyin Decoction （四阴煎） by syndrome differentiation； for latent pathogen leading to liver wind or reverse transmission to the pericardium， treatment should cool to open pathways， and prescribed with modified Angong Niuhuang Pill （安宫牛黄丸） to clear mind and open the orifices， clear heat and resolve toxins， resolve constraint with aromatics， and expel pathogens to outside； for fever after radiotherapy， chemotherapy or surgery for tumours， prescribed with self-prescribed Fuzheng Anzhong Decoction （扶正安中汤） plus Xiaochaihu Decoction （小柴胡汤） and modified Qinghao Biejia Decoction （青蒿鳖甲汤） to reinforce healthy qi and dispel pathogen， and support the middle jiao.

Objective:In the background of the national promotion of DIP insurance payment reform,using empirical data to evaluate the initial effects of payment reforms in typical traditional Chinese medicine hospitals.Methods:It collected inpatient case data from a Chinese medicine hospital in S city,encompassing a total of 207 500 cases from 2016 to 2022,and the differences between before and after the reform were analyzed using key indicators such as medical costs.Interrupted time series was applied to analyze the initial effects of the reform based on three intervention time points:reform implementation,epidemic development and static management.Logistic regression was applied to explore the main influencing factors,and recommendations for refinement were made based on the results of the key informant interviews.Results:It was found that the percentage of the high hospitalization cost group,high self-pay cost group and high bed-day cost group increased after the reform.There was an overall increasing trend in bed-day costs after the reform.Age,being married,operation times and average grade of operations,and death were the common influences.Conclusion:The initial stage of the DIP insurance payment reform was not effective in regulating medical costs.Demographic changes,strong intervention in epidemics,and the tendency of Chinese medicine hospitals to perform surgeries all tend to lead to an increase in hospitalization costs.At the same time,factors such as the shortage of policy interpretation and the lack of incentives for medical staff have affected the effectiveness of the reform.It was suggested that the DIP insurance payment reform of the Chinese medicine hospitals should be further promoted by strengthening the interpretation of policies and changing the solidification model,establishing positive incentives for medical staff,and adding indicators of traditional Chinese medicine characteristics.This will further fully leverage the advantages of Chinese medicine characteristics and enhance the substance and quality of Chinese medicine services.

Objective:In the background of the national promotion of DIP insurance payment reform,using empirical data to evaluate the initial effects of payment reforms in typical traditional Chinese medicine hospitals.Methods:It collected inpatient case data from a Chinese medicine hospital in S city,encompassing a total of 207 500 cases from 2016 to 2022,and the differences between before and after the reform were analyzed using key indicators such as medical costs.Interrupted time series was applied to analyze the initial effects of the reform based on three intervention time points:reform implementation,epidemic development and static management.Logistic regression was applied to explore the main influencing factors,and recommendations for refinement were made based on the results of the key informant interviews.Results:It was found that the percentage of the high hospitalization cost group,high self-pay cost group and high bed-day cost group increased after the reform.There was an overall increasing trend in bed-day costs after the reform.Age,being married,operation times and average grade of operations,and death were the common influences.Conclusion:The initial stage of the DIP insurance payment reform was not effective in regulating medical costs.Demographic changes,strong intervention in epidemics,and the tendency of Chinese medicine hospitals to perform surgeries all tend to lead to an increase in hospitalization costs.At the same time,factors such as the shortage of policy interpretation and the lack of incentives for medical staff have affected the effectiveness of the reform.It was suggested that the DIP insurance payment reform of the Chinese medicine hospitals should be further promoted by strengthening the interpretation of policies and changing the solidification model,establishing positive incentives for medical staff,and adding indicators of traditional Chinese medicine characteristics.This will further fully leverage the advantages of Chinese medicine characteristics and enhance the substance and quality of Chinese medicine services.

Objective:In the background of the national promotion of DIP insurance payment reform,using empirical data to evaluate the initial effects of payment reforms in typical traditional Chinese medicine hospitals.Methods:It collected inpatient case data from a Chinese medicine hospital in S city,encompassing a total of 207 500 cases from 2016 to 2022,and the differences between before and after the reform were analyzed using key indicators such as medical costs.Interrupted time series was applied to analyze the initial effects of the reform based on three intervention time points:reform implementation,epidemic development and static management.Logistic regression was applied to explore the main influencing factors,and recommendations for refinement were made based on the results of the key informant interviews.Results:It was found that the percentage of the high hospitalization cost group,high self-pay cost group and high bed-day cost group increased after the reform.There was an overall increasing trend in bed-day costs after the reform.Age,being married,operation times and average grade of operations,and death were the common influences.Conclusion:The initial stage of the DIP insurance payment reform was not effective in regulating medical costs.Demographic changes,strong intervention in epidemics,and the tendency of Chinese medicine hospitals to perform surgeries all tend to lead to an increase in hospitalization costs.At the same time,factors such as the shortage of policy interpretation and the lack of incentives for medical staff have affected the effectiveness of the reform.It was suggested that the DIP insurance payment reform of the Chinese medicine hospitals should be further promoted by strengthening the interpretation of policies and changing the solidification model,establishing positive incentives for medical staff,and adding indicators of traditional Chinese medicine characteristics.This will further fully leverage the advantages of Chinese medicine characteristics and enhance the substance and quality of Chinese medicine services.

Objective:In the background of the national promotion of DIP insurance payment reform,using empirical data to evaluate the initial effects of payment reforms in typical traditional Chinese medicine hospitals.Methods:It collected inpatient case data from a Chinese medicine hospital in S city,encompassing a total of 207 500 cases from 2016 to 2022,and the differences between before and after the reform were analyzed using key indicators such as medical costs.Interrupted time series was applied to analyze the initial effects of the reform based on three intervention time points:reform implementation,epidemic development and static management.Logistic regression was applied to explore the main influencing factors,and recommendations for refinement were made based on the results of the key informant interviews.Results:It was found that the percentage of the high hospitalization cost group,high self-pay cost group and high bed-day cost group increased after the reform.There was an overall increasing trend in bed-day costs after the reform.Age,being married,operation times and average grade of operations,and death were the common influences.Conclusion:The initial stage of the DIP insurance payment reform was not effective in regulating medical costs.Demographic changes,strong intervention in epidemics,and the tendency of Chinese medicine hospitals to perform surgeries all tend to lead to an increase in hospitalization costs.At the same time,factors such as the shortage of policy interpretation and the lack of incentives for medical staff have affected the effectiveness of the reform.It was suggested that the DIP insurance payment reform of the Chinese medicine hospitals should be further promoted by strengthening the interpretation of policies and changing the solidification model,establishing positive incentives for medical staff,and adding indicators of traditional Chinese medicine characteristics.This will further fully leverage the advantages of Chinese medicine characteristics and enhance the substance and quality of Chinese medicine services.

Objective:In the background of the national promotion of DIP insurance payment reform,using empirical data to evaluate the initial effects of payment reforms in typical traditional Chinese medicine hospitals.Methods:It collected inpatient case data from a Chinese medicine hospital in S city,encompassing a total of 207 500 cases from 2016 to 2022,and the differences between before and after the reform were analyzed using key indicators such as medical costs.Interrupted time series was applied to analyze the initial effects of the reform based on three intervention time points:reform implementation,epidemic development and static management.Logistic regression was applied to explore the main influencing factors,and recommendations for refinement were made based on the results of the key informant interviews.Results:It was found that the percentage of the high hospitalization cost group,high self-pay cost group and high bed-day cost group increased after the reform.There was an overall increasing trend in bed-day costs after the reform.Age,being married,operation times and average grade of operations,and death were the common influences.Conclusion:The initial stage of the DIP insurance payment reform was not effective in regulating medical costs.Demographic changes,strong intervention in epidemics,and the tendency of Chinese medicine hospitals to perform surgeries all tend to lead to an increase in hospitalization costs.At the same time,factors such as the shortage of policy interpretation and the lack of incentives for medical staff have affected the effectiveness of the reform.It was suggested that the DIP insurance payment reform of the Chinese medicine hospitals should be further promoted by strengthening the interpretation of policies and changing the solidification model,establishing positive incentives for medical staff,and adding indicators of traditional Chinese medicine characteristics.This will further fully leverage the advantages of Chinese medicine characteristics and enhance the substance and quality of Chinese medicine services.

Objective:In the background of the national promotion of DIP insurance payment reform,using empirical data to evaluate the initial effects of payment reforms in typical traditional Chinese medicine hospitals.Methods:It collected inpatient case data from a Chinese medicine hospital in S city,encompassing a total of 207 500 cases from 2016 to 2022,and the differences between before and after the reform were analyzed using key indicators such as medical costs.Interrupted time series was applied to analyze the initial effects of the reform based on three intervention time points:reform implementation,epidemic development and static management.Logistic regression was applied to explore the main influencing factors,and recommendations for refinement were made based on the results of the key informant interviews.Results:It was found that the percentage of the high hospitalization cost group,high self-pay cost group and high bed-day cost group increased after the reform.There was an overall increasing trend in bed-day costs after the reform.Age,being married,operation times and average grade of operations,and death were the common influences.Conclusion:The initial stage of the DIP insurance payment reform was not effective in regulating medical costs.Demographic changes,strong intervention in epidemics,and the tendency of Chinese medicine hospitals to perform surgeries all tend to lead to an increase in hospitalization costs.At the same time,factors such as the shortage of policy interpretation and the lack of incentives for medical staff have affected the effectiveness of the reform.It was suggested that the DIP insurance payment reform of the Chinese medicine hospitals should be further promoted by strengthening the interpretation of policies and changing the solidification model,establishing positive incentives for medical staff,and adding indicators of traditional Chinese medicine characteristics.This will further fully leverage the advantages of Chinese medicine characteristics and enhance the substance and quality of Chinese medicine services.