ObjectiveTo investigate whether Huanglian Jiedutang can inhibit neutrophil infiltration in the brains of middle cerebral artery occlusion （MCAO） mice by regulating the expression of neutrophil-related chemokines in exosomes， thereby achieving therapeutic effects. MethodsA total of 130 male specific pathogen-free （SPF） C57BL/6J mice were randomly divided into four groups： Sham-operated group， MCAO model group， Huanglian Jiedutang group （6 g·kg^-1）， and Ginaton group （21.6 mg·kg^-1）， with 10 mice in the Ginaton group and 40 mice in each of the remaining three groups. Mice in the Huanglian Jiedutang group and the Ginaton group were administered the corresponding drugs by oral gavage once daily at a volume of 0.15 mL·（10 g）^-1 for 7 consecutive days， while the sham-operated and model groups received an equal volume of saline via the same route. After 7 days， MCAO surgery was performed. The distal and proximal ends of the right common carotid artery （CCA） were ligated， a small incision was made between the two ligatures， and a silicone rubber-coated monofilament with a rounded tip was inserted into the lumen to occlude the CCA. The filament was left in place for 1 h to establish a focal cerebral ischemia model. At 24 h after modeling， mice were evaluated. Neurological function was assessed using the Longa score. Cerebral infarct volume was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Cerebral blood flow was observed by laser speckle imaging. Hematoxylin and eosin （HE） staining and Nissl staining were used to observe pathological changes in brain tissues. Exosomes were isolated from mouse plasma and brain tissues by ultracentrifugation and molecular size exclusion and identified by electron microscopy， particle size analysis， and protein blotting. Long-chain RNA libraries of exosomes were constructed and sequenced. Real-time quantitative reverse transcription polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of inflammatory factors and neutrophil-related chemokines in exosomes from plasma and brain tissues of each group. Enzyme-linked immunosorbent assay （ELISA） was used to detect the protein expression of inflammatory factors and neutrophil-related chemokines in exosomes from brain tissues of each group. Immunohistochemistry was used to detect the expression of the neutrophil-specific protein myeloperoxidase （MPO） in the brains of mice in each group. ResultsCompared with the sham-operated group， the model group showed decreased neurological function scores （P<0.01）， obvious cerebral infarction （P<0.01）， reduced cerebral blood flow （P<0.01）， neuronal necrosis in the brain， and decreased numbers of Nissl bodies （P<0.01）. The mRNA expression levels of IL-1β， MPO， CXCL1， CXCL2， CXCL3， CXCL10， CCL2， and CCL3 in exosomes from plasma and brain tissues were significantly increased （P<0.05， P<0.01）. The protein expression levels of IL-1β， MPO， CXCL2， and CXCL10 in exosomes from brain tissues were increased （P<0.05， P<0.01）， and MPO-positive rates and mean optical density values in brain tissues were elevated （P<0.01）. Compared with the model group， the Huanglian Jiedutang group and the Ginaton group showed increased neurological function scores （P<0.05）， reduced cerebral infarct volume （P<0.01）， restored cerebral blood flow （P<0.01）， reduced necrotic cells in the brain， and increased numbers of Nissl bodies （P<0.01）. In the Huanglian Jiedutang group， the mRNA expression levels of IL-1β， MPO， CXCL1， CXCL2， CXCL3， CXCL10， CCL2， and CCL3 in exosomes from plasma and brain tissues were decreased （P<0.05， P<0.01）. The protein expression levels of IL-1β， MPO， CXCL2， and CXCL10 in exosomes from brain tissues were reduced （P<0.05， P<0.01）， and MPO-positive rates and mean optical density values in brain tissues were decreased （P<0.01）. ConclusionHuanglian Jiedutang can effectively regulate the expression of neutrophil-related chemokines in exosomes from plasma and brain tissues of MCAO mice， thereby reducing neutrophil infiltration in the brain and achieving therapeutic effects.

ObjectiveTo investigate whether Huanglian Jiedutang can inhibit neutrophil infiltration in the brains of middle cerebral artery occlusion （MCAO） mice by regulating the expression of neutrophil-related chemokines in exosomes， thereby achieving therapeutic effects. MethodsA total of 130 male specific pathogen-free （SPF） C57BL/6J mice were randomly divided into four groups： Sham-operated group， MCAO model group， Huanglian Jiedutang group （6 g·kg^-1）， and Ginaton group （21.6 mg·kg^-1）， with 10 mice in the Ginaton group and 40 mice in each of the remaining three groups. Mice in the Huanglian Jiedutang group and the Ginaton group were administered the corresponding drugs by oral gavage once daily at a volume of 0.15 mL·（10 g）^-1 for 7 consecutive days， while the sham-operated and model groups received an equal volume of saline via the same route. After 7 days， MCAO surgery was performed. The distal and proximal ends of the right common carotid artery （CCA） were ligated， a small incision was made between the two ligatures， and a silicone rubber-coated monofilament with a rounded tip was inserted into the lumen to occlude the CCA. The filament was left in place for 1 h to establish a focal cerebral ischemia model. At 24 h after modeling， mice were evaluated. Neurological function was assessed using the Longa score. Cerebral infarct volume was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Cerebral blood flow was observed by laser speckle imaging. Hematoxylin and eosin （HE） staining and Nissl staining were used to observe pathological changes in brain tissues. Exosomes were isolated from mouse plasma and brain tissues by ultracentrifugation and molecular size exclusion and identified by electron microscopy， particle size analysis， and protein blotting. Long-chain RNA libraries of exosomes were constructed and sequenced. Real-time quantitative reverse transcription polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of inflammatory factors and neutrophil-related chemokines in exosomes from plasma and brain tissues of each group. Enzyme-linked immunosorbent assay （ELISA） was used to detect the protein expression of inflammatory factors and neutrophil-related chemokines in exosomes from brain tissues of each group. Immunohistochemistry was used to detect the expression of the neutrophil-specific protein myeloperoxidase （MPO） in the brains of mice in each group. ResultsCompared with the sham-operated group， the model group showed decreased neurological function scores （P<0.01）， obvious cerebral infarction （P<0.01）， reduced cerebral blood flow （P<0.01）， neuronal necrosis in the brain， and decreased numbers of Nissl bodies （P<0.01）. The mRNA expression levels of IL-1β， MPO， CXCL1， CXCL2， CXCL3， CXCL10， CCL2， and CCL3 in exosomes from plasma and brain tissues were significantly increased （P<0.05， P<0.01）. The protein expression levels of IL-1β， MPO， CXCL2， and CXCL10 in exosomes from brain tissues were increased （P<0.05， P<0.01）， and MPO-positive rates and mean optical density values in brain tissues were elevated （P<0.01）. Compared with the model group， the Huanglian Jiedutang group and the Ginaton group showed increased neurological function scores （P<0.05）， reduced cerebral infarct volume （P<0.01）， restored cerebral blood flow （P<0.01）， reduced necrotic cells in the brain， and increased numbers of Nissl bodies （P<0.01）. In the Huanglian Jiedutang group， the mRNA expression levels of IL-1β， MPO， CXCL1， CXCL2， CXCL3， CXCL10， CCL2， and CCL3 in exosomes from plasma and brain tissues were decreased （P<0.05， P<0.01）. The protein expression levels of IL-1β， MPO， CXCL2， and CXCL10 in exosomes from brain tissues were reduced （P<0.05， P<0.01）， and MPO-positive rates and mean optical density values in brain tissues were decreased （P<0.01）. ConclusionHuanglian Jiedutang can effectively regulate the expression of neutrophil-related chemokines in exosomes from plasma and brain tissues of MCAO mice， thereby reducing neutrophil infiltration in the brain and achieving therapeutic effects.

Objective:To analyze trends in urticaria incidence among children aged 0 - 14 years in China from 1990 to 2021, to explore its changing patterns in different age, period, and cohort groups, and to investigate the impact of age and air pollutants on the incidence trends.Methods:Data were obtained from the Global Burden of Disease Database (GBD2021) , including the number of urticaria cases, crude incidence rates, and age-standardized incidence rates among children aged 0 - 14 years of different genders in China from 1990 to 2021. The Joinpoint regression model was used to calculate the annual percentage change (APC) and average annual percentage change (AAPC) to assess temporal trends in incidence rates. An age-period-cohort model was applied to assess the effects of age, period, and cohort on urticaria incidence. Data on the annual emissions of 4 air pollutants (SO 2, CO, PM 2.5, and PM 10) in China from 1990 to 2021 were obtained from the Emissions Database for Global Atmospheric Research (EDGAR) , and a multivariable meta-regression model was used to explore the relationship between air pollutants and urticaria incidence. Results:From 1990 to 2021, the age-standardized incidence rate of urticaria among children aged 0 - 14 years in China demonstrated a slight overall downward trend (AAPC = -0.03%, P < 0.01) . The incidence rate was generally higher in female children than in male children, and the decline in incidence rates was greater in female children than in male children (female AAPC = -0.02%, male AAPC = -0.01%, both P < 0.01) . The age-period-cohort model indicated that the risk of urticaria decreased with advancing age: with the age group of 0 - 4 years as the reference ( RR = 1.000) , the risk of urticaria significantly decreased in the age group of 5 - 9 years ( RR = 0.790, 95% CI: 0.789 - 0.791) and further declined in the age group of 10 - 14 years ( RR = 0.711, 95% CI: 0.710 - 0.711) ; the period effect analysis showed that the risk of urticaria gradually decreased after the baseline period of 1992 - 1996 ( RR = 1.000) , and dropped to 0.995 (95% CI: 0.994 - 0.997) in the period of 2017 - 2021; in the cohort effect analysis of the overall population aged 0 - 14 years, with the 1988 - 1992 birth cohort as the base cohort, an earlier birth cohort 1978 - 1982 exhibited the highest risk of urticaria ( RR = 1.006, 95% CI: 1.004 - 1.009) , while the 2013 - 2017 cohort showed the lowest risk ( RR = 0.996, 95% CI: 0.994 - 0.997) . The multivariable meta-regression analysis indicated a significant association between PM 2.5 exposure and urticaria incidence ( β = 0.319, 95% CI: 0.022 - 0.616, P = 0.033) , although this association was not statistically significant in different age groups. Conclusions:From 1990 to 2021, children aged 0 - 4 years in China were the highest-risk group for urticaria; the decline in the incidence rate of urticaria was more pronounced in female children than in male children, and earlier birth cohorts exhibited higher risks of urticaria. Exposure to PM 2.5 appeared to be associated with the incidence of urticaria.

Objective:To analyze the impact of DRG payment method reform on hospitalization costs and structure in traditional Chinese medicine hospitals.Methods:Using the cost data of hospitalized patients from 2017 to 2022 for public traditional Chinese medicine hospitals in Qingyang and Tianshui,descriptive statistics and two groups interrupted time-series models were implemented to compare the changes before and after DRG payment method reform.Results:DRG payment method reform had no effective impact on the secondary traditional Chinese medicine hospital's average hospitalization costs,average Chinese medicine costs,and average western medicine costs(P＞0.05),but the average proportion of western medicine costs showed an obvious downward trend after the reform(β9=-0.19,P＜0.01).DRG payment method reform resulted in tertiary traditional Chinese medicine hospitals turning to obvious decreasing trends in average hospitalization costs,average Chinese medicine costs,and average western medicine costs(P＜0.05),while the pre-reform decreasing trends in the average proportion Chinese medicine costs share of both secondary and tertiary traditional Chinese medicine hospitals were curbed in the post-reform period.Conclusion:DRG payment method reform had effectively controlled hospitalization costs,Chinese medicine costs,and western medicine costs of tertiary traditional Chinese medicine hospitals,while the cost control of secondary traditional Chinese medicine hospitals has been ineffective,probably contributing to the enhancement of the use of Chinese medicine and the average proportion of Chinese medicine costs.DRG payment method reform for Chinese medicine should strengthen the supervision of costs,actively utilize the advantages of the characteristics of Chinese medicine,and promote the differentiated reform of traditional Chinese medicine hospitals.

Objective:It aims to examine the effects of the employee medical insurance outpatient mutual assistance on medical insurance costs for patients with diabetes mellitus,offering insights for optimizing outpatient insurance policies and chronic disease management strategies.Methods:Outpatient cost settlement data of urban employees with diabetes mellitus in Lanzhou from 2022 to 2023 was collected.Univariate analysis and interrupted time-series models were used to compare relevant medical insurance cost indicators before and after the reform.Results:The inpatient data of 765 730 diabetes mellitus patients were included in the study,and male patients account for 62.67％.After the reform,average per-visit pooling fund expenditure,average per-visit individual payment expenditures,average per-visit personal account expenditure,average per-visit eligible expense amount,average per-visit total fund payment,and the average per-visit proportion of basic medical pooling payments were decreased(P＜0.05).Moreover,average per-visit pooling fund expenditure,average per-visit individual account expenditure,average per-visit cash payment,average per-visit eligible expense amount,average per-visit total fund payment,and the average per-visit proportion of basic medical pooling payments showed a notable declining trend post-reform(P＜0.05).In contrast,the per-visit fully out-of-pocket expenditure exhibited no significant change before and after the reform(P＞0.05).Conclusion:The reform of the employee medical insurance outpatient mutual assistance system has alleviated the economic burden of disease for diabetic patients and improved the efficiency of medical insurance fund utilization,but it reduced the proportion of basic medical pooling payments.It is recommended to continuously refine the outpatient medical insurance payment system,strengthen supervision of medical expenses and service quality,and balance patient benefits with fund pressure to enhance chronic disease outpatient benefits.

Objective:To analyze trends in urticaria incidence among children aged 0 - 14 years in China from 1990 to 2021, to explore its changing patterns in different age, period, and cohort groups, and to investigate the impact of age and air pollutants on the incidence trends.Methods:Data were obtained from the Global Burden of Disease Database (GBD2021) , including the number of urticaria cases, crude incidence rates, and age-standardized incidence rates among children aged 0 - 14 years of different genders in China from 1990 to 2021. The Joinpoint regression model was used to calculate the annual percentage change (APC) and average annual percentage change (AAPC) to assess temporal trends in incidence rates. An age-period-cohort model was applied to assess the effects of age, period, and cohort on urticaria incidence. Data on the annual emissions of 4 air pollutants (SO 2, CO, PM 2.5, and PM 10) in China from 1990 to 2021 were obtained from the Emissions Database for Global Atmospheric Research (EDGAR) , and a multivariable meta-regression model was used to explore the relationship between air pollutants and urticaria incidence. Results:From 1990 to 2021, the age-standardized incidence rate of urticaria among children aged 0 - 14 years in China demonstrated a slight overall downward trend (AAPC = -0.03%, P < 0.01) . The incidence rate was generally higher in female children than in male children, and the decline in incidence rates was greater in female children than in male children (female AAPC = -0.02%, male AAPC = -0.01%, both P < 0.01) . The age-period-cohort model indicated that the risk of urticaria decreased with advancing age: with the age group of 0 - 4 years as the reference ( RR = 1.000) , the risk of urticaria significantly decreased in the age group of 5 - 9 years ( RR = 0.790, 95% CI: 0.789 - 0.791) and further declined in the age group of 10 - 14 years ( RR = 0.711, 95% CI: 0.710 - 0.711) ; the period effect analysis showed that the risk of urticaria gradually decreased after the baseline period of 1992 - 1996 ( RR = 1.000) , and dropped to 0.995 (95% CI: 0.994 - 0.997) in the period of 2017 - 2021; in the cohort effect analysis of the overall population aged 0 - 14 years, with the 1988 - 1992 birth cohort as the base cohort, an earlier birth cohort 1978 - 1982 exhibited the highest risk of urticaria ( RR = 1.006, 95% CI: 1.004 - 1.009) , while the 2013 - 2017 cohort showed the lowest risk ( RR = 0.996, 95% CI: 0.994 - 0.997) . The multivariable meta-regression analysis indicated a significant association between PM 2.5 exposure and urticaria incidence ( β = 0.319, 95% CI: 0.022 - 0.616, P = 0.033) , although this association was not statistically significant in different age groups. Conclusions:From 1990 to 2021, children aged 0 - 4 years in China were the highest-risk group for urticaria; the decline in the incidence rate of urticaria was more pronounced in female children than in male children, and earlier birth cohorts exhibited higher risks of urticaria. Exposure to PM 2.5 appeared to be associated with the incidence of urticaria.

Objective:It aims to examine the effects of the employee medical insurance outpatient mutual assistance on medical insurance costs for patients with diabetes mellitus,offering insights for optimizing outpatient insurance policies and chronic disease management strategies.Methods:Outpatient cost settlement data of urban employees with diabetes mellitus in Lanzhou from 2022 to 2023 was collected.Univariate analysis and interrupted time-series models were used to compare relevant medical insurance cost indicators before and after the reform.Results:The inpatient data of 765 730 diabetes mellitus patients were included in the study,and male patients account for 62.67％.After the reform,average per-visit pooling fund expenditure,average per-visit individual payment expenditures,average per-visit personal account expenditure,average per-visit eligible expense amount,average per-visit total fund payment,and the average per-visit proportion of basic medical pooling payments were decreased(P＜0.05).Moreover,average per-visit pooling fund expenditure,average per-visit individual account expenditure,average per-visit cash payment,average per-visit eligible expense amount,average per-visit total fund payment,and the average per-visit proportion of basic medical pooling payments showed a notable declining trend post-reform(P＜0.05).In contrast,the per-visit fully out-of-pocket expenditure exhibited no significant change before and after the reform(P＞0.05).Conclusion:The reform of the employee medical insurance outpatient mutual assistance system has alleviated the economic burden of disease for diabetic patients and improved the efficiency of medical insurance fund utilization,but it reduced the proportion of basic medical pooling payments.It is recommended to continuously refine the outpatient medical insurance payment system,strengthen supervision of medical expenses and service quality,and balance patient benefits with fund pressure to enhance chronic disease outpatient benefits.

Objective:To analyze the impact of DRG payment method reform on hospitalization costs and structure in traditional Chinese medicine hospitals.Methods:Using the cost data of hospitalized patients from 2017 to 2022 for public traditional Chinese medicine hospitals in Qingyang and Tianshui,descriptive statistics and two groups interrupted time-series models were implemented to compare the changes before and after DRG payment method reform.Results:DRG payment method reform had no effective impact on the secondary traditional Chinese medicine hospital's average hospitalization costs,average Chinese medicine costs,and average western medicine costs(P＞0.05),but the average proportion of western medicine costs showed an obvious downward trend after the reform(β9=-0.19,P＜0.01).DRG payment method reform resulted in tertiary traditional Chinese medicine hospitals turning to obvious decreasing trends in average hospitalization costs,average Chinese medicine costs,and average western medicine costs(P＜0.05),while the pre-reform decreasing trends in the average proportion Chinese medicine costs share of both secondary and tertiary traditional Chinese medicine hospitals were curbed in the post-reform period.Conclusion:DRG payment method reform had effectively controlled hospitalization costs,Chinese medicine costs,and western medicine costs of tertiary traditional Chinese medicine hospitals,while the cost control of secondary traditional Chinese medicine hospitals has been ineffective,probably contributing to the enhancement of the use of Chinese medicine and the average proportion of Chinese medicine costs.DRG payment method reform for Chinese medicine should strengthen the supervision of costs,actively utilize the advantages of the characteristics of Chinese medicine,and promote the differentiated reform of traditional Chinese medicine hospitals.

Objective To investigate the effect of low salt diet on vascular remodeling of rat induced by high fructose(HF).Methods Wistar male rats weighed 180-200 g were fed for 8 weeks and randomly divided into 6 groups:(1) control group was given the normal fodder and distilled water;(2) high fructose group(HF) was given normal fodder (0.5 ％ NaCl,w/w) and fructose water(10 ％,w/v);(3) high-salt group (HNa) was given high salt fodder (7 ％ NaCl,w/w) and distilled water;(4) high fructose combined with high salt diet group(HFNa) was simultaneously given high salt fodder and 10 ％ fructose water;(5)high fructose combined low salt group(HFLNa) was simultaneously given low salt fodder and 10％ fructose water;(6) high fructose combined with spirotaclone group(HFE) was given 10％ fructose water for 4 weeks and then added with spirotaelone(50 mg · kg-1 · d-1 by tube feeding) for continuous 4 weeks.The changes of arterial blood pressure,vascular wall histological evaluation and expression of α-SMA and fibronectin in vascular wall were detected in each group.Results (1) Compared with the blood pressure[(111.03 ±9.17) mm Hg] in the control group,the blood pressure in the HF and HNa groups were (133.94± 5.86) mm Hg and (128.09±7.56) mm Hg respectively,which were significantly increased(P＜0.05);(2) HF mainly caused the hyperplasia of vascular wall middle layer smooth muscle.The a-SMA expression results in the HF group was (0.006 3 ±0.000 21),which in the control group was (0.004 6 ± 0.000 31),the difference was statistically significant(P＜0.05),moreover which promoted the elastic fibers increase;while HNa mainly stimulated the elastic fibers to thicken and extracellular matrix deposition,the fibronectin expression was 0.002 6 ± 0.000 2 in the HNa group and (0.004 7±0.000 2)in the HF group,compared with(0.001 3±0.000 1)in the normal group,which were significantly increased(P＜0.001);(3) the blood pressure was (106.04±9.59) mm Hg in the HFLNa group,(103.99±7.12) mm Hg in the HFE group,compared with(133.94±5.86) mm Hg in the HF group,showing that the blood pressure in the HFLNa group and HFE group was significantly decreased compared with the HF group (P＜0.05);moreover the vascular remodeling in the HFLNa group(0.006 8±0.000 2) and HFE group (0.004 2±0.000 4) was improved,and compared with the HF group(0.006 3±0.000 2),α-SMA expression was significantly decreased (P＜0.05).Conclusion Low salt diet can effectively improve vascular remodeling induced by HEF.

Objective To study the factor affecting the efficiency of establishment of parthenogenetic stem cells (PESCs)in mice. Methods PESC lines were derived from parthenogenetically activated blastocyst inner cell mass of different mice strains and cultured in different systems.Results The efficiency of establishing the pESC showed no significant difference in hybrid and inbred lines.But the efficiency was increased in culture systems added with ERK inhibitor or knockout serum replacement(KSR).Conclusion The efficiency of establishment of pESC was not directly related with the genetic background of mice but it was closely related to the culture systems.