ObjectiveTo explore the potential mechanism underlying electroacupuncture therapy with the midnight-noon ebb-low method with hour-prescription for chronic obstructive pulmonary disease （COPD） with lung-qi deficiency syndrome from the perspective of brain and muscle basic helix-loop-helix ARNT-like 1 （BMAL1） as well as neutrophil extracellular traps （NETs）. MethodsThe experiment was conducted in two phases. For the therapeutic effectiveness observation， 24 rats were randomly allocated into control group 1， model group 1， midnight-noon electro-acupuncture group 1 and conventional electro-acupuncture group， with 6 rats per group. Rat models of COPD with lung-qi deficiency syndrome were established via cigarette smoke exposure combined with intratracheal instillation of lipopolysaccharide （LPS）. After successful modelling， rats in midnight-noon electro-acupuncture group 1 received electro-acupuncture at acupoints "Taiyuan （LU 9）"， "Feishu （BL 13）" and "Zusanli （ST 36）" during 05：00—07：00， 30 minutes per treatment once every other day for 14 consecutive days （7 sessions in total）. Rats in the conventional electro-acupuncture group received identical electroacupuncture manipulation at random daytime hours （08：00—18：00）. Pulmonary function parameters including forced expiratory volume in 0.3 second （FEV_0.3）， forced vital capacity （FVC） and FEV_0.3/FVC ratio were detected. Pulmonary histopathological changes were observed via hematoxylin-eosin （HE） staining. Plasma levels of pro-inflammatory factors interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α）， as well as pulmonary reactive oxygen species （ROS） content， BMAL1 protein expression and the levels of neutrophil extracellular traps （NETs） biomarkers such as myeloperoxidase （MPO）， neutrophil elastase （NE）， and citrullinated histone H3 （CitH3） in lung tissue were determined. For the mechanistic verification phase， 36 rats were divided into control group 2， model group 2， midnight-noon electro-acupuncture group 2， overexpression empty group， overexpression BMAL1 group and overexpression with midnight-noon electro-acupuncture group， with 6 rats in each group. At the 5th week of model construction， rats in the overexpression BMAL1 group and overexpression with midnight-noon electro-acupuncture group received intratracheal instillation of 100 μl adenoviral suspension carrying overexpressed Bmal1 gene. Rats in the overexpression empty group were injected with an equal titer and equal volume of blank adenovirus without the Bmal1 coding sequence into the lung at the identical time point. Midnight-noon electro-acupuncture group 2 and overexpression with midnight-noon electro-acupuncture group were consistent with the aforementioned protocol. Finally， pulmonary levels of MPO， NE and CitH3 were measured in all groups. ResultsCompared with the control group 1， the model group 1 exhibited decreased FEV_0.3， FVC and FEV_0.3/FVC ratio， elevated plasma IL-1β， TNF-α and pulmonary ROS levels， downregulated lung BMAL1 protein expression， and increased contents of MPO， NE and CitH3 （all P＜0.05）； typical NETs-like structures with sparse granular substances attached to fibrous networks were observed under scanning electron microscopy； HE staining revealed damaged alveolar architecture accompanied by inflammatory cell infiltration. Compared with model group 1， both the midnight-noon electro-acupuncture group 1 and conventional electro-acupuncture group achieved obvious improvements in all above indicators， with superior therapeutic effects in midnight-noon electro-acupuncture group 1 （P＜0.05）. Consistently， midnight-noon electro-acupuncture group 1 showed more prominent alleviation of NETs-like structure formation and lung pathological injury compared to the conventional electro-acupuncture group. Mechanistic validation results demonstrated that compared with model group 2， the average optical density of MPO as well as the protein levels of MPO， NE and CitH3 were markedly reduced in midnight-noon electro-acupuncture group 2， overexpression BMAL1 group and overexpression with midnight-noon electro-acupuncture group （P＜0.05）； furthermore， these parameters were significantly lower in the overexpression with midnight-noon electro-acupuncture group than in midnight-noon electro-acupuncture group 2 （P＜0.05）. ConclusionElectro-acupuncture based on the midnight-noon ebb-low method with hour-prescription may alleviate pulmonary inflammation in COPD by upregulating the expression of clock protein BMAL1 and inhibiting excessive accumulation of NETs in lung tissue.

Objective: To analyze the epidemiological characteristics of hand, foot and mouth disease (HFMD) among children aged 0-9 years in Guangzhou, so as to provide reference basis for further exploring scientifically effective prevention and control measures. Methods: Reported cases of HFMD in children aged 0-9 years in Guangzhou City from 2013-2025 were selected from China disease prevention and control information system, and the incidence in different years was analyzed descriptively; the trends of the composition ratio and incidence in various age groups were analyzed by using the Joinpoint regression model, spatial autocorrelation analyses were performed at the district level, and the seasonal autoregressive integrated moving average model (SARIMA) was applied to predict annual incidence rate for 2026. Results: From 2013 to 2025, the average annual reported incidence rate of HFMD among children aged 0-9 years in Guangzhou was 2 513.98/100 000, showing a fluctuating trend. Peaks of incidence occurred from May to July and from September to October. The male to female ratio was 1.49∶1. The average annual incidence rate was 4 743.99/100 000 in the 0-1 years age group, 3 298.19/100 000 in the 2-5 years age group, and 544.22/ 100 000 in the 6-9 years age group. Joinpoint regression analysis showed a decreasing trend in the proportion of the 0-1 years age group (APC= -6.02 %, t =-3.77) and an increasing trend in the proportion of the 6-9 years age group (APC=16.39%, t =5.67) (both P < 0.05 ). No significant global spatial clustering was observed at the district level; however, local scattered clusters were identified in certain areas. Using the SARIMA(2,0,0)(2,0,0) 12 model, the predicted annual incidence rate of HFMD among children aged 0-9 years in Guangzhou in 2026 was 2 680.56/100 000. Conclusions From 2013 to 2025, the annual incidence rate of HFMD among children aged 0-9 years in Guangzhou exhibited a fluctuating trend; the highest incidence rate is observed in the 0-1 years age group, while the proportion of cases in the 6-9 years age group increase year by year;and local spatial clusters of abnormal distribution are identified. It is essential to enhance risk monitoring and health education in key populations and areas to mitigate the risk of infection in children.

Objective: To explore the role of N6-methyladenosine(m6A) modification of RNA and Methyltransferase like Protein 14 methyltransferase(METTL14) in the pathogenesis of early-onset preeclampsia(ePE). Methods: Placental tissues of 15 pregnant women with early-onset preeclampsia and 15 normal pregnant women were collected. The level of m6A was determined by colorimetry, and the expression of METTL14 was determined by RT-qPCR, Western blot and immunohistochemistry(IHC) experiments. By transfecting siRNA and plasmid, METTL14 levels of trophoblast cells were knocked down and overexpressed, and cell phenotype experiments were carried out in vitro. The effects of METTL14 on the proliferation, migration and invasion of trophoblast cells were investigated by CCK-8, scratch assay, Transwell assay and invasion assay. Results: The level of m6A in placental tissue of ePE was lower than that of normal pregnancy. METTL14 was mainly expressed in the nuclei of trophoblast cells. Compared with normal pregnancy, the expression of METTL14 in placental tissue of ePE decreased, and thelevel of METTL14 mRNA was positively correlated with the level of m6A in placental tissue. The results of the CCK-8 experiment showed that compared with the control group, knockdown of METTL14 expression in trophoblast cells significantly reduced the cell proliferation rate, while the proliferation ability of trophoblast cells with overexpressed METTL14 was enhanced. The results of the scratch test showed that compared with the control group, the relative healing rate of scratches was significantly reduced after METTL14 knockdown, while it increased after the overexpression of METTL14. The results of the Transwell assay and invasion assay showed that compared with the control group, after knockdown of METTL14, the number of trophoblast cells passing through the chamber was significantly reduced, while the number of trophoblast cells with overexpressed METTL14 passing through the chamber increased. Conclusion The total RNA m6A modification level in placental tissue of ePE is lower than that in the normal pregnancy group. The down-regulation of methyltransferase METTL14 is involved in the regulation of the total RNA m6A modification level. The overexpression of METTL14 can enhance the proliferation, migration and invasion abilities of trophoblast cells. It provides a new perspective for exploring the pathogenesis of early-onset preeclampsia.

Objective To observe the effects of Huoxue Tongjiang Decoction on the expressions of gelsolin(GSN),prosaposin(PSAP)and nuclear factor(NF)-κB in reflux esophagitis(RE)model rats;To explore its mechanism of protecting esophageal mucosa.Methods Totally 75 Wistar rats were randomly divided into control group,model group,Western medicine group,TCM low-and high-dosage groups,with 15 rats in each group.The control group was given celiac switch operation,the other groups were given modified partial cardiac myotomy and external pyloric partial ligation to prepare RE rat model.Seven days after operation,each group was given normal saline,Western medicine or corresponding dosage of Huoxue Tongjiang Decoction for gavage,twice a day for 14 days.HE staining was used to observe the morphology of esophageal mucosa and pathological score was evaluated,immunohistochemical staining was used to detect the positive expressions of GSN and PSAP in esophageal mucosa,and Western blot was used to detect the protein expressions of NF-κBp65 and p-NF-κBp65 in esophageal tissue,ELISA was used to detect the contents of serum tumor necrosis factor(TNF)-α and interleukin(IL)-6.Results Compared with the control group,the esophageal mucosa of the model group was obviously damaged,the pathological score increased,the expression of GSN in esophageal mucosa decreased,the expressions of PSAP,NF-κBp65 and p-NF-κBp65 protein increased,and the contents of TNF-α and IL-6 in serum increased,with statistical significance(P<0.01).Compared with the model group,the esophageal mucosal injury improved obviously in the Western medicine group and TCM low-and high-dosage groups,the pathological score decreased,the expression of GSN in esophageal mucosa increased,the expressions of PSAP,NF-κBp65 and p-NF-κBp65 protein decreased,and the contents of TNF-α and IL-6 in serum decreased,with statistical significance(P<0.05,P<0.01).Conclusion Huoxue Tongjiang Decoction can effectively improve the immune inflammation of esophageal mucosa in RE model rats,its mechanism may be related to regulating GSN,PSAP and NF-κB pathway and reducing the release of inflammatory factors TNF-α and IL-6.

Objective To explore the effect of moxibustion on bone metabolism and the mechanism of bone protection,the effect of moxibustion,a traditional Chinese treatment in patients with rheumatoid arthritis(RA),especially the levels of two biomarkers,interleukin 35(IL-35)and β-catenin in the serum of rheumatoid arthritis patients were investigated.Methods A total of 72 RA patients were randomly assigned equally to the treatment group(36 cases)and 36 patients each of the control group(36 cases).The control group was intervened with standardized Western medicine treatment.For the treatment group,moxibustion intervened on Zusanli,Shenshu,and Ashi acupoints three times a week for 5-week treatment,with the remaining treatments being the same as the control group.The clinical symptom-related scores,routine clinical test indicators,and serum IL-35,β-catenin,BMP-2,IL-1β contents of RA patients before and after treatment were compared between the two groups.Results After treatment,VAS score,DAS28 score,morning stiffness score and BMP-2,β-catenin and IL-1β in serum in 2 groups significantly decreased(P<0.01),while IL-35 content in treatment group significantly increased(P<0.01).VAS score,DAS28 score,morning stiffness score and BMP-2,β-catenin and IL-1β in serum showed the obvious advantage of the treatment group over the control group(P<0.05),but the two groups(P>0.05)have no significant discrepancy in serum IL-35 content.For the treatment group,the improvement values of morning stiffness score and joint swelling number were positively correlated with the improvement values of BMP-2 and β-catenin in serum(P<0.01,P<0.05,r>0).The improvement of serum IL-35 was negatively correlated with the improvement of DAS28 score in the treatment group(P<0.05,r<0).Conclusion On the basis of conventional western medicine treatment,moxibustion may inhibit the levels of IL-35 and IL-1β in serum of RA patients,regulate the Wnt/β-catenin signaling pathway,regulate the levels of β-catenin and BMP-2,reduce the inflammatory response of RA,regulate bone metabolism,and achieve the purpose of bone protection.Thus,bone and joint teratogenesis or disability in RA patients was prevented and delayed.

Objective To explore the effect of moxibustion on bone metabolism and the mechanism of bone protection,the effect of moxibustion,a traditional Chinese treatment in patients with rheumatoid arthritis(RA),especially the levels of two biomarkers,interleukin 35(IL-35)and β-catenin in the serum of rheumatoid arthritis patients were investigated.Methods A total of 72 RA patients were randomly assigned equally to the treatment group(36 cases)and 36 patients each of the control group(36 cases).The control group was intervened with standardized Western medicine treatment.For the treatment group,moxibustion intervened on Zusanli,Shenshu,and Ashi acupoints three times a week for 5-week treatment,with the remaining treatments being the same as the control group.The clinical symptom-related scores,routine clinical test indicators,and serum IL-35,β-catenin,BMP-2,IL-1β contents of RA patients before and after treatment were compared between the two groups.Results After treatment,VAS score,DAS28 score,morning stiffness score and BMP-2,β-catenin and IL-1β in serum in 2 groups significantly decreased(P<0.01),while IL-35 content in treatment group significantly increased(P<0.01).VAS score,DAS28 score,morning stiffness score and BMP-2,β-catenin and IL-1β in serum showed the obvious advantage of the treatment group over the control group(P<0.05),but the two groups(P>0.05)have no significant discrepancy in serum IL-35 content.For the treatment group,the improvement values of morning stiffness score and joint swelling number were positively correlated with the improvement values of BMP-2 and β-catenin in serum(P<0.01,P<0.05,r>0).The improvement of serum IL-35 was negatively correlated with the improvement of DAS28 score in the treatment group(P<0.05,r<0).Conclusion On the basis of conventional western medicine treatment,moxibustion may inhibit the levels of IL-35 and IL-1β in serum of RA patients,regulate the Wnt/β-catenin signaling pathway,regulate the levels of β-catenin and BMP-2,reduce the inflammatory response of RA,regulate bone metabolism,and achieve the purpose of bone protection.Thus,bone and joint teratogenesis or disability in RA patients was prevented and delayed.

Objective To observe the effects of Huoxue Tongjiang Decoction on the expressions of gelsolin(GSN),prosaposin(PSAP)and nuclear factor(NF)-κB in reflux esophagitis(RE)model rats;To explore its mechanism of protecting esophageal mucosa.Methods Totally 75 Wistar rats were randomly divided into control group,model group,Western medicine group,TCM low-and high-dosage groups,with 15 rats in each group.The control group was given celiac switch operation,the other groups were given modified partial cardiac myotomy and external pyloric partial ligation to prepare RE rat model.Seven days after operation,each group was given normal saline,Western medicine or corresponding dosage of Huoxue Tongjiang Decoction for gavage,twice a day for 14 days.HE staining was used to observe the morphology of esophageal mucosa and pathological score was evaluated,immunohistochemical staining was used to detect the positive expressions of GSN and PSAP in esophageal mucosa,and Western blot was used to detect the protein expressions of NF-κBp65 and p-NF-κBp65 in esophageal tissue,ELISA was used to detect the contents of serum tumor necrosis factor(TNF)-α and interleukin(IL)-6.Results Compared with the control group,the esophageal mucosa of the model group was obviously damaged,the pathological score increased,the expression of GSN in esophageal mucosa decreased,the expressions of PSAP,NF-κBp65 and p-NF-κBp65 protein increased,and the contents of TNF-α and IL-6 in serum increased,with statistical significance(P<0.01).Compared with the model group,the esophageal mucosal injury improved obviously in the Western medicine group and TCM low-and high-dosage groups,the pathological score decreased,the expression of GSN in esophageal mucosa increased,the expressions of PSAP,NF-κBp65 and p-NF-κBp65 protein decreased,and the contents of TNF-α and IL-6 in serum decreased,with statistical significance(P<0.05,P<0.01).Conclusion Huoxue Tongjiang Decoction can effectively improve the immune inflammation of esophageal mucosa in RE model rats,its mechanism may be related to regulating GSN,PSAP and NF-κB pathway and reducing the release of inflammatory factors TNF-α and IL-6.

Objective:To analyze epidemiological characteristics and spatiotemporal clustering of hepatitis A in China from 2005 to 2023.Methods:The incidence data of hepatitis A in China during 2005-2023 were collected from the China Disease Control and Prevention Information System .Results:From 2005 to 2023, a total of 605 509 cases of hepatitis A were reported in China. The average annual reported incidence rate was 2.32/100 000, the incidence was 2.85/100 000 in men and 1.81/100 000 in women, and the incidence was 3.25/100 000 in age group 0-14 years, 2.10/100 000 in age group 15-64 years and 2.49/100 000 in age group ≥65 years , respectively. The case count in farmers was highest (40.57%, 245 639/605 509). The proportion of the cases in jobless or the unemployed increased most obviously (347.32%), and the proportion of the cases in students decreased most significantly (90.27%). The average annual reported incidence rate in the western China was highest (4.45/100 000), followed by that in northeastern China (2.02/100 000), central China (1.89/100 000) and eastern China (1.16/100 000). From 2020 to 2023, the incidence of hepatitis A showed no spatial clustering. From 2005 to 2019, the obvious hot spots and high-high clustering areas mainly distributed in provinces, such as, Sichuan, Qinghai, Xinjiang, Xizang and Gansu. The low-low clustering areas were mainly distributed in Beijing, Tianjin, Hebei, Shandong, Jiangsu, Shanghai, Zhejiang, Fujian, Henan, Anhui, Jiangxi, Jilin and Liaoning. An independent high-low clustering was found in Shanxi during 2014-2019. A total of 5 high incidence clustering areas were detected through spatiotemporal scanning analysis.Conclusions:The incidence rate of hepatitis A in China declined significantly from 2005 to 2023. The reported incidence rate in the elderly showed no obvious decrease, and the reported cases accounted for the highest proportion in the whole population. Before 2020, the reported incidence rate of hepatitis A showed high-high clustering in western China, the spatiotemporal clustering disappeared from 2020 to 2023, but the reported incidence rate of hepatitis A in western China was still high. It is suggested to pay attention to the prevention and control of hepatitis A in populations at high risk and areas with high incidence of hepatitis A.

Objective:To analyze the nationwide epidemiological characteristics and trend of hand, foot and mouth disease (HFMD) fatal cases from 2008 to 2022 and provide evidence for the prevention and control of HFMD.Methods:The information on HFMD fatal cases during 2008 to 2022 was collected from the National Notifiable Disease Surveillance Reporting System of China. Data of the epidemiological characteristics was analyzed by R 4.2.2 software and the changing trends for the case fatality rates, mortality rates and their age-adjusted rates were analyzed by Joinpoint 4.9.10 software.Results:From 2008 to 2022, a total of 3 704 fatal HFMD cases were reported in China. The fatal cases were primarily observed in children aged <3 years (83.42%, 3 090/3 704). The male and female gender ratio was 1.82 ∶1 (2 389 ∶1 315). Regarding the age-adjusted case fatality rates over time, there was a rapid increase from 2008 to 2010 [annual percentage change (APC) =41.97%, P<0.05]. From 2010 to 2016, a steady decline was observed (APC=-28.57%, P<0.05), and the decline accelerated (APC=-39.66%, P<0.05) from 2016 to 2022. Since 2020, less than 10 fatal cases were reported annually nationwide. Among the 2 566 laboratory-confirmed deaths from 2008 to 2022, Enterovirus A71 (EV71) was the predominant pathogen (91.62%,2 351/2 566). There have been noticeable changes in the pathogen composition since 2017, decreasing in EV71 and increasing in the proportion of fatalities caused by Coxsackievirus A16 (CV-A16) and other enteroviruses. Conclusions:From 2008 to 2022, the HFMD case fatality rates and mortality rates continuously declined, peaked in 2010. Since 2017, the decline of HFMD case fatality rates has been noticeably accelerated. Along with the decrease in the proportion of EV71 in HFMD fatal cases, the proportion of other enteroviruses appeared increasing. It is essential to continuously monitor the etiological spectrum of the fatal cases.