ObjectiveTo explore the interaction and competitive binding of Homo sapiens long intergenic non-protein-coding RNA 1134 （LINC01134） to CCCTC-binding factor CTCF， affecting the transcription of cyclin-dependent kinase inhibitor （p21） and influencing the proliferation of A549 cells， in order to investigate the possible mechanism of Astragali Radix and Hedyotis diffusa （A-H） in inhibiting A549 proliferation by regulating this axis. MethodsRNA-binding protein immunoprecipitation （RIP） assays were conducted to examine the interaction between LINC01134 and CTCF， and chromatin immunoprecipitation （ChIP） assays were used to study the effect of LINC01134 overexpression on the interaction between CTCF and p21. Stable A549 cell lines （oe-NC and oe-LINC01134） were established using lentiviral transfection， and each group was treated with 10% A-H drug-containing serum. Real-time PCR and Western blot analyses were performed to detect the effects of A-H on the expression of LINC01134， CTCF， and p21 in A549 cells. Cell counting kit-8 （CCK-8） and colony formation assays were used to assess the effects of A-H on A549 cell proliferation via LINC01134. Flow cytometry was employed to evaluate the effects of A-H on the A549 cell cycle through LINC01134， and Western blot was used to detect changes in cell cycle proteins. ResultsCompared with the IgG group， the oe-CTCF group showed a significantly increased abundance of LINC01134 aggregates （P0.01）. Compared with the oe-Vector group， p21 abundance in CTCF complexes was significantly reduced in the oe-LINC01134 group （P0.01）. Compared with the 10% blank + oe-LINC01134 group， the 10% A-H + oe-LINC01134 group reversed the expression of LINC01134 and p21 （P0.05）， but had no significant regulatory effect on CTCF. Compared with the 10% blank + oe-LINC01134 group， the 10% A-H + oe-LINC01134 group reversed cell viability at 72 h （P0.05）， inhibited malignant proliferation （P0.05）， and reversed the proportions of cells in the G₀/G₁ and S phases （P0.01）. Furthermore， compared with the 10% blank + oe-LINC01134 group， the 10% A-H + oe-LINC01134 group reversed the expression of Cyclin D₁， CDK4， Cyclin E， CDK2， phosphorylated retinoblastoma protein （p-Rb）， and E2F transcription factor 3 （E2F3） （P0.01）. ConclusionA-H regulates the LINC01134-CTCF-p21 axis to block the G₁/S phase transition of A549 cell cycle， accelerate cellular senescence， and inhibit malignant proliferation.

ObjectiveTo explore the interaction and competitive binding of Homo sapiens long intergenic non-protein-coding RNA 1134 （LINC01134） to CCCTC-binding factor CTCF， affecting the transcription of cyclin-dependent kinase inhibitor （p21） and influencing the proliferation of A549 cells， in order to investigate the possible mechanism of Astragali Radix and Hedyotis diffusa （A-H） in inhibiting A549 proliferation by regulating this axis. MethodsRNA-binding protein immunoprecipitation （RIP） assays were conducted to examine the interaction between LINC01134 and CTCF， and chromatin immunoprecipitation （ChIP） assays were used to study the effect of LINC01134 overexpression on the interaction between CTCF and p21. Stable A549 cell lines （oe-NC and oe-LINC01134） were established using lentiviral transfection， and each group was treated with 10% A-H drug-containing serum. Real-time PCR and Western blot analyses were performed to detect the effects of A-H on the expression of LINC01134， CTCF， and p21 in A549 cells. Cell counting kit-8 （CCK-8） and colony formation assays were used to assess the effects of A-H on A549 cell proliferation via LINC01134. Flow cytometry was employed to evaluate the effects of A-H on the A549 cell cycle through LINC01134， and Western blot was used to detect changes in cell cycle proteins. ResultsCompared with the IgG group， the oe-CTCF group showed a significantly increased abundance of LINC01134 aggregates （P0.01）. Compared with the oe-Vector group， p21 abundance in CTCF complexes was significantly reduced in the oe-LINC01134 group （P0.01）. Compared with the 10% blank + oe-LINC01134 group， the 10% A-H + oe-LINC01134 group reversed the expression of LINC01134 and p21 （P0.05）， but had no significant regulatory effect on CTCF. Compared with the 10% blank + oe-LINC01134 group， the 10% A-H + oe-LINC01134 group reversed cell viability at 72 h （P0.05）， inhibited malignant proliferation （P0.05）， and reversed the proportions of cells in the G₀/G₁ and S phases （P0.01）. Furthermore， compared with the 10% blank + oe-LINC01134 group， the 10% A-H + oe-LINC01134 group reversed the expression of Cyclin D₁， CDK4， Cyclin E， CDK2， phosphorylated retinoblastoma protein （p-Rb）， and E2F transcription factor 3 （E2F3） （P0.01）. ConclusionA-H regulates the LINC01134-CTCF-p21 axis to block the G₁/S phase transition of A549 cell cycle， accelerate cellular senescence， and inhibit malignant proliferation.

ObjectiveThis paper aims to observe the syndrome improvement and negative antigen conversion rate of Qingxuan Daozhi formula in the treatment of influenza in children （heat accumulation in the lung and stomach syndrome）. MethodsThrough a multi-center randomized controlled methodology design，confirmed influenza cases were collected from October 2022 to April 2023 in the pediatrics department of eight hospitals，such as Dongfang Hospital of Beijing University of Chinese Medicine. A total of 180 children with influenza and heat accumulation in the lung and stomach syndrome conforming to the standard were recruited through the clinic. The sick children meeting the inclusion criteria were randomly divided into groups by a block-randomized method. The children in the experimental group were treated with Qingxuan Daozhi formula for five days，and those in the control group were treated with Oseltamivir Phosphate Granules for five days. The primary efficacy indicator was the negative conversion rate of influenza antigen detection. Secondary efficacy indicators were the Canadian acute respiratory illness and flu scale （CARIFS） and the incidence of complications，severe cases， and critical cases. Follow-up observation was conducted on the day of enrollment，48 hours after medication，72 hours after medication， and （6+1） d after medication. ResultsOne hundred and eighty participants were randomly assigned to the experimental group （90 cases） or the control group （90 cases）. All participants were followed up during the study. Comparison of influenza antigen detection results in the primary efficacy indicators showed that the average time of negative influenza antigen conversion in the experimental group was （5.29±1.25） d，and that in the control group was （5.40±1.68） d，without a statistically significant difference. After five days of intervention，52 cases in the experimental group and 51 cases in the control group converted to negative，without a statistically significant difference. CARIFS score results in the secondary efficacy indicators showed that during 72 hours after intervention，there were statistically significant differences between the experimental group and the control group in three dimensions， including headache，muscle soreness， and the need for extra care （P<0.05）. On the （6+1） days after the intervention，the differences in both the experimental group and the control group were statistically significant in 10 dimensions， including sore throat，bad sleep，uncomfortable feeling，poor spirit and fatigue，crying more than usual，the need for extra care，symptom，function，influence on parents，and total score （P<0.05）. The comparison results within the group in the dimensional scores of symptom， function， and influence on parents，as well as the CARIFS total score showed that with the delay of follow-up time，scores of both groups decreased significantly，with a statistically significant difference （P<0.01）. Inter-group comparison results showed that the mean score of the experimental group was higher than that of the control group at the time of enrollment. With the progress of intervention，the score of the experimental group was significantly decreased compared with that of the control group. At the end of follow-up，the mean score of the experimental group was lower than that of the control group，with no statistically significant difference. In terms of the incidence of complications，severe cases， and critical cases， there were no complications，severe cases， and critical cases in the two groups，without a statistically significant difference. ConclusionThe symptom improvement effect and negative antigen conversion rate of Qingxuan Daozhi formula in the treatment of influenza in children （heat accumulation in the lung and stomach syndrome） are not inferior to Oseltamivir Phosphate granules， and children's acceptance is better. It can be more widely used in clinical treatment of influenza in children （heat accumulation in the lung and stomach syndrome）.

ObjectiveTo investigate the effects of excessive activation of the Specific protein 1 （Sp1）-MicroRNA-582-3p （miR-582-3p）-cyclin-dependent kinase inhibitor （p27） axis on the cell cycle and proliferation of A549 lung adenocarcinoma cells at the cellular level， thereby investigating the possible mechanisms by which Astragali Radix and Hedyotis diffusa（A-H） regulate the axis to inhibit A549 cells proliferation. MethodsLentiviral plasmid transfection technology was used to obtain four stable A549 transgenic cell lines： shRNA-Sp1+LV-miR-582-3p mimic， shRNA-Sp1+LV-mNC， shRNA-NC+LV-miR-582-3p mimic， and shRNA-NC+LV-mNC. Real-time PCR was used to detect the effect of Sp1 on miR-582-3p expression in A549 lung adenocarcinoma cells. CCK-8 and colony formation assay were used to detect the effect of Sp1 on cell proliferation through miR-582-3p， while flow cytometry was used to detect the effect of Sp1 on cell cycle via miR-582-3p. Western blot was used to detect the effect of Sp1 on cyclin in A549 cells through miR-582-3p. CCK-8 was employed to detect the effects of different concentrations （5%， 10%， 20%， 40%， 80%） of A-H on A549 cell viability， and the optimal concentration was selected for subsequent mechanism exploration. Stable transgenic strains oe-Sp1 and control oe-Vector were constructed using the above-mentioned transfection techniques. Real-time PCR was then used to investigate the effect of A-H on miR-582-3p expression in A549 cells， while Western blot was employed to detect the effect of A-H on Sp1 and p27 expression in A549 lung adenocarcinoma cells. ResultsCompared with the shRNA-NC+LV-mNC group， the expression of miR-582-3p was significantly inhibited in the shRNA-Sp1+LV-mNC group （P<0.01）. Compared with the shRNA-Sp1+LV-mNC group， the shRNA-Sp1+LV-miR-582-3p mimic group significantly reversed miR-582-3p expression （P<0.01）， indicating that Sp1 has a significant regulatory effect on miR-582-3p. Compared with the shRNA-Sp1+LV-mNC group， the shRNA-Sp1+LV-miR-582-3p mimic group significantly reversed cell proliferation （P<0.01）， suggesting that miR-582-3p can partially reverse the proliferative effect of Sp1 on A549 lung adenocarcinoma. Compared with the shRNA-Sp1+LV-mNC group， the shRNA-Sp1+LV-miR-582-3p mimic group showed a significant decrease in G₀/G₁ proportions， while the G₂/M and S phase proportions increased significantly （P<0.01）， indicating that miR-582-3p can partially reverse the effect of Sp1 on the A549 cell cycle. Compared with the shRNA-Sp1+LV-mNC group， the shRNA-Sp1+LV-miR-582-3p mimic group significantly reversed the expression of various proteins with the decrease of p27 and the increase of Cyclin D₁， CDK4， Cyclin E， CDK2， p-Rb， and E2F3 （P<0.01）， indicating that miR-582-3p can partially reverse the effect of Sp1 on the cell cycle of A549 lung adenocarcinoma. Compared with the blank group， the serum containing A-H significantly inhibited A549 cell viability in a concentration-dependent manner. Therefore， 10% A-H was selected for intervention of subsequent mechanism experiments. Compared with A-H+oe-Vector， A-H+oe-Sp1 significantly reversed the downregulation of Sp1， miR-582-3p， and the upregulation of p27 （P<0.01）. This suggests that Sp1 can partially reverse the effects of A-H on miR-582-3p and p27 levels in A549 lung adenocarcinoma cells. ConclusionOveractivation of the Sp1-miR-582-3p-p27 axis significantly promotes the proliferation of A549 lung adenocarcinoma cells， and the potential mechanism of the inhibitory effect of A-H on the proliferation of A549 cells may be related to its inhibition on the overactivation of the Sp1-miR-582-3p-p27 axis.

Children， as a special group， frequently experience of off-label drug use worldwide. Common reasons for off-label drug use in children include the lack of data on pediatric patients during the clinical trial stage of drug development， delayed updates to drug instructions， and the non-standard professional behavior of some doctors. Off-label drug use in children is a double-edged sword. It could save lives and provide a way to explore additional functions of drugs， while it may also lead to the phenomenon of hyper-indication abuse， increasing the risk of adverse drug events. Regulating off-label drug use in children can safeguard the best treatment rights and interests of children. It is recommended to encourage pharmaceutical enterprises to conduct research and development of pediatric new drugs， simplify the approval process for drug instructions amendments， accumulate evidence-based medical evidence for off-label drug use in children， standardize the process of off-label drug use in children in medical institutions， continuously improve the standardized diagnosis and treatment capabilities of pediatricians， and actively cooperate with the families of pediatric patients in diagnosis and treatment， so as to comprehensively safeguard the rights and interests of both doctors and patients.

ObjectiveTo evaluate the efficacy and safety of Xiaoji Hufei Formula in protecting children with close contact exposure to influenza， and to provide reference and evidence-based support for better clinical prevention and treatment of influenza in children. MethodsA multicenter， prospective， non-randomized， parallel， controlled trial was conducted from October 2021 to May 2022 in five hospitals， including Dongfang Hospital of Beijing University of Chinese Medicine. Confirmed influenza cases and influenza-like illness （ILI） cases were collected， and eligible children with close contact exposure to these cases were recruited in the outpatient clinics. According to whether the enrolled close contacts were willing to take Xiaoji Hufei formula for influenza prevention， they were assigned to the observation group （108 cases） or the control group （108 cases）. Follow-up visits were conducted on days 7 and 14 after enrollment. The primary outcomes were the incidence of ILI and the rate of laboratory-confirmed influenza. Secondary outcomes included traditional Chinese medicine （TCM） symptom score scale for influenza， influenza-related emergency （outpatient） visit rate， influenza hospitalization rate， and time to onset after exposure to influenza cases. ResultsA total of 216 participants were enrolled， with 108 in the observation group and 108 in the control group. Primary outcomes： （1） Incidence of ILI： The incidence was 12.0% （13/108） in the observation group and 23.1% （25/108） in the control group， with the observation group showing a significantly lower incidence （χ²=4.6， P<0.05）. （2） Influenza confirmation rate： 3.7% （4/108） in the observation group and 4.6% （5/108） in the control group， with no statistically significant difference. Secondary outcomes： （1） TCM symptom score scale： after onset， nasal congestion and runny nose scores differed significantly between the two groups （P<0.05）， while other symptoms such as fever， sore throat， and cough showed no significant differences. （2） Influenza-related emergency （outpatient） visit rate： 84.6% （11 cases） in the observation group and 96.0% （24 cases） in the control group， with no significant difference. （3） Time to onset after exposure： The median onset time after exposure to index patients was 7 days in the observation group and 4 days in the control group， with a statistically significant difference （P<0.05）. ConclusionIn previously healthy children exposed to infectious influenza cases under unprotected conditions， Xiaoji Hufei formula prophylaxis significantly reduced the incidence of ILI. Xiaoji Hufei Formula can be recommended as a specific preventive prescription for influenza in children.

Objective To explore the clinical significance of miR-423-5p in ischemic stroke and its effects on SH-SY5Y cell apoptosis along with the underlying mechanisms.Methods Plasma samples were collected from three stroke-prone patients who subsequently developed acute ischemic stroke(AIS)within 24 hours.High-throughput sequencing was used to identify miR-423-5p as a key target.Venous blood samples were collected from 46 AIS pa-tients within 24 hours of onset and from 46 matched control cases.RT-qPCR was performed to measure the plasma expression of miR-423-5p in both groups.The diagnostic value of miR-423-5p for AIS was evaluated using receiver operating characteristic(ROC)curve.After overexpression and knockdown of miR-423-5p in SH-SY5Y cells,flow cytometry was performed to detect apoptosis in each group to analyze the potential relationship between miR-423-5p and SH-SY5Y cell apoptosis.Western blot was used to measure changes in the expression level of the ap-optotic protein cleaved caspase-3 and the anti-apoptotic proteins Bcl-2 and Bcl-xL.Results The expression of miR-423-5p in 3 AIS patients showed significant difference before and after onset.Plasma miR-423-5p expression was significantly elevated in AIS patients versus controls(P＜0.001).ROC analysis indicated its strong diagnostic potential for AIS.Overexpression of miR-423-5p increased the apoptosis rate and cleaved caspase-3 ex-pression of SH-SY5Y cells after OGD/R,and decreased the expression of Bcl-2 and Bcl-xL,while knocking down miR-423-5p had the opposite effect.Conclusions The expression of miR-423-5p is upregulated in the plasma of patients with ischemic stroke compared to healthy controls.Furthermore,miR-423-5p promotes OGD/R-induced apoptosis in SH-SY5Y cells by increasing the level of cleaved caspase-3 and suppressing the expression of Bcl-2 and Bcl-xL.

ObjectiveTo evaluate the efficacy and safety of Xiaoji Hufei Formula in protecting children with close contact exposure to influenza， and to provide reference and evidence-based support for better clinical prevention and treatment of influenza in children. MethodsA multicenter， prospective， non-randomized， parallel， controlled trial was conducted from October 2021 to May 2022 in five hospitals， including Dongfang Hospital of Beijing University of Chinese Medicine. Confirmed influenza cases and influenza-like illness （ILI） cases were collected， and eligible children with close contact exposure to these cases were recruited in the outpatient clinics. According to whether the enrolled close contacts were willing to take Xiaoji Hufei formula for influenza prevention， they were assigned to the observation group （108 cases） or the control group （108 cases）. Follow-up visits were conducted on days 7 and 14 after enrollment. The primary outcomes were the incidence of ILI and the rate of laboratory-confirmed influenza. Secondary outcomes included traditional Chinese medicine （TCM） symptom score scale for influenza， influenza-related emergency （outpatient） visit rate， influenza hospitalization rate， and time to onset after exposure to influenza cases. ResultsA total of 216 participants were enrolled， with 108 in the observation group and 108 in the control group. Primary outcomes： （1） Incidence of ILI： The incidence was 12.0% （13/108） in the observation group and 23.1% （25/108） in the control group， with the observation group showing a significantly lower incidence （χ²=4.6， P<0.05）. （2） Influenza confirmation rate： 3.7% （4/108） in the observation group and 4.6% （5/108） in the control group， with no statistically significant difference. Secondary outcomes： （1） TCM symptom score scale： after onset， nasal congestion and runny nose scores differed significantly between the two groups （P<0.05）， while other symptoms such as fever， sore throat， and cough showed no significant differences. （2） Influenza-related emergency （outpatient） visit rate： 84.6% （11 cases） in the observation group and 96.0% （24 cases） in the control group， with no significant difference. （3） Time to onset after exposure： The median onset time after exposure to index patients was 7 days in the observation group and 4 days in the control group， with a statistically significant difference （P<0.05）. ConclusionIn previously healthy children exposed to infectious influenza cases under unprotected conditions， Xiaoji Hufei formula prophylaxis significantly reduced the incidence of ILI. Xiaoji Hufei Formula can be recommended as a specific preventive prescription for influenza in children.

Objective:To compare the dosimetric parameters under different incremental modes between intensity-modulated radiation therapy(IMRT)and volume rotation intensity-modulated therapy(VMAT)for the target area of large volume brain metastases(BMs),and to explore the better way of treating BMs based on hypofractionated stereotactic radiotherapy(HSRT)of linear accelerator.Methods:A total of 30 BMs patients who underwent IMRT at The 971th Hospital of Navy of the CPLA from 2020 to 2023 were selected.In the treatment planning system(TPS),three types of IMRT plans and VMAT plans were designed,which included uniformity plan(Planuniformity)of target area dose,uniform increased plan(Planuniform increased-dose)and incremental plan(Planincremental)within target area.In the inside of the target area,the target area of high dose(GTVh)was set,and Planuniform increased-dose and Planincremental were designed to aim at GTVh.The differences of the doses of three types of treatment plans included Planuniformity,Planuniform increased-dose and Planincremental,which were respectively designed by using IMRT and VMAT,were analyzed.The mean dose(Dmean)of the target area,the 50%and 2%exposed doses(D50%and D2%)of the target area were observed and compared.The conformity index(CI),homogeneity index(HI),gradient index(GI),and the volume percentage(V10 Gy-V40 Gy)that normal brain tissue received 10 Gy-40 Gy also were observed and compared.Results:Compared with Planuniformity of IMRT,the Dmean of GTV of Planuniform increased-dose and Planincremental of IMRT increased by 10.13%and 17.9%,with statistically significant differences(t=13.680,12.771,P＜0.05).D50%increased by 8.9%and 10.8%,with statistically significant differences(t=15.190,9.929,P＜0.05).D2%increased by 15.2%and 46.4%,with statistically significant differences(t=52.320,8.746,P＜0.05).There were no statistically significant differences in normal brain tissue V10 Gy-V40 Gy among Planuniformity,Planuniform increased-dose and Planincremental of IMRT(P＞0.05).Compared with Planuniformity of VMAT,the Dmean of GTV of Planuniform increased-dose and Planincremental of VMAT increased by 10.53%and 21.23%,with statistically significant differences(t=18.641,15.461,P＜0.05),and D50%increased by 9.1%and 13.4%,with statistically significant differences(t=11.382,10.952,P＜0.05),and D2%increased by 16.4%and 48.8%,with statistically significant differences(t=56.471,8.685,P＜0.05),respectively.There were no statistically significant differences in normal brain tissue V10 Gy-V40 Gy among Planuniformity,Planuniform increased-dose and Planincremental of VMAT(P＞0.05).The normal brain tissue V20 Gy,V30 Gy and V40 Gy of Planuniform increased-dose and Planincremental of IMRT were respectively less than those of VMAT,and the differences of them between IMRT and VMAT were significant(tPlan uniform increased-dose=2.112,2.215,2.444,tPlan incremental=2.323,2.939,3.145,P＜0.05).There were no statistically significant difference in D2%,Dmean,and D50%between IMRT and VMAT(P＞0.05).Conclusion:On the premise of ensuring the safety of normal brain tissue at the edge of the target area,the synchronously increasing of the central dose of the target area will not significantly increase the dose for normal brain tissue.Both IMRT and VMAT can meet the requirements of increment in the inside of the target area,and VMAT has slightly better increment and higher efficiency within target area.The incremental of VMAT target area is slightly better,which also has better efficiency,while the enhancement effect of the dose of target area of Planincremental is better than that of the Planuniform increased-dose.The Plan incremental of VMAT is more suitable for HSRT treatment for BMs.