Objective To explore a standardized approach for indicating dosages in Chinese patent medicines(CPM)instructions.Methods A review of 1 378 classic formulas of traditional Chinese medicine in the 2020 edition of the Pharmaco-poeia of the People's Republic of China(Chinese Pharmacopoeia)was conducted,focusing on three aspects:overdose,different dosage forms,and inclusion of toxic herbs.Approximately 50 representative formulas were selected,and their daily dosage of herbal pieces and the corresponding dosage of individual herbs were calculated.These results were then compared and analyzed against the dosages stipulated in the Chinese Pharmacopoeia.Results ①Overdose:Among the 1 378 reviewed formulas,146(10.60％)were found to involve overdose.Specifically,eight formulas were identified as having an overdose of approximately 3 to 5 times the recommended dose,including Duanxueliu tablets and Zhixue Fumai mixture.②Dosage variations across dosage forms:Based on the conversion of the Ming-Qing dynasty dosage of one liang(Chinese traditional weight unit)to 36 g,the estimated dai-ly dose per component of crude herbs in Yinqiao powder was close to the upper limit specified in the Chinese Pharmacopoeia.The total daily dose of crude herbs(75.60 g)was approximately five times that of the Yinqiao powder(15 g)listed in the Chinese Pharmacopoeia and three times that of the Japanese Kampo Yinqiao powder(23.868 g).Significant differences in dosage were ob-served among Yinqiao Jiedu preparations of different dosage forms.In terms of total daily dose of crude herbs,granules(52.50 g)had the highest dose,approximately 3-5 times that of tablets/capsules(11.20 g),soft capsules(13.44 g),and powders(15.00 g),but still 30％ lower than the original prescription from ancient texts(75.6 g).Comparing the daily dose per compo-nent,granules fell within the Chinese Pharmacopoeia dose range,while tablets,capsules,soft capsules,and powders were all below the Chinese Pharmacopoeia dose.③Inclusion of toxic herbs:Among the 20 oral formulations containing Strychnos nux-vomica(Maqianzi)listed in the Chinese Pharmacopoeia,seven species exceeded the pharmacopoeial dose of 0.3-0.6 g per day.Notably,Shenjin Huoluo pill and Shufeng Dingtong pill exceeded the Chinese Pharmacopoeia dose by 1.5-2 times.The daily dose of strych-nosine in Shenjin Huoluo pill was 21.3 mg,exceeding the toxic dose range of 5-10 mg.Conclusions The dosage indications in CPM instructions must adhere to a unified format,clearly stating the daily dose per component of crude herbs,the total daily dose of crude herbs,and the total number of administrations under the"Dosage and Administration"section.For instance,"Yinqiao Jiedu granules,with a total daily dose equivalent to 60 g of crude herbs taken in four doses(including:Flos Lonicerae 10.71 g,Fructus Forsythiae 10.71 g,Radix Platycodi 6.43 g,Herba Menthae Haplocalycis 6.43 g,Fructus Arctii 6.43 g,Semen Sojae Preparatum 5.36 g,Radix Glycyrrhizae 5.36 g,Herba Lophatheri 4.29 g,Herba Schizonepetae 4.29 g)."For CPM containing tox-ic herbs,dosage indications are even more crucial,such as specifying the daily dose of Maqianzi powder/processed Maqianzi and strychnosine in formulations containing this herb.

Objective:To analyze the influencing factors of severe dengue fever patients in the early stage, construct a early prediction model for severe dengue fever, and evaluate it.Methods:A retrospective analysis was conducted to collect early clinical data of dengue fever patients admitted to the People's Hospital of Mengla County and the Third People's Hospital of Kunming in Yunnan Province from July to December 2023. The multifactor logistic regression was used to analyze the factors affecting the severe dengue fever patients, and Nomogram prediction model was used for visualization. Receiver operating characteristic (ROC) curve and calibration curve analysis were used to evaluate the model.Results:A total of 534 dengue fever patients were included, including 291 males and 243 females, aged (39.95 ± 15.69) years. Among them, there were 59 cases (11.05%) of severe dengue fever. The results of multifactor logistic regression analysis showed that age ( OR = 1.05, 95% CI: 1.02 - 1.08, P < 0.001), cardiovascular disease ( OR = 5.28, 95% CI: 2.08 - 13.40, P < 0.001), serous effusion ( OR = 4.34, 95% CI: 1.63 - 11.57, P = 0.003), aspartate aminotransferase ( OR = 1.03, 95% CI: 1.02 - 1.04, P < 0.001), lactate dehydrogenase ( OR = 1.00, 95% CI: 1.00 - 1.01, P = 0.001), and fibrinogen ( OR = 0.46, 95% CI: 0.28 - 0.76, P = 0.003) were independent influencing factors in the early stage of severe dengue fever. The area under the ROC curve of the Nomogram prediction model constructed from the above six variables was 0.96 (0.93 - 0.98). The calibration curve analysis results showed that the mean absolute error between the predicted values of the Nomogram prediction model and the actual observed values was 0.014. Conclusions:Age, cardiovascular disease, serous effusion, aspartate aminotransferase, lactate dehydrogenase, and fibrinogen are independent influencing factors in the early stage of severe dengue fever. The Nomogram prediction model established based on these variables has good predictive ability for severe dengue fever.

Objective To explore a standardized approach for indicating dosages in Chinese patent medicines(CPM)instructions.Methods A review of 1 378 classic formulas of traditional Chinese medicine in the 2020 edition of the Pharmaco-poeia of the People's Republic of China(Chinese Pharmacopoeia)was conducted,focusing on three aspects:overdose,different dosage forms,and inclusion of toxic herbs.Approximately 50 representative formulas were selected,and their daily dosage of herbal pieces and the corresponding dosage of individual herbs were calculated.These results were then compared and analyzed against the dosages stipulated in the Chinese Pharmacopoeia.Results ①Overdose:Among the 1 378 reviewed formulas,146(10.60％)were found to involve overdose.Specifically,eight formulas were identified as having an overdose of approximately 3 to 5 times the recommended dose,including Duanxueliu tablets and Zhixue Fumai mixture.②Dosage variations across dosage forms:Based on the conversion of the Ming-Qing dynasty dosage of one liang(Chinese traditional weight unit)to 36 g,the estimated dai-ly dose per component of crude herbs in Yinqiao powder was close to the upper limit specified in the Chinese Pharmacopoeia.The total daily dose of crude herbs(75.60 g)was approximately five times that of the Yinqiao powder(15 g)listed in the Chinese Pharmacopoeia and three times that of the Japanese Kampo Yinqiao powder(23.868 g).Significant differences in dosage were ob-served among Yinqiao Jiedu preparations of different dosage forms.In terms of total daily dose of crude herbs,granules(52.50 g)had the highest dose,approximately 3-5 times that of tablets/capsules(11.20 g),soft capsules(13.44 g),and powders(15.00 g),but still 30％ lower than the original prescription from ancient texts(75.6 g).Comparing the daily dose per compo-nent,granules fell within the Chinese Pharmacopoeia dose range,while tablets,capsules,soft capsules,and powders were all below the Chinese Pharmacopoeia dose.③Inclusion of toxic herbs:Among the 20 oral formulations containing Strychnos nux-vomica(Maqianzi)listed in the Chinese Pharmacopoeia,seven species exceeded the pharmacopoeial dose of 0.3-0.6 g per day.Notably,Shenjin Huoluo pill and Shufeng Dingtong pill exceeded the Chinese Pharmacopoeia dose by 1.5-2 times.The daily dose of strych-nosine in Shenjin Huoluo pill was 21.3 mg,exceeding the toxic dose range of 5-10 mg.Conclusions The dosage indications in CPM instructions must adhere to a unified format,clearly stating the daily dose per component of crude herbs,the total daily dose of crude herbs,and the total number of administrations under the"Dosage and Administration"section.For instance,"Yinqiao Jiedu granules,with a total daily dose equivalent to 60 g of crude herbs taken in four doses(including:Flos Lonicerae 10.71 g,Fructus Forsythiae 10.71 g,Radix Platycodi 6.43 g,Herba Menthae Haplocalycis 6.43 g,Fructus Arctii 6.43 g,Semen Sojae Preparatum 5.36 g,Radix Glycyrrhizae 5.36 g,Herba Lophatheri 4.29 g,Herba Schizonepetae 4.29 g)."For CPM containing tox-ic herbs,dosage indications are even more crucial,such as specifying the daily dose of Maqianzi powder/processed Maqianzi and strychnosine in formulations containing this herb.

Objective:To analyze the influencing factors of severe dengue fever patients in the early stage, construct a early prediction model for severe dengue fever, and evaluate it.Methods:A retrospective analysis was conducted to collect early clinical data of dengue fever patients admitted to the People's Hospital of Mengla County and the Third People's Hospital of Kunming in Yunnan Province from July to December 2023. The multifactor logistic regression was used to analyze the factors affecting the severe dengue fever patients, and Nomogram prediction model was used for visualization. Receiver operating characteristic (ROC) curve and calibration curve analysis were used to evaluate the model.Results:A total of 534 dengue fever patients were included, including 291 males and 243 females, aged (39.95 ± 15.69) years. Among them, there were 59 cases (11.05%) of severe dengue fever. The results of multifactor logistic regression analysis showed that age ( OR = 1.05, 95% CI: 1.02 - 1.08, P < 0.001), cardiovascular disease ( OR = 5.28, 95% CI: 2.08 - 13.40, P < 0.001), serous effusion ( OR = 4.34, 95% CI: 1.63 - 11.57, P = 0.003), aspartate aminotransferase ( OR = 1.03, 95% CI: 1.02 - 1.04, P < 0.001), lactate dehydrogenase ( OR = 1.00, 95% CI: 1.00 - 1.01, P = 0.001), and fibrinogen ( OR = 0.46, 95% CI: 0.28 - 0.76, P = 0.003) were independent influencing factors in the early stage of severe dengue fever. The area under the ROC curve of the Nomogram prediction model constructed from the above six variables was 0.96 (0.93 - 0.98). The calibration curve analysis results showed that the mean absolute error between the predicted values of the Nomogram prediction model and the actual observed values was 0.014. Conclusions:Age, cardiovascular disease, serous effusion, aspartate aminotransferase, lactate dehydrogenase, and fibrinogen are independent influencing factors in the early stage of severe dengue fever. The Nomogram prediction model established based on these variables has good predictive ability for severe dengue fever.

Objective To investigate the mechanism of Guizhi Tongluo Tablets(GZTLP)on improving atherosclerosis in APOE knockout mice by regulating neutrophil extracellular trapping nets(NETs).Methods After modeling,24 APOE knockout mice aged 8 weeks were randomly divided into 4 groups:GZTLP high-dose group,low-dose group,model control group and normal control group,with 6 mice in each group.GZTLP was given 1.87 mg·g-1 and 0.47 mg·g-1 intragastric administration in high-dose group and low-dose group,respectively.The normal control group and model control group were given 0.9%sodium chloride solution intragastric administration for 6 weeks,and the lipid plaque deposition in aorta was observed by gross oil red O staining.Lipid deposition in aortic root was observed by oil red O staining.The pathological changes of lipid plaques in aortic root were observed by HE staining.The levels of interleukin-1β(IL-1β)and tumor necrosis factor α(TNF-α)in peripheral blood of mice were detected by enzyme-linked immunosorbent assay(ELISA).The expression of lymphocyte antigen 6G(Ly6G),myeloperoxidase(MPO)and citrulinated histone(Cit-H3)in plaques of the aortic arch and the colocalization of Ly6G,MPO and Cit-H3 were detected by immunofluorescence assay.Results Compared with the normal control group,the aorta of mice in the model control group showed serious lipid plaque deposition,morphological damage,and a large number of inflammatory cells infiltration,the contents of serum inflammatory factors IL-1β and TNF-α were increased,and the protein expressions of Ly6G,Cit-H3 and MPO were significantly increased.Compared with model control group,GZTLP group reduced the amount of lipid plaque deposition in aorta,the arrangement of aortic cells was more regular,the inflammatory cell infiltration was improved,and the contents of serum inflammatory factors IL-1β and TNF-α were significantly decreased(P<0.05).The colocalization and the protein expression of Ly6G,MPO and Cit-H3 were significantly decreased in aortic tissues(P<0.01).Conclusions GZTLP can improve atherosclerosis,and its mechanism may be related to the inhibition of neutrophil extracellular trapping nets.

Objective To explore the mechanism of Nuanxinkang Powder(aka.NXK,composed of Ginseng Radix et Rhizoma Rubra and Ilex Pubescens Radix)on improving ventricular remodeling in post-infarction mice based on the"metabolic-inflammatory"network regulating macrophage polarization.Methods ①Thirty C57BL/6J male mice were randomly divided into three groups:sham-operation group,model group,and NXK group(1.65 g·kg-1),with 10 mice in each group;the mouse model of myocardial infarction was replicated using left anterior descending coronary artery ligation;and the drug was administered by gavage once a day for 4 consecutive weeks.Masson staining was used to detect collagen deposition in myocardial tissue;ultrasound was used to detect cardiac function in mice:left ventricular ejection fraction(LVEF),left ventricular anterior wall thickness at end-systole(LVAWS)and left ventricular anterior wall thickness at end-diastole(LVAWD);flow cytometry was used to detect distribution of cardiac macrophages in mice;qPCR was used to detect mRNA expressions of lactate dehydrogenase A(LDHA),carnitine palmitoyltransferase 1(CPT-1),glucose transport protein 4(GLUT4),isocitrate dehydrogenase(IDH),and succinate dehydrogenase(SDHa)in heart tissue.②NXK was given 1.15 g·kg-1 NXK suspension to rats by gavage twice a day for 5 consecutive days to prepare NXK-containing serum.Lipopolysaccharide(LPS)-induced RAW 264.7 cells were used to construct a pro-inflammatory macrophage model.The cells were grouped into the following groups:blank serum control group(medium containing 5%blank serum+5%fetal bovine serum),NXK drug-containing serum group(medium containing 5%NXK drug-containing serum+5%fetal bovine serum),lipopolysaccharide group(medium containing 5%blank serum+5%fetal bovine serum+200 μg·mL-1 lipopolysaccharide),NXK drug-containing serum+ lipopolysaccharide group(medium containing 5%NXK drug-containing serum+5%fetal bovine serum+200 μ g·mL-1 lipopolysaccharide),all the groups were intervened for 16 hours.Glycolysis stress test was used to detect the level of glycolysis in RAW 264.7 cells;qPCR was used to detect the mRNA expression of mitochondrial pyruvate carrier(MPC1)in RAW 264.7 cells;and MitoSox Red fluorescent staining was used to detect the level of oxidative stress damage in mitochondria of RAW 264.7 cells.Results ①Compared with the sham-operation group,the blue-stained area of cardiac collagen fibres in mice of the model group was significantly increased,accompanied by thinning of the ventricular wall and enlargement of the left ventricular cavity;cardiac function indexes,such as LVEF,LVAWS,LVAWD,etc.,were all significantly reduced(P＜0.01,P＜0.001);the mRNA expressions of LDHA and CPT-1 were significantly up-regulated in the cardiac tissues of mice(P＜0.05),and the mRNA expressions of GLUT4,IDH and SDHa were significantly down-regulated(P＜0.05,P＜0.01),and CD86 staining positive cell was significantly increased(P＜0.001).Compared with the model group,mice in the NXK group showed a significant decrease in cardiac collagen fiber deposition and an increase in the thickness of the ventricular wall;cardiac function indexes such as LVEF,LVAWS and LVAWD were significantly increased(P＜0.05,P＜0.01,P＜0.001);and the mRNA expressions of LDHA and CPT-1 in the cardiac tissues of the mice were significantly down-regulated(P＜0.01,P＜0.001),mRNA expressions of GLUT4,SDHa and IDH were significantly up-regulated(P＜0.01),and the number of CD86 positive cells was significantly reduced(P＜0.001).②Compared with the blank serum control group,the cytosolic glycolysis level and ROS level of macrophages in the NXK serum-containing group did not change significantly(P＞0.05),whereas the glycolysis level and ROS level of macrophages in the lipopolysaccharide group were significantly increased(P＜0.01),and the mRNA expression of MPC1 was significantly down-regulated(P＜0.001).Compared with the lipopolysaccharide group,the macrophage glycolysis level and ROS level were significantly reduced in the NXK serum-containing + lipopolysaccharide group(P＜0.05,P＜0.01),and mRNA expression of MPC1 was significantly up-regulated(P＜0.001).Conclusion NXK can reduce myocardial fibrosis and ventricular remodeling after myocardial infarction and improve cardiac function in mice,and its mechanism may be related to the down-regulation of mRNA expression of LDHA in cardiac tissues,the up-regulation of mRNA expression of GLUT4,the improvement of cardiac glucose uptake after myocardial infarction,the inhibition of pro-inflammatory macrophage glycolysis,the increase in the expressions of SDHa and IDH to alleviate the accumulation of succinate and citrate,and the reduction of reactive oxygen species(ROS)generation,thereby reducing pro-inflammatory macrophage hyperpolarisation.

The solute carrier family 12(SLC12)of cation-chloride cotransporters(CCCs)comprises potassium chlo-ride cotransporters(KCCs,e.g.KCC1,KCC2,KCC3,and KCC4)-mediated Cl-extrusion,and sodium po-tassium chloride cotransporters(N[K]CCs,NKCC1,NKCC2,and NCC)-mediated Cl-loading.The CCCs play vital roles in cell volume regulation and ion homeostasis.Gain-of-function or loss-of-function of these ion transporters can cause diseases in many tissues.In recent years,there have been considerable ad-vances in our understanding of CCCs'control mechanisms in cell volume regulations,with many tech-niques developed in studying the functions and activities of CCCs.Classic approaches to directly measure CCC activity involve assays that measure the transport of potassium substitutes through the CCCs.These techniques include the ammonium pulse technique,radioactive or nonradioactive rubidium ion uptake-assay,and thallium ion-uptake assay.CCCs'activity can also be indirectly observed by measuring y-aminobutyric acid(GABA)activity with patch-clamp electrophysiology and intracellular chloride con-centration with sensitive microelectrodes,radiotracer 36Cl-,and fluorescent dyes.Other techniques include directly looking at kinase regulatory sites phosphorylation,flame photometry,22Na+uptake assay,structural biology,molecular modeling,and high-throughput drug screening.This review sum-marizes the role of CCCs in genetic disorders and cell volume regulation,current methods applied in studying CCCs biology,and compounds developed that directly or indirectly target the CCCs for disease treatments.

Objective To compare the clinical application value of the real-time fluorescence PCR method and flow cytometry method for detecting human leukocyte antigen B27(HLA-B27).Methods Blood HLA-B27 level in 225 patients with suspected ankylosing spondylitis was detected by using real-time fluorescence PCR method and flow cytometry method.The detection results were compared and analyzed between the two methods.Results The results of 95.1% sample were identical detected by two methods without statistically significant difference(P>0.05).Taken the results of flow cytometry as reference, the sensitivity of real-time fluorescence PCR for detecting HLA-B27 was 94%, the specificity was 96%.Gene sequencing was performed if results of a sample detected by two methods were different, which was identical with the result detected by real-time fluorescence PCR.Conclusion Both methods for detecting HLA-B27 all have high sensitivity and specificity.Real-time fluorescence PCR method is more superior to the flow cytometry method in the results accuracy.

Background Our previous studies found that mesenchymal stem cells (MSCs) can ameliorate experimental autoimmune uveitis (EAU) and reduce tissue impairment.Its mechanism is still pending.Objective This study was performed to investigate the effects of MSCs on T cell subsets and antigen presenting cells (APCs) in EAU rats.Methods MSCs were isolated from bone marrow of six male Wistar rats and cultured by plastic adherence method.Twelve female Lewis rats were assigned randomly into MSCs group and PBS group.EAU rat model was induced by immunization with 200 μl emulsion containing 30 μg interphotoreceptor retinoid-binding protein (IRBP) 1177-1191 polypeptide fragment R16 and complete Freund adjuvant (CFA).The eye manifestations of the rats were observed and scored under the slit lamp microscope after modeling.The R16-immunized rats were treated intravenously with 5×106/ml MSCs for 3 consecutive days from day 9 to 11 after modeling in the MSCs group,and the equivalent volume of PBS was used with the same way in the PBS group.Fifteen days after modeling,the spleens and draining lymph nodes were collected to evaluate the proportion of interferon-γ (IFN-γ) positive CD4+ T cells,interleukin-17 (IL-17)positive CD4+ T cells and forkhead helix transcription factor p3 (Foxp3) positive CD4+ T cells by flow cytometry.The T cells and APCs from the different groups were cocultured and divided into PBS cocultured group,MSCs cocultured group, PBS-MSCs cross-cultured group and MSCs-PBS cross-cultured group under the stimulation of R16 at the concentration of 0.3,1.0 or 10.0 μg/ml, and the proliferation indexes of the T cells in different groups were assayed by 5-bromodeoxyuridine (BrdU) Elisa kit.The use of experimental animals complied with the regulations on the management of experimental animals promulgated by the national science and technology commission.Results The ocular surface inflammatory scores of 11,12,13 and 14 days after modeling in the MSCs group were significantly lower than that in the PBS group (t=3.825,5.100,4.250,3.400, all at P＜0.05).Compared with the PBS group, the proportions of IFN-γ positive CD4+ T cells in spleen and draining lymph notes were considerably decreased in the MSCs group (t =5.651,4.376, both at P＜0.05) , so were the IL-17+ CD4+ T cells (t =3.300,4.925, both at P＜0.05).However,the proportions of Foxp3 + CD4+ T cells in spleen and draining lymph notes were statistically raised in the MSCs group compared with the PBS group (t =-5.172,-2.825,both at P＜0.05).The proliferation index of T cells increased with the rise of R16 dose in the PBS cocultured group, and the proliferation indexes were all declined in the MSCs cocultured group compared with the PBS cocultured group under the stimulation of 0.3,1.0 and 10.0 μg/ml of R16 (P =0.027,0.000,0.000).In addition, significant reduces of proliferation indexes of T cells were seen in the PBS-MSCs cross-cultured group and MSCs-PBS cross-cultured group in comparison with the PBS cocultured group when stimulated by 1.0 μg/ml and 10.0 μg/ml R16 (1.0 μg/ml R16 : P =0.001,0.000;10.0 μg/ml R16:P=0.000,0.000).Conclusions MSCs can ameliorate EAU by inhibiting the functions of antigen-specific T cells and APCs and up-regulating T regulatory cells in EAU rats.