OBJECTIVE: Therapeutic angiogenesis has become a promising approach for treating ischemic heart disease (IHD). The present study aims to investigate the effects of Qishen Granule (QSG) on angiogenesis in myocardial ischemia (MI) and the potential mechanism. METHODS: In vivo study was conducted on rat model of myocardial infarction. QSG was performed daily at a dose of 2.352 g/kg for four weeks. Cardiac function was assessed by echocardiogram and pro-angiogenic effects were evaluated by Laser Doppler and CD31 expression. Oxygen-glucose deprivation (OGD) was applied in cultured human umbilical vein endothelial cells (HUVECs). Cell viability, wound healing and tube formation assay were used to test functions of HUVECs. ELISA and Western blots were used to assess protein expressions of bone morphogenetic protein 2-delta-like 4-notch homolog 1 (BMP2-Dll4-Notch1) signaling pathway. RESULTS: The results showed that QSG improved heart function, cardiac blood flow and microvessel density in myocardial ischemic rats. In vitro, QSG protected HUVECs by promoting the cell viability and tube formation. QSG upregulated bone morphogenetic protein-2 (BMP2) and downregulated delta-like 4 (Dll4) and notch homolog 1 (Notch1) expressions both in rats and HUVECs. CONCLUSION QSG protected against MI by promoting angiogenesis through BMP2-Dll4-Notch1 pathway. BMP2 might be a promising therapeutic target for IHD.

In recent years,chemical imaging technology has found increasingly broad applications in pharmaceuti-cal analysis,encompassing component distribution analysis,assessment of mixing uniformity,content uniformity e-valuation,polymorphism identification,and studies of drug delivery mechanisms.Chemical imaging has been pro-gressively recognized as an efficient tool for real-time quality control and online analysis.This article provides an o-verview of the chemical imaging guidelines in the Chinese,United States,European pharmacopoeias.The discus-sion aims to promote the application of multi-information fusion technologies in pharmaceutical analysis,enhance regulatory efficiency,and further advance the high-quality development of the pharmaceutical industry in China.

Objective To investigate the protective effect of hederagenin(HDG)on cisplatin(Cis)-induced acute kidney injury(AKI)in mice and its potential mechanism.Methods 24 male C57BL/6J mice were randomly divided into a control group,AKI model group,HDG low-dose group,and HDG high-dose group,with six mice in each group.AKI model was established by intraperitoneal injection of 20 mg/kg cisplatin(Cis).The HDG low-dose and HDG high-dose groups were given 20,40 mg/kg HDG by intragastric administration,respectively,and samples were collected 3 days later.The kidneys of the mice were collected for hematoxylin-eosin(HE)and periodic-acid-schiff(PAS)staining to evaluate the kidney pathology,and serum was collected to detect changes in serum creatinine(Scr)and blood urea nitrogen(BUN).The expression of p-P65,P65,IL-6,TNF-α,IL-1β,and other inflammatory-related proteins was detected by Western Blot.A TCMK1(renal tubular epithelial cell)inflammatory cell model was established by Cis(200 ng/mL)stimulation in vitro.Blank group,Cis model group,HDG low-dose group,HDG high-dose group,Axin2 overexpression group,HDG+Axin2 overexpression group were set up.In the Axin2-overexpression group,the expression of p-P65,P65,IL-6,TNF-α,IL-1β,Axin2,and AREG was detected among total cell proteins.Results Compared with the control group,AKI model mice exhibited significantly elevated serum creatinine and blood urea nitrogen levels(P<0.05),accompanied by pathological alterations including vacuolar degeneration of renal tubules,inflammatory cell infiltration,and glycogen deposition,and the expression of inflammation-related proteins(p-P65,TNF-α,IL-6,IL-1β)and Axin2 was markedly upregulated in AKI mice(P<0.05).HDG treatment induced a dose-dependent reduction in serum creatinine and blood urea nitrogen levels(high-dose>low-dose,P<0.05),alleviated renal histopathological damage,and concurrently suppressed the expression of these inflammatory mediators and Axin2(P<0.05).HDG was confirmed that dose-dependently inhibited Cis-induced upregulation of Axin2,and inflammatory cytokines in vitro experiments.Transcriptome sequencing revealed that Axin2 overexpression significantly increased amphiregulin(AREG)expression(P<0.05).Mechanistically,HDG reduced p-P65 phosphorylation by suppressing the Axin2/AREG axis(P<0.05),while Axin2 overexpression abolished the protective effects of HDG against Cis-induced renal tubular cell injury.Conclusions HDG protects against renal injury in AKI mice by reducing inflammation through the inhibition of Axin2/AREG axis activation.

AIM:To explore the potential of Huangqi sanqi mixture(AP)in improving renal fi-brosis by performing transcriptome sequencing of the kidneys of the unilateral ureteral ligation mouse group and the Huangqi sanqi mixture inter-vention group,and using bioinformatics to verify the signitficantly different lncRNAs mechanism.METHODS:Twenty-four C57 mice were divided in-to sham operation group,renal fibrosis group,Huangqi sanqi mixture intervention group(3.944 g/kg)and irbesartan positive control intervention group,with 6 mice in each group.A mouse model of renal fibrosis was established by unilateral ure-teral ligation(UUO).The animals were given intra-gastric administration after operation,and the ani-mals were sacrificed and the specimens were col-lected after seven consecutive days of administra-tion.The changes of Huangqi sanqi mixture on re-nal fibrosis pathological damage were analyzed by HE and Masson staining,and the protein levels of Fn and α-SMA in renal tissue of each group were detected by Western blot and immunohistochemis-try to evaluate the alleviating effect of Huangqi san-qi mixture on renal fibrosis.Subsequently,lncRNA expression information was obtained by transcrip-tome sequencing,and Quantitative Real-time PCR(qPCR)was performed after data quality,GO en-richment and differential lncRNA were analyzed.According to the differential lncRNA and target analysis results obtained by sequencing,lncRNA Gm51500/Adam12 was overexpressed in vitro,and its mechanism in the protection of renal fibrosis by Huangqi sanqi mixture was studied by immunohis-tochemistry,immunofluorescence staining and qP-CR verification.RESULTS:Compared with the mod-el group,the renal fibrosis of the mice in the Huangqi sanqi mixture intervention group was sig-nificantly reduced,and the protein levels of α-SMA and Fn and the expression of lncRNA in the renal tis-sue were significantly down-regulated(P＜0.000 1).Three lncRNAs were screened and verified to in-crease in the model group and significantly de-crease after AP intervention,namely lncRNA Gm29994,Gm51500 and Gm35391.Target analysis showed that lncRNA Gm51500 had the most signifi-cant relationship with Adam12.The results of ani-mal experiments showed that Adam12 was highly expressed in the kidney of UUO mice and was sig-nificantly inhibited after AP intervention.Subse-quent cell experiments confirmed that overexpres-sion of lncRNA Gm51500 could up-regulate TGF-β-induced renal tubular cell fibrosis and Adam12 ex-pression.Cell recovery experiments confirmed that Adam12 overexpression reversed the inhibitory ef-fect of AP on renal tubular cell injury and fibrosis.CONCLUSION:Huangqi sanqi mixture can improve renal fibrosis.Based on transcriptomic sequencing,lncRNA Gm51500/Adam12 axis may be a potential target for Huangqi sanqi mixture to improve renal fibrosis.

Objective To investigate the protective effect of hederagenin(HDG)on cisplatin(Cis)-induced acute kidney injury(AKI)in mice and its potential mechanism.Methods 24 male C57BL/6J mice were randomly divided into a control group,AKI model group,HDG low-dose group,and HDG high-dose group,with six mice in each group.AKI model was established by intraperitoneal injection of 20 mg/kg cisplatin(Cis).The HDG low-dose and HDG high-dose groups were given 20,40 mg/kg HDG by intragastric administration,respectively,and samples were collected 3 days later.The kidneys of the mice were collected for hematoxylin-eosin(HE)and periodic-acid-schiff(PAS)staining to evaluate the kidney pathology,and serum was collected to detect changes in serum creatinine(Scr)and blood urea nitrogen(BUN).The expression of p-P65,P65,IL-6,TNF-α,IL-1β,and other inflammatory-related proteins was detected by Western Blot.A TCMK1(renal tubular epithelial cell)inflammatory cell model was established by Cis(200 ng/mL)stimulation in vitro.Blank group,Cis model group,HDG low-dose group,HDG high-dose group,Axin2 overexpression group,HDG+Axin2 overexpression group were set up.In the Axin2-overexpression group,the expression of p-P65,P65,IL-6,TNF-α,IL-1β,Axin2,and AREG was detected among total cell proteins.Results Compared with the control group,AKI model mice exhibited significantly elevated serum creatinine and blood urea nitrogen levels(P<0.05),accompanied by pathological alterations including vacuolar degeneration of renal tubules,inflammatory cell infiltration,and glycogen deposition,and the expression of inflammation-related proteins(p-P65,TNF-α,IL-6,IL-1β)and Axin2 was markedly upregulated in AKI mice(P<0.05).HDG treatment induced a dose-dependent reduction in serum creatinine and blood urea nitrogen levels(high-dose>low-dose,P<0.05),alleviated renal histopathological damage,and concurrently suppressed the expression of these inflammatory mediators and Axin2(P<0.05).HDG was confirmed that dose-dependently inhibited Cis-induced upregulation of Axin2,and inflammatory cytokines in vitro experiments.Transcriptome sequencing revealed that Axin2 overexpression significantly increased amphiregulin(AREG)expression(P<0.05).Mechanistically,HDG reduced p-P65 phosphorylation by suppressing the Axin2/AREG axis(P<0.05),while Axin2 overexpression abolished the protective effects of HDG against Cis-induced renal tubular cell injury.Conclusions HDG protects against renal injury in AKI mice by reducing inflammation through the inhibition of Axin2/AREG axis activation.

In recent years,chemical imaging technology has found increasingly broad applications in pharmaceuti-cal analysis,encompassing component distribution analysis,assessment of mixing uniformity,content uniformity e-valuation,polymorphism identification,and studies of drug delivery mechanisms.Chemical imaging has been pro-gressively recognized as an efficient tool for real-time quality control and online analysis.This article provides an o-verview of the chemical imaging guidelines in the Chinese,United States,European pharmacopoeias.The discus-sion aims to promote the application of multi-information fusion technologies in pharmaceutical analysis,enhance regulatory efficiency,and further advance the high-quality development of the pharmaceutical industry in China.

AIM:To explore the potential of Huangqi sanqi mixture(AP)in improving renal fi-brosis by performing transcriptome sequencing of the kidneys of the unilateral ureteral ligation mouse group and the Huangqi sanqi mixture inter-vention group,and using bioinformatics to verify the signitficantly different lncRNAs mechanism.METHODS:Twenty-four C57 mice were divided in-to sham operation group,renal fibrosis group,Huangqi sanqi mixture intervention group(3.944 g/kg)and irbesartan positive control intervention group,with 6 mice in each group.A mouse model of renal fibrosis was established by unilateral ure-teral ligation(UUO).The animals were given intra-gastric administration after operation,and the ani-mals were sacrificed and the specimens were col-lected after seven consecutive days of administra-tion.The changes of Huangqi sanqi mixture on re-nal fibrosis pathological damage were analyzed by HE and Masson staining,and the protein levels of Fn and α-SMA in renal tissue of each group were detected by Western blot and immunohistochemis-try to evaluate the alleviating effect of Huangqi san-qi mixture on renal fibrosis.Subsequently,lncRNA expression information was obtained by transcrip-tome sequencing,and Quantitative Real-time PCR(qPCR)was performed after data quality,GO en-richment and differential lncRNA were analyzed.According to the differential lncRNA and target analysis results obtained by sequencing,lncRNA Gm51500/Adam12 was overexpressed in vitro,and its mechanism in the protection of renal fibrosis by Huangqi sanqi mixture was studied by immunohis-tochemistry,immunofluorescence staining and qP-CR verification.RESULTS:Compared with the mod-el group,the renal fibrosis of the mice in the Huangqi sanqi mixture intervention group was sig-nificantly reduced,and the protein levels of α-SMA and Fn and the expression of lncRNA in the renal tis-sue were significantly down-regulated(P＜0.000 1).Three lncRNAs were screened and verified to in-crease in the model group and significantly de-crease after AP intervention,namely lncRNA Gm29994,Gm51500 and Gm35391.Target analysis showed that lncRNA Gm51500 had the most signifi-cant relationship with Adam12.The results of ani-mal experiments showed that Adam12 was highly expressed in the kidney of UUO mice and was sig-nificantly inhibited after AP intervention.Subse-quent cell experiments confirmed that overexpres-sion of lncRNA Gm51500 could up-regulate TGF-β-induced renal tubular cell fibrosis and Adam12 ex-pression.Cell recovery experiments confirmed that Adam12 overexpression reversed the inhibitory ef-fect of AP on renal tubular cell injury and fibrosis.CONCLUSION:Huangqi sanqi mixture can improve renal fibrosis.Based on transcriptomic sequencing,lncRNA Gm51500/Adam12 axis may be a potential target for Huangqi sanqi mixture to improve renal fibrosis.

Objective To establish the method for simultaneous determination of six index components in the water extract of Weile Prescription;To optimize the water extraction process.Methods UPLC-MS/MS was used with Waters CORTECS C18 column(2.1 mm×100 mm,1.6 μm)as the chromatographic conditions;the mobile phase was 0.1%formic acid water-acetonitrile with gradient elution;the flow rate was 0.25 mL/min;the column temperature was 40℃;the sample volume was 2 μL.Electrospray negative ion source,positive and negative ion switching multi-reaction monitoring(MRM)mode were detected.Taking the content of six index components(gallic acid,vitexin,paeoniflorin,naringin,hesperidin and glycyrrhizic acid)and extraction rate as evaluation indexes,the weight coefficient of each index was determined by G1-entropy weight method,and the optimum parameters of extraction process were determined by orthogonal experiment design with the amount of water,extraction time and extraction times as investigation factors.Results There was a good linear relationship of the six components in the water extract of Weile Prescription in the concentration range(r>0.999),and the average recovery rate was 96.83%-102.56%,RSD<4.0%.The best technological parameters were as follows:Chinese decoction pieces were soaked in 12 times of water for 2 h,and extracted twice,each time for 1.5 h.Conclusion The UPLC-MS/MS method established in the study for simultaneous determination of six components in Weile Prescription is rapid,simple and sensitive,and the optimized extraction process is stable and feasible,which provides experimental basis for the development and research of the preparation.

Objective To explore the application low of stimulation parameters of transcutaneous electrical acupoint stimulation(TEAS)and transcranial direct current stimulation(tDCS)for post-stroke movement disorders based on data mining.Methods The relevant clinical research literature was retrieved from CNKI,Wanfang Data,VIP,CBM,PubMed and Web of Science from January 2000 to May 2023.A database was set up after quality assessment.Frequency analysis,association rules and complex network analysis were used to explore the application law of core acupoints and electrical stimulation parameters.Results A total of 79 articles were included and 128 groups of data were contained.For TEAS,the core acupoints included Waiguan(TE5),Shousanli(LI10),Zusanli(ST36),Hegu(LI4),Neiguan(PC6),Yanglingquan(GB34),etc.,while the most commonly used acupoint combinations of upper limb and lower limb were Shousanli(LI10)-Waiguan(TE5)and Yanglingquan(GB34)-Zusanli(ST36).Among the electrical stimulation parameters of TEAS,the frequencies used vary widely,and 100 Hz was most commonly used,while 2 Hz TEAS was also mainly used for stimulating acupoints located on upper limbs in the treatment of flaccid paralysis.The application of other electrical stimulation parameters was relatively consistent.The bidirectional symmetrical square-wave with 200-250 μs pulse-width was used in majority of studies.The stimulus intensity was mostly determined by patient tolerance.For tDCS,stimulation electrodes were often positioned on the projection of the primary M1,and the safe stimulus intensity was mostly set as 1 to 2 mA.Conclusion In the treatment of post-stroke movement disorders,appropriate acupoints and electrical stimulation parameters of TEAS should be determined on the muscle strength and muscle tension of stroke patients at different stages after stroke,particularly the selection of electric stimulating frequency.

Bladder cancer remains the 10th most common cancer worldwide. In recent years, metformin has been found to have potential anti-bladder cancer activ-ity while high concentration of IC50 at millimolar level is needed, which could not be reached by regular oral administration route. Thus, higher efficient agent is urgently demanded for clinically treating bladder cancer. Here, by conjugating artesunate to metformin, a novel artesunate-metformin dimer triazine derivative AM2 was designed and synthesized. The inhibitory effect of AM2 on bladder cancer cell line T24 and the mechanism underlying was determined. Anti-tumor activity of AM2 was assessed by MTT, cloning formation and wound healing assays. Decreasing effect of AM2 on lipogenesis was determined by oil red O staining. The protein expressions of Clusterin, SREBP1 and FASN in T24 cells were evaluated by Western blotting. The results show that AM2 significantly inhibited cell proliferation and migration at micromolar level, much higher than parental metformin. AM2 reduced lipogenesis and down-regulated the expressions of Clusterin, SREBP1 and FASN. These results suggest that AM2 inhibits the growth of bladder cancer cells T24 by inhibiting cellular lipogenesis associated with the Clusterin/SREBP1/FASN signaling pathway.