Objective To study the transcriptional expression of Chlorophyti Laxi R.Br.using high-throughput sequencing technology.Methods Total RNA of Chlorophtum laxum R.Br.was extracted,followed by library construction,sequencing,and de novo assembly to obtain unigene sequences.These sequences were then annotated and compared to acquire genetic information of the Chlorophtum laxum R.Br.transcriptome.Results A total of 29 325 unigene were identified,with an average length of 1 026 bp and an N50 of 1 697 bp.Among them,20 338 unigene were functionally annotated in at least one database,with Chlorophtum laxum R.Br.showing the highest sequence similarity to Asparagus officinalis.In the Kyoto Encyclopedia of Genes and Genomes(KEGG)database,13 392 unigene were annotated,including genes involved in flavonoid biosynthesis and ubiquinone and other terpenoid-quinone biosynthesis pathways.Thirty-five upstream genes related to Chlorophtum laxum R.Br.saponin biosynthesis were identified.In the Gene Ontology(GO)database,16 728 unigene were annotated,covering cellular anatomical entities,binding,and biological processes.Using the Microsatellite Identification Tool(MISA),11 486 assembled unigene longer than 1 000 bp were analyzed for simple sequence repeats(SSRs),resulting in the identification of 5 178 SSR loci,for which primers were designed using Primer 3.0.By comparing Chlorophtum laxum R.Br.unigene with the transcription factor database,657 transcription factors,including bHLH,MYB,and WRKY,were identified.Conclusion The transcriptome data provide a foundation for studying the biosynthetic pathways and functional genes of medicinal active components in Chlorophtum laxum R.Br.,and also contribute to the conservation and development of its resources.

This study aims to reveal the effect and mechanism of Gentianella turkestanorum total extract(GTI) in ameliorating non-alcoholic steatohepatitis(NASH). UPLC-Q-TOF-MS was employed to identify the chemical components in GTI. SwissTarget-Prediction, GeneCards, OMIM, and TTD were utilized to screen the targets of GTI components and NASH. The common targets shared by GTI components and NASH were filtered through the STRING database and Cytoscape 3.9.0 to identify core targets, followed by GO and KEGG enrichment analysis. AutoDock was used for molecular docking of key components with core targets. A mouse model of NASH was established with a methionine-choline-deficient high-fat diet. A 4-week drug intervention was conducted, during which mouse weight was monitored, and the liver-to-brain ratio was measured at the end. Hematoxylin-eosin staining, Sirius red staining, and oil red O staining were employed to observe the pathological changes in the liver tissue. The levels of various biomarkers, including aspartate aminotransferase(AST), alanine aminotransferase(ALT), hydroxyproline(HYP), total cholesterol(TC), triglycerides(TG), low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione(GSH), in the serum and liver tissue were determined. RT-qPCR was conducted to measure the mRNA levels of interleukin 1β(IL-1β), interleukin 6(IL-6), tumor necrosis factor α(TNF-α), collagen type I α1 chain(COL1A1), and α-smooth muscle actin(α-SMA). Western blotting was conducted to determine the protein levels of IL-1β, IL-6, TNF-α, and potential drug targets identified through network pharmacology. UPLC-Q-TOF/MS identified 581 chemical components of GTI, and 534 targets of GTI and 1 157 targets of NASH were screened out. The topological analysis of the common targets shared by GTI and NASH identified core targets such as IL-1β, IL-6, protein kinase B(AKT), TNF, and peroxisome proliferator activated receptor gamma(PPARG). GO and KEGG analyses indicated that the ameliorating effect of GTI on NASH was related to inflammatory responses and the phosphoinositide 3-kinase(PI3K)/AKT pathway. The staining results demonstrated that GTI ameliorated hepatocyte vacuolation, swelling, ballooning, and lipid accumulation in NASH mice. Compared with the model group, high doses of GTI reduced the AST, ALT, HYP, TC, and TG levels(P<0.01) while increasing the HDL-C, SOD, and GSH levels(P<0.01). RT-qPCR results showed that GTI down-regulated the mRNA levels of IL-1β, IL-6, TNF-α, COL1A1, and α-SMA(P<0.01). Western blot results indicated that GTI down-regulated the protein levels of IL-1β, IL-6, TNF-α, phosphorylated PI3K(p-PI3K), phosphorylated AKT(p-AKT), phosphorylated inhibitor of nuclear factor kappa B alpha(p-IκBα), and nuclear factor kappa B(NF-κB)(P<0.01). In summary, GTI ameliorates inflammation, dyslipidemia, and oxidative stress associated with NASH by regulating the PI3K/AKT/NF-κB signaling pathway.
Animals ; Non-alcoholic Fatty Liver Disease/genetics* ; Mice ; Network Pharmacology ; Male ; Drugs, Chinese Herbal/administration & dosage* ; Chromatography, High Pressure Liquid ; Liver/metabolism* ; Mice, Inbred C57BL ; Humans ; Mass Spectrometry ; Tumor Necrosis Factor-alpha/metabolism* ; Disease Models, Animal ; Molecular Docking Simulation

OBJECTIVE To investigate the targeting effect of folic acid-modified crebanine nanoparticles （FA-Cre@PEG- PLGA NPs， hereinafter referred to as “NPs”） combined with ultrasound irradiation on M109 cells in vitro and in vivo after administration， and explore the anti-tumor mechanism. METHODS CCK-8 assay was used to detect the inhibitory effect of NPs combined with ultrasound irradiation on the proliferation of M109 cells， and the best ultrasound time was selected. Using human lung cancer A549 cells as a control， the targeting of NPs combined with ultrasound irradiation to M109 cells was evaluated by free folic acid blocking assay and cell uptake assay. The effects of NPs combined with ultrasound irradiation on the migration， invasion， apoptosis， cell cycle and reactive oxygen species （ROS） levels of M109 cells were detected by cell scratch test， Transwell chamber test and flow cytometry at 1 h after 958401536@qq.com administration； the changes of mitochondrial membrane potential （MMP） were observed by fluorescence inverted microscope. A mouse subcutaneous tumor model of M109 cells was constructed， and the in vivo tumor targeting of NPs combined with ultrasound irradiation was investigated by small animal in vivo imaging technology. RESULTS NPs combined with ultrasound irradiation could significantly inhibit the proliferation of M109 cells， and the optimal ultrasound time was 1 h after administration. The free folic acid could antagonize the inhibitory effect of NPs on the proliferation of M109 cells， and combined with ultrasound irradiation could partially reverse this antagonism. Compared with A549 cells， the uptake rate of NPs in M109 cells was significantly higher （P＜0.01）， and ultrasound irradiation could promote cellular uptake. NPs combined with ultrasound irradiation could inhibit the migration and invasion of M109 cells and block the cell cycle in the G0/G1 and G2/M phases. Compared with control group， the apoptosis rate of M109 cells and ROS level were increased significantly （P＜0.01）， while the MMP decreased significantly （P＜0.01） in the different concentration （100， 200， 300 μg/mL） groups of M109 cells. Compared with the mice in non-ultrasound group， the fluorescence intensity and tumor-targeting index of the tumor site in the 0 h ultrasound group were significantly enhanced （P＜0.05 or P＜0.01）. CONCLUSIONS NPs combined with ultrasound irradiation have a strong targeting effect on M109 cells in vitro and in vivo， the anti-tumor mechanism includes inhibiting cell migration and invasion， blocking cell cycle， and inducing apoptosis.

BACKGROUND:Traditional methods of urinary tract reconstruction are limited by donor scarcity,high complication rates,and suboptimal functional recovery.Tissue engineering strategies offer new directions in this field.Since the urinary tract is mainly composed of muscle tissue,the key is to find suitable seed cells and efficiently induce them to differentiate into smooth muscle cells.Comparative studies on the efficacy of different smooth muscle cell induction regimens are still lacking. OBJECTIVE:To isolate,culture,and identify human urine-derived stem cells,and to compare the effects of two different induction protocols. METHODS:Human urine-derived stem cells were isolated from urine samples of 11 healthy adult volunteers by multiple centrifugations.Surface markers were identified by flow cytometry.The multi-directional differentiation potential of human urine-derived stem cells was verified through osteogenic and adipogenic differentiation.Differentiation was induced by transforming growth factor-β1 or transforming growth factor-β1 combined with platelet derived growth factor for 14 days.Immunofluorescence staining and western blot assay were employed to compare the expression differences of smooth muscle-specific proteins(α-SMA and SM22). RESULTS AND CONCLUSION:(1)Urine-derived stem cells were successfully isolated from the eight urine samples of healthy people.These cells exhibit a"rice grain"-like morphology and possess a robust proliferative capacity.(2)Urine-derived stem cells exhibited high expression of mesenchymal stem cell surface markers(CD73,CD90,and CD44)and extremely low expression of hematopoietic stem cell surface markers(CD34 and CD45).These cells did not express CD19,CD105,and HLA-DR.(3)After osteogenic and adipogenic differentiation,the formation of calcium nodules and lipid droplets was observed,with positive staining results from Alizarin Red S and Oil Red O staining.(4)After 14 days of smooth muscle induction culture,immunofluorescence staining revealed that the smooth muscle differentiation rate of urine-derived stem cells treated with a combination of transforming growth factor-β1 and platelet derived growth factor was significantly higher compared to those treated with transforming growth factor-β1 alone(P<0.005).(5)After 14 days of smooth muscle induction culture,western blot assay further demonstrated that the expression levels of α-SMA and SM22 in the transforming growth factor-β1/platelet derived growth factor group were significantly elevated compared to those in the transforming growth factor-β1 only group(P<0.005).These findings confirm that urine-derived stem cells can be non-invasively isolated using multiple rounds of centrifugation.Compared with transforming growth factor-β1 alone,the combination of transforming growth factor-β1 and platelet derived growth factor can improve the efficiency of inducing urine-derived stem cells to differentiate into smooth muscle cells.

Objective:To evaluate the clinical efficacy of a novel reduction device in the treatment of A3N0/1 thoracolumbar fracture using navigation-assisted techniques.Methods:A retrospective analysis was conducted on 45 patients (29 males, 16 females; mean age 40.67±16.11 years, range 24-57) with thoracolumbar fractures who underwent fracture reduction and pedicle screw fixation via the Wiltse approach at Zhengzhou Orthopaedic Hospital between January 2022 and January 2023. Injury levels included: T 10 in 2 cases, T 11 in 5 cases, T 12 in 13 cases, L 1 in 20 cases, L 2 in 3 cases, L 3 in 2 cases. All patients underwent fracture reduction via the Wiltse approach using the spinal fracture reduction instrument for vertebral body reduction. Among them, 20 patients received O-arm navigation-assisted internal fixation and vertebral reduction (O-arm group), while 25 received C-arm fluoroscopy-guided internal fixation and vertebral reduction (C-arm group). Operative time, intraoperative blood loss, vertebral reduction time using the instrument, first-time screw placement success rate, screw placement accuracy, and complications were compared. Mid-vertebral body height ratio (MVBHr), local Cobb angle of the fractured vertebra, visual analogue scale (VAS) score, and Oswestry disability index (ODI) were compared preoperatively, at 1 week postoperatively, 3 months postoperatively, and final follow-up. Results:All surgeries were successfully completed in both groups. Operative time was significantly shorter in the O-arm group (106.8±14.4 min) than in the C-arm group (119.1±16.4 min, P<0.05). All patients were followed up for a mean duration of 15.9±3.9 months (range 12-20 months). Vertebral reduction time was significantly shorter in the O-arm group (11.0±2.2 min) than in the C-arm group (20.4±5.7 min, P<0.05). The first-time screw placement success rate was significantly higher in the O-arm group (100%) than in the C-arm group (95.3%, P<0.05). Screw placement accuracy (Grade I) was significantly higher in the O-arm group (117 screws, 97.5%) than in the C-arm group (136 screws, 90.7%, P<0.05). No cases of wrong-level surgery, infection, or spinal cord/nerve injury occurred. Both groups showed significant improvements in MVBHr, Cobb angle, VAS, and ODI at all postoperative time points compared to preoperative values ( P<0.05). At final follow-up, the O-arm group demonstrated significantly better outcomes than the C-arm group in MVBHr (90.6%±4.5% vs. 86.4%±6.9%, P<0.05), Cobb angle (7.6°±1.8° vs. 10.1°±3.2°, P<0.05), VAS (1.3±0.4 vs. 1.7±0.6, P<0.05), and ODI (4.6%±1.9% vs. 7.7%±2.0%, P<0.01). Conclusion:O-arm navigation-assisted intrasegmental push reduction for A3N0/1 type thoracolumbar fractures demonstrates advantages including faster and more accurate screw placement, precise reduction with improved outcomes, and significant postoperative pain relief.

AIM To study the chemical constituents from the sticks and leaves of Croton cascarilloides Raeusch.and their biological activities.METHODS The 95％ethanol extract from the sticks and leaves of C.cascarilloides was isolated and purified by MCI,silica gel,Sephadex LH-20 and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.LPS-induced NO RAW264.7 cell model induced by LPS was used to evaluate its anti-inflammatory activity in vitro.GES-1 injury model induced by taurocholic acid was used to screen the gastric mucosal protection activity.RESULTS Fourteen compounds were isolated and identified as bullatantriol(1),(-)-boscialin(2),(+)-dehydrovomifoliol(3),3-(hydroxylacetyl)-indole(4),pinoresinol(5),3,7-dimethyl-octa-1,7-diene-3,6-ol(6),(+)-syringaresinol(7),curcasinlignan B(8),cleomiscosin C(9),cleomiscosinD(10),2,6-dimethyl-octa-1,7-dien-3,6-diol(11),vanillin(12),vanillic acid(13),methyl vanillate(14).Compound 4 had certain anti-inflammatory activity,with IC50 values of 73.62 μmol/L.The protective rates of 25 μmol/L compounds 1-4,6,9-12 and 14 on gastric mucosal epithelial cells were 30.07％,34.18％,23.91％,30.92％,17.51％,19.69％,31.76％,22.46％,30.56％and 14.49％,respectively.CONCLUSION Compounds 1-14 are isolated from this plant for the first time.Compound 4 shows anti-inflammatory activity,1-4,6,9-12 and 14 show different degrees of gastric mucosal epithelial cell protective activity.

Objective To explore the effects of Shenyuan Yiqi Huoxue Capsules on cardiopulmonary exercise function and quality of life in patients with ischemic cardiomyopathy.Methods Totally 66 patients with ischemic cardiomyopathy were divided into treatment group and control group using random number table method,with 33 cases in each group.Both groups received standardized Western medicine treatment,and the treatment group was additionally given Shenyuan Yiqi Huoxue Capsules,the control group was given Shenyuan Yiqi Huoxue Capsules simulator,three capsules per time,three times a day,orally.The observation period was 30 days.To evaluate the efficacy of TCM syndrome in two groups.The observation indicators included baseline data,TCM syndrome scores,cardiopulmonary exercise test detection indicators,MLHFQ score,SF-36 score,echocardiographic indicators,N-terminal pro-B-type natriuretic peptide(NT-pro BNP)and safety indicators.Results The total effective rate of the treatment group was 93.94%(31/33),while that of the control group was 78.79%(26/33).The treatment group was significantly better than the control group(P<0.05).Compared with before treatment,the TCM syndrome scores of the two groups of patients decreased(P<0.05),and after treatment,the TCM syndrome score of the treatment group was lower than that of the control group(P<0.05).Compared with before treatment,the peak oxygen uptake(peak VO2),anaerobic threshold(AT)and peak oxygen pulse(peak O2 pulse)of both groups of patients increased,while the carbon dioxide ventilation equivalent slope(VE/VCO2 slope)decreased(P<0.05).After treatment,the treatment group had higher peak VO2,AT and peak O2 pulse than the control group,while the VE/VCO2 slope was lower than the control group(P<0.05).Compared with before treatment,both groups of patients showed a decrease in MLHFQ score and an increase in SF-36 score(P<0.05);after treatment,the MLHFQ score in the treatment group was lower than that in the control group,and the SF-36 score was higher than that in the control group(P<0.05).Compared with before treatment,both groups of patients showed a decrease in left ventricular end systolic diameter,left ventricular end diastolic diameter and NT-pro BNP levels,while myocardial motion index and left ventricular ejection fraction increased(P<0.05);after treatment,the treatment group showed a decrease in left ventricular end systolic diameter,left ventricular end diastolic diameter and NT-pro BNP levels,while myocardial motion index and left ventricular ejection fraction increased(P<0.05).During the treatment period,no adverse reactions such as dizziness,nausea,vomiting or diarrhea were found in both groups of patients.Conclusion Shenyuan Yiqi Huoxue Capsules can significantly improve the clinical efficacy of ischemic cardiomyopathy patients,alleviate TCM symptoms,improve their cardiorespiratory function and quality of life.

Objective:To compare the clinical efficacy of O-arm navigation-assisted percutaneous vertebroplasty (PVP) versus C-arm-guided PVP in the treatment of postoperative recurrent vertebral fractures following Kümmell′s disease.Methods:A retrospective cohort study was conducted to analyze the clinical data of 48 patients with postoperative recurrent vertebral fractures following Kümmell′s disease who were admitted to Zhengzhou Orthopedic Hospital from January 2021 to September 2024, including 16 males and 32 females, aged 51-85 years [(69.8±6.6)years]. Among them, 21 patients had stage I Kümmell′s disease and 27 stage II. Fractured vertebrae involved T 8-T 10 in 4 patients, T 11-L 2 in 29, and L 3-L 5 in 15. Twenty-five patients underwent O-arm navigation-assisted PVP (O-arm-assisted group) and 23 underwent C-arm-guided PVP (C-arm-guided group). The two groups were compared in terms of the operative time, intraoperative blood loss, bone cement volume, and bone cement filling saturation rate in the injured vertebral body. The visual analogue scale (VAS) scores and Oswestry disability index (ODI) values were also compared before operation, at 1 day, 1 month, 6 months after operation, and at the last follow-up. The excellent-good rate based on the MacNab criteria at the last follow-up and incidence of postoperative complications were detected. Results:All the patients were followed up for 6-24 months [(13.3±3.5)months]. There were no significant differences in the operative time, operative blood loss or bone cement volume between the two groups ( P>0.05). The O-arm-assisted group demonstrated a bone cement filling saturation rate of 96% (24/25) in the fractured vertebrae, significantly higher than 65% (15/23) in the C-arm-guided group ( P<0.05). The VAS scores before operation, at 1 day, and 1 month after operation were (8.4±1.0)points, (1.9±0.7)points, and (1.8±0.6)points, respectively in the O-arm-assisted group, while they were (8.3±0.8)points, (2.0±0.6)points, and (1.9±0.5)points, respectively in the C-arm-guided group ( P>0.05). The ODI values before operation, at 1 day, and 1 month after operation were 76.6±8.2, 20.4±4.5, and 19.8±4.1, respectively in the O-arm-assisted group, and 74.9±9.1, 21.3±3.6, and 20.9±3.2, respectively in the O-arm-assisted group ( P>0.05). At 6 months after operation and at the last follow-up, the VAS scores were (1.4±0.5)points and (1.5±0.5)points in the O-arm-assisted group, with significant improvement compared to (1.8±0.4)points and (1.9±0.3)points in the C-arm-guided group ( P<0.01); the ODI values were 17.8±3.2 and 18.2±3.5 in the O-arm-assisted group, with significant improvement compared to 19.9±3.1 and 21.3±4.0 in the C-arm-guided group ( P<0.05). Both groups demonstrated significant improvements in VAS scores and ODI values at 1 day, 1 month, 6 months after operation, and at the last follow-up, compared to those preoperatively ( P<0.05), while no statistically significant differences were found in VAS scores or ODI values at any postoperative timepoints ( P>0.05). According to the MacNab criteria, the O-arm-assisted group had a 100% (25/25) excellent-good rate, compared to 74% (17/23) in the C-arm-guided group ( P<0.05). The complication rate was 4% (1/25) in the O-arm-assisted group, significantly lower than 35% (8/23) in the C-arm-guided group ( P<0.05). Conclusion:O-arm navigation-assisted PVP for postoperative recurrent vertebral fractures following Kümmell′s disease offers advantages in precise cement delivery with sufficient dispersion, enhanced pain relief, functional recovery, improved quality of life, and reduced complication rates when compared to C-arm navigation-assisted PVP.

Objective To explore the clinical efficacy of dual plasma molecular adsorption(DPMAS)sequential plasma exchange(PE)artificial liver mode in the treatment of liver failure(LF).Methods Eighty-five patients with LF receiving the artificial liver treatment in the Affiliated Hospital of Zunyi Medical Univer-sity from January 2020 to December 2023 were selected as the study subjects and divided into the study group(n=52)and the control group(n=33)according to the different treatment modes.The study group conduc-ted DPMAS sequential PE treatment and the control group underwent the PE treatment.The liver function[total bilirubin(TBIL),alanine aminotransferase(ALT),aspartate aminotransferase(AST),serum albumin(ALB),globulin(GLO),prealbumin(PAB)],Hb,coagulation function[platelet(PLT),plasminogen activity(PTA),international normalized ratio(INR),fibrinogen(FIB)]before treatment and at 24 h after treatment were compared between the two groups.Results Compared with before treatment,the levels of TBIL,ALT,AST,GLO and Hb after the first and second treatment in the two groups were decreased,ALB level in the control group and PAB level after the second time treatment was increased(P＜0.05).Compared with after the first treatment,the levels of TBIL,ALT and GLO after the second treatment in the two groups and the levels of AST and Hb in the study group were decreased,ALB level in the study group and PAB level in the two groups were increased(P＜0.05).Compared with before treatment,the levels of PLT and FIB after the first treatment in the two groups and INR level in the control group were decreased,PTA level in the control group was increased(P＜0.05).Compared with before treatment,the levels of PLT,INR and FIB after the second treatment in the two groups were decreased,PTA level was increased(P＜0.05).Compared with be-fore treatment,the levels of PLT,INR and FIB after the second treatment in the two groups were decreased,and PTA level was increased(P＜0.05).Compared with after the first treatment,PTA level after the second treatment in the study group was increased and INR level was decreased.Conclusion PE and DPMAS sequen-tial PE all could improve the liver function in the patients with LF,moreover the two times treatment has more significant effect.

Objective This study aims to systematically evaluate the evidence-based medicine of traditional Chinese medicine(TCM)in the treatment of dry eye,analyze its efficacy and differences compared with modern Western medicine treatments,and provide a scientific basis for clinical application.Methods By searching Chinese and English databases(including CNKI,Wanfang Data,PubMed,Cochrane Library,etc.),we included meta-analyses and systematic reviews of TCM treatments for dry eye.Literature was screened according to strict inclusion and exclusion criteria,and relevant data were extracted and integrated for analysis.The Mantel-Haenszel fixed-effects model was used to calculate the relative risk(RR)and mean difference(WMD),with the effect size expressed as a 95％confidence interval,to analyze the efficacy in-dicators of TCM treatment for dry eye.Results A total of 21 articles were finally included,involving various TCM inter-ventions for dry eye(such as Chinese herbal medicine,acupuncture,and TCM external therapies).The results showed that TCM treatment was superior to Western medicine alone in improving the overall clinical efficacy[RR=1.63,95％CI(1.46,1.81),P＜0.05],prolonging tear film break-up time[MD=2.23,95％CI(1.85,2.60),P＜0.05],and increasing tear secretion[MD=1.49,95％CI(1.04,1.94),P＜0.05].In addition,acupuncture,the combination of Chinese herbal medicine and Western medicine,and TCM external therapies all demonstrated unique advantages in improving dry eye symptoms and tear function.Conclusion TCM treatment for dry eye shows significant efficacy in key indicators such as overall clinical efficacy,tear film break-up time,and tear secretion,with a high level of safety.