1.Preparation and in vitro evaluation of platelet membrane biomimetic liposomes loaded with vincristine sulfate
Jing XIAO ; Xunyi YOU ; Along ZHANG ; Rui ZHONG ; Jiaxin LIU ; Ye CAO ; Hong WANG
Chinese Journal of Blood Transfusion 2025;38(5):652-659
Objective: To prepare platelet membrane biomimetic liposomes loaded with vincristine sulfate (VCR) for targeted delivery to tumor. Methods: Vincristine sulfate liposomes (LIPO) were prepared using the pH-gradient method, followed by the fusion of platelet membranes and subsequent drug loading to obtain platelet membrane biomimetic liposomes (PLM-LIPO). The particle size, polydispersity index (PDI), Zeta potential, and drug encapsulation efficiency (EE%) of both liposomes were characterized. The tumor-targeting capability was evaluated through in vitro cellular experiments and in vivo biodistribution studies. Results: The optimal preparation conditions for LIPO were determined as follows: DPPC-to-cholesterol molar ratio of 1∶1, internal aqueous phase of 0.3 M pH 4.0 citrate buffer, external aqueous phase of 1 M Na
HPO
solution, drug-to-lipid ratio of 1∶10, drug loading temperature of 60℃, and loading time of 10 minutes. The LIPO exhibited a mean particle size of (147.3±2.24) nm, PDI of 0.078±0.014, Zeta potential of (-3.54±0.75) mV, and EE% of 91.37±0.47. For PLM-LIPO, prepared via membrane fusion followed by drug loading, the mean particle size was (185.3±3.61) nm, PDI was 0.075±0.022, Zeta potential was (-18.91±1.54) mV, and EE% was 63.36±2.45. In the CD62P validation experiment, the fluorescence intensity of PLM-LIPO was five times higher than that of LIPO. In vitro cellular uptake experiments revealed that PLM-LIPO showed 1.3-fold and 1.2-fold higher uptake rates compared to LIPO at 6 h and 12 h, respectively. In vivo experiments demonstrated that 1h after administration, the accumulation of PLM-LIPO at tumor sites was 4-fold higher than that of LIPO and 6-7 times higher than that in healthy mice. Conclusion: The platelet membrane biomimetic liposomes loaded with vincristine sulfate were successfully developed. Both cellular uptake and tissue distribution studies confirmed the PLM-LIPO enhanced tumor-targeting capability.
2.Design, synthesis and anti-Alzheimer's disease activity evaluation of cinnamyl triazole compounds
Wen-ju LEI ; Zhong-di CAI ; Lin-jie TAN ; Mi-min LIU ; Li ZENG ; Ting SUN ; Hong YI ; Rui LIU ; Zhuo-rong LI
Acta Pharmaceutica Sinica 2025;60(1):150-163
19 cinnamamide/ester-triazole compounds were designed, synthesized and evaluated for their anti-Alzheimer's disease (AD) activity. Among them, compound
3.Construction of Multidimensional Evaluation System for Health Status in Modern China's Ideal Life
Rui XU ; Han ZHOU ; Zhong WANG ; Yongyan WANG
Journal of Traditional Chinese Medicine 2025;66(7):657-662
The evolving health demands of contemporary society urgently call for adaptive adjustments in health policies and a comprehensive enhancement of perspectives, horizons, and viewpoints. Grounded in the solid foundation of traditional Chinese culture, this paper proposed a multidimensional evaluation system for assessing the health status of modern China's ideal life. Drawing upon the philosophical insights of Confucianism, Daoism, Mohism, Song-Ming Neo-Confucianism, and their subsequent developments, as well as core cultural values such as harmony (和), happiness (福), and joy (乐), this system established a fundamental framework for evaluating health in modern ideal life. The framework consists of eight indices, i.e. harmony index, diligence index, peace of mind index, happiness index, compassion index, innovation index, fulfillment index, and adaptability index. The purpose of constructing this system is to comprehensively and deeply reflect the physical and mental characteristics and behavioral patterns of the Chinese population, and to provide a holistic evaluation framework for advancing the foundation of health in modern ideal life, offering solid scientific support for formulating health policies that align with China's national context.
4.Effect of Shenmai injection on the preservation quality of suspended red blood cells prepared from high hemoglobin population in Xizang plateau
Rui ZHONG ; Zeng HE ; Hong WANG ; Wanjing LI ; Jiaxin LIU ; Ye CAO
Chinese Journal of Blood Transfusion 2025;38(3):327-332
[Objective] To investigate the effect of Shenmai injection on the preservation quality of suspended red blood cells of high hemoglobin population in Xizang plateau. [Methods] Whole blood (400 mL, n=8) collected by the Xizang Autonomous Region Blood Center was centrifuged at 3 000 g for 10 minutes to remove most of the plasma, followed by the addition of 100 mL of MAP preservation solution to obtain plateau suspended red blood cells, which were then divided into three equal portions. One portion was the control group, and another part had 15 mL of MAP preservation solution added, which was the dosage group. The third portion involved diluting Shenmai injection with MAP, followed by addition of 15 mL of MAP preservation solution containing Shenmai to the red blood cells, resulting in a final concentration of Shenmai injection of 1%, which was the Shenmai group. Blood routine, pH value, electrolytes, glucose, lactate, free Hb, adenosine triphosphate (ATP), P50, phosphatidylserine (PS) and other indicators were detected at day 1, 21 and 35, respectively. [Results] The Hb concentration and Hct of the dosage group and the Shenmai group were significantly lower than those of the control group, with values of (179.3±17.8) vs (181.0±17.1) vs (199.1±19.5) g/L for Hb concentration and (53.2±2.6)% vs (53.3±2.5)% vs (58.4±3.1)% for Hct. The three groups maintained this pattern until the end of storage. In the middle and late stages of preservation, the glucose and Na+ contents in the dosage group and the Shenmai group were higher than those in the control group, while the lactate and K+ contents were lower than those in the control group. At the end of storage, the glucose and Na+ content of the Shenmai group was higher than that of the dosage group, while the lactate and K+ content were lower than that of the dosage group. From day 1 to day 35 of storage, the hemolysis rate of the Shenmai group was significantly lower than that of the control group and the dosage group. On day 21 and 35 of storage, the PS expression rate in the Shenmai group was significantly lower than that in the control group and the dosage group, which were (6.52±0.40)% vs (7.24±0.91)% vs (8.27±0.93)% and (7.29±0.53)% vs (9.37±0.82)% vs (8.39±0.76)%, respectively. [Conclusion] The hemolysis rate and PS of suspended red blood cells of Xizang high altitude prepared by adding Shenmai injection were significantly lower than those in the control group and the dosage group, which was conducive to reducing hemolysis and slowing down the aging of red blood cells, and had a certain improvement on the preservation quality of suspended red blood cells in Xizang plateau people.
5.Study on soluble microneedles loaded with aPexos and EGCG for healing of burn wounds in rats
Along ZHANG ; Wanjing LI ; Hong WANG ; Jiaxin LIU ; Ye CAO ; Rui ZHONG
Chinese Journal of Blood Transfusion 2025;38(3):333-342
[Objective] To use Chitosan Methacryloyl (CSMA) loaded with artificial platelet-derived exosomes (aPexos) as the needle body material, and gelatin and carboxymethyl chitosan (CMCS) loaded with Epigallocatechin gallate(EGCG) as the backing material to prepare microneedles, aims to investigate the therapeutic effect and mechanism of aPexos-EGCG microneedles in the treatment of burn wound healing. [Methods] First, aPexos were extracted using ultrasound and gradient ultracentrifugation. The concentration, morphology, and growth factor content (TGF-β1, PDGF-BB, VEGF) of the exosomes were assessed using NTA, transmission electron microscopy, and ELISA kits. The aPexos and EGCG were then dissolved in the needle body and backing materials, respectively. The aPexos and EGCG were loaded into the needle body and backing layer, respectively. The morphology, mechanical properties, and puncture performance of the microneedles were examined, and the preparation conditions for the microneedles loaded with aPexos-EGCG were optimized. Finally, animal experiments and tissue staining were conducted to assess the efficacy of the aPexos-EGCG microneedles in promoting burn wound healing in rats. [Results] The size distribution of aPexos was mainly in the range of 50-150 nm, with an average diameter of 132.7±3.8 nm and an average concentration of approximately 2.88×1013±3.62×1012 particles/mL. The concentrations of TGF-β1, PDGF-BB, and VEGF were approximately 1 363±135.9 ng/mL, 63.5±14.8 ng/mL, and 1 606.0±77.5 pg/mL, respectively. Mechanical property testing of the microneedles indicated that, compared with the blank microneedles, the drug-loaded microneedles had sufficient piercing ability to penetrate the stratum corneum of the skin. The results of the rat burn wound healing experiment showed that the wound healing rates of the d 3, d 7, aPexos-EGCG microneedle group compared to the control group showed significant differences, with respective values of (47.64±12.5)% vs (18.11±6.40)%, (87.45±5.57)% vs (79.85±5.03)%(P<0.05). Hematoxylin-eosin (HE) staining revealed more intact epidermal layers in the aPexos-EGCG microneedle group. Masson staining showed that the collagen deposition percentage in the aPexos-EGCG microneedle group was higher than in the control group. Immunofluorescence staining results indicated that the aPexos-EGCG microneedle group had a reduced content of M1-type macrophages and an increased content of M2-type macrophages. Additionally, the vascular markers CD31 and α-SMA showed elevated expression in this group, with significant differences compared to the other groups (P<0.05). [Conclusion] The aPexos-EGCG microneedle developed in this study exhibits good mechanical properties, capable of successfully penetrating the skin's stratum corneum and delivering aPexos and EGCG. aPexos promotes wound repair, while EGCG regulates the ratio of M1 and M2 macrophages at the wound site and suppresses the inflammatory response. This microneedle can effectively promote wound healing in rats with burn injuries, offering a novel approach for tissue repair.
6.Effect Mechanism and Law of Sterilization by 60Co-γ Ray Irradiation on Chemical Composition of Chinese Materia Medica: A Review
Tingting ZHU ; Jian RANG ; Rangyanpo LUO ; Rui GU ; Yue YANG ; Si LU ; Shihong ZHONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(18):306-314
60Co-γ ray irradiation has the unique advantages of high efficiency, strong penetration, operation at room temperature and no residue, which has been widely used in the sterilization of Chinese medicinal materials, decoction pieces, Chinese patent medicine. However, the irradiation effect may cause changes in the content of chemical components in Chinese materia medica or the emergence of new radiolysis products, leading to reduced efficacy and uncontrollable safety risks. This paper reviewed the relevant literature at home and abroad, summarized the effect of irradiation sterilization on various types of chemical compositions of Chinese medicinal materials and their preparations, and analyzed and explored the rule of change. The results showed that the content changes of various chemical components in Chinese materia medica after 60Co-γ ray irradiation sterilization varied. The contents of most flavonoids, terpenoids, phenylpropanoids and quinones decreased after irradiation, and the degree of decrease increased with the elevated irradiation dose. The contents of lignans, alkaloids, isoflavones and some terpenoids did not change significantly before and after irradiation, while the content changes of triterpenoid saponins, dihydroflavonols, chalcones, sugars and glycosides after irradiation were not yet uniform. Therefore, it is recommended to pay attention to the compositional changes of irradiated Chinese medicines, strengthen the research on the standards of irradiated Chinese medicines, and standardize the irradiation and sterilization of Chinese medicines in order to promote the healthy and rational application of irradiated Chinese medicines.
7.Comparison of Protein and Polypeptide Components and Antithrombotic Activity In Vitro of Three Preparations Containing Hirudo
Wanling ZHONG ; Yunnan MA ; Jinhong YE ; Xiaoyu FAN ; Huijuan SHEN ; Rui YUAN ; Yaxuan ZHANG ; Zhuyuan LIU ; Shouying DU ; Pengyue LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):184-194
ObjectiveTo compare the contents and relative molecular weight distributions of proteins and polypeptides in Naoxuekang dropping pills, Huoxue Tongmai capsules and Maixuekang capsules of Hirudo single medicinal preparations, to evaluate the in vitro anticoagulant, antiplatelet and fibrinolytic activities of the three preparations, and to investigate the effects of temperature, pH and digestive enzymes on the anticoagulant activities of the three preparations. MethodsThe contents of soluble proteins and polypeptides in the three preparations were determined by bicinchoninic acid assay(BCA) and Bradford method, and the relative molecular weight distributions of the three preparations were determined by electrophoresis combined with gel chromatography. The antithrombin activity of the three preparations was evaluated by fibrinogen-thrombin time(Fibg-TT) method, and their anticoagulant activities were further assessed by the elongations of activated partial thromboplastin time(APTT), prothrombin time(PT) and thrombin time(TT). The antiplatelet aggregation activities of the three preparations were measured by turbidimetry and the fibrinolytic activities were measured by fibrin plate method. Relative TT was used as index to investigate the effects of temperature, pH and digestive enzyme buffer on anticoagulant activities of the three preparations. ResultsAt the lowest single dosage, the contents of proteins and polypeptides were in the order of Maixuekang capsules>Huoxue Tongmai capsules>Naoxuekang dropping pills. Both Huoxue Tongmai capsules and Maixuekang capsules had 11 electrophoretic bands between 4.0 kDa and 90 kDa, the bands of Maixuekang capsules were more clear in the range of >25 kDa, and there was 1 obvious band at 14 kDa for the two capsules. Huoxue Tongmai capsules had one specific band at 9.0 kDa and Maixuekang capsules had one specific band at 48.0 kDa. Naoxuekang dropping pills only had 2 electrophoretic bands at 6.5 kDa and 8.5 kDa, primarily containing peptides below 2 kDa, most of which were oligopeptides. The anticoagulant activity concentrations of the three preparations exhibited a certain dose-dependent effect. At the lowest single dosage, The anticoagulant activity concentrations were ranked as Naoxuekang dropping pills>Huoxue Tongmai capsules>Maixuekang capsules. The prolongation effect of the three preparations on coagulation time was dose-dependent. At the same concentration, the prolongation effect of Naoxuekang dropping pills and Huoxue Tongmai capsules was APTT prolongation rate>TT prolongation rate>PT prolongation rate, whereas for Maixuekang capsules, the sequence was TT prolongation rate>APTT prolongation rate>PT lengthening rate. At the single minimum dosage, the order of APTT prolongation rate was Maixuekang capsules>Huoxue Tongmai capsules≈Naoxuekang dropping pills, the order of PT prolongation rate was Naoxuekang dropping pills≈Maixuekang capsules>Huoxue Tongmai capsules, and the order of TT prolongation rate was Maixuekang capsules>Huoxue Tongmai capsules>Naoxuekang dropping pills. The three preparations showed dose-dependent effects on platelet aggregation induced by adenosine diphosphate(ADP) and arachidonic acid(AA), and the effect induced by ADP was stronger than that induced by AA. The anti-platelet aggregation effect of Naoxuekang dropping pills was significantly stronger than that of Maixuekang capsules(P<0.01), whereas Huoxue Tongmai capsules had the effect of promoting platelet aggregation. None of the three preparations had the ability to dissolve fibrin. The anticoagulant activity of Naoxuekang dropping pills was least affected by heating, while the activities of the two capsules decreased significantly within 5 min above 80 ℃, and continued to decrease within 2 h. Compared with pure water, the anticoagulant activities of the three preparations could be increased by 1-3 times under strong acidity(pH 1-3). In the pepsin buffer, the anticoagulant activity of Naoxuekang dropping pills could be increased by 1-3 times, while the anticoagulant activities of Huoxue Tongmai capsules and Maxuekang capsules were significantly decreased, the lowest levels were about 60% and 20%, respectively. In trypsin buffer, the anticoagulant activities of Naoxuekang dropping pills, Huoxue Tongmai capsules and Maixuekang capsules decreased significantly, and the lowest levels decreased to about 41%, 41% and 35%, respectively. ConclusionThe contents of proteins and polypeptides and relative molecular weights of the preparations derived from lyophilized fresh Hirudo powder, dried Hirudo powder and reflux extract of Hirudo decrease sequentially, and the anticoagulant activity decrease gradually, but the anticoagulant pathway is different. And the anti-platelet aggregation activity of the reflux extract is significantly enhanced. The heat resistance and gastrointestinal stability of the three preparations increase successively, and the first two are suitable for enteric-soluble preparations, while the latter is suitable for routine oral administration. The above results can provide data reference for the rationality of different preparation methods, active substances, pharmacodynamics and mechanism of Hirudo preparations.
8.Investigation of tick species and potential pathogenic ricks in certain areas of Wuwei City,Gansu Province
Rui-Shan LI ; Zhen HE ; Xiang YUAN ; Shi-Wei SUN ; Yi-Wen LIU ; Wen-Kai ZHANG ; Lin ZHANG ; Yu-Hua WANG ; Zhen-Hua LU ; Zhao-Hua JI ; Zhong-Jun SHAO
Chinese Journal of Zoonoses 2024;40(4):328-333
To understand the distribution of ticks in the Wuwei Region,enrich tick species data,and provide a basis for the prevention of tick-borne diseases,tick were collected using flagging and tick-picking methods during the highest activity period from April to September in 2021 and 2022 in the mountainous areas of Wuwei City.The ticks were identified based on morpho-logical and molecular biological characteristics,and characteristic sequences were obtained.A systematic evolutionary tree was constructed using the neighbor-joining method in MEGA 11.0 software.In total,7 342 ticks collected in Wuwei,which be-longed to 5 species from 4 genera with in the Ixodidae family,which included Dermacentor nuttalli,Hyalomma asiaticum,Ixodes canisuga,Haemaphysalis longicornis and Haema-physalis danieli.Ticks of the same species clustered together into the same branch of an evolutionary tree.In the Wuwei Re-gion,five common tick species are found across various habi-tats,with each habitat featuring different distributions of tick species and populations.The Dermacentor nuttalli is the dom-inant tick species in this area.
9.Optimized expression of the diphtheria toxin mutant CRM197 in Escherichia coli and population analysis of serum antibody levels
Xiao-Li CHEN ; Yi-Xin GU ; Hai-Rui WANG ; Gui-Lan ZHOU ; Xin ZHANG ; Chang LIU ; Jian-Zhong ZHANG ; Zhu-Jun SHAO ; Mao-Jun ZHANG
Chinese Journal of Zoonoses 2024;40(5):430-434
A prokaryotic expression vector for the mutant diphtheria toxin CRM197 was constructed and expressed in Esch-erichia coli cells.Anti-CRM197 antibody concentrations were detected in serum samples of healthy volunteers.The crm 197 gene was codon-optimized in E.coli and cloned into the plasmid pET28a(+)under optimized expression conditions.CRM197 was purified using Ni-NTA spin columns and ion exchange chromatography,and confirmed by western blot analysis.The puri-fied CRM197 was used to detect specific anti-CRM197 antibody levels in serum samples of different age groups.The results showed that soluble codon-optimized CRM197 was successfully expressed under optimized expression conditions.The purity of CRM197 was more than 95%,as determined with Ni-NTA spin columns and ion exchange chromatography,consistent with the single specific bands obtained by western blot analysis and detection of serum levels of the anti-CRM197 antibody.Collec-tively,these results confirmed that the proposed expression strategy achieved high-yield production of soluble CRM197,al-though high levels in human serum may affect evaluation of immune interactions with glycan-CRM197 conjugates for applica-tion as a diagnostic antigen.The diphtheria mutant toxin CRM197 is used in many conjugate vaccines.The synthetic crm 197 gene with codon optimization in pET28a was transformed into E.coli Origami B(DE3)cells.CRM197 was induced by isopro-pyl β-d-1-thiogalactopyranoside and high level accumulation of soluble CRM197 was purified using Ni-NTA spin columns and ion exchange chromatography.The purity of the final prepara-tion reached 95%.CRM197 was used to detect the concentra-tions of the anti-CRM197 antibody in serum samples of healthy volunteers of different ages.The proposed expression strategy yielded high production of CRM197,which could interfere with evaluations of induced immune interactions by glycan-CRM197 conjugates and prohibit application as a diagnostic antigen.
10.Effect of Cichorium glandulosum Boiss.et Huet.on fecal bile acid profile in obese mice based on targeted metabolomics technique
Shuwen QI ; Yewei ZHONG ; Abudurexiti ADALAITI ; Rui ZHANG ; Wenhui HOU ; Chunzi ZHANG ; Xiaoyan MA ; Xiaoli MA
Acta Laboratorium Animalis Scientia Sinica 2024;32(9):1122-1138
Objective To investigate the effect of ethanolic extract of Cichorium glandulosum Boiss.et Huet.on fecal bile acid profiles in high-fat-diet-induced obese mice.Methods Twenty-four 6-week-old C57 BL/6 male mice were divided randomly into normal,model,drug-administration,and metformin groups.Mice in the normal group were fed a regular diet and mice in the other three groups were given high-fat diets.The drug-administration group was gavaged with 10 mL/kg ethanol extract of Cichorium glandulosum Boiss.et Huet.daily,and the metformin group was gavaged with 10 mL/kg metformin daily.After 10 weeks,livers were collected to measure hepatic total triglycerides(TG),total cholesterol(TC),low-density lipoprotein-cholesterol(LDL-C),and high-density lipoprotein(HDL)-C.Feces were collected and analyzed.Results Body weight(P<0.0001),liver TG(P<0.05),and TC(P>0.05)were all significantly higher in model mice compared with normal mice,while LDL-C(P>0.05)and HDL-C(P<0.001)were significantly lower,indicating abnormal weight gain and lipid metabolism.Alcoholic extract of Cichorium glandulosum Boiss.et Huet.significantly reduced body weight(P<0.0001),liver TG(P<0.0001),serum TG(P<0.05),TC(P<0.01),and LDL-C(P<0.05)in mice.Methodsological validation showed that the current method could accurately quantify 52 bile acids in feces.Analysis of the concentration of each type of bile acid revealed that alcoholic extract of Cichorium glandulosum Boiss.et Huet.significantly increased the secondary/primary bile acid ratio(P<0.05).Multivariate analysis showed that the bile acid metabolic pattern was significantly altered in all groups.Eight differential bile acids were screened in the drug-administration group relative to the model group using variable importance of projection>1 and P<0.05.A search of the Kyoto Encyclopedia of Genes and Genomes database revealed that the differential bile acids were mainly involved in the secondary bile acid biosynthesis pathway.Correlation analysis showed that four differential bile acids,deoxycholic acid(r,=0.6445,P<0.001),isolithocholic acid(r,=0.5879,P<0.01),3β-deoxycholic acid(r,=0.6649,P<0.001),and ω-rhamnoglutaric acid(rs=0.5387,P<0.01),in feces were strongly positively correlated with body weight.Conclusions Cichorium glandulosum Boiss.et Huet.alcoholic extract may play a role in weight reduction and amelioration of dyslipidemia by modulating secondary bile acid biosynthesis and altering fecal bile acid metabolic profiles.

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