Objective:To prepare a novel ginseng polypeptide thermosensitive hydrogel,and to investigate its repair effect on heat-induced skin injury in the rats and explore the underlying mechanisms.Methods:Thermosensitive hydrogels were formulated using Pluronic F127 and β-sodium glycerophosphate(β-GP),and their phase transition temperatures,spatial structures,elemental compositions,and water retention capacities were evaluated.The rat models of heat-induced skin injury were established and the model rats were divided into PBS group,Gel group,and ginseng polypeptide gel(GP-Gel)group.After 11 d of treatment,the morphological changes of wound and collagen deposition in the wound of the rats in various groups were observed by HE and Masson staining.Immunohistochemistry was used to detect the expression levels of α-smooth muscle actin(α-SMA),connective tissue growth factor(CTGF),basic fibroblast growth factor(bFGF),proliferating cell nuclear antigen(PCNA),cell proliferation marker Ki67,epidermal growth factor(EGF),CD31,vascular endothelial growth factor(VEGF),P50 and P65 proteins in the skin wound tissue of the rats in various groups.Western blotting method was used to detect the expression levels of Toll-like receptor 4(TLR4)in the skin wound tissue of the rats in various groups.ELISA method was used to measure the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-15(IL-15),and interleukin-10(IL-10)in the serum of the rats in various groups.Results:Compared with PBS and Gel groups,the wound area of the rats in GP-Gel group was reduced(P＜0.01),the expression levels of PCNA,Ki67,EGF,CD31,VEGF,α-SMA,and CTGF proteins in the skin wound tissue were increased(P＜0.05 or P＜0.01),and the expression levels of P65 and TLR4 proteins were decreased(P＜0.01);the level of anti-inflammatory factor IL-10 in serum was increased(P＜0.01),while the levels of pro-inflammatory factors TNF-α,IL-1β,IL-6 and IL-15 were decreased(P＜0.05 or P＜0.01).Conclusion:The ginseng polypeptide thermosensitive hydrogel promotes the repair of heat-induced skin injury by enhancing cell proliferation,collagen synthesis,angiogenesis,and reducing inflammatory responses.

Objective To establish an initial diagnostic method for β-thalassemia (BT) using a double antibody sandwich ELISA approach. Methods The hybridoma producing monoclonal antibodies against anti-HbF were screened using a trailing count method. The mAbs were evaluated through ELISA,modified immunocytochem-istry,and Western blot analysis. A double antibody-sandwich ELISA assay was established by labeling the pairs of mAbs with ALP using the glutaral method,and this detection system was used to analyze 40 serum samples. Results The results demonstrate the successful generation of nine hybridoma cell lines capable of secreting highly active anti-HbF monoclonal antibodies (mAbs). Specifically,four mAbs (3F7,4G1,6C1,and 9H7) exhibited exclusive reactivity towards HbF without any cross-reactivity with hemoglobin variants (HbA and HbA2). These four HbF-specific mAbs displayed exceptional specificity and sensitivity,with a maximum titer of 1:256000 and the highest affinity constant (Ka) recorded at 2.36×108 L/mol. Among these mAbs,optimal antibody pairing was achieved using capture antibody 3F7 in conjunction with ALP-4G1 for the development of a sandwich ELISA detec-tion method. By employing this approach,fetal and healthy human blood samples were successfully quantified for HbF levels with an impressive detection sensitivity reaching up to 80％. Conclusion This sandwich ELISA dem-onstrated precise quantification of HbF levels,making it suiTab.for both research and diagnostic purposes in the field of β-thalassemia.

Objective To observe the clinical efficacy of Ziwu Liuzhu(midnight-midday ebb and flow,the movement of qi and blood in the meridians following heavenly-stems and earthly-branches cycle)timing point application in treating pediatric pneumonic cough of phlegm-heat obstructing lung syndrome.Methods A total of 300 cases of children with pediatric pneumonic cough of phlegm-heat obstructing lung syndrome admitted to the First Clinical Hospital of Jilin Academy of Traditional Chinese Medicine from January 2023 to April 2024 were randomly divided into observation group and control group according to random number table method,150 cases in each group,and both groups of children were given the conventional symptomatic treatment with western medicine,while the control group was given Cefuroxime Sodium intravenous drip.The observation group was treated with Ziwu Liuzhu timing point application on the basis of treatment of the control group.After treatment,the clinical efficacy of the two groups was evaluated,and the changes in traditional Chinese medicine(TCM)syndrome scores before and after treatment were observed,and the symptoms such as disappearance time of lung rale,recovery time of body temperature,recovery time of heart rate,and return to normal time of X-ray findings of the two groups were observed.The changes of red blood cell count(RBC),white blood cell count(WBC),platelet count(PLT)and C-reactive protein(CRP)levels before and after treatment were compared between the two groups.The safety and occurrence of adverse reactions in the two groups were also evaluated.Results(1)The total effective rate was 94.67%(142/150)in the observation group and 84.67%(127/150)in the control group.The efficacy of the observation group was superior to that of the control group,the difference being statistically significant(P<0.05).(2)After treatment,the TCM syndrome scores of the children in the two groups were improved significantly(P<0.05),and the improvement in the observation group was significantly superior to that in the control group,the difference being statistically significant(P<0.05).(3)After treatment,the observation group was superior to the control group in the disappearance time of lung rales,recovery time of body temperature,recovery time of heart rate,and return to normal time of X-ray findings,the difference being statistically significant(P<0.05).(4)After treatment,the RBC,WBC,PLT and CRP levels of the children in the two groups were improved significantly(P<0.05),and the improvement in the observation group was significantly superior to that in the control group,the difference being statistically significant(P<0.05).(5)The incidence rate of adverse reactions in the observation group was 10.67%,and that in the control group was 16.00%;the intergroup comparison showed that the difference being not statistically significant(P>0.05).Conclusion Ziwu Liuzhu timing point application in treating pediatric pneumonic cough of phlegm-heat obstructing lung syndrome can effectively improve the children's clinical symptoms,promote their recovery of lung function,alleviate the inflammatory reaction,which is safety,and with less adverse reaction.

Objective To establish an initial diagnostic method for β-thalassemia (BT) using a double antibody sandwich ELISA approach. Methods The hybridoma producing monoclonal antibodies against anti-HbF were screened using a trailing count method. The mAbs were evaluated through ELISA,modified immunocytochem-istry,and Western blot analysis. A double antibody-sandwich ELISA assay was established by labeling the pairs of mAbs with ALP using the glutaral method,and this detection system was used to analyze 40 serum samples. Results The results demonstrate the successful generation of nine hybridoma cell lines capable of secreting highly active anti-HbF monoclonal antibodies (mAbs). Specifically,four mAbs (3F7,4G1,6C1,and 9H7) exhibited exclusive reactivity towards HbF without any cross-reactivity with hemoglobin variants (HbA and HbA2). These four HbF-specific mAbs displayed exceptional specificity and sensitivity,with a maximum titer of 1:256000 and the highest affinity constant (Ka) recorded at 2.36×108 L/mol. Among these mAbs,optimal antibody pairing was achieved using capture antibody 3F7 in conjunction with ALP-4G1 for the development of a sandwich ELISA detec-tion method. By employing this approach,fetal and healthy human blood samples were successfully quantified for HbF levels with an impressive detection sensitivity reaching up to 80％. Conclusion This sandwich ELISA dem-onstrated precise quantification of HbF levels,making it suiTab.for both research and diagnostic purposes in the field of β-thalassemia.

OBJECTIVE To prepare nanoparticle-based targeting preparation loaded with triptolide(TP),and evaluate its quality,targeting ability and cytotoxic effects.METHODS Polymer nanoparticles carrying TP-targeted folic acid(FA)receptor(TP@PLGA-PEG-FA)were fabricated using poly(lactic-co-glycolic acid)/polyethylene glycol/FA(PLGA-PEG-FA)as the carrier by emulsion and volatilization technique.The morphology and distribution were observed,and their particle size,Zeta potential,polydispersity index(PDI),drug loading capacity and encapsulation efficiency were measured.Their stability,blood compatibility,in vitro drug release,uptake by RAW264.7 cells(localization with fluorescent dye Cy3.5),and in vitro cytotoxicity were evaluated.RESULTS TP@PLGA-PEG-FA exhibited spherical shape and uniform distribution,with particle size of(122.60±0.02)nm,Zeta potential of(-17.6±0.6)mV,and PDI of 0.26±0.02;drug loading capacity and encapsulation efficiency of TP were measured to be(7.78±0.05)%and(68.62±0.03)%,respectively.The hemolysis rates of 100,200,300,400 μg/mL TP@PLGA-PEG-FA were 0.77%,0.92%,1.34%and 1.63%,respectively.There were no significant changes in particle size,PDI and Zeta potential when TP@PLGA-PEG-FA were placed in 4℃water for 14 days and in DMEM culture medium containing 10%fetal bovine serum at 37℃for 12 h.The cumulative release rate of TP@PLGA-PEG-FA was(84.83±0.29)%in phosphate buffer at pH5.5 for 72 h,which was significantly higher than the cumulative release rates in phosphate buffer solutions at pH7.4 and 6.5 for 72 h[(42.37±0.35)%and(63.83±0.29)%,respectively](P＜0.05).Activated RAW264.7 cells took up significantly more Cy3.5@PLGA-PEG-FA than they took up Cy3.5@PLGA-PEG-FA+free FA and Cy3.5@PLGA-PEG.When the mass concentration of TP was≥15.63 ng/mL,the survival rates of activated cells in the TP@PLGA-PEG-FA groups were significantly lower than those of the same mass concentration of free TP groups(P＜0.05).CONCLUSIONS The prepared TP@PLGA-PEG-FA has high stability,good blood compatibility,active targeting and cytotoxicity to inflammatory cells.

Objective:To discuss the effect of royal jelly acid(10-HDA)on the proliferation and migration of the human colon cancer SW620 cells based on the network pharmacology,and to clarify its related molecular mechanism.Methods:The active ingredients such as 10-HDA and their corresponding targets were retrieved by using the keyword"royal jelly"from the Traditiomal Chinese Medicine Systems Pharmacology(TMSCP)Database and the Traditiomal Chinese Medicine Integrated Database(TCMID);the small molecule targets were predicted by the Swiss Target Prediction Database.The GeneCards Database and the Online Mendelian Inheritance in Man(OMIM)Database were used to obtain the targets with the keyword"Colon Cancer";the protein-protein interaction(PPI)network was constructed by using the String Database and Cytoscape 3.8.0 Software to screen the core targets;the Gene Ontology(GO)function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis were analyzed by Metascape Database;the specific ingredient 10-HDA was screened for the in vitro activity experiments.The human colon cancer SW620 cells with good growth status were divided into control group and different doses(1,5,10,15,and 20 mmol·L-1)of 10-HDA groups.The viabilities of the cells in various groups were detected by MTT method and the survival rates of the cells were calculated.The SW620 cells were divided into control group,low dose(5 mmol·L-1)of 10-HDA group,middle dose(10 mmol·L-1)of 10-HDA group,and high dose(15 mmol·L-1)of 10-HDA group;Hoechst33342 staining method was used to observe the morphology of the cells in various groups;cell scratch test was used to detect the scratch healing rates of the cells in various groups;flow cytometry was used to detect the percentages of the cells at different cell cycles in various groups;biochemical method was used to detect the activities of total antioxidant capacity(T-AOC)and superoxide dismutase(SOD)in the cells in various groups;Western blotting method was used to detect the expression levels of B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),cysteine-containing aspartate proteolytic enzyme-3(Caspase-3),cysteine-containing aspartate proteolytic enzyme-9(Caspase-9),glycogen synthase kinase 3β(GSK3β),β-catenin,and cyclin D1 proteins in the cells in various groups.Results:Six active ingredients of royal jelly were screened out by the TCMSP Database,and 28 core targets of 10-HDA in the treatment of colon cancer were obtained.The GO function enrichment analysis mainly included the signaling pathways such as cell proliferation and apoptosis.The KEGG signaling pathway enrichment analysis included the cell cycle,prostate cancer,cell senescence,and p53 signaling pathways;the GSK3β/β-catenin signaling pathway was closely related to the cell cycle.Compared with control group,the viabilities of the cells in 5,10,15,and 20 mmol·L-110-HDA groups were decreased in a dose-dependent manner(P<0.05 or P<0.01),the numbers of apoptotic cells in different doses of 10-HDA groups were significantly increased,and the scratch healing rates of the cells were significantly decreased(P<0.05 or P<0.01);the percentages of the cells at S phase in middle and high doses of 10-HDA groups were significantly increased(P<0.05 or P<0.01),the activities of T-AOC and SOD in the cells in different doses of 10-HDA groups were significantly decreased(P<0.05 or P<0.01).Compared with control group,the expression level of Bcl-2 protein in the cells in low dose of 10-HDA group was significantly decreased(P<0.01),and the expression level of GSK3β protein was significantly increased(P<0.05);compared with control group,the expression levels of Bax,Caspase-3,Caspase-9,and GSK3β proteins in the cells in middle and high doses of 10-HDA groups were significantly increased(P<0.05 or P<0.01),and the expression levels of Bcl-2,β-catenin,and CyclinD1 proteins were significantly decreased(P<0.01).Conclusion:10-HDA can significantly inhibit the proliferation and migration of the colon cancer cells and promote the apoptosis and oxidation levels of the colon cancer cells,and its mechanism may be related to the activation of the GSK3β/β-catenin signaling pathway.

BACKGROUND:The biological behavior of mesenchymal stem cells is influenced by the survival microenvironment.Pre-treatment of the microenvironment is an important means of regulating the function of mesenchymal stem cells. OBJECTIVE:To compare the differences in paracrine function of umbilical cord mesenchymal stem cells and adipose mesenchymal stem cells under oxidative stress and hypoxia,and to provide a theoretical basis for selecting appropriate pretreatment of mesenchymal stem cells to treat different diseases. METHODS:Umbilical cord mesenchymal stem cells and adipose mesenchymal stem cells were cultured by H2O2 or O2 oxygen,respectively,and cell morphology,proliferation,viability and paracrine factor expression were examined. RESULTS AND CONCLUSION:(1)The expression levels of brain-derived neurotrophic factor and transforming growth factor-β were higher in umbilical cord mesenchymal stem cells than in adipose mesenchymal stem cells under a normal culture environment,while the expressions of stromal cell-derived factor-1α and tumor necrosis factor stimulating factor-6 in the adipose mesenchymal stem cells were significantly higher than those in the umbilical cord mesenchymal stem cells.(2)There was no significant difference in the effect of low and moderate levels(≤100 μmol/L)of H2O2 on the viability of the two mesenchymal stem cells.However,increasing the H2O2 concentration from 50 μmol/L to 100 μmol/L resulted in a distinct increase in vascular endothelial growth factor expression in umbilical cord mesenchymal stem cells.The expression of basic fibroblast growth factor,vascular endothelial growth factor,stromal cell-derived factor-1α and interleukin-10 in adipose mesenchymal stem cells was greatly increased by increasing H2O2 concentration in this range.(3)1%O2 hypoxia promoted mesenchymal stem cell proliferation.After 24 hours of culture in 1%O2,gene expression levels were elevated in both mesenchymal stem cells,but the expression levels of vascular endothelial growth factor,interleukin-10 and tumor necrosis factor stimulating factor-6 were significantly higher in adipose mesenchymal stem cells than in umbilical cord mesenchymal stem cells.(4)It is concluded that hypoxia and oxidative stress preconditioning enhances the paracrine function of mesenchymal stem cells.However,mesenchymal stem cells respond differently to hypoxia and oxidative stress.Treating diseases can choose suitable mesenchymal stem cells for appropriate pretreatment to further enhance their therapeutic potential.

Objective To observe the improvement effect and mechanism of Chaishao Ruzeng capsule on mammary gland hyperplasia model rats with liver stagnation and qi stagnation.Methods The rats were injected with estradiol benzoate and unpredictable stress stimulation to replicate the model of mammary hyperplasia with liver depression and qi stagnation.Model rats were given high,medium and low doses of Chaishao Ruzeng Capsule(1.2,0.6,0.3 g·kg-1).28 consecutive days.The general state and behavioral indexes,nipple diameter and height,and pathomorphological changes of breast tissue were observed,and serum P,LH,FSH,E2,5-HT,NE,DA,MTL,VIP,SP were measured.Results Compared with the blank control group,the rats in the model group were depilated,lassitude,unresponsive,irritability,less spontaneous activity,lower sucrose preference(P<0.01),and lower body weight;The nipple diameter and height both increased(P<0.01);The degree of hyperplasia of breast lobule increased(P<0.01).The contents of E2,LH,5-TH,NE,DA,MTL and SP in serum increased,while the contents of P and VIP decreased(P<0.01).Compared with the model group,the symptoms of depilation,lassitude,unresponsiveness and irritability were significantly improved in the various dosage groups of Chaishao Ruzeng capsule,the number of autonomous activities and sucrose preference increased(P<0.05 or P<0.01).The nipple diameter and height both decreased(P<0.05 or P<0.01);The contents of P,FSH and VIP in serum were significantly increased,and the contents of E2,LH,5-TH,NE,DA,MTL and SP were significantly decreased(P<0.05 or P<0.01).Conclusions Chaishao Ruzeng capsule can alleviate sexual hormone disorder,regulate the level of neurotransmitters,improve gastrointestinal hormone abnormalities,improve liver-qi stagnation symptoms,play a role in inhibiting mammary gland hyperplasia.

Objective To explore the clinical efficacy of ginseng schisandra decoction in children with lobar pneumonia(lung spleen deficiency syndrome)induced by mycoplasma pneumoniae infection and improvement of inflammatory indicators.Method 84 children with lobar pneumonia(lung spleen deficiency syndrome)caused by mycoplasma infection admitted to the First Clinical Hospital of Jilin Academy of Traditional Chinese Medicine from January 2020 to July 2022 were selected and randomly divided into two groups with 42 cases each.The conventional group was treated according to the traditional treatment plan,while the combined group was treated with ginseng and schisandra decoction in addition to the traditional treatment plan.After 7 days,compare the differences in traditional Chinese medicine syndrome scores,clinical efficacy,inflammatory indicators,and total incidence of adverse reactions between the two groups of children.Result The results showed that the traditional Chinese medicine syndrome score and inflammatory indicators in the combined group were lower than those in the conventional group(P<0.05),and the clinical efficacy level distribution was better than that in the conventional group(P<0.05).The total effective rate was higher than that in the conventional group(P<0.05),and there was no difference in the total incidence of adverse reactions between the two groups(P>0.05).Conclusion In the treatment of children with lobar pneumonia(lung spleen deficiency syndrome)caused by Mycoplasma pneumoniae infection,the modified Ginseng Wuweizi decoction has a positive therapeutic effect and good safety,and is worth promoting.

OBJECTIVE:To interpret the record of set prescription preparations containing Panax ginseng in 2015 edition of Chinese Pharmacopoeia (part Ⅰ),and to provide reference for the future research and new drug development of P ginseng.METHODS:The set prescription preparations containing P.ginseng in 2015 edition of Chinese Pharmacopoeia (part Ⅰ) were collected to classify and analyze the selection,dosage form,preparation method,functions of curing and precautions,etc.RESULTS & CONCLUSIONS:In 2015 edition of Chinese Pharmacopoeia (part Ⅰ),there were 116 kinds of set prescription preparations containing P.ginseng,P ginseng and Radix Ginseng Rubm were the mainly selected,and combined with other Ginseng.The ingredients of set prescription preparations were mainly below fifteen ingredients;dosage forms were mainly pills (34 kinds) and capsules (28 kinds);oral administration was used as the main usage (114 kinds);main preparation method was that P ginseng was smashed into fine powder and used directly as medicine (68 kinds);the functions of curing included tonifying qi-blood,nourishing yin and tonifying kidney,clearing heat and resolving phlegm,nourishing lung and nourishing heart.In the future research,researchers will explore other effects of P.ginseng or P.ginseng combined with other drugs,and develop new drugs containing P.ginseng according to set prescription preparations containing P.ginseng in 2015 edition of Chinese Pharmacopoeia (part Ⅰ).