1.Expert Consensus on Clinical Application of Qidong Yixin Oral Liquid
Changkuan FU ; Xiaochang MA ; Mingjun ZHU ; Yue DENG ; Hongxu LIU ; Mingxue ZHANG ; Ying CHEN ; Yan ZHOU ; Ling ZHANG ; Jianhua FU ; Wei YANG ; Yu'er HU ; Ming CHEN ; Yanming XIE ; Yuanyuan LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):147-158
The prescription of Qidong Yixin oral liquid is derived from the experience of national medical master Ren Jixue in treating viral myocarditis (VMC). It has the functions of tonifying Qi, nourishing the heart,calming the mind, and relieving palpitations. It is used to treat VMC and angina pectoris of coronary heart disease caused by deficiency of both Qi and Yin. However,the understanding of its efficacy evidence, advantageous aspects, dosage and administration, and medication safety remains insufficient in clinical practice. Therefore,the development of the Expert Consensus on the Clinical Application of Qidong Yixin Oral Liquid (hereinafter referred to as consensus) was initiated. Consensus strictly followed the process and methods of the expert consensus on the clinical application of Chinese patent medicines of the China Association of Chinese Medicine,successively completing multiple tasks such as the consensus project initiation,determination of clinical problems,evidence search and evaluation,formation of recommendation opinions and consensus suggestions,solicitation of opinions,peer review, submission for review and release, and so on. Consensus formed a total of 10 recommendation opinions and 12 consensus suggestions,clarifying the clinical positioning,efficacy advantages,syndrome differentiation,dosage and administration,combination therapy,timing of medication,adverse reactions,contraindications, and precautions of Qidong Yixin oral liquid,indicating that it has good clinical advantages and safety in the treatment of VMC and angina pectoris of coronary heart disease,providing norms and references for physicians to safely and rationally apply Qidong Yixin oral liquid. Consensus was reviewed and approved for release by the Standardization Office of the China Association of Chinese Medicine on December 23, 2024. Standard number:GSCACM-376-2024.
2.Regulation of NLRP3 Inflammasome by Traditional Chinese Medicine in Treatment of Atopic Dermatitis: A Review
Minmin HU ; Aimin LIU ; Mengying MA ; Changyu WU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):315-321
Atopic dermatitis (AD) is an atopic disease with a complex etiology and pathogenesis resulting from the interaction of multiple factors. The NOD-like receptor pyrin domain-containing 3 (NLRP3) inflammasome is an important component of innate immunity and is involved in the onset and progression of AD, encompassing multiple processes such as inflammation, pyroptosis, and autophagy. Traditional Chinese medicine (TCM) has shown significant clinical efficacy in the treatment of AD and also offers advantages including flexible compatibility, multi-target effects, and low drug resistance. A large number of studies have shown that single Chinese medicinal components and compound prescriptions can treat atopic diseases by modulating the NLRP3 inflammasome. This article elaborates on the activation of the NLRP3 inflammasome and its influence on the pathogenesis and progression of AD, and summarizes recent studies on the mechanisms by which active constituents, extracts, and compound formulations of Chinese medicine treat AD through regulation of the NLRP3 inflammasome and related signaling pathways, with the aim of providing a reference for the clinical treatment of AD and the development of TCM.
3.Prediction and verification of the mechanism of Chaiqi yigan granules improving hepatocellular carcinoma
Guiping MA ; Yuanjie ZHANG ; Yichi ZHOU ; Jinzhen LYU ; Conghui WANG ; Fenping LU ; Bowen LIU ; Yun RAN ; Shiping HU
China Pharmacy 2026;37(5):620-625
OBJECTIVE To predict and validate the mechanisms of Chaiqi yigan granules (CQYG) improving hepatocellular carcinoma (HCC). METHODS The signaling pathways of CQYG intervention in HCC were predicted using network pharmacology. A mice model of transplanted hepatocellular carcinoma was established by injecting H22 hepatoma cells into the axilla. Successfully modeled mice were randomly divided into model group (normal saline), sorafenib group (positive control, 50 mg/kg), and CQYG low-, medium- and high-dose groups (24.83, 49.66, 99.32 g/kg), with 10 mice in each group. Mice in each group were administered the corresponding drug solution or normal saline intragastrically, once a day, for 14 consecutive days. After last administration, pathological morphological changes in the tumor tissues of mice were observed in each group. Immunohistochemical staining was performed to detect the expression of the nuclear proliferation antigen Ki-67 in tumor tissues of mice. Western blot assay was used to measure the expression of proteins related to epithelial-mesenchymal transition (EMT) [N-cadherin, E-cadherin, Vimentin, matrix metalloproteinase 7 (MMP7)] and the mitogen-activated protein kinase (MAPK) signaling pathway [p38 MAPK, phosphorylated p38 MAPK, c-Jun N-terminal kinase (JNK), phosphorylated JNK, extracellular regulated protein kinase 1/2 (ERK1/2), phosphorylated ERK1/2] in tumor tissue of mice. RESULTS Network pharmacology analysis revealed that metabolic pathways, pathways in cancer, and the MAPK signaling pathway were key signaling pathways through which CQYG exert their anti-hepatocellular carcinoma effects. In animal experiments, the tumor tissues of mice in the model group exhibited dense tumor cells and vigorous growth. Compared with model group, CQYG high-dose group showed a decreased density of tumor cells in the tumor tissues of mice. Moreover, the expression levels of Ki-67, N-cadherin, MMP7 and Vimentin proteins, along with the phosphorylation levels of ERK1/2 and JNK proteins, were all significantly reduced ( P <0.05). The expression level of E-cadherin protein was significantly increased ( P <0.05), the phosphorylation level of p38 MAPK protein was increased, the difference was not statistically significant ( P >0.05). CONCLUSIONS CQYG can inhibit EMT by regulating the MAPK signaling pathway, thereby suppressing tumor cell invasion and metastasis and ultimately exerting a therapeutic effect in improving HCC.
4.Comparison of sleep EEG power spectral density between depressive episode patients and schizophrenia patients with suicidal behavior
Jingwen LIU ; Yunfei ZHOU ; Jingchu HU ; Jiaoyan ZHOU ; Junwei YANG ; Jie LIANG ; Hong XU ; Yu CANG ; Shimeng MA
Sichuan Mental Health 2026;39(1):50-57
BackgroundPatients with depressive episode and schizophrenia have a high risk of suicide. The sleep electroencephalogram power spectral density characteristics of patients with depressive episode accompanied by suicidal behavior and those with schizophrenia may be different, but there is currently a lack of direct comparative studies on these two groups of patients. ObjectiveTo compare the sleep electroencephalogram power spectral density between depressive episode and schizophrenic patients with suicidal behavior, in order to provide references for exploring predictive indicators of suicidal behavior. MethodsFrom June 2018 to December 2020, 20 patients with depressive episode and 20 patients with schizophrenia who had committed suicide within the past month and were treated at the outpatient department of Shenzhen Kangning Hospital were selected. All of them met the diagnostic criteria for depressive episode or schizophrenia as defined in the International Classification of Diseases, tenth edition (ICD-10). Using a random sampling method, 20 volunteers with matching gender and age to the patient groups were selected from the Cuiping community in Shenzhen as the control group. The subjective sleep of the patients was evaluated using the Insomnia Severity Index (ISI), the Dysfunctional Belief and Attitude about Sleep (DBAS), the Disturbing Dreams and Nightmare Severity Index (DDNSI), and the Epworth Somnolence Scale (ESS). The objective sleep of the patients was assessed using polysomnography. The sleep electroencephalogram was filtered and the power spectral density of the brain wave was analyzed and processed for all the subjects. The subjective and objective sleep conditions of the two patient groups were compared, and the sleep electroencephalogram power spectral density of the patient groups and the control group were also compared. ResultsA comparison of subjective and objective sleep conditions between patients with depressive episode accompanied by suicidal behavior and patients with schizophrenia accompanied by suicidal behavior showed no statistically significant differences (P>0.05). Comparisons of sleep electroencephalogram power spectral density in the W stage (average power of α wave, total power of δ wave, average power of δ wave, average power of θ wave), N1 stage (average power of β wave, total power of α wave, total power of δ wave), N2 stage (total power of α wave, average power of α wave, total power of δ wave, average power of δ wave), N3 stage (average power of α wave, average power of δ wave), and R stage (total power of α wave, average power of α wave, total power of δ wave, average power of δ wave) between patients with depressive episode accompanied by suicidal behavior, patients with schizophrenia accompanied by suicidal behavior, and the control group showed statistically significant differences (P<0.05 or 0.01). The total power of δ wave in the W stage and the average power of β wave and δ wave in the N1 stage were higher in two patient groups were higher than those of the control group. The total power of α wave and the average power of α wave in the N2 stage were lower than those of the control group, while the average power of δ wave was higher than that of the control group. The average power of α wave in the N3 stage of both patient groups were lower than that of the control group, while the average power of δ wave was higher than that of the control group. The total power and average power of α wave in the R stage were lower than those of the control group, while the total power and average power of δ wave were higher than those of the control group. All the differences were statistically significant. Patients with depressive episode accompanied by suicidal behavior had higher average powers of α wave, δ wave, and θ wave in the W stage compared with the control group, while the total power of α wave in the N1 stage was lower in the former group. All these differences were statistically significant (P<0.05). ConclusionThe depressive episode patients accompanied by suicidal behavior have highly overlapping sleep electroencephalogram abnormal patterns with those of schizophrenia patients, mainly manifested as a general decrease in α wave power (N2, N3, R stage) and a general increase in δ wave power (W, N1, N2, N3, R stage) as well as β wave power in N1 stage. At the same time, patients with depressive episode accompanied by suicidal behavior also show specific changes, including an increase in the average power of α and θ waves during the wakefulness period (W stage), and a decrease in the total power of α wave in N1 stage. [Funded by Guangdong Province High-level Clinical Key Specialty (with supporting funds from Shenzhen City) (number, SZGSP013); Shenzhen Key Medical Discipline (number, SZXK041); Shenzhen Clinical Medicine Research Center Project (number, 20210617155253001)]
5.Rapid Discrimination of Processing Degree of Wine-processed Chuanxiong Rhizoma Based on Intelligent Sensory Technology and Multivariate Statistical Analysis
Xiaolong ZHANG ; Xiaoni MA ; Xinzhu WANG ; Po HU ; Yang PAN ; Tulin LU ; Guangming YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):174-182
ObjectiveTo explore the changes in color, odor and chemical components during wine-processing of Chuanxiong Rhizoma(CR), identify differential markers, and provide a basis for standardizing the process and establishing quality standards. MethodsFifteen batches of CR samples from 4 producing areas were collected. Colorimeter and electronic nose were used to detect the color changes and odor components of CR before and after wine-processing. Multivariate statistical methods including partial least squares-discriminant analysis(PLS-DA), principal component analysis(PCA), discriminant factor analysis(DFA) and Fisher discriminant analysis were applied to identify wine-processed CR at different processing stages and establish discriminant models, and differential components were screened out based on variable importance in the projection(VIP) value1. Then, high performance liquid chromatography(HPLC) was employed to detect the content changes of four components(ferulic acid, senkyunolide I, senkyunolide A and ligustilide) during the processing stages. ResultsThe differences of wine-processed CR at various stages were primarily reflected in color parameters L*(brightness value), a*(red-green value) and b*(yellow-blue value). Based on chromaticity differences, the color reference ranges were established for moderately processed CR, including L* of 46.75-48.24, a* of 5.37-6.07 and b* of 20.32-21.70. In odor analysis, DFA revealed significant differences among processing stages, and 11 odor markers were identified, with four differential markers(4-hydroxy-3-butylphthalide, isopropyl butyrate, L-limonene and 1-methoxyhexane) based on VIP values. HPLC results showed that there was no significant difference of the four components except for ligustilide in wine-processed CR at different stages. ConclusionThis study achieved rapid identification of wine-processed CR with different processing degrees by electronic sensory technology and differential component content detection, with discrimination accuracy rates of 92.4% and 93.272% for color and odor, respectively. This paper also established the reference ranges of main colorimetric parameters for wine-processed CR at different stages, and four differential components were screened out, providing a basis for standardizing the processing of wine-processed CR and establishing quality standards for this decoction pieces.
6.Rapid Discrimination of Processing Degree of Wine-processed Chuanxiong Rhizoma Based on Intelligent Sensory Technology and Multivariate Statistical Analysis
Xiaolong ZHANG ; Xiaoni MA ; Xinzhu WANG ; Po HU ; Yang PAN ; Tulin LU ; Guangming YANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):174-182
ObjectiveTo explore the changes in color, odor and chemical components during wine-processing of Chuanxiong Rhizoma(CR), identify differential markers, and provide a basis for standardizing the process and establishing quality standards. MethodsFifteen batches of CR samples from 4 producing areas were collected. Colorimeter and electronic nose were used to detect the color changes and odor components of CR before and after wine-processing. Multivariate statistical methods including partial least squares-discriminant analysis(PLS-DA), principal component analysis(PCA), discriminant factor analysis(DFA) and Fisher discriminant analysis were applied to identify wine-processed CR at different processing stages and establish discriminant models, and differential components were screened out based on variable importance in the projection(VIP) value1. Then, high performance liquid chromatography(HPLC) was employed to detect the content changes of four components(ferulic acid, senkyunolide I, senkyunolide A and ligustilide) during the processing stages. ResultsThe differences of wine-processed CR at various stages were primarily reflected in color parameters L*(brightness value), a*(red-green value) and b*(yellow-blue value). Based on chromaticity differences, the color reference ranges were established for moderately processed CR, including L* of 46.75-48.24, a* of 5.37-6.07 and b* of 20.32-21.70. In odor analysis, DFA revealed significant differences among processing stages, and 11 odor markers were identified, with four differential markers(4-hydroxy-3-butylphthalide, isopropyl butyrate, L-limonene and 1-methoxyhexane) based on VIP values. HPLC results showed that there was no significant difference of the four components except for ligustilide in wine-processed CR at different stages. ConclusionThis study achieved rapid identification of wine-processed CR with different processing degrees by electronic sensory technology and differential component content detection, with discrimination accuracy rates of 92.4% and 93.272% for color and odor, respectively. This paper also established the reference ranges of main colorimetric parameters for wine-processed CR at different stages, and four differential components were screened out, providing a basis for standardizing the processing of wine-processed CR and establishing quality standards for this decoction pieces.
7.Clinical application value of intracavitary PRP infusion combined with IVF-FET in patients with chronic endometritis
Xiaotong ZHANG ; Xiaoyuan HAO ; Rui FANG ; Shuyao HU ; Linkun MA ; Yaqi ZHAO ; Wei HAN
Chinese Journal of Blood Transfusion 2025;38(3):382-387
[Objective] To evaluate the clinical application value of intrauterine perfusion with platelet-rich plasma (PRP) combined with in vitro fertilization-frozen-thawed embryo transfer (IVF-FET) in patients with chronic endometritis (CE). [Methods] A randomized controlled trial (RCT) was conducted, enrolling 60 CE patients undergoing artificial cycle frozen embryo transfer at our hospital from January 2022 to January 2024. Participants were randomly divided into three groups: Group A (routine frozen embryo transfer, n=20), Group B (routine frozen embryo transfer + one PRP intrauterine perfusion, n=20), and Group C (routine frozen embryo transfer + two PRP intrauterine perfusions, n=20). Endometrial thickness during the transformation and transplantation phases, uterine artery pulsatility index (PI), resistance index (RI), systolic peak velocity/end-diastolic velocity (S/D) ratio during transplantation, serum levels of IL-2, IL-4, IL-6, IL-10, and TNF-α during transplantation, as well as biochemical pregnancy rate, clinical pregnancy rate, live birth rate, and early miscarriage rate were compared across groups. [Results] No significant differences in endometrial thickness were observed among the three groups during the transformation phase (P>0.05). During the transplantation phase, endometrial thickness in Groups C and B was significantly higher than in Group A[9.54 (8.96-10.22) and 8.90 (8.34-9.72) vs 8.37 (7.89-8.75) mm, P<0.05], with Group C showing greater thickness than Group B (Z=3.733, P<0.05). Endometrial thickness in Groups C and B during transplantation was significantly increased compared to their respective transformation phases (Z=2.191, 2.462; P<0.05). Groups C and B exhibited lower PI, RI, and S/D values than Group A[PI:1.87 (1.77-1.97), 1.94 (1.88-2.15) vs 2.43 (2.35-2.49); RI:0.75 (0.73-0.77), 0.78 (0.75-0.81) vs 0.84 (0.83-0.86); S/D:2.61 (2.33-3.42), 3.01 (2.20-3.93) vs 3.72 (3.06-4.49); P<0.05]. Group C demonstrated lower PI and RI than Group B (P<0.05). IL-2 levels in Groups C and B were higher than in Group A[3.88 (2.71-5.01), 3.59 (2.73-4.38) vs 3.16 (2.11-3.25) ng/L, P<0.05], while IL-4, IL-6, IL-10, and TNF-α levels were significantly lower (IL-4: Z=1.428, 2.421; IL-6: Z=1.754, 2.435; IL-10: Z=1.754, 2.854; TNF-α: Z=1.961, 1.765; P<0.05). Group C had lower IL-6 levels than Group B (Z=3.976, P<0.05). Biochemical pregnancy rate, clinical pregnancy rate, and live birth rate in Group C were significantly higher than in Group A (75% vs 40%, 70% vs 35%, 60% vs 20%, P<0.05). No significant differences in early miscarriage rates were observed among the groups (χ2=3.750, P>0.05). [Conclusion] Intrauterine autologous PRP perfusion in CE patients enhances pregnancy and live birth rates, improves pregnancy outcomes post-FET, and demonstrates superior efficacy in endometrial repair and receptivity with two PRP perfusions compared to a single perfusion. This provides a safe and effective therapeutic option for optimizing outcomes in CE patients with prior implantation failure.
8.Cognitive Disorders Awareness and Associated Risk Factors in Xizang Autonomous Region
Yu HAO ; Junshan WANG ; Ma ZHUO ; Quzhen SUOLANG ; Shiyong JI ; Yaxiong HU ; Zhijie DING ; Zhuoga CIDAN ; Jing YUAN ; Yuhua ZHAO
Medical Journal of Peking Union Medical College Hospital 2025;16(2):472-478
To investigate the awareness of cognitive impairment disorders among residents of the Xizang Autonomous Region and its influencing factors, thereby providing a basis for targeted prevention and treatment efforts. From April to December 2024, a questionnaire survey was conducted among permanent residents aged ≥18 years (residing in the Xizang Autonomous Region for 180 days or more). The survey was primarily conducted online, supplemented by QR code distribution during community medical outreach by healthcare workers. Demographic information and data on awareness of cognitive disorders were collected, and an ordered Logistic regression model was used to analyze influencing factors in the overall population and stratified by occupation. A total of 327 questionnaires were collected, with 14 excluded (13 for not meeting residency requirements and 1 for self-reported diagnosis of cognitive impairment), leaving 313 valid questionnaires. The average age of respondents was 42.0±11.9 years; 108 (34.5%) were male, and 205 (65.5%) were female. Most respondents were from Lhasa (78.6%, 246/313); 179 (57.2%) were healthcare workers, and 134 (42.8%) were non-healthcare workers. Regarding awareness of cognitive impairment disorders, 7.3% (23/313) were "unaware", 75.7% (237/313) were "partially aware", and 16.9% (53/313) were "well aware".Ordered Logistic regression analysis revealed that education level of high school or below ( Awareness of cognitive impairment disorders among residents of the Xizang Autonomous Region needs improvement. Educational level, occupation, and prior contact with cognitive impairment patients significantly influence disease awareness. Enhancing overall education levels and using vivid clinical case presentations in health education and public outreach are key strategies to improve public awareness of cognitive impairment disorders.
9.Mechanism of IGF2BP2 regulation of PPAR-γ/GLUT4 pathway in insulin resistance induced by sodium arsenite exposure in HepG2 cells
Shiqing XU ; Zhida HU ; Qiyao ZHANG ; Siqi ZHAO ; Yujie WANG ; Xiaohui WANG ; Teng MA ; Li WANG
Journal of Environmental and Occupational Medicine 2025;42(4):400-407
Background Arsenic is an environmentally harmful substance that causes hepatic insulin resistance and liver damage, increasing the risk of type 2 diabetes mellitus. Objective To explore whether the insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) is involved in insulin resistance in HepG2 cells after arsenic exposure through the peroxisome-proliferator-activated receptor γ (PPAR-γ) / glucose transporter 4 (GLUT4) pathway. Methods Cell viability was determined using cell counting kit 8 (CCK8) and an appropriate NaAsO2 infection dose was determined. A cellular arsenic exposure model of HepG2 cells was established by four concentrations of NaAsO2 solution for 24 h (the experiment was divided into four groups: 0, 2, 4, and 8 μmol·L−1); HepG2 cells were firstly treated with pcDNA3.1-IGF2BP2 and pcDNA3.1-NC respectively for 6 h, then with 8 μmol·L−1 NaAsO2 for 24 h to establish a IGF2BP2 overexpression cell model (the experiment was divided into 4 groups: control, NaAsO2, NaAsO2+pcDNA3.1-IGF2BP2, and NaAsO2+pcDNA3.1-NC); finally the cells were subject to 100 nmol·L−1 insulin stimulation for 30 min. Glycogen and glucose in HepG2 cells were determined by glycogen and glucose assay kits; mRNA expression levels of IGF2BP2 were measured by quantitative real-time PCR; protein expression levels of IGF2BP2, PPAR-γ, and GLUT4 in HepG2 were detected by Western blot (WB); and the binding of IGF2BP2 to PPAR-γ and PPAR-γ to GLUT4 was verified by co-immunoprecipitation (CO-IP) experiment. Results The results of CCK8 experiment showed a dose-effect relationship between NaAsO2 concentration and cell viability. When the concentration of NaAsO2 was ≥4 μmol·L−1 , the cell viabilities were lower than that of the control group (P <0.05). With the increasing dose of NaAsO2 infection, reduced glucose consumption and glycogen levels in HepG2 cells were found in the 2, 4, and 8 μmol·L−1 NaAsO2 treatment groups compared to the control group (P <0.05). The difference between the mRNA expression level of IGF2BP2 in the HepG2 cells treated with 4 or 8 μmol L−1 NaAsO2 and the control group was significant (P <0.05). In the IGF2BP2 overexpression cell model, compared with the control group, glucose consumption and glycogen levels were lowered in the NaAsO2 group (P <0.05), the mRNA expression level of IGF2BP2 and the protein expression levels of IGF2BP2, PPAR-γ, and GLUT4 in the cell membrane were all decreased (P <0.05). Compared with the NaAsO2 group, the glucose consumption and glycogen levels were increased in the NaAsO2+pcDNA3.1-IGF2BP2 group (P <0.05), and the mRNA expression level of IGF2BP2 and the protein expression levels of IGF2BP2, PPAR-γ, and GLUT4 in the cell membrane were all increased (P <0.05). The results of CO-IP experiments showed that IGF2BP2 interacted with PPAR-γ as well as PPAR-γ with GLUT4 protein. Conclusion IGF2BP2 is involved in arsenic exposure-induced insulin resistance in HepG2 cells by acting on the PPAR-γ/GLUT4 pathway.
10.Anthocyanins from Lycium ruthenicum Murr combined with human adipose-derived pericytes/perivascular cells support proliferation of umbilical cord blood hematopoietic stem/progenitor cells
Yamei SHEN ; Yunxia NIU ; Tingting YANG ; Jie MA ; Daihong HU ; Bo ZHENG
Chinese Journal of Tissue Engineering Research 2025;29(1):58-64
BACKGROUND:Anthocyanin is one of the most important active components in Lycium ruthenicum Murr,which has antioxidant and immunomodulatory effects.CD146+human adipose-derived pericytes/perivascular cells(CD146+hAD-PCs)are the progenitors of bone marrow mesenchymal stem cells,which can promote the proliferation and differentiation of hematopoietic stem/progenitor cells in vitro.The support effect of anthocyanin in combination with CD146+hAD-PCs on umbilical cord blood hematopoietic stem/progenitor cells remains to be studied. OBJECTIVE:To investigate the supporting effect of anthocyanins in Lycium ruthenicum Murr(ALRM)combined with CD146+hAD-PCs on umbilical cord blood CD34+hematopoietic stem/progenitor cells(UCB CD34+HSPCs)in vitro. METHODS:The CCK-8 assay was used to detect the effect of different concentrations(0,200,400,600,800,1 000 mg/L)of ALRM on the proliferation of CD146+hAD-PCs.Flow cytometry was used to detect the effect of ALRM on the cell cycle of CD146+hAD-PCs.The co-culture experiments were divided into blank group,ALRM group,CD146+hAD-PCs group,and ALRM+CD146+hAD-PCs group to analyze the in vitro supporting effect of ALRM combined with CD146+hAD-PCs on UCB CD34+HSPCs.The number of expanded cells and the number of colony-forming units were compared at 1,2,and 4 weeks of co-culture.The immunophenotype of cells was detected by flow cytometry.The level of cytokines was detected by enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION:(1)The cell viability of CD146+hAD-PCs was highest at an ALRM concentration of 200 mg/L,the proportion of G0/G1 phase cells decreased and the proportion of S and G2/M phase cells increased in CD146+hAD-PCs(P<0.01).(2)The change in number of UCB CD34+HSPCs cells in the ALRM+CD146+hAD-PCs group was higher than that in the ALRM group at 1,2,and 4 weeks of co-culture(all P<0.05),and higher than that in CD146+hAD-PCs group at 2 and 4 weeks of co-culture(all P<0.05).The number of cells in the ALRM group and blank group decreased gradually with the extension of co-culture time.(3)Colony forming capacity and immunophenotype analysis:The number of colony-forming units in the ALRM+CD146+hAD-PCs group was higher than that in the CD146+hAD-PCs group and ALRM group at 1 and 2 weeks of co-culture(P<0.05).The proportion of CD45+and CD34+CD33-cells in the ALRM+CD146+hAD-PCs group was higher than that in the CD146+hAD-PCs group at 1 and 2 weeks of co-culture(all P<0.01).(4)Changes in cytokines:Interleukin-2 level in the ALRM+CD146+hAD-PCs group was higher than that in the ALRM and CD146+hAD-PCs groups(P<0.05).The interleukin-3 content of the ALRM+CD146+hAD-PCs group was higher than that of the CD146+hAD-PCs group at 2 and 4 weeks(P<0.05).The expression level of granulocyte colony-stimulating factor in the ALRM+CD146+hAD-PCs group was higher than that in the CD146+hAD-PCs group at 1 week,and higher than that in the ALRM group and CD146+hAD-PCs group at 2 weeks(P<0.01).Interferon-γ content in the ALRM group and ALRM+CD146+hAD-PCs group was lower than that in the CD146+hAD-PCs group at 1,2,and 4 weeks of co-culture(P<0.01).(5)Due to the absence of stromal cells in the blank group,UCB CD34+HSPCs could not be counted after 1 week of co-culture and were not subjected to immunophenotyping,colony analysis,or cytokine assays.(6)In summary,ALRM can promote the expansion of UCB CD34+HSPCs in vitro by promoting CD146+hAD-PCs proliferation and cell cycle transformation,which is of great value in hematopoietic stem cell transplantation.

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