ObjectiveTo reveal the active substances and mechanisms of modified Qing'e formula （MQEF） in delaying skin photoaging by ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry （UPLC-Q-TOF-MS），network pharmacology， and cell experiments. MethodsUPLC-Q-TOF-MS and a literature review were employed to analyze the transdermally absorbed components in mice after the topical application of MQEF. The potential targets of MQEF in treating skin photoaging were retrieved from databases.The compound-potential target network and protein-protein interaction network were constructed to screen the key components and core targets. A photoaging cell model was established by irradiating HaCaT cells with medium-wave ultraviolet B （UVB）. The safe doses of bakuchiol （BAK） and salvianolic acid B （SAB） for treating HaCaT cells and the effects of BAK and SAB on the viability of cells exposed to UVB irradiation were determined by the cell counting kit-8 （CCK-8） method.The reactive oxygen species （ROS） fluorescent probe was used to measure the ROS production in the cells treated with BAK and SAB.The expression levels of genes related to oxidative stress，inflammation，collagen metabolism，and circadian rhythm clock were measured by Real-time PCR. ResultsA total of 24 transdermally absorbed components of MQEF were identified，which acted on 367 potential targets，and 417 targets related to skin photoaging were screened out，among which 47 common targets were predicted as the targets of MQEF in treating skin photoaging. MQEF exerted the anti-photoaging effect via key components such as BAK and SAB，which acted on core proteins such as serine/threonine kinase 1 （Akt1） and mitogen-activated protein kinase 3 （MAPK3） and intervened in core pathways such as the tumor necrosis factor （TNF） and phosphatidylinositol-3-kinase （PI3K）/protein kinase B （Akt） signaling pathways.Compared with the model group，the administration of BAK and SAB increased the survival rate of HaCaT cells （P<0.01），down-regulated the mRNA levels of cyclooxygenase-2 （COX-2），interleukin-6 （IL-6），tumor necrosis factor-α （TNF-α），matrix metalloproteinase-1 （MMP-1），and matrix metalloproteinase-9 （MMP-9） （P<0.01），and up-regulated the mRNA levels of heme oxygenase-1 （HO-1） and NAD（P）H quinone dehydrogenase 1 （NQO-1） （P<0.05，P<0.01） in photoaged HaCaT cells.In addition，it eliminated excess ROS production induced by UVB and up-regulated the mRNA levels of brain and muscle ARNT-like 1 （BMAL1） and circadian locomotor output cycles kaput （CLOCK） associated with circadian clock （P<0.05，P<0.01）. ConclusionMQEF delays skin photoaging through the coordinated effects of various components，multiple targets，and diverse pathways.The key components BAK and SAB in MQEF exhibit anti-photoaging properties，which involve inhibiting oxidative stress，preventing collagen degradation，mitigating inflammation，and maintaining normal skin circadian rhythms by regulating clock gene expression.

Coronary heart disease is a chronic and lifelong disease， which places a dual burden on the physiological and psychological well-being of patients， and can easily lead to psychological distress and affect their prognosis and quality of life. This article provides a systematic review， in which the current status， evaluation tools， influencing factors and intervention methods of psychological distress in patients with coronary heart disease are explored， aiming to provide key information beneficial for identifying and preventing psychological distress， and to improve the overall management and treatment effectiveness of coronary heart disease patients. In this paper， 18 articles were included， and the demographic， physiological， psychological and social factors affecting the psychological distress of patients with coronary heart disease were systematically analyzed， thus to provide a deeper understanding of psychological distress and offering references for formulating targeted intervention strategies.

OBJECTIVES: To reprogram human placental fibroblasts (HPFs) into chemically induced epithelioid-like cells (ciEP-Ls) using a combination of small-molecule compounds. METHODS: HPFs cultured under normoxic conditions were identified using immunofluorescence assay, PCR and chromosomal karyotyping. Under hypoxic conditions (37 ℃, 5% O₂), HPFs were cultured in a medium containing small-molecule compounds C6TSEDRVAJZ (CHIR99021, 616452, TTNPB, SAG, EPZ5676, DZNep, Ruxolitinib, VTP50469, Afuresertib, JNK-IN-8, and EZM0414), and the cell morphology was observed daily. The expression levels of epithelial cell markers in the induced cells were detected by immunofluorescence, Western blotting and PCR. Chromosomal karyotyping of the induced cells was performed and the induction efficiency was calculated. RESULTS: Before induction, HPFs showed positive expressions of fibroblast surface markers CD34 and vimentin and were negative for epithelial surface markers. PCR results showed high expressions of fibroblast-specific genes S100A4 and COL1A1 in HPFs with a normal human diploid karyotype. After one day of induction, the HPFs underwent morphological changes from a multinodular spindle shape to a round or polygonal shape, which was morphologically characteristic of ciEP-Ls. On day 4 of induction, the cells exhibited high expressions of the epithelial cell markers E-cadherin and Lin28A. RT-qPCR results also showed that the cells expressed the epithelial markers Smad3, GLi3, PAX8, WT1, KRT19, and KRT18 with significantly down-regulated expressions of all the fibroblast surface markers and a normal human diploid karyotype. The reprogramming efficiency of HPFs into ciEP-Ls ranged from (64.53±2.8)% to (68.10±3.6)%. CONCLUSIONS The small-molecule compound combination C6TSEDRVAJZ is capable of inducing HPFs into ciEP-Ls under hypoxic conditions with a high induction efficiency.
Humans ; Fibroblasts/drug effects* ; Pregnancy ; Female ; Cell Differentiation/drug effects* ; Pyrimidines/pharmacology* ; Placenta/cytology* ; Cells, Cultured ; Pyridines/pharmacology* ; Pyrazoles/pharmacology* ; Epithelial Cells/cytology*

Objective Based on the Ancient and Modern Medical Record Cloud Platform,the medication rules of Chinese herbal medicine for the treatment of polycystic ovary syndrome(PCOS)of liver stagnation type were explored.Methods From database inception till December 2023,the medical records of TCM in treating PCOS of liver stagnation type were retrieved from the databases of China National Knowledge Infrastructure(CNKI),China Science and Technology Journal Database(VIP),Wanfang Data Knowledge Service Platform(Wangfang).After that,the data of prescription and TCM syndrome type were extracted,and then a database was constructed.The Ancient and Modern Medical Record Cloud Platform(V2.3.7)was used to conduct statistical analysis of relevant types of liver stagnation,occurrence frequency of Chinese herbal medicines,and the properties and efficacy of Chinese herbal medicines,as well as correlation analysis and clustering analysis for Chinese herbal medicines.The core prescriptions were mined by complex network analysis.Results A total of 108 prescriptions were included in this study,involving 174 Chinese herbal medicines.The syndrome of kidney deficiency and liver stagnation was most frequently seen,and Angelicae Sinensis Radix and Bupleuri Radix were the Chinese herbal medicines used most frequently.The Chinese herbal medicines usually have the meridian tropism of liver,spleen and kidney meridians,and most of them were mild and sweet in the proterties.The Chinese herbal medicines usually have the actions of soothing liver and alleviating depression.A total of 17 herbal groups such as"Angelicae Sinensis Radix-Paeoniae Radix Alba"and"Bupleuri Radix-Paeoniae Radix Alba"were obtained after association analysis,and the cluster analysis yielded four clustering combinations of Chinese herbal medicine.The core prescription was composed of Bupleuri Radix,Paeoniae Radix Alba,Angelicae Sinensis Radix,Poria,Glycyrrhizae Radix et Rhizoma,Cuscutae Semen,Rehmanniae Radix Praeparata,Cyperi Rhizoma,and Atractylodis Macrocephalae Rhizoma.Conclusion Chinese medicine treatment for PCOS of liver stagnation type focuses on soothing liver and invigorating spleen,nourishing liver and kidney,thus to harmonize qi and blood,and thoroughfare vessel and conception vessel.

Objective To investigate the effects of modified Simiao San combined with Metronidazole Tablets on immune factors,inflammatory factors,and oxidative stress in patients with bacterial vaginosis(BV).Methods Eighty BV patients treated at Dongguan Hospital of Guangzhou University of Chinese Medicine(Dongguan Traditional Chinese Medicine Hospital)from March 2023 to April 2024 were equally randomly divided into a control group(40 cases,treated with Metronidazole Tablets)and a study group(40 cases,treated with modified Simiao San plus Metronidazole Tablets).The treatment for the two groups lasted for 7 days.Serum immunologic factors[immunoglobulin(Ig)A,IgG,and IgM],inflammatory factors[C-reactive protein(CRP),tumor necrosis factor α(TNF-α),and interleukin-6(IL-6)],and oxidative stress markers[superoxide dismutase(SOD),malondialdehyde(MDA),and glutathione peroxidase(GSH-Px)]in the two groups were measured before and after treatment.After treatment,symptom resolution time,clinical efficacy,and the incidence of total adverse reactions were compared between the two groups.Results(1)After 7 days,the total effective rate was 95.00%(38/40)in the study group versus 77.50%(31/40)in the control group(tested by chi-square test,P＜0.05).(2)The study group showed significantly shorter time to the resolution of profuse or scanty leukorrhea,odor of leukorrhea,vulvar itching,vaginal pain,and vaginal mucosal hyperemia(all P＜0.01)than the control group.(3)After treatment,both groups exhibited increased IgA,IgG,and IgM(P＜0.05)compared to those before treatment,with greater improvements in the study group(P＜0.01).(4)After treatment,serum levels of inflammatory factors of CRP,TNF-α,and IL-6 in both groups decreased(P＜0.05),with superior reductions in the study group(P＜0.01).(5)After treatment,serum levels of oxidative stress markers of SOD and GSH-Px levels rose(P＜0.05),while MDA declined(P＜0.05)in both groups,and the study group demonstrated more pronounced improvements(P＜0.01).(6)Adverse reaction rates were 5.00%(2/40)and 15.00%(6/40)in the study group and control group,respectively,the differentce being no statistical significance between the two groups(P＞0.05).Conclusion Modified Simiao San combined with Metronidazole Tablets significantly alleviates BV symptoms,improves immune function,and inhibits inflammatory response and oxidative stress,demonstrating satisfactory efficacy and high safety.

Objective To investigate the relationship between spicy diet and uremic pruritus(UP)in the patients with maintenance haemodialysis(MHD).Methods A total of 403 patients receiving the treat-ment in the blood purification center of this hospital from December 2023 to February 2024 were selected as the study subjects and grouped by the sum of the scores of frequency and degree of pepper intake.The visual analogue rating scale(VAS)was used to conduct the preliminary pruritus score in all patients,and the pa-tients with the score＞0 point conducted the multidimensional evaluation by the 14-item uremic skin pruritus scale.The blood routine and itch-related blood biochemical indexes levels of all patients were measured.Results There were 65 cases in the bland diet group,119 cases in the mild spicy diet group and 219 cases in the spicy diet group,and there was no significant difference in the number of genders between the groups(P＞0.05).There were statistically significant differences in age,dialysis age and lymphocyte count among the groups(P＜0.05).There was no statistically significant difference in the degree of pruritus among the groups(Z=9.301,P=0.157),but it was seen that the proportion of moderate and severe pruritus in the mild spicy diet group and the spicy diet group was decreased,and the proportions of no pruritus and mild pruritus showed the increasing trend.The itching score of the bland diet group was higher than that of the mildly spicy diet group and spicy diet group,and the difference was statistically significant(P＜0.05),but there was no statistically significant difference between the mild spicy diet group and spicy diet group(P＞0.05).There was a statistically significant difference in the itch score of the patients aged 40-60 years in each group(P＜0.05),and there was no statistically significant difference in the itch score between the patients aged＞60 years old and＜40 years old in each group(P＞0.05).There was no statistically significant difference in the itch-related blood biochemical indexes among the groups(P＞0.05).Conclusion The spicy diet may reduce the degree of pruritus in patients with MHD,moreover which is not affected by the age and other factors,and may be associated with lymphocyte level decrease in the patients.

Objective To improve the efficiency of induced differentiation of primitive neural epithelial cells derived from human induced pluripotent stem cells(hiPSCs-NECs)into functional midbrain dopaminergic progenitor cells(DAPs).Methods HiPSCs were cultured in mTeSRTM medium containing DMH1(10 μmol/L),SB431542(10 μmol/L),SHH(200 ng/mL),FGF8(100 ng/mL),purmorphamine(2 μmol/L),CHIR99021(3 μmol/L),and N2(1%)for 12 days to induce their differentiation into primitive neuroepithelial cells(NECs).The hiPSCs-NECs were digested with collagenase IV and then cultured in neurobasal medium supplemented with 1%N2,2%B27-A,BDNF(10 ng/mL),GDNF(10 ng/mL),AA,TGF-β,cAMP,and 1%GlutaMax in the presence of different concentrations of Rho kinase inhibitor Y27632,and the culture medium was changed the next day to remove Y27632.Continuous induction was performed until day 28 to obtain DAPs.Results Human iPSCs expressed the pluripotency markers OCT4,SOX2,Nanog,and SSEA1 and were positive for alkaline phosphatase staining.The hiPSCs-NECs were obtained on day 13 in the form of neural rosettes expressing neuroepithelial markers SOX2,nestin,and PAX6.In digested hiPSCs-NECs,the addition of 5 μmol/L Y27632 significantly promoted survival of the adherent cells,increased cell viability and the proportion of S-phase cells(P<0.01),and reduced the rate of apoptotic cells(P<0.05).On day 28 of induction,the obtained cells highly expressed the specific markers of DAPS(TH,FOXA2,NURR1,and Tuj1).Conclusion Treatment with Y27632(5 μmol/L)for 24 h significantly promotes the survival of human iPSCs-NECs during their differentiation into DPAs without affecting the cell differentiation,which indirectly enhances the efficiency of cell differentiation.

Objective:To discuss the effect of cell division cycle protein 20(CDC20)on the proliferation and cell cycle of endometrial carcinoma(EC)cells,and to clarify its mechanism.Methods:Real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods were used to detect the expression levels of CDC20 mRNA and protein in human endometrial stromal T-HESC cell and EC cells(KLE,RL95-2,ZJB-ENC1,and ECC-1 cells).The RL95-2 cells were selected for the subsequent experiments.CDC20 shRNA interference lentivirus was transfected into the RL95-2 cells and the cells were divided into control group,sh-NC group(infected with negative control lentivirus),sh-CDC20 group(infected with CDC20 shRNA interference lentivirus),sh-NC+SM04690 group(infected with negative control lentivirus followed by treatment with 64 nmol·L-1 Wnt/β-catenin signaling pathway inhibitor SM04690 for 48 h),and sh-CDC20+SM04690 group(infected with CDC20 shRNA interference lentivirus followed by treatment with 64 nmol·L-1 SM04690 for 48 h).RT-qPCR and Western blotting methods were used to detect the expression levels of CDC20 mRNA and proteins in the cells in various groups;CCK-8 method was used to detect the proliferation activities of the RL95-2 cells in various groups;BrdU assay was used to detect the percentages of BrdU positive cells in various groups;flow cytometry was used to detect the percentages of the cells at G2/M stage in various groups;Western blotting method was used to detect the expression levels of β-catenin,oncogene c-Myc,and cyclin D1 proteins in the cells in various groups.Results:Compared with T-HESC cells,the expression levels of CDC20 mRNA and protein in the KLE,RL95-2,ZJB-ENC1,and ECC-1 cells were significantly increased(P＜0.05),and the highest expression levels of CDC20 mRNA and protein were observed in RL95-2 cells.Compared with sh-NC group,the proliferation activities and percentages of the BrdU positive cells in sh-CDC20 group and sh-NC+SM04690 group were significantly decreased(P＜0.05),the percentages of the cells at G2/M phase were significantly increased(P＜0.05),and the expression levels of β-catenin,c-Myc,and cyclin D1 proteins were significantly decreased(P＜0.05).Compared with sh-CDC20 group,the proliferation activity and percentage of BrdU positive cells in sh-CDC20+SM04690 group were significantly decreased(P＜0.05),the percentage of the cells at G2/M phase was significantly increased(P＜0.05),and the expression levels of β-catenin,c-Myc,and cyclin D1 proteins in the cells were significantly decreased(P＜0.05).Conclusion:CDC20 is highly expressed in the EC cells.Silencing CDC20 may inhibit the cell proliferation by inducing G2/M phase arrest in the RL95-2 cells through the regulation of Wnt/β-catenin signal transduction.

Objective To improve the efficiency of induced differentiation of primitive neural epithelial cells derived from human induced pluripotent stem cells(hiPSCs-NECs)into functional midbrain dopaminergic progenitor cells(DAPs).Methods HiPSCs were cultured in mTeSRTM medium containing DMH1(10 μmol/L),SB431542(10 μmol/L),SHH(200 ng/mL),FGF8(100 ng/mL),purmorphamine(2 μmol/L),CHIR99021(3 μmol/L),and N2(1%)for 12 days to induce their differentiation into primitive neuroepithelial cells(NECs).The hiPSCs-NECs were digested with collagenase IV and then cultured in neurobasal medium supplemented with 1%N2,2%B27-A,BDNF(10 ng/mL),GDNF(10 ng/mL),AA,TGF-β,cAMP,and 1%GlutaMax in the presence of different concentrations of Rho kinase inhibitor Y27632,and the culture medium was changed the next day to remove Y27632.Continuous induction was performed until day 28 to obtain DAPs.Results Human iPSCs expressed the pluripotency markers OCT4,SOX2,Nanog,and SSEA1 and were positive for alkaline phosphatase staining.The hiPSCs-NECs were obtained on day 13 in the form of neural rosettes expressing neuroepithelial markers SOX2,nestin,and PAX6.In digested hiPSCs-NECs,the addition of 5 μmol/L Y27632 significantly promoted survival of the adherent cells,increased cell viability and the proportion of S-phase cells(P<0.01),and reduced the rate of apoptotic cells(P<0.05).On day 28 of induction,the obtained cells highly expressed the specific markers of DAPS(TH,FOXA2,NURR1,and Tuj1).Conclusion Treatment with Y27632(5 μmol/L)for 24 h significantly promotes the survival of human iPSCs-NECs during their differentiation into DPAs without affecting the cell differentiation,which indirectly enhances the efficiency of cell differentiation.

Objective:To explore the risk factors for lymphoproliferative disorders (PTLD) in children with thalassemia major (TM) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:This was a retrospective case-control study.A total of 482 children with TM who underwent allo-HSCT at Shenzhen Children′s Hospital between January 2020 and December 2022 were selected and classified into the PTLD and non-PTLD groups according to the occurrence of PTLD.The risk factors for PTLD after allo-HSCT in children with TM were analyzed, and the diagnostic efficiency of relevant risk factors for PTLD was analyzed by receiver operating characteristic (ROC) curve.Results:A total of 25 out of 482 patients (5.2%, 25/482) developed PTLD about 114 (54-271) days after allo-HSCT.Among them, 12 cases (12/25, 48.0%) occurred within 100 days, and 22 cases (22/25, 88.0%) occurred within 1 year after allo-HSCT.Univariate analysis showed that there were significant differences in gender composition, type of transplant donor, number of natural killer cells and B lymphocytes in peripheral blood at 30 days after allo-HSCT, positive rate of plasma Epstein-Barr virus deoxyribonucleic acid (EBV-DNA) and incidence rate of acute graft-versus-host disease (aGVHD) between the 2 groups (all P<0.05).Multivariate Logistic regression analysis showed that female ( OR=3.196, 95% CI: 1.144-8.929), positive plasma EBV-DNA ( OR=17.523, 95% CI: 5.449-56.344) and aGVHD ( OR=3.156, 95% CI: 1.161-8.575) were independent risk factors for PTLD after allo-HSCT in TM children (all P<0.05).The ROC curve analysis showed that positive plasma EBV-DNA had an excellent accuracy in predicting the occurrence of PTLD after allo-HSCT (sensitivity was 0.796, specificity was 0.800, area under the curve was 0.803).If combined with aGVHD and gender, the area under the curve for the prediction of PTLD increased to 0.831. Conclusions:Female, positive plasma EBV-DNA and aGVHD are independent risk factors for PTLD after allo-HSCT in children with TM.It provides useful early warnings for the prediction and prevention of PTLD.