Objective:The components of Rubus parvifolius L. were analyzed based on UPLC-MS/MS technology and combined with network pharmacology analysis to explore the mechanism of action of Rubi Parvifolii Radix in treating inflammation, cough, fever, influenza and sore throat. Method:The chemical constituents of Rubi Parvifolii Radix were identified according to the information of mass spectrometry. The network pharmacology was used to analyze the corresponding targets and related pathways of its chemical components, and the "component-target-pathway" interaction diagram was drawn. PyMOL 2.5.7 software wasused to perform molecular docking between active components and key targets.Results:Twenty chemical components were identified by UPLC-MS/MS, and 15 components were screened out by network pharmacology, which can be used as quality markers of Rubi Parvifolii Radix, namely Azelaic acid, Procyanidol B3, Caprolactam, Bis (2-ethylhexyl) adipate, Cryptochlorogenic acid, 3-O-Feruloylquinic, Ellagic acid, Aurantiamide acetate, 2 α,3 β,19 α,23-Tetrahydroxyurs-12-en-28-oic acid, L-Epicatechin, (E)-3-Indoleacrylic acid, Euscaphic acid, Suberic acid, Diisononyl phthalate and Prodelphinidin T4. Molecular docking showed that 5 compounds compared with the reference substance could bind to the target proteins of disease well. Conclusions:The 15 active ingredients in Rubi Parvifolii Radix, including Caprolactam and (E)-3-Indoleacrylic acid, may play a therapeutic role in treating colds, high fever, sore throat, and inflammation by acting on targets such as AKT1 and TNF. This provides a certain reference for the clinical application of Rubi Parvifolii Radix.

Objective:To identify the components of Prunus mume f. viridicalyx based on ultra performance liquid chromatography-QE-Orbitrap mass spectrometry (UPLC-QE-Orbitrap-MS); To predict and analyze its substances and mechanisms to exert anti-depression effects combined with network pharmacology.Methods:UPLC-QE Orbitrap MS technology was used to analyze the chemical components of Prunus mume f. viridicalyx. Based on ChemSpider, mzCloud online platform, orbitrap TCM library and existing literature research, the secondary mass spectra of target compounds were compared and confirmed to identify the chemical composition of Prunus mume f. viridicalyx. The active components of the Prunus mume f. viridicalyx were screened. The Swiss Target Prediction database was used to predict targets with high correlation to active components in Prunus mume f. viridicalyx, and obtaining depression related disease targets from GeneCards and DisGeNET databases. The intersection targets of constituents and diseases were obtained using Venny platform. Protein-protein interaction network (PPI) was constructed by using String database, and the core targets were screened. Gene ontology function and Kyoto encyclopedia of genes and genomes pathway enrichment analysis of potential core targets were performed by using David database, and "active component-core target-signal pathway" network was constructed. PyMOL software was used to perform molecular docking between active components and key targets.Results:A total of 54 components, including organic acids, flavonoids and their glycosides, alkaloid, amino acids and other compounds were identified from Prunus mume f. viridicalyx. A total of 22 active components were screened and 92 active components and disease intersection targets were identified. A total of 13 core targets were screened through PPI network, including tumor necrosis factor, albumin, amyloid beta-protein precursor, AKT serine/threonine kinase 1 and so on. Enrichment analysis showed that Prunus mume f. viridicalyx mainly participated in transcription from RNA polymerase Ⅱ promoter, gene expression, protein binding and other functions, and presented the effects of anti-depression through MAPK, Toll-like receptor signaling pathway and other pathways. 12 key targets and 7 key active components were further obtained through the analysis of the "active component-core target-signal pathway" network, three of them were confirmed as kaempferol, quercetin, and isorhamnetin by reference substance. Molecular docking showed that 3 compounds could bind to the target proteins of depression well.Conclusion:Prunus mume f. viridicalyx exerts antidepressant effects through multiple components, targets, and pathways, mainly through the MAPK signaling pathway.

Doxorubicin(DOX)is a commonly administered chemotherapy drug for treating hematological malignancies and solid tumors;however,its clinical application is limited by significant cardiotoxicity.Cynaroside(Cyn)is a flavonoid glycoside distributed in honeysuckle,with confirmed potential biological functions in regulating inflammation,pyroptosis,and oxidative stress.Herein,the effects of Cyn were evaluated in a DOX-induced cardiotoxicity(DIC)mouse model,which was established by intraperitoneal injections of DOX(5 mg/kg)once a week for three weeks.The mice in the treatment group received dexrazoxane,MCC950,and Cyn every two days.Blood biochemistry,histopathology,immunohistochemistry,reverse transcription-quantitative polymerase chain reaction(RT-qPCR),and western blotting were conducted to investigate the cardioprotective effects and potential mechanisms of Cyn treatment.The results demonstrated the significant benefits of Cyn treatment in mitigating DIC;it could effectively alleviate oxidative stress to a certain extent,maintain the equilibrium of cell apoptosis,and enhance the cardiac function of mice.These effects were realized via regulating the transcription levels of pyroptosis-related genes,such as nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),caspase-1,and gasdermin D(GSDMD).Mechanistically,for DOX-induced myocardial injury,Cyn could significantly modulate the expression of pivotal genes,including adenosine monophosphate-activated protein kinase(AMPK),peroxisome proliferator-activated receptor γ coactivator-1α(PGC-1α),sirtuin 3(SIRT3),and nuclear factor erythroid 2-related factor 2(Nrf2).We attribute it to the mediation of AMPK/SIRT3/Nrf2 pathway,which plays a central role in preventing DOX-induced cardiomyocyte injury.In conclusion,the present study confirms the therapeutic potential of Cyn in DIC by regulating the AMPK/SIRT3/Nrf2 pathway.

ObjectiveTo study the effect of Qizhu Kang'ai prescription （QZAP） on the gluconeogenesis enzyme phosphoenolpyruvate carboxykinase 1 （PCK1） in the liver of mouse model of liver cancer induced by diethylnitrosamine （DEN） combined with carbon tetrachloride （CCl₄） and Huh7 cells of human liver cancer， so as to explore the mechanism on regulating metabolic reprogramming and inhibiting cell proliferation of liver cancer cells. MethodDEN combined with CCl₄ was used to construct a mouse model of liver cancer via intraperitoneal injection. A normal group， a model group， and a QZAP group were set up， in which QZAP （3.51 g·kg^-1） or an equal volume of normal saline was administered daily by gavage， respectively. Serum and liver samples were collected after eight weeks of intervention. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， γ-glutamyltransferase （γ-GT）， and alpha-fetoprotein （AFP） in mice were detected to evaluate liver function changes of mice in each group. Hematoxylin-eosin （HE） staining and Sirius red staining were used to observe pathological changes in liver tissue. In the cell experiment， Huh7 cells were divided into blank group， QZAP low， medium， and high dose groups and/or PCK1 inhibitor （SKF-34288 hydrochloride） group， and Sorafenib group. The corresponding drug-containing serum and drug treatment were given， respectively. Cell counting kit-8 （CCK-8） method， colony formation experiment， Edu fluorescent labeling detection， intracellular adenosine triphosphate （ATP） content detection， and cell cycle flow cytometry detection were used to evaluate the proliferation ability， energy metabolism changes， and change in the cell cycle of Huh7 cells in each group. Western blot was used to detect the protein expression levels of PCK1， serine/threonine kinase （Akt）， phosphorylated Akt （p-Akt）， and cell cycle-dependent protein kinase inhibitor 1A （p21）. ResultCompared with the model group， the pathological changes such as cell atypia， necrosis， and collagen fiber deposition in liver cancer tissue of mice in the QZAP group were alleviated， and the number of liver tumors was reduced （P<0.01）. The serum ALT， AST， γ-GT， and AFP levels were reduced （P<0.01）. At the cell level， compared with the blank group， low， medium， and high-dose groups of QZAP-containing serum and the Sorafenib group could significantly reduce the survival rate of Huh7 cells （P<0.01） and the number of positive cells with Edu labeling （P<0.01） and inhibit clonal proliferation ability （P<0.01）. The QZAP groups could also reduce the intracellular ATP content （P<0.05） and increase the distribution ratio of the G₀/G₁ phase of the cell cycle （P<0.05） in a dose-dependent manner. Compared with the model group and blank group， PCK1 and p21 protein levels of mouse liver cancer tissue and Huh7 cells in the QZAP groups were significantly reduced （P<0.05，P<0.01）， and the p-Akt protein level was significantly increased （P<0.01）. Compared with the blank group， the ATP content and cell survival rate of Huh7 cells in the SKF-34288 hydrochloride group were significantly increased （P<0.05）， but there was no statistical difference in the ratio of Edu-positive cells and the proportion of G₀/G₁ phase distribution. Compared with the SKF-34288 hydrochloride group， the QZAP combined with the SKF-34288 hydrochloride group significantly reduced the ATP content， cell survival rate， and Edu-positive cell ratio of Huh7 cells （P<0.05） and significantly increased the G₀/G₁ phase distribution proportion （P<0.05）. ConclusionQZAP may induce the metabolic reprogramming of liver cancer cells by activating PCK1 to promote Akt/p21-mediated tumor suppression， thereby exerting an anti-hepatocellular carcinoma proliferation mechanism.

Objective:To establish UPLC fingerprint method and 2 contents determination methods of Buddleja officinalis; To provide a reference for improving the quality control standard and evaluation of Buddleja officinalis from different habitats.Methods:UPLC method was used to establish the fingerprints of 17 batches of Buddleja officinalis. The similarity evaluation, clustering analysis, principal component analysis and orthogonal partial least squares discriminant analysis were used to compare the quality differences of Buddleja officinalis from different habitats. The contents of acteoside and linarin in Buddleja officinalis were determined.Results:There were 12 common peaks in UPLC fingerprints of Buddleja officinalis, six of which were identified as echinacoside, acteoside, cynaroside, isoacteoside, linarin, and apigenin. The fingerprint similarity of 17 batches of Buddleja officinalis was more than 0.9; Buddleja officinalis from different habitats were classified into 2 groups. Five differential markers were determined by OPLS-DA analysis. The order of significance was acteoside > peak 3 > echinacoside > isoacteoside > linarin. Edgeworthia chrysantha was identified by the method of fingerprint as counterfeit. The results of content determination showed that the content of Buddleja officinalis in Hubei and Sichuan was the high and stable.Conclusion:The method can effectively analyze the differences of Buddleja officinalis from different habitats, and provide reference for the quality control of Buddleja officinalis.

ObjectiveTo investigate the changes in the pathogen spectrum of viral diarrhea in local pediatric inpatients as well as any variations in genotypes of major pathogens during the COVID-19 control period. MethodsFecal samples were collected from the children <5 years who were hospitalized due to acute gastroenteritis in a pediatric hospital in Shanghai. PCR test was carried out to detect rotavirus， norovirus， sapovirus， astrovirus and enteric adenovirus， and then genotyping was performed for major pathogens. ResultsOut of 546 samples， 37.55% tested positive for virus with the following positive rate ranking： norovirus GⅡ （22.16%）， group A rotavirus （16.12%）， astrovirus （2.93%）， enteric adenovirus （2.38%）， sapovirus （0.92%） and norovirus GⅠ （0.18%）. The predominant genotype within norovirus GⅡ were GⅡ.4［P31］ and GⅡ.4［P16］ with a proportion of 24.79% and 14.05% respectively. The detection rate of GⅡ.4［P31］ dropped significantly over the 2-year period （χ²＝16.140，P<0.001）. In addition， an emerging rotavirus genotype G8P ［8］， which was rarely found nationally， was discovered for the first time locally with an increasing proportion， accounting for 7.95% of all rotavirus positive cases. Phylogenic analysis demonstrated that the representative strains of this genotype were genetically closer to the DS-1-like G8P ［8］ strain found in Southeast Asia. ConclusionThe changes in the prevalence of various norovirus genotypes together with the emergence of rare rotavirus genotype in the local area illustrate the importance of continuous monitoring of viral diarrhea and genotyping of key pathogens. Increased local activity of the rare genotype also adds new parameters in the efficacy evaluation of marketed vaccines and development of potential new vaccines in near future.

BACKGROUND:Numerous basic and clinical trials have confirmed that the low survival rate after bone marrow mesenchymal stem cell transplantation is a serious constraint on its long-term therapeutic effect.Previous studies have shown that apoptosis-related factors play an important role in the apoptosis of bone marrow mesenchymal stem cells,of which apoptosis-inducing factor may be a key factor. OBJECTIVE:Bone marrow mesenchymal stem cells,of which apoptosis-inducing factor was knocked down,were transplanted into infarcted myocardium of mice,aiming to certify the importance of apoptosis-inducing factor in the survival of bone marrow mesenchymal stem cells to further recover cardiac function after infarction. METHODS:Firstly,bone marrow mesenchymal stem cells were infected with LV-AIF-shRNA lentivirus to down-regulate the expression of apoptosis-inducing factor protein.Flow cytometry,western blot assay,and RT-qPCR were used to detect the infection efficiency of lentivirus.CCK-8 assay was used to detect the cell viability of bone marrow mesenchymal stem cells with apoptosis-inducing factor knockdown under hypoxic and ischemic conditions.Then,with the mouse model of acute myocardial infarction constructed,the normal bone marrow mesenchymal stem cells and bone marrow mesenchymal stem cells with apoptosis-inducing factor gene knockdown were transplanted into the model,respectively.The expression of apoptosis-inducing factor was examined by fluorescence immunoassay.Serum brain natriuretic peptide levels were detected by ELISA.Cardiac ultrasound was used to detect cardiac function.Myocardial fibrosis was observed by Masson staining.The expression of SRY gene was detected by RT-qPCR in apoptosis-inducing factor-knocked bone marrow mesenchymal stem cells after transplantation,reflecting cell survival. RESULTS AND CONCLUSION:(1)Bone marrow mesenchymal stem cells with apoptosis-inducing factor gene knockdown were successfully established by LV-AIF-shRNA lentivirus infection,following 97.7%of infection efficiency,and notably decline of the expression of apoptosis-inducing factor(P<0.001).(2)Under ischemia and hypoxia,the cell viability of apoptosis-inducing factor knockdown bone marrow mesenchymal stem cells was significantly increased compared with normal bone marrow mesenchymal stem cells.(3)Compared with normal bone marrow mesenchymal stem cells after transplantation,the survival number of bone marrow mesenchymal stem cells in the infarcted myocardium after apoptosis-inducing factor gene knockdown was significantly increased to 3.71 times(P<0.001),and the apoptosis-inducing factor protein expression and myocardial fibrosis degree in the infarcted area were significantly reduced.(4)Compared with normal bone marrow mesenchymal stem cells,the serum brain natriuretic peptide level of bone marrow stem cells with apoptosis-inducing factor gene knockdown after transplantation was significantly decreased(P<0.05),and left ventricular ejection fraction and left ventricular shortening fraction were significantly improved(P<0.05).(5)These findings confirm that apoptosis-inducing factor gene knockdown can reduce myocardial fibrosis and improve cardiac function after acute myocardial infarction via enhancing the bone marrow mesenchymal stem cell viability and increasing the bone marrow mesenchymal stem cell survival after transplantation in the donor.

Objective:To observe the effect of Wenyang Huazhuo recipe (WYHZ) on pregnancy outcomes in patients with unexplained recurrent spontaneous abortion (URSA).Methods:A retrospective cohort study was used and 240 cases of pregnancy URSA patients with spleen kidney yang deficiency syndrome at the Reproductive Center, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine and Department of Gynaecology and Obstetrics, General Hospital of Tianjin Medial University from November 2018 to March 2022 were recruited. The patients were assigned to WYHZ group (120 cases) and control group (120 cases) based on whether accepting oral WYHZ from 3 months before pregnancy to 8-12 weeks after pregnancy regularly. Pregnancy outcomes, perinatal period outcomes and obstetric complications were compared between the two groups. Multivariate logistic regression were used to analyze the effect of WYHZ on pregnancy outcome.Results:The live birth rate and the clinical pregnancy rate in WYHZ group [86.67% (104/120), 90.00% (108/120)] were higher than those in control group [71.67% (86/120), P=0.004; 75.00% (90/120), P=0.002]. The early abortion rate in WYHZ group [10.08% (12/119)] was lower than that in control group [25.86% (30/116), P=0.002], the differences were statistically significant. There were no significant differences in perinatal outcome and incidence rate of obstetric complications between the two groups (all P>0.05). Multivariate logistic regression analysis showed that the application of WYHZ significantly improved the live birth rate ( OR=8.818, 95% CI: 3.556-21.871, P<0.001) and sustained pregnancy rate ( OR=11.261, 95% CI: 4.262-29.751, P<0.001) in URSA patients. Conclusion:WYHZ using before and during pregnancy can significantly improve the live birth rate and the clinical pregnancy rate of URSA patients with syndrome of yang deficiency of spleen and kidney, and improve their pregnancy outcomes.

Objective:To observe the effect of Wenyang Huazhuo recipe (WYHZ) on pregnancy outcomes in patients with unexplained recurrent spontaneous abortion (URSA).Methods:A retrospective cohort study was used and 240 cases of pregnancy URSA patients with spleen kidney yang deficiency syndrome at the Reproductive Center, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine and Department of Gynaecology and Obstetrics, General Hospital of Tianjin Medial University from November 2018 to March 2022 were recruited. The patients were assigned to WYHZ group (120 cases) and control group (120 cases) based on whether accepting oral WYHZ from 3 months before pregnancy to 8-12 weeks after pregnancy regularly. Pregnancy outcomes, perinatal period outcomes and obstetric complications were compared between the two groups. Multivariate logistic regression were used to analyze the effect of WYHZ on pregnancy outcome.Results:The live birth rate and the clinical pregnancy rate in WYHZ group [86.67% (104/120), 90.00% (108/120)] were higher than those in control group [71.67% (86/120), P=0.004; 75.00% (90/120), P=0.002]. The early abortion rate in WYHZ group [10.08% (12/119)] was lower than that in control group [25.86% (30/116), P=0.002], the differences were statistically significant. There were no significant differences in perinatal outcome and incidence rate of obstetric complications between the two groups (all P>0.05). Multivariate logistic regression analysis showed that the application of WYHZ significantly improved the live birth rate ( OR=8.818, 95% CI: 3.556-21.871, P<0.001) and sustained pregnancy rate ( OR=11.261, 95% CI: 4.262-29.751, P<0.001) in URSA patients. Conclusion:WYHZ using before and during pregnancy can significantly improve the live birth rate and the clinical pregnancy rate of URSA patients with syndrome of yang deficiency of spleen and kidney, and improve their pregnancy outcomes.

Objective:To compare the surgical efficacy and perioperative complications of total intracorporeal neobladder (ICNB) and extracorporeal neobladder (ECNB) reconstruction during robot-assisted radical cystectomy.Methods:We retrospectively reviewed our single-institutional, prospectively collected database for consecutive patients who underwent RARC with an ileum neobladder from January 2017 to June 2022. A total of 265 patients were collected, 158 in ICNB group and 107 in ECNB group. The two groups were matched 1∶1 according to gender, age, BMI, ASA score, NAC, history of abdominal surgery and tumor characteristics using propensity score matching (PSM) method. After PSM, operating time, estimated blood loss and perioperative complications were compared between ICNB and ECNB group. A total of 186 patients were paired (93 patients in each group). There were 91 males and 2 females in ICNB group. The median age of ICNB group was 64(55, 68) years; median BMI was 23.54(22.39, 25.39) kg/m 2; There were 91 cases with Clinical stage ≤T 2 stage and 15 cases with ASA score ≥3; 25 cases received neoadjuvant chemotherapy. There were 91 males and 2 females in ECNB group. The median age of ECNB group was 63(52, 67) years; median BMI was 23.31(20.76, 24.80) kg/m 2; There were 90 cases with Clinical stage ≤T 2 stage and 8 cases with ASA score ≥3; 12 cases received neoadjuvant chemotherapy. In the ECNB group, after completing the radical cystectomy and pelvic lymph node dissection with robot assistance, the traditional lower abdominal incision was used to pull the intestinal tube outside the body to complete the preparation of the new bladder and then restore it to the pelvic cavity for anastomosing, while in the ICNB group, the reconstruction of the new bladder in situ was always completed in the body cavity with robot assistance. The operative time, intraoperative blood loss and perioperative complications were compared between the two groups. Results:The median operative time in ICNB and ECNB groups was 302(261, 350) min and 337(285, 397) min, respectively ( P=0.003); median intraoperative blood loss was 300(225, 500) ml and 500(300, 650) ml, respectively ( P<0.01); perioperative blood transfusion rates were 4.3%(4/93) and 14.0%(13/93), respectively ( P=0.022); all the differences between the 2 groups were statistically significant. 90-d overall complications rate was significant lower in ICNB group (38.7% vs. 61.3%, P=0.002). There was no difference in 90-d major complications (8.6% vs. 16.1%, P=0.119). On multivariate logistic regression analysis, operating time( OR=1.004, 95% CI 1.000-1.007, P=0.040), surgical approach (ICNB/ECNB)( OR=2.248, 95% CI 1.220-4.141, P=0.009) and age( OR=1.036, 95% CI 1.002-1.072, P=0.037) were associated with 90-day overall complications. Operating time( OR=1.036, 95% CI 1.002-1.072, P=0.037), neoadjuvant chemotherapy( OR=0.198, 95% CI 0.059-0.663, P=0.009) and ASA score( OR=0.167, 95% CI 0.044-0.634, P=0.009) were associated with 90-day high-grade complications. Conclusion:Intracorporeal neobladder during robot-assisted radical cystectomy is safe and feasible. Compared with ECNB, ICNB significantly improved perioperative outcomes such as shorter operating time, fewer estimated blood loss and perioperative complications.