This study explores the therapeutic differences and mechanisms of salt-processed Phellodendri Chinensis Cortex in improving insulin resistance(IR) based on network pharmacology, molecular docking, and cellular experiments. The components and intersection targets of Phellodendri Chinensis Cortex in improving IR were collected from databases, and a "drug-component-target-disease" network and protein-protein interaction(PPI) network were constructed to screen core components and targets. A total of 29 active components and 240 intersection targets were identified, of which 13 were core targets. Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses were used to identify key signaling pathways, and molecular docking was performed to validate the binding activity between core components and targets. An IR model in HepG2 cells was induced using insulin combined with high glucose, and the effects of Phellodendri Chinensis Cortex before and after salt-processing on cell glucose consumption were evaluated. The expression of proteins related to the mitogen-activated protein kinase(MAPK) and phosphatidylinositol 3-kinase(PI3K)-protein kinase B(AKT) signaling pathways was detected by Western blot. The cellular experimental results showed that, compared with the model group, glucose consumption in the drug-treated groups was significantly increased(P<0.01), the phosphorylation level of extracellular regulated protein kinase(ERK) was decreased(P<0.05), the phosphorylation levels of PI3K and AKT were increased, and the expression of glucose transporter 4(GLUT4) was also upregulated(P<0.05). Furthermore, the effect of salt-processed Phellodendri Chinensis Cortex was better than that of raw Phellodendri Chinensis Cortex. The study demonstrates that Phellodendri Chinensis Cortex, both before and after salt-processing, improves IR by regulating the expression of related proteins in the MAPK and PI3K-AKT signaling pathways, with enhanced effects after salt-processing.
Humans ; Network Pharmacology ; Phellodendron/chemistry* ; Insulin Resistance ; Drugs, Chinese Herbal/chemistry* ; Hep G2 Cells ; Signal Transduction/drug effects* ; Molecular Docking Simulation ; Protein Interaction Maps/drug effects* ; Proto-Oncogene Proteins c-akt/genetics* ; Phosphatidylinositol 3-Kinases/genetics* ; Glucose/metabolism*

Medical institutions, with their clinical practice foundation and abundant human use experience data, have become important carriers for the inheritance and innovation of traditional Chinese medicine(TCM) and the "cradles" of the preparation of new TCM. To effectively promote the transformation of new TCM originating from the TCM clinical practice in medical institutions and establish an effective evaluation index system for the transformation of new TCM conforming to the characteristics of TCM, consensus experts adopted the literature research, questionnaire survey, Delphi method, etc. By focusing on the policy and technical evaluation of new TCM originating from the TCM clinical practice in medical institutions, a comprehensive evaluation from the dimensions of drug safety, efficacy, feasibility, and characteristic advantages was conducted, thus forming a comprehensive evaluation system with four primary indicators and 37 secondary indicators. The expert consensus reached aims to encourage medical institutions at all levels to continuously improve the high-quality research and development and transformation of new TCM originating from the TCM clinical practice in medical institutions and targeted at clinical needs, so as to provide a decision-making basis for the preparation, selection, cultivation, and transformation of new TCM for medical institutions, improve the development efficiency of new TCM, and precisely respond to the public medication needs.
Medicine, Chinese Traditional/standards* ; Humans ; Consensus ; Drugs, Chinese Herbal/therapeutic use* ; Surveys and Questionnaires

Abnormal amino acid metabolism promotes tumor progression by inducing malignant behaviors in tumor cells and altering the immune landscape within the tumor microenvironment. However, the underlying mechanisms remain unclear. In this study, we constructed colorectal cancer (CRC) organoids and patient-derived tumor xenograft (PDX) models, performing multifaceted validation to confirm that T-complex protein 1 subunit epsilon (CCT5), mediates the biosynthesis of aspartate and enhances sensitivity to anti-PD-L1 immunotherapy. Mechanistically, CCT5 directly binds to asparagine synthetase (ASNS) and promotes the synthesis of aspartate (Asn). The Asn-mTORC1 axis facilitates tumor cell proliferation while upregulating PD-L1 expression, which leads to a reduction in the number of effector CD8⁺ T cells. Treatment with l-asparaginase (ASNase) combined with anti-PD-L1 therapy effectively reverses the growth of CRC characterized by high CCT5 expression. In summary, we identify CCT5 as a potential biomarker to guide the combined use of ASNase and anti-PD-L1 antibodies in CRC treatment.

Objective To investigate the effects of medullary differentiation-2(MD-2)in obesity-induced myocardial injury by regulating adenosine 5'-monophosphate-activated protein kinase(AMPK)activity.Methods The obese mice model was established by feeding high-fat feed.The wild-type and MD-2 gene knockout mice were randomly divided into wild-type(WT)-control group(fed normally),WT-model group(fed with high-fat diet),knockout(KO)-control group(fed normally)and KO-model group(fed with high-fat diet)with 8 mice in each group.At week 16,creatine kinase-MB(CK-MB)and lactate dehydrogenase(LDH)were measured by reagent kits;the left ventricular ejection fraction(LVEF)and left ventricular fraction shortening(LVFS)were measured with ultrasonic apparatus;the pathological changes in myocardial tissue were observed by hematoxylin and eosin(HE)staining and Masson staining;the protein expression of phosphorylation AMPK(p-AMPK)was measured by Western blot.Results HE staining and Masson staining results revealed significant necrosis and fibrosis of the myocardial tissue in WT-model group,while the degrees of necrosis and fibrosis in KO-model group were significantly reduced.The LVEF values of the WT-control group,WT-model group,KO-control group and KO-model group were(83.98±7.58)％,(52.03±4.91)％,(76.42±3.89)％and(73.71±4.22)％,respectively;the LVFS values of each group were(53.61±8.51)％,(24.56±3.76)％,(48.14±5.00)％and(44.07±3.74)％,respectively;the CK-MB values of each group were(49.83±15.49),(83.75±11.90),(54.03±18.66)and(66.85±12.98)U·L-1,respectively;the LDH values of each group were(12.24±3.20),(27.90±6.10),(13.55±3.55)and(15.53±2.58)U·L-1,respectively;the relative expression levels of p-AMPK in each group were 221.31±88.76,46.29±10.07,148.66±32.02 and 161.49±78.11.Compared with WT-control group,the above indicators in WT-model group,compared with WT-model group,the above indicators in KO-model group,the differences were statistically significant(all P＜0.05).Conclusion MD-2 mediates high-fat feed induced myocardial injury in obese mice by inhibiting AMPK activity.

Objective To explore the protective mechanism of icariin on neuronal oxidative damage,providing a basic pharmacological basis for the treatment of cognitive impairment.Methods Glutamate was used to induce oxidative stress injury in HT22 cells.HT22 cells were divided into control group(normal cultured cells),model group(glutamate injury model)and experimental-L,-M,-H groups(5,10 and 20 μmol·L-1 icariin pretreatment for modeling,respectively).Cell proliferation was detected by cell counting kit-8(CCK-8)method;cytotoxicity was detected by lactate dehydrogenase(LDH)method;reactive oxygen species(ROS)levels were detected by flow cytometry;superoxide dismutase(SOD)levels were detected by biochemical kits;the expression levels of Kelch-like epichlorohydrin-related protein-1(Keap1),nuclear factor E2-related factor 2(Nrf2)were detected by Western blotting;the corresponding mRNA expression was detected by real-time fluorescence quantification polymerose chain reaction.Results The cell viability of control group,model group and experimental-L,-M,-H groups were(100.00±1.31)％,(66.38±2.44)％,(72.07±4.95)％,(82.41±3.57)％and(87.97±4.98)％;LDH release were(0.48±0.52)％,(18.82±2.09)％,(15.32±1.17)％,(10.37±1.39)％and(6.51±0.87)％;ROS level were(14.23±1.13)％,(41.74±1.60)％,(35.69±1.08)％,(33.28±1.69)％and(30.32±2.03)％;SOD levels were(54.84±1.17),(37.95±1.13),(48.02±1.28),(50.56±1.34)and(52.55±1.04)U·mg-1;Keap1 protein levels were 0.36±0.01,0.52±0.03,0.46±0.04,0.39±0.09 and 0.35±0.12;Nrf2 protein levels were 0.29±0.02,0.13±0.08,0.18±0.03,0.21±0.11 and 0.26±0.04;catalase(CAT)mRNA levels were 1.01±0.08,0.81±0.06,0.90±0.04,1.05±0.15 and 1.33±0.26;SOD mRNA levels were 1.09±0.12,0.83±0.03,0.86±0.08,0.94±0.08 and 1.09±0.16.Among the above indicators,the differences between the model group and the control group were statistically significant(all P＜0.01);the differences between the experimental-M,-H groups and the model group were statistically significant(P＜0.01,P＜0.05).Conclusion Icariin may activate the Keap1/Nrf2/antioxidant response element(ARE)signaling pathway,regulate the expression of related proteins,and reduce the level of ROS to effectively alleviate oxidative stress injury in neuronal cells.

Objectives:To compare the HbA 1c results on capillary electrophoresis (CE), cation exchange-high performance liquid chromatography (CE-HPLC), and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Methods:902 normal samples without hemoglobin variants and 83 samples with hemoglobin variants were collected in 2020. The variants were divided into α-chain, β-chain, γ-chain, and δ-chain variants. The 902 samples without hemoglobin variants were measured by CE, CE-HPLC, and MALDI-TOF MS. Three methods were used for measuring α-chain, β-chain, and γ-chain compared with the liquid chromatography-tandem mass spectrometry (LC-MS/MS) reference method. According to the EP9-A3 guideline, Passing-Bablok regression and concordance correlation coefficient (CCC) were performed for correlation and consistency, respectively. The imprecision was assessed by coefficient of variance (CV). The mean relative bias was calculated and compared with the lowest biological variation bias of 2.3%.Results:All three methods met the acceptance of imprecision requirements (<2%). The R 2 and CCC for the normal samples were all above 0.95 between the pairwise comparison of CE, CE-HPLC, and MALDI-TOF MS. The mean relative bias between MALDI-TOF MS and CE-HPLC was higher than the lowest biological variation bias of 2.3%. The R 2 results between CE and LC-MS/MS results of α-chain, β-chain, γ-chain variants were>0.95, and the mean relative biases for γ-chain variants exceeded the lowest biological variation bias of 2.3%. A bad correlation ( R2=0.66) for β-chain variants was shown between CE-HPLC and LC-MS/MS and the mean relative biases of all variant samples exceeded 2.3%. MALDI-TOF MS showed good correlations with LC-MS/MS for three groups, and the mean relative biases for γ-chain variants were higher than 2.3%. Conclusion:MALDI-TOF MS and CE-HPLC with CE had good comparability in the measurement of HbA 1c in normal samples, butthe three methods showed interferences from different types of Hb variants and the CE method was affected with less interference.

Objective:To explore the relationship between heart rate variability, severity, and prognosis in patients with acute cerebral infarction (ACI).Methods:A prospective study was conducted on 80 patients with acute cerebral infarction (ACI) admitted to Qinhuangdao Workers′ Hospital from February 2021 to March 2023. The severity of ACI patients was evaluated based on their National Institutes of Health Stroke Scale (NIHSS) scores, and they were divided into mild group (26 cases, NIHSS scores 0-15), moderate group (32 cases, NIHSS scores 16-30), and severe group (22 cases, NIHSS scores 31-45); And 30 healthy individuals who underwent physical examinations during the same period were selected as the control group. The improved Rankin Scale (MRS) was used to evaluate the prognosis of patients, who were divided into a good prognosis group (59 cases) and a poor prognosis group (21 cases) based on the scoring results. All patients underwent dynamic electrocardiogram examination, and the heart rate variability parameters of each group were compared [standard deviation of R-R interval for all sinus beats at 24 hours (SDNN), mean standard deviation of R-R interval for 5 consecutive minutes at 24 hours (SDNN Index), standard deviation of R-R interval mean every 5 minutes (SDANN), root mean square of all adjacent R-R interval differences (rMSSD), and the proportion of adjacent NN interval differences greater than 50 ms (PNN50%)]. The predictive value of heart rate variability parameters on the severity and prognosis of ACI patients was analyzed using receiver operating characteristic (ROC) curves. Logistic regression analysis was used to identify the risk factors for the severity and prognosis of ACI patients.Results:The SDNN, SDANN, SDNN Index, rMSSD, and PNN50% of the mild, moderate, and severe groups were significantly lower than those of the control group ( F=59.382, 11.859, 12.376, 11.699, 47.703, all P<0.01), and the severe group was less than the moderate group and less than the mild group (all P<0.05). The SDNN, SDANN, SDNN Index, rMSSD, and PNN50% in the poor prognosis group were lower than those in the good prognosis group ( t=4.536, 4.181, 5.091, 4.384, 2.851, all P<0.01). The results of logistic regression analysis showed that heart rate variability parameter was an independent influencing factor for the severity and poor prognosis of ACI patients (all P<0.01). The area under the curve for the combined diagnosis of the severity of ACI patients using SDNN, SDANN, SDNN Index, rMSSD, and PNN50% was 0.950, and the area under the curve for the combined prediction of ACI patients′ prognosis was 0.970. Conclusions:The heart rate variability parameter is an independent influencing factor on the severity and prognosis of ACI patients, and its predictive power for the severity and prognosis of ACI patients is high. It can be used as an effective indicator for evaluating the severity and prognosis of ACI patients.

Objective:To prepare mesenchymal stem cells with antioxidant capacity (AO-MSC ) from human umbilical cords and evaluate its cell biological properties.Methods:In control group,mesenchymal stem cells (MSC) were isolated by digesting human umbilical cord Wharton's Jelly tissues with 0.2％ collagenase Ⅱ,and the released cells were collected and cultured in an animal serum-free culture medium.In AO-MSC group,incompletely collagenase Ⅱ-digested tissue debris were allowed to adhere to flusk flat bottoms and the AO-MSC was harvested by adherent culture. The conventional digestion and culture method was used as control.MSC colony forming ability was evaluated by fibroblast colony forming assay (CFU-F).MSC proliferative capacity was evaluated by CCK-8 assay.The MSC surface markers were detected by using flow cytometry and immunofluorescence staining.The adipogenic and osteogenic capacity of MSC was evaluated by multi-differentiation in vitro,and the mRNA expression of genes that control adipogenic and osteogenic differentiation was detected by real-time fluorescence quantitative PCR (RT-qPCR );Moreover,the mRNA expression of antioxidant substances such as SOD-1,GSH,GAT,and NQO1 in MSC was also evaluated by RT-qPCR.Results:The AO-MSC isolated by this strategy reached a confluence of 80％-90％ at around 18 days and grew in a swirling pattern.Flow cytometry and immunofluorescence staining assays showed that CD73,CD29,CD105,CD90 were highly expressed and CD31,CD45,HLA-DR were scarcely expressed in AO-MSC.AO-MSC exhibited stronger self-renewal and differentiation ability compared to MSC.However,the in vitro adipogenic-osteogenic capacity of MSC in the control group was stronger than that of AO-MSC.RT-qPCR assay showed that AO-MSC expressed higher mRNA levels of antioxidant substances compared to MSC.Conclusion:Human AO-MSC is successfully prepared from human umbilical cord without animal serum.

Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75％(27/36);boys accounted for 64％(23/36).Among the 36 neonates,31(86％)had disease onset within 3 days after birth,and 19(53％)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61％)had left cerebral infarction and 13(36％)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75％)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72％)and seizures in 10 neonates(34％).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33％,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44％(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.

Objective To observe the intervention effect and mechanism of Chinese herbal compound(Zhenyuan Granules)on overweight in mice caused by estrogen deficiency.Methods An ovariectomized female mouse model was established to observe the effects of intragastric 14 week administration of Zhenyuan Granules on body mass and fat accumulation in mice.The 3T3-L1 mouse preadipocyte model was constructed to observe the effect of Zhenyuan Granules intervention on adipogenic differentiation.The expressions of key genes/proteins regulating adipocyte formation and fat synthesis in mouse adipose tissue and 3T3-L1 cells was detected to explore the mechanism of lipid-lowering effect of Zhenyuan Granules in vivo and in vitro.Results Zhenyuan Granules significantly decreased the body mass gain(P＜0.01),perigonadal and inguinal fat indexes(P＜0.05),significantly inhibited the differentiation of 3T3-L1 preadipocytes into mature adipocytes(P＜0.01),significantly downregulated the expression levels of transcription factors peroxisome proliferators activated receptor γ(PPARγ)and sterol-regulatory element binding protein 1c(SREBP-1c)and enzymes acetyl-CoA carboxylase 1(ACC1)and fatty acid synthase(FAS)in adipose tissue and 3T3-L1 preadipocytes(P＜0.05),significantly enhanced the phosphorylation of AMP-activated protein kinase(AMPK)and ACC1(P＜0.05).Conclusion Zhenyuan Granules can inhibit the formation of adipocytes and improve the overweight of female mice caused by estrogen deficiency.The mechanism is related to the activation of AMPK signaling pathway.It is suggested that Bushen Jianpi Chinese herbal compound Zhenyuan Granules is helpful for body mass control in postmenopausal women.