Objective To investigate the effects of Sini Powder on serum hypothalamic-pituitary-adrenal(HPA)axis-related hormones and inflammatory factors in liver cancer mice with comorbid depression,and to evaluate its effect on depressive behavior and tumor proliferation activity.Methods Forty-eight specific pathogen-free female C57BL/6 mice were randomly assigned to either a blank(n=8)or model group(n=40).The modeling group was subjected to chronic unpredictable mild stress(CUMS)for six weeks.Both groups underwent orthotopically transplanted liver tumor surgery at the end of the fourth week of CUMS treatment.At the end of the sixth week of CUMS treatment,color Doppler ultrasonography was used to observe tumor formation in the orthotopic transplantation liver tumors,and the tail suspension test was used to assess depressive behavior.Non-tumor-bearing and deceased mice were excluded.The remaining model group mice were stratified by tail suspension immobility time and randomly assigned to the following groups:model group(distilled water),Fluoxetine group(5.0 mg/kg),and Sini Powder low-dose,medium-dose,and high-dose groups(5.2,10.4,and 20.8 g/kg,respectively),with six mice per group.The treatments were administered once daily for 21 consecutive days.After treatment,depressive behaviors were assessed using the open field,tail suspension,and forced swimming tests.The proliferation status of the orthotopic liver transplantation tumor was evaluated by measuring the size of the tumor,observing pathological changes in the tumor tissue through hematoxylin and eosin staining,and detecting the positive cell rate of proliferating cell nuclear antigen(Ki-67)in the tumor tissue using immunohistochemistry.The levels of HPA axis-related hormones in serum,such as corticotropin-releasing hormone(CRH),adrenocorticotropic hormone(ACTH),corticosterone(CORT),as well as inflammatory factors such as tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)were measured using an enzyme-linked immunosorbent assay.Western blotting was used to assess mitogen-activated protein kinase(MAPKs)phosphorylation and the expression of nuclear factor-κB(NF-κB),NOD-like receptor family pyrin domain-containing receptor 3(NLRP3),and cysteine aspartic protease-1(Caspase-1)in orthotopic tumors.Results Compared with the blank group,the model group showed reduced total distance traveled in open field test,prolonged immobility times in the tail suspension and forced swimming tests(P<0.05,P<0.01),indicating successful establishment of the liver cancer with comorbid depression mice model.Also,the model group showed increased orthotopic tumor volume(P<0.01),and elevated serum CRH,ACTH,CORT,TNF-α,IL-1β,and IL-6 levels(P<0.01).The phosphorylation of MAPKs in tumor tissues was suppressed(P<0.01),while NF-κB,NLRP3,and Caspase-1 expression levels were downregulated(P<0.01).Compared with the model group,Sini Powder medium-and high-dose groups exhibited increased total distance traveled in the open field test(P<0.05),reduced forced swimming test and prolonged total distance traveled in open field test(P<0.01),while Sini Powder high-dose group showed reduced immobility times in the tail suspension test(P<0.05).Also,Sini Powder low-dose,medium-dose,and high-dose groups showed slower tumor growth,histological changes,including vacuolization and necrosis,decreased Ki-67 positive cell rate(P<0.01),and reduced serum CRH,ACTH,CORT,TNF-α,IL-1β,and IL-6 levels(P<0.05).Additionally,the phosphorylation of MAPKs in tumor tissues was suppressed(P<0.01),and NF-κB,NLRP3,and caspase-1 expression levels were downregulated(P<0.01).Conclusion Sini Powder may alleviate depressive behaviors and suppress tumor proliferation activity in liver cancer mice with comorbid depression by modulating MAPKs activation,inhibiting NF-κB,NLRP3,and Caspase-1 expressions,and reducing serum inflammatory factors and HPA axis-related hormones levels.

Objective To investigate the effects of Sini Powder on serum hypothalamic-pituitary-adrenal(HPA)axis-related hormones and inflammatory factors in liver cancer mice with comorbid depression,and to evaluate its effect on depressive behavior and tumor proliferation activity.Methods Forty-eight specific pathogen-free female C57BL/6 mice were randomly assigned to either a blank(n=8)or model group(n=40).The modeling group was subjected to chronic unpredictable mild stress(CUMS)for six weeks.Both groups underwent orthotopically transplanted liver tumor surgery at the end of the fourth week of CUMS treatment.At the end of the sixth week of CUMS treatment,color Doppler ultrasonography was used to observe tumor formation in the orthotopic transplantation liver tumors,and the tail suspension test was used to assess depressive behavior.Non-tumor-bearing and deceased mice were excluded.The remaining model group mice were stratified by tail suspension immobility time and randomly assigned to the following groups:model group(distilled water),Fluoxetine group(5.0 mg/kg),and Sini Powder low-dose,medium-dose,and high-dose groups(5.2,10.4,and 20.8 g/kg,respectively),with six mice per group.The treatments were administered once daily for 21 consecutive days.After treatment,depressive behaviors were assessed using the open field,tail suspension,and forced swimming tests.The proliferation status of the orthotopic liver transplantation tumor was evaluated by measuring the size of the tumor,observing pathological changes in the tumor tissue through hematoxylin and eosin staining,and detecting the positive cell rate of proliferating cell nuclear antigen(Ki-67)in the tumor tissue using immunohistochemistry.The levels of HPA axis-related hormones in serum,such as corticotropin-releasing hormone(CRH),adrenocorticotropic hormone(ACTH),corticosterone(CORT),as well as inflammatory factors such as tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)were measured using an enzyme-linked immunosorbent assay.Western blotting was used to assess mitogen-activated protein kinase(MAPKs)phosphorylation and the expression of nuclear factor-κB(NF-κB),NOD-like receptor family pyrin domain-containing receptor 3(NLRP3),and cysteine aspartic protease-1(Caspase-1)in orthotopic tumors.Results Compared with the blank group,the model group showed reduced total distance traveled in open field test,prolonged immobility times in the tail suspension and forced swimming tests(P<0.05,P<0.01),indicating successful establishment of the liver cancer with comorbid depression mice model.Also,the model group showed increased orthotopic tumor volume(P<0.01),and elevated serum CRH,ACTH,CORT,TNF-α,IL-1β,and IL-6 levels(P<0.01).The phosphorylation of MAPKs in tumor tissues was suppressed(P<0.01),while NF-κB,NLRP3,and Caspase-1 expression levels were downregulated(P<0.01).Compared with the model group,Sini Powder medium-and high-dose groups exhibited increased total distance traveled in the open field test(P<0.05),reduced forced swimming test and prolonged total distance traveled in open field test(P<0.01),while Sini Powder high-dose group showed reduced immobility times in the tail suspension test(P<0.05).Also,Sini Powder low-dose,medium-dose,and high-dose groups showed slower tumor growth,histological changes,including vacuolization and necrosis,decreased Ki-67 positive cell rate(P<0.01),and reduced serum CRH,ACTH,CORT,TNF-α,IL-1β,and IL-6 levels(P<0.05).Additionally,the phosphorylation of MAPKs in tumor tissues was suppressed(P<0.01),and NF-κB,NLRP3,and caspase-1 expression levels were downregulated(P<0.01).Conclusion Sini Powder may alleviate depressive behaviors and suppress tumor proliferation activity in liver cancer mice with comorbid depression by modulating MAPKs activation,inhibiting NF-κB,NLRP3,and Caspase-1 expressions,and reducing serum inflammatory factors and HPA axis-related hormones levels.

With the development of molecular pharmacognosy, the advantages of DNA molecular markers in the identification of original plants of Chinese medicinal materials are becoming increasingly significant. To compensate for the limitations of existing markers in the quality supervision of Chinese medicinal materials, our team has independently designed a new molecular marker named DNA signature sequence(DSS). This marker is a nucleotide sequence that only appears in a specific taxonomic unit, with a length of 40 bp and high identification accuracy. This article aims to screen and verify the DSS markers that can accurately identify the original plants of the medicinal materials included in the volume one of the Chinese Pharmacopoeia, establish the operating procedure for developing standard nucleotide sequences, and lay a foundation for the widespread application of polymerase chain reaction in the quality supervision of traditional Chinese medicine. Firstly, the Chloroplast Genome Information Resource(CGIR) was searched for the chloroplast genome sequences of the test samples, species of the same genus, and common background species. IdenDSS was used to obtain the DSS tags and specific identification primers of the tested species. After DNA extraction, PCR amplification, sequencing, and sequence alignments, a total of 203 DSS markers of Chinese medicinal materials were obtained for validation. The above sequences were uploaded to the Traditional Chinese Medicine Molecular Identification Platform(www.herbsdna.com), and a standard DSS library was established for identifying the original plants of medicinal materials, serving as an important tool for quality supervision of Chinese materia medica. On this basis, an operating procedure for DSS development is formed, laying a foundation for further DSS screening and application based on more diverse genome sequences.
Plants, Medicinal/classification* ; DNA, Plant/genetics* ; Drugs, Chinese Herbal ; Pharmacopoeias as Topic ; Genetic Markers ; Medicine, Chinese Traditional

Child ; Humans ; Mpox (monkeypox)/prevention & control* ; Disease Outbreaks

ObjectiveTo investigate the effect of Shugan Quyu Jiedu prescription （SGQYJDF） on inducing ferroptosis in hepatocellular carcinoma cells based on the tumor protein 53 （p53）/solute carrier family 7 member 11 （SLC7A11）/glutathione peroxidase 4 （GPX4） pathway. MethodMHCC97H cells were divided into the blank serum group （10% blank serum medium）， SGQYJDF-containing serum low concentration group （5% SGQYJDF-containing serum and 5% blank serum medium）， SGQYJDF-containing serum medium concentration group （7.5% SGQYJDF-containing serum and 2.5% blank serum medium）， SGQYJDF-containing serum high concentration group （10% SGQYJDF-containing serum medium） and sorafenib group （sorafenib concentration of 10 μmol·L^-1 in 10% blank serum medium）. After 24 hours of intervention， the cell survival rate was detected by cell counting kit-8 （CCK-8） assay. The cell proliferation ability was detected by 5-ethynyl-2′-deoxyuridine （EdU） staining. The intracellular ferrous ion （Fe²⁺） level was detected by ferrous ion fluorescent probe （FerroOrange） staining. The intracellular malondialdehyde （MDA） and glutathione （GSH） levels were detected by colorimetric assays. The ultrastructure of mitochondria was observed by transmission electron microscopy. The expression levels of ferroptosis-related proteins p53， SLC7A11 and GPX4 were detected by Western blot. ResultIn terms of cell viability， compared with the blank serum group， the SGQYJDF group showed a dose-dependent decrease in the survival rate of MHCC97H cells. Effect of the medium and high concentrations of SGQYJDF on the survival rate of MHCC97H cells were significantly decreased （P<0.01）. Additionally， the results of the EdU assay showed that both the medium and high concentrations of SGQYJDF were able to inhibit the proliferation ability of MHCC97H cells （P<0.05， P<0.01）. Regarding the biochemical indicators of ferroptosis， compared to the blank serum group， the medium and high concentrations of SGQYJDF were able to dose-dependently increase the intracellular Fe²⁺ level （P<0.01）. The low， medium， and high concentrations of SGQYJDF were able to dose-dependently decrease the level of GSH in MHCC97H cells （P<0.01） and increase the level of MDA in the cells （P<0.05， P<0.01）. In terms of pathway-related protein expression， compared to the blank serum group， the medium and high concentrations of SGQYJDF could significantly increase the expression of p53 （P<0.01）. The low， medium， and high concentrations of SGQYJDF could significantly decrease the expression of GPX4 （P<0.01）. The high concentration of SGQYJDF could decrease the expression of SLC7A11 （P<0.01）. In terms of the cell morphology of ferroptosis， compared with the blank serum group， transmission electron microscopy revealed that the low concentration of SGQYJDF caused mitochondrial deformation， while the medium and high concentrations of SGQYJDF resulted in reduced mitochondrial volume， increased double-layer membrane density， and decreased mitochondrial cristae. These features were similar to those of sorafenib-induced ferroptosis. Furthermore， compared with the sorafenib group， the high concentration of SGQYJDF showed no statistically significant differences in cell survival rate， proliferation ability， Fe²⁺ level， MDA level， and GSH level. ConclusionThe results suggest that SGQYJDF may induce ferroptosis and inhibit proliferation in hepatocellular carcinoma MHCC97H cells by upregulating the expression of p53， suppressing the expressions of GPX4 and SLC7A11， downregulating the level of GSH， and leading to the accumulation of intracellular Fe²⁺ and MDA.

Objective:To study the alveolar bone remodeling of maxillary anterior teeth after extraction treatment and 2-year recovery period in adolescent patients with maxillary anterior arch protrusion.Methods:15 adolescent patients with maxillary anterior arch protru-sion were included,2 maxillary first premolars were extracted and implant anchorage combined with sliding method were used to close the extraction gap.CBCT images were taken before treatment(T0),after treatment(T1)and 2 years of recorvery period(T2),respectively.After multi-plane reconstruction with Dophin Imaging,the alveolar bone area(ABA)changes of maxillary central incisor,lateral incisor and canine at cementoenamel junction(CEJ-3 mm),root neck,central part,and root tip were measured and recorded as TAC,TA1,TA2 and TA3 respectively.The labial palatal alveolar crest to CEJ bone height(BH)of each tooth was recorded as BCL,BCP respectively.The data were analyzed by IBM SPSS statistics 25.0.Results:In T0-T2 phase,TA1 of each tooth was reduced.In T0-T1 phase,the horizontal adsorption of teeth was significantly correlated with ΔBCP,followed by ΔTA3.In T0-T2 phase,ΔBCP,ΔTA2,ΔTA3 and the horizontal adsorption of teeth showed low negative correlation.In T0-T1 phase,the vertical reduction of teeth was significantly positively correlated with ΔTAC,followed by low correlation with ΔTA3 and ΔTA1.Conclusion:In the treatment of anterior arch protrusion after extraction correction in adolescent patients the more the vertical reduction and horizontal adsorption of teeth in the treatment phase,the more the alveolar bone thickness and height around the tooth root in the maintenance phase,which were significantly positively correlated.Reasonable control of the vertical move-ment of teeth in the alveolar bone can improve the periodontal condition around the teeth to a certain extent.

Abstract OBJECTIVE To analyze the inflammation-related molecular targets of Pien Tze Huang in the treatment of hepatocellular carcinoma and to preliminary explore its mechanism. METHODS Obtain the ingredients and targets of Pien Tze Huang through TCMSP and BATMAN databases. Obtain the disease targets of hepatocellular carcinoma through Genecards, OMIM and TCGA databases. Take the intersection of compound targets and disease targets to get Pien Tze Huang’s target for the treatment of hepatocellular carcinoma. Obtain the related genes of inflammation pathway from the GSEA database, and then analyze the correlation between Pien Tze Huang’s therapeutic targets for hepatocellular carcinoma and inflammation-related genes to screen out inflammation-related targets, and explore the mechanism through GO and KEGG enrichment analysis. Then, single-factor cox analysis and LASSO regression were performed to construct related prognostic models. The 10 core targets were screened out through the protein-protein interaction(PPI) network. The model gene and the core target were intersected. The core compounds were screened out through the drug-compound-target network. Perform molecular docking verification between the core compound and the target. Construct a nomogram to assess the prognosis of patients. RESULTS Obtained 162 Pien Tze Huang targets, 522 hepatocellular carcinoma targets, 20 Pien Tze Huang therapeutic targets for hepatocellular carcinoma, and 16 inflammation-related targets. The enrichment analysis of GO and KEGG showed that their effects were mainly through biological functions such as monooxygenase activity, oxidoreductase activity, and chemical carcinogenesis-receptor activation. The ROC curve of the prognosis model calculated AUC as 0.780 in 1 year, 0.688 in 3 years, and 0.642 in 5 years, indicating that the model was reliable. The prognostic model intersects with the core target of PPI to get 5 targets: PON1, IGF2, NQO1, CCNB1 and IGFBP3. The nomogram was constructed using CCNB1, NQO1, and T staging, and its c-index was 0.726, indicating the reliability of the model. The drug-compound-target network suggested that quercetin was the core compound and targets the above two genes. CONCLUSION Pien Tze Huang’s treatment of hepatocellular carcinoma mainly uses quercetin to target CCNB1 and NQO1 to exert anti-inflammatory effects, and its prognostic model can be used to predict the survival of patients.

AIM:To investigate the inhibitory effect of the"decoction for soothing liver and removing stasis and toxicity(SGQYJDF)"on hepatocellular carcinoma(HCC)proliferation in nude mice by inducing ferroptosis via the tumor protein 53(p53)/solute carrier family 7 member 11(SLC7A11/xCT)/glutathione peroxidase 4(GPX4)pathway.METHODS:An ectopic subcutaneous tumor model was established by injecting SK-Hep-1 cells subcutaneously into the right axilla of nude mice.Upon formation of tumor,the mice were randomly divided into five groups(i.e.,control group,low-,medium-and high-dose SGQYJDF groups and medium-dose SGQYJDF plus Sorafenib group).Each group of mice was orally administered with the corresponding therapy for 14 consecutive days,during which the tumor size was observed regularly.At the end of treatment,the tumor growth inhibition rate was calculated based on tumor mass,and histopatho-logical changes were observed by HE staining.Then,the levels of malondialdehyde(MDA),glutathione(GSH)and fer-rous ions(Fe2+)were detected by colorimetric assays.The expression of the proliferation markers Ki-67 and GPX4 was de-tected by immunohistochemistry(IHC).The expression of p53 and xCT was detected by Immunofluorescence(IF).And the expression of p53,xCT and GPX4 was determined by Western blot.RESULTS:(1)SGQYJDF was found to dose-de-pendently decrease tumor volume(P＜0.01)and inhibit tumor mass growth(P＜0.01),and meanwhile,reduce the per-centage of Ki-67-positive cells(P＜0.01)and their proliferation ability in tumor tissues,as compared to the control group.(2)In terms of Ferroptosis-related indicators,SGQYJDF was found to dose-dependently increase the levels of Fe2+ and MDA but decrease the level of GSH in tumor tissues(P＜0.01),as compared to the control group.(3)In terms of protein expression,SGQYJDF was found to dose-dependently upregulate the expression of p53(P＜0.05)but inhibit the expres-sion of xCT(P＜0.05)and GPX4(P＜0.01).Notably,medium-dose SGQYJDF plus sorafenib showed a stronger effect than high-dose SGQYJDF.CONCLUSION:Our findings suggest that SGQYJDF can induce Ferroptosis to inhibit the proliferation of subcutaneously transplanted tumor tissues in nude mice by upregulating the expression of p53,and inhibit-ing the expression of xCT and GPX4.

Objective To observe the morphological changes in the testes and ovaries of adult 12th-generation Oncomelania hupensis bred for 12 winters in Weishan Lake areas. Methods The offspring of the adult O. hupensis snails bred in the Weishan Lake that were originated from the Yangzhou section of the Yangtze River was defined as the experiment group, while uninfected, adult O. hupensis snails captured from the marshland of the Yangzhou section of the Yangtze River served as the control group. Snails were dissected and intact testicular and ovarian specimens were sampled, routinely fixed, dehydrated, embedded, polymerized in an oven and sliced on an ultramicrotome. The sections were visualized under a transmission electron microscope, and the ultrastructure of the snail gonad was compared between the experiment and control groups. Results Transmission electron microscopy showed “9 + 2” microtubules on the transverse sections of the tails of sperm cells in the testes of male snails in the control group, with triangular acrosomes and spiral, dense nuclei seen in the tip, while in the experiment group, the “9 + 2” microtubules disappeared on the transverse sections of the tails of sperm cells in the testes of male snails, with low chromatin density found in the tip. Transmission electron microscopy revealed clear nucleolus and nuclear membranes in the ova of female snail ovaries, and displayed yolk body, liposomes and endoplasmic reticulum in the cytoplasm, bilayer twists of nuclear membrane and a uniform nucleolus in the control group, while in the experiment group, smooth nuclear membrane and unclear nucleolus were observed in the ova of female snail ovaries, with few contents seen within cells. Conclusions Following breeding for 12 winters in the Weishan Lake, the 12th-generation O. hupensis snails fail to fully adapt to the natural environment in northern China, and the remarkable changes in the ultrastructure of the snail gonad may be a cause of gradual decline and even extinction of O. hupensis in the Weishan Lake areas.