This article systematically analyzes the historical evolution of the name， origin， quality evaluation， harvesting， processing and other aspects of Picrorhizae Rhizoma by referring to the medical books， prescription books， and other documents of the past dynasties， combined with relevant modern research materials， in order to provide a basis for the development and utilization of famous classical formulas containing this medicinal herb. The research results indicate that Picrorhizae Rhizoma was first recorded in New Revised Materia Medica from the Tang dynasty. Throughout history， Huhuanglian has been used as its official name， and there are also aliases such as Gehu Luze， Jiahuanglian and Hulian. The main source of past dynasties is the the rhizomes of Picrorhiza kurrooa and P. scrophulariiflora. In ancient times， Picrorhizae Rhizoma was mainly imported by foreign traders via Guangzhou and other regions， and also produced in China， mainly in Xizang. In ancient times， it was harvested and dried in early August of the lunar calendar， while in modern times， it is mostly harvested from July to September， with the best quality being those with thick and crispy rhizomes without impurities， and bitter taste. Throughout history， Picrorhizae Rhizoma was collected， washed， sliced， and dried before being used as a raw material for medicine， it has a bitter and cold taste， mainly used to treat bone steaming， hot flashes， infantile chancre fever， and dysentery. There is no significant difference in taste and efficacy between ancient and modern times. Based on the research results， it is recommended that the rhizomes of P. scrophulariiflora in the 2020 edition of Chinese Pharmacopoeia， or the rhizomes of P. kurrooa， can be used in famous classical formulas containing this medicinal herb， which can be processed according to the processing requirements marked by the original formula. For those without clear processing requirements， the dried raw products are used as medicine.

Sepsis is a condition characterized by organ dysfunction resulting from the systemic inflammatory response triggered by an infection. Excessive inflammation and immunosuppression are intertwined, and severe cases may even develop into multiple organ failure. Studies have shown that indoleamine 2,3-dioxygenase 1-mediated tryptophan metabolism is involved in the occurrence and development of sepsis, and elevated plasma kynurenine levels and Kyn/Trp ratios are early indicators of sepsis development. In this paper, we provide a comprehensive summary of the role of IDO1 in the acute inflammatory phase of sepsis, late immunosuppression, and organ damage. This includes its regulation of inflammatory state, immune cell function, blood pressure, and other aspects. Additionally, we analyze preclinical studies on targeted IDO1 drugs. An in-depth understanding and study of IDO may help to understand the pathogenesis and clinical significance of sepsis and multiple organ damage from a new perspective and provide new research ideas for exploring its prevention and treatment methods.

OBJECTIVE To investigate the effect and mechanism of miR-152-3p on the resistance to paclitaxel(PTX)of PTX-resistant ovarian cancer cells(A2780T cells).METHODS ① Ovarian cancer parent cells(A2780 cells)and A2780T cells were treated with PTX(1.875,3.75,7.5,17 and 23 μmol·L-1)for 48 h.Cell viability was evaluated by MTT assay,and the 50%inhibitory concentration(IC50)and drug resistance index of A2780T cells were calculated.Western blotting was used to detect the expres-sions of resistance protein P-glycoprotein(P-gp),multidrug resistance related protein 1(MRP1)and adenosine triphosphate binding transporter G superfamily member 2(ABCG2).② Real-time fluorescent quantita-tive PCR(RT-qPCR)was used to detect the expressions of miR-152-3p in A2780 and A2780T cells.The lipid-mediated transient transfection technique was employed to transfect the miR-152-3p inhibitor to reduce miR-152-3p expression in A2780T cells(miR-152-3p inhibitor group),while the negative control(miR-152-3p NC)group was established.RT-qPCR was used to detect transfection efficiency,and the MTT method,scratch experiment,and flow cytometry were used to investigate the effects of the trans-fecting miR-152-3p inhibitor on survival,migration and apoptosis of A2780T cells.Western blotting was used to detect the protein expressions of Bax and Bcl-2 in A2780T cells.③ Bioinformatics analysis of databases including miRDB,Targetscan,miRWalk,and Starbase predicted the target genes of miR-152-3p that were verified by Western blotting to detect the protein expression of PTEN in A2780T cells of the miR-152-3p inhibitor and miR-152-3p NC groups,and RT-qPCR to detect the PTEN mRNA expression in A2780 and A2780T cells.Then,the lipid-mediated transient transfection technique was used to transfect PTEN siRNA to silence PTEN expression in A2780T cells(PTEN siRNA group).The siRNA negative control(siRNA NC)group was established.RT-qPCR was used to detect transfection efficiency,the MTT method was employed to measure the survival rate and IC50 value,and Western blotting was used to assess the protein expressions of P-gp,MRP1,and ABCG2 in A2780T cells after silencing PTEN expression.RESULTS ①After treatment with PTX,the cell survival rates were decreased in A2780 and A2780T cells(P<0.05),and the resistance index of A2780T cells was 2.8.Compared with A2780 cells,the protein expressions of P-gp and MRP1 and ABCG2 were highly expressed in A2780T cells(P<0.05,P<0.01).② RT-qPCR showed that the expression of miR-152-3p in A2780T cells was higher than that of A2780 cells(P<0.01).Compared with the miR-152-3p NC group,A2780T cell viability(P<0.05,P<0.01)and cell migration capability(P<0.05)were significantly inhibited,while the apoptosis rate increased(P<0.01)in miR-152-3p inhibitor group.Moreover,the protein expression of Bax was increased(P<0.01),but Bcl-2 decreased(P<0.05).③ Bioinformatics analysis suggested that PTEN was a target gene of the miR-152-3p,and the verified results showed that the PTEN protein expression in A2780T cells of the miR-152-3p inhibitor group was lower than that of the miR-152-3p NC group(P<0.05),and PTEN mRNA expression in A2780T cells was higher than that in A2780 cells(P<0.01).After silencing the expression of PTEN in A2780T cells,the cell viability was significantly reduced(P<0.05,P<0.01),while the IC50 value was reduced(P<0.01)compared with the siRNA NC group.In addition,the protein expressions of P-gp,MRP1 and ABCG2 were decreased(P<0.05,P<0.01).CONCLUSION miR-152-3p is highly expressed in A2780T cells,and down-regulation of its expression may inhibit proliferation and migration,prompt apoptosis and reduce the resistance to PTX of A2780T cells,which is made possible by inhibiting expression of its target gene PTEN.

Nitazoxanide is an FDA-approved antiprotozoal drug. Our previous study found that oral administration of nitazoxanide inhibited Western diet (WD)-induced hepatic steatosis in ApoE^-/- mice. However, the specific mechanism remains to be elucidated. In the present study, we performed an untargeted metabolomics approach to reveal the effect of nitazoxanide on the liver metabolic profiles in WD-fed ApoE^-/- mice, and carried out the cellular experiments to elucidate the underlying mechanisms. UPLC-MS-based untargeted metabolomics analysis was used to investigate the effect of nitazoxanide on global metabolite changes in liver tissues. The differential metabolites were screened for enrichment analysis and pathway analysis. Hepatocytes were treated with tizoxanide, the metabolite of nitazoxanide, to investigate the underlying mechanism based on the findings in metabolomics study. The improvement of liver lipid metabolism disorders by nitazoxanide treatment in WD-fed ApoE^-/- mice was mainly through regulating glycerophospholipid metabolism, D-glutamine and glutamate metabolism, glutathione metabolism, and arginine biosynthesis metabolism. Tizoxanide, the active metabolite of nitazoxanide, increased glutathione (GSH) contents and glutamate-cysteine ligase catalytic subunit (Gcl-c) and glutathione reductase (Gsr) mRNA expressions in HepG2 cells. Tizoxanide increased cystathionine β-synthase (CBS) and phosphatidylethanolamine N-methyltransferase (PEMT) protein levels, inhibited lipid accumulation in hepatocytes induced by free fatty acid (FFA). Tizoxanide increased S-adenosyl-L-homocysteine hydrolase (SAHH) protein levels in HepG2 cells and mouse primary liver cells stimulated with free fatty acid (FFA). Tizoxanide increased N-acetyl glutamate synthase (Nags) and carbamoylphosphate synthetase 1 (Cps1) mRNA expressions in HepG2 cells. In conclusion, nitazoxanide improves WD-induced hepatic steatosis in ApoE^-/- mice and the underlying mechanisms include increasing CBS expression, GSH content, PEMT protein expression, Nags and Cps1 mRNA expression in hepatocytes.

Chronic prostatitis is a process of kidney deficiency and blood stasis mixed with various pathological factors involving the essence chamber,which is manifested as kidney deficiency and blood stasis.Based on the concept of the"brain-heart-kidney-es-sence chamber"axis of medication,Xiongji Formula is applied to the treatment of chronic prostatitis,due to its"simultaneous holistic and local action"and effects of tonifying the kidney yang and assisting the systemic yang,acting on the brain,heart and kidney as a whole,and meanwhile activating blood circulation,eliminating blood stasis and restoring the function of the essence chamber.This pa-per discusses the etiology and pathogenesis of chronic prostatitis with kidney deficiency and blood stasis in Chinese medicine,expounds the significance of"brain-heart-kidney-essence chamber"axis of medication,and explores the specific value and clinical application of Xiongji Formula.

AIM To investigate the consistency of chemical constituents between formula granules and standard decoction of Coptidis Rhizoma.METHODS Eighteen batches of standard decoctions were prepared,after which the extraction rate and contents,transfer rates of magnolflorine,jatrorrhizine,columbamine,epiberberine,coptisine,palmatine,berberin were determined,HPLC characteristic chromatograms were established.RESULTS There were 11 common peaks in the characteristic chromatograms of 18 batches of standard decoctions and 24 batches of formula granules with the similarities of 0.861-1.000,which were clusterd into two categories.The formula granules and standard decoction demonstrated approximated extraction rate and contents,transfer rates of index constituents.CONCLUSION The chemical constituents between formula granules and standard decoction of Coptidis Rhizoma display good consistency,which can provide references for the quality control,process research and clinical application of the former.

This study aims to screen a strain from Armillaria for the cultivation of Gastrodia elata. Specifically, Armillaria strains were isolated from different producing areas of G. elata and identified. Based on the growth characteristics of the strains and the experiment on the cultivation of G. elata, an optimal A. gallica strain was screened out. The specific process is as follows. The fungus-gro-wing materials of G. elata were collected from four producing areas and the Armillaria strains were isolated(G,Y,S,H). The strains were then identified based on morphological observation and phylogeny analysis and the commonly used strains were determined. The sucrase genotypes of the strains were identified according to our previous research findings, and the growth characteristics of the strains, such as growth rate, diameter, dry weight, and polysaccharide content of the rhizomorphs, were measured. According to the biological characteristics and sucrase genotypes, two strains were selected for the cultivation of G. elata. The tuber yield and the content of gastrodin and p-hydroxybenzyl alcohol in the tuber of G. elata were measured to select the optimal strain. The results showed that the four strains were all A. gallica. The rhizomorphs of strains G and H of the same sucrase genotype had larger/higher length, growth rate, diameter, branch number, dry weight, and polysaccharide content than those of strains S and Y of the same sucrase genotype. The tuber yield and the total content of gastrodin and p-hydroxybenzyl alcohol in tuber of G. elata cultivated with strain H were 6.528 kg·m~(-2) and 0.566%, respectively, which were 4.58 and 1.30 folds those of G. elata cultivated with strain S. Strains H and S were screened out from four strains of A. gallica based on the growth characteristics and sucrase genotype. According to the tuber yield and content of total gastrodin and p-hydroxybenzyl alcohol in the tuber of G. elata, strain H was identified as the optimal one. The findings in this study are expected to lay a basis for cultivating G. elata with high yield and quality of tubers.
Armillaria/genetics* ; Gastrodia ; Polysaccharides

OBJECTIVE: To investigate the mechanism of shikonin-induced death of human hepatocellular carcinoma SMMC-7721 cells. METHODS: Cultured SMMC-7721 cells and normal hepatocytes (L-02 cells) were treated with 4, 8, or 16 μmol/L shikonin, and the changes in cell viability was assessed using MTT assay. The levels of ATP and lactic acid in the cell cultures were detected using commercial kits. Co-immunoprecipitation and immunofluorescence staining were used to determine the relationship among pyruvate kinase M2 (PKM2), prolyl hydroxylase 3 (PHD3), and hypoxia-inducible factor-1α (HIF-1α). The expressions of PHD3, PKM2, HIF-1α, Bax, cleaved caspase-3, and Bcl-2 in SMMC-7721 cells were detected with Western blotting, and cell apoptosis was analyzed with annexin V-FITC/PI staining. The effects of RNA interference of PKM2 on PHD3 and HIF-1α expressions in SMMC-7721 cells were detected using Western blotting. RESULTS: The IC₅₀ of shikonin against SMMC-7721 and L-02 cells was 8.041 μmol/L and 31.75 μmol/L, respectively. Treatment with shikonin significantly inhibited the protein expressions of PKM2, HIF-1α and PHD3 and nuclear translocation of PKM2 and HIF-1α in SMMC-7721 cells. Coimmunoprecipitation and immunofluorescence staining confirmed that shikonin inhibited the formation of PKM2/PHD3/HIF-1α complex and significantly reduced the contents of lactic acid and ATP in SMMC-7721 cells (P < 0.05). The expressions of PHD3 and HIF-1α decreased significantly after PKM2 knockdown (P < 0.05). Shikonin treatment significantly increased the apoptosis rate, enhanced the expressions of Bax and cleaved caspase-3, and decreased Bcl-2 expression in SMMC-7721 cells (P < 0.05). CONCLUSIONS Shikonin induces apoptosis of SMMC-7721 cells possibly by inhibiting aerobic glycolysis through the PKM2/PHD3/HIF-1α signaling pathway to cause energy supply dysfunction in the cells.
Humans ; Prolyl Hydroxylases ; Carcinoma, Hepatocellular ; Caspase 3 ; bcl-2-Associated X Protein ; Liver Neoplasms ; Signal Transduction ; Apoptosis ; Adenosine Triphosphate

ObjectiveTo explore the clinical diagnosis and treatment of rare primary lumbar intervertebral space infection with Klebsiella pneumoniae and Enterobacter cloacae， and provide clinical experience for the diagnosis and treatment of this rare spinal infection. MethodsAn elderly male patient with low back pain and numbness in the left lower extremity for more than 7 months， which aggravated for more than 1 week， was diagnosed with lumbar disc herniation after laboratory and imaging examinations. After admission， the symptoms became acutely aggravated， and re-examination of lumbar enhanced MRI showed local enhancement at the posterior edge of the L3/4 intervertebral space. The VAS score was 9 points， and the lumbar JOA score was 6 points. A posterior lumbar interbody fusion of L3-L5 was performed， and L3/4 intervertebral disc specimens were collected during the operation for bacterial culture. ResultsBacterial culture results showed Klebsiella pneumoniae and Enterobacter cloacae infection. The patient was treated with sensitive antibiotics for 6 weeks after the operation， and the patient was cured during the follow-up of half a year after the operation. ConclusionFor middle-aged and elderly patients with clinical manifestations of acute severe low back pain or lower extremity pain， the possibility of spinal infection should be considered when routine laboratory and imaging examinations suggest lumbar degenerative diseases.

Oxidative stress plays a crucial role in cadmium (Cd)-induced myocardial injury. Mitsugumin 53 (MG53) and its mediated reperfusion injury salvage kinase (RISK) pathway have been demonstrated to be closely related to myocardial oxidative damage. Potentilla anserina L. polysaccharide (PAP) is a polysaccharide with antioxidant capacity, which exerts protective effect on Cd-induced damage. However, it remains unknown whether PAP can prevent and treat Cd-induced cardiomyocyte damages. The present study was desgined to explore the effect of PAP on Cd-induced damage in H9c2 cells based on MG53 and the mediated RISK pathway. For in vitro evaluation, cell viability and apoptosis rate were analyzed by CCK-8 assay and flow cytometry, respectively. Furthermore, oxidative stress was assessed by 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) staining and using superoxide dismutase (SOD), catalase (CAT), and glutathione/oxidized glutathione (GSH/GSSG) kits. The mitochondrial function was measured by JC-10 staining and ATP detection assay. Western blot was performed to detect the expression of proteins related to MG53, the RISK pathway, and apoptosis. The results indicated that Cd increased the levels of reactive oxygen species (ROS) in H9c2 cells. Cd decreased the activities of SOD and CAT and the ratio of GSH/GSSG, resulting in decreases in cell viability and increases in apoptosis. Interestingly, PAP reversed Cd-induced oxidative stress and cell apoptosis. Meanwhile, Cd reduced the expression of MG53 in H9c2 cells and inhibited the RISK pathway, which was mediated by decreasing the ratio of p-Akt^Ser473/Akt, p-GSK3β^Ser9/GSK3β and p-ERK1/2/ERK1/2. In addition, Cd impaired mitochondrial function, which involved a reduction in ATP content and mitochondrial membrane potential (MMP), and an increase in the ratio of Bax/Bcl-2, cytoplasmic cytochrome c/mitochondrial cytochrome c, and Cleaved-Caspase 3/Pro-Caspase 3. Importantly, PAP alleviated Cd-induced MG53 reduction, activated the RISK pathway, and reduced mitochondrial damage. Interestingly, knockdown of MG53 or inhibition of the RISK pathway attenuated the protective effect of PAP in Cd-induced H9c2 cells. In sum, PAP reduces Cd-induced damage in H9c2 cells, which is mediated by increasing MG53 expression and activating the RISK pathway.
Cadmium/metabolism* ; Caspase 3/metabolism* ; Potentilla/metabolism* ; Glycogen Synthase Kinase 3 beta/pharmacology* ; Proto-Oncogene Proteins c-akt/metabolism* ; Cytochromes c/metabolism* ; Glutathione Disulfide/pharmacology* ; Oxidative Stress ; Myocytes, Cardiac ; Reactive Oxygen Species/metabolism* ; Reperfusion Injury/metabolism* ; Apoptosis ; Polysaccharides/pharmacology* ; Adenosine Triphosphate/metabolism*