Mechanism of Inducing Ferroptosis in Hepatocellular Carcinoma Cells by Shugan Quyu Jiedu Prescription Based on p53/SLC7A11/GPX4 Pathway / 中国实验方剂学杂志

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.

Mechanism of Erzhiwan on Hepatocellular Carcinoma Based on Network Pharmacology / 中国实验方剂学杂志

Objective::To explore the possible mechanisms of Erzhiwan in the treatment of hepatocellular carcinoma (HCC) based on the network pharmacology. Method::The candidate active components and targets of Ligustri Lucidi Fructus and Ecliptae Herba were obtained through retrieval of the traditional Chinese medicine (TCM) systems pharmacology database (TCMSP) and literatures. Through Uniprot database and the human genome database (GeneCards), the overlapping genes of Erzhiwan and hepatocellular carcinoma were collected. The " candidate active components-targets" network of Ligustri Lucidi Fructus and Ecliptae Herba was built with Cytoscape 3.6.0 software. Drug target proteins and disease targets were mapped, and Venn map was drawn by Omicshare database. Major targets interaction network was formed by using String database and " Generate style from statistics" tool in Cytoscape 3.6.0 software. Molecular docking with active components was carried out by Systems Dock Web Site. The Gene Ontology (GO) classification enrichment analysis and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis for the targets were carried out in DAVID database. Result::Totally 21 active components, including beta-sitosterol, quercetin, luteolin, demethylwedelolactone, kaempferol, and 151 targets, including tumor necrosis factor (TNF), vascular endothelial growth factor (VEGF), N-terminal kinase (JUN), proto-oncogene (c-MYC), matrix metalloproteinase-9 (MMP-9) of Erzhiwan were collected. Erzhiwan exerts its effects on HCC mainly by acting on signal pathways, including Hepatitis B, TNF, Phosphatidylinositol-3-kinase (PI3K/Akt), tumor suppressor gene p53 and Toll-like receptor. Conclusion::Based on the methodology of network pharmacology, this study preliminarily predicted the major targets and pathways of Erzhiwan in the treatment of HCC, providing a direction for further studies.

Mechanism of action of Sini powder in treatment of liver cancer based on network pharmacology and molecular docking / 临床肝胆病杂志

ObjectiveTo investigate the mechanism of action of Sini powder in the treatment of liver cancer based on network pharmacology and molecular docking. MethodsTraditional Chinese Medicine Systems Pharmacology Database and Analysis Platform was used to obtain the compound and target of Sini powder, and the corresponding gene Symbol was obtained through Uniprot. The disease genes of liver cancer were obtained from Human Genome Database, and the genes with intersection with the target genes of Sini powder were screened out. Cytoscape3.7.1 software was used to draw the map of “traditional Chinese medicine (TCM)-compound-target” network. STRING was used to construct a protein-protein interaction (PPI) network, R studio software was used to conduct gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses on therapeutic targets, and then the results were visualized. The active component with the highest number of targets was selected as the ligand, and the target with the highest degree in the PPI network was selected as the receptor, so as to predict the structure of receptor-ligand complex and the amino acid residues that bind to each other. ResultsIn this study, 91 core targets and 141 relevant active components of Sini powder were screened out, among which quercetin and kaempferol were the main active components in the treatment of liver cancer. TP53 and HSP90AA1 were the main therapeutic targets. The GO enrichment analysis obtained 1007 items which met the screening criteria, which were mainly involved in the biological processes of antioxidation reaction, activity regulation of protein serine and threonine kinase, and cellular stress response. The KEGG enrichment analysis obtained 102 pathways, which mainly regulated the hepatitis B pathway and the PI3K-Akt signaling pathway in the prevention and treatment of liver cancer. The results of molecular docking showed a synergistic antitumor effect between the crystal structure domains VAL147, CYS220, GLU221, and PRO222 of quercetin-TP53. ConclusionThis study reveals the mechanism of action of Sini powder in the treatment of liver cancer by acting on multiple targets and signaling pathways, which provides a theoretical basis for biological experiments.

HEAD-US-C quantitative ultrasound assessment scale in evaluation of joint damage in patients with moderate or severe hemophilia A received on-demand versus prophylaxis replacement therapy / 中华血液学杂志

Objective: To explore the evaluation of joint injury by HEAD-US-C (Hemophilic Early Arthropathy Detection with UltraSound in China, HEAD-US-C) in patients with moderate or severe hemophilia A treated with prophylaxis vs on-demand. Methods: The patients from June 2015 to July 2017 with moderate or severe hemophilia A were examined by ultrasound imaging of the elbows, knees and ankles; Meanwhile the HEAD-US-C ultrasound assessment scale and hemophilia joint health score scale 2.1 (HJHS2.1) were used to score the joint status. The correlation between the HEAD-US-C and HJHS score was performed in prophylaxis group and on-demand group patients, respectively. Results: A total of 925 cases of joint ultrasonography were conducted in 70 patients with moderate or severe hemophilia A. Among patients with moderate hemophilia, the median (IQR) of HEAD-US-C score and HJHS score in on-demand group were significantly higher than those in the prophylaxis group[1 (0, 6) vs 0.5 (0, 3) , z=0.177, P=0.046],[2 (0, 4) vs 2 (0, 3) z=0.375, P=0.007], even though there was no significant difference of the median (IQR) number of annualized target joints bleeding episodes between on-demand and prophylaxis groups[1 (0, 7) vs 1 (0, 5) , z=1.271, P=0.137]. Unlike in moderate cases, on-demand treatment group had more annualized target joints bleeding episodes than prophylaxis group among patients with severe hemophilia[3 (0, 8) vs 2 (0, 8) , z=0.780 P=0.037]. The prophylaxis group compared favorably with on-demand therapy group in terms of HEAD-US-C score[1 (0, 6) vs 4 (0, 7) , z=2.189, P=0.008], and HJHS score[2 (0, 5) , 4 (1, 6) , z=3646, P<0.001]for the severe hemophilia patients. The positive correlation between HEAD-US-C score and HJHS score was identified (P<0.05) , whether on-demand treatment or prophylaxis groups. The correlation coefficient between HEAD-US-C score and HJHS score in on-demand treatment and prophylaxis groups were 0.739 (95% CI 0.708-0.708) , 0.865 (95% CI 0.848-0.848) respectively, and 95% CI didn't overlap (P<0.05) , indicating that the correlation coefficient in prophylaxis group had stronger correlation than that in on-demand group. Conclusions: Clinical effects of prophylaxis were significantly better than those of on-demand treatment in patients with moderate or se-vere haemophilia A. HEAD-US-C scoring system could effectively evaluate joints damage in hemophilia A patients treated with on-demand or prophylaxis, companied by significantly positive correlation with HJHS clinical evaluation system, and provided objective index for clinical effect assessment.

Impact of anchorage implant design factors on its primary stability / 中华口腔医学杂志

During the past decade, the anchorage implants have been widely used in the orthodontic treatment. Many scholars have studied the influence of different factors on anchorage implant's primary stability, including anchorage implant's material properties, structural design, surgical procedure, bone condition, loading force's magnitude and direction. This article is to review the influence of anchorage implant's shape, dimension, neck design and thread design on its primary stability.

Loss expression of active fragile sites genes associated with the severity of breast epithelial abnormalities / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>WWOX and FHIT are two candidate tumor suppressor genes located in active fragile sites, the damage of which has been associated with the development of breast cancer. The association of the expression of these genes and the development of breast cancer has not been fully explored. We evaluated mRNA and protein expression of WWOX and FHIT in breast tissue with normal histological appearances, atypical ductal hyperplasia, ductal carcinoma in situ, and invasive cancer to see if a progressive decline in expression was present.</p><p><b>METHODS</b>Reverse transcription-polymerase chain reaction and Western blotting were used to evaluate the specimens for mRNA and protein expression, including 28 specimens with normal tissue, 28 specimens with atypical ductal hyperplasia, 33 specimens with ductal carcinoma in situ, and 51 specimens with invasive ductal carcinoma.</p><p><b>RESULTS</b>Compared with in situ and invasive cancer specimens, both normal and atypical hyperplasia specimens had greater rates of detectable mRNA (WWOX rate ratio = 2.95, 95% CI 1.24 - 7.08; FHIT rate ratio = 4.58, 95% CI 1.82 - 11.81) and Western blotting detectable protein (WWOX rate ratio = 4.12, 95% CI 1.63 - 10.73; FHIT rate ratio = 3.76, 95% CI 1.44 - 10.06). For both proteins, differences between normal and atypical hyperplasia specimens and between in situ and invasive carcinoma specimens were explainable by chance (P > 0.05 for each analysis). Within each histological category, differences among fractions of specimens showed that FHIT and WWOX mRNA and protein expression were explainable by chance (P > 0.05 for each analysis).</p><p><b>CONCLUSION</b>Expression of FHIT and WWOX decreases along with breast tissue progress from a normal histological appearance to atypical ductal hyperplasia, in situ cancer, and the final invasive cancer.</p>

Clinical evaluation of laparoscopic common bile duct exploration in 587 cases / 中华外科杂志

<p><b>OBJECTIVE</b>To summarize the experience of laparoscopic common bile duct exploration.</p><p><b>METHODS</b>The clinical data of 587 cases who underwent laparoscopic common bile duct exploration from June 1992 to May 2006 were analyzed.</p><p><b>RESULTS</b>The surgery was successful in 585 cases (99.7%), 2 cases were converted to open common bile duct exploration. The duration of operation was 60 approximately 230 min (averaged 85 min), the complications consisted of biliary fistula (n=13), injury of the duodenum (n=1), abscess of drainage tube orifice (n=1), titanium clip discharging out from T tube (n=3), residual common bile duct stones (n=35). The patients could take food and walk on the second postoperative day and average postoperative hospital stay was 4.6 days.</p><p><b>CONCLUSIONS</b>Laparoscopic common bile duct exploration is a safe and effective procedure in treating the calculus of bile duct.</p>

Expression and clinical significance of certain apoptosis and angiogenesis factors in hepatocellular carcinoma / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the expression and clinical significance of some apoptosis and angiogenesis factors in hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>The expression of p53, Survivin, matrix metalloproteinases (MMP)-2, MMP-9 and vascular endothelial growth factor (VEGF) in 90 specimens of HCC was detected by immunohistochemistry. Correlations of the factors to the recurrence of HCC after hepatectomy were analyzed.</p><p><b>RESULTS</b>The positive rate of p53, Survivin, MMP-2, MMP-9 and VEGF in HCC tissue was 33.3%, 51.1%, 60.0%, 37.8% and 76.7%, respectively. Of the 5 factors, positive correlation only occurred between the expression of MMP-2 and VEGF, MMP-9 and VEGF. The expression of MMP-2, MMP-9 and VEGF was correlated to the recurrence of HCC. The 1-, 2-, and 3-year tumor-free survival rates were significantly higher in MMP-2 (-) group than in MMP-2 (+) group, and the same results were found with MMP-9 and VEGF. Multivariate analysis revealed that macroscopically disseminated nodules, tumor micrometastasis, serum alpha fetal protein (AFP) level, the expression of MMP-9 and VEGF were independent recurrence risk factors in HCC.</p><p><b>CONCLUSIONS</b>Neither p53 nor Survivin is correlated to the recurrence of HCC; MMPs and VEGF are correlated to the recurrence, and can be used to estimate the risk of postoperative recurrence of HCC.</p>

Expression and clinical impact of vascular endothelial growth factor and matrix metalloproteinase-2 in hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the expression and clinical impact of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 (MMP-2) in hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>Ninety specimens obtained from HCC patients were examined immunohistochemically using anti-VEGF and anti-MMP-2 monoclonal antibodies.</p><p><b>RESULTS</b>The positive rates of VEGF and MMP-2 expression in HCC tissues were 76.7% and 60%, respectively. The expression of MMP-2 in HCC tissues was positively correlated with the expression of VEGF (r(s) = 0.32) and both were positively correlated with recurrence (or metastasis) after hepatectomy (r(s) = 0.31, r(s) = 0.32). 2-year tumor-free survival rates of VEGF- group, VEGF+ group and VEGF++ group were 71.4%, 43.5%, 30.4%, respectively, (P < 0.01), while MMP-2- group 66.7% and MMP-2+ group 32.8% (P < 0.01). Multivariate analysis revealed that the expression of VEGF and MMP-2 in HCC tissues, tumor microthrombus and pre-operative dissemination to lymph nodes were independent recurrence (or metastasis) risk factors.</p><p><b>CONCLUSION</b>The expression of VEGF and MMP-2 in HCC tissues, and clinicopathological features (tumor microthrombus and pre-operative dissemination to lymph nodes), could be regarded as valuable indicators for prediction of recurrence (or metastasis) risk in HCC patients.</p>

Observation on change of lipidemia and lipoprotein in wistar rats wi th fluorosis / 中国地方病学杂志

Objective　To observe the change of lipidemia and lipoprotein in wistar rats with fluorosis.Methods　14 wistar rats were fed normal food with hyper concentration of NaF water (100 mg/L) for six months,the TC,TG,HDL-c,LDL-c,APO AI APO B and AI,R-CHE were measured in serum.Results　The result show that there are an increace of T C,TG,HDL-c,LDL-c,APO AI，APO B and AI,R-CHD compared with normal rats (P <0.05).Conclusions　The result indicate that the fluorosis can increase the lipidemia and lipoprotein in rats with fluorosis and lead to athero selerosis.

The diagnosis and treatment of hepatic focal nodular hyperplasia / 中国普通外科杂志

Objective To explore the clinical diagnosis and management of focal nodular hyperplasia (FNH) of the liver. Methods Forty-two FNH cases treated in the past 9 years were studied retrospectively. The clinical and pathologic data were reviewed. Results Preoperative liver function test and AFP were normal. The preoperative radiography in FNH was usually not specific, with less than 50% cases were suggestive of FNH of the liver. Surgical resection resulted in a permanent cure with no significant postoperative complications. More than one year follow-up found recurrence in one case. Conclusion Clinical, laboratory and radiological findings when combined could help in establishing tentative diagnosis of FNH. Surgery is recommended in cases with equivocal diagnosis or in fear of hepatocellular carcinoma.