Preliminary study of PPARA regulate the ferroptosis of hepatocellular carcinoma to prevent disease deterioration / 中华预防医学杂志

To explore whether PPARA is involved in the process of ferroptosis in hepatoma cells, peroxisome proliferator activated receptor (PPARA) was comprehensively analyzed in hepatocellular carcinoma (HCC) through public database and experimental data, including the expression, the functions and the potential roles of tumor progression. The research design is experimental research,data analysis based on bioinformatics and cell experiment. From January 2022 to August 2022, relevant cell experiments were conducted in the Basic Medical Laboratory of the General Hospital of the Southern Theatre of the Chinese People's Liberation Army. The expression and the correlation with clinicopathologic features of PPARA in HCC were analyzed by The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. To study the protein expression of PPARA in HCC and normal tissues through the Human Protein Atlas (HPA). The protein-protein interaction (PPI) network between PPARA and the core factor of ferroptosis was constructed based on Search Tool for the Retrival of Interacting Genes/Protein (STRING) database, then, the correlation between PPARA and the core gene Glutamate-cysteine Ligase Catalytic Subunit (GCLC) was analyzed by Gene Expression Profiling Interactive Analysis (GEPIA). Assessed the expression of PPARA in HCC cell lines SK-HEP-1, SMMC-7721, MHCC-97H, BEL-7402 and normal liver cell L02 by Western Blot (WB) and the changes of PPARA expression after 48h treatment with ferroptosis inducer Erastin were observed. Single factor analysis of variance was used to compare the expression of PPARA between groups in GEPIA database. The expression of PPARA in GSE25097 and GSE112790 data was compared by rank sum test. Survival analysis was performed using time series test method. The difference of PPARA expression between clinical and pathological features was compared using the Kruskal-Wallis test. The correlation between the expression of GCLC and PPARA was compared by the method of Spearman correlation. The expression of PPARA in cell lines was compared by paired T test. The results showed that the RNA and protein expression of PPARA in HCC was lower than that in normal tissues (P<0.05). PPARA alterations were correlated with patient clinicopathological features and prognosis (P<0.05). The PPI constructed by STRING database suggests that PPARA interact with the key factors of ferroptosis, such as NFE2 like bZIP transcription factor 2 (NFE2L2), Heme Oxygenase 1 (HMOX1), Tumor Protein P53 (TP53), GCLC, Dipeptidyl Peptidase 4 (DPP4), Citrate Synthase (CS), Arachidonate 15-Lipoxygenase (ALOX15) and Acyl-CoA Synthetase Long Chain Family Member 4 (ACSL4). Furthermore, the PPARA was significantly associated with GCLC validated via GEPIA database(R=0.6, P<0.05). The expression of PPARA increased after treatment with ferroptosis inducer Erastin for 48 h by WB. In conclusion, the expression of PPARA is lower in HCC with a poor prognosis. PPARA interacts with GCLC in regulating ferroptosis in HCC.

Antiproliferation activity of a naphthoquinone derivative against vascular smooth muscle cells / 中国药理学通报

Aim: To explore the effect of a synthetic naphthoquinone derivative on the proliferation of rat aortic vascular smooth muscle cells(RAVSMCs) stimulated by platelet-derived growth factor BB (PDGF-BB) and to clarify its mechanism underlying the anti-proliferative effect. Methods: The influence of the synthetic naphthoquinone derivative on cell cycle progression and main signal transduction pathway of cell cycle induced by PDGF-BB were investigated by cell proliferation assay, [3H] thymidine incorporation test, cell cycle process analysis and immunoblotting assay. Results: S phase cell percentage in the cell cycle was reduced, while G0/G1 phase cell percentage was increased by the synthetic naphthoquinone in a dose-dependent (0. 1, 0. 5, 1 μmol · L-1) manner. Moreover, DNA synthesis also decreased. The inhibitory rate of PDGF-BB-induced RAVSMCs proliferation achieved the maximum of 44. 4% after 24 h pre-treatment by the synthetic naphthoquinone derivative at the concentration of 1 μmol · L-1. The phosphorylation of extracellular regulated kinase l/2(ERKl/2), Akt, phospholipase C (PLC) γ1 and PDGF receptor β(PDGFRβ) induced by PDGF-BB was significantly decreased (P < 0. 01) by addtion of 1 μmol · L-1 synthetic naphthoquinone derivative. Conclusions: The synthetic naphthoquinone derivative exhibits anti-proliferation activity against RAVSMCs induced by PDGF-BB via blocking the transformation of G0/G1 phase cell into S-phase cell in the cell cycle. The mechanisms might be related to its down-regulatory effect on phosporalation level of ERK1/2, Akt, PLCγ1 and PDGFRβ.

Influence of alisol B 23-acetate on blood glucose in type 2 diabetic mice / 中国药理学通报

Aim To explore whether alisol B 23-acetate possesses the therapeutic potential for treatment of type 2 diabetes mellitus ( T2DM ). Methods T2DM mouse model was established by combined administration of streptozotocin and nicotinamide. After three weeks of oral administration of rosiglitazone or alisol B 23-acetate, the blood glucose of type 2 diabetic mice was measured. Oral glucose tolerance test(OGTT) was carried out the next day. Rosiglitazone was chosen as positive drug. 2-[N-(7-nitrobenz-2-oxa-l, 3-diazol-4-yl) amino]-2-deoxy-D-glucose (2-NBDG) uptake assay in adipocytes was adopted to test whether alisol B 23-acetate had effect on glucose uptake in cells. 3T3-Ll pre-adipocytes differentiation model was performed to evaluate whether alisol B 23-acetate promoted adipo-genesis. Results Mice exhibited significantly higher blood glucose concentration after intraperitoneal injection of streptozotocin and nicotinamide for three weeks, as examined by blood glucose concentration on day 21 and OGTT on day 22, compared with normal mice in blank control group. After orally administrating alisol B 23-acetate at dose of 5 mg • kg-1 , 10 mg • kg-1 ,20 mg • kg-1 daily for three weeks, respectively, or orally administered rosiglitazone at dose of 10 mg • kg-1 daily for three weeks, blood glucose greatly decreased in type 2 diabetic mice. Moreover, insulin resistance was also improved to a certain degree during OGTT. Furthermore, alisol B 23-acetate not only increased insulin-induced glucose uptake in adipocytes at the concentration of 30 mmol • L-1 glucose, but also accelerated 3T3-L1 pre-adipocytes differentiation process at concentration of 1 μmol • L-1 and 10 μmol • L-1 . Conclusions Alisol B 23-acetate reduces blood glucose of type 2 diabetic mice, promotes pre-adipocyte differentiation and increases glucose uptake in adipocytes; however, the mechanism of action needs further exploration.

Hypoglycemic effect of alismoxide in type 2 diabetic mice / 中国药理学通报

Aim To investigate the hypoglycemic effect of alismoxide. Methods Type 2 diabetes mellitus (DM) mouse model induced by combined administration of streptozotocin and nicotinamide was adopted. Three weeks later, blood glucose of blank control group and type 2 diabetic mouse model group was measured on day 21 , and oral glucose tolerance test(OGTT) was carried out on day 2 2 , respectively. After type 2 diabetic mouse model was successfully established, rosiglitazorie was chosen as positive drug. Oral administration of rosiglitazone at dose of 10 mgk g-1 daily was performed for three weeks in positive group. Oral administration of alismoxide at dose of 5 , 10 and 20 mg kg"1 daily for three weeks was carried out in alismoxide different dose group, respectively. Furthermore, influence of alismoxide on differentiation was investigated in 3T3-L1 pre-adipocytes, and Oil red 0 staining was adopted. Results Not only blood glucose was decreased by alismoxide in type 2 DM mice, but also hypoglycemic trend was exhibited during OGTT. Furthermore, at concentration of 0. 5 and 1 fimol L " 1 , alismoxide promoted 3T3-L1 pre-adipocyte differentiation. Conclusions It suggests that alismoxide might possess hypoglycemic property and accelerate pre-adipocyte differentiation; however, the mechanism involved needs further study.

Synthesis and anti-tumor activity of ginsenoside Rh_2 caprylic acid monoester / 中国中药杂志

Ginsenoside Rh_2,firstly isolated from red ginseng,is protopanaxadiol type of steroidal saponin. Rh_2 possessed variety of activities,but bioavailability of oral administration Rh_2 was extremely low due to poor absorption. Moreover,ginsenoside Rh_2 exhibited toxicity on human normal cells. Therefore,to improve stronger anti-tumor activity and attenuate toxicity,it was essential to design and optimize chemical structure of ginsenoside Rh_2. Through n-octanoylchloride modifications,a novel ester derivative of ginsenoside Rh_2 named caprylic acid monoester of Rh_2( C-Rh_2) was designed and synthesized. Structure of novel ginsenoside derivative was identified by1 D and 2 D NMR,as well as ESI-MS analyses. Anti-tumor effect of C-Rh_2 was tested on H22 tumor bearing mice. C-Rh_2 displayed certain anti-tumor activities and exhibited less toxicity than Rh_2. In the present study,C-Rh_2 as ester form of ginsenoside Rh_2 showed better anti-tumor activity and less toxicity,but the specific mechanism needs further investigation.

Epidemiological survey of high-risk human papillomavirus among 2501 woman / 南方医科大学学报

<p><b>OBJECTIVE</b>To survey the prevalence of high-risk human papillomavirus (HPV) in woman in Guangzhou during the period from 2013 to 2014.</p><p><b>METHODS</b>A total of 2501 women in Guangzhou seeking medical attention in our hospital underwent high-risk HPV genotype screening of cervical specimens using real-time PCR.</p><p><b>RESULTS</b>The prevalence of high-risk HPV infection among the women was 14.85% (146/983) in the year 2013, similar to the rate of 14.56% (221/1518) in 2014 (Χ(2)=0.041, P=0.839); no significant differences were found in the high-risk HPV infection rates between different age groups in either 2013 (Χ(2)=2.916, P=0.572) or 2014 (Χ(2)=6.494, P=0.165). The constituent ratio of the 13 types of high-risk HPV showed no significant difference between 2013 and 2014 (Χ(2)=11.872, P=0.452). The 13 HPV genotypes detected, listed in a descending order of the constituent ratios, included HPV-52, -16, -58, -56, -39, -51, -68, -59, -31, -35, -18, -33 and -45 in 2013, and were HPV-52, -16, -58, -68, -18, -51, -56, -39, -31, -33, -59, -35 and-45 in 2014.</p><p><b>CONCLUSION</b>We report a high prevalence of high-risk HPV among women in Guangzhou, which suggests the necessity of screening for high-risk HPV-DNA among women at all ages for prevention and early detection of cervical cancer.</p>

Comparative analysis of ginsenosides from red ginseng / 中国中药杂志

<p><b>OBJECTIVE</b>To determine ginsenoside Rg1, Re, Rb1, Rc, Rb2, Rd from main root and root hair of red ginseng of different specifications.</p><p><b>METHOD</b>Ultrasonical extraction and reversed phase high performance liquid chromatography were applied.</p><p><b>RESULT</b>The total contents of six ginsenosides from main root 15 roots (percent 500 g), 20 roots, 30 roots and root hair are 1.21%, 1.46%, 1.54% and 8.16%, respectively.</p><p><b>CONCLUSION</b>The results showed that the bigger the volume of ginseng root, the less the content of ginsenoside.</p>

Depressant Effect of siRNA on the Expression and Replication of Hepatitis B Virus in HepG2.2.15 Cell / 实用儿科临床杂志

Objective To explore the siRNA as a new antiviral therapy,evaluate the inhibition effect of siRNA based on vector on the HBV of HepG2.2.15 cell,and observe the side effect and toxicity of siRNA vector on cells and the off-target effect of siRNA.Methods Three pairs of siRNA duplexes targeting HBV C gene were designed as double strands,and the duplex were annealed and ligated into the p-Silencer-Cmv 4.1-hygro vector.The ligation products were used to transform JM109 cells.The clones with shRNA were obtained,and the vectors were purified.After the initial identification of the vector with agarose gel and the size of the inserted sequence got examined by native polyacrylamide gel electrophoresis,furthermore the sequencing was further carried out.The recombinant plasmids were purified with ultrapure Midipreps DNA Purification System.Then HepG2.2.15 cells were transfected with the plasmid mixed with siPort XP-1.The expression of HBsAg and HBeAg were detected by immunofluorescence and Western blot,and the HBV RNA was investigated by RT-PCR.Furthermore the real-time quantitive PCR was carried out to detect the changes of HBV DNA.In order to evaluate the toxicity of the shRNA,MTT was used to examine the growth rate and curve of cells.The ELISA was performed to detect the changes of interferon-? (IFN-?).Results The Western blot showed that the HBsAg and HBeAg protein were suppressed with (81.15?0.69)%,(88.12?0.92)% respectively by vector p-C2 on the third day of post-transfection.It had the similar result indicated by immunofluorescence.And the RT-PCR showed that the specific siRNA targeting HBV C gene could markedly suppress the expression of HBV mRNA and the HBV C gene mRNA was inhibited with 96.9%.The real-time quantitive PCR showed that the specific functional siRNA could markedly suppress HBV DNA copy with two orders of magnitude,while the siRNA vector had no effect on the growth of cell showed by MTT detection.Compared with the non-transfected group and p-NC group,the IFN-? level was almost the same with siRNA p-C1,p-C2,p-C3 groups.Conclusions The siRNA based on the expression vector can suppress the expression and replication of HBV in HepG2.2.15 cell.The inhibition effect was specific and had a certain dependency on siRNA concentration.No toxicity effect was found in the study.And the drug resistance wouldn′t happen because the silence was based on the split of gene.

The effects of banxia houpu decoction on a chronic mild stress model of depression / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of Banxia Houpu Decoction on a chronic mild stress model of depression and investigate the antidepressive mechanism.</p><p><b>METHOD</b>With consumption of a 1% sucrose solution as an index, and by subjecting rats to a variety of mild stressors for a prolonged period of time, a chronic mild stress model was developed. The levels of the blood lipid were measured by blood lipid kits, the natural kill (NK) cell activity in the spleen was measured with the method of the enzyme lactate dehydrogenase (LDH) released, the superoxide dismutase (SOD) activity in red blood cell was assayed by the Autoxidation of Pyrogallol method, the NO synthase (NOS) activity in serum and tissue was measured by NOS kits, and the content of malondialdehyde(MDA) was measured by MDA kits.</p><p><b>RESULT</b>Banxia Houpu Decoction significantly increased the consumption of sucrose solution, increased the level of the high density lipoprotein (HDL-C), decreased the level of the Triglyceride(TG) in serum, enhanced the activity of the NK in spleen, decreased the activity of the SOD in red blood and the activity of the NOS in serum and tissue, and reduced the content of MDA in tissue by effect on lipid Peroxidation in CNS model of depression.</p><p><b>CONCLUSION</b>Banxia Houpu Decoction has antidepressant effect in different ways.</p>