[Abstract] Objective: To investigate the effect of polyadenosine diphosphate ribose polymerase 1 (PARP1) on the proliferation and 5-FU resistance of gastric cancer cells and its potential mechanism. Methods: The tumor tissues and corresponding paracancerous tissues of 72 patients with gastric cancer who were treated in Central Hospital of Enshi Tujia and Miao Autonomous Prefecture from May 2018 to December 2019 were collected. AGS cells were transfected with siFUT8 to knock down FUT8 gene expression. qPCR and immunohistochemistry were used to detect the expression of PARP1 in gastric cancer and adjacent tissues. CCK-8, flow cytometry, and colony formation assay were employed to detect the effects of AG14361 on the proliferation, apoptosis and colony formation of AGS cells. MTT assay was used to detect the effect of AG14361 on the 5-FU sensitivity of gastric cancer cells. The overall distribution of differential genes in AGS cells treated with AG14361 was analyzed by mRNA sequencing, and related signaling pathways were analyzed by KEGG enrichment. qPCR and WB were used to detect the expression of α-1,6-fucosyltransferase (FUT8) in AGS cells and the interference effect of FUT8. CCK-8, flow cytometry, and colony formation assay were employed to detect the effects of AG14361 on the proliferation, apoptosis, and colony formation of AGS cells disturbed by siFUT8. Results: Compared with paracancer tissues, PARP1 expression was significantly increased in gastric cancer tissues (P<0.001). AG14361 can significantly inhibit the proliferation and colony formation of AGS cells, thus promoting the apoptosis of AGS cells (all P<0.01). AG14361 treatment reduced the IC50 of 5-FU against gastric cancer cells, especially against AGS cells, with IC50 decreased by more than 60%. mRNA sequencing results showed that FUT8 was a key glycosyltransferase of AG14361 in inhibiting the proliferation of AGS cells (P<0.05). Compared with the siNC group, treatment of AG14361 with IC50 significantly reversed the promotion of AGS cells proliferation caused by inerference with FUT8, promoted apoptosis and BAX protein expression, decreased Bcl2 protein expression and inhibited the increase in AGS cell colony formation caused by interference with FUT8 (all P<0.01). Conclusion: PARP1 can promote malignant transformation of gastric cancer cells by regulating N-glycosyltransferase FUT8. AG14361 can enhance the chemotherapy sensitivity of 5-FU, and PARP1 may become a potential target for gastric cancer treatment.

Objective: To investigate the effects of sulforaphane on lipopolysaccharide (LPS)- induced cardiac myocytes hypertrophy of rats and study its possible mechanism. Methods: The hypertrophic primary cardiac cells of neuonatal rats were divided into control group, LPS-induced group, JAK2 inhibitor group, low-, middle-, and high-dose sulforaphane groups. The cells in all groups, except for control group, were intervened by LPS (1 mg/L). The low-, middle-, and high-dose sulforaphane groups were treated with 10, 20, 40 μg/ml sulforaphane respectively. The JAK2 inhibitor group was treated with 10 nmol/L of JAK2 inhibitor for 24 h. Computer photograph analysis system was used to determine the cardiomyocyte volume, BCA kit was used to analyze the total protein concentration, ELISA kit was used to observe the expression levels of inflammatory cytokine (including IL-6 and TNF-α), and the Western Blot technology was used to analyze the expression levels of the related proteins in JAK2/STAT3 signaling pathway. Results: Compared with control group, the cell volume (1 635.71 ± 154.84 μm³, 1 450.06 ± 140.13 μm³, 1 194.51 ± 134.76 μm³ vs. 1 854.28 ± 181.37 μm³) and the levels of protein (20.14 ± 2.11 μg, 17.59 ± 1.64 μg, 14.27 ± 1.47 μg vs. 23.17 ± 2.56 μg) in low-, medium-, high- dose of sulforaphane groups were significantly decreased (P<0.05). The levels of IL-6 (1 410.08 ± 255.17 pg/ml, 1 065.61 ± 210.37 pg/ml, 790.09 ± 140.28 pg/ml vs. 1 804.07 ± 275.34 pg/ml) and TNF-α (164.16 ± 27.51 pg/ml, 130.13± 22.08 pg/ml, 92.27 ±12.16 pg/ml vs. 182.42 ± 30.37 pg/ml) in low-, medium-, high- dose of sulforaphane groups were significantly decreased (P<0.05). The expression levels of p-JAK2/JAK2 (0.39 ± 0.05, 0.27 ± 0.04, 0.21 ± 0.03 vs. 0.54 ± 0.07) and p-STAT3 (0.47 ± 0.05, 0.25 ± 0.03, 0.18 ± 0.03 vs. 0.53 ± 0.06) in low-, medium-, high- dose of sulforaphane groups were also significantly decreased (P<0.05). Conclusions Sulforaphane has a protective effect on LPS-induced cardiac hypertrophy, which is partially via inhibiting inflammatory response via suppressing the activation of JAK2/STAT3 signaling pathway.

Multifocal electroretinogram(mf-ERG)is a common type of electroretinogram. It can objectively, accurately and quantitatively detect the function of each tiny part of the retina, so it is widely used in clinical, especially in retinal diseases. It not only plays an important role in the diagnosis and efficacy evaluation of retinal diseases, but also has an important value in the follow-up and prevention of diseases, even the scope of its application can be extended to basic research. Therefore, this paper reviews the application of multifocal electroretinogram in retinal diseases.

OBJECTIVES: To study the effect of ligustrazine injection on mitophagy in neonatal rats with hypoxic-ischemic encephalopathy (HIE) and its molecular mechanism. METHODS: Neonatal Sprague-Dawley rats, aged 7 days, were randomly divided into a sham-operation group with 8 rats, a model group with 12 rats, and a ligustrazine group with 12 rats. The rats in the model group and the ligustrazine group were used to establish a neonatal rat model of HIE by ligation of the left common carotid artery followed by hypoxia treatment, and blood vessels were exposed without any other treatment for the rats in the sham-operation group. The rats in the ligustrazine group were intraperitoneally injected with ligustrazine (20 mg/kg) daily after hypoxia-ischemia, and those in the sham-operation group and the model group were intraperitoneally injected with an equal volume of normal saline daily. Samples were collected after 7 days of treatment. Hematoxylin and eosin staining and Nissl staining were used to observe the pathological changes of neurons in brain tissue; immunohistochemical staining was used to observe the positive expression of PINK1 and Parkin in the hippocampus and cortex; TUNEL staining was used to measure neuronal apoptosis; Western blotting was used to measure the expression levels of the mitophagy pathway proteins PINK1 and Parkin and the autophagy-related proteins Beclin-1, microtubule-associated protein 1 light chain 3 (LC3), and ubiquitin-binding protein (P62). RESULTS: Compared with the sham-operation group, the model group had a significant reduction in the number of neurons, an increase in intercellular space, loose arrangement, lipid vacuolization, and a reduction in Nissl bodies. The increased positive expression of PINK1 and Parkin, apoptosis rate of neurons, and protein expression levels of PINK1, Parkin, Beclin1 and LC3 (P<0.05) and the decreased protein expression level of P62 in the hippocampus were also observed in the model group (P<0.05). Compared with the model group, the ligustrazine group had a significant increase in the number of neurons with ordered arrangement and an increase in Nissl bodies, significant reductions in the positive expression of PINK1 and Parkin, the apoptosis rate of neurons, and the protein expression levels of PINK1, Parkin, Beclin1, and LC3 (P<0.05), and a significant increase in the protein expression level of P62 (P<0.05). CONCLUSIONS Ligustrazine can alleviate hypoxic-ischemic brain damage and inhibit neuronal apoptosis in neonatal rats to a certain extent, possibly by inhibiting PINK1/Parkin-mediated autophagy.
Rats ; Animals ; Hypoxia-Ischemia, Brain/metabolism* ; Animals, Newborn ; Rats, Sprague-Dawley ; Beclin-1 ; Autophagy ; Ubiquitin-Protein Ligases/metabolism* ; Protein Kinases/metabolism*

OBJECTIVE: To investigate the protective effects of Ginkgo biloba extract(GBE) on paracetamol(APAP)-induced acute hepatic injury in mice and its mechanism. METHODS: Thirty mice were randomly divided into control group, model group, GBE low, medium and high-dose(50,100,and 200 mg·kg)groups,with 6 mice in each group. All mice except control group were administered with APAP(300 mg/kg)for one time by intraperitoneal injection. The mice in GBE low, medium and high-dose groups were intragastric administered with GBE for 2 d consecutively, then samples were harvested for analysis. The appearance and pathology of liver were observed. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in serum and the levels of superoxide dismutase (SOD), myeloperoxidase(MPO), glutathione (GSH) and malondialdehyde (MDA) in hepatic tissue were measured. Western blot was used to detect the protein expressions of Nrf2 and HO-1. RESULTS: Compared with control group, in model group, the appearance and pathology of liver were bad, the levels of ALT,AST,TNF-α and IL-6 in serum were increased significantly(<0.01),the levels of GSH and SOD were decreased while the levels of MDA and MPO were increased in hepatic tissue(<0.01), the expressions of Nrf2 and HO-1 were increased in hepatic tissue(<0.05). Compared with model group, in GBE groups, the appearance and pathology of liver were improved, the levels of ALT,AST,TNF-α and IL-6 in serum were decreased significantly(<0.01), the levels of GSH and SOD were increased while the levels of MDA and MPO were decreased in hepatic tissue(<0.01), the expression of Nrf2 and HO-1 were increased in hepatic tissue(<0.05). The high-dose of GBE possessed the most obvious treatment effect among them. CONCLUSIONS GBE may play a protective role in APAP-induced acute hepatic injury through Nrf2/HO-1 pathway.
Acetaminophen ; Alanine Transaminase ; Animals ; Aspartate Aminotransferases ; Chemical and Drug Induced Liver Injury ; Ginkgo biloba ; Liver ; Malondialdehyde ; Mice ; Oxidative Stress ; Plant Extracts

Polysaccharide of Balanophora involucrata Hook. f. (BPS), the major component of Balanophora involucrata Hook. f., was confirmed the protective effect on liver injury in our previous study. This research aimed to investigate the protective mechanism of BPS on experimental liver injury by attenuating cell ferroptosis through modulating solute carrier family 7 member 11/glutathione peroxidase 4 (SLC7A11/GPX4) pathway. The animal experiment was approved by the Experimental Animal Ethical Committee of Hubei Minzu University and all rats had received human care in compliance with the institutional animal care guidelines. Rats were given intraperitoneal injection of (D-galactosamine, D-GalN) solution (800 mg·kg^-1) one time to establish the acute liver injury model. The results showed aspartate amino transferase (AST), alanine aminotransferase (ALT) and 4-hydroxynonenal (4-HNE) levels in serum were decreased, and the contents of reactive oxygen species (ROS), Fe²⁺, malondialdehyde (MDA) and lipid peroxide (LPO) in liver tissues also decreased and glutathione (GSH) level increased after BPS administration with 200 mg·kg^-1. Besides, BPS reduced iron deposition and increased the expression of SLC7A11 and GPX4 proteins in liver tissue. In conclusion, BPS ameliorated experimental liver injury by alleviating cell ferroptosis through SLC7A11/GPX4 pathway. The present study pointed to the possibility of utilizing BPS for protection against liver injury in clinic.

Objective:To explore the effect of trillin on oxidative stress response and nuclear factor E2-related factor 2/antioxidant response element (Nrf2/ARE) pathway in rats after spinal cord injury (SCI). Methods:A total of 108 male Sprague-Dawley rats were randomly divided into sham group (n = 36), model group (n = 36) and trillin group (n = 36), each group was divided into one day, three days and seven days subgroups, with twelve rats in each subgroup. The SCI model was established by modified Allen's heavy strike method in the model group and the trillin group, but no obvious injury in the sham group. The trillin group was given trillin 200 mg/kg every day, and the same amount of normal saline was given in the sham group and model group, twice a day. BBB score was performed one day, three days and seven days after modeling. Morphological changes were tested by Nissl's staining, and the changes of malonaldehyde (MDA) content and superoxide dismutase (SOD) activity were detected by ELISA seven days after modeling. The expression of Nrf2, Kelch like ECH associated protein 1 (Keap1), NAD(P)H quinone oxidoreductase (NQO1) and haemoxygenase 1 (HO-1) were detected by Western blotting one day, three days and seven days after modeling. Results:Compared with the model group, BBB scores increased (P < 0.05); the structure of spinal cord was more complete and the number of Nissl bodies increased; SOD activity increased (P < 0.05) and MDA content decreased (P < 0.05); the expression of Nrf2, Keap1, NQO1 and HO-1 increased (P < 0.05) in the trillin group. Conclusion:Trillin may play a protective role in spinal cord injury by inhibiting oxidative stress response and improving the motor function.

OBJECTIVETo study the protective effect of Shenfu (Chinese traditional medicine) injections on myocardial ischemia/reperfusion (I/R) injury in rabbits.

METHODSThirty rabbits were divided into three groups (n = 10) randomly: control group, myocardial ischemia/ reperfusion group (MI/RI) and Shenfu injections extract group, three groups of rabbits were fed respectively with standard diet. After giving medicine 10 minutes, the myocardial ischemia/reperfusion injury animal model was established by ligaturing rabbits left ventricutar branch of coronary artery, and observing the changes of enzyme/hemodynamics during myocardial ischemia/reperfusion.

RESULTSIn model group myocardial function of shrink went down, the amount of malondialdehyde (MDA) was higher, the activity of superoxide dismutase (SOD), glutathine peroxidease (GSH-Px), Na(+) -K(+) -ATP and Ca(2+) -ATP were lower, lactic dehydrogenase (LDH) and creatine kinase(CK) were released compared with those in model group. Shenfu injections could recover left ventricular systolic pressure (LVSP) and +/- dp/dt(max), decrease left ventricular end-diastolic pressure(LVEDP), inhibit the increasing of MDA, LDH and CK, and increase the activaty of SOD, GSH-PX, Na(+) -K(+) -ATP and Ca(2+) -ATP.

CONCLUSIONShenfu injections can obviously protect myocardial ischemia/reperfusion injury.

Animals ; Calcium-Transporting ATPases ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Glutathione Peroxidase ; metabolism ; Hemodynamics ; drug effects ; Male ; Myocardial Ischemia ; physiopathology ; Myocardial Reperfusion Injury ; prevention & control ; Myocardium ; enzymology ; Rabbits ; Random Allocation ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Superoxide Dismutase ; metabolism

OBJECTIVETo study the effects of Enshi green tea tea polysaccharide on serum glucose in experimental diabetic rats.

METHODSDiabetic rats model were established by alloxan, Enshi green tea tea polysaccharide was poured into rats' stomach for four weeks, then the changes of the level about fasting blood glucose (FBG), glucokinase (GK), insulin (INS), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), spleen index and thymus index were observed.

RESULTSEnshi green tea tea polysaccharide could reduce the level of FBG in diabetic rats, and increase the activity of KG, SOD and GSH-Px, moreover, it could reduce the level of MDA and increase the spleen index and thymus index.

CONCLUSIONEnshi green tea tea polysaccharide has remarkable effect on playing down the blood sugar, and can increase the antioxygenic activity and immunity.

Animals ; Antioxidants ; pharmacology ; Blood Glucose ; metabolism ; Diabetes Mellitus, Experimental ; blood ; Male ; Oxidative Stress ; Plant Extracts ; pharmacology ; Polysaccharides ; pharmacology ; Rats ; Rats, Wistar ; Tea ; chemistry

OBJECTIVETo study the effects of the Rich Selenium-Banqiao-Codonopsis Pilosula (BCPA) injecta on the aged rats' immune functions and its underlying mechanism.

METHODSTotally 60 rats, composed of 2, 12 and 22 month age old (half male and half female), were served as a young group, middle-age group and aged group respectively. Each group rats were randomly divided into the control and the BCPA subgroup (n = 10). The BCPA group was injected with BCPA at 7.2 g/kg intraperitoneally every day and the control group was injected the same volume of normal saline. All rats were conventionally fed for 45 days. An immune injection was performed after 15 days of BCPA injection. On the 22nd day, late-onset immune response would be induced. The caudal vein blood was collected and the antigen specific IgG, IgG1 and IgG2a antibody was detected on the 15th, 30th and 45th day. On the 45th day, the major T cell subgroups of splenic cells were analyzed and splenic cells were proliferated.

RESULTSNo significant difference in the delayed-type hypersensivity (DTH) reaction was found between the control and the BCPA subgroups in the young and middle-aged rats while the aged BCPA subgroup had a stronger DTH reaction. There was no significant difference in the blood content of specific IgG, IgG1 and IgG2a antibody between the young and middle-age BCPA group while the aged BCPA group rats had an obvious enhancing reaction to the three antibodies mentioned above (P < 0.05). There was no obvious difference in the number of the CD3+ lymphocytes and the CD4+ T helper lymphocytes between the control and the BCPA subgroup in the young aged rats while a significant increase was spotted between the middle-aged and the aged group (P < 0.05). The splenic cells from young BCPA group rats had a strong proliferation response (P < 0.05).

CONCLUSIONBCPA can enhance DTH reaction, potentiate the production of specific IgG, IgG1 and IgG2a antibody to resist KLH, improve the reaction to antigen, increase the amount of CD4+ cell, promote the immune response and had an important role in anti-immunosenescence and antioxidant capacity improvement in the aged rats.

Aging ; Animals ; CD4-Positive T-Lymphocytes ; immunology ; Codonopsis ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Female ; Immune System ; drug effects ; Immunoglobulin G ; blood ; Male ; Rats ; Selenium ; pharmacology ; Spleen ; immunology