Hedysari Radix Polysaccharide Regulates Apoptosis of Smooth Muscle Cells in Gastric Antrum of Rat Model of Diabetic Gastroparesis via IGF-1/PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo observe the effects of Hedysari Radix polysaccharide on the apoptosis of gastric sinus smooth muscle cells and explore the underlying mechanism via the insulin-like growth factor-1 （IGF-1）/phosphatidylinositol 3-kinase （PI3K）/serine-threonine kinase （Akt） pathway in the rat model of diabetic gastroparesis （DGP）. MethodSixty-two Wistar male rats were randomized into a blank group （n=12） and a modelling group （n=50）. The rat model of DGP was established by small-dose multiple intraperitoneal injections of streptozotocin combined with an irregular high-fat and high-sugar diet for 4 weeks. The modeled rats were randomized into model group， mosapride citrate （1.35 mg·kg-1）， and high-， medium-， and low-dose （200， 100， and 50 mg·kg-1， respectively） Hedysari Radix polysaccharide groups. The rats were administrated with corresponding drugs by gavage， and those in the blank and model groups with equal volumes of pure water by gavage once a day for 8 consecutive weeks. The random blood glucose and body mass were measured every 2 weeks， and gastric emptying rate was calculated. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of smooth muscle in gastric antrum， and terminal deoxynucleoitidyl transferase-mediated nick-end labeling （TUNEL） was employed to detect the apoptosis of smooth muscle cells in the gastric antrum. The expression of IGF-1， phosphorylated （p）-PI3K， and p-Akt in the smooth muscle of gastric sinus tissue was detected by immunohistochemistry. Western blot was employed to determine the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） in the smooth muscle of the gastric antrum. ResultCompared with the blank group， the model group showed elevated random blood glucose at all time points （P<0.01）， decreased body mass and gastric emptying rate （P<0.01）， increased apoptotic index of smooth muscle cells in the gastric antrum （P<0.01）， down-regulated protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and up-regulated protein level of Bax （P<0.01）. Compared with the model group， the 8 weeks of drug administration lowered the random blood glucose， increased the body mass and gastric emptying rate （P<0.05， P<0.01）， decreased the apoptotic index of smooth muscle cells in the gastric antrum （P<0.05， P<0.01）， up-regulated the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and down-regulated the protein level of Bax （P<0.05， P<0.01）. Compared with the mosapride citrate group，the administration of low-dose Hedysari Radix polysaccharide for 6 and 8 weeks lowered the random blood glucose and decreased the body mass （P<0.05， P<0.01），low and medium-dose Hedysari Radix polysaccharide decreased the gastric emptying rate and the apoptotic index of smooth muscle cells in the astragaloside low-dose group decreased （P<0.05）. The protein levels of IGF-1，p-PI3K/PI3K，p-Akt/Akt and Bcl-2（low dose）were down-regulated and the protein level of Bax was up-regulated by low doses of Hedysari Radix polysaccharide （P<0.05， P<0.01）. Compared with high-dose Hedysari Radix polysaccharide， low-dose Hedysari Radix polysaccharide elevated random blood glucose and reduced body mass after 6 and 8 weeks of administration （P<0.05， P<0.01）， and the low and medium doses decreased the gastric emptying rate， increased the apoptotic index of smooth muscle cells in the gastric antrum （P<0.05， P<0.01）， down-regulated the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and up-regulated the protein level of Bax （P<0.05， P<0.01）. Compared with the medium-dose group，the low-dose group of Hedysari Radix polysaccharide had lower body mass，lower gastric emptying rate in rats，higher apoptotic index of smooth muscle cells in gastric sinus tissue after 6 and 8 weeks of administration （P<0.05， P<0.01）， and lower protein expression of IGF-1，p-PI3K/PI3K，p-Akt/Akt. ConclusionHedysari Radix polysaccharide protects the smooth muscle cells in gastric antrum against apoptotic injury and promotes gastric motility by activating the IGF-1/PI3K/Akt signaling pathway， as manifested by the up-regulated expression of IGF-1， p-PI3K， p-Akt， and Bcl-2 and down-regulated expression of Bax.

Dissecting the novel abilities of aripiprazole: The generation of anti-colorectal cancer effects by targeting Gαq via HTR2B

Colorectal cancer (CRC) is a type of malignant tumor that seriously threatens human health and life, and its treatment has always been a difficulty and hotspot in research. Herein, this study for the first time reports that antipsychotic aripiprazole (Ari) against the proliferation of CRC cells both in vitro and in vivo, but with less damage in normal colon cells. Mechanistically, the results showed that 5-hydroxytryptamine 2B receptor (HTR2B) and its coupling protein G protein subunit alpha q (Gαq) were highly distributed in CRC cells. Ari had a strong affinity with HTR2B and inhibited HTR2B downstream signaling. Blockade of HTR2B signaling suppressed the growth of CRC cells, but HTR2B was not found to have independent anticancer activity. Interestingly, the binding of Gαq to HTR2B was decreased after Ari treatment. Knockdown of Gαq not only restricted CRC cell growth, but also directly affected the anti-CRC efficacy of Ari. Moreover, an interaction between Ari and Gαq was found in that the mutation at amino acid 190 of Gαq reduced the efficacy of Ari. Thus, these results confirm that Gαq coupled to HTR2B was a potential target of Ari in mediating CRC proliferation. Collectively, this study provides a novel effective strategy for CRC therapy and favorable evidence for promoting Ari as an anticancer agent.

Effect of pre-injection of young rat plasma on cognitive dysfunction after cerebral ischemia-reperfusion in aged rats and the role of PI3K/Akt signaling pathway / 中华麻醉学杂志

Objective:To evaluate the effect of pre-injection of young rat plasma on cognitive dysfunction after cerebral ischemia-reperfusion (I/R) in aged rats and the role of phosphatidylinositol 3-kinase/serine threonine protein kinase (PI3K/Akt) signaling pathway.Methods:Seventy-two SPF-grade healthy male Sprague-Dawley rats, aged 18 months, weighing 600-650 g, were divided into 4 groups ( n=18 each) by the random number table method: control group (group C), cerebral I/R group (group IR), pre-injection of young rat plasma group (group P) and PI3K inhibitor LY294002 group (group LY). In group P and group LY, young rat plasma 100 μl/time was injected via the tail vein. In group C and group IR, the equal volume of normal saline was injected via the the tail vein, 2 times a week for 4 weeks. Then the model of cerebral I/R injury was developed under sevoflurane anesthesia in IR, P and LY groups. LY294002 0.3 mg/kg was injected through the tail vein at 1 h before anesthesia in LY group. The neurological deficit score (Longa score) was performed at 24 h after reperfusion, and then 6 rats were randomly sacrificed, and brain tissues were obtained to determine the cerebral infarct volume. Spontaneous mobility and anxiety-like behavior were assessed by the open field test at day 29 of reperfusion, and cognitive function was assessed by the novel object recognition test at day 30 of reperfusion. At the end of the behavioral test, rats were sacrificed, hippocampal tissues were isolated for determination of the expression of phosphorylated PI3K (p-PI3K), phosphorylated Akt (p-Akt), postsynaptic dense protein-95 (PSD-95) and synaptic vesicle protein (SYN) (by Western blot), and the dendritic length and dendritic spine density of neurons in the hippocampal CA1 region. Results:There was no significant difference in motor speed, distance traveled, and time of staying at the center of the open field among the four groups ( P>0.05). Compared with group C, the Longa score and cerebral infarct volume were significantly increased, the percentage of novel object exploration and discrimination index were decreased, the expression of p-PI3K, p-Akt, PSD-95 and SYN in hippocampal tissues was down-regulated, and the dendritic length and dendritic spine density of hippocampal neurons were decreased in IR, P and LY groups ( P<0.05). Compared with group IR, Longa score and cerebral infarct volume were significantly decreased, the percentage of novel object exploration and discrimination index were increased, the expression of p-PI3K, p-Akt, PSD-95 and SYN in hippocampal tissues was up-regulated, and the dendritic length and dendritic spine density of hippocampal neurons were increased in group P ( P<0.05), and no significant change was found in the parameters mentioned above in group LY ( P>0.05). Compared with group P, Longa score and cerebral infarct volume were significantly increased, the percentage of novel object exploration and discrimination index were decreased, the expression of p-PI3K, p-Akt, PSD-95 and SYN in hippocampal tissues was down-regulated, and the dendritic length and dendritic spine density of hippocampal neurons were decreased in group LY ( P<0.05). Conclusions:Pre-injection of young rat plasma can attenuate cognitive dysfunction after cerebral I/R in aged rats, and the mechanism is related to activation of hippocampal PI3K/Akt signaling pathway and improvement in synaptic plasticity.

Effect of parathyroid hormone on osteoporosis after spinal cord injury in rats and its mechanism / 中国药理学通报

Aim To explore the effect of parathyroid hormone on osteoporosis in rats after spinal cord injury(SCI)and its mechanism.Methods SD rats were divided into sham operation group(Sham), SCI model group(SCI), SCI+parathyroid hormone group(SCI+PTH)and SCI+PTH+transfected miR-146a irrelevant fragment group(SCI+PTH+NC)and SCI+PTH+transfection miR-146a inhibitor group(SCI+PTH+miR-146a inhibtor), and then given 60 μg·kg-1 PTH(SCI+PTH group), 60 μg·kg-1 PTH and 20 pm miR-146a NC(SCI+PTH+NC group)or 60 μg·kg-1 PTH and 20 pm miR-146a inhibitor(SCI+PTH+miR-146a inhibitor group)by tail vein injection every 3 d for 8 weeks.Rats in Sham group and SCI group were given equal amount of saline in the same way.The behavioral movement scores of rats were recorded by the BBB scoring method 1 d, 7 d, 14 d, 21 d, 28 d, and 56 d after operation; serum calcium(Ca)and alkaline phosphatase(ALP)were measured using the kits; bone mineral density of femur and tibia was measured by a bone mineral density scanner; the morphological changes of rat spinal cord were observed by HE staining; expression of miR-146a was detected by qRT-PCR and protein expression of p-PI3K and p-Akt was detected by Western blot.Results Compared with Sham group, SCI group had decreased BBB score(P<0.05 or P<0.01), serum Ca, femoral and tibial bone mineral density content and expression of miR-146a, p-PI3K and p-Akt, but increased serum ALP(P<0.01).Compared with SCI group, BBB score(P<0.05 or P<0.01), serum Ca, femoral and tibia bone mineral density content, and the expression of miR-146a, p-PI3K and p-Akt( P<0.01)increased, together with decreased serum ALP in SCI+PTH group(P<0.01).Compared with SCI+ PTH group, the above indicators of rats were significantly inhibited in SCI+PTH+miR-146a inhibitor group.Conclusions PTH has certain therapeutic effect on SCI osteoporosis, achieved possibly by regulating miR-146a/PI3K/Akt signaling.

Therapeutic regulation of autophagy in hepatic metabolism

Metabolic homeostasis requires dynamic catabolic and anabolic processes. Autophagy, an intracellular lysosomal degradative pathway, can rewire cellular metabolism linking catabolic to anabolic processes and thus sustain homeostasis. This is especially relevant in the liver, a key metabolic organ that governs body energy metabolism. Autophagy's role in hepatic energy regulation has just begun to emerge and autophagy seems to have a much broader impact than what has been appreciated in the field. Though classically known for selective or bulk degradation of cellular components or energy-dense macromolecules, emerging evidence indicates autophagy selectively regulates various signaling proteins to directly impact the expression levels of metabolic enzymes or their upstream regulators. Hence, we review three specific mechanisms by which autophagy can regulate metabolism: A) nutrient regeneration, B) quality control of organelles, and C) signaling protein regulation. The plasticity of the autophagic function is unraveling a new therapeutic approach. Thus, we will also discuss the potential translation of promising preclinical data on autophagy modulation into therapeutic strategies that can be used in the clinic to treat common metabolic disorders.

Role of AMPK/p38 MAPK/Nrf2 signaling pathway in myocardial ischemia-reperfusion injury in diabetic rats / 中华麻醉学杂志

Objective:To evaluate the role of adenosine monophosphate-dependent protein kinase/p38 mitogen-activated protein kinase/nuclear factor E2-associated factor 2 (AMPK/p38 MAPK/Nrf2) pathway in myocardial ischemia-reperfusion (I/R) injury in diabetic rats.Methods:Clean-grade healthy Sprague-Dawley male rats, aged 2-3 months, weighing 220-280 g, were fed with a high fat diet, and 1% streptozotocin 50 mg/kg was intraperitoneally injected for 4 consecutive days to develop the model of diabetes mellitus.Thirty diabetic rats were divided into 3 groups ( n=10 each) using the random number table method: sham operation group (sham group), myocardial I/R group (I/R group), and AMPK inhibitor compound C+ myocardial I/R group (C+ I/R group). The model of myocardial I/R injury was developed by ligation of the left anterior descending coronary artery for 30 min followed by 120 min reperfusion.Compound C 0.5 mg/kg was injected via the caudal vein at 30 min before ischemia in C+ I/R group, while the equal volume of normal saline was given instead in Sham group and I/R group.At 120 min of reperfusion, the percentage of myocardial infarct size was calculated, the serum concentrations of creatine kinase isoenzymes (CK-MB) and lactic dehydrogenase (LDH) were determined by enzyme-linked immunosorbent assay, the levels of glutathione (GSH), superoxide dismutase (SOD) and reactive oxygen species (ROS) in myocardial tissues were measured by enzyme-linked immunosorbent assay, and the expression of AMPK, phosphorylated AMPK (p-AMPK), phosphorylated p38 MAPK (p-p38 MAPK), Nrf2 and heme oxygenase-1 (HO-1) in myocardium was determined by Western blot. Results:Compared with Sham group, the percentage of myocardial infarct size and serum CK-MB and LDH levels were significantly increased, the levels of GSH and SOD in myocardial tissues were decreased, ROS level was increased, and the expression of AMPK, p-AMPK, p-p38 MAPK, Nrf2 and HO-1 was up-regulated in I/R group ( P<0.05). Compared with I/R group, the percentage of myocardial infarct size and serum CK-MB and LDH levels were significantly increased, the levels of GSH and SOD in myocardial tissues were decreased, ROS level was increased, and the expression of AMPK, Nrf2 and HO-1 was down-regulated in C+ I/R group ( P<0.05). Conclusions:AMPK/p38 MAPK/Nrf2 signaling pathway is involved in the mechanism of endogenous antioxidant stress during myocardial I/R in diabetic rats.

The Effect of Doxycycline on the Expression of MMP-2 and MMP-9 in Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To investigate the expression of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in multiple myeloma (MM) patients, and analyze the effect of doxycycline (DOX) on the expression of MMP-2 and MMP-9 in MM cells.@*METHODS@#The peripheral blood and bone marrow samples of MM patients were collected, and the patients were divided into three groups: newly diagnosed group, remission group and relapsed/refractory group, while the peripheral blood samples of 34 health people and the bone marrow samples of 17 IDA patients were selected as normal control and control group. The levels of MMP-2 and MMP-9 were detected by ELISA. The protein levels of MMP-2 and MMP-9 in H929 cells treated by different concentrations of DOX were analyzed by Western blot. After H929 cells was treated by Akt inhibitor MK-2206 2HCl in combination with DOX, Western blot was used to detect the levels of MMP-2 and MMP-9.@*RESULTS@#The levels of MMP-2 and MMP-9 in newly diagnosed MM patients were higher than those in control (P<0.05), while for the patients in the remission group were decreased, but still higher than those in control. The levels of MMP-2 and MMP-9 were increased again for the patients in relapsed/refractory group, and showed no significant difference as compared with those in newly diagnosed group. The levels of MMP-2 and MMP-9 could be inhibited by 10 mg/L and 15 mg/L DOX treated by H929 cell. The protein levels of MMP-2 and MMP-9 showed no altered in H929 cells treated by 5 nmol/L MK-2206 2HCl alone. DOX exerted more profound inhibitory effect to MMP-2 and MMP-9 expression in H929 cells when Akt inhibitor MK-2206 2HCl was combined with DOX.@*CONCLUSION@#The levels of MMP-2 and MMP-9 are increased in MM patients and related to the disease status of MM. DOX can inhibit the expression of MMP-2 and MMP-9 in MM cells, and antagonizing its activation of Akt signaling pathway can further enhance the inhibitory effect.

Effect of Sanshi Shengxin Ointment on healing of pressure ulcers in rats based on PD K/Akt signal axis / 中国药理学通报

Aim To explore the effect of Sanshi Shengxin Ointment on the healing of pressure wounds in rats and its mechanism. Methods The experimental animals were divided into sham operation group (Sham), model group (Model), Bei Fuxin group (bFGF), Sanshi Shengxin ointment group (TM), and Sanshi Shengxin ointmen + PI3K blocker group (TM + LY294002). The pressure ulcer rat model was prepared by the local tissue ischemia-reperfusion injury method. The wound healing rate of rats in each group was observed on 3rd, 7th, and 14th day. On 3rd day of the dressing change, the pathological changes of the wound, the plasma VEGF content, and the expression of VEGF and PI3 K/Akt signal-related proteins in the wound skin tissue were detected. Results Compared with sham group, the wound healing rate of rats in model group was reduced (P < 0. 01) with severe wound injury. The plasma VEGF content decreased (P <0. 01), and the expression of VEGF, p-PI3K, and p-Akt decreased too (P <0. 01). Compared with model group, the wound healing rate and VEGF content of rats in TM group increased (P < 0. 05 or P < 0. 01), the wound injury was significantly improved, and the expression of VEGF, p-PI3K, and p-Akt increased (P <0. 05 or P < 0. 01). Compared with TM group, the wound healing rate and the content of VEGF decreased in TM + LY294002 rats (P <0. 01), the wound injury was aggravated, and the expressions of tissue VEGF, P-PI3K, and p-Akt decreased (P < 0.05 or P < 0.01). Conclusions Sanshi Shengxin Ointment can up-regulate the expression of VEGF through PI3K/Akt signal and promote the healing of pressure sore in rats.

Hydromorphone postconditioning attenuates myocardial ischemia/reperfusion-induced apoptosis in rats through PI3K/AKT pathway / 中华急诊医学杂志

Objective:To investigate the role of PI3K/Akt signaling pathway in hydromorphone postconditioning on alleviating myocardial ischemia/reperfusion (I/R)-induced apoptosis in rats.Methods:Forty healthy male SD rats were randomly(random number) divided into five groups, with 8 rats in each group:①sham group;②I/R group;③I/R+hydromorphone group (I/R+H group);④I/R+PI3K inhibitor group (I/R+W group); and⑤I/R+hydromorphone+PI3K inhibitor group (I/R+H+W group). The myocardial ischemia/reperfusion injury model was established by ligating the left anterior descending coronary artery for 30 min and reperfusion for 120 min. After the experiment, the area of myocardial infarction was measured by 2, 3, 5-triphenyl tetrazolium chloride (TTC) staining. The amount of serum lactate dehydrogenase (LDH) leakage was estimated by colorimetry . The cardiomyocyte apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling (TUNEL) assay. The protein expressions of p-Akt, Bcl-2 and Bax were detected by Western blot. Comparisons among groups were carried out by analysis of variance (ANOVA).Results:Compared with the sham group, the area of myocardial infarction, serum LDH leakage and cardiomyocyte apoptosis were significantly increased, p-Akt and Bax expression were upregulated, Bcl-2 expression was downregulated in the I/R group ( P<0.05). Compared with the I/R group, the area of myocardial infarction, serum LDH leakage and cardiomyocyte apoptosis were markedly decreased, p-Akt and Bcl-2 expression were upregulated and Bax expression was downregulated in the I/R+H group ( P<0.05). Compared with the I/R+H group, the area of myocardial infarction, serum LDH leakage and cardiomyocyte apoptosis were significantly increased, p-Akt and Bcl-2 expression were downregulated, and Bax expression was upregulated in the I/R+H+W group ( P<0.05). Conclusions:Hydromorphone postconditioning can alleviate cardiomyocyte apoptosis induced by myocardial ischemia/reperfusion, and its protection mechanism may be related to the activation of PI3K/Akt signaling pathway.

A Analysis on Expression Significance of BUB1 in Breast Cancer and Intervention of Japonicone A Based on Data-mining from Bioinformatics / 中国实验方剂学杂志

Objective:To investigate the expression of the cell cycle mitotic spindle checkpoint serinel/threonine kinase （BUB1） gene in breast cancer and its relationship with the prognosis，and further explore the intervention effect of japonicone A on BUB1 gene in breast cancer cells. Method:Through Oncomine，GEPIA，GEO database，bc-GenExMiner v4.5 and Kaplan-Meier Plotter database， in-depth mining was conducted for BUB1 gene expression-related data，in order to explore the difference in the expression of BUB1 gene in breast cancer tissues and normal breast tissues and its relationship with patient prognosis. Encyclopedia of Cancer Cell Lines （CCLE） was used to analyze the expression of BUB1 in T cells and B cells of breast cancer tissues，STRING database was used to draw BUB1-related protein network diagram and gene ontology（GO） function annotation and analyze relevant pathways in Kyoto Encyclopedia of Genes and Genomes （KEGG）. The tumor immune assessment resource（TIMER） database was used to analyze the expression of BUB1 in immune infiltrates and its impact on the survival and prognosis of patients with gastric cancer. Finally， the effect of japonicone A on the expression level of BUB1 gene in breast cancer cells was further analyzed. Result:①The mRNA level of BUB1 in breast cancer tissues was significantly higher than that of normal tissue samples， but the expression levels of BUB1 in breast cancer patients with different molecular subtypes varied. ② Increased expression of BUB1 could lead to longer distant metastasis-free survival（DMFS），overall survival（OS），and recurrence free survival（RFS） in luminal A subtypes. ③ BUB1 was positively correlated with structural maintenance of chro-mosome 4（SMC4） mRNA expression level， and might be interacted with 10 proteins， such as NUF2， with the strongest interaction relationship. ④ The high expression of BUB1 mRNA in CD8⁺T cells and neutrophils in breast invasive carcinoma（BRCA） and BRCA-basal had a better two-year survival prognosis than the low expression group；the low expression of BUB1 mRNA in B cells in BRCA-Her2 had a better two-year survival prognosis than the high expression group. ⑤ GEO database analysis data set GSE85871 found that japonicone A could down-regulate the expression of BUB1 gene in breast cancer MCF7 cells. Conclusion:BUB1 gene is highly expressed in breast cancer tissues and related to the prognosis of breast cancer and immune cell infiltration. This may be a new potential therapeutic target for japonicone A to intervene breast cancer cells.

Effect of Shuangshen Xionglian Granule on Vasodilation and PI3K/Akt/eNOS Pathway in Hyperhomocysteinemia Chronic Kidney Disease Rats / 中国实验方剂学杂志

Objective:To study the effect of Shuangshen Xionglian （SSXL） granules on vasculopathy and phosphatidylinositol 3 kinase （PI3K）/serine threonine kinase （Akt）/nitrogen oxide synthase （eNOS） signal in hyperhomocysteinemia chronic kidney disease rats. Method:Rats were randomly divided into 5 groups： sham operation group， model group， and high， medium and low-dose （8， 4， 2 g·kg-1） SSXL groups. The model of hyperhomocysteinemia chronic kidney disease in rats was established with high methionine feed combined with 5/6 nephrectomy. After 5/6 nephrectomy， continuous intragastric administration lasted for four weeks. Arterial blood pressure was measured at the 4th and 8th weeks after operation. At the end of the 8th week after the operation， blood was collected to determine serum creatinine， urea nitrogen， homocysteine （Hcy）， methionine and blood lipid. Western blot was used to detect the expressions of PI3K/Akt/eNOS pathway-related proteins， such as p-p85， p-Akt and p-Ser177 in thoracic aorta， and serum NO and eNOS were measured. The changes of endothelium-dependent relaxation and non-endothelium-dependent relaxation were measured by the method of isolated thoracic aorta ring. Pathological htoxylin eosin （HE） staining was used to observe the changes of renal tissue and thoracic aorta. Result:At the 8th week of the experiment， compared with the sham operation group， arterial systolic blood pressure， serum urea nitrogen， creatinine， Hcy， methionine， total cholesterol and low-density lipoprotein of the model group were significantly increased. Four weeks later after administration， arterial systolic blood pressure， serum urea nitrogen， Hcy， methionine， serum total cholesterol and serum low-density lipoprotein were significantly reduced in each dose group （P<0.05， P<0.01）. The creatinine in the SSXL 8， 4 g·kg-1 group was significantly reduced （P<0.05）. The nitric oxide content of SSXL in each dose groups were increased compared with that in the model group （P<0.05， P<0.01）， and the serum eNOS activity of the SSXL in the SSXL 8 g·kg-1 group was significantly increased compared with that in the model group （P<0.05）. The endothelium dependent and non-endothelium dependent vasodilation of thoracic aortic rings in the model group were significantly damaged. The cumulative concentration of acetylcholine （1×10-5.5~1×10-4 mmo1·L-1） in the SSXL 8 g·kg-1 group was significantly improved （P<0.05， P<0.01）. The diastolic degree of the vascular ring in the SSXL 8 g·kg-1 group was significantly higher than that in the model group （P<0.05）. Western blot results showed that the expressions of p-85， p-Akt and p-Ser177 in blood vessels increased in the sham group compared with those in the model group （P<0.01）. Compared with the model group， the phosphorylation level of this pathway was increased in the SSXL groups， and the expressions of p-Akt and p-Ser177 in the SSXL 8 g·kg-1 group were significantly increased （P<0.05）. The pathological results showed that the pathological changes of thoracic aorta and renal tissue in the dosages of SSXL were significantly reduced compared with those in the model group. Conclusion:SSXL granules can improve hyperhomocysteine and dyslipidemia in rats of chronic kidney disease with hyperhomocysteine， reduce serum creatinine， urea nitrogen levels and arterial systolic blood pressure， and improve vascular morphology and diastolic function， which may be related to the regulation of the PI3K/Akt/eNOS signaling pathway.

Phosphorylation regulates cullin-based ubiquitination in tumorigenesis

Cullin-RING ligases (CRLs) recognize and interact with substrates for ubiquitination and degradation, and can be targeted for disease treatment when the abnormal expression of substrates involves pathologic processes. Phosphorylation, either of substrates or receptors of CRLs, can alter their interaction. Phosphorylation-dependent ubiquitination and proteasome degradation influence various cellular processes and can contribute to the occurrence of various diseases, most often tumorigenesis. These processes have the potential to be used for tumor intervention through the regulation of the activities of related kinases, along with the regulation of the stability of specific oncoproteins and tumor suppressors. This review describes the mechanisms and biological functions of crosstalk between phosphorylation and ubiquitination, and most importantly its influence on tumorigenesis, to provide new directions and strategies for tumor therapy.

Effect of pre-infusion of young rat plasma on postoperative cognitive function in aged rats and role of PI3K/Akt signaling pathway / 中华麻醉学杂志

Objective:To evaluate the effect of pre-infusion of young rat plasma on postoperative cognitive function in aged rats and role of phosphatidylinositol 3 kinase/serine-threonine protein kinase (PI3K/Akt) signaling pathway.Methods:Sixty SPF healthy male Sprague-Dawley rats, aged 18 months, weighing 550-650 g, were divided into 4 groups ( n=15 each) using a random number table method: control group (group C), operation group (group O), young rat plasma group (group P) and PI3K inhibitor LY294002 group (group LY). The young rat plasma 100 μl/time was injected via the caudal vein twice a week for 4 consecutive weeks in group P and group LY, while the equal volume of normal saline was given instead in group C and group O. Rats received internal fixation for unilateral tibial fracture under sevoflurane anesthesia in O, P and LY groups.Rats received no treatment in group C. PI3K inhibitor LY294002 0.3 mg/kg was injected through the caudal vein before anesthesia in group LY.The ability of spontaneous activity was evaluated by open field test at 3 days after surgery, and then the cognitive function was assessed by Morris water maze test.The rats were sacrificed after the end of behavioral testing, and the hippocampal tissues were isolated for determination of the expression of phosphorylated PI3K (p-PI3K), phosphorylated Akt (p-Akt), synapsin, synaptophysin I and synaptic vesicle protein (by Western blot) and for microscopic examination of the ultrastructure of hippocampal neurons (with a transmission electron microscope). The number of synapses was recorded. Results:There was no significant difference in the movement speed and length and time spent in the central zone among the four groups ( P>0.05). Compared with group C, the escape latency was significantly prolonged, the number of crossing the original platform was reduced, the expression of p-PI3K, p-Akt, synapsin, synaptophysin I and synaptic vesicle protein was down-regulated, and the number of synapses was reduced in O and LY groups ( P<0.05), and no significant change was found in the parameters mentioned above in group P ( P>0.05). Compared with group O, the escape latency was significantly shortened, the number of crossing the original platform was increased, the expression of p-PI3K, p-Akt, synapsin, synaptophysin I and synaptic vesicle protein was up-regulated, and the number of synapses was increased in group P ( P<0.05), and no significant change was found in the parameters mentioned above in group LY ( P>0.05). Compared with group P, the escape latency was significantly prolonged, the number of crossing the original platform was reduced, the expression of p-PI3K, p-Akt, synapsin, synaptophysin I and synaptic vesicle protein was down-regulated, and the number of synapses was reduced in group LY ( P<0.05). Conclusion:Pre-infusion of young rat plasma can improve postoperative cognitive function in aged rats, and the mechanism is related to activation of PI3K/Akt pathway and improvement of synaptic plasticity.

Effects of silencing Kat5 gene on proliferation and apoptosis of thyroid papillary carcinoma cells / 吉林大学学报(医学版)

Objective: To investigate the effects of lysine acetyltransferase (Kat5) on the proliferation and apoptosis of thyroid papillary carcinoma cells. Methods: RT-PCR and Western blotting methods were used to determine the expression levels of Kat5 mRNA and protein in the thyroid papillary carcinoma BHP10-3. TPC-1 . K1 cells and thyroid epithelial Nthy-ori3-l cells. The thyroid papillary carcinoma TPC-1 cells were divided into blank control group, negative control group and silencing Kat5 group (si-Kat5 group). The cells in blank control group were not transfected. the cells in negative control group were transfected with negative control siRNA. and the cells in si-Kat5 group were transfected with Kat5 siRNA. The expression levels of Kat5 mRNA and protein in the thyroid papillary carcinoma TPC-1 cells in various groups were detected by RT-PCR and Western blotting methods. The proliferation activities of the TPC-1 cells in various groups were measured by CCK-8 method, the colony formation assay was used to measure the clone formation rates of the TPC-1 cells in various groups, the apoptotic rates of the TPC-1 cells in various groups were detected by flow cytometry, and the expression levels of cyclin-dependent kinase 2 (CDK2). P21, P53. cleaved caspase-3. P13K, phosphorylated P13K (p-P13K), AKT, and phosphorylated AKT (p-AKT) protein in the TPC-1 cells in various groups were detected by Western blotting method. Results: The expression levels of Kat5 mRNA and protein in the thyroid papillary carcinoma BHP10-3. TPC-1 and K1 cells were higher than those in thyroid epithelial Nthy-ori3-l cells (P<0. 05). The differences in the Kat5 mRNA and protein expression levels, the proliferation activities, the clone formation rates, the apoptotic rates and the expression levels of CDK2. P21. P53. cleaved caspase-3. P13K. p-P13K . AKT and p-AKT proteins in the TPCM cells between various groups were statistically significant (P<0.05). Compared with blank control group and negative control group, the expression levels of Kat5 mRNA and protein in the TPC-1 cells in si-Kat5 group were decreased ( P

Research progress in immune checkpoint inhibitors in the treatment of oncogenedriven advanced nonsmall cell lung cancer / 中南大学学报(医学版)

The clinical application of immune checkpoint inhibitors (ICIs) lead to dramatic changes in the treatment strategy for patients with advanced non-small cell lung cancer (NSCLC), but the efficacy of ICIs in oncogene-driven NSCLC is controversial. Existing research shows that the efficacy of ICIs may be related to different types of driver genes, programmed cell death-ligand 1 (PD-L1) level, and tumor mutational burden (TMB). It may involved in other factors, such as clinical characteristics, and immune cell density. ICIs monotherapy or combination therapy may play a role in a subset of oncogene-driven NSCLC patients, but further studies are needed to select these patients, which may be an important direction for the future development of advanced NSCLC.

Miltirone induces cell death in hepatocellular carcinoma cell through GSDME-dependent pyroptosis

Pyroptosis is a form of programmed cell death, and recently described as a new molecular mechanism of chemotherapy drugs in the treatment of tumors. Miltirone, a derivative of phenanthrene-quinone isolated from the root of Bunge, has been shown to possess anti-cancer activities. Here, we found that miltirone inhibited the cell viability of either HepG2 or Hepa1-6 cells, and induced the proteolytic cleavage of gasdermin E (GSDME) in each hepatocellular carcinoma (HCC) cell line, with concomitant cleavage of caspase 3. Knocking out switched miltirone-induced cell death from pyroptosis to apoptosis. Additionally, the induction effects of miltirone on GSDME-dependent pyroptosis were attenuated by siRNA-mediated caspase three silencing and the specific caspase three inhibitor Z-DEVD-FMK, respectively. Miltirone effectively elicited intracellular accumulation of reactive oxygen species (ROS), and suppressed phosphorylation of mitogen-activated and extracellular signal-regulated kinase (MEK) and extracellular regulated protein kinases 1/2 (ERK1/2) for pyroptosis induction. Moreover, miltirone significantly inhibited tumor growth and induced pyroptosis in the Hepa1-6 mouse HCC syngeneic model. These results provide a new insight that miltirone is a potential therapeutic agent for the treatment of HCC GSDME-dependent pyroptosis.

Citron Rho-interacting serine/threonine kinase knockdown suppresses prostate cancer cell proliferation and metastasis by blocking Hippo-YAP pathway / 南方医科大学学报

OBJECTIVE@#Citron Rho-interacting serine/threonine kinase (CIT) was identified recently as an oncogene involved in the progression of various malignant tumors, but its role in prostate cancer (PCa) remains unclear. In this study, we aimed to investigate the biological functions of CIT in PCa.@*METHODS@#We analyzed the expression of CIT in PCa tissues and its clinical correlations based on the Cancer Genome Atlas (TCGA) and Memorial Sloan-Kettering Cancer Center (MSKCC) dataset. We then examined the effects of RNA interference-mediated CIT silencing on the proliferation, migration and invasion of PC-3 cells using cell counting kit-8, wound healing assay and Transwell assay. We also investigated the effect of CIT silencing on epithelial-mesenchymal transition (EMT) and Hippo-Yap signaling pathway in the cells using Western blotting.@*RESULTS@#CIT expression was significantly elevated in PCa tissues from TCGA cohort ( < 0.05). MSKCC dataset analysis showed that an elevated expression of CIT was significantly correlated with N stage (=0.001), distant metastasis ( < 0.001), Gleason score (=0.010) and PSA (=0.004). In cultured PC-3 cells, knockdown of CIT significantly inhibited cell proliferation, migration and invasion, reversed the EMT phenotype and decreased the expression and activity of YAP.@*CONCLUSIONS@#CIT might function as an oncogene in PCa by modulating the Hippo-YAP signaling pathway and serve as a candidate therapeutic target for PCa.

Effect of Yanghe Huayan Tang Combined with PI3K Inhibitor on Expressions of HER-2, PI3K and p-Akt in PI3K/Akt Pathway of High-expression HER-2 Breast Cancer Cell Lines / 中国实验方剂学杂志

Objective: To investigate effect of Yanghe Huayan Tang and phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 in intervening expressions of human epidermal growth factor receptor human epidermal growth factor receptor-2(HER-2), PI3K and phosphorylated serine/threonine kinase (p-Akt)in PI3K/Akt pathway of breast cancer cell line SK-BR-3 (HER-2 high expression) in vitro, and study intervention mechanism of Yanghe Huayan Tang. Method: Yanghe Huayan Tang intestinal absorption fluid was prepared, breast cancer cell strain SK-BR-3 was divided into blank group, Yanghe Huayan Tang group, LY294002 group, LY294002+Yanghe Huayan Tang group. The concentration was respectively 125 g·L-1 for Yanghe Huayan Tang, 0.05 g·L-1 for LY294002, 125 g·L-1 for Yanghe Huayan Tang+LY294002.The expressions of HER-2, PI3K and p-Akt protein were detected by immunohistochemistry and Western blot after 24 h. The most appropriate and effective concentration was obtained through extensive experiments. The expressions of HER-2, PI3K and p-Akt were detected by reverse transcription polymerase chain reaction (RT-PCR). Result: Compared with blank group, LY294002 group, and LY294002+Yanghe Huayan Tang group could inhibit protein and mRNA expressions of HER-2, PI3K, p-Akt in breast cancer cell lines(PPPPConclusion: Yanghe Huayan Tang can inhibit angiogenesis and invasion of breast cancer. Its main mechanism is expressed by intervening PI3K/Akt pathway, reducing expressions of HER-2, PI3K and p-Akt in pathway.

Expression of Pim-1 in primary hepatocellular carcinoma and its clinical significance / 实用肿瘤学杂志

Objective The aim of this study was to determine the expression of Pim-1 in primary hepatocellular carcinoma (PHC)and its clinical significance. Methods Immunohistochemical staining(IHC) was used to detect the expression of Pim-1 in 122 cases of PHC tissues,corresponding paracancer tissues,and 85 normal liver tissues. The relationship between the expression of Pim-1 protein and clinicopathological parameters was analyzed retrospectively. K-M method was used to analyze the effect of differ-ent Pim-1 expression on the survival time of PHC patients. Cox proportional hazard regression models were used to analyze risk fac-tors affecting survival time in patients with PHC. Results The total positive expression rate of Pim-1 protein in PHC tissues was 88. 5% ,which was significantly higher than that in adjacent tissues and normal liver tissues(P<0. 05). Univariate statistical analysis showed that patients with high expression of Pim-1 protein had poor preoperative liver function,more tumors,larger tumor diameter, high incidence of lymph node metastasis and high TNM stage(P<0. 05). Survival analysis suggested that the survival time of patients in the low expression group was significantly longer than that in the high expression of Pim-1 group(P<0. 05). Multivariate statisti-cal analysis showed that high expression of Pim-1 protein was an independent risk factor for survival time in patients with PHC(P<0. 001). Conclusion The expression level of Pim-1 in PHC tissues is significantly increased,which is related to the clinical pro-gress of PHC and the survival time of patients. Pim-1 overexpression indicates the poor prognosis of PHC patients.

Receptor interacting protein serine threonine kinase 3 mediates macrophage activation and interleukin-6 production in autoimmune hepatitis / 中华消化杂志

Objective To investigate the expression level of receptor-interacting protein serine threonine kinase (RIP) 3 in macrophages/monocytes activation in autoimmune hepatitis (AIH) and its regulation on inflammatory cytokines.Methods The degree of macrophage infiltration and the expression of RIP3 in liver tissues from patients with AIH or hepatic cysts by double-immunofluorescence.After 24 hours treated with different concentrations (0,1,3,6,10 μg/mL) of lipopolysaccharide (LPS),Necrostatin-1,the specific inhibitor of RIP3 signaling pathway,and 6-thioguanine,the active metabolite of azathioprine,the expression levels of RIP1 (RIP3 upstream signal molecule),RIP3 and mixed-lineage kinase domain-like protein (MLKL,RIP3 downstream substrate) in RAW264.7 macrophages were detected by Western blotting.The expression of macrophage-associated cytokine at mRNA level of each treatment group was determined by real time quantitative polymerase chain reaction (PCR).Student's t test and sum rank test were performed for statistical analysis.Spearman analysis was performed for the correlation analysis.Results Compared with hepatic cysts adjacent liver tissues,the infiltration of CD68 positive macrophages in liver tissue of AIH patients was significantly increased (4.75 ± 0.96 vs 28.86 ± 6.23),and the difference was statistically significant (t =7.80,P＜0.05),and the expression level of RIP3 also was significantly increased (15,11 to 22 vs 0,0 to 1),and the difference was statistically significant (Z=-2.66,P＜0.05).In vitro,compared with those of control group,the expression levels of RIP1,RIP3 and MLKL stimulated by LPS at 0,1,3,6,10 μg/mL were significantly increased,and the differences were statistically significant (t=4.00,4.90,6.40,10.30;3.80,9.30,9.80,9.00;4.90,9.90,9.30 and 7.70;all P＜0.05),and were dose-dependent (r=0.91,0.86 and 0.79,all P＜0.05).Furthermore,compared with those of LPS-stimulated group,the expressions of RIP1,RIP3,MLKL of LPS+Necrostatin-1 group were significantly decreased (0.73±0.11 vs 0.47±0.13,0.60±0.07 vs 0.37 ± 0.05,0.65 ± 0.22 vs 0.38 ± 0.04,respectively),and the differences were statistically significant (t=2.60,4.50 and 2.10,all P＜0.05).And the expressions of interleukin (IL)-1β,IL-6and IL-10 at mRNA levels were also decreased (810.3±200.8 vs 463.7±118.1,1 504.4±482.7 vs 290.4±106.9,1 358.6 ± 559.2 vs 677.8 ± 297.6,respectively),and the differences were statistically significant (t=5.40,12.52,5.70,all P＜0.05).However,the expressions of IL-4 and TGF-β at mRNA levels up-regulated (0.3±0.2 vs 0.6±0.3,0.4±0.1 vs 0.9±0.4,respectively),and the differences were statistically significant (t=4.60 and 6.10,both P＜0.05).Compared with those of the LPS-stimulated group,the expressions of IL-1β,IL-6 and IL-10 at mRNA levels of LPS and 6-thiopurine stimulated group significantly down-regulated (810.3±200.8 vs 283.4±65.5,1 504.4±482.7 vs 354.4±73.8,1 358.6± 559.2 vs 625.6±336.3),and the differences were statistically significant (t=4.30,10.60 and 3.50,all P＜0.05);however,the expressions of IL-4 and TGF-β at mRNA levels significantly up-regulated (0.3±0.2 vs 0.6±0.1 and 0.4±0.1 vs 0.5±0.1),and the differences were statistically significant (t=5.20and 12.50,P＜0.05).Conclusions The regulation effects of 6-thiopurine on RIP3 signaling pathway and related cytokines are similar to those of Necrostatin-1.And the expression of RIP3 signaling protein increasing in activated macrophages of liver tissues from AIH patients is closely related to the regulation of IL-6.The RIP3 mediated inflammatory signaling pathway in macrophage may be involved in the genesis and development of AIH and may be a potential therapeutic target.