Objective To investigate the impact of silymarin(SM)on the malignant growth of glioma cells and the regulatory mechanism on the miR-124-3p/WEE1 axis.Methods Glioma U87 cells were grouped into control,SM low,medium,and high concentration groups,and SM high concentration + miR-124-3p inhibitor group(SM high + miR-124-3p inhibitor group).CCK-8 was used to measure the proli-feration rate of cells;Transwell? assay was applied to assay the migration and invasion of cells;cell cycle progression was detected by flow cytometry;Western blotting was applied to measure the expression of cyclin D1 and apoptosis-related proteins;the levels of miR-124-3p and WEE1 mRNA were determined by qRT-PCR;and a luciferase activity test was applied to verify the targeting relationship between miR-124-3p and WEE1;in addition,the establishment,administration,and analysis of a NOD/SCID mouse model of intracranial trans-planted tumor were conducted.Results Compared with the control group,the cell proliferation,the numbers of migrating and invading cells,the expression of cyclin D1,and the level of WEE1 mRNA in the various SM treatment groups decreased,the number of cells in G0/G1 phase,the expression of cleaved caspase-8,cleaved caspase-9,cleaved caspase-3 and miR-124-3p increased(P＜0.05);furthermore,transfection of miR-124-3p inhibitor reversed the inhibitory effect of SM on the malignant behavior of glioma cells.In vivo experiments with mice showed that the weights and volumes of tumors in the SM treatment group were lower than those in the model group(P＜0.05),and there was no discernible change in the weight of the mice(P＞0.05).Conclusion SM can inhibit the malignant growth of glioma cells by upregulating miR-124-3p and downregulating WEE1.

Objective To investigate the therapeutic efficacy of dandelion extract on intracerebral hemorrhage(ICH)rats and its effect on nuclear factor erythroid 2 related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway.Methods Stereotaxic intracranial injection of type Ⅳ col-lagenase was used to establish rat ICH model.Then 48 ICH rats were randomly divided into mod-el group,dandelion extract group,Nrf2 inhibitor(ML385)group and dandelion extract+ML385 group,with 12 rats in each group.Another 12 rats served as sham operation group.After treat-ment,neurological deficits was evaluated and scored for all groups of rats.Blood-brain barrier(BBB)function,neuronal apoptotic rate in the hippocampus,serum levels of COX-2,IL-6 and iNOS,cerebral contents of CAT,GSH-Px,ROS and MAD,and protein levels of Nrf2/HO-1 signal pathway were detected.Results Compared with sham operation group,the neurological deficit score,Evans blue exudation,appptotic rate of hippocampal neurons,serum COX-2,IL-6,iNOS levels,brain tissue reactive oxygen species(ROS)and malondialdehyde level in the model group were significantly increased(P＜0.05),and the expression levels of CAT,GSH-Px,Nrf2 and HO-1 proteins were significantly decreased(P＜0.05).Compared with dandelion extract group,combination of dandelion extract and ML385 significantly increased the neurological deficit score(2.54±0.23 vs 1.43±0.19),Evans blue exudation[(22.15±3.61)ng/mg vs(6.54±1.24)ng/mg],apoptotic rate[(31.97±5.26)％vs(3.51±0.94)％],serum COX-2[(5.82±1.16)ng/ml vs(1.34±0.42)ng/ml],IL-6[(1.47±0.31)ng/ml vs(0.43±0.14)ng/ml]and iNOS levels[(59.91±10.36)U/ml vs(13.94±3.78)U/ml],brain tissue ROS[(4.70±0.45)U/kg vs(1.70± 0.51)U/kg]and MDA levels[(3.72±0.52)nmol/mg vs(1.17±0.34)nmol/mg],and decreased expression levels of CAT[(2.54±0.59)U/mg vs(5.68±1.04)U/mg],GSH-Px[(8.01±0.86)U/mg vs(16.97±3.03)U/mg],Nrf2(0.67±0.13 vs 1.07±0.19)and HO-1(0.55±0.07 vs 0.86± 0.10,P＜0.05).Conclusion Dandelion extract can enhance the antioxidant activity in ICH rats by activating Nrf2/HO-1 signaling pathway,prevent the progression of inflammation and oxida-tive stress,inhibit neuronal apoptosis in hippocampus,repair blood-brain barrier function,and thus improve nerve function.

Background: The impact of para-aortic lymphadenectomy (PALD) on prognosis and quality of life (QoL) for IB2-IIA2 cervical cancer patients remain controversial. And whether intraoperative frozen pathology exam on common iliac lymph nodes could help predict para-aortic lymph node (PALN) metastasis was unanswered with high-level evidence. Methods A multi-center, randomized controlled study is intended to investigate the effect of PALD on the prognosis and QoL in cervical cancer patients and to assess the value of intraoperative frozen pathological evaluation of common iliac nodes metastasis for the prediction of PALN metastasis. After choosing whether to receive intraoperative frozen pathological examination of bilateral common iliac lymph nodes, eligible patients will be randomly assigned (1:1) to receive PALD or not. The primary end point is 2-year progression-free survival (PFS). The secondary end points include 5-year PFS, 2-year overall survival (OS), 5-year OS, adverse events (AEs) caused by PALD, AEs caused by radiotherapy and QoL. A total of 728 patients will be enrolled from 8 hospitals in China within 3-year period and followed up for 5 years.

Objective:To explore the effect and mechanism of diosmetin (Dio) on neuronal ferroptosis in rats with bacterial meningitis (BM).Methods:Male SD rats aged 6-7 weeks of SPF grade were selected for the experiment. The BM model was established by injecting group B hemolytic streptococcus into the cisterna magna of cerebellum. Sixty BM model rats were successfully modeled and divided into model group, low-dose Dio group, medium-dose Dio group, high-dose Dio group and inhibitor group according to the random number table method, with 12 rats in each group. Another 12 weight-matched rats were taken as the control group.The rats in the low-dose Dio group, medium-dose Dio group, high-dose Dio group and the inhibitor group were intragastrically administered with Dio at 50 mg/kg, 100 mg/kg, 200 mg/kg and 200 mg/kg, respectively. The rats in the control group were intragastrically administered with an equal volume of 0.9 % sodium chloride solution. On the day of intragastric administration, the rats in the inhibitor group were intraperitoneally injected with SIRT1 pathway inhibitor EX527 (10 mg/kg), and the rats in the other groups were injected with an equal volume of 0.9% sodium chloride solution. The above interventions were performed once a day for 28 consecutive days. Loeffler neurological score was used to evaluate the neurological impairment in rats. Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in cerebrospinal fluid of rats were detected by ELISA. The number of white blood cells in cerebrospinal fluid was detected by a blood cell analyzer. Glutathione (GSH) was detected by micro-enzyme labeling method, malondialdehyde (MDA) was detected by thiobarbituric acid colorimetric method, reactive oxygen species(ROS) was detected by colorimetry, and Fe 2+ level was detected by ferrozine method. Hematoxylin-eosin staining, Prussian blue staining and TUNEL staining were used to observe the pathological damage, iron accumulation and apoptosis in the hippocampus, respectively.Western blot was applied to measure the expression of transferrin (Tf), proliferating cell nuclear antigen (PCNA), Bcl-2-associated X protein (Bax), caspase-3 and SIRT1/Nrf2/HO-1/Gpx4 signaling pathway proteins. Graphpad Prism 9.0 was used for data analysis. One-way ANOVA was used for statistical analysis, and SNK- q test was used for further pairwise comparisons. Results:(1) There was a statistically significant difference in neurological function scores among the 6 groups of rats ( F=125.451, P<0.001). The neurological function score of the model group was lower than that of control group, while the neurological function scores of the low-dose Dio group, medium-dose Dio group, and high-dose Dio group were higher than those of the model group (all P<0.05). The neurological function score of the inhibitor group ((2.57±0.26)) was lower than that of high-dose Dio group ((4.34±0.48)) ( P<0.05). (2) There were statistically significant differences in the levels of IL-6, TNF-α and the number of white blood cells in the cerebrospinal fluid of rats among the 6 groups ( F=127.817, 102.413, 180.967, all P<0.001). The levels of IL-6, TNF-α and the number of white blood cells in model group were higher than those of control group(all P<0.05). The levels of IL-6, TNF-α and the number of white blood cells in low-dose Dio group, medium-dose Dio group and high-dose Dio group were lower than those of model group (all P<0.001), and those in inhibitor group were all higher than those in high-dose Dio group(all P<0.001). (3) There were statistically significant differences in iron deposition rate and neuronal apoptosis rate among the 6 groups of rats ( F=90.857, 88.835, both P<0.001). The iron deposition rate ((18.37±3.14)%) and neuronal apoptosis rate ((27.58±2.63)%) in the inhibitor group were higher than those in the high-dose Dio group ((6.35±1.08)%, (14.02±1.87)%) (both P<0.05). (4) The levels of GSH, ROS, MDA, and Fe 2+ in the hippocampus of the 6 groups of rats showed statistically significant differences ( F=54.465, 106.453, 55.969, 105.457, all P<0.001). The GSH content in the inhibitor group ((103.48±8.76) mmol/g) was lower than that in the high-dose Dio group ((133.97±10.54) mmol/g), while the contents of ROS, MDA, Fe 2+ ((225.17±16.32) μmol/mg, (10.73±1.58) μmol/mg, (62.71±5.43) μg/g) were higher than those of the high-dose Dio group ((131.87±11.67) μmol/mg, (4.35±0.87) μmol/mg, (34.86±2.95) μg/g) (all P<0.05). (5)There were statistically significant differences in the protein levels of Tf, PCNA, Bax, caspase-3, SIRT1, Nrf2, HO-1 and Gpx4 in the hippocampus of the 6 groups of rats ( F=120.179, 107.568, 157.265, 98.031, 90.932, 52.283, 59.424, 114.539, all P<0.001). The protein levels of Tf, Bax and caspase-3 in the hippocampus of inhibitor group were higher than those of the high-dose Dio group, while the protein levels of PCNA, SIRT1, Nrf2, HO-1, Gpx4 were lower than those of the high-dose Dio group (all P<0.05). Conclusion:Diosmetin can activate SIRT1/Nrf2/HO-1/Gpx4 signaling pathway, thereby inhibiting neuronal ferroptosis in BM rats.

ObjectiveTo investigate the protective effect of modified Huangqi Guizhi Wuwutang （MHGW） on endoplasmic reticulum stress in the sciatic nerve of diabetes rats based on the pathways of inositol-requiring enzyme 1α （IRE1α） and CCAAT/enhancer-binding protein homologous protein （CHOP）. MethodSixty rats were fed on a high-sugar and high-fat diet for six weeks， followed by intraperitoneal injection of streptozotocin at a dose of 35 mg·kg^-1. Random blood glucose levels were measured three days later and rats with a sustained blood glucose level ≥ 16.7 mmol·L^-1 were included in study （n=48）. The rats were randomly divided into a model group， an α-lipoic acid group （0.026 8 g·kg^-1·d^-1）， a high-dose MHGW group （2.5 g·kg^-1·d^-1）， and a low-dose MHGW group （1.25 g·kg^-1·d^-1）. Another 10 rats were assigned to the normal group. The intervention lasted for 16 weeks. After 16 weeks， the sciatic nerve structure of the rats in each group was observed under light microscopy using Luxol fast blue （LFB） staining. Transmission electron microscopy was used to observe the ultrastructure of the sciatic nerve. Chemiluminescence method was employed to measure the serum reactive oxygen species （ROS） levels. Western blot and real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） were used to evaluate the expression of p-IRE1α protein， IRE1α mRNA， CHOP protein， and CHOP mRNA in the sciatic nerve of the rats. ResultCompared with the normal group， the model group showed elevated serum ROS levels （P<0.01）. In contrast， the serum ROS levels were significantly reduced in the treatment groups compared with those in the model group （P<0.01）. The sciatic nerve of the model group showed pathological changes compared with that in the normal group， while the treatment groups exhibited improvement in sciatic nerve pathology compared with the model group. The protein expression of p-IRE1α and CHOP in the sciatic nerve significantly increased in the model group as compared with that in the normal group （P<0.01）. However， the treatment groups showed a significant decrease in the protein expression of p-IRE1α and CHOP in the sciatic nerve compared with the model group （P<0.05， P<0.01）. Furthermore， compared with the normal group， the model group showed upregulated mRNA expression of IRE1α and CHOP in the sciatic nerve （P<0.01）， while the treatment groups exhibited a significant decrease in the mRNA expression of IRE1α and CHOP compared with the model group （P<0.01）. ConclusionMHGW can alleviate endoplasmic reticulum stress-induced cell apoptosis and improve the structure and function of the sciatic nerve in diabetes rats by inhibiting the expression of IRE1α/CHOP pathway-related proteins and mRNA， thereby preventing and treating peripheral neuropathy in diabetes.

ObjectiveTo investigate the effect of modified Huangqi Guizhi Wuwutang （MHGW） on the protein and mRNA expression of B-cell lymphoma-2-associated X protein （Bax） and cysteinyl aspartate specific proteinase-12 （Caspase-12） related to the apoptosis of sciatic nerve cells in diabetes rats to explore the mechanism of MHGW in the treatment of peripheral neuropathy in diabetes. MethodAnimal experiments were conducted. A diabetes model was induced in sixty male sprague-dawley （SD） rats by feeding on a high-sugar and high-fat diet combined with streptozotocin （STZ） intraperitoneal injection. Rats with random blood glucose levels ≥ 16.7 mmol·L^-1 for three consecutive days were considered to have successfully developed diabetes. Forty-eight rats that successfully developed diabetes were randomly divided into a model group， an α-lipoic acid group （0.026 8 g·kg^-1·d^-1）， a high-dose MHGW group （2.5 g·kg^-1·d^-1）， and a low-dose MHGW group （1.25 g·kg^-1·d^-1）， with 12 rats in each group. Another 10 rats were assigned to the normal group. Body weight and random blood glucose levels of the rats were monitored. At the end of a 16-week intervention period， the sciatic nerve conduction velocity of the rats was measured using the Key point electromyography collection system. The protein and mRNA expression of Bax and Caspase-12 in the sciatic nerve cells was detected by Western blot analysis and real-time quantitative polymerase chain reaction （Real-time PCR）， respectively. ResultCompared with the normal group， the model group showed a significant decrease in body weight （P<0.01） and a significant increase in random blood glucose levels （P<0.01）. After a 16-week intervention， compared with the model group， the high-dose MHGW group exhibited a significant increase in body weight （P<0.05）， while there were no statistically significant differences in body weight changes among the other treatment groups. Random blood glucose levels significantly decreased in all treatment groups （P<0.01）. After 16 weeks of intervention， compared with the normal group， the model group had significantly reduced motor and sensory nerve conduction velocities （P<0.01）. Compared with the model group， all treatment groups showed significant increases in motor and sensory nerve conduction velocities （P<0.05， P<0.01）. The expression of Bax and Caspase-12 proteins in the sciatic nerve cells was significantly elevated in the model group compared with that in the normal group （P<0.01）. In contrast， all treatment groups showed significant reductions in the expression of Bax and Caspase-12 proteins in the sciatic nerve cells as compared with that in the model group （P<0.01）. The expression of Bax and Caspase-12 mRNA in the sciatic nerve cells significantly increased in the model group compared with that in the normal group （P<0.01）. Compared with the model group， the α-lipoic acid group and the high-dose MHGW group showed significant reductions in the expression of Bax mRNA in the sciatic nerve cells （P<0.05， P<0.01）， while the low-dose MHGW group showed a decreasing trend in the expression of Bax mRNA. The expression of Caspase-12 mRNA in the sciatic nerve cells significantly decreased in all treatment groups （P<0.01）. ConclusionMHGW may improve and repair sciatic nerve damage in diabetes rats by inhibiting sciatic nerve cell apoptosis.

Background: In China, secondary cytoreductive surgery (SCR) has been widely used in ovarian cancer (OC) over the past two decades. Although Gynecologic Oncology Group-0213 trial did not show its overall survival benefit in first relapsed patients, the questions on patient selection and effect of subsequent targeting therapy are still open. The preliminary data from our pre-SOC1 phase II study showed that selected patients with second relapse who never received SCR at recurrence may still benefit from surgery. Moreover, poly(ADP-ribose) polymerase inhibitors (PARPi) maintenance now has been a standard care for platinum sensitive relapsed OC. To our knowledge, no published or ongoing trial is trying to answer the question if patient can benefit from a potentially complete resection combined with PARPi maintenance in OC patients with secondary recurrence. Methods SOC-3 is a multi-center, open, randomized, controlled, phase II trial of SCR followed by chemotherapy and niraparib maintenance vs chemotherapy and niraparib maintenance in patients with platinum-sensitive second relapsed OC who never received SCR at recurrence. To guarantee surgical quality, if the sites had no experience of participating in any OC-related surgical trials, the number of recurrent lesions evaluated by central-reviewed positron emission tomography–computed tomography image shouldn't be more than 3. Eligible patients are randomly assigned in a 1:1 ratio to receive either SCR followed by 6 cyclesof platinum-based chemotherapy and niraparib maintenance or 6 cycles of platinum-based chemotherapy and niraparib maintenance alone. Patients who undergo at least 4 cycles of chemotherapy and must be, in the opinion of the investigator, without disease progression, will be assigned niraparib maintenance. Major inclusion criteria are secondary relapsed OC with a platinum-free interval of no less than 6 months and a possibly complete resection. Major exclusion criteria are borderline tumors and non-epithelial ovarian malignancies, received debulking surgery at recurrence and impossible to complete resection. The sample size is 96 patients. Primary endpoint is 12-month non-progression rate.

Background: Two randomized phase III trials (EORTC55971 and CHORUS) showed similar progression-free and overall survival in primary or interval debulking surgery in ovarian cancer, however both studies had limitations with lower rate of complete resection and lack of surgical qualifications for participating centers. There is no consensus on whether neoadjuvant chemotherapy followed by interval debulking surgery (NACT-IDS) could be a preferred approach in the management of advanced epithelial ovarian cancer (EOC) in the clinical practice. Methods The Asian SUNNY study is an open-label, multicenter, randomized controlled, phase III trial to compare the effect of primary debulking surgery (PDS) to NACT-IDS in stages IIIC and IV EOC, fallopian tube cancer (FTC) or primary peritoneal carcinoma (PPC).The hypothesis is that PDS enhances the survivorship when compared with NACT-IDS in advanced ovarian cancer. The primary objective is to clarify the role of PDS and NACT-IDS in the treatment of advanced ovarian cancer. Surgical quality assures include at least 50% of no gross residual (NGR) in PDS group in all centers and participating centers should be national cancer centers or designed ovarian cancer section or those with the experience participating surgical trials of ovarian cancer. Any participating center should be monitored evaluating the proportions of NGR by a training set. The aim of the surgery in both arms is maximal cytoreduction. Tumor burden of the disease is evaluated by diagnostic laparoscopy or positron emission tomography/computed tomography scan. Patients assigned to PDS group will undergo upfront maximal cytoreductive surgery within 3 weeks after biopsy, followed by 6 cycles of standard adjuvant chemotherapy. Patients assigned to NACT group will undergo 3 cycles of NACT-IDS, and subsequently 3 cycles of adjuvant chemotherapy. The maximal time interval between IDS and the initiation of adjuvant chemotherapy is 8 weeks. Major inclusion criteria are pathologic confirmed stage IIIC and IV EOC, FTC or PPC; ECOG performance status of 0 to 2; ASA score of 1 to 2. Major exclusion criteria are non-epithelial tumors as well as borderline tumors; low-grade carcinoma; mucinous ovarian cancer. The sample size is 456 subjects. Primary endpoint is overall survival.

OBJECTIVE: To investigate the inhibitory effect of genistein on activation of hepatic stellate cells (HSCs) and the role of the autophagy pathway regulated by PPAR-γ in mediating this effect. METHODS: Cultured HSC-T6 cells were exposed to different concentrations of genistein for 48 h, and HSC activation was verified by detecting the expressions of -SMA and 1(I) collagen; autophagy activation in the cells was determined by detecting the expressions of LC3-II and p62 using Western blotting. The autophagy inhibitor 3-MA was used to confirm the role of autophagy in genistein-induced inhibition of HSC activation. A PPAR-γ inhibitor was used to explore the role of PPAR-γ in activating autophagy in the HSCs. RESULTS: Genistein at concentrations of 5 and 50 μmol/L significantly inhibited the expressions of -SMA and 1(I) collagen ( < 0.05), markedly upregulated the expressions of PPAR-γ and the autophagy-related protein LC3-II ( < 0.05) and significantly down-regulated the expression of the ubiqutin-binding protein p62 ( < 0.05) in HSC-T6 cells. The cells pretreated with 3-MA prior to genistein treatment showed significantly increased protein expressions of -SMA and 1(I) collagen compared with the cells treated with genistein only ( < 0.05). Treatment with the PPAR-γ inhibitor obviously lowered the expression of LC3-II and enhanced the expression p62 in genistein-treated HSC-T6 cells, suggesting the activation of the autophagy pathway. CONCLUSIONS PPAR-γ- regulated autophagy plays an important role in mediating genistein-induced inhibition of HSC activation .
Anticarcinogenic Agents ; pharmacology ; Autophagy ; Collagen Type I ; Genistein ; pharmacology ; Hepatic Stellate Cells ; Humans ; PPAR gamma ; physiology