Objective: To investigate the regulatory effect of Jieduan Niwan Formula (JDNWF) drug-containing serum on AMPK-mediated mitochondrial quality control in D-GalN-induced HepG2 cells. Methods: Twenty male Wistar rats were randomly divided into blank control and JDNWF-containing serum groups, 10 rats per group. The JDNWF-containing serum group was gavaged with JDNWF (21.7 g/kg), whereas the blank control group was gavaged with saline. Blood was collected to prepare JDNWF-containing and blank control serum. Cell viability, mitochondrial damage indicators, and MQC pathway protein expression levels were evaluated to determine the optimal volume fraction of JDNWF. HepG2 cells were divided into control, D-GalN, DMSO, AMPK inhibitor, JDNWF drug-containing serum, and JDNWF drug-containing serum plus AMPK inhibitor groups, and corresponding drug interventions were administered to each group. Cells were collected after the interventions, and the CCK-8 assay was used to measure cell viability, the 2′-7′-dichlorodihydrofluorescein diacetate fluorescent probe was used to detect reactive oxygen species (ROS) levels, JC-1 was used to detect mitochondrial membrane potential, thiobarbituric acid was used to measure malondialdehyde (MDA) levels, WST-8 was used to measure superoxide dismutase (SOD) activity, and western blotting was used to detect the expression levels of mitochondrial quality control-related proteins, including p-AMPK, AMPK, PGC-1α, NRF1, TFAM, MFN2, and DRP1. Results: 5% JDNWF drug-containing serum most significantly restored cell viability, mitochondrial damage markers, and MQC pathway protein expression in the model group. Therefore, it was chosen for intervention in subsequent experiments. Compared to the control group, the cell viability of the D-GalN, DMSO, and AMPK inhibitor groups was significantly reduced (P<0.01). In contrast, the heterogeneity of mitochondrial membrane potential, ROS, and MDA levels was significantly increased (P<0.01), and SOD activity was significantly decreased (P<0.01). The p-AMPK, PGC-1α, NRF1, TFAM, MFN2, and DRP1 protein expression levels were significantly decreased (P<0.01). After JDNWF drug-containing serum intervention, compared to the DMSO group, cell viability significantly increased (P<0.01), mitochondrial membrane potential heterogeneity, ROS, and MDA levels significantly decreased (P<0.01), SOD activity significantly increased (P<0.01), and p-AMPK, PGC-1α, NRF1, TFAM, and MFN2 protein expression levels significantly increased (P<0.01), whereas DRP1 protein expression significantly decreased (P<0.01). Compared to the JDNWF drug-containing serum group, the cell viability in the JDNWF plus AMPK inhibitor group significantly decreased (P<0.01), mitochondrial membrane potential heterogeneity and ROS levels significantly increased (P<0.01), MDA levels significantly increased (P<0.05), SOD activity significantly decreased (P<0.05), p-AMPK, PGC-1α, NRF1, and TFAM protein expression levels significantly decreased (P<0.01), MFN2 protein expression significantly decreased (P<0.05), and DRP1 protein expression significantly increased (P<0.01). Conclusion JDNWF drug-containing serum may restore mitochondrial function and improve D-GalN-induced HepG2 cell injury by regulating AMPK-mediated mitochondrial quality control.

BackgroundMetabolic syndrome （MetS） is highly prevalent in patients with mental disorders， including elevated diastolic or systolic blood pressure， elevated fasting glucose， hypercholesterolemia， abdominal obesity and so on. As an important component of MetS， the relationship between hypercholesterolemia and mental disorder has been extensively reported， whereas few genome-wide association studies （GWAS） have been conducted to identify the causal role of mental disorders in hypercholesterolemia. ObjectiveTo explore the potential causal relationship between mental disorders and hypercholesterolemia by two-sample Mendelian randomization （MR） method. MethodsSummary data from GWAS were analyzed. Single nucleotide polymorphisms （SNPs） strongly associated with mental disorders were chosen as instrumental variables， and hypercholesterolemia was used as outcome variable. MR analysis utilized inverse-variance weighted （IVW）， MR-Egger regression and weighted median estimation （WME） as the primary analytical tool， and supplemented by simple mode （SM） and weighted mode （WM）. The causal relationship between mental disorders and the risk of hypercholesterolemia was illustrated in terms of odds ratio （OR）. ResultsA total of 36 SNPs associated with mental disorders were identified as instrumental variables. The primary findings from IVW revealed existence of a causal relationship between mental disorders and hypercholesterolemia （IVW： OR=1.067， 95% CI： 1.026~1.109， P=0.001）. Findings from the additional methods （MR-Egger regression， WME， SM， WM） were basically consistent with those reported in IVW method. Further verification indicated that the causal relationship between mental disorders and the risk of hypercholesterolemia was not affected by genetic polymorphism （P>0.05）. The absence of heterogeneity was confirmed through Cochran's Q test and MR-Egger regression （P>0.05）. Furthermore， no causal association in the reverse direction was found （P>0.05）. ConclusionThere is a causal relationship between mental disorders and hypercholesterolemia， and patients with mental disorders may have an increased probability of suffering from hypercholesterolemia.

Objective:To investigate the status of hypoglycemia fear, diabetes distress, self-regulatory fatigue and self-management in type 2 diabetes patients. The chain mediating effects of self-regulated fatigue and psychological distress on hypoglycemic fear and self-management were investigated.Methods:The Hypoglycemia Fear Survey-Worry Subscale (CHFSII-WS), Diabetes Distress Scale (DDS), Self-Regulatory Fatigue Scale (SRF-S) and Summary of Diabetes Self-Care Activities (SDSCA) were used to investigate the type 2 diabetes patients from the department of Endocrinology of the First Affiliated Hospital in Jinzhou Medical University. And constructed the structural equation model.Results:The scores of fear of hypoglycemia in type 2 diabetes patients were positively correlated with psychological pain and self-regulating fatigue ( r=0.739, 0.625, P<0.05), but negatively correlated with self-management level ( r=-0.602, P<0.05). The psychological pain score was positively correlated with the self-regulating fatigue score ( r=0.669, P<0.05) and negatively correlated with the self-management level score ( r=-0.609, P<0.05). The score of self-regulation fatigue was negatively correlated with the score of self-management ( r=-0.596, P<0.05). Pathway analysis showed that hypoglycemia fear could directly affect self-management behavior, indirectly predict self-management level through self-regulation fatigue and psychological pain respectively, and negatively affect self-management behavior through chain mediation of self-regulation fatigue and psychological pain ( χ2/ df=3.079, GFI=0.920, CFI=0.961, NFI=0.943, IFI=0.961, RMSEA=0.078). Conclusions:The Self-regulated fatigue and psychological distress acts as the chain mediators of hypoglycemic fear and self-management in patients with type 2 diabetes.

Objective:To explore the clinical application value of each sequence by analyzing the characteristics of labyrinthine signal on MRI in patients with unilateral sudden deafness.Methods:Totally 52 patients of unilateral sudden deafness with inner ear MRI were analyzed retrospectively at Beijing Tongren Hospital, Capital Medical University from January 2016 to July 2019, all of which could find abnormalities in the labyrinth, including 17 cases of plain scan and 35 cases of enhanced scan, with sequences including plain T 1WI, enhanced T 1WI, plain and enhanced delayed 3D fluid attenuation inversion recovery (3D-FLAIR). The affected labyrinthine signal characteristics of each sequence were analyzed and the involvement sites were judged. The ability of each sequence to show labyrinthine abnormal signal was evaluated and scored. The Friedman test and Wilcoxon signed rank sum test were used to compare the subjective scores of the ability to show labyrinthine high signal in different sequences in plain and enhanced patients, respectively. Fisher′s exact probability method was used to analyze the relationship between the affected sites and the recovery of hearing, tinnitus and vertigo symptoms. Results:Fifty-two patients (100%, 52/52) showed labyrinthine high signal on T 1WI, 8 (15.4%, 8/52) showed higher signal and 3 (5.8%, 3/52) showed low signal on T 2WI. Thirty-five (100%, 35/35) showed high signal on enhanced T 1WI, among which 27 had enhancement (77.1%, 27/35). Fifty-two (100%, 52/52) showed significant high signal of the affected labyrinth on 3D-FLAIR (17 plain scan, 35 enhanced scan). The scores were 2 (2, 2), 3 (2, 3), 3 (3, 4) and 4 (4, 4) of T 1WI, enhanced T 1WI, plain and enhanced 3D-FLAIR respectively. The overall difference in subjective scores of plain T 1WI, enhanced T 1WI and enhanced 3D-FLAIR in enhanced patients was statistically significant (χ2=64.528, P<0.001), and the comparison between the two was statistically different (all corrected P<0.05). The plain 3D-FLAIR score was higher than the plain T 1WI in patients with a statistically significant difference ( Z=-3.729, P<0.001). Twenty-seven cases (51.9%, 27/52) exhibited high signal at the ampulla of semicircular canals, with a statistically significant difference in the distribution of hearing recovery or not ( P=0.001). Conclusions:Both T 1WI and 3D-FLAIR sequences can effectively identify the labyrinthine high signal, but the latter was better than the former of its ability to display, especially delayed enhanced 3D-FLAIR. The high signal at the ampulla of semicircular canals was a characteristic predictor of non-recovery of hearing.

OBJECTIVE To summarize surgical treatment effects and to discuss surgical strategy for Small acoustic neuroma.METHODS The clinical data of 26 patients with small acoustic neuroma treated with surgery in our department were analyzed retrospectively.25 patients with preoperative hearing grading of C and D received the labyrinthine approach acoustic neuroma resection,and 1 patient with preoperative hearing grading of B received middle fossa approach acoustic neuroma resection.The preoperative and postoperative hearing level,incidence of tinnitus,balance disturbance and weakness of facial muscles were analyzed retrospectively.The control and recurrence of the tumor,the incidence of complications were recorded.RESULTS Total resection was achieved in 25 cases (96％),and in 1 cases achieved sub total resection due to the fact that the tumor was too adhesive to the facial nerve The follow-up time ranged from 6 months to 7 years with 7 cases lost contact.No deaths nor intracranial infection were encountered.Cerebrospinal fluid rhinorrhea was found in 1 cases after the operation and eustachian tube blockage was performed.All patients(100％) had their facial nerve reserved and 16 patients (88％) had their cochlea nerve reserved.17patients(89.5％) had intact facial nerve function and 2 patients suffered from level Ⅲ facial nerve paralysis (House-Brackmann grade) post operatively.6 patients suffered from vertigo pre-operatively and 5 of them reported disappearance of vertigo after the surgery.Of the 13 patients who did not have vertigo before the surgery,1 patient complained sense of dizziness and unsteady walking post-operatively.5 patients (33.3％) reported disappeared or alleviated tinnitus,8 patients (53％) had the same degree of tinnitus and 2 patients reported worsened tinnitus post-operatively.For patients who did not have tinnitus before the surgery,no tinnitus was reported post-operatively.CONCLUSION treatment strategy for small acoustic neuroma requires personalized setting based on the age of the patient,the combined systemic disease,the severity of the symptoms and the growth rate of the tumor.For patients without practical hearing,the tumor could be removed with labyrinthine approach,and the integrity of the cochlear nerve should be preserved as far as possible,for further reconstruction of hearing ability and cochlear implantation.

Objective To evaluate the feasibility,safety and effectiveness of modified associating liver partition and portal vein ligation for staged hepatectomy (ALPPS).Methods The published literatures associated with modified ALPPS were pooled from Embase,Pubmed,Medline,Google Scholar databases.The studies were included or excluded depends on our predetermined criteria.We selected data and performd descriptive analysis from the included studies.Results Five articles were included and reviewed.A total of 62 patients underwent five modified procedures,including monosegment ALPPS (m-ALPPS),anterior approach ALPPS,partial-ALPPS,radiofrequency-assisted liver partition with portal vein ligation (RALPP) and associating liver tourniquet and portal ligation for staged hepatectomy (ALTPS).There were 50 (80.6％) patients diagnosed liver metastatic colorectal cancer.The average operation interval of modified ALPPS was between 8 ～ 22 days and growth rate of future liver remnant (FLR) ranged from 48.7％ to 62.3％,the feasibility to perform ALPPS stage 2 was 98.4％.The incidence of severe postoperative complications were between 11.8％ ～33.3％.The 90-day mortality for monosegment ALPPS,partial-ALPPS and RALPP was 0,while the figure was 8.3％ in ALTPS.The in-hospital morbidities were 5.9％ and 8.3％ for anterior approach ALPPS and ALTPS,respectively,which were 0 in the other three modified groups.Clinical response evaluation,including R0 resection rate,overall survival rate,disease-free and recurrence rates were merely presented 83.3％,80％,50％,50％ in m-ALPPS group,while 100％,100％,95％,5％ in modified ALTPS group.Conclusion Modified ALPPS with improved safety is feasible in clinical practice.However,the effectiveness still needs further studies.

Background and purpose:Long non-coding RNA (lncRNA) could be an important player in cancer biology. Recent studies showed that lncRNA PCGEM1 might be important in the regulation of androgen recep-tor (AR) signaling pathway. We tried to observe the expressions of lncRNA PCGEM1 and AR in prostate cancer, and investigate their role and signiifcance in prostate cancer genesis and progress.Methods:The expression of lncRNA PCGEM1 was observed in prostate cancer by lfuorescencein situ hybridization (FISH) technique. Then detection of AR was performed by immunolfuorescence histochemistry methods. Their co-effective role was checked by RNA pull-down technique.Results:Compared with the AR-independent cell line such as PC3 or DU145, AR-dependent cell line such as LNCaP showed much higher expression of lncRNA PCGEM1 (P<0.01). PCGEM1 and AR could be co-localized in most of these prostate cancer samples, especially in the metastasis samples. Moreover, androgen deprivation promoted the translocation of PCGEM1 into nucleus. RNA pull-down results also proved the co-effective role of PCGEM1 and AR.Conclusion:This study showed that lncRNA PCGEM1 was highly expressed in metastatic prostate cancer. It was related to the progress and malignant behavior of the prostate cancer. Its co-localization with AR may play an important role in prostate cancer genesis and progress.

OBJECTIVETo construct a recombinant lentiviral vector that co-express green fluorescent protein (GFP) and FoxM1 shRNA and establish a prostate cancer cell line with stable FoxM1 down-regulation.

METHODSThree interfering sequences targeting FoxM1 were designed and inserted into the lentiviral vector pHBLV-U6-ZsGreen-Puro. After identification by DNA sequencing, the lentiviral vectors carrying Foxm1 shRNA were packaged in 293 cells. The lentiviral particles were collected to infect human prostate cancer DU-145 cells, and the transfection efficiency was observed under fluorescence microscope; the interference efficiency was assessed using real-time PCR. DU-145 cells with stable FoxM1 down-regulation were screened with puromycin, and the expression level of FoxM1 was detected by Western blotting and the cell growth was observed using MTT assay. The stably transfected cells were examined for cell apoptosis and cell clone formation capacity with flow cytometry and colony formation assay.

RESULTSDNA sequencing demonstrated successful construction of the 3 FoxM1 shRNA lentivirus vectors. Real-time PCR showed a high interference efficiency of FoxM1 shRNA1 vector, which resulted in obvious down-regulation of FoxM1 in DU-145 cells. Western blotting showed that the expression of FoxM1 protein was decreased in FoxM1 shRNA1 lentivirus-transfected cells, which displayed a suppressed cell proliferation, increased apoptosis rate, and attenuated clonogenic ability.

CONCLUSIONWe have successfully established a prostate cancer cell model with stable FoxM1 down-regulation, which shows lowered proliferative and clonogenic activities with increased cell apoptosis.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Forkhead Box Protein M1 ; Forkhead Transcription Factors ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Lentivirus ; Male ; Prostatic Neoplasms ; genetics ; RNA, Small Interfering ; genetics ; Real-Time Polymerase Chain Reaction ; Transfection

Objective To construct a recombinant lentiviral vector that co-express green fluorescent protein (GFP) and FoxM1 shRNA and establish a prostate cancer cell line with stable FoxM1 down-regulation. Methods Three interfering sequences targeting FoxM1 were designed and inserted into the lentiviral vector pHBLV-U6-ZsGreen-Puro. After identification by DNA sequencing, the lentiviral vectors carrying Foxm1 shRNA were packaged in 293 cells. The lentiviral particles were collected to infect human prostate cancer DU-145 cells, and the transfection efficiency was observed under fluorescence microscope; the interference efficiency was assessed using real-time PCR. DU-145 cells with stable FoxM1 down-regulation were screened with puromycin, and the expression level of FoxM1 was detected by Western blotting and the cell growth was observed using MTT assay. The stably transfected cells were examined for cell apoptosis and cell clone formation capacity with flow cytometry and colony formation assay. Results DNA sequencing demonstrated successful construction of the 3 FoxM1 shRNA lentivirus vectors. Real-time PCR showed a high interference efficiency of FoxM1 shRNA1 vector, which resulted in obvious down-regulation of FoxM1 in DU-145 cells. Western blotting showed that the expression of FoxM1 protein was decreased in FoxM1 shRNA1 lentivirus-transfected cells, which displayed a suppressed cell proliferation, increased apoptosis rate, and attenuated clonogenic ability. Conclusion We have successfully established a prostate cancer cell model with stable FoxM1 down-regulation, which shows lowered proliferative and clonogenic activities with increased cell apoptosis.

Objective To construct a recombinant lentiviral vector that co-express green fluorescent protein (GFP) and FoxM1 shRNA and establish a prostate cancer cell line with stable FoxM1 down-regulation. Methods Three interfering sequences targeting FoxM1 were designed and inserted into the lentiviral vector pHBLV-U6-ZsGreen-Puro. After identification by DNA sequencing, the lentiviral vectors carrying Foxm1 shRNA were packaged in 293 cells. The lentiviral particles were collected to infect human prostate cancer DU-145 cells, and the transfection efficiency was observed under fluorescence microscope; the interference efficiency was assessed using real-time PCR. DU-145 cells with stable FoxM1 down-regulation were screened with puromycin, and the expression level of FoxM1 was detected by Western blotting and the cell growth was observed using MTT assay. The stably transfected cells were examined for cell apoptosis and cell clone formation capacity with flow cytometry and colony formation assay. Results DNA sequencing demonstrated successful construction of the 3 FoxM1 shRNA lentivirus vectors. Real-time PCR showed a high interference efficiency of FoxM1 shRNA1 vector, which resulted in obvious down-regulation of FoxM1 in DU-145 cells. Western blotting showed that the expression of FoxM1 protein was decreased in FoxM1 shRNA1 lentivirus-transfected cells, which displayed a suppressed cell proliferation, increased apoptosis rate, and attenuated clonogenic ability. Conclusion We have successfully established a prostate cancer cell model with stable FoxM1 down-regulation, which shows lowered proliferative and clonogenic activities with increased cell apoptosis.