Chromodomain-helicase-DNA-binding protein 1 (CHD1) deletion is among the most common mutations in prostate cancer (PCa), but its role remains unclear. In this study, RNA sequencing was conducted in PCa cells after clustered regularly interspaced palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9)-based CHD1 knockout. Gene set enrichment analysis (GSEA) indicated upregulation of hypoxia-related pathways. A subsequent study confirmed that CHD1 deletion significantly upregulated hypoxia-inducible factor 1α (HIF1α) expression. Mechanistic investigation revealed that CHD1 deletion upregulated HIF1α by transcriptionally downregulating prolyl hydroxylase domain protein 2 (PHD2), a prolyl hydroxylase catalyzing the hydroxylation of HIF1α and thus promoting its degradation by the E3 ligase von Hippel-Lindau tumor suppressor (VHL). Functional analysis showed that CHD1 deletion promoted angiogenesis and glycolysis, possibly through HIF1α target genes. Taken together, these findings indicate that CHD1 deletion enhances HIF1α expression through PHD2 downregulation and therefore promotes angiogenesis and metabolic reprogramming in PCa.
Male ; Humans ; Von Hippel-Lindau Tumor Suppressor Protein/metabolism* ; DNA-Binding Proteins/metabolism* ; Prolyl Hydroxylases/metabolism* ; Hypoxia ; Prostatic Neoplasms/pathology* ; Glycolysis ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Cell Line, Tumor ; DNA Helicases/metabolism*

Gut microbiome sequencing studies have great potential to translate microbial analysis outcomes into human health research. Sequencing strategies of 16S amplicon and whole-metagenome shotgun (WMS) are two main methods in microbiome research with respective advantages. However, how sample heterogeneity, sequencers and library preparation protocols affect the sequencing reproducibility of gut microbiome needs further investigation. This study aims to provide a reference for the selection of sequencing technologies by comparing differences in microbial composition from different sampling sites. The results of three widely adopted sequencers showed that the technical repetition correlation (r= 0. 94) was high in WMS method, while the biological repetition correlation (r = 0. 69) was low. Bray-Curtis distance identified that dissimilarity from biological replicates was larger than that of technical replicates (P<0. 001). In addition, dissimilarity and specific taxonomic profiles were observed between 16S and WMS datasets. Our results imply that homogenization is a necessary step before sample DNA extraction. The sequencers contributed less to taxonomic variation than the library preparation protocols. We developed an empirical Bayes approach that " borrowed information" in calculations and analyzed batch effect parameters using standardized data and prior distributions of (non-) parameters, which may improve population comparability between 16S and WMS and provide a basis for further application to fusion analysis of published 16S and microbial datasets.

Objective To understand the prevalence and disease distribution of migrants within the water source area of the Middle Route Project of South-to-North Water Diversion Project and analyze the influencing factors. Methods By multi-stage stratified cluster sampling method, 307 immigrants and 676 immigrants were selected from Danjiangkou reservoir area, Shiyan city, Hubei Province. Results The prevalence of chronic diseases among in-migration was 51.8%. The prevalence rates of the top five chronic diseases were waist and leg pain, hypertension, gastritis, heart disease and diabetes. The prevalence of chronic diseases among out-migration was 65.1%. The prevalence rates of the top five chronic diseases were waist and leg pain, hypertension, gastritis, heart disease and bronchitis. The rate of chronic disease, the rate of two weeks and the rate of two weeks' visit were higher than that of emigration(all P<0.05). The results of multivariate analysis showed that the factors influencing the chronic diseases of immigrants were 60 years old (OR=59.928，P<0.001), unsatisfied family income (OR=0.377, P=0.002) and poor economic status (OR=0.517, P=0.028). The factors influencing the chronic diseases of immigrants were 60 years old (OR=26.190, P<0.001), smoking (OR=1.856,P=0.004), unsatisfactory family income (OR=0.598,P=0.029) and poor economic status (OR=0.460,P<0.001). Conclusions The prevalence of migrants within a high level is at a high level. Actually raising the income of immigrants is conducive to improving the health condition of immigrants.

OBJECTIVETo evaluate the effects of the 25 Gy ⁶⁰Co irradiation on the physiological and biochemical properties and the functions of the platelets during storage.

METHODSA total of 15 bags of platelets were apheresis-collected from 15 healthy donors, and each bag of platelets were divided into 2 parts, then the platelets were divided into the control group (without 25 Gy ⁶⁰Co irradiation) and the irradiated group (with 25 Gy ⁶⁰Co irradiation) groups. The two groups of platelets were kept under the condition of (22 ± 2) °C and shaken. The Platelet count and pH value were detected on the d 1, d 2, d 3, d 4 and d 5. The variables such as R, K values, α angle and maximal amplitude (MA) were measured by thrombelastography on the same days. Hypotonic shock response (HSR), morphological score were devised.

RESULTSThere were no statistically significant difference in Plt counts, mean platelet volume (MPV), platelet distribute width (PDW) and pH between the two groups (P > 0.05), and Plt count decreased on the end of storage. There were no marked changes in HSR level and morphological score between the two groups during storage, and there were no significant difference between the two groups (P > 0.05). In the TEG analysis there were no significant difference of the R, K, α angle and MA values between the two groups (P > 0.05). R value showed upward trend increased along with prolongation of preserved time (P < 0.01), no significant changes in α angle (P > 0.05), K value was slightly higher and MA value was lower in the last day of storage than the days 1-4 (P < 0.01), respectively.

CONCLUSION25 Gy ⁶⁰Co gamma-ray irradiation can not damage the physiological, biochemical properties and the functions of the platelets during storage. In order to ensure the best curative effect, it is suggested that no matter the platelets were irradiated or not, the platelets should be used as soon as possible.

Blood Platelets ; radiation effects ; Blood Preservation ; Gamma Rays ; Humans ; Platelet Count

Objective To investigate the effect of controlled low central venous pressure(CLCVP) on blood loss and prognosis in different types of hepatectomy .Methods Two hundred and fifty seven patients underwent standard hepatectomy ,half liver resec‐tion or irregular partial hepatectomy from January 2011 to December 2012 in the First Affiliated Hospital of Chongqing Medical U‐niversity were retrospectively studied .Patients treated with CLCVP during hepatectomy were attributed to the CLCVP group .CVP of these patients were lowed to below 5 cm H2 O by minimizing fluid infusion and one or both of the following maneuvers :posture adjustment ,nitroglycerin administration .Alpha agonists were used when necessary to maintain the mean arterial pressure MAP at ≥60 mm Hg .Other patients been maintained with normal level of CVP by adjusting fluid administration were included in normal CVP group (NCVP) .Blood loss and transfusion volume ,length of hospital stay of the two groups were compared ,and the effects of different surgery type on CLCVP blood protection were evaluated .Results In the patients underwent standard hepatectomy or half liver resection ,intraoperative blood loss and transfusion were not statistically different between the two groups .While in the pa‐tients underwent irregular partial hepatectomy ,the CLCVP group suffered less blood loss and transfusion(P<0 .05) .Percentage of the patients with less than 200 mL blood loss and no transfusion of concentrated red cell in CLCVP group was higher than that of in NCVP group(P<0 .05) .Differences between the two groups in postoperative hospital stay were with no significance in all the operation types(P>0 .05) .Conclusion The efficiency of CLCVP on blood protection during hepatectomy is influenced by the sur‐gery type ,the blood protection is found to be significant only in irregular partial hepatectomy .No relationship was found between CLCVP and postoperative hospital stay in all types of hepatectomy .

Objective To investigate the role of autophagy and synaptophysin (SYP) in cognitive impairment in de?pressed rats receiving electroconvulsive shock (ECS). Methods Clean and healthy adult male Sprague-Dawley rats were acclimatized to a standard laboratory environment for 7 days. The chronic unpredictable mild stress (CUMS) was used to establish the rat model of depression. Behavior tests were conducted before and after CUMS to evaluate the depression and cognition level of rats. After establishment of the model, 24 rats were randomly divided into ESC group (group E) and depression group (group D) with 12 rats in each group. The rats in group E were administered 80 mg/kg of propofol (10 mg/mL) by intraperitoneal injection, followed by ECS treatment. The rats in group D were administered propofol by intra?peritoneal injection, followed by sham-ECS treatments. The above interventions were conducted daily for 7 consecutive days. After the interventions, rats underwent behavior tests as before. Subsequently, rats were killed and specimens were collected for measurements. Immunohistochemistry was performed to examine autophagy markers such as Beclin 1 and LC3Ⅱand ELISA was used to detect SYP in the hippocampus. Results Group E after ECS significantly increased the percentage of sucrose preference (68.2%±8.7%), rearing times (7.0±1.9), total horizontal distance [(569.5±70.0) cm], es? cape latency [(21.9±5.3)s] and space exploration time [(20.5±3.9)s] compared with group D or group E before ECS. There was no significant difference in these index between groups before ECS or in group E between before and after ECS(P>0.05). Compared with group D, group E had upregulated protein expression levels of Beclin 1 and LC3Ⅱin CA1, CA3, DG as well as the area near the hippocampus and increased SYP contents (P<0.05). Conclusions Cognitive impairment in depression rats following ECS correlates with activated autophagy and increased SYP by ECS.

AIM: To illuminate the molecular targets for schisandrin against cerebrovascular disease based on the combined methods of network pharmacology prediction and experimental verification. METHOD: A protein database was established through constructing the drug-protein network from literature mining data. The protein-protein network was built through an in-depth exploration of the relationships between the proteins. The computational platform was implemented to predict and extract the sensitive sub-network with significant P-values from the protein-protein network. Then the key targets and pathways were identified from the sensitive sub-network. The most related targets and pathways were also confirmed in hydrogen peroxide (H2O2)-induced PC12 cells by Western blotting. RESULTS: Twelve differentially expressed proteins (gene names: NFKB1, RELA, TNFSF10, MAPK1, CHUK, CASP8, PIGS2, MAPK14, CREB1, IFNG, APP, and BCL2) were confirmed as the central nodes of the interaction network (45 nodes, 93 edges). The NF-κB signaling pathway was suggested as the most related pathway of schisandrin for cerebrovascular disease. Furthermore, schisandrin was found to suppress the expression and phosphorylation of IKKα, as well as p50 and p65 induced by H2O2 in PC12 cells by Western blotting. CONCLUSION The computational platform that integrates literature mining data, protein-protein interactions, sensitive sub-network, and pathway results in identification of the NF-κB signaling pathway as the key targets and pathways for schisandrin.
Animals ; Cerebrovascular Disorders ; drug therapy ; genetics ; metabolism ; Cyclooctanes ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Gene Regulatory Networks ; drug effects ; Humans ; Lignans ; pharmacology ; Molecular Targeted Therapy ; PC12 Cells ; Polycyclic Compounds ; pharmacology ; Protein Interaction Maps ; drug effects ; Rats ; Signal Transduction ; drug effects

OBJECTIVETo investigate the molecular mechanisms of icariin (ICA) in regulating the bone formation of osteoblasts and the bone resorption of osteoclasts.

METHODSPrimary osteoblast cell cultures were obtained from newborn rat calvarial. Calcified nodules were stained by alizarin red. The mRNA levels of osterix (OSX), runt-related transcription factor 2 (Runx-2), alkaline phosphatase (ALP), Collagen1, osteoprotegerin (OPG), and receptor activator of nuclear factor-ΚB ligand (RANKL) were analyzed by quantitative real-time RT-PCR, the protein levels of OPG, RANKL, and Collagen1 were examined by Western blotting, and the intracellular Ca(2+) concentration of osteoblasts was measured on a flow cytometer using the Cellquest program.

RESULTSCompared with control group, ICA markedly promoted bone formation by significant up-regulating the gene expressions of OSX, Runx-2,ALP, and Collagen1, the protein expression of Collagen1(all P<0.01), and the Ca(2+) concentration. Furthermore, ICA remarkably inhibited bone resorption by significant up-regulating the mRNA and protein expressions of OPG as well as the OPG/RANKL ratio.

CONCLUSIONSICA could promote bone formation of osteoblasts through inducting the gene expressions of OSX,Runx-2, ALP and Collagen1, and the protein expressions of Collagen1, and by increasing the Ca (2+) concentration. Moreover, ICA could inhibit bone resorption of osteoclasts through regulating OPG/RANKL signal pathway.

Alkaline Phosphatase ; metabolism ; Animals ; Bone Resorption ; Cells, Cultured ; Collagen Type I ; metabolism ; Core Binding Factor Alpha 1 Subunit ; metabolism ; Flavonoids ; pharmacology ; Gene Expression ; Osteoblasts ; drug effects ; Osteoclasts ; drug effects ; Osteogenesis ; drug effects ; Osteoprotegerin ; metabolism ; RANK Ligand ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor Activator of Nuclear Factor-kappa B ; metabolism ; Transcription Factors ; metabolism

OBJECTIVETo investigate the value of susceptibility weighted imaging(SWI)in the diagnosis of hemorrhagic foci early after blast injury and its role in the outcome prediction.

METHODSTotally 30 rabbits with blast-induced cerebral blast injury were used in this study. After routine CT/MRI and SWI scanning,quantified analysis was performed in regions of interest using post-processing technology. After dissecting the brains of the experimental rabbits,the cerebral histopathological changes were observed,and the results were compared with SWI findings.

RESULTSIn these 30 rabbits,22,102,221,and 738 hemorrhagic foci were detected by CT,T1WI,T2WI,and SWI,respectively. The number of cerebral microbleeds detected by SWI was significantly larger than those revealed by conventional T1WI and T2WI(Χ(2)=10.00,P<0.01). Furthermore,the SWI imaging displayed the punctiform(n=315,42.7%),lamellar(n=218,29.5%),slinar(n=205,27.8%)hypointense foci,with clear margin. The number of hemorrhagic foci detected by SWI was positively correlated with survival(r=-0.667,P<0.05).

CONCLUSIONSSWI remarkably increases the detection rate of hemorrhagic foci(particularly microbleeds)in rabbits with cerebral blast injury. The number of cerebral microbleeds and location of foci are closely related with the outcomes and therefore may facilitate clinical managment.

Animals ; Blast Injuries ; complications ; diagnosis ; Brain ; pathology ; Brain Injuries ; diagnosis ; etiology ; Cerebral Hemorrhage ; diagnosis ; etiology ; Female ; Image Enhancement ; Magnetic Resonance Imaging ; methods ; Male ; Prognosis ; Rabbits

OBJECTIVETo compare the effects of 8-prenylnaringenin (PNG) and naringenin (NG) on the activity and apoptosis of osteoclasts cultured in vitro, in order to study physiological activity of 8-prenyl perssad.

METHODOsteoclasts were separated from long-limb bones of newly born rabbits, cultured in alpha-MEM containing 10% FBS, and then added with PNG and NG with the concentration of 1 x 10(-5) mol x L(-1). They were stained with TRAP and determined for enzymatic activity with TRAP after 4 d, and analyzed by toluidine blue staining after 7 d. The apoptotic osteoclasts were analyzed by Annexin V-FITC staining after 2, 4, 8, 12, 24, 36, and 48 hours, to observe their apoptosis. Their total RNAs were extracted, and analyzed for TRAP and Cathepsin K expressions by Real-time RT-PCR.

RESULTCompared with the control group, both of the PNG group and the NG group showed much less osteoclasts (TRAP positive cells), lower TRAP activity and TRAP and Cathepin K (CTSK) expression, and smaller number of bone resorption pits and areas. The PNG group show lower indexes than the NG group. Additionally, the PNG group reached the apoptotic peak of osteoclasts at 12 h after drug administration, whereas the NG group reached after 24 h. And the former had more apoptotic cells than the latter.

CONCLUSION8-PNG is much more active than NG in inhibiting the resorption of osteoclasts and inducing apoptosis of osteoclasts. Their only difference lies in 8-prenyl perssad, which is proved to be able to enhance the anti-bone resorption activity of 8-prenylnarigenin.

Acid Phosphatase ; metabolism ; Animals ; Bone Resorption ; prevention & control ; Cathepsin K ; metabolism ; Cells, Cultured ; Flavanones ; pharmacology ; Osteoclasts ; drug effects ; Rabbits