OBJECTIVE To investigate the effect of phosphotyrosine interaction domain containing 1 (PID1, NYGGF4) onpromotion of IR and HCC, and explore its underlying mechanisms. METHODS Lentivirus were used to mediate the knockdown of PID1 in HFD induced IR mouse model as well as ob/ob mice. Intraperitoneal glucose and insulin tolerance were performed 4 weeks after lentivirus injection. Hydrodynamics-based transfection was applied to induce the liver specific overexpression of PID1. Flow cytometry was exerted to detect the proportion and function of immune cells.qRT-PCR and Western blot were used to detect the expression of downstream pathways of PID1. Liquid chromatography-mass spectrometry (LC-MS) and co-immunoprecipitation (Co-IP) were conducted to identify proteins interacting with PID1.Chromatin immunoprecipitation(ChIP)was operated to measure the modification of H3K4me3 of PID1 promoter.RESULTS PID1 restriction improved insulin resistance,hyperglycemia and fatty liver. Conversely, hepatic knockdown of PID1 attenuated liver xenografted tumor growth. Moreover,PID1 liver-specific protooncogenes via hydrodynamics-based transfection established a primary hepatocellular carcinoma mouse model,induced an immunosuppressive environment,with the reduction of CD3+,CD4+,CD8+T cells,retarded maturation of dendritic cells(DCs),pronounced differentiation of regulatory T cells(Tregs),and recruitment of MDSC.In addition,PID1 overexpression activated prolifer-ation related genes, promoted anti-inflammatory genes, suppressed pro-inflammatory genes, induced glycolysis and lipid metabolism genes to facilitate tumorigenesis in liver. Importantly, PID1 exerted its tumor-promoting function through binding to epidermal growth factor receptor(EGFR)and activation of downstream KRAS/ERK pathway.As such,PID1 exist trimethylation of histone H3 at lysine 4(H3K4me3) modification and IR up-regulated the expression of PID1 by activation the H3K4me3 modification. CONCLUSION PID1 is a new gene that exerts both liver cancer-promoting and insulin resistance inducing function.IR accelerates liver cancer development and progression partially dependent on the activation of PID1.

To investigate the cardioprotective effect of formononetin (FMN) on no-reflow (NR) after myocardial ischemia-reperfusion and its molecular mechanism based on integrated pharmacology and experimental verification, firstly, human breast cancer MCF-7 cells and myocardial NR rats were used to confirm the estrogenic activity and the effect of alleviating NR of FMN, respectively. Male SD rats were divided into Sham, NR, FMN (20 mg·kg^-1) and sodium nitroprusside (SNP, 5.0 mg·kg^-1) groups, which were administered once a day for one week, the experiment was approved by the Ethics Committee of Tianjin University of Traditional Chinese Medicine (TCM-LAEC2019095). The pharmacological analysis and in vivo study of NR rats were integrated to reveal the mechanism of FMN improving NR. The results showed that FMN had estrogenic effect and reduced NR by improving cardiac structure and function, reducing NR, ischemic myocardial area and pathological injury of cardiomyocytes. Integrated pharmacology predicts that the mechanism of FMN improving NR is mainly related to phosphatidyinositol-3-kinase-protein kinase B (PI3K-Akt) signal pathway. Phytoestrogens play a role in cardiovascular protection mainly by activating G protein-coupled estrogen receptor (GPER). GPER is also an important regulator in the upstream of PI3K-Akt signaling pathway. This study found that FMN can significantly activate GPER, p-PI3K, p-Akt and phospho-endothelial nitric oxide synthase (p-eNOS). It has good binding ability with GPER and eNOS protein. In this study, through the integration of pharmacology and experimental evaluation, it is revealed that FMN activates PI3K/Akt/eNOS signal pathway by activating GPER, thus significantly improving NR.

Chronic intermitted hypoxia including sleep breathing disorder leads to brain injury. This study explores the potential therapeutic effects of grape seed proanthocyanidin as a neuroprotective agent. A rat model of chronic intermittent hypoxia was employed, and the animals were given low or high doses of grape seed proanthocyanidin. The ultrastructure changes in the brain, the biochemical components, and the animal behavior were examined. The results showed that with hypoxia exposure, neuronal mitochondria exhibited injuries at ultrastructural level, with increased malondialdehyde (MDA) content and reduced superoxide dismutase (SOD) activity. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining revealed increased cell apoptosis in hippocampus. In Morris water maze the animals showed decreased learning abilities, when compared to normal control. The administration of grape seed proanthocyanidin treatment reversed all these observed changes, and improved the learning behavior. We concluded that grape seed proanthocyanidin could alleviate the brain injury caused by hypoxia from sleep breathing disorder.

Objective To explore the change of apoptosis factor Caspase-3 and Bcl-2 in the injured segment of rat with spinal cord injury after inhibiting lentivirus expression of inflammation factor TNF-α. To study the relationship between Caspase-3, Bcl-2, Bax and TNF-α in spinal cord injury. Mthods Spinal cord contusion model was prepared by Allen method. The relation between tumor necrosis factor alpha and Bcl-2, was predicted by the method of GeneMANIA bioinformatics. The RNA which was packaged by lentivirus constructed the RNA interference model of tumour necrosis factor alpha. After interference of tumor necrosis factor alpha, we used the method of QRT-PCR to assays the mRNA expression of Caspase-3 and Bcl-2 in spinal cord and detect of the localization of Caspase-3 and Bcl-2 by immunohistochemistry. Statistical analysis with SPSS17.0. Results SD rats had paraplegia and urinate retentaion because of spinal cord injury. The result of QRT-PCR showed that in the seventh day after SCC, the expression of Caspase-3 reduced significantly (P < 0.05) and Bcl-2 did not change significantly (P > 0.05). Immunohistochemistry experiment results showed that Caspase-3 Bcl-2 and Bax immunoreactive cells were observed in the neurons and glial cells of both white matter and gray matter in the spinal cord. The results were the same with QRT-PCR.. Conclusion TNF-α in rats after SCC can effectively regulate the ratio of Bcl-2 and Bax , and then regulate the expression of Caspase-3 , which may affect the function of apoptosis and function recovery after spinal cord injury.

OBJECTIVESTo study the inhibitory effect of KF-950 on human acrosin and sperm acrosome.

METHODSHuman acrosin was extracted and purified with 2% acetic acid, and its residual activity was evaluated by BAEE/ADH assay after treated with different concentrations of KF-950. ABC assay was used to observe the effect of KF-950 on human acrosome with Biotin-PSA as a probe.

RESULTS1. The activity of normal sperm acrosin was (37.65 +/- 4.47) U/L. 2. The residual activity was inversely related to the concentration of KF-950 (r = -0.998), and had a dose-response curve. The result could be described by Y = 7.57-1.895X. 3. With increase of KF-950 concentration and prolongation of action time, the staining rate of acrosome obviously dropped (P < 0.01).

CONCLUSIONSKF-950 directly inhibits acrosin activity and assumely injures sperm acrosome. It might be a new kind of highly effective inhibitor.

Acrosin ; antagonists & inhibitors ; Acrosome ; metabolism ; Humans ; Male ; Spermatozoa ; drug effects ; physiology

To characterize the genomic sequence and arrangement of WU polyomavirus (WU virus) identified in clinical specimens collected from children with acute respiratory infections in Beijing, China, the sequences of capsid proteins VP1, VP2, and the large tumor antigen (LTAg), as well as the 5'-terminal sequence of WU virus, were amplified from the clinical specimen with ID number of BJF5276 which was determined as WU virus positive by PCR amplification. The PCR amplicons were sequenced, and genomic sequence analysis was performed by using the software DNAStar. In addition, VP2 coding-region sequences were amplified from other 21 clinical specimens identified as WU virus positive to investigate the gene diversity of WU virus. The genomic sequence of WU virus BJF5276 with accession number of HQ218321 in GenBank was 5,229 base pairs in length with 3 major coding domain sequences (CDS) sited on one strand coding for capsid proteins VP2, VP3 and VP1, and two CDS sited on the complementary strand coding for small tumor antigen (STAg) and LTAg; These 22 VP2 CDS sequences including 5 sequences submitted to GenBank were compared with 64 corresponding sequences downloaded from GenBank by MegAlign of DNAStar software, indicated that these sequences coming from children in Beijing shared high homology (over 98.8%) with those from GenBank. Phylogenetic analysis of these VP2 CDS by using Neighbor-joining (NJ) analyses with 2,000 bootstraps (Mega 4.0) showed that 20 sequences out of 22 belonged to clade Ia, and other 2 of them belonged to clade III, including 1 clustered in IIIa and 1 in a novel cluster proposed as IIIc. In conclusion, the genomic sequence of WU polyomavirus detected from clinical specimens from children in Beijing is closely related to other WU polyomaviruses in the feature of genomic coding region arrangement. Overall variation of VP2 CDS was very low, and there were different clades circulating in Beijing with a dominant clade Ia, which is different from dominated Ib circulating in other parts of the world reported previously, and a novel clade IIIc was proposed.
Acute Disease ; Child, Preschool ; China ; Female ; Genome, Viral ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny ; Polyomavirus ; classification ; genetics ; isolation & purification ; Respiratory Tract Infections ; virology ; Viral Proteins ; genetics

·Corneal confocal microscopy can be used in the real-time, noninvasive, high-resolution corneal structure at the cellular level, which has been widely used in corneal disease research. This review summarizes recent advances in corneal confocal microscopy in the study of infectious keratitis, dry eye, keratoconus, diabetic peripheral neuropathy, and clinical studies to guide corneal transplantation.

OBJECTIVETo explore the impact of being informed of HIV infection before or after pregnancy on the prevention of mother-to-child transmission (PMTCT) HIV interventions uptake.

METHODSFrom 2005 to 2009, a tatal of 5552 HIV-infected pregnant women and their 5894 pregnancies in Henan, Guangxi, Yunnan and Xinjiang province were investigated using the method of a cohort study. The social-demographic characters (the objects were divided three age groups 15-, 25-, 35-49), the period identified to be HIV positive, the outcome of pregnancy and the PMTCT interventions including uptake of antiretroviral drugs (ARVs) were investigated. Through single-factor and non-conditional logistic regression model, the factors influencing the utilization of PMTCT services were analyzed.

RESULTSOf HIV-infected pregnant women, 84.5% (4979/5894) were under 35 year-old, and 56.0% (3108/5552) of them were Han group and the percentage of peasant or unemployment was 85.1% (4727/5552). 86.8% (4815/5552) of these women had junior high school education or less, and the proportion of women knowing HIV infection before the pregnancy was 31.2% (1836/5894). Of HIV positive pregnant women, 31.7% (1869/5894) chose to terminate the pregnancy artificially, and the percentage was 43.8% (805/1836) among those knowing HIV infection before pregnancy. The proportion of the ARVs uptake among HIV positive maternities who delivered was 80.0% (3046/3808), while the percentage among those knowing HIV positive before pregnancy was 92.3% (883/957), which was much higher than it (75.9% (2163/2851)) among the pregnant women knowing HIV infection just during the pregnancy (χ(2) = 120.39, P < 0.05). The results of multivariate analysis showed that the proportion of ARVs' uptake was high among those HIV positive pregnant women knowing to be HIV-infected before pregnancy (versus knowing to be HIV-infected after the pregnancy, OR = 3.91 (95%CI: 3.03 - 5.05)) and age of 15 to 24 year-old (versus age of 35 - 49 year-old, OR = 0.75 (95%CI: 0.57 - 0.98)).

CONCLUSIONIt will promote the HIV-infected pregnant women to receive the PMTCT intervention services if they know their HIV sero-status before pregnancy.

Acquired Immunodeficiency Syndrome ; prevention & control ; psychology ; transmission ; Adolescent ; Adult ; Female ; HIV Infections ; prevention & control ; transmission ; Humans ; Infectious Disease Transmission, Vertical ; prevention & control ; Maternal Health Services ; utilization ; Middle Aged ; Pregnancy ; Pregnancy Complications, Infectious ; prevention & control ; Prospective Studies ; Young Adult

AIM To analyze the effects of α-mangostin on the proliferation and apoptosis of osteoarthritis (OA) chondrocytes.METHODS Human OA chondrocytes were isolated and then treated with 5,10 or 20 μmol/L α-mangostin.24,48 or 72 h after the treatment,the cell proliferation was measured by MTT assay,and the cell apoptosis was detected by flow cytometry.The expression check on MMP-1,MMP-3,MMP-13,PPARγ,PPARδ,PGC-lα and TNF-α was accomplished by Western blot.The contents of collagen-Ⅱ,PG,IL-1β and IL-6 were tested by ELISA.RESULTS α-Mangostin significantly induced cell proliferation and suppressed cell apoptosis,and it significantly increased the production of collagen-Ⅱ and PG,decreased the expressions of MMP-1,MMP-3 and MMP-13,induced the expressions of PPARγ,PPARδ and PGC-1α,and decreased the expression of TNF-α.Furthermore,α-mangostin significantly inhibited the production of IL-1β and IL-6.CONCLUSION α-Mangostin attenuates the destruction and degradation of cartilago articularis by inducing OA chondrocytes proliferation,inhibiting cell apoptosis and inflammation,and increasing expressions of PPARγand PPARδ.

This project's aim was to determine the reserpine-induced gastric ulcer preventive effect of polysaccharide of Larimichthys crocea swim bladder (PLCSB) in ICR mice. The anti-gastric ulcer effects of polysaccharide of Larimichthys crocea swim bladder was evaluated in mice model using morphological test, serum levels assay, cytokine levels assay, tissue contents analysis, reverse transcription-polymerase chain reaction (RT-PCR) analysis and western bolt assay. High concentration (50 mg/kg dose) of PLCSB reduced IFN-gamma as compared to low concentration (25 mg/kg dose) and control mice. SS and VIP serum levels of PLCSB treated mice were higher than those of control mice, and MOT and SP serum levels were lower than control mice. Gastric ulcer inhibitory index of PLCSB treatment groups mice were much lower than control mice, and the high concentration treated mice were similar to the ranitidine treated mice. The SOD and GSH-Px activities of PLCSB treated mice were higher than control mice, close to normal mice and ranitidine treated mice. PLCSB treated mice also showed the similar contents of NO and MDA to normal group. By RT-PCR and western blot assay, PLCSB significantly induced inflammation in tissues of mice by downregulating NF-kappaB, iNOS, and COX-2, and upregulating IkappaB-alpha . These results suggest that PLCSB showed a good gastric ulcer preventive effect as the gastric ulcer drug of ranitidine. Polysaccharide of Larimichthys crocea swim bladder may be used as a drug material from marine products.
Animals ; Blotting, Western ; Inflammation ; Mice ; Mice, Inbred ICR* ; NF-kappa B ; Ranitidine ; Reserpine* ; Stomach Ulcer* ; Ulcer ; Urinary Bladder*