[Investigation of the effects and mechanisms of berberine on a mouse model of polycystic ovary syndrome: based on intestinal flora analysis].

Objective: To examine the impact of berberine on polycystic ovary syndrome (PCOS) in mice, and to investigate the effects of berberine on the intestinal flora and the intestinal flora on PCOS. Methods: A mouse model of PCOS was established by administering dehydroepiandrosterone in combination with high fat diet, and the mouse model was given a berberine treatment. The study consisted of a blank control group (C group), a PCOS model group (M group) and a berberine treatment group (T group). During the experiment, the mice were closely monitored through timed body weight measurements and estrous cycle monitoring; intraperitoneal glucose tolerance test and insulin tolerance test were done. Upon completion of the pharmacological intervention, the wet weights of liver, ovary and fat deposits of mice were assessed and subjected to HE staining to confirm the success of PCOS modeling and the efficacy of berberine. Additionally, fecal samples were analyzed for intestinal flora through 16S rRNA analysis. Results: The PCOS model was established successfully, berberine alleviated the disturbance of estrous cycle in mice, and significantly alleviated fat accumulation and metabolic abnormalities of glucose in mice. The cross-sectional area of fat pad cells in T group was (2 858±146) µm², which was significantly lower than that in M group [(9 518±347) µm²], and the difference was statistically significant (P<0.001). The blood glucose levels in T group were significantly lower than those in M group (P<0.05). The composition and structure of intestinal flora in mice of M group with PCOS (compared with C group) and in mice of T group after berberine intervention (compared with M group) were significantly altered. However, alpha diversity did not change significantly among three groups (P>0.05). Conclusion: Berberine could alleviate PCOS by intervening in the alterations of gut microbiota.

The mechanism of Epimedium in the treatment of coronary atherosclerotic heart disease based on network pharmacology, molecular docking, and in vitro studies.

OBJECTIVE: There are many challenges related to the treatment of coronary atherosclerotic heart disease (CAD). Studies have confirmed that Epimedium extract inhibits myocardial injury induced by myocardial ischaemia, but the mechanism of action remains unclear. This study aimed at analysed the effective components and mechanisms of Epimedium in treating CAD based on network pharmacology and molecular docking studies and to verify the mechanism in vitro. MATERIALS AND METHODS: The TCMSP and UniProt databases were used to filter for the active components and drug targets of Epimedium. The GeneCards database was used to screen disease targets associated with CAD. The intersection of the drug targets of Epimedium and the disease targets of coronary heart disease was studied to identify the targets of Epimedium in the treatment of CAD. Cytoscape software was used to establish and analyse an activity-target network. The STRING database was used to analyse a protein-protein interaction (PPI) network, and proteins in the PPI network were visualized in the R language. Bioconductor software was used for GO function and KEGG pathway enrichment analyses, and visualization analysis was performed in the R language. PyMOL software was used to verify the molecular docking between selected active components of Epimedium and the targets of CAD, and the potential key effective components of Epimedium in the treatment of coronary heart disease were identified. The involvement of the PI3K/Akt pathway was validated by Western blot analysis. RESULTS: (1) Twenty-three active compounds, including Epimedium glycoside, quercetin, luteolin, and olive resin, were screened out. There were 68 common targets of Epimedium and CAD, including IL-6, ESR1, RELA, FOS, NCOA1, CCND1, EGFR, MAPK8, VEGFA, and CASP8. The potential signaling pathways involved in the treatment of CAD by Epimedium included the human cytomegalovirus infection pathway, the PI3K-Akt signaling pathway, the TNF signaling pathway, and the HIF-1 signaling pathway. (2) Luteolin, quercetin, sitosterol, and anhydroicaritin showed strong binding to targets of CAD based on molecular docking studies. (3) Epimedium extract increased the expression of PI3K, Akt and P-Akt but decreased the expression of IL-6  in vitro. CONCLUSIONS: (1) Icariin, quercetin and luteolin may act on target proteins, including IL-6, ESR1, EGFR, MAPK8, VEGFA and CASP8, to participate in the regulation of the human cytomegalovirus infection pathway, the PI3K-Akt signaling pathway, the TNF signaling pathway and other signaling pathways in order to effectively treat CAD. (2) In vitro studies confirmed that Epimedium extract can treat CAD by upregulating PI3K, Akt and P-Akt protein expression and downregulating IL-6 protein expression in SD rat cardiomyocytes.

Gammaherpesvirus Usurps Host IL-17 Signaling To Support the Establishment of Chronic Infection.

Gammaherpesviruses establish lifelong infection and are associated with a variety of cancers, including B cell lymphomas. These viruses manipulate the B cell differentiation process to establish lifelong infection in memory B cells. Specifically, gammaherpesviruses infect naive B cells and promote entry of both infected and uninfected naive B cells into germinal centers, where the virus usurps rapid proliferation of germinal center B cells to exponentially increase its cellular latent reservoir. In addition to facilitating the establishment of latent infection, germinal center B cells are thought to be the target of viral transformation. In this study, we have uncovered a novel proviral role of host interleukin 17A (IL-17A), a well-established antibacterial and antifungal factor. Loss of IL-17A signaling attenuated the establishment of chronic gammaherpesvirus infection and gammaherpesvirus-driven germinal center response in a route of inoculation-dependent manner. Further, IL-17A treatment directly supported gammaherpesvirus reactivation and de novo lytic infection. This study is the first demonstration of a multifaceted proviral role of IL-17 signaling.IMPORTANCE Gammaherpesviruses establish lifelong infections in a majority of humans and are associated with B cell lymphomas. IL-17A is a host cytokine that plays a well-established role in the clearance of bacterial and fungal infections; however, the role of IL-17A in viral infections is poorly understood. In this study, we show that IL-17A signaling promoted the establishment of chronic gammaherpesvirus infection following the mucosal route of infection, viral lytic replication, and reactivation from latency. Thus, our study unveils a novel proviral role of IL-17A signaling in gammaherpesvirus infection.

Study on diagnostic value of P1NP and ß-CTX in bone metastasis of patients with breast cancer and the correlation between them.

OBJECTIVE: This study aimed to investigate the diagnostic value of the total amino-terminal propeptide of type 1 procollagen (P1NP) and C-terminal telopeptide of ß-I collagen (ß-CTX) in bone metastasis of patients with breast cancer and the correlation between them. PATIENTS AND METHODS: The medical records of 73 patients were retrospectively analyzed. These patients with breast cancer were treated in Oncology, General Surgery, and Orthopedic Departments in The Third People's Hospital of Qingdao from March 2014 to April 2017, including 40 patients with bone metastasis (bone metastasis group) and 33 patients with no bone metastasis (non-bone metastasis group). Other 40 healthy people who received physical examination in the same period were selected as the control group. The expression of P1NP and ß-CTX in plasma were detected by the Enzyme-linked immunosorbent assay, and the correlation between them was analyzed. RESULTS: There were significant differences in P1NP and ß-CTX concentrations among the three groups (p<0.05). The concentrations of P1NP in the control group and the non-bone metastasis group were significantly lower than that in the bone metastasis group (p<0.05); the concentrations of ß-CTX in the control group and the non-bone metastasis group were significantly lower than that in the bone metastasis group (p<0.05). P1NP: AUC=0.852, sensitivity: 72.5%, specificity: 93.9%, CUT OFF=66.44. ß-CTX: AUC=0.883, sensitivity: 85.0%, specificity: 84.8%, CUT OFF=69.8. Joint detection: AUC=0.952, sensitivity: 84.8%, specificity: 99.5%, CUT OFF=99.5. The results of the concentrations of P1NP and ß-CTX in the bone metastasis group detected by the Pearson correlation analysis showed that their concentrations were positively correlated in the bone metastasis group (r=0.764, p<0.05). CONCLUSIONS: P1NP and ß-CTX in plasma have a high diagnostic value for bone metastasis of breast cancer and have important significance in the diagnosis of bone metastasis and disease monitoring.

[Analysis of related factors for primary hepatic carcinoma caused by chronic hepatitis B and hepatitis C].

Objective: To explore the related factors for primary hepatic carcinoma (PHC) caused by chronic hepatitis B (CHB) and hepatitis C (CHC). Methods: According to the principle of cross-sectional study, a cluster random sample method was used, a total of 366 chronic hepatitis patients in hospitals were recruited from three provincial tertiary hospitals in Shanxi, Henan and Jilin between July 2016 and October 2016, respectively. Using a self-designed unified questionnaire, face-to-face interviews was conducted on subjects, including sex, age, alcohol consumption, coffee consumption, green tea consumption, fish consumption, smoking, HBV/HCV diagnosis and treatment, diabetes mellitus, family history of PHC (whether PHC in first-degree relatives), etc. Multivariate unconditional logistic regression were performed to identify the related factors for PHC with CHB and CHC. According to the clinical diagnosis the patients were divided into a chronic hepatitis group (not developing to PHC) and a PHC group. Results: Among 366 cases patients, 287 (78.4%) cases were male, 79 cases were female (21.6%), average age was (52.7±9.3) years. 202 cases were chronic hepatitis group, 164 cases were PHC group. Multivariate unconditional logistics regression analysis indicated that alcohol consumption (odds ratio (OR)=2.11, 95%CI: 1.18-3.75), family history of PHC (OR=5.12, 95%CI: 2.60-10.08) were positively correlated with the development of PHC in chronic b, green tea consumption (OR=0.45, 95%CI: 0.23-0.88), antiviral treatment (OR=0.19, 95%CI: 0.11-0.32) were negatively correlated. Alcohol consumption (OR=3.98, 95%CI: 1.14-13.85) was positively correlated with the development of PHC in chronic c, antiviral treatment (OR=0.14, 95%CI: 0.04-0.50) was negatively correlated. Conclusion: Alcohol consumption, family history of PHC, green tea consumption and antiviral treatment were the related factors for the development of PHC in chronic hepatitis b. Alcohol consumption and antiviral treatment were the related factors for the development of PHC in chronic hepatitis c.

Mechanism of juglone-induced apoptosis of MCF-7 cells by the mitochondrial pathway.

This study investigated the nature and mechanism of juglone-induced apoptosis in the human breast cancer cell line MCF-7. The inhibitory effect of juglone on MCF-7 cell growth was evaluated by the dimethylthiazol tetrazolium assay. Morphological apoptotic changes were characterized using an inverted microscope, Hoechst 33258 fluorescence staining, and Giemsa staining. The rate of cell apoptosis, intracellular levels of reactive oxygen species (ROS), and mitochondrial membrane potential were detected using flow cytometry. Intracellular Ca(2+) concentrations were detected using laser scanning confocal fluorescence microscopy. Expression of the proteins Bcl-2, Bax, and cytochrome C was assessed by western blotting. Caspase-3 activity was quantified using a caspase-3 activity kit. Juglone inhibited the growth of MCF-7 cell line with an IC50 of 11.99 µM. The rates of MCF-7 cell apoptosis at 24 h after exposure to 5, 10, and 20 µM juglone were 9.29, 20.67, and 28.39%, respectively; compared to unexposed cells, juglone-exposed cells exhibited significant elevation in intracellular ROS level, decrease in mitochondrial membrane potential, and increase in intracellular Ca(2+) concentration. Juglone upregulated the expression of Bax, and downregulated the expression of Bcl-2, promoting the release of cytochrome C, thereby upregulating the activity of caspase-3. The results suggest that the mechanism of juglone-induced apoptosis in MCF-7 cells is characterized by elevated ROS levels, reduced Bcl-2 expression, increased Bax expression, decreased mitochondrial membrane potential, increased intracellular Ca(2+) concentration, outer mitochondrial-membrane rupture, cytochrome C release, and caspase-3 activation.

Integrated analysis of gene expression profile and genetic variations associated with ovarian cancer.

OBJECTIVE: Ovarian cancer is the most lethal gynecologic cancer worldwide, since most patients are diagnosed at an advanced stage. To improve the early diagnosis and treatment of ovarian cancer, we performed a integrated analysis of transcription profile and genetic variations to study on the molecular pathogenesis in ovarian cancer. METHODS: mRNA expression profiles of ovarian cancer and normal controls downloaded from ArrayExpress database were applied to identify differentially expressed genes (DEGs). The chromosomal distributions of these DEGs were established using DAVID. Then, DNASeq data from the Cancer Genome Atlas (TCGA) were extracted to analyze gene mutational information including the number of mutations (mut), the number of mutational genes (mutG) and chromosomal distributions of mutations. Statistical method was offered to carrying on correlation analysis of gene mutations and differential expression. RESULTS: A total of 1732 DEGs were identified, and the chromosomal distributions of 97 genes were unknown. These DEGs were most significantly distributed on chromosome 4 with p value = 1.34E-7. Chromosome 1 enriched the most DEGs (11.56%). Statistical algorithm showed that DEGs presented significantly positive correlation with mut (p = 0.000009) and mutG (p = 0.00001). In 48.7% DEGs, gene mutations were found. CONCLUSIONS: We conducted scientific analysis on integration of DEGs in expression profiles and genetic mutations in ovarian cancer, displayed the correlation of differential expression and genetic variations. The result indicated that expression profiles were significantly correlated to genotype.

Isolated sacral metastases as the initial presentation from an endometroid ovarian carcinoma: a case report.

Bone metastases are rarely in ovarian carcainoma. It usually occurrs only when the cancer is advanced or recurrent. A case of endometrioid carcinoma in right ovary with intact capsule is reported. The isolated sacral metastasis was found as the initial presentation, and no distant metastases were reported.

Magnetic graphene-based nanotheranostic agent for dual-modality mapping guided photothermal therapy in regional lymph nodal metastasis of pancreatic cancer.

Although regional lymph nodes (RLN) dissection remains the only way to cure pancreatic cancer metastasis, it is unavoidably associated with sizable trauma, multiple complications, and low surgical resection rates. Thus, exploring a treatment approach for the ablation of drug-resistant pancreatic cancer is always of great concern. Moreover, reoperative and intraoperative mapping of RLN is also important during treatment, because only a few lymph nodes can be detected by the naked eye. In our study, graphene oxides modified with iron oxide nanoparticles (GO-IONP) as a nanotheranostic agent is firstly developed to diagnose and treat RLN metastasis of pancreatic cancer. The approach was designed based on clinical practice, the GO-IONP agent directly injected into the tumor was transported to RLN via lymphatic vessels. Compared to commercial carbon nanoparticles currently used in the clinic operation, the GO-IONP showed powerful ability of dual-modality mapping of regional lymphatic system by magnetic resonance imaging (MRI), as well as dark color of the agent providing valuable information that was instrumental for surgeon in making the preoperative plan before operation and intraoperatively distinguish RLN from surrounding tissue. Under the guidance of dual-modality mapping, we further demonstrated that metastatic lymph nodes including abdominal nodes could be effectively ablated by near-infrared (NIR) irradiation with an incision operation. The lower systematic toxicity of GO-IONP and satisfying safety of photothermal therapy (PTT) to neighbor tissues have also been clearly illustrated in our animal experiments. Using GO-IONP as a nanotheranostic agent presents an approach for mapping and photothermal ablation of RLN, the later may serve as an alternative to lymph node dissection by invasive surgery.

OX40 (CD134) blockade inhibits the co-stimulatory cascade and promotes heart allograft survival.

BACKGROUND: CD134 (OX40) is a member of the tumor necrosis factor receptor superfamily that is expressed as a late event in T-cell activation and contributes to the generation of immunologic memory. CD134 blockade effectively ameliorates inflammation in models of autoimmune disease, but its role in transplantation has been less well studied. METHODS: The authors used an OX40-immunoglobulin (Ig) fusion protein to examine the contribution of this co-stimulatory molecule to the in vitro alloimmune response and studied the ability of CD134 blockade to prevent cardiac allograft rejection in mouse models of heart transplantation using strains representing both major histocompatibility complex (MHC) (BALB/c to CBA/Ca) and minor histocompatibility complex (mHC) (B10.BR to CBA/Ca) antigen mismatches. RESULTS: CD134 upregulation on in vitro alloactivated T cells was delayed compared with CD69 and CD25, and inhibition of T-cell proliferation was critically dependent on the timing of OX40-Ig administration. Heart allograft survival in a fully allogeneic, MHC-mismatched strain combination was not influenced by CD134 blockade alone, but OX40-Ig treatment in the mHC-mismatched model resulted in long-term graft survival (median survival time extended from 14 days to >100 days). Early mononuclear cell infiltration of the graft was similar in both rejecting and long-surviving heart grafts, but OX40-Ig treatment appeared to delay cellular infiltration. CONCLUSIONS: These results show that CD134-CD134L interaction plays an important role in the co-stimulatory cascade and that blockade of this molecular interaction may be of therapeutic value in helping to prevent allograft rejection.

[Experimental circumferential tracheal replacement with rib and costal muscle ring in dog.].

BACKGROUND: To evaluate the feasibility of using vascularized rib and costal muscle ring to repair large circumferential defect of thoracic trachea in dog. METHODS: Eighteen adult mongrel dogs were used in this study. The third rib with adjacent intercostal muscles was engineered to a ring with vascular pedicle , which replaced five or eight rings of trachea. RESULTS: Eleven of eighteen dogs were alive after operation , and the longest one had survived for 14 months. These dogs were active , although a slight stricture at the newly formed trachea was found. Microscopic examination showed the trachea had good blood supply , and the substitution of pleural epithelium by ciliated epithelium was observed. CONCLUSIONS: Circumferential tracheal replacement with vascularized rib and intercostal muscle ring is easily managed and has less complications. The results suggest that this technique might be clinically used in the future.

The complete nucleotide sequence of prune dwarf ilarvirus RNA 3: implications for coat protein activation of genome replication in ilarviruses.

The complete nucleotide sequence of prune dwarf ilarvirus (PDV) RNA 3 has been determined from cloned viral cDNAs. The PDV RNA 3 is 2129 nucleotides and contains two large open reading frames (ORFs) separated by an intergenic region of 72 nucleotides. The 5' proximal ORF (ORF-1) is 882 nucleotides, encoding a gene product which shares homology with putative cell-to-cell movement proteins of related viruses, including tobacco streak virus (TSV) and alfalfa mosaic virus (AIMV). The downstream ORF (ORF-2) is 657 nucleotides and encodes a gene product which shares primary sequence homology and structural features with AIMV coat protein. Furthermore, when expressed in bacteria, this ORF produces a polypeptide which comigrates with authentic PDV coat protein and reacts with PDV coat protein antiserum. Hybridization data suggest that the genomic organization of PDV RNAs 3 and 4 is similar to that of TSV, the only other ilarvirus for which sequence information is published. The 3' untranslated region (UTR) of PDV RNA 3 is similar to that of TSV and AIMV, containing a potential stem-loop structure followed by the sequence AUGC, a structure which may signal binding of coat protein and activation of genome replication. However, a striking feature of the deduced PDV coat protein sequence is the absence of a "zinc-finger" motif thought to function in binding of the coat protein to the 3'-UTR in ilarviruses and AIMV. This result suggests that the zinc-finger motif is not a required aspect of coat protein activation of replication in ilarviruses.