Membrane-Based Preparation Process and Antioxidant and Anti-AGEs Activities of a Novel Propolis Ultrafiltrate.

Propolis is one functional supplement with hundreds of years of usage. However, it's rarely consumed directly for its resinous property. Herein, a pre-treated process which can remove the impurity while preserve its bioactivities is needed to maximise its therapeutic opportunities. In the present study, a membrane-based ultrafiltration process was developed on a KM1812-NF experimental instrument. Using Brazilian green propolis as testing material, all experimental steps and parameters were sequentially optimized. In addition, a mathematical model was developed to fit the process. As a result, the optimum solvent was 60 % ethanol adjusted to pH 8-9, while the optimum MWCO (molecular weight cut-off) value of membrane was 30 KDa. The membrane filtration dynamic model fitted with the function y=(ax+b)/(1+cx+dx2 ). The resulting propolis ultrafiltrate from Brazilian green propolis, termed P30K, contains the similar profile of flavonoids and phenolic acids as raw propolis. Meanwhile, the ORAC (oxygen radical absorbance capacity) value of P30K is 11429.45±1557.58 µM TE/g and the IC50 value of inhibition of fluorescent AGEs (advanced glycation end products) formation is 0.064 mg/mL. Our work provides an innovative alternative process for extraction of active compounds from propolis and reveals P30K as an efficient therapeutic antioxidant.

The landscape of PD-L1 expression and somatic mutations in hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is the most common primary malignancy of the liver, and becoming the third-leading cause of cancer-related mortality worldwide. Despite the immune checkpoint inhibitors and molecular targeted therapies have shown preferable efficacy in HCC, large number of HCC patients do not respond effectively to anti-PD-1 reagents. Besides, the accumulation of genetic mutations in cancer cells may lead to the therapy resistant. Hence, there are clinical gaps between genetic and transcriptomic biomarkers for the HCC treatment. METHODS: To investigate the genetic mapping of liver cancer, targeted deep sequencing (TDS) and bioinformatics analysis were performed on hepatocellular carcinoma (HCC) tumor tissues and matched blood samples. Furthermore, copy number variants (CNVs) and Tumor mutation burden (TMB) were calculated. Immunohistochemistry was applied to determine the PD-L1 expression in HCC tumor tissues. Clinical characteristic, PD-L1 expression, and the TMB were analyzed in 32 HCC patients. RESULTS: This study indicated that the PD-L1 positive patients exhibited a lower TMB compared to the PD-L1 negative group, and PD-L1 positive patients were more likely to suffer from aggressive clinicopathologic features than PD-L1 negative patients. We also verified the top 30 mutated genes, including TP53, CTNNB1, KMT2D, AXIN1, ALK, and NOTCH1, in our dataset. Our results indicated that PD-L1 positive patients possessed more tumors with vascular invasion and advanced CCLC stage. Moreover, PD-L1 positive patients exhibited a lower TMB compared to the PD-L1 negative group. CONCLUSIONS: These findings could improve our understanding of the effects of immune checkpoint therapies on prognosis, and could facilitate the monitoring of somatic mutations in HCC.

Identify potential clinical significance of long noncoding RNA forkhead box P4 antisense RNA 1 in patients with early stage pancreatic ductal adenocarcinoma.

Previous studies have shown that forkhead box P4 antisense RNA 1 (FOXP4-AS1) is dysregulated in tumor tissues and can serve as a prognostic indicator for multiple cancers. However, the clinical significance of FOXP4-AS1 in pancreatic ductal adenocarcinoma (PDAC) remains unclear. The goal of this study is to recognize the possible clinical significance of long noncoding RNA FOXP4-AS1 in patients with early stage PDAC. A total of 112 patients from The Cancer Genome Atlas (TCGA) PDAC cohort, receiving RNA sequencing, were involved in the study. Survival analysis, functional mechanism, and potential small molecule drugs of target therapy of FOXP4-AS1 were performed in this study. Survival analysis in TCGA PDAC cohort suggested that patients with high FOXP4-AS1 expression had significantly augmented possibility of death than in PDAC patients with lower FOXP4-AS1 expression (adjusted P = .008; adjusted HR = 2.143, 95% CI = 1.221-3.760). In this study, a genome-wide RNA sequencing dataset was used to identify 927 genes co-expressing with FOXP4-AS1 in PDAC tumor tissues. A total of 676 differentially expressed genes were identified between different FOXP4-AS1 expression groups. Functional enrichment analysis of these genes and gene set enrichment analysis for PDAC genome-wide RNA sequencing dataset was done. We have found that FOXP4-AS1 may function in PDAC by participating in biological processes and pathways including oxidative phosphorylation, tricarboxylic acid cycle, classical tumor-related pathways such as NF-kappaB as well as Janus kinase/signal transducers in addition to activators of transcription, cell proliferation, and adhesion. In addition, we also screened two potential targeted therapeutic small molecule drugs (dimenhydrinate and metanephrine) for FOXP4-AS1 in PDAC. In conclusion, our present study demonstrated that higher expression of FOXP4-AS1 in PDAC tumor tissues were related with an inferior medical outcome. Through multiple genome-wide approaches, we identified the potential molecular mechanisms of FOXP4-AS1 in PDAC and two targeted therapeutic drugs for it.

Echo-Tracking Method Improves the Reliability in Flow-Mediated Dilation Assessment by Measuring Brachial Artery Reactivity.

The purpose of the study was to compare the intraobserver and interobserver reliability and agreement for measurement of flow-mediated dilation (FMD) between B-mode method and echo-tracking (ET) method. Twenty healthy volunteers (mean age, 31.6 ± 9.2 years) underwent ultrasound examination by both B-mode and ET methods. Baseline brachial artery diameter, post-cuff release diameter, and FMD percent were assessed by each sonologist on 2 consecutive days. Reliability was assessed by intraclass correlation coefficients, and Bland-Altman plots were used to visually compare measurement bias and agreement by the 2 ultrasound methods. A total of 40 pairs of data were available for analysis. Excellent intraobserver and interobserver reliability values were found for all variables assessed by the 2 methods. The intraclass correlation coefficient values were higher for ET in both intraobserver and interobserver evaluations, but only for interobserver evaluations for post-cuff release diameter and FMD was there no overlap in the 95% confidence interval. The Bland-Altman plots showed that in 95% of the measurements, the percentage difference between ET and B-mode ultrasound techniques was within 18.1%, 19.4%, and 17.3% for baseline brachial artery diameter, post-cuff release diameter, and FMD percent, respectively. The results suggest that ET and B-mode methods are reproducible in assessing the FMD. The ET method improves the reliability of FMD assessment, but we cannot determine which measurement is better for FMD.

[Role of CD44 in monocyte transmigration across Cryptococcus neoformans-infected blood-brain barrier in vitro].

OBJECTIVE: To explore the role of CD44 in monocyte adhesion to human brain microvascular endothelial cells (HBMECs) and monocyte migration across an in vitro model of blood-brain barrier (BBB) infected by Cryptococcus neoformans (Cn). METHODS: An in vitro blood-brain barrier model was constructed using a transwell chamber covered with a HBMEC monolayer. The wild-type strain of Cn B4500FO2, TYCC645#32 strain with CPS1 gene deletion and PCIP strain with CPS1 complementation were chosen to infect the monolayer HBMECs. THP-1 cells were added to the upper chamber of transwell, and the relative migration rate was determined by counting the number of the cells entering the lower chambers. The inhibitory effects of anti-CD44 monoclonal antibody and the CD44 inhibitor bikunin were examined on THP-1 binding to and migration across HBMECs. RESULTS: Cn infection of the HBMECs caused markedly enhanced THP-1 cell adhesion and migration across the monolyers (P<0.01) dependent on Cn concentration and exposure time. Addition of anti-CD44 monoclonal antibody and bikunin significantly lowered THP-1 adhesion and migration rates in the BBB model with Cn-infected HBMECs (P<0.01) with a dose dependence of the antibody (within 0-1 µg) and inhibitor (within 0-20 nmol/L). Both THP-1 adhesion rate and migration rate were lowered in the BBB model infected with CPS1 gene-deleted Cn but increased in the model infected with the complemented strain compared with those in the wild-type strain-infected model. CONCLUSION: In the in vitro BBB model, CD44 expressed on HBMECs may play an essential role in monocyte adhesion to and migration across the BBB. The capsular hyaluronic acid may mediate Cn-induced monocyte adhesion and migration.

Effect of Chinese herbal compound on liver fibrosis in rabbits with schistosomiasis by B-ultrasound.

OBJECTIVE: To explore the value of B-ultrasound on the evaluation of the effects of traditional Chinese medicine compound of Radix astragali, Salvia miltiorrhiza and Angelica sinensis, and TCM + praziquantel on liver fibrosis in rabbits with schistosomiasis. METHODS: The hepatic fibrosis model in rabbits with schistosomiasis was established. The experimental animals (24 rabbits) were randomly divided into four groups (group A, B, C and D, n=6). Group A (control group) was only treated by praziquantel; Group B was treated by mixture of Radix astragali and Salvia miltiorrhiza + praziquantel; Group C was treated by mixture of Radix astragali and Angelica sinensis + praziquantel; Group D was treated by mixture of Radix astragali, Salvia miltiorrhiza and Angelica sinensis + praziquantel. Then B-ultrasonogram was used to evaluate the effects. RESULTS: Each group showed certain curative effect on liver fibrosis in rabbits with schistosomiasis. The efficacy of group B, C and D was better than group A, and that of group D was the best. The differences in long diameter, thickness diameter, transverse diameter and portal vein inner diameter of liver before and after treatment were statistically significant (P<0.05). The liver function indexes and liver fibrosis indexes were significantly improved after treatment (P<0.05). CONCLUSIONS: The mixture of Radix astragali, Salvia miltiorrhiza and Angelica sinensis combined with Western medicine treatment can obviously improve the efficacy on liver fibrosis of schistosomiasis.

[Clinical features of chronic epididymitis: report of 63 cases].

OBJECTIVE: To summarize the clinical features of chronic epididymitis (CE) for the purpose of improving its diagnosis and treatment. METHODS: According to the specific inclusion and exclusion criteria, we selected 63 CE patients in this study, obtained the data on their symptoms, signs, sexual activities, histories of related diseases, impact on quality of life and CE symptom indexes (CESI) by interrogation, physical examination and questionnaires, assessed their correlation with CE, and summarized the clinical features of the disease. RESULTS: The case group showed a similarity to the controls in age, ethnicity, education, smoking and drinking, but significantly larger numbers of sexual partners and patients with a history of urinary tract infections than the latter. Epididymal swelling and tenderness were found in 92.1%, and scrotal pain in 75.5% of the CE patients. CESI and the score of the impact on quality of life were 7.9 +/- 4.6 and 12.5 +/- 5.6 in the case group, significantly higher than in the controls (4.4 +/- 3.2 and 8.5 +/- 4.2) (P<0.05). CONCLUSION: The significant signs of chronic epididymitis are epididymal swelling and tenderness, which affect the patient's quality of life. The association of chronic epididymitis with the number of sexual partners and history of urinary tract infections are yet to be further confirmed by larger-sample studies.

[Toxoplasma infection in males with sterility in Shenyang, China].

OBJECTIVE: To investigate the Toxoplasma gondii (TOX) infection in males with sterility and the effect of the infection on the reproductive function of males. METHODS: Enzyme linked immunoabsorbent assay (ELISA) was used to detect TOX-CAg, TOX-IgG and TOX-IgM in the peripheral blood of male patients with sterility. RESULTS: Among 100 cases of male sterility, 7 were TOX-IgG positive (7%), 16 TOX-IgM positive (16%) and 13 TOX-CAg positive (13%). Among 100 normal males, 7 were TOX-IgG positive (7%), 3 TOX-IgM positive (3%) and 1 TOX-CAg positive (1%). CONCLUSION: TOX infection may affect the fertility of males and cause male sterility. For this reason, males should prevent themselves from TOX infection.