Protection of the liver against CCl_4-induced injury by intramuscular electrotransfer of a kallistatin-encoding plasmid

AIM:To investigate the effect of transgenic expression of kallistatin(Kal) on carbon tetrachloride(CCl 4)induced liver injury by intramuscular(im) electrotransfer of a Kal-encoding plasmid formulated with poly-Lglutamate(PLG).METHODS:The pKal plasmid encoding Kal gene was formulated with PLG and electrotransferred into mice skeletal muscle before the administration of CCl 4.The expression level of Kal was measured.The serum biomarker levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),malonyldialdehyde(MDA),and tumor necrosis factor(TNF)-α were monitored.The extent of CCl 4-induced liver injury was analyzed histopathologically.RESULTS:The transgene of Kal was sufficiently expressed after an im injection of plasmid formulated with PLG followed by electroporation.In the Kal gene-transferred mice,protection against CCl 4-induced liver injury was reflected by significantly decreased serum ALT,AST,MDA and TNF-α levels compared to those in control mice(P < 0.01 to 0.05 in a dose-dependent manner).Histological observations also revealed that hepatocyte necrosis,hemorrhage,vacuolar change and hydropic degeneration were apparent in mice after CCl 4 administration.In contrast,the damage was markedly attenuated in the Kal gene-transferred mice.The expression of hepatic fibrogenesis marker transforming growth factor-β1 was also reduced in the pKal transferred mice.CONCLUSION:Intramuscular electrotransfer of plasmid pKal which was formulated with PLG significantly alleviated the CCl 4-induced oxidative stress and inflammatory response,and reduced the liver damage in a mouse model.

Inhibition of angiogenesis and HCT-116 xenograft tumor growth in mice by kallistatin

AIM: To investigate the inhibitory effect of kallistatin (KAL) on angiogenesis and HCT-116 xenograft tumor growth. METHODS: Heterotopic tumors were induced by subcutaneous injection of 2 × 106 HCT-11 cells in mice. Seven days later, 2 × 1011 rAAV-GFP or rAAV-KAL was injected intratumorally (n = 5 for each group). The mice were sacrificed at d 28, by which time the tumors in the rAAV-GFP group had grown to beyond 5% of the total body weight. Tumor growth was measured by calipers in two dimensions. Tumor angiogenesis was determined with tumor microvessel density (MVD) by immunohistology. Tumor cell proliferation was assessed by Ki-67 staining. RESULTS: Intratumor injection of rAAV-KAL inhibited tumor growth in the treatment group by 78% (171 ± 52 mm3) at d 21 after virus infection compared to the control group (776 ± 241 mm3). Microvessel density was significantly inhibited in tumor tissues treated with rAAV-KAL. rAAV-KAL also decreased the proportion of proliferating cells (Ki-67 positive cells) in tumors compared with the control group.CONCLUSION: rAAV-mediated expression of KAL inhibits the growth of colon cancer by reducing angiogenesis and proliferation of tumor cells, and may provide a promising anti-angiogenesis-based approach to the treatment of metastatic colorectal cancer.

Chemical Composition of the Fatty Oil from Fructus Broussonetiae and Its Effects on Rat Plasma Lipids and Adipose Tissue

Context: Broussonetia papyrifera (L.) Vent. (Moraceae), a traditional Chinese medicinal herb, has been extensively applied for many years to treat various diseases. Its fruits (Fructus Broussonetiae) have been commonly used as an important tonic for the treatment of age-related disorders with long history;recent research has proved that it contains 32% to 35% fixed oils. The fixed oil is composed mainly of unsaturated fatty acids, including linoleic acid, methyl palmitate, oleic acid and linoleic acid ester. Objective: To investigate the chemistry of the fatty oil from Fructus Broussonetiae (FOFB) and its effects on plasma lipids. Methods: The chemical composition of FOFB was examined and identified by GC-MS. Thirty male Wistar rats fed diet containing FOFB and cholesterol were studied for 28 days. The effect of dietary FOFB on plasma lipids and adipose tissue was tested. Results: Twelve compounds of FOFB were examined and identified, the major components of fatty oil, 8,11-octadecadienoic acid (83.75%), palmitic acid (10.22%), octadecadienoic acid (2.97%) and 9-octadecenoic acid (1.69%) were found. FOFB significantly exhibited the activities of decreasing the rat adipose tissue weight, triacylglycerol, total cholesterol, and low-density lipoprotein cholesterol (LDL-C) concentrations while the rat body weight remained unchanged. Discussion: FOFB contained a large amount of PUFA which had the effect on reducing plasma lipids and adipose.

Salvianolic acid B increases recombinant adeno-associated virus transduction through redirecting trafficking pathways and stabilizing perinuclear accumulations

OBJECTIVE To screen small molecule compounds from traditional Chinese medicine that can enhance recombinant adeno-associated virus(rAAV) transduction.METHODS Recombinant adeno-associated virus(rAAV) has been established as a powerful tool for in vivo gene transfer and achieved much promise in gene therapy applications.However,widespread clinical use has been limited by transduction efficiency.In the current study,we screened a panel of small molecule compound from traditional Chinese medicine focused on AAV intracellular trafficking process and found salvianolic acid B can significantly enhance rAAV2 transduction.RESULTS Salvianolic acid B caused a dose-depen.dent increase in rAAV2 transduction regardless of vector dose,genome architecture,and over a broad range of cell line from various cell type and species(HEK293,HeLa,HepG2,Huh-7,CHO-K1,LO-2).Salvianolic acid B treatment redirected rAAV2 particles toward large vesicles positive for late endosomal(Rab7) and lysosomal(LAMP1) markers.Furthermore,salvianolic acid B acted to increase accumulation of viral particles at the perinuclear region.CONCLUSION In summary,our results suggest that salvi.anolic acid B redirects rAAV2 toward more productive trafficking pathways and stabilizes perinuclear accumulations of vectors,facilitating productive nuclear trafficking.

Associations between the use of red yeast rice preparations and adverse health outcomes:An umbrella review of meta-analyses of randomized controlled trials

Background:Red yeast rice(RYR),a natural lipid-lowering agent,is widely used in clinical practice.However,the existing meta-analyses concerning the safety of RYR preparations have yielded inconsistent results,and the credibility of the evidence has not been quantified.Objective:This study was designed to evaluate the existing evidence and offer a comprehensive understanding of the associations between the use of RYR preparations and various adverse health outcomes.Search strategy:Seven literature databases were searched from inception to May 5,2023,using medical subject headings and free-text terms(e.g.,“red yeast rice,”“Xuezhikang,”and“Zhibitai”).Inclusion criteria:Meta-analyses that investigated and quantitatively estimated associations between the use of RYR preparations and adverse health outcomes were included in this study.Data extraction and analysis:Two researchers independently extracted data using a standardized data collection table;any disagreements were resolved by consulting a third researcher.Based on the participant,intervention,comparator and outcome(PICO)framework in each eligible meta-analysis,a series of unique associations between the use of RYR preparations and adverse health outcomes were determined.The associations’effect estimates were re-evaluated using random-effect models.Results:Fifteen meta-analyses,comprising 186(164 unique)randomized controlled trials,were identified.Based on A Mea Surement Tool to Assess Systematic Reviews version 2,3(20%)and 12(80%)of these meta-analyses had low and critically low confidence,respectively.A total of 61 unique associations between the use of RYR preparations and adverse health outcomes were extracted from eligible metaanalyses.Based on the random-effect models,10(16.4%)associations indicated a significant protective effect of RYR preparations against adverse health outcomes,while 5(8.2%)indicated an increased risk of adverse health outcomes related to uric acid,alanine transaminase and aspartate transaminase levels.The other 46(75.4%)associations showed no significant difference between the use of RYR preparations and control treatments.Regarding the credibility of the evidence,21(34.4%),34(55.7%)and 6(9.8%)associations showed moderate,low and very low credibility,respectively.Conclusion:The evidence examined in this study suggests that RYR preparations are safe;however,the credibility of the evidence was not high.Further high-quality evidence is required.

Laser speckle contrast imaging based on uniting spatiotemporal Fourier transform

We propose a laser speckle contrast imaging method based on uniting spatiotemporal Fourier transform.First,the raw speckle images are entirely transformed to the spatiotemporal frequency domain with a three-dimensional(3D)fast Fourier transform.Second,the dynamic and static speckle components are extracted by applying 3D low-pass and high-pass filtering in the spatiotemporal frequency domain and inverse 3D Fourier transform.Third,we calculate the time-averaged modulation depth with the average of both components to map the two-dimensional blood flow distribution.The experiments demonstrate that the proposed method could effectively improve computational efficiency and imaging quality.