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OBJECTIVE@#Melittin, a cell-penetrating peptide, improves the efficiency of many non-viral gene delivery vectors, yet its application in viral vectors has not been well studied. The non-pathogenic recombinant adeno-associated virus (rAAV) vector is an ideal in vivo gene delivery vector. However, its full potential will only be achieved after improvement of its transduction efficiency. To improve the transduction efficiency of rAAV2 vectors, we attempted to develop a melittin-based rAAV2 vector delivery strategy.@*METHODS@#The melittin peptide was inserted into the rAAV2 capsid either in the loop VIII of all viral proteins (VPs) or at the N terminus of VP2. Various rAAV2-gfp or -fluc vectors were subjected to quantitative real-time polymerase chain reaction and Western blot assays to determine their titers and integrity of capsid proteins, respectively. Alternatively, the vectors based on wild-type capsid were pre-incubated with melittin, followed by transduction of cultured cells or tail vein administration of the mixture to C57BL/6 and BALB/c nude mice. In vivo bioluminescence imaging was performed to evaluate the transgene expression.@*RESULTS@#rAAV2 vectors with melittin peptide inserted in the loop VIII of VPs had low transduction efficiency, probably due to dramatically reduced ability to bind to the target cells. Fusing the melittin peptide at the N-terminus of VP2 produced vectors without the VP2 subunit. Interestingly, among the commonly used rAAV vectors, pre-incubation of rAAV2 and rAAV6 vectors with melittin significantly enhanced their transduction efficiency in HEK293 and Huh7 cells in vitro. Melittin also had the ability to increase the rAAV2-mediated transgene expression in mouse liver in vivo. Mechanistically, melittin did not change the vector-receptor interaction. Moreover, cell counting kit-8 assays of cultured cells and serum transaminase levels indicated melittin had little cytotoxicity.@*CONCLUSION@#Pre-incubation with melittin, but not insertion of melittin into the rAAV2 capsid, significantly enhanced rAAV2-mediated transgene expression. Although further in vivo evaluations are required, this research not only expands the pharmacological potential of melittin, but also provides a new strategy to improve gene therapy mediated by rAAV vectors.
Subject(s)
Mice , Animals , Humans , Melitten/genetics , Dependovirus/genetics , Serogroup , HEK293 Cells , Mice, Nude , Mice, Inbred C57BL , Transgenes , Genetic Vectors/geneticsABSTRACT
OBJECTIVE@#High-fat diet is one of the main risk factors that disrupt the balance of gut microbiota, which eventually will induce colorectal cancer (CRC). Evodiamine (EVO) is a wildly used multifunctional traditional Chinese medicine extract. In this study, we investigated the role of gut microbiota in high-fat diet-propelled CRC and the potential of EVO for CRC chemoprevention.@*METHODS@#Gut microbiota, serum d-lactic acid and endotoxin from 38 patients with colon cancer and 18 healthy subjects were detected by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay (ELISA). In addition, body mass index, phospho-signal transducer and activator of transcription 3 (p-STAT3) expression in cancer tissues and paracancerous tissues were detected by immunohistochemistry. A mouse intestinal inflammatory tumor model was established by azomethane/sodium dextran sulfate, followed by treatment with EVO and 5-aminosalicylic acid (ASA). Gut microbiota and inflammatory factors were detected by quantitative polymerase chain reaction, while serum d-lactic acid and endotoxin were detected by ELISA. Furthermore, cell proliferation, cell apoptosis, and interleukin (IL)-6/STAT3/P65 pathway were evaluated by 5-ethynyl-2'-deoxyuridine, terminal-deoxynucleotidyl transferase-mediated nick-end labeling, and Western blot assays.@*RESULTS@#In patients with colon cancer, the numbers of Enterococcus faecalis and Escherichia coli were increased, while those of Bifidobacterium, Campylobacter and Lactobacillus were decreased. Serum endotoxin and d-lactic acid levels and p-STAT3 levels were significantly increased. In the mouse model, both EVO and ASA inhibited tumor formation, decreased the proliferation of tumor cells, and induced apoptosis of tumor cells. Compared with the control group, the numbers of E. faecalis and E. coli were decreased, while Bifidobacterium, Campylobacter and Lactobacillus numbers were increased. In the EVO group, serum endotoxin and d-lactic acid levels and inflammatory factors were significantly decreased. Further, the IL6/STAT3/P65 signaling pathway was inhibited in the EVO group.@*CONCLUSION@#EVO may inhibit the occurrence of colon cancer by regulating gut microbiota and inhibiting intestinal inflammation. The potential mechanism involves inhibition of the IL6/STAT3/P65 signaling pathway, revealing its potential therapeutic significance in clinical applications.
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Objective: To detect the expression of peroxisome proliferator-activated receptor (PPAR) in ovarian tissues of rats with chronic low-grade inflammation, and to explore the effect of PPAR-γ agonist rosiglitazone on ovarian dysfunction induced by chronic low-grade inflammation. Methods: A chronic low-grade inflammation model of rats was established by intraperitoneal injection of lipopolysaccharide (LPS). Two hundred rats were randomly assigned to control group (NS group) and chronic low-grade inflammation group (LPS group), and the rats were intraperitoneally injected with normal saline and LPS, respectively. The ovarian function of rats was assessed by detecting the serum levels of estradiol (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH), and anti-Müllerian hormone (AMH) during different stages of the estrus cycle (proestrus, estrus, metestrus and diestrus). The protein expression levels of PPAR-α, PPAR-γ and PPAR-γ in ovarian tissues were detected using Western blotting. Eighty rats of each group were randomly divided into two subgroups, which were administered intragastrically by normal saline and rosiglitazone, respectively. Fourteen days after intragastric administration, the levels of proinflammatory cytokines, including interleukin (IL)-1β, IL-6 and tumor necrosis factor α (TNF-α), in ovarian tissues and ovarian function of rats in each subgroup were observed during different stages of the estrus cycle. Results: Compared with the NS group, during different stages of the estrus cycle, the serum levels of E2 and AMH of rats in the LPS group were significantly decreased (all P < 0.05), while the serum levels of FSH and LH were significantly increased (all P < 0.05). During different stages of the estrus cycle, the expression levels of PPAR-γ in ovarian tissues were significantly decreased in the LPS group compared with the NS group (all P < 0.05), while the expression levels of PPAR-α and PPAR-δ were not significantly different between the two groups (all P < 0.05). Compared with the intraperitoneal injection of LPS intragastric administration of normal saline subgroup, during different stages of the estrus cycle, the expression levels IL-1β, IL-6 and TNF-α in ovarian tissues of rats were significantly decreased in the intraperitoneal injection of LPS intragastric administration of rosiglitazone subgroup (all P < 0.05), the serum levels of E2 and AMH were significantly increased (all P < 0.05), and the serum levels of FSH and LH were significantly decreased (all P < 0.05). Conclusion: PPAR-γ agonist rosiglitazone can attenuate LPS-induced chronic low-grade inflammatory and improve ovarian function in rats.
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Objective To study the pregnancy complication incidences, delivery manners and pregnancy outcomes of senile pregnant women. Methods The medical records of 10 508 cases of pregnant and delivery women, who were hospitalized for delivery in Obstetrical Department of Changhai Hospital of Second Military Medical University from Jan. 2010 to Dec. 2016, were collected and analyzed. According to delivery age, they were divided into the senile group (≥35 years old, n＝1 130) and the non-senile group (35 years old, n＝9 378). Chi-square test was used to compare the incidences of pregnancy complications, cesarean section rate, ratio of cesarean section indications and outcomes of maternal and perinatal infants between the two groups. Results The proportion of senile pregnant women increased from 6.75% (89/1 319) in 2010 to 15.82% (267/1 688) in 2016. The cesarean section rate and the ratio of social factors in the senile group were higher than those in the non-senile group (67.08% [758/1 130] vs 48.51% [4 549/9 378], 42.88% [325/758] vs 10.07% [458/4 549], both P0.01). The incidences of gestational diabetes mellitus, premature rupture of membrane, placenta praevia, placental implantation abnormality and postpartum hemorrhage of the senile group were significantly higher than those of the non-senile group (all P0.05). The incidences of pregnancy-induced hypertension syndrome, pre-eclampsia, chronic hypertension complicating pregnancy of the senile group were significantly higher than those of the non-senile group (all P0.05). The incidences of liver diseases, diseases of urinary system, abnormal thyroid function, thrombotic diseases and malignant tumor complicated with pregnancy were significantly higher in the senile group than those in the non-senile group (all P0.05). For outcome of perinatal infants, the onset rates of asphyxia neonatorum, death of perinatal infants and premature infants were significantly higher in the senile group than those in the non-senile group (all P0.05). Conclusion The risks of pregnancy complications and adverse pregnancy outcomes are increased in senile pregnant women. Appropriate-age pregnancy and perinatal care of senile pregnancy should be advocated in clinic.
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Anoectochilus roxburghii is a traditional Chinese medicine and natural health products. In the modern cultivation system, A. roxburghii is micropropagated in tissue culture, and the plants are transferred to soil cultivation for months. However, it remains unclear about the necessity of soil cultivation for the accumulation of health beneficial compounds. In this paper, we performed nontargeted metabolomic analysis using GC-TOF-MS and UPLC-Q-TOF-MS, on A. roxburghii plants at tissue culture stage or after 3 months of soil cultivation. The results showed that the primary metabolites such as alcohols and organic acids are abundant in the tissue culture plants. In contrast, polysaccharide, nucleoside, esters and secondary metabolites such as flavonoids, terpenoids were significantly accumulated in cultivated seedlings. Flavonoids and polysaccharides are considered as the principle effective components in A. roxburghii. Soil cultivation period is therefore essential for the accumulation of these metabolites.
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<p><b>BACKGROUND</b>It is the first multicenter clinical study in China to investigate zanamivir use among Chinese adolescents and adults with influenza-like illness (ILI) since 2009, when inhaled zanamivir (RELENZA(®)) was marketed in China.</p><p><b>METHODS</b>An uncontrolled open-label, multicentre study to evaluate the antiviral activity, and safety of inhaled zanamivir (as Rotadisk via Diskhaler device); 10 mg administered twice daily for 5 days in subjects ≥ 12 years old with ILI. Patients were enrolled within 48 hours of onset and followed for eight days. Patients were defined as being influenza-positive if the real-time reverse transcriptase-polymerase chain reaction (rRT-PCR) test had positive results.</p><p><b>RESULTS</b>A total of 400 patients ≥ 12 years old were screened from 11 centers in seven provinces from March 2010 to January 2011. Three hundred and ninety-two patients who took at least one dose of zanamivir were entered into the safety analysis. The mean age was 33.8 years and 50% were male. Cardiovascular diseases and diabetes were the most common comorbidities. All the reported adverse events, such as rash, nasal ache, muscle ache, nausea, diarrhea, headache, occurred in less than 1% of subjects. Mild sinus bradycadia or arrhythmia occurred in four subjects (1%). Most of the adverse events were mild and did not require any change of treatment. No severe adverse events (SAE) or fatal cases were reported. Bronchospasm was found in a 38 years old woman whose symptoms disappeared after stopping zanamivir and without additional treatment. All the 61 influenza virus isolates (43 before enrollment, 18 during treatment) proved to be sensitive to zanamivir.</p><p><b>CONCLUSIONS</b>Zanamivir is well tolerated by Chinese adolescents and adults with ILIs. There is no evidence for the emergence of drug-resistant isolates during treatment with zanamivir.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Antiviral Agents , Therapeutic Uses , Influenza, Human , Drug Therapy , Treatment Outcome , Zanamivir , Therapeutic UsesABSTRACT
Gas chromatography/mass spectrometry, chemometric resolution method-Alternative movingwindow factor analysis that were proposed recently and the Kovats retention index were used to analyze the essential components of herbal pair Cinnamomum cassia Presl and Poria cocos (CCP-PC) and compare them with those of single herbal Cinnamomum cassia Presl (CCP). 46 and 42 essential components in essential oil of CCP and CCP-PC have been identified individually. Results shows that the number of essential components of CCP and CCP-PC were almost the same, but extractive ratioes of them have changed significantly, some of them were increased obviously, most of them were declined notably instead and even several ingredients of CCP-PC were disappeared due to CCP PC's interaction probably. The main pharmacodynamic ingredients of CCP-PC, 3-phenyl-2-propenal and Cinnamaldehyde were obviously higher contents than that of single CCP. It suggested that there exist certain interactions of the chemical ingredients in compound medicine rather than their sum effect of single medicines. There is a litter difference in quality and quantity between single medicines and pair medicines, so the application of pair medicines can expand single medicine's adaptive disease and has a good clinical referenced valuation.
Subject(s)
Cinnamomum aromaticum , Chemistry , Drugs, Chinese Herbal , Chemistry , Gas Chromatography-Mass Spectrometry , Poria , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To analyze the status of liver disease research in China.</p><p><b>METHOD</b>Hepatology and gastroenterology journals in the Science Citation Index were searched and their impact factors were analyzed.</p><p><b>RESULTS</b>19 periodicals in this field were published in 6 countries. Among them 11 (61%) were published in the USA. Eleven of the 19 were full text embodied in the ScienceDirect (Elsevier), Blackwell and Wiley. The impact factor of the journal Gastroenterology was 13.092, the highest or the 19, and Hepatology, with an impact factor of 10.416, was in the second position. Of the 19 periodicals, 12 had an impact factor over 2.822. Articles published in those journals from 1001 through 2005 were written by mainland Chinese authors.</p><p><b>CONCLUSION</b>The 12 periodicals with an impact factor over 2 can be regarded as core periodicals in this field. The medical periodicals in China need to be further improved in order to reach top international standards.</p>