Research progress in strigolactones and application prospect in medicinal plants / 中国中药杂志

Strigolactones(SLs) are a class of sesquiterpenoids derived from the carotenoid biosynthesis pathway with the core carbon skeleton consisting of tricyclic lactone(ABC tricyclic ring) and α,β-unsaturated furan ring(D ring). SLs are widely distributed in higher plants and are symbiotic signals between plants and Arbuscular mycorrhiza(AM), which play key roles in the evolution of plant colonizing terrestrial habitats. As a new type of plant hormone, SLs possess such important biological functions as inhibiting shoot branching(tillers), regulating root architecture, promoting secondary growth, and improving plant stress resistance. Therefore, SLs have attracted wide attention. The biological functions of SLs are not only closely related to the formation of "excellent shape and quality" of Chinese medicinal materials but also have important practical significance for the production of high-quality medicinal materials. However, SLs have been currently widely studied in model plants and crops such as Oryza sativa and Arabidopsis thaliana, and few related studies have been reported on SLs in medicinal plants, which need to be strengthened. This review focused on the latest research progress in the isolation and identification, biological and artificial synthesis pathways, biosynthesis sites and transport modes, signal transduction pathways and mechanisms, and biological functions of SLs, and prospected the research on the regulation mechanism of SLs in the growth and development of medicinal plants and their related application on targeted regulation of Chinese herbal medicine production, which is expected to provide some references for the in-depth research on SLs in the field of Chinese medicinal resources.

Fatty acid quantification method by isobutyl esterification and GC-FID/MS / 上海交通大学学报（医学版）

Objective:To establish a quantification method for understanding the varieties and concentrational changes of fatty acids under physiological and pathophysiological conditions. Methods:Based on isobutyl esterification using (-)-menthyl chloroformate and isobutanol, 27 typical fatty acids were qualitatively and quantitatively analyzed by gas chromatography-flame ionization detector/mass spectrometry (GC-FID/MS). The sensitivity and stability of the method were detected by limit of detection (LOD), limit of quantification (LOQ), and intra-day and inter-day relative standard deviation (RSD). The method was applied to four typical biological samples (human serum, urine, feces and rat liver) to verify the universality in different substrates. Results:Isobutyl esters of 27 fatty acids were effectively separated with an HP-5MS column. The results showed nice linearity within 2-3 orders of magnitude in terms of concentration with R2>0.99 for all 27 tested fatty acids. The LODs of the method were between 0.03 and 2.96 pmol on column whereas the LOQs were between 0.09 and 9.86 pmol. The results of method validation showed that the intra-day and inter-day RSDs were all under 10% in the range of high, medium and low concentrations. With this method, moreover, 10, 7, 14 and 9 fatty acids were detected in human serum, urine, feces and rat liver, respectively. Conclusion:A quantitative analytical method for fatty acids with short, media and long chains (C1-C24) is established based on isobutyl estrification in aqueous media. This method has the advantages of mild condition, high sensitivity, good stability, and simple and quick operation, and can be directly used in the detection of serum, urine, feces, liver and other biological samples, which may provide a new idea for high-throughput metabolome analysis.

Effect of atorvastatin on inflammatory infiltration in the lung of rabbits with hypercholesterolemia / 中南大学学报(医学版)

Objective To determine the effect of atorvastatin on the hypercholesterolemia in-duced lesion in the lung. Methods Fifteen male New Zealand rabbits were randomly assigned into a control group (n=5) , a high-cholesterol forage group (n=5) , and an atrovastatin treatment group (n=5). The control group received normal forage, but the high-cholesterol group and atrovastatin treatment group received high-cholesterol forage. From the 9 th week, the atrovastatin treatment group was added atorvastatin, and the experiment stopped at the end of the 14th week. At the beginning of the experiment and at the 8 th, 14 th week, blood cholesterol and body weight were detected. At the 14th week, bronchial alveolar lavage (BAL) was performed in vitro after the rabbits were executed; pathological examinations were determined in the lung tissues by staining with hamatoxylin-eosin. Oil red 0 and the activities of NF-κB in the alveolar macrophages (AMs) were investigated by immuno-cytochemistry. Proliferative cell nuclear antigen in the lung tissues was adopted by immunohistochem-istry, and the concentrations of IL-6 in the serum, BALF and the culture supematants of AMs were measured by ELISA. Pulmonary tissue paraffin section was stained with hamatoxylin-eosin. Results Atorvastatin reduced inflammatory infiltration, AM NF-κB activation, and cell proliferation in the lung, but raised IL-6 level. Conclusion Hypercholesterolemia-induced pulmonary inflammation is attenuated by atorvastatin.

Immune responses against Schistosoma japonicum after vaccinating mice with a multivalent DNA vaccine encoding integrated membrane protein Sj23 and cytokine interleukin-12 / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The vaccination of mice with DNA encoding single candidate antigens has failed to induce significant protection against Schistosoma japonicum (S. japonicum) challenge infections. In this study, we evaluated the feasibility of using a multivalent DNA vaccine which co-expressed S. japonicum integral membrane protein Sj23 and murine cytokine IL-12 to induce protective immune responses.</p><p><b>METHODS</b>The plasmid pVIVO2-IL12-Sj23, a eukaryotic expression vector expressing Sj23 and murine IL-12 simultaneously, was constructed, identified, and tested for expression in vitro. Its ability to protect against S. japonicum challenge infections was analyzed according to worm reduction rate and egg reduction rate after vaccination of BALB/c mice. The serum levels of specific IgG antibody were determined by enzyme-linked-immuno sorbent assay (ELISA) and Western blot analysis. Using cultured spleen cells, IFN-gamma and IL-4 post-stimulation were quantified by ELISA. The phenotypes of splenocyte populations were analyzed by flow cytometry (FCM).</p><p><b>RESULTS</b>The plasmid DNA pVIVO2-IL12-Sj23 was proven to express well in vitro by transient transfection of HEK-293 cells. Immunization resulted in a worm reduction rate of 45.53% and egg reduction rate of 58.35%. ELISA and Western blot analysis indicated that immunized mice generated specific IgG against Sj23. Spleen cells showed significant increases in IFN-gamma but decreases in IL-4. No significant differences in CD4+ and CD8+ subgroup ratios were observed after the challenges.</p><p><b>CONCLUSIONS</b>The multivalent DNA vaccine pVIVO2-IL12-Sj23 is sufficient to elicit moderate but highly significant levels of protective immunity against challenge infections. Cytokine IL-12, as a gene adjuvant, was able to enhance the Th1 responses and, hence, the protective immunity.</p>

Congenital infection of rabbits with Schistosoma japonicum and protective immunity of offspring / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Recently congenital infection with Schistosoma japonicum (S. japonicum) has been demonstrated in pigs, rabbits, mice and dogs. We explored the rabbit as an animal model for the congenital infection of schistosomiasis japonica and assessed the effect of a congenital S. japonicum infection on the resistance of rabbit kittens to a postnatal challenge infection.</p><p><b>METHODS</b>Sixteen pregnant New Zealand white rabbits were infected with a single dose of S. japonicum cercariae. The exposed animals were divided into three groups according to the gestation age at the time of infection. Diagnosis of prenatally acquired S. japonicum infection in the rabbit kittens was primarily based on serological tests in combination with parasitological and histopathological findings. Congenitally infected kittens were challenged percutaneously with 100 S. japonicum cercariae to assess the effect of a congenital S. japonicum infection on kitten resistance to a postnatal challenge infection.</p><p><b>RESULTS</b>The overall prevalence of congenital infection in offspring of infected mothers was 20% (12/60). The congenital infection rate in group L (late gestation) was much higher than in group E (early gestation) and group M (mid-gestation) (P <0.05). After a postnatal challenge infection, prenatally infected kittens had a 54.66% worm reduction rate, 41.45% egg reduction rate, and 51.76% granuloma size reduction rate compared to naïve kittens.</p><p><b>CONCLUSIONS</b>This study demonstrates the possibility of congenital infection of S. japonicum in rabbits and the resistance of congenitally infected kittens to a postnatal challenge infection. These results have important implications not only for epidemiological investigations, but also in designing government control programs for schistosomiasis.</p>