ObjectiveTo investigate the alteration of MexB protein expression by plasmid containing siRNA template strand was transformed into Pseudomonas aeruginosa.Methods siRNA molecules specifically against mexB were designed and ligated into pGPU6/GFP/Neo vector to construct the recombinant plasmid pGPU6/GFP/Neo-siRNA.MexB gene was cloned into expression vector pET22b+ to construct plasmid pET22b+/mexB,the recombinant expression plasmid was transformed into E.coli BL21 (DE3)plysS and protein MexB was induced to express,then purified protein MexB was used to prepare specific antibodies in rabbits.The plasmids with siRNA molecules specifically against mexB were transformed into wild type strain,clinical multiresistant strain and mexB overexpression strain of Pseudomonas aeruginosa by electroporation respectively,and the changes of the expression of MexB were detected in 8,12,24 h respectively by Western blot.ResultspGPU6/GFP/Neo-siRNA was constructed successfully.Protein MexB was expressed successfully and the rabbit polyclonal antibodies against MexB was prepared well.The gene silence of mexB by siRNA molecules was effective in the three strains of Pseudomonas aeruginosa,but it depended on the silencing time.ConclusionThe expression of MexB was reduced in 8 h and 12 h,but in 24 h,the expression was unchanged.

OBJECTIVEMany studies have demonstrated that low levels of insulin-like growth factor-I (IGF-I) may be associated with the hypoxic-ischemic brain damage (HIBD) and that IGF-I has a neuroprotective effect. The role of IGF-II, a structurally and functionally homologous polypeptide with IGF-I, is unclear in HIBD. This study was designed to observe the changes of serum and cerebrospinal fluid (CSF) IGF-II levels in neonates with hypoxic-ischemic encephalopathy (HIE) and to investigate its effects on HIE.

METHODSSerum and CSF IGF-II levels in 41 neonates with HIE were measured by radioimmunoassay in the acute phase (postnatal age 12-24 hrs) and the convalescence phase (postnatal age 10-12 days). The 41 HIE neonates included 10 cases of mild, 12 moderate, and 19 severe HIE. Serum samples of 10 normal neonates were used as controls.

RESULTSIn the acute phase, serum IGF-II levels in the Mild HIE group (203.28 +/- 40.09 ng/mL) and the Moderate HIE group (192.33 +/- 39.66 ng/mL) were not significantly reduced, but were obviously reduced in the Severe HIE group (116.72 +/- 39.50 ng/mL) compared with normal controls (229.38 +/- 43.39 ng/mL) (P<0.01). During the convalescence phase, serum IGF-II levels in the Mild HIE group (285.53 +/- 49.44 ng/mL) and in the Moderate HIE group (278.69 +/- 51.34 ng/mL) increased significantly (P < 0.01); CSF IGF-II levels increased in the Mild HIE group from 27.23 +/- 7.82 ng/mL (acute phase) to 81.58 +/- 9.77 ng/mL (convalescence phase) (P < 0.01) and also increased in the Moderate HIE group from 23.43 +/- 7.79 ng/mL (acute phase) to 78.48 +/- 10.44 ng/mL (convalescence phase) (P < 0.01). The patients from the severe HIE group whose neurological symptoms or signs were improved in the convalescence showed higher serum and CSF IGF-II levels than in the acute phase (254.08 +/- 48.50 ng/mL vs 122.21 +/- 46.26 ng/mL; 69.42 +/- 10.20 ng/mL vs 15.05 +/- 7.03 ng/mL; P < 0.01). A positive correlation was found between the serum and CSF IGF-II levels in the HIE group (r=0.69, P < 0.01).

CONCLUSIONSIGF-II levels in serum and CSF are associated with the pathogenesis and the prognosis of neonatal HIE.

Female ; Humans ; Hypoxia-Ischemia, Brain ; metabolism ; Infant, Newborn ; Insulin-Like Growth Factor II ; analysis ; cerebrospinal fluid ; Male

The pharmacokinetics of traditional Chinese medicine is a subject that studies the dynamic changes of the absorption, distribution, metabolism and excretion of complex components of traditional Chinese medicine, which is an important method for elucidating the pharmacodynamic material basis, action characteristics, and compatibility mechanisms of traditional Chinese medicine. However, given on the fact that traditional Chinese medicine is a multi-dimensional and complex system with multiple components, multiple pathways, multiple targets, and an unclear pharmacodynamic material basis, the research on the pharmacokinetics of traditio-nal Chinese medicine has become a scientific and technological problem. Although the pharmacokinetics of traditional Chinese medicine has achieved remarkable development with the emergence of new theories, methods and technologies, there are still some problems in the application of the research direction of the pharmacokinetics of traditional Chinese medicine judging from the current application of the National Natural Science Foundation of China. Therefore, this article discussed the current research status on pharmacokinetics of traditional Chinese medicines by analyzing the projects funded by the National Natural Science Foundation of China in the past 5 years from 2016 to 2020, mainly including the application and funding analysis, main research contents of the projects in pharmacokinetics of traditional Chinese medicines. And the research hotspots, difficulties and deficiencies were focused in order to provide certain reference for researchers engaged in pharmacokinetics of traditional Chinese medicine.
China ; Financial Management ; Foundations ; Medicine, Chinese Traditional ; Natural Science Disciplines