Bioinformatics analysis on structure and function and expression vector construction of SARS-CoV-2 related protein TMPRSS2 / 中国药理学通报

Aim Human TMPRSS2 is a transmembrane serine protease.In this paper, the structure and func¬tion of the protein were systematically analyzed by bioinformatics, the codon was optimized and the pro- karvotie expression vector was constructed to explore the molecular mechanism of SARS-CoV-2 infecting host cells.Methods The recombinant expression vector pET-22b-TMPRSS2 was generated by molecular clo¬ning technology.The homology, functional sites, sub¬cellular localization, three-dimensional structure and evolutionary characteristics of TMPRSS2 protein were systematically analyzed by using analytical tools such as Protparam, NetPhos3.1, Blast, Clustal X2 and MEGA7.0.Results The prokarvotic expression plas- mid was constructed correctly; TMPRSS2 belongs to medium molecular weight protein, which is composed of 492 amino acid residues.The theoretical isoelectric point is 8.12, the molecular extinction coefficient is 118 145 L • mol~1 • cm"1 , and the half-life is 30 h; TMPRSS2 has 15 potential glycosylation sites and 49 possible phosphorylation sites.It is a transmembrane hydrophilie protein without signal sequenee.In addi¬tion, the protein has 13 potential B-cell epitopes and 7 T-eell epitopes.Seeondarv structure analysis showed that random coil accounted for the highest proportion of TMPRSS2 protein ( 0.453 3) , followed by extended strand (0.252 0).Sequence comparison and evolu¬tionary analysis showed that the highest sequence con¬sistency and closest genetic relationship with human TMPRSS2 was Pan troglodytes, followed by gorilla.Conclusions Human-derived TMPRSS2 protein is ev- olutionarilv conserved and functionally important.Hie results of this study can help to reveal the structure and mechanism of action of TMPRSS2 protein, provide ide¬as for the diagnosis and treatment of COYID-19, and accelerate the research and development process of new drugs targeting TMPRSS2 protein.

Treatment of cerebral palsy children by integrative medical sequential method: a clinical efficacy observation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the efficacy of integrative medical sequential method in treating cerebral palsy (CP) children's intelligence development, muscular tension, serum interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-alpha).</p><p><b>METHODS</b>Totally 111 CP children were randomly assigned to the control group (50 cases) and the treatment group (61 cases). All patients received comprehensive rehabilitation training and intravenous dripping of Monosialotetrahexosylganglioside Sodium Injection for 10 days. But those in the treatment group additionally received Chinese medical enema for brain resuscitation, relieving rigidity of muscles and activating collaterals for 14 days. Then they started another medication cycle and lasted for a total of 6 cycles. Serum IL-6 levels and TNF-alpha contents were determined before treatment. Scoring for muscular tension, Gesell score for intelligence development, contents of serum IL-6 and TNF-alpha were assessed before and after treatment in the two groups.</p><p><b>RESULTS</b>Compared with before treatment in this group, muscular tension, Gesell scores for intelligence development all decreased in the two groups (P < 0.05). As for inter-group comparison, the decrement was more obvious in the treatment group than in the control group (P < 0.05). The total effective rate was 86.9% in the treatment group and 76.0% in the control group (P < 0.05). The contents of IL-6 and TNF-alpha were obviously reduced in the treatment group and the control group after treatment (P < 0.01). The decrement was more obvious in the treatment group (P < 0.05).</p><p><b>CONCLUSION</b>The two treatment methods were effective for CP children, but the efficacy was superior in the treatment group than in the control group, indicating integrative medical methods could play a synergistic effect and optimize the treatment program for CP.</p>

Effect of astrocytes with different degrees of proliferation on multidrug resistance gene expression in rats with epilepsy / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the relationship of activated astrocytes and multidrug resistance gene (MDR) expression in rats with epilepsy.</p><p><b>METHODS</b>Astrocytes of neonatal Sprague-Dawley rats were separated and cultured. The cultured cells of passage 3 were activated by TNF-α for 2, 24 or 48 hrs. The culture media of cells with different degrees of proliferation were infused to the lateral cerebral ventricle of rats with epilepsy. The expression of MDR in the brain tissue was ascertained by PCR, immunocytochemistry and Western blot.</p><p><b>RESULTS</b>After 2 hrs of TNF-α stimulation, astrocytes began to proliferate, and reached a peak at 24 hrs. The expression of MDR in the brain tissue increased after infusion of culture medium of proliferated astrocytes in the TNF stimulation group compared with that in the control group without TNF stimulation. The level of MDR expression in the TNF stimulation group was positively correlated with the degrees of cell proliferation.</p><p><b>CONCLUSIONS</b>Proliferation of astrocytes can increase the expression of MDR in rats with epilepsy and is probably involved in the development of refractory epilepsy.</p>