Wuxistatin, a novel and potent statin, is converted from lovastatin by Amycolatopsis sp. CGMCC1149. In the bioconversion, lovastatin is firstly hydroxylated by a hydroxylase. To obtain the critical hydroxylase, a novel hydroxylase gene was isolated from Amycolatopsis sp. CGMCC1149 by Degenerate PCR and Self-Formed Adaptor PCR and expressed in Escherichia coli. BLAST sequence analysis revealed that the gene belonged to cytochrome P450 gene superfamily and could encode a 403-amino-acid protein with a molecular weight of 44.8 kDa. The secondary structure prediction result showed that this protein contained many typical functional regions of P450, such as oxygen binding site, ion-pair region and heme binding region. Meanwhile, a catalytic function verification system was constructed by NADH, ferredoxin and ferredoxin reductase which could catalyze lovastatin hydroxylation into the target product. These would be helpful for further studies in large-scale production of wuxistatin.
Actinomycetales ; enzymology ; genetics ; Amino Acid Sequence ; Butyrates ; metabolism ; Cloning, Molecular ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Hydroxylation ; Industrial Microbiology ; Lovastatin ; metabolism ; Molecular Sequence Data

Objective To explore the effect of miR-126 on proliferation and apoptosis in colon cancer cells via targeting regulation of SOX2 expression. Methods miR-126 mimics and miR-126 NC were transfected into SW480 cells by liposome LipofectamineTM2000. The expression of miR-126 was detected by RT-PCR. Cell viability was determined by MTT staining. Cell apoptosis and cell cycle were detected by flow cytometry. The expression of SOX2 protein and mRNA was measured by western blot and RT-PCR. Luciferase reporter analysis was performed. Results Compared with miR-126 NC, the expression of miR-126 was upregulated(P< 0.01),cell viability was reduced(P< 0.01),early cell apoptosis rate and late apoptosis rate were increased(P< 0.01), cell cycle was arrested at G1 phase(P< 0.01), meanwhile, miR-126 mimics targeted downregulation of the expression of SOX2 protein and mRNA(P< 0.01). Conclusions miR-126 mimics can inhibit SW480 cell proliferation and induce cell apoptosis by targeting downregulation of expression of SOX2.

Objective To evaluate the safety of fibrin sealant (FS) intraperitoneal injection in SD rats .Methods 80 male and female SD rats were randomly divided into four groups (0 ,85.5 ,171 .0 ,342 .0 mg/kg) by body weight .All rats were in-traperitoneally injected with vehicle or FS daily for 14 days followed by a 28-day recovery period .The clinical signs ,hematolog-ical and biochemical indices were measured .The pathology were observed .Results Increase of white blood cell count (WBC) and decrease of fibrinogen (FIB) in d 14 were found in 171 .0 mg/kg and 342 .0 mg/kg dosage groups .Furthermore ,the tend-ency of weight increase of spleen were found in 171 .0 mg/kg and 342 .0 mg/kg dosage groups .Pathological exams of peritoneal cavity found that there were granulation tissues containing FS in some of the rats in 342 .0 mg/kg group .All of these changes got reversed after the recovery period .Conclusion The safety dose in this study is considered to be 85.5 mg/kg ,and the toxic-ity dose is 171 .0 mg/kg .The target toxicity systems or site of FS in SD rats are hematological system ,immune system and in-jection site .The toxic effects of FS are reversible .

Objective To explore the fluctuation characteristics of long-term doctor's order workload in pharmacy intravenous admixture services (PIVAS) and build a daily workload fluctuation prediction model and provide reference for the adjustment of PIVAS work mode. Methods Daily workload data of long-term doctor’s orders from PIVAS in the East Campus of Zhongshan Hospital affiliated to Fudan University from July 2020 to June 2021 were selected , and the time series analysis method was used to analyze the workload fluctuation characteristics and a prediction model was established. The accuracy of the model was verified by fitting parameters and prediction results. Results The fluctuation of PIVAS long-term doctor's daily workload data had the characteristics of periodicity, short-term slow rise and irregular variation. The Winters multiplier model was used to fit the series with R² = 0.777, the significance value of Ljung-Box statistic value (P value) was 0.060, and the mean absolute error percentage between the fitted and actual values was 4.45%, indicating that the model fitting accuracy was high. The average relative deviation between the predicted and actual results was 3.81%, indicating that the model prediction was effective. Conclusion The model constructed in this study could be used for the analysis and prediction of long-term doctor's orders workload of PIVAS. However, because the workload of doctor's orders has fluctuations such as periodicity and irregular changes, it is necessary to adjust the working model according to the fluctuation characteristics of the workload and the prediction results to ensure the efficient operation of PIVAS.

Vaccines are among the most effec-tive measures for preventing infectious diseases and play a crucial role in controlling the spread of these diseases.Adjuvants,serving as auxiliary com-ponents in vaccines,are indispensable in the vac-cine development process.Ideal adjuvants not only enhance the immune response,enabling the body to achieve optimal protective immunity but also play important roles in reducing the dosage of im-munogens and lowering vaccine production costs.To meet the demands of novel vaccines,many new types of adjuvants have been developed.However,there is still a lack of adjuvants that are safe,effec-tive,easy to prepare,highly pure,and suitable for a variety of vaccines in clinical settings.This article categorizes adjuvants and summarizes their mecha-nisms of action and characteristics,focusing on tra-ditional aluminum salt adjuvants and more modern lipid-based and nucleic acid-based adjuvants.The summary is based on a computer search of data-bases including PubMed,Embase,The Cochrane Li-brary,CNKI(China National Knowledge Infrastruc-ture),VIP Database,and Wanfang Database,using English search keywords such as Adjuvants,Vac-cine,Vaccine Adjuvant,aluminum salts,MF59,AS03,Toll-like receptor agonist,etc.,and corre-sponding Chinese search terms.The aim is to pro-vide references for the development and applica-tion of adjuvants.