Expression of antimicrobial peptide Cecropin P1 in Saccharomyces cerevisiae and its antibacterial and antiviral activity in vitro

BACKGROUND: Cecropin P1, acting as an antimicrobial, has a broad-spectrum antibacterial activity with some antiviral and antifungal properties. It is a promising natural alternative to antibiotics which is originally isolated from the pig intestinal parasitic nematode Ascaris suum. Many studies have shown that Cecropin P1 is helpful for the prevention or treatment of clinical diseases. Therefore, it is very necessary to establish a safe, nontoxic, and efficient expression method of Cecropin P1. RESULTS: The results indicated that the recombinant protein was about 5.5 kDa showed by Tricine­SDS­ PAGE and Western blot. And Cecropin P1 was efficiently secreted and expressed after 12 h of induction, with an increasing yield over the course of the induction. Its maximum concentration was 7.83 mg/L after concentration and purification. In addition, in vitro experiments demonstrated that Cecropin P1 not only exerted a strong inhibitory effect on Escherichia coli, Salmonella sp., Shigella sp., and Pasteurella sp., but also displayed an antiviral activity against PRRSV NADC30-Like strain. CONCLUSIONS: Collectively, the strategy of expressing Cecropin P1 in Saccharomyces cerevisiae is harmless, efficient, and safe for cells. In addition, the expressed Cecropin P1 has antiviral and antibacterial properties concurrently.

Research Progress of Thalidomide and Its Derivatives in Treatment of Myelodysplastic Syndrome--Review / 中国实验血液学杂志

Thalidomide and its derivatives have been used in the treatment of myelodysplastic syndrome (MDS) because of their anti-angiogenic and immunomodulatory effects. In recent years, some studies have found that thalidomide and its derivatives not only showed significant efficacy in lower-risk MDS patients with del (5q), but also showed advantages in non-del (5q) MDS patients. In addition, the discovery of its molecular targets and new substrates makes it possible to develop a new generation of immunomodulatory drugs (IMiDs) and to design IMiDs-based proteolysis-targeting chimeras. In this review, the new progress in mechanism and clinical application of thalidomide and its derivatives were summarized briefly, so as to provide a more scientific, reasonable and effective scheme to the treatment of MDS.

Research progress on the effect of low concentration atropine on the prevention and control of myopia in children and adolescents among children and adolescents / 中国学校卫生

Abstract@#Myopia is a common refractive error in the process of development. With the high incidence and low age of myopia in children and adolescents, it has become a worldwide public health problem, seriously endangering physical and mental health. The prevention and control of myopia has a long way to go. Low concentration atropine, as an M type choline receptor inhibitor, has been proved by medical research to effectively delay the progress of myopia. This paper reviews the relevant studies at home and abroad in recent years. The optimal concentration of low concentration atropine, the best clinical regimen and possible potential side effects were discussed. From the point of view of school public health, this paper discusses the significance of low concentration atropine for the prevention and control of myopia among school age students, in order to provide a reliable basis and new ideas for the follow up clinical application of atropine and myopia prevention and control strategies for school students.

Development of a new protocol for freeze-drying preservation of Pseudoalteromonas nigrifaciens and its protective effect on other marine bacteria

Background: Freeze-drying is known as one of the best methods to preserve bacterial strains. Protectant is the key factor affecting the survival rate of freeze-dried strains. In addition, salinity, bacterial suspension concentration, drying time, and other factors can also affect the survival rate of strains to varying degrees. At present, there are relatively few studies on freeze-drying preservation of marine bacteria. In the present study, we performed the freeze-drying protectant screening and optimized the preservation conditions for Pseudoalteromonas nigrifaciens, which is widely distributed in marine environment. The protective effects of the screened protectants were verified by 18 other marine bacterial strains. Results: The results indicated that the combination of 5.0% (w/v) lactose, 5.0% (w/v) mannitol, 5.0% (w/v) trehalose, 10.0% (w/v) skim milk powder, 0.5% (w/v) ascorbic acid and 0.5% (w/v) gelatin was the best choice for the preservation of P. nigrifaciens. The suggested salinity and concentration of initial cell suspension were 10 g/L NaCl and 1.0 × 109 CFU/mL, respectively. Furthermore, stationary-phase cells were the best choice for the freeze-drying process. The highest survival rate of P. nigrifaciens reached 52.8% when using 5­10% (w/v) skim milk as rehydration medium. Moreover, the other 18 marine strains belonging to Pseudoalteromonas, Vibrio, Photobacterium, Planomicrobium, Edwardsiella, Enterococcus, Bacillus, and Saccharomyces were freezedried under the abovementioned conditions. Their survival rates were 2.3­95.1%. Conclusion: Collectively, our results supported that the protectant mixture and parameters were beneficial for lyophilization of marine bacteria

Study on commodity specification and grade standard and quality evaluation of Galla Chinensis / 中国中药杂志

This paper is aimed to study the commodity specification and grade standard of Galla Chinensis,for standardizing market order and guide the market circulation,and provide a basis for standardization of Galla Chinensis. With 33 samples of Galla Chinensis from market as the object of the research,the herbal textual research and market research were carried out. Then the grading indicator were selected by the descriptive statistics,principal component analysis and cluster analysis,combining with production practice,the commodity specification and grade standard of Galla Chinensis were set out. The data of moisture,ash,gallic acid as the internal index were used to analyze the relationship between the quality difference between grades and the appearance characters and the intrinsic composition. Herbal textual research and market research found that the high quality of Galla Chinensis characterized with large,complete,wall thick,grayish brown characteristics,and Galla Chinensis could be divided into gallnuts and horned gall. Through principal component analysis and cluster analysis,combining actual production and herbal record,screening,the length,diameter,single weight,the number of 500 g were determined as 4 grading indicators,the commodity specification was divided into two: gallnuts and horned gall,the grades was divided into two: selected goods and mixed goods. The result of correlation analysis showed there was no significant correlation between the internal index and the appearance characters of Galla Chinensis. The result of multiple comparison showed that the ash content of the selected goods was smaller than that of the mixed goods,but it did not reach significant. The content of gallic acid of the selected goods and the mixed goods of gallnuts were higher than selected goods of horned gall,and higher than mixed goods of horned galls. Using the length,diameter,single weight,the number of 500 g as the appearance characters index. Combining with the herbal textual research and market research,we have divided two specifications and two grades for the commodity specification and grade standard of Galla Chinensis,which can provide a basis for industry standards and national standards.

Study on commodity specification and grade standard and quality evaluation of Dipsaci Radix / 中国中药杂志

Through the records of herbal and market investigations,the samples collected from different market and origin were analyzed,and the appearance character indexes were determined in order to revise the commodity specification and grade standard of Dipsaci Radix combined with production practice. This is also to analyze association of the appearance with quality different of intrinsic components. The investigation results indicated the root's long and thick was better,and atrovirens of fracture surface,there are most of the market is dominated by the ungraded goods. Through principal component analysis,variance analysis and cluster analysis,combining with the feasibility of actual operation and herbal record,the length,middle diameter and phloem color were filtrated and the specification was divided into 2 ranks: the selection and ungraded goods,and the grade was divided into 2 ranks for the selection goods: big,small selected goods. Moreover,there were no significant correlation between the appearance and the intrinsic quality index of Dipsaci Radix,the content of extractum were significantly positive correlation with the moisture,but had extremely negative correlation with the total ash. Multiple comparisons indicated that the content of the moisture,extractum and asperosaponin VI of the selected goods were higher than the ungraded goods,while the total ash content were lower,and they hasn't significant difference. The commodity specification and grade standard of Dipsaci Radix as a basis provide commodity specification and grade standard of communities and standardizing market trade order.

Application value of nerve root sedimentation sign in lumbar spinal stenosis revealed on MRI / 中国组织工程研究

BACKGROUND: Clinical value of the nerve root sedimentation sign in patients with lumbar spinal stenosis (LSS) is still uncertain.OBJECTIVE: To investigate the positive rate and correlation of the nerve root sedimentation sign in patients with LSS.METHODS: Totally 281 patients complaining lumbago or low back pain underwent MRI examination in the Orthopedic Clinic or Ward of Jinshan Hospital of Fudan University, including 119 males and 162 females. The cross-sectional area and sagittal diameter of the spinal canal were measured, and the patients were then divided into two groups according to the presence and absence of LSS. Finally, the positive rate of nerve root sedimentation sign was compared.RESULTS AND CONCLUSION: (1) Grouping based on the presence and absence dural sac cross-sectional area stenosis: the positive rate of nerve root sedimentation sign in sever, non, and mild LSS patients was 91.4%, 39.6%,and 53.5%, respectively, and the difference was significant (P < 0.05), but the positive rate showed no significant difference between mild and moderate as well as moderate and severe LSS patients. (2) Grouping according to the presence and absence spinal canal sagittal diameter stenosis: there was a significant difference in the positive rate of nerve root sedimentation sign between non-LSS and LSS patients (44.3% vs. 76.1%, χ2=21.469, P=0.000). (3) Grouping based on the dural sac cross-sectional area combined with spinal canal sagittal diameter: the dural sac cross-sectional area < 120 mm2 or the spinal canal sagittal diameter < 10 mm indicating LSS, there was a significant difference in the positive rate of nerve root sedimentation sign between non-LSS and LSS patients (40.6% vs.80.6%,χ2=30.539, P=0.000). (4) These results indicate that the diagnosis value of nerve root sedimentation sign in LSS is still under discussion. Although a positive sedimentation sign exclusively and reliably occurs in patients with severe LSS, it is also higher in patients without LSS. It may be inappropriate to take nerve root sedimentation sign as a diagnostic tool, but when combined with dural sac cross-sectional area and sagittal canal subsidence, it will be of great significance for the diagnosis of LSS.

Development of a Rapid and Sensitive Method for Detection of African Swine Fever Virus Using Loop-Mediated Isothermal Amplification

ABSTRACT A loop-mediated isothermal amplification (LAMP) assay was developed for rapid, sensitive and specific detection of African swine fever virus (ASFV). A set of LAMP primers was designed based on the sequence of the ASFV gene K205R. Reaction temperature and time were optimized to 64 oC and 60 min, respectively. LAMP products were detected by agarose gel electrophoresis or visually with the addition of fluorescent dye. The detection limit of the LAMP assay was approximately 6 copies of the target gene per microliter, 100 times more sensitive than conventional PCR. LAMP is a simple and inexpensive molecular assay format for ASFV detection. To date, African swine fever has not been reported in China. LAMP can be used to monitor ASFV spread into China, thereby reducing the threat of ASF.

Effect of bortezomib on migration and invasion in cervical carcinoma HeLa cell

OBJECTIVE@#To explore the effect of bortezomib on migration and invasion of cervical carcinoma HeLa cell and specific molecular mechanism.@*METHODS@#The effect of bortezomib on the viability of HeLa cell was measured by MTT assay. The effect of bortezomib on cell migration and invasion was measured by Transwell assay and invasion experiment respectively. The activation of Akt/mTOR signaling pathway and expression level of MMP2, MMP9 were assayed by western blot.@*RESULTS@#MTT assay indicated bortezomib (2.5 μM, 5 μM, 10 μM) could inhibit HeLa cell viability, and the inhibitory rate was highest at 48 h. Transwell assay and invasion experiment results showed that bortezomib inhibited HeLa cell migration and invasion. Western blotting assays presented bortezomib could suppress the phosphorylation of Akt and mTOR, and down-regulate the expression of MMP2 and MMP9.@*CONCLUSIONS@#These results suggested bortezomib could inhibit migration and invasion in cervical carcinoma HeLa cell, which might be related to Akt/mTOR signal pathway.

Benefit and Safety of Dual Antiplatelet Therapy after Coronary Artery Bypass Grafting for Off-pump CABG: A Systematic Review and Meta-analysis.

Aims: This study aims to evaluate benefit and safety compared dual antiplatelet therapy with single aspirin therapy after coronary artery bypass grafting. Study Design: A systematic review and Meta-analysis. Place and Duration of Study: Medline, Embase, ScienceDirect and Cochrane Library databases were searched for randomized controlled trials or observational studies focusing on anticoagulant therapy after coronary artery bypass grafting until December 2014. Methodology: Endpoints included postoperative mortality, bleeding events, myocardial infraction, stroke, repeat revascularization and graft occlusion. All these endpoints were compared between dual antiplatelet therapy and single aspirin therapy. Newcastle-Ottawa and Jadal scale were used to assess the quality of observational studies and randomized controlled trials respectively. Software R2.15.2 was utilized for Meta-analysis. Results: 15 studies composed of 31,365 patients were included. Compared with single aspirin therapy, dual antiplatelet therapy resulted in reducing risk of vein graft occlusion (OR=0.53, 95%CI 0.36-0.81, P=0.001), but no significant difference for artery graft occlusion (OR=0.91, 95%CI 0.39-2.12, P=0.882), Risk of postoperative mortality (OR=0.57, 95%CI 0.38-0.85, P=0.006) and repeat revascularization (OR=0.15, 95%CI 0.05-0.45, P=0.001) was also reduced. There were no significant difference for MI (OR=0.77, 95%CI 0.55-1.09, P=0.137), Stroke (OR=0.85, 95%CI 0.60-1.19, P=0.330) and bleeding (OR=0.95, 95%CI 0.82-1.09, P=0.465). In subgroup analysis of off-pump CABG, dual antiplatelet therapy reduced risk of graft occlusion (OR=0.49, 95%CI 0.30-0.82, P=0.006), MI (OR=0.28, 95%CI 0.11-0.72, P=0.009), mortality (OR=0.39, 95%CI 0.25-0.60, P<0.001), and did not increase risk of bleeding (OR=0.75, 95%CI 0.55-1.02, P=0.066). Conclusions: Dual antiplatelet therapy reduced risk of postoperative graft occlusion and mortality in the early and late postoperative phase after CABG. It appeared to be more beneficial for off-pump CABG.

Effect of bortezomib on migration and invasion in cervical carcinoma HeLa cell

Objective: To explore the effect of bortezomib on migration and invasion of cervical carcinoma HeLa cell and specific molecular mechanism. Methods: The effect of bortezomib on the viability of HeLa cell was measured by MTT assay. The effect of bortezomib on cell migration and invasion was measured by Transwell assay and invasion experiment respectively. The activation of Akt/mTOR signaling pathway and expression level of MMP2, MMP9 were assayed by western blot. Results: MTT assay indicated bortezomib (2.5μM, 5μM, 10μM) could inhibit HeLa cell viability, and the inhibitory rate was highest at 48h. Transwell assay and invasion experiment results showed that bortezomib inhibited HeLa cell migration and invasion. Western blotting assays presented bortezomib could suppress the phosphorylation of Akt and mTOR, and down-regulate the expression of MMP2 and MMP9. Conclusions: These results suggested bortezomib could inhibit migration and invasion in cervical carcinoma HeLa cell, which might be related to Akt/mTOR signal pathway.

Continuous renal replacement therapy in children with multiple organ dysfunction syndrome: A case series

There is a lack of definitive information regarding the precise indications, implementation, and outcomes of continuous renal replacement therapy (CRRT) for the treatment of critically ill children. Six children (three boys, three girls) aged from 3 days to 8 years, all of whom had multiple organ failure, were submitted to bedside CRRT using M60 filter membranes. Modified Port carbonate formula was used and clotting time was maintained between 20 and 30 minutes. Activated partial thromboplastin time was 1.5- to 2-fold normal. One patient discontinued treatment due to family decision. Marked improvements were seen in the remaining five patients, including normalization of blood urea nitrogen and creatinine levels, stabilization of electrolytes, and improvements in markers of organ function. Of note, one patient (a six-year-old male) underwent the treatment for 241 hours. All five patients were subsequently discharged and recovered uneventfully. CRRT is effective for the management of children who are critically ill due to multiple organ failure.

Development and evaluation of a loop-mediated isothermal amplification (LAMP) assay for rapid detection of Actinobacillus pleuropneumoniae based the dsbE-like gene

This paper reports on the development and validation of a loop-mediated isothermal amplification assay (LAMP) for the rapid and specific detection of Actinobacillus pleuropneumoniae (A. pleuropneumoniae). A set of six primers were designed derived from the dsbE-like gene of A.pleuropneumoniae and validate the assay using 9 A. pleuropneumoniae reference/field strains, 132 clinical isolates and 9 other pathogens. The results indicated that positive reactions were confirmed for all A. pleuropneumoniae strains and specimens by LAMP at 63ºC for 60 min and no cross-reactivity were observed from other non-A.pleuropneumoniae including Haemophilus parasuis, Escherichia coli, Pasteurella multocida, Bordetella bronchiseptica, Streptococcus suis, Salmonella enterica, Staphylococcus, porcine reproductive and respiratory syndrome virus (PRRSV), and Pseudorabies virus. The detection limit of the conventional PCR was 10² CFU per PCR test tube, while that of the LAMP was 5 copies per tube. Therefore, the sensitivity of LAMP was higher than that of PCR. Moreover, the LAMP assay provided a rapid yet simple test of A. pleuropneumoniae suitable for laboratory diagnosis and pen-side detection due to ease of operation and the requirement of only a regular water bath or heat block for the reaction.