An Enzymolysis-Assisted Agrobacterium tumefaciens-Mediated Transformation Method for the Yeast-Like Cells of Tremella fuciformis

Agrobacterium tumefaciens-mediated transformation (ATMT), as a simple and versatile method, achieves successful transformation in the yeast-like cells (YLCs) of Tremella fuciformis with lower efficiency. Establishment of a more efficient transformation system of YLCs is important for functional genomics research and biotechnological application. In this study, an enzymolysis-assisted ATMT method was developed. The degradation degree of YLCs depends on the concentration and digestion time of Lywallzyme. Lower concentration (≤0.1%) of Lywallzyme was capable of formation of limited wounds on the surface of YLCs and has less influence on their growth. In addition, there is no significant difference of YLCs growth among groups treated with 0.1% Lywallzyme for different time. The binary vector pGEH under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter was utilized to transform the enzymolytic wounded YLCs with different concentrations and digestion time. The results of PCR, Southern blot, quantitative real-time PCR (qRT-PCR) and fluorescence microscopy revealed that the T-DNA was integrated into the YLCs genome, suggesting an efficient enzymolysis-assisted ATMT method of YLCs was established. The highest transformation frequency reached 1200 transformants per 106 YLCs by 0.05% (w/v) Lywallzyme digestion for 15 min, and the transformants were genetically stable. Compared with the mechanical wounding methods, enzymolytic wounding is thought to be a tender, safer and more effective method.

A Rapid and Effective Colony PCR Procedure for Screening Transformants in Several Common Mushrooms

In the post-genomic era, gene function analysis has attracted much attention. Transformation is often needed to investigate gene function. In this study, an easy, rapid, reliable, and cost-effective colony polymerase chain reaction (PCR) method for screening mushroom transformants was developed: picking up a suitable amount of transformant's tissue (1–10 μg) to 20 μl 0.25% Lywallzyme solution, and vortexing for 10 s followed by incubation at 34 °C for 15 min. Finally, 2 μl of the suspension was used as templates to perform PCR and single target bands were successfully amplified from respective transformants of Tremella fuciformis, Pleurotus ostreatus, and Pleurotus tuber-regium. This procedure could be widely employed for screening transformants in mushroom transformation experiments.

An Improved Total RNA Extraction Method for White Jelly Mushroom Tremella fuciformis Rich in Polysaccharides

An improved method for extracting high quality and quantity RNA from a jelly mushroom and a dimorphic fungus—Tremella fuciformis which is especially rich in polysaccharides, is described. RNA was extracted from T. fuciformis mycelium M1332 and its parental monokaryotic yeast-like cells Y13 and Y32. The A260/280 and A260/230 ratios were both approximately 2, and the RNA integrity number was larger than 8.9. The yields of RNA were between 108 and 213 µg/g fresh wt. Downstream molecular applications including reverse transcriptional PCR and quantitative real-time PCR were also performed. This protocol is reliable and may be widely applicable for total RNA extraction from other jelly mushrooms or filamentous fungi rich in polysaccharides.

Medication Monitoring of Anti-infection Drugs in a Patient with Lupus Nephritis Complicated with Com-munity Acquired Pneumonia / 中国药师

Objective:To explore the value of clinical pharmacists in the anti-infection practice for a patient with lupus nephritis complicated with community-acquired pneumonia(severe). Methods:Clinical pharmacists carried out medication monitoring in the entire treatment process of a patient with lupus nephritis complicated with community-acquired pneumonia( severe)in the clinical front line. Depending on the pharmaceutical knowledge,clinical pharmacists assisted physicians to make out the optimal medication and ad-justed the medication according to the patient′s conditions. Results:Moxifloxacin and piperacillin sodium /tazobactam sodium could ef-fectively control the severe pulmonary infection of the patient with lupus nephritis and avoid the potential risk of recurrence or aggrava-tion of lupus nephritis. Conclusion:The choice of antimicrobial agents for the special populations should consider not only the antibac-terial spectrum and antibacterial activity,but also the individual characteristics of patients and pharmacokinetics of drugs in order to im-prove the safety and effectiveness of medication.

Effect of Pulse Microwave Combined with Ofloxoacin on Chronic Prostatitis / 中国康复理论与实践

@#Objective To observe the effect of pulse microwave combined with antibiotics on chronic prostatitis.Methods100 outpatients with chronic prostatitis were randomly divided into the trail group and control group with 50 cases in each group. The patients of the trail group were treated with pulse microwave combined with Ofloxoacin, and those of the control group were treated only with Ofloxoacin. The treatment course was 5 weeks. The evaluation with National Institutes of Health-Chronic Prostatitis Symptom Index (NIH-CPSI) and test of white blood cell (WBC) count in prostate secretion were performed before and after treatment for all patients of two groups.ResultsIn the trail group, the mean symptom degree score of NIH-CPSI was 9.3±2.6 and living quality score was 3.4±1.2, the total score was 12.4±4.1, and WBC count was 13.7±5.4/HP. In the control group, these indexes were 13.3±3.5, 4.5±1.2, 17.5±4.6 and 17.1±5.2 respectively. The NIH-CPSI score and WBC count of the trail group were lower than that of the control group ( P<0.05), and cure rate and total effective rate were higher than that of the control group ( P<0.05).ConclusionThe treatment of pulse microwave combined with Ofloxoacin for chronic prostatitis can improve the cure ratio and efficiency.

L-hydroxyproline changes in rats' skin induced by ultraviolet C irradiation / 中国康复理论与实践

@#ObjectiveTo investigate the L-hydroxyproline (L-Hyp) changes in rats' skin after exposed to different doses of ultraviolet C irradiation (UVC) and the effect of UVC radiation on collagen synthesis.MethodsAfter the animal model was set up, each male Wistar rat was made three fresh skin wounds, and three skin wounds of rats were divided into the 15 MED UVC irradiation (15mJ/cm2),60 MED UVC irradiation (60mJ/cm2) and control (without UVC irradiation). Then the chemistry method was utilized to research changes of L-Hyp contents of the granulation.ResultsFrom the 21st to the 28th day after UVC irradiation, contents of L-Hyp in the 15 MED group were higher than that in controls (P<0.05). While at the 28th day, L-Hyp level in the 60 MED group increased greatly and was higher than that in other two groups (P<0.01).ConclusionUVC irradiation increases L-Hyp level in rat's wound skin, so it accelerates the collagen synthesis and is helpful to promote wound healing, and the effect of 60 MED dose is superior to that of 15 MED.