Adult ; Comet Assay ; DNA Damage ; drug effects ; Female ; Humans ; Lipid Peroxidation ; drug effects ; Male ; Peracetic Acid ; toxicity

Objective To investigate the effect of RNA interference(RNAi) on inhibiting respiratory syncytial virus(RSV) replication through comparing the anti-RSV activities between pshRNA7816 and ribavirin in cell culture system.Methods The recombinated plasmid pshRNA7816 and ribavirin was added to HEp-2 cells.Methyl thiazolyl tetrazolium(MTT) assay was used to detect cytotoxicity of pshRNA7816 and ribavirin on normal HEp-2 cells and protective effects of them on RSV infected HEp-2 cells.The effects of pshRNA7816 and ribavirin on change of cytopathogenic effect(CPE) of HEp-2 cells induced by RSV infection were observed through microscopically.Results pshRNA7816 had not significant toxicity on the growth of HEp-2 cells,but the ribavirin had significant toxicity when the concentration above 1.0 mmol/L.The pshRNA7816 and ribavirin could alleviate the CPE of HEp-2 cells induced by RSV infection,but the pshRNA7816 showed a more potent inhibition than ribavirin.The inhibition rates of pshRNA7816 were significantly higher than the maximum inhibition rate of ribavirin on RSV infection(P

OBJECTIVE:To establish the quality standard for Tongmai pill. METHODS:TLC was used for the qualitative iden-tification of Salvia miltiorrhiza,Angelica sinensis and Carthamus tinctorius. HPLC was used for the content determination of feru-lic acid. The column was Diamonsil C18 with mobile phase of acetonitrile-0.1% aqueous phosphoric acid (17∶83,V/V) at a flow rate of 1 ml/min,the detection wavelength was 316 nm,the column temperature was 30 ℃,and the injection volume was 10 μl.. RESULTS:The TLC of S. miltiorrhiza,A. sinensis and C. tinctorius showed clear spots and good separation. The linear range of ferulic acid was 8-80 μg/ml(r=0.999 8);RSDs of precision,stability and reproducibility tests were lower than 2%;recovery was 97.2%-99.6%(RSD=1.2%,n=6). CONCLUSIONS:The standard can be used for the quality control of Tongmai pill.

Objective: To optimize the extraction technology of Chinese medicinal materials in Yanyanling dispersible tablets.Methods: Orthogonal design was used to study the effects of three factors, including the ratio of liquid to materials, extraction time and extraction times on the extraction rate of Chinese medicinal materials and the comprehensive score of gallic acid content and total solid yield was used as the index.Results: The optimum extraction conditions were as follows: the ratio of materials to liquid was 1∶10 (g·ml-1), the extraction time was one hour, and the extraction times was three.Conclusion: Under the optimum conditions, the extraction rate of gallic acid in Chinese medicinal materials in Yanyanling dispersible tablets is 0.058%, and the total solid yield is 21.4%.The optimal process is stable and feasible, which can provide reference for the production of Yanyanling dispersible tablets.

Objective To investigate the clinical effects and postoperative complications of arthrotomy and arthroscopy in repair of 170 menisci in 168 cases.Methods One hundred and sixty-eight patients with meniscus injury were repaired by arthrotomy or arthroscopy.They were 121 males and 47 females.There were 77 left knees and 91 right knees;117 medial menisci and 53 lateral ones.Their average age was 25.5?8.4 years old.Arthro- scopic repair methods included puncture and grinding,bio-absorbable meniscus arrow fixation,Outside-In suturing, Inside-Out suturing,Elite scuff instrument repairing,T-Fix fixation and FasT-Fix fixation techniques.The clinical results were assessed on the basis of symptoms,physical signs,Tegner scores and Lysholm scores of the cases. Postoperative complications were also investigated.Re-arthroscopic exploration was done for patients with obvious symptoms and physical signs.Results The average folluw-up time was 49.3?28.8 months.Their mean pre- operative Tegner score was 3.3?2.3,and their postoperative one 6.8?2.1 (P＜0.05).Their preoperative Lysholm score was 30.1?18.2,and their postoperative one 87.5?22.5 (P＜0.01).There were significant differences in Tegner and Lysholm scores before operation and after operation.Ninety-eight repaired menisci were rated as excellent(57.7%),57 as good (33.5%),10 as fair (5.9%),and five as poor (2.9%).The total ex- cellent and good result was 91.2%.Of the 19 patients with obvious symptoms and physical signs,re-arthroscopic exploration found no healing in five and partial healing in six.Postoperative complications included pain at the Outside-In suture nodes in three cases,referred pain at posterior articular capsule resulted from failed Outside-In meniscus anterior horn suturing in one case,and twinge at the meniscus arrow site in the posterior capsule in five cases.No serious lesion occurred at blood vessels or nerves.The postoperative complication incidence was 5.3%. Conclusion The eight methods of arthrotomy and arthroscopy to repair injured menisci investigated in our study can have a high successful rate and low perioperative and postoperative risk.

Using MS as basic medium, supplemented with 1.0 mg · L(-1) IBA, the adventitious roots of Tripterygium wilfordii were induced, and the good adventitious root culture system was established by leaves or callus induced by leaves as explants. The adventitious roots were also induced with 2.0-4.0 mg · L(-1) NAA and the good adventitious root culture system established by using suspension cells from callus as materials to induce adventitious root. The content of triptolide of three adventitious roots culture system were exceeded in the natural root bark. The content of triptolide of AR3 adventitious roots was the highest about 5.3 times as that in the natural root bark. By using 5 L stirred fermentor during pilot enlarge cultivation, compared with 250 mL flask cultivation, the adventitious roots increment and secondary metabolites content per liter medium showed no significant difference. The accomplishment of this analysis laid a foundation by tissue culture production of the secondary metabolites of T. wilfordii.
Culture Media ; chemistry ; metabolism ; Fermentation ; Plant Growth Regulators ; analysis ; metabolism ; Plant Roots ; growth & development ; metabolism ; Tissue Culture Techniques ; instrumentation ; methods ; Tripterygium ; growth & development ; metabolism

The coding sequence for the mature peptide of porcine lactoferrin (Plf) was synthesized according to the codon usage of lactobacillus, to establish optimized porcine lactoferrin Lactobacillus expression system. The gene was ligated into the Xho I/BamH I site of Lactobacillus expression vector pPG612.1 and the recombinant plasmid pPG612.1-plf was transformed individually into Lactobacillus casei ATCC393, Lactobacillus pentosus KLDS1.0413, Lactobacillus plantarum KLDS1.0344 or Lactobacillus paracasei KLDS1.0652 by electroporation. After induction with xylose, expression of the recombinant proteins was detected by Western blotting and confocal laser scanning microscopy. Secretion of recombinant Plf proteins from four recombinant species was determined quantitatively by ELISA. The antibacterial activities of recombinant proteins were measured by agar diffusion method. The result shows that Plf was correctly expressed in four species of recombinant lactobacillus, with molecular weight of about 73 kDa. The expression levels in recombinant Lactobacillus casei, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paracasei were 9.6 μg/mL, 10.8 μg/mL, 12.5 μg/mL and 9.9 μg/mL, respectively. Antimicrobial activity experiment shows that the recombinant proteins were active against E. coli, Staphylococcus aureus, Salmonella typhimurium, Listeria, Pasteurella. The recombinant Plf expressed by recombinant Lactobacillus plantarum showed the best antibacterial activity among all recombinant lactobacillus species. These data represent a basis for the development and application of porcine lactoferrin from recombinant lactobacillus.
Animals ; Anti-Bacterial Agents ; biosynthesis ; Lactobacillus ; metabolism ; Lactoferrin ; biosynthesis ; Recombinant Proteins ; biosynthesis ; Swine

To prepare ginseng saponin Compound K with ultrasound-assisted total zymolytic ginseng saponins. The conversion rate was taken as the index to detect the pre-treatment factors such as ultrasonic power and ultrasonic time, as well as the impact of enzymatic factors, such as pH value, temperature, concentration of substrate, dosage of enzyme and reaction time, on the conversion rate. The response surface method was used to optimize the preparation conditions. The enzymolytic products were identified with MS, 1H-NMR and 13C-NMR. The results showed that the optimum conditions of the ultrasound-assisted enzymolysis were 250 W for ultrasonic power, 15 min for ultrasonic time, 5.5 for enzymolytic pH, 50 degrees C for enzymolytic temperature, 36 h for enzymolytic time, 4:5 for enzymolytic dosage: substrate and 1.0 g x L(-1) for concentration of substrate. The relative molecular mass of reaction products was 622.4. Therefore, the nuclear magnetic map verified that the reaction product was rare ginseng saponin Compound K. Under the above conditions, based on the total zymolytic ginseng saponins, the conversion rate of rare ginseng saponin Compound K was 6.91% in proportion to the total of ginsenosides. The process features gentle reaction conditions, high conversion rate and simple and reliable process, which is suitable for industrial production.
Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Enzymes ; chemistry ; Panax ; chemistry ; Plant Roots ; chemistry ; Saponins ; chemistry ; isolation & purification ; Ultrasonics ; methods

Animals ; Autophagy ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the incidence and significance of perineural invasion (PNI) in cervical cancer. Methods Retrospective chart review of patients with cervical cancer (stages Ⅰa2-Ⅱb) who underwent radical hysterectomy and pelvic lymphadenectomy from 2007 to 2012. To evaluate the incidence and significance of PNI in cervical and uterine tissues by microscopic examination. Results A total of 238 patients were included, 9.2% (22/238) patients with PNI in the cervical stroma. Patients with PNI were more likely to have adverse histopathologic features, including lymphoma vascular space invasion, parametrical invasion, depth of invasion, tomor size and lymph nodes metastases (all P<0.05). PNI were independent of age, international federation of gynecology and obstetrics (FIGO) stage, histopathology type and grade, and positive vaginal margin (all P>0.05). Patients with PNI had shorter disease-free and overall survival (P=0.002 and P=0.008, respectively). On multivariate analysis, risk factors for recurrence and death included parametrical invasion and depth of invasion (P<0.05). Similarly, risk factors for recurrence included lymph nodes metastases (P=0.024). However, PNI was not identified as an independent risk factor for either recurrence or death (P>0.05). Conclusions PNI exists in early cervical cancer. PNI is associated with tumor size, depth of invasion, parametrical invasion, lymphoma vascular space invasion and lymph nodes metastases. PNI represente a decreasing disease-free and overall survival in patients with early-stage cervical cancer, and is independently associated with multiple high-risk factors, which be informed management decisions regarding adjuvant therapy.