In order to investigate the effect of malic enzyme on anaerobic mixed acid fermentation,the gene encoding NAD+-dependent malic enzyme was amplified from E.coli DH5? genome by using PCR and cloned into vector pTrc99a to give an expression vector pTrc99a-sfcA.Malic enzyme was overproduced by E.coli FMJ39(ldh,pfl)harboring pTrc99a-sfcA under the condition of IPTG induction.The results of anaerobic fermentation indicated overexpression of malic enzyme would have influence on the pathway of formate,acetate and succinate of FMJ39.The concentration of formate and acetate was 17.58% and 15.27% higher than FMJ39,the succinate concentration was reduced by 26.87%.There was no obvious citrate concentration change.The experimental results also indicated that high concentration of L-Thr and L-Ser would induce the Tdc operon to convert pyruvate into formate and acetate even when pfl gene encoding the pyruvate formate lyase was inactivated.

Objective: To study the effects of different origins, collection and processing methods on the quality of Taraxacum mongolicum. Methods:The HPLC fingerprints of Taraxacum mongolicum were established. Totally 11 batches of Taraxacum mongoli-cum were analyzed by similarity evaluation and cluster analysis. Results:According to the results of HPLC, 11 batches of Taraxacum mongolicum had good baseline separation and showed 5 common peaks. Based on the established HPLC method, the quality of different batches of Taraxacum mongolicum showed difference according to the results of similarity evaluation and cluster analysis. The quality of batch 121231-1 was the best. Conclusion:The origin, collection and processing method show notable influence on the quality of Ta-raxacum mongolicum, and the comprehensive score method can be applied in the quality evaluation of Chinese herbs.

OBJECTIVE:To determine the plasma mycophenolic acid concentration by HPLC, and study the multidoses pharmacokinetics character of mycophenolic acid in Chinese kidney transplantation patients. METHODS: The samples were precipitated with acetonitrile before injection. Diamonsil C18 column was used. The mobile phase consisted of acetonitrile-10 mmol?L-1 KH2PO4 (5∶6) at a flow rate of 1.1 mL?min-1. The detection wavelength was 254 nm and the column temperature was 40 ℃. This method was used to determine the multidoses pharmacokinetics in 12 kidney transplantation patients. RESULTS: MPA was well-separated from internal standard in chromatography, and endogenous foreign substance in plasma had no interference on the determination. The liner range for MPA was 0.38～59.00 ?g?mL-1,and the lowest detectable concentration of MPA was 0.38 ?g?mL-1. The recovery rate stood at 89.32%～97.63%; Both intra-day and inter-day RSD were less than 8.00%. Significant individual difference was noted among the patients treated with MMF in pharmacokinetic results, which was in line with the literature. CONCLUSION: This method is accurate and simple and applicable for the pharmacokinetics study of mycophenolic acid.

Adult ; Arsenic Poisoning ; etiology ; Arsenicals ; Humans ; Male ; Skin ; injuries ; Skin Absorption ; Wounds and Injuries

Objective To investigate the common cause,inducement and treatment response in patients with status epileptic(SE) in children,in order to improve the level of prevention and treatment of the disease.Methods Valium and chloral hydrate be used in the patient untreated,followed with luminal.Deparkin injection be used in the patient uncontrolled,followed with syrup.Results Of 21 patients,3 cases with epilepsy hadn′t been treated;6 cases with epilepsy hadn′t been treated regularlly;3 cases with epilepsy place respiratory infection;4 cases were the first attack with no inducement;3 cases were suffering from viral encephalitis.Four cases with epilepsy were died who hadn′t been treated regularly.Deparkin might be helpful in treatment with SE,but poor control in patients with long time attacks.Conclusions The attack and prognosis of SE are closely relative to original sick.It is important to prevent the attack.Early diagnosis is the key factor to improve prognosis.

Background Animal model of fungal keratitis is an available tool to the experimental study of the pathogenesis mechanism of fungal keratitis. Current modeling methods of fungal keratitis include corneal scratching, corneal stroma injection and corneal surface lens methods. But these methods still have their own shortages. Objective This experiment was to create a fungal keratitis animal model by modifying corneal surface lens method. Methods Modified animal models of fungal keratitis were created by modified corneal surface lens method in 12 general adult New Zealand white rabbits. The filter papers soaked 108 spores / ml or A106spores / ml of spergillus fumigatus suspension were attached on the de-epithelial cornea surface and fixed with contact lens and tarsorrhaphy for 2 days, and the filter paper with physiological saline was used as control group. The symptoms of anterior segment were examined under the slit lamp in 3 ,7 and 14 days after surgery and scored based on the criteria of Dong. Corneal scraping was stained with 10% potassium hydroxide and calcofluor white stain to observed mycelium under the fluorescence microscope. Corneal tissue sections were examined by hematoxylin-eosin staining and periodic acid Schiff staining under the light microscope. The use of animal followed the Standard of Association for Research in Vision and Ophthalmology. Results Fungal keratitis models were successfully established in 6 eyes and 4 eyes in 108 spores/ml group (6/6) and 106 spores/ml group respectively. The symptom was more severer and score was higher in the eyes of 108 spores/ml group than that in 106 spores/ml group. At 3 and 7 days after surgery,the symptom scores of fungal keratitis models were higher than those of control group from 3 through 7 days with the statistically significant difference (P<0. 01) and the symptom scores of 108 spores/ml group were significantly higher than those of 106 spores/ml group (P<0. 01). At 14 days after surgery, the symptom scores of 108 spores/ml group were still higher than those of control group (P<0. 05). Fungal hyphae was seen in the corneal scrapes in 108 spores/ml group and 106 spores/ml group respectively from 3 through 7 days after surgery. Inflammatory cell infiltration, stroma cells necrosis and fungal hyphae were presented in 108 spores/ml group, and the corneal neovascularization could be observed in 108 spores / ml group 14 days later. Fungal culture revealed the positive outcome in both 3 and 7 days after surgery in 108 spores/ml group,but in 106 spores/ml group,the positive result was only in the 3rd day. Conclusion Modified corneal surface lens method is more feasible and sample in the model of Aspergillus keratitis. This animal model of Aspergillus keratitis is practical for the further study of fungal keratitis.

This study was designed to prove simultaneously blocking both EGFR and COX-2-medi-ated pathways may be an efficient means of inhibiting cancer cell growth in NPC. Method: A combination of tarceva (EGFR-selectivetyrosine kinase inhibitors) with celecoxib(Cox-2 inhibitor) was studied on its effects on cell growth, cell cycle progression, apoptosis and protein expression in CNE-2 cell lines by cell growth assay, flow cy-tometric analysis assay and Western blotting. Result: ①The inhibition rate of cell growth was higher in the group treated with a combination of two agents than that the sum of rates of the two groups treated with only one agent (P<0.05). ②The combination of tarceva with celecoxib significantly induced G_1 arrest(P<0.05) ,but did not in-crease apoptosis rate(P>0.05). ③The group of combination showed less expressions of p-EGFR and COX-2 than any other group. Conclusion Simultaneously blocking EGFR and COX-2 mediated pathways would significantly in-hibit the growth of CNE-2 cell line, increase G_1 arrest and reduce the expression levels of p-EGFR and COX-2.

OBJECTIVETo compare the effect,safety,and advantage of flexible fixation with paperboard and pad versus short leg plaster in treating the fifth metatarsal base fracture,and establish the standard of diagnosis and treatment of the fifth metatarsal base fractures in flexible fixation with paperboard and pad.

METHODSFrom June 2010 to March 2013,59 patients with the fifth metatarsal base fracture were treated with paperboard and pad fixation or short leg plaster. Patients were enrolled and divided into paperboard and pad treatment group (paperboard group) and short leg plaster treatment group (plaster group) randomly according to the random number table. In paperboard group,there were 29 cases including 9 males and 20 females with an average age of (51.79±11.40) years old; the average course of injury was (11.59±6.58) hours. In plaster group, there were 30 cases including 9 males and 21 females with an average age of (52.13+17.34) years old ;the average course of injury was (11.03±7.06) hours. According to whether the fracture line across the articular surface, in paperboard group there were 14 cases of type A,15 of type B; in plaster group,16 of type A, 14 of type B. According to the degree of dislocation,in paperboard group there were 16 cases of degree I ,13 of degree II ; in plaster group,20 were degree I ,10 were degree II. Fracture was restored according to the type in manual. Patients in paperboard group were treated with paperboard and pad, and patients in plaster group were treated with short leg plaster. Fracture was fixed for 4 to 6 weeks according to fracture healing. On the 2nd, 4th,6th, 8th week and 3rd, 6th month after fixation, patients were followed up, and the foot function score was used to evaluate the function of injured foot. X-ray of injured foot was taken on the 2nd, 4th, 6th and 8th week were used to assess fracture healing.

RESULTSAll patients got complete follow-up. The X-ray result showed that all fracture reached at clinical healing on the 8th week after fixation without skin ulcer,nonunion and displacement of fracture. From the 4th to 8th week after fixation, paperboard group had a higher X-ray score than plaster group, but the difference between two groups had no statistically significance. Repeated analysis result showed that there was interact at different time point and between groups,the difference had statistically significance (P<0.01). The foot function score showed that at all time point, paperboard group had a higher score than plaster group, and on the 2nd, 4th, and 6th week, it had statistically significant difference(P<0.01) between two groups. On the 6th months after fixation,the excellent and good rate of paperboard group was 93.10%, higher than that of plaster group, which was 86.67%. But it had no statistically difference(P=0.483) between two groups.

CONCLUSIONUsing paperboard and pad fixation to treat the fifth metatarsal base fracture has the advantage of simplicity operating,reliable fixation, satisfactory effects, easily obtainable material.

Adult ; Aged ; Casts, Surgical ; Female ; Foot Injuries ; physiopathology ; surgery ; Fracture Fixation ; instrumentation ; methods ; Fracture Healing ; Fractures, Bone ; physiopathology ; surgery ; Humans ; Male ; Metatarsal Bones ; injuries ; physiopathology ; surgery ; Middle Aged

This study adopted a large sample, multicenter, registered hospital centralized monitoring nested prospective case-control study design. From the real world clinical application of Shenqi Fuzheng injection, monitored 30 026 cases of patients with the use of Shenqi Fuzheng injection. A total of 51 adverse drug reaction (ADR) cases was monitored, including 1 case of severe adverse reactions. ADR incidence rate was 1.7 per 1,000. Blood samples were collected from patients with allergic reactions and their matched controls. Related biological indicators of allergic reactions were unified detected and analysed in order to explore the mechanism of allergic reaction and promote the clinical safety.
Case-Control Studies ; Drug Hypersensitivity ; etiology ; Drugs, Chinese Herbal ; adverse effects ; Humans ; Injections ; Product Surveillance, Postmarketing ; Prospective Studies

matK is one of the core DNA barcode markers for plant DNA barcode identification and its universality using single makers has been in controversy. However, the universalities of different matK primer pairs in same seed plant group (order) and same matK primer pairs in different seed plant groups (order) are lack of systematic research. In this study, we collected 14563 full-length matK sequences of 11429 species of 3292 genera in 239 families belonging to 36 orders in seed plants. The universalities of 13 matK primer pairs and its 78 primer com-binations have been assessed using bioinformatics methods. The results indicated that xf/5r, 1F/8R, 390F/1326R and 3F_KIM/1R_KIM were the four most universality primer pairs. The four markers' universalities were 91.18%, 84.65%, 79.81% and 80.94% respectively in all 11429 seed plants. The most universality primer pairs in different orders were different. For each order, the primer pair with maximum universality was different. the xf/5r was the basal primer pair for primer combination and 1F/8R, 1F/1R, M3/M4 and 3F_KIM/1R_KIM could be the complementary primer pairs. This study could be a valuable resource for the primer selection of the research DNA barcoding identification in seed plants.