Objective To investigate survivin as an anticancer therapeutic target by use of shepherdin [79-87],a novel peptide carrying the survivin sequence from Lys-79 through Leu-87,we constructed an recombinant vector containing fusion gene NT4-Ant-Shepherdin [79-87].Methods The gene of Ant-Shepherdin [79-87] was obtained by PCR and T-vector method.After cloned and digested with restricted enzyme,Ant-shepherdin [79-87] was inserted in PBV220NT4 vector.The recombinant vector was transformed into the competent cell,E.coli DH5?.The fusion gene of NT4-Ant-Shepherdin [79-87] was identified by agarose gel electrophoresis (AGE).Results DNA sequencing results verified that the sequence of Ant-Shepherdin [79-87] was consistent with what we had designed.After transformed E.coli DH5?,a fragment of 321 bp was confirmed.Conclusion The recombinant vector containing fusion gene NT4-Ant-Shepherdin [79-87] was successfully constructed in this experiment by molecular biology techniques,which provides the basis of further research of survivin for cancer gene therapy.

Objective To study the protective effect of pure carbachol or combined with dietary fiber on intestinal mucosal barrier of rats after diffuse brain injury (DBI).Methods An adult male Wistar rat model of DBI was reproduced by gravitational shock method. The rats injured and survived after resuscitation were divided into three groups: model group (n = 40), carbachol group (n = 40) and carbachol combined with dietary fiber group (combined group,n = 32). In addition, a control group was established by simply an incision performed on the scalp, and the rats could drink freely (n = 5). In the experimental groups, 2 hours after resuscitation the rats began to receive gavage, 6 hours once, the liquid amount 15 mL/kg should be assured in every 6 hours, and if insufficient, normal saline was supplemented. In model group, normal saline 90 mL/kg was given, in carbachol group, carbachol 300μg/kg was administered and in combined group, carbachol 300μg/kg combined with dietary fiber 60 mL/kg was supplied. At 3 (combined group being excluded), 6, 12, 24 and 48 hours after resuscitation, the rats were anesthetized to collect samples and detect the plasma levels of D-lactate and activity of diamine oxidase (DAO) respectively, and the changes of villus height of small intestine were examined by a light microscope.Results The plasma D-lactate levels and the activities of DAO at any time point in the experimental groups were significant higher than those in control group (allP < 0.01). Along with the prolongation of time, the levels of plasma D-lactate and DAO activities in carbachol and carbachol plus diatary fiber groups were gradually lower than those of the model group, and at 48 hours after injury they reached their valley values [D-lactate (ng/L): 6.32±0.79, 7.46±1.67 vs. 17.65±1.53, DAO activity (kU/L): 0.76±0.01, 0.86±0.01 vs. 2.23±0.15]. Under light microscopy, compared with control group, the villus height of small intestinal mucosa at any time point in any experimental group was gradually lowered, and reached the valley values at 12 hours, then gradually increased , and peaked at 48 hours, the villus height in carbachol group and combined group was higher than that in model group (μm: 265.36±10.20, 261.54±10.38 vs. 247.51±9.39, bothP < 0.05).Conclusion When only carbachol is administered into the rat intestine early after diffuse brain injury in rats, beginning from 6 hours after injury, the protective effect of intestinal mucosal barrier is shown, representing decrease of plasma D-lactate level and DAO activity, amelioration of intestinal mucosal damage and protection of intestinal mucosal barrier; under the same above situation, the carbachol combined with dietary fiber was applied, showing the similar above carbachol protective effects.

Objective To investigate the distribution of occupations and traditional Chinese medical syndromes of the patients with hypokalemic periodic paralysis in Foshan area. Methods A multicenter retrospective investigation was carried out to analyze the characteristics of occupations and syndrome types of hypokalemic periodic paralysis in Foshan area, and the correlation of occupations with syndrome types was also explored. Results (1) The workers engaged in agriculture, forestry, animal-breeding and fishing, and the workers engaged in production & transportation were most likely to suffering from hypokalemic periodic paralysis, with the incidence being 36.8%, 34.7% respectively. (2) The damp-heat syndrome was the most commonly-seen syndrome type, accounting for 53.1%, and then followed by Qi deficiency syndrome (20.3%) and Qi-Yin deficiency syndrome(15.7%).(3) The syndromes of heavy manual labor workers such as agriculture, forestry, animal-breeding and fishing workers, production & transportation workers, and soldiers were characterized by damp-heat type, accounting for 62.5%, 69.4%, 47.0% respectively. Professionals & technicians were most likely to suffering from Qi-Yin deficiency syndrome, accounting for 44.4%; business service personnel were most likely to suffering from Qi-Yin deficiency syndrome (32.5%) and Qi deficiency syndrome (31.3%). The syndrome distribution of heavy manual labor workers differed from that of light manual labor workers and brain workers(P < 0.001). Conclusion The high-risk groups of hypokalemic periodic paralysis in Foshan area are the heavy manual labor workers who are manifested with the damp-heat syndrome. The dominated syndrome types of light manual labor workers and brain workers are Qi deficiency and Qi-Yin deficiency.

Objective To analyse the effect of specimen storage time on platelet count in apheresis Donors,in order to choose the appropriate testing time.Methods We choose fifty healthy Unpaid Blood Donors of Platelets to test platelet count in 0,0.5,1,3,6h respectively by the blood counting instrument.Results At room temperature,the count of platelets from blood samples is relatively lower at 0 hour and the difference is significant (P ＜ 0.05).After 0.5-6 hours,the count of platelets become stabilized and not has significantly different(P＞0.05).Conclusion The count of platelets of blood samples is lower at 0 hour than 0.5-6 hours,this work suggest that the count of platelets of blood samples should be done at 0.5-6 h in order to protect platelets quality.

OBJECTIVE:To study the effect of ethanol on the pharmacokinetics of phenytoin sodium in rabbits METHODS:The serum concentrations of phenytoin sodium at different points of time were determined by UV-spectrophotometry in eight rabbits after administration of phenytoin sodium(10mg/kg)alone and in combination with ethanol RESULTS:After administration in combination with ethanol,the AUC of phenytoin sodium was significantly decreased from(4 108 64?1 039 98)mg/(min L) to (1 903 65?1 003 40)mg/(min L),Cmax from(29 0?2 94)mg/L to(16 0?5 9)mg/L,T1/2(ke) from(98 45?26 4)min to(82 84?25 5)min;but the apparent distribution volume(Vd)was obviously increased from(0 3 475?0 0 360)L/kg to(0 6 819?0 1 901)L/kg and the clearance rate(CL) from(0 0 026?0 0 008)ml/(kg?min)to(0 0 062?0 0 022)ml/(kg?min) CONCLUSION:The elimination of phenytoin sodium was significantly accelerated after simultaneous administration of ethanol in rabbits

Objective To observe the protection of carbachol on intestinal barrier function in patients with trauma. Methods A prospective randomized controlled trial was conducted. Seventy patients after trauma with a definite diagnosis of multiple organ dysfunction syndrome(MODS)from Department of Critical Care Medicine in Hebei United University Affiliated Hospital were included. According to random number table,the patients were divided into a carbachol treatment group(37 cases)and a mosapride citrate treatment group(33 cases),and all the patients in the two groups were treated by antibacterial drugs,supportive agents for organ function,surgery, etc symptomatic treatment. Based on the conventional treatment,in the carbachol treatment group,carbachol was administered through a stomach tube at the dose of 0.2 mg/kg,twice a day,and the dose was doubled if no exhaust or defecation persisted for 3 days after treatment,while in the mosapride group,mosapride citrate was given at the dose of 5 mg once and thrice a day,the therapeutic course of both groups being 7 days. On the 1st,3rd,5th, 7th day after admission,peripheral venous fasting blood in early morning was collected,the activity of diamine oxidase(DAO),expression rates of CD11b+and CD18+in polymorphonuclear neutrophil(PMN),contents of tumour necrosis factor-α(TNF-α)and interleukin-10(IL-10) were detected,and the clinical curative effects were observed. Results Compared to the mosapride citrate treatment group,the total effective rate was significantly higher in the carbachol treatment group on the 7th day after treatment〔70.3%(26/37)vs. 45.5%(15/33),P<0.05〕. The activity of DAO,expression rates of CD11b+and CD18+in PMN,contents of TNF-αand IL-10 in the carbachol treatment group were decreased with the extension of time,and reached valley values on the 7th day,the differences were statistically significant in the comparisons with those in mosapride citrate treatment group at the same time point〔DAO(mg/L):3.21±0.52 vs. 3.93±0.51,CD11b+:(14.89±2.16)% vs.(28.92±1.59)%,CD18+:(53.67±2.44)% vs. (72.46±4.08)%, TNF-α(ng/L):111.44±16.42 vs. 129.73±18.74, IL-10(ng/L):67.71±38.83 vs. 121.45±40.23,all P<0.05〕. At the various time points,the above indexes had no obvious changes in mosapride citrate treatment group. Conclusion Carbachol can ameliorate the ischemic/reperfusion(I/R)injury in patients with intestinal barrier dysfunction after trauma,decrease the release of inflammatory cytokines in vivo,and promote peristalsis of intestinal tract,therefore carbachol has clinical value of protecting intestinal barrier function.

OBJECTlVE To develop an LC-MS/MS method for simultaneous determination of pro-damine ( PDM) and its metabolite 2,4-dinitro-N3-propyl-6-trifluoromethyl-1,3-benzenediamine ( DTB) in rat plasma in order to study toxicokinetics of PDM in rats. METHODS SD male rats were administered a single dose of PDM ( ig: 100 and 1000 mg·kg-1; iv: 100 mg·kg-1 ) . LC-MS/MS method was used to determine PDM and DTB in rat plasma. Toxicokinetic parameters were fitted using DAS Ver2. 1. 1. RESULTS After ig administration of PDM 100 mg·kg-1 , the parameters of PDM and DTB were as fol-lows:AUC(0-t) was 2715±102 and (6845±316)μg·h·L-1, t1/2z was 9.0±1.4 and (7.1±1.3)h, Tmax was 7.0± 1.6 and (7.0±0.0)h, cmax was 146±51 and (473±103)μg·L-1. After ig administration of PDM 1000 mg·kg-1, the parameters of PDM and DTB were as follows:AUC(0-t) was 3401±242 and (10364± 573)μg·h·L-1, t1/2z was 8.8±2.1 and (6.0±1.8)h, Tmax was (7.0±1.6)h, cmax was 175±56 and (586± 152)μg·L-1 . The absolute bioavailability of PDM was 44.9%( 100 mg·kg-1 ) and 17.1%( 1000 mg·kg-1 ) . CONCLUSlON This method is suitable for the analysis of PDM and DTB in rat plasma. There is evidence that PDM and DTB display nonlinear toxicokinetic characteristics in the studied dose range.

0.05).Compared with control group,the GK activity of liver cell,the expression of PEPCK and the expression of GLUT4 in model group decreased signifi cantly(P

Objective:To determine the half lethal dose(LD50) of paraquat in rats and to establish a relatively safe and stable pathological animals model of pulmonary fibrosis.Methods: Ninety-six SD rats totally in half genders.Fifty SD rats in half genders were randomly divided into 5 groups,each had 10 rats.Feed the rats with different doses of liquor of paraquat intraperitoneally one time and definite the half lethal dose of one and two weeks.After that,prepare another forty-six SD rats,also in half genders,as intonication group,twenty-eight rats were treated with the liquor of paraquat in dosage of 18 mg/kg intraperitoneally.As control group,sixteen rats were treated with equivalent volume of normal saline.Observe the toxic symptom daily and rats were sacrificed on day 1,3,5,7,14,21,28,35 and 42 respectively for the histological examination.Results: The half lethal doses of intraperitoneal paraquat of 1 and 2 weeks were 18.27 and 17.29,with 95% confidence intervals of 16.61-20.09 and 15.99-18.67,respectively.After intraperitoneal paraquat injection at the dose of 18 mg/kg,typical toxic symptoms were observed at different times in the rats.The whole process of acute lung injury and fibrosis induced by paraquat intoxication could be seen with the naked eyes or under the light microscope.Conclusion: Paraquat has a strong toxicity to rats.A proper dose of paraquat solution can not only reduce the number of experimental rats,but also induce typical pulmonary fibrosis in rats.

Objective To study the effects on electroporation of Qingfengteng cataplasma transdermal absorption and describe the characteristics of animal pharmacokinetics of it.Methods Two-chamber diffusion cell was used and the plasma drug concentration was determined by HPLC.The application of AIC theory to analysis of the compartmentally based model of sinomenine transdermal delivery by electroporation.Results The Cmax,Ka,and AUC0→∞ of electroporation was larger than those of passive diffusion;t1/2(Ka)and tmax of electroporation were reduced compared with passive diffusion.The drug concentration-curve equation were C=2.884?(e-0.056 t-e-0.232 t)and C=2.512?(e-0.058 t-e-0.149 t)for electroporation and passive diffusion,respectively.Conclusion The change of in vivo drug concentration of Qingfengteng calaplasma transdermal absorption by electroporation could be analized in accordance with mammillary one-compartment open model.The etrectroporation technology could sharply enhance the bioavalibility compared with the passive diffusion.