Objective To assemble fusion gene of c-myc tagged human trefoil factor family 2(hTFF2)(in vitro) and construct a cloning vector of this fusion gene.Methods Based on amino acid sequence of hTFF2 and codon of lactococcus lactis,cDNA of htff2 sequence was designed and extended at their 5' ends with a sequence encoding the c-myc tag;and the sequence of fusion gene c-myc-htff2 was designed.According to restriction enzyme sites of pBluescript Ⅱ sk(+),the SalⅠ and BamHⅠ were arranged at 5′ and 3′ ends of the fusion gene respectively.Instructed by DNAWORKS program,the fusion gene sequence of c-myc-htff2 was designed as 14 oligonucleotides that overlapped each other.The target gene fragment of cmyc-htff2 fusion gene was obtained by the means of polymerase chain reaction(PCR) based gene assembly.Then,the c-myc-htff2 was subcloned into vector of pBluescript II sk(+) for identification of the fusion gene by restricting enzyme excision and DNA sequencing.Results Assembly of c-myc-TFF2 fusion gene(in vitro) and construction of pBS-TFF2 had been completed successfully;and DNA sequencing showed that the sequence of synthetic gene accorded with the expectation.Conclusion With the help of DNAWORKS program,the design and assembly of oligonucleotides (in vitro) is effective to synthesize a target gene,and the cloning vector of c-myc-htff2 fusion gene has been successfully constructed.

Mesaconitine was incubated with rat liver microsomes in vitro. The metabolites of mesaconitine in rat liver microsomes were identified by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method with high resolution power. A typical reaction mixture of 100 mol L-1 Tris-HCI buffer (pH 7.4) containing 0.5 gL-1 microsomal protein and 50 micro molL-1 mesaconitine was prepared. The above reaction mixture was divided into six groups, and the volume of each group was 200 micro L. The incubation mixture was pre-incubated at 37 degrees C for 2 min and the reactions were initiated by adding NADPH generating system. After 90 min incubation at 37 degrees C, 200 micro L of acetonitrile was added to each group to stop the reaction. The metabolites of mesaconitine were investigated by UPLC-MS/MS method. Mesaconitine and 6 metabolites M1-M6 were found in the incubation system. The structures were characterized according to the data from MS/MS spectra and literatures. The metabolic reactions of mesaconitine in rat liver microsomes included the demethylation, deacetylation, dehydrogenation and hydroxylation. The major metabolic pathways of mesaconitine in rat liver microsomes were determined by UPLC-MS/MS on multiple reaction monitoring (MRM) mode combined with specific inhibitors of cytochrome P450 (CYP) isoforms, including alpha-naphthoflavone (CYP1A2), quinine (CYP2D), diethyldithiocarbamate (CYP2E1), ketoconazole (CYP3A) and sulfaphenazole (CYP2C), separately. Mesaconitine was mainly metabolized by CYP3A. CYP2C and CYP2D were also more important CYP isoforms for the metabolism reactions of mesaconitine, but CYP1A2 and CYP2E1 haven't any contribution to MA metabolism in rat liver microsomes.
Aconitine ; analogs & derivatives ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 CYP3A Inhibitors ; Cytochrome P-450 Enzyme Inhibitors ; Cytochrome P-450 Enzyme System ; metabolism ; Enzyme Inhibitors ; pharmacology ; Ketoconazole ; pharmacology ; Male ; Metabolic Networks and Pathways ; Microsomes, Liver ; enzymology ; metabolism ; Quinine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sulfaphenazole ; pharmacology ; Tandem Mass Spectrometry

Firstly discovered in 1980s,human immunodeficiency virus (HIV) continues to affect more and more people.However,there is no effective drug available for the therapy of HIV infection.Betulinic acid existing in various medicinal herbs and fruits exhibits multiple biological effects,especially its outstanding anti-HIV activity,which has drawn the attentions of many pharmacists.Among the derivatives of betulinic acid,some compounds exhibited inhibitory activities at the nanomolar concentration,and have entered phase Ⅱ clinical trials.This paper summarizes the current investigations on the anti-HIV activity of betulinic acid analogues,and provides valuable data for subsequent researches.

OBJECTIVETo evaluate the effects of Qingre Liqi Granule (QLG) on clinical therapeutic efficacy, electrogastrogram (EGG) and gastric emptying in patients with functional dyspepsia (FD).

METHODSThirty-two FD patients of dyskinesis type enrolled were treated with QLG by oral taking for 6 days. Scoring on 8 kinds of symptoms, including abdominal distension, abdominal pain, morning gastric fullness, belching, regurgitation, nausea, vomiting, and anorexia, fasting EGG and the gastric emptying determination were performed using single photon emission computed tomography (SPECT) before and after treatment.

RESULTSThe total and individual scores of clinical symptoms, expect that of vomiting, significantly decreased after treatment (P < 0.05), and the percentage of patients with tachygastria and bradygastria significantly decreased (P<0.01) at the same time. EGG after treatment showed significantly elevated rates of normal slow wave dominant power, and nearly normalized dominant frequency. An increased gastric emptying rate at different phases after 75 min (P < 0.05), and significantly shortened gastric emptying half-time (P < 0.01) were shown meanwhile in gastric emptying detection. The improvement of symptom score and gastric emptying half-time showed significant positive linear correlation (r =0.8929, P < 0.01).

CONCLUSIONQLG can improve symptoms of FD patients by regulating the rhythm and power of gastric electro-wave, increasing gastric motility and enhancing gastric emptying function.

Adult ; Aged ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Dyspepsia ; drug therapy ; Electromyography ; drug effects ; Female ; Gastric Emptying ; drug effects ; Humans ; Male ; Middle Aged ; Myoelectric Complex, Migrating ; drug effects ; Phytotherapy ; Treatment Outcome ; Young Adult

Objective:To study the prosthodontic design for the treatment of maxillary first molar mesio-lingual cusp defect with subfis-sue.Methods:A finite element model of maxillary first model,including mesio-lingual cusp defect with subfissue,periodontal support-ing tissue and a section of the maxilla was established by cone beam CT.Different prosthodontic designs for respective restoration and simulating different bite force were adopted,the Von Mises stress,maximum compressive stress and the crack distribution,J integral and the root displacement were analysed.Results:In the metal pile restoration model,the stress,JINT value and the equivalent stress and displacement of root were the smallest.With the increase of crack depth of the vertical and oblique fissue,the equivalent levels of stress (equivalent stress of J integral),the stress experienced by the dentin along with the peak of the maximum principal,increased signifi-cantly.Under 4 kinds of force direction and 3 repair methods,the equivalent stress peak value,J integral dentin stress,maximum prin-cipal stress peak value and dentin stress peak value of the cracks was 200 N,90°>200 N,45°>600 N,0°>200 N.Conclusion:High elastic modulus and all ceramic crown may lead to minimal equivalent stress,J integral,dentin equivalent stress and maximum principal stress.When subfissure is repaired,teeth fracture remains possible and any possible propagation may exist.The effect of later-al force on the crack and the dentin is greater than that of vertical force for crack expansion and tooth fracture.

ObjectiveTo investigate characteristics in vitro and contrast-enhanced effect in vivo with self-made nano-scale ultrasoud-enhanced contrast agent.MethodsNano-lipid microbubbles were prepared by machine vibration and low-speed centrifugation.The surface potential and the microbubbles size were assessed by Zetasizer nano ZS90.The bubbles dispersity and micromorphous were observed under light microscopy and electron microscope.The characteristics of microbubbles were studied at various times at 4℃ environment.The enhancement duration time and intensity of nano-lipid microbubbles in nude mice livers,kidneys and subcutaneous implanted tumors were observed,and compared with micro-scale microbubbles.ResnltsThe mean diameter of nano-lipid microbubbles was (580.6 ± 36.3) nm.The contrast imaging study in vivo showed the nano-lipid microbubbles could significantly enhance ultrasonic imaging in nude mice livers,kidneys and subcutaneous implanted tumors.ConclusionsSelf-made nano-scale ultrasoudenhanced contrast agent showed good stability and contrast-enhancement effect in vitro,and provide a experimental basis for ultrasound imaging and therapeutics of malignant tumors.

Objective To Study the effects of high mechanical index ultrasound radiation microbubble on colon cancer cell skeletal.Methods Lovo cell were cultured in vitro,and this study was divided into control group,simple contrast agent group,microbubble plus ultrasound radiation group and simple ultrasound radiation group.Ultrasound contrast agent SonoVue and ultransound (frequency 1.5 MHz and MI 1.7) were used.Results In the control group,the microtubes were densest in dyeing silk,and extended to the edge of a cell.In the ultrasound and microbubble group,the microtubes were weaker and thin,the.network was mainly in the structure of the long axis.The cells arranged in a slight pigmentation to the density cell,to reach out many fine short hair,there were obviously a sense of silk and directional.In the microbubble and ultrasound group,Lovo cells streamed of the central significantly reduced,fluorescent lighting,with short hair.There was no difference in the ultrasound group and the microbubble group with microfilament and microtubule.Conclusions High mechanical index ultrasound radiation can changed into vacant,a trace of the assembly and distributed to the tumor cells are attacking,the transfer of restraining.

Objective To observe the perfusion pattern of hepatic angiomyolipomas using contrastenhanced ultrasound(CEUS) and compare diagnostic efficacy of contrast-enhanced ultrasound with contrastenhanced helical CT(CECT).Methods Nineteen patients with 21 resected and pathologically proven hepatic angiomyolipomas were included in this study.Low mechanical index (mechanical index less than 0.2) realtime CEUS was performed in nineteen patients (5 patients with pulse inversion harmonic, 14 patients with contrast pulse sequencing, CPS) after 2.4ml bolus injection of contrast agent SonoVue.CECT was performed using 16-slice helical CT and contrast agent Ultravist.The diagnostic performance was calculated by considering histologic results as the reference standards.Results Fifteen tumors were correctly diagnosed as hepatic angiomyolipomas,4 tumors were misdiagnosed (including 1 hepatocellular carcinoma,2hepatic adenomas, 1 hemangioma) and 2 tumors were characterized as benign lesions by CEUS.Eight tumors were correctly diagnosed as hepatic angiomyolipomas, 13 tumors were misdiagnosed (including 7hepatocellular carcinoma, 1 liposarcoma, 2 lipomas, 1 hepatic adenomas, 2 hemangiomas) by CECT.The preoperative diagnostic accuracy was 71.4% for CEUS and 38.1 % for CECT(P＜0.05 ).Conclusions CEUS can demonstrate typical imaging characteristics of most hepatic angiomyolipomas, and has higher diagnostic performance than CECT in characterization of hepatic angiomyotipomas.

Objective To evaluate mid-term(5-6 years)clinical and radiographic outcomes of cementless acetabular prosthesis in total hip arthroplasty(THA)and discuss the factors affecting postop- erative effect.Methods THA with cementless acetabular was used to treat 139 cases(148 hips)from January 1998 to May 1999.A retrospective study was done to analyze clinical effect and X-ray manifesta- tion of 109 cases(116 hips)that were followed up.Clinical evaluation was based on Harris hip score. The radiographical evaluation was performed according to orthotropia and lateral images of hip joint to ob- serve acetabular migration and bone changes around the hip and measure the polyethylene wear rate and direction of the acetabular polyethylene wear.Results The mean Harris score before operation was 44 points(10-70 points)but that after operation increased to 92.4 points(80-100 points)at latest follow up.Radiographical evaluation showed one hip with focal osteolysis,mean linear wear rate of 0.15 mm/ year and mean 10.4?for angle with vertical line.Abduction angel of the acetabula was mean 46?,with 10.5 mm for vertical distance and 8.7 mm for level distance.There was no revision of a fixed acetabulum component because of pelvic osteolysis secondary to polyethylene wear.No acetabular component was re- vised because of aseptic loosening or acetabular component migration at the latest follow up.Conclu- sions Cementless acetabular prosthesis in THA can successfully have good fixation.A long term follow up for cementless acetabular prosthesis is necessary because of potential factors like polyethylene wear and osteolysis.

Objective To evaluate tear ferning changes of conjunctivochalasis.Design Prospective case study series.Partici- pants 30 patients(60 eyes)of conjunctivochalasis and normal subjects were selected.Methods The subjects were observed with gen- eral ophthalmic examination and tear fern test(TFT).Tear ferning was classified into 4 types.TypeⅠand TypeⅡare normal.TypeⅢand TypeⅣare abnormal.Main Outcome Measures The type of tear feming.Results TFT showed that tear ferning was de- creased in conjunctivochalasis group(TypeⅢand TypeⅣoccupied 61.7%).The difference between conjunctivoehalasis and normal control group was significant(P