Fragments of ? and ? genes were cloned into expression plasmid pET-21b. Bacteria harbouring expression plasmid was induced with IPTG,and a band of expressed protein of 16 kD was found in PAGE. They expressed as soluble products. ? expression product reached about 5% of total bacterial protein,? expression product reached about 15% of total bacterial protein. The products were confirmed by Western-blotting.

Gastric cancer has high heterogeneity,and the traditional histopathologic classification has a limited significance for clinical work.Along with the dramatic development of molecular detection technology,it may be much more helpful for the treatment and prognosis to use the molecular classifications.Recently,there have been many researches on the molecular classification of gastric cancer,such as Tan type,Lei type,clonality type,The Cancer Genome Atlas (TCGA) type,Asian Cancer Research Group (ACRG) type,and so on.All of these molecular classification methods have respective advantages and disadvantages.

Objective To optimize an extracting technology for Tongluo granule by orthogonal design. Methods Water-extracting fraction:with the weight of water-extracting fraction and paeoniflorin content as the indexes,extracting times,water volume and extracting time were screened by L9(34) orthogonal test,Alcohol-extracting fraction:with the weight of alcohol-extracting fraction and hesperidin content as the indexes,alcohol concentration,extracting times,alcohol volume and extracting time were screened by L9(34) orthogonal test. Results The optimal extraction conditions were as follows:water-extracting fraction: extracting 3 times with 12-fold water,1.5 hours for each time;alcohol-extracting fraction:refluxing and extracting 2 times with 10-fold 60% alcohol,1.5 hours for each time. Conclusion The results can provide theoretical basis for production of Tongluo granule.

The technique of double labelling with fluorescent tracers(FB-NY,PI-Bb)has.been used in the following three groups of experiments:1.in 5 rats,one fluorescenttracer was injected into the tibial nerve and the other was injected into the peronealnerve;2.in 14 rats,one fluorescent tracer was injected into the sural cutaneousnerve and the other was injected into the branches to the soleus and gastrocnemiusmuscles;3.in 13 rats,one tracer was injected into the tibial nerve and the otherinto the bladder wall.The sections of the L_(3-6),S_(1-3) dorsal root ganglion were stu-died with a olympus fluorescence microscope using UG-1 excitation filter system.In the first and second groups,double labelled neurons were observed in L_(4-6) dorsalroot ganglion.But in the third group,the double labelled neurons were found onlyin the L_6 segment.The type of the double labelled neurons was mainly small-sized(33.1%)and small-medium sized(40.9%).The findings indicated that,the primary afferent fibers branched towards theperiphery and supplied,with different branches,either two somatic areas or skinand deep structures,or,especially,visceral and somatic sensory fields.This resultsuggests that the peripheral dichotomization of these dorsal root ganglion cells mightconverge sensory inputs from the somatic(tibial nerve)and the visceral(bladderwall)fields and thus provides one of the structural basis for the referred pain andthe neuronal mechanism of acupunctural therapy by which stimulation of somaticstructures could regulate the activities of visceral organs.

OBJECTIVE:To establish a method for the content determination of L-epicatechin in Actinidiae arguta. METHODS:HPLC was performed on the column of Kromasil C18 with mobile phase of acetonitrile-0.2%Acetic acid solution(15:85,V/V)at a flow rate of 1.0 ml/min,detection wavelength was 210 nm,column temperature was 25 ℃,and the volume injection was 10 μl. RESULTS:The linear range of L-epicatechin was 10.47-167.52 μg/ml(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 2%;average recovery was 98.07%-101.71%(RSD=1.39%,n=6). CONCLUSIONS:The method is simple, accurate and reliable,and suitable for the content determination of L-epicatechin in A. arguta.

Objective:To review the chemical and pharmacological activities of Codonopsis lanceolata in order to provide reference for the further development of C. lanceolata. Methods:The related literatures at home and abroad in the past 40 years were reviewed and analyzed, and then the chemical components and pharmacological actions of C. lanceolata were summarized. Results: The major chemical constiturents in C. lanceolata were terpenoids, alkaloids, phenylethanoid glycoside and flavonoids. The pharmacological ac-tivities were antioxidation, anti-inflammatory, anti-tumor, antiplatelet aggregation, hypoglycemic and hypolipidemic effects, etc. Con-clusion:The review provides reference for the further development and comprehensive utilization of C. lanceolata. The development of relevant safe and effective agents is still needed, and at present, the definition of mechanism and the extension of clinical application remain as the primary tasks of the exploration of C. lanceolata.

Objective: To optimize a preparation technology for pyretic arthralgia cataplasma. Methods: The preparation technology were studied by a uniform design experiment in which NP-700, tartaric acid, PVP, dihydroxyaluminum aminoacetate, glycerol, water and medicinal powder were factors and viscosity, infiltration, gel mobility and gel strength were indices. Results: The best ratio of this cataplasma matrix was NP-700:tartaric acid:PVP:dihydroxyaluminum aminoacetate: glycerol:water:powder = 4.0:0.2:1.0:0.1:25.0:35.0:2.0. According to optimized formula, to prepare the poultice, then to spread the poultice uniformly onto non-woven fabrics, cover CPP membrane and pack after 1 week at room temperature. Conclusion: Pyretic arthralgia cataplasma was well moldable and its process technology was feasible.

Objective To screen down regulated genes and find down regulated novel genes in gastric cancer. Methods Genes mRNA expression were detected between gastric cancer and normal gastric mucous membrane of five patients using cDNA microarray. Genes mRNA expression signals on hybridization membranes were analysized with computer software. Down regulated genes in gastric cancer were screened. cDNA suppression subtraction library was established by counterpart normal gastric mucous membrane mRNA(Tester) subtracting gastric cancer tissues mRNA(Driver) of five patients. After identification of the subtraction library, positive clones choosen randomly were sequenced , and down regulated novel genes in gastric cancer were screened. Some of the genes were identified by RT PCR.Results Down regulated genes in gastric cancer consist of 60 genes including tumor suppressing genes, apoptosis related genes, DNA replication and transcript or translate related genes, cell cycle related genes, cell migration related genes, etc. Two unknown gene fragments in gastric cancer were cloned. Conclusions Sixty down regulated genes in gastric cancer are confirmed. They are involved in gastric tumorigenicity and metastasis. cDNA subtraction library of gastric cancer was constructed successfully. Two down regulated novel gene fragments in gastric cancer was found.

OBJECTIVETo study the regularity of the passive tensile of the masticatory muscles and ligaments by Twin-Block appliance under various bite reconstruction, and to provide some biomechanical references for the clinical use and improvement of Twin-Block appliance.

METHODS"Temporomandibular joint, mandible and Twin-Block appliance" model was set up by the three dimensional finite element method, and the related masticatory muscles and ligaments were added on it. Seven experimental groups were designed according to the clinical and research, the occlusal inclined plate's angles of Twin-Block appliance were 40 degrees, 45 degrees, 50 degrees, 55 degrees, 60 degrees, 65 degrees and 70 degrees. The passive tensile in the masticatory muscles and ligaments were analyzed by the computer.

RESULTSUnder various experimental groups, the passive tensile in the anterior deep masseter (AM), the posterior deep masseter (PM), the anterior temporalis (AT), the posterior temporalis (PT), the stylomandibular ligament and sphenomandibular ligament improved with the increased slant angles of occlusal guide. The maximum value of the passive tensile was 82.57 N, the minimum value was 0.07 N.

CONCLUSION1) In various experimental groups, AT, AM, PM, PT, stylomandibular ligament and sphenomandibular ligament are subject to passive tension force in the process of Twin-Block appliance guiding the mandibular forward and play the important role on the remodeling of the mandible. 2)All groups of occlusal inclined plate's angle are in physiologically tolerable range and can be used in clinic.

Percutaneous nephrolithotomy is one of the common surgical modalities for the treatment of upper urinary calculi. In recent years, with the continuous development and improvement of endoscopic technology, laser technology and negative pressure suction technology, stone free rate of minimally invasive percutaneous nephrolithotomy is improved and the operation time is shortened, making the patient trauma smaller, comfort increased and hospitalization time shortened. This positive surgical effect and reliable safety have enabled minimally invasive percutaneous nephrolithotomy to develop rapidly and become widely used in the clinic. In this paper, the application and progress of minimally invasive percutaneous nephrolithotomy in the clinic are reviewed, with a view to providing a reference basis and clinical guidance for future clinical treatment.