TSH1, a novel transcription factor in rice containing conserved AP2/EREBP domain, was screened from the EST database. The specific interaction of TSH1 and DNA elements was detected by two kinds of traditional methods, electronic mobility assay and yeast one-hybrid assay, which suggested that THS1 specifically binds to GCC element in vitro and in vivo. Furthermore, It was confirmed by atom force microscopy (AFM) that TSH1 binds to GCC element (core sequence GCCGCC) not DRE element (core sequence GCCGAC), and the single molecular force between TSH1 and GCC element was quantitatively measured. The molecular force of TSH1 with GCC element is (83.9 ?2.2) pN. This interaction can be observably blocked by the dissociate TSH1 protein in the solution. These above results proved that TSH1 recognizes and binds to GCC with specifically interaction, which also demonstrate the high sensitivity of the AFM measurements for the single base substitution of the GCC core sequence greatly reduced the binding affinity. Comparing these methods for identification the binding of transcription factor with its DNA element, it was considered that AFM is expected to be a sensitive and reliable method that offers valuable information for the characterization of transcription factors.

BACKGROUND:The anatomic structures and kinetic characteristics are the bases to establish hand model, and the kinetic characteristics of hand are determined by the anatomic structure. So, numerous scholars have paid close attention to virtual hand models based on the anatomic structures of hand.OBJECTIVE: To construct visible hand model based on anatomic structure.DESIGN: Single sample trial.SETTING: Center Laboratory, Third Military Medical University of Chinese PLA.MATERIALS: This trial was carried out in the Center Laboratory, Third Military Medical University of Chinese PLA in October 2003. Two fresh adult hands, which involved wrist joint, provided by the Department of Anatomy, Third Military Medical University of Chinese PLA, were employed. The two hands had no organic damage by naked observation.METHODS: The specimens were embedded and mill-cut (mill-cut layer thinness 0.2 mm). Cannon (ESO 1OD) digital camera (6.3 million pixel) was used for image collection. Each image was 31.5 MB, Data of 1 200 images were obtained.Adobe Photoshop 7.0 software was used for image treatment and then two-dimensional cross-section images were collected. The bone, flexor tendon and the outline of hand was three-dimensionally reconstructed by using the software,which was developed by the Department of Computer Science and Technology,Tsinghua University and Institute of Computer Medicine, Department of Anatomy, Third Military Medical University of Chinese PLA.MATN OUTCOME MEASURES: Three-dimensional reconstruction of the bone, flexor tendon and outline of hand.RESULTS: ①The outline of hand: After being reconstructed, the outline, which consisted of all fingers and nails, was well displayed, and observed from many directions.② Three-dimensional reconstruction of the bone of hand: The reconstructed bones of hands involved digital bones, metacarpal bones, carpal bones and all joints, and they could be displayed solely or in groups with other reconstructed structures.③ Three-dimensional reconstruction of the flexor tendons of hand:Four superficial flexor tendons, four deep flexor tendons and one flexor pollex Iongus muscle tendon could be seen from the reconstructed images. All the flexor tendons went out together from the carpal canal and went through the palm, then spread out in fan-shape along the direction of each finger. ④ Three-dimensional reconstruction of the metacarpal fascial spaces: It could be observed from the reconstructed three-dimensional models that the proximal end of the thenar space was close, its distal end was open to the 1st web space, and the proximal end of the midpalmar fascial space was open to the posterior space of antebrachial flexor by carpal canal. Its distal end had three little spaces, which were open to the 2nd, 3rd and 4th web space, respectively.CONCLUSION: The visible hand model, which is preliminarily established, can precisely show the main anatomic structure of palm.

Cerebral microdialysis is a new sampling method that collects the perfusates from the specific brain region continuously. Disorders of extracellular excitatorary amino acids, monoamines and acetylcholine transmitters release cause brain injury during the cerebral ischemia and reperfusion. Microdialysis has been used to study the dynamic change of neurotransmitters during the acute cerebral ischemia and reperfusion lately, and the real change of neurotransmitters has been monitored in animals brain. The present paper reviews the progress on the microdialysis and its application in the study of neurotransmitters in cerebral ischemia animals in the world.

BACKGROUND:Some studies have shown that the double-bundle posterior cruciate ligament reconstruction is not superior to the single-bundle posterior cruciate ligament reconstruction, and stil has some deficiencies difficult to overcome. Which is better, double-bundle reconstruction or single-bundle reconstruction? There is no uniform conclusion. OBJECTIVE:To perform a prospective cohort study on the clinical efficacy and safety of autologous single- and double-bundle posterior cruciate ligament reconstruction. METHODS:Totaly 81 patients with posterior cruciate ligament injury were randomly divided into single-bundle reconstruction group (n=41) and double-bundle reconstruction group (n=40). The knee stability, Lysholm score, Tegner score, hospital stay, operation time, fever days and number of puncture cases were compared between the two groups before and 24 months after reconstruction. RESULTS AND CONCLUSION:Compared with the single-bundle reconstruction group, the knee stability was significantly worse in the double-bundle reconstruction group (F=4.362,P=0.000); the operation time, hospital stay and number of puncture cases were also higher in the double-bundle reconstruction group (P < 0.05). At 24 months after reconstruction, the Lysholm and Tegner scores were both increased significantly in the two groups (P < 0.05), but there was no difference between the two groups (P > 0.05). These findings indicate that both single- and double-bundle reconstruction under arthroscopy is safe and effective treatment for posterior cruciate ligament injury, but the double-bundle reconstruction is not recommended as the preferred surgical procedure because of longer time and larger trauma.

Objective To investigate the role of ?-catenin gene in breast tumorigenesis.Methods Mutation and expression of ?-catenin gene in 42 breast cancer tissues were detected by polymerase chain reaction-single strand conformation polymorphism and immunohistochemistry.Results The rate of abnormal expression of ?-catenin in breast cancer tissuse was 59.5%(25/42).Mutation of ?-catenin gene was not detected in breast cancer tissue.Conclusions Abnormal expression of ?-catenin plays an important role in human breast tumorigenesis,but the cause of abnormal expression of ?-catenin is not mutation of ?-catenin gene.

BACKGROUND: The metacarpal fascial spaces of hand are the potential structure that is not virtual, so that difficulties are presented in diagnosis and treatment of diseases of the metacarpal fascial spaces of hand. In order to solve the difficulties,anatomical borderline and abut relationship of the metacarpal fascial spaces of hand have to be sufficiently understood. OBJECTIVE: The borderline and abut of the metacarpal fascial spaces of hand were detailedly observed and researched by fresh cadaveric dissection, thin sectional anatomy and computer image recognition technology, and the 3-D reconstruction technology was utilized to reconstructed the metacarpal fascial spaces in this paper, so the abut relationship of the metacarpal fascial spaces of hand could be displayed, and the detailed anatomical data of imaging diagnosis and surgical treatments of disease of the fascial spaces of hand could be provided. DESIGN: Repeated measuring design.SETTING: The 251 Hospital of Chinese PLA and Central Laboratory of the Third Military Medical University of Chinese PLA. MATERIALS: Twelve adult fresh cadaveric hands which involved six right hands and six left hands and wrist joint, had no organic damage by naked observation, provided by the Department of Anatomy, the Third Military Medical University of Chinese PLA, among one right hand and one left hand were waiting for being mill-cut and thin cross-section dissected and imaging reconstructed. Among ten hands were observed by fresh cadaveric anatomy. METHODS: The trial was carried out in the 251 Hospital of Chinese PLA and Central Laboratory and Department of Anatomy of the Third Military Medical University of Chinese PLA from May 2003 to February 2006. Based on the gross anatomy, thin cross-section anatomy, visible human and virtual human technology theory, the metacarpal fascial spaces of hand were three-D reconstructed in microcomputer.MAIN OUTCOME MEASURES: Anatomical borderline and about relation of the metacarpal fascial spaces of hand. RESULTS: Display of observation result of adjacent and dimension of the midpalmar space and the thenar space by fresh cadaveric anatomy combined with thin cross-section anatomy, computer image recognition technology, and the 3-D reconstruction technology: The anterior borderline of the MPS was the digital flexor tendons of middle finger, ring finger, little finger and the 2nd lumbrical muscle (LM), 3rd LM, 4th LM; the posterior borderline was the palmar interosseous fascia before 3rd palmar bone, 4th palmar bone, 5th palmar bone and corresponding interosseous muscles; the lateral borderline was the palmar intermediate septum; the medial borderline of the MPS was the medial intermuscular septum of palm. In addition, the distal section of the MPS which was separated into three little spaces by two septums of connective tissue, and 3rd 4th, 5th digital flexor tendon and 2nd, 3rd, 4th LM were respectively contained by the three little spaces. The proximal of this space opened to the posterior space of antebrachial flexor by the carpal canal; the distal of this space opened to 2nd, 3rd, 4th web space (WS) by 2nd, 3rd, compartment of 4th LM, and continuously the dorsal subcutaneous space and the subaponeurotic space. The anterior borderline of the TS was the 1st digital flexor tendon and the 1st LM; the posterior borderline was the fascia of abductor pollicis; the medial borderline was the palmar intermediate septum (PIMS); the lateral borderline was the tenden sheath of flexor pollicis longus and the lateral intermuscular septum of palm. The distal of this space opened to 1st WS by compartment of the 1st LM, and continuously to the dorsal subcutaneous space and the subaponeurotic space; the proximal of the TS was close.CONCLUSION: The proximal of the thenar space is close, the distal section of the metacarpal fascial spaces is separated into three little spaces, and the computerized three-D renconstruction of the fascial spaces of hand can provide some guidance for imaging diagnosis and surgical treatments.

Objective To evaluate the role of autophagy in the development of inflammatory pain in the rats.Methods Twenty-four healthy adult male Sprague-Dawley rats,weighing 180-240 g,were randomly divided into 4 groups (n=6 each) by using a random number table:control group (group C),inflammatory pain group (group IP),dimethyl sulfoxide (DMSO) group (group D),and rapamycin (autophagy inducer) group (group R).Inflammatory pain was produced by injecting 50 μl bee venom into the plantar surface of the left hindpaw.In group C,0.9％ normal saline was injected into the plantar surface of the left hindpaw.In group D,2％ DMSO was injected through a gastric tube into the stomach 1 ml per day for 3 consecutive days,and the model was established at 1 h after injection on 3rd day.In group R,rapamycin l0 mg/kg (in 2％ DMSO) was injected through a gastric tube into stomach 1 ml per day for 3 consecutive days,and the model was established at 1 h after injection on 3rd day.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 2 h after the model was established.After measurement of the pain threshold,the dorsal horn of the spinal cord was removed for determination of the expression of microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ),Beclin-1 and p62 by Western blot.Results Compared with group C,the MWT was significantly decreased,the TWL was significantly shortened in IP and D groups,and the expression of LC3 Ⅱ,Beclin-1 and p62 in the dorsal horn of the spinal cord was significantly up-regulated in IP,D and R groups (P＜0.05 or 0.01).Compared with group IP,the MWT was significantly increased,the TWL was significantly prolonged,the expression of LC3 Ⅱ and Beclin-1 in the dorsal horn of the spinal cord was significantly up-regulated,and the expression of p62 was significantly down-regulated in group R (P＜0.05),and no significant change was found in the parameters mentioned above in group D (P＞0.05).Conclusion Autophagy disorders are involved in the development of inflammatory pain in the rats.

5-Flucytosine (5-FC) could be changed to 5-fluorouracil (5-FU) by cytosine deaminase (CD), the latter is able to kill cancer cells. However, there is no efficient method to deliver the CD gene into the tumor cells, which hampers the application of the suicide gene system. In this experiment, for the first time, the NDV has been utilized as a vector to deliver the CD gene into the cancer cells, the virus can infect the cancer cells specifically, replicate and assemble, while the cytosine deaminase is expressed. Then the CD converts the prodrug 5-FC into 5-FU to achieve the purpose of inhibiting tumor. Firstly, the whole genome of E. coli JM109 was extracted, and the CD gene was obtained by cloning method. Then the CD and IRES-EGFP were ligated into the pEE12.4 expression vector to become a recombinant pEE12.4IE-CD eukaryotic expression plasmid. The human liver cancer cells were transfected with the plasmid. The cells were treated with different concentrations of 5-FC, MTT method was used to determine the killing effect of CD/5-FC system on the human liver cancer cells. The cell deaths were 18.07%, 42.98% and 62.20% respectively when the concentrations of prodrug were at 10, 20 and 30 mmol x L(-1). In 5-FC acute toxicity experiment, Kunming mice were injected with different concentrations of 5-FC at intervals of 1:0.5. The LD50 of 5-FC through iv injection was determined by improved Karber's method, the LD50 was 507 mg x kg(-1) and the 95% confidence limit was 374-695 mg x kg(-1). According to the maximum LD0 dose of the LD50, the maximum safe dose was 200 mg x kg(-1). Body weight and clinic symptoms of the experimental animals were observed. These results laid the foundation to verify the antitumor effect and safety of CD/5-FC system in animal models. The CD gene was ligated into the NDV (rClone30) carrier, then the tumor-bearing animal was established to perform the tumor inhibiting experiment. The result showed that the recombinant rClone30-CD/5-FC system has a high antitumor activity in vivo. To summarize, CD gene has been cloned and its bioactivity has been confirmed in the mammalian cells. It is the first time in this study to utilize the recombinant NDV to deliver the CD gene into the tumor cells; our result proves the rClone30-CD/5-FC system is a potential method for cancer therapy.

Objective To develop a kind of rapid decompression equipment replacing the toughened glass simulating the state of aircraft cabin glass bursting on the fly.Methods The metallic membrane was used to isolate both chambers with different air pressures.The areas of decompression membrane and path were determined by calculating on the basis of aircraft decompression altitude,cabin pressure differential and decompression time.The structural strength was determined according to enduring force of the metallic membrane.The membrane was ejected by high pressure air using the ejection launch technology of aircraft missile.The result of simulating aircraft cabin glass bursting on the fly was achieved.Results The rapid decompression equipment ejected by air pressure in low-pressure chamber could achieve the state of simulating aircraft cabin glass bursting on the fly,and the best decompression time was 0.16 s.Conclusion The rapid decompression equipment ejected by air pressure accomplishes the decompression preparative in short time with easy operation,and can satisfy the desired requirements for the performance and precision.