Objective To construct and identify the incompetent-replication adenovirus carrying the fusiongene composed of the encoding gene of prepropeptide of mouse nerve growth factor(PN)and human beta-endorphin(?-EP)geue.Methods The gene segments of PN obtained from total RNA of the submandibular glandof a 2-week old Kumning mouse were amplified by RT-PCR and joined with the segment of ?-EP to form the fusiongene which was sequenced.The fusion gene contained in the incompetent-replication adenovirus was formed in theBJ-Ad Easy-1 susceptible cells and identified by PCR so as to choose the positive clone without wild vectors.Thecorrect clone was amplified and purified.The titers of adenovirus were determined using the specific 50% tissueculture infection dosage(TCID 50)method.Three days after the adenovirns was transferred into the cultured A431cells,RT-PCR was performed to showed the transcribed mRNA of this fusion gene and the intracellular ?-EPexpression was quanlitatively detected by inummo-histological method.Finally the concentration of human ?-EP inthe culture medium was determined by quantitative radio-immunoassay on 1st,3rd and 7th day afterinfeetion.Results The sequence of the fusion gene was correct.The titer of recombinant adenovirus Ad-NEP was1.5?10~10 pfu/ml.Three days after infection a 475 bp segment was amplified by RT-PCR and abundant orangegranules were shown in the infected cell.The ?-EP concentration in the culture medium was significantly higher inAd-NEP group than in the control group on 1st,3rd and 7th day(P

OBJECTIVETo investigate the diagnostic value of MRI in radial head fracture with forearm interosseous membrane injuries.

METHODSFrom December 2011 to December 2012,26 patients with fractures of capitulum radial in our hospital were collected. There were 15 males and 11 females, ranging in age from 21 to 53 years old,with an average of 37.6 years old. All the patients visited hospital within 72 hours after injuries. X-ray radiography of full ulnar radial length in injured side, CT in injured side (three-dimensional reconstruction if necessary) and MRI (including the elbow and wrist joints) were performed within a week after the injury. The MRI manifestations of the forearm interosseous membrane (with or without damage, the injured location and the injury degree ) and the fractures degree of radial head were observed and compared for the relativity.

RESULTSRadial head fracture from Mason type I to III was associated with the forearm interosseous membrane injury. Radial head fracture degree was positive correlated with forearm interosseous membrane injury degree (P < 0.05).

CONCLUSIONRadial head fracture with suspicious forearm interosseous membrane injury is necessary to take MRI for checking for any interosseous membrane injury and injury degree, then choose the right treatment for radial capitulum fracture, only in this way can be helpful for the functional recovery of elbow and forearm.

Adult ; Female ; Forearm ; pathology ; Humans ; Magnetic Resonance Imaging ; Male ; Membranes ; injuries ; Middle Aged ; Radius Fractures ; diagnosis ; pathology ; Young Adult

OBJECTIVETo observe the clinical effect of Guilu Erxian Glue Cataplasm (GEGC) on carcinoma of the large intestine patients with myelosuppression after chemotherapy, and further to confirm its efficiency and safety.

METHODSTotally 60 patients with carcinoma of the large intestine were randomly assigned to two groups. Meanwhile, they all accepted FOLFIRI chemotherapy. Patients in the treatment group were additionally applied at Shenque (RN8), exchanging once per every other day, for 14 successive days. Patients in the control group took placebos with the same dose and dosage as the treatment group. The blood cell counts (WBC, NE, and PLT) were detected before chemotherapy, at day 7, 10, and 14. The TCM symptoms integrals, Karnofsky performance score (KPS), liver and kidney functions were observed before chemotherapy, at day 7 and day 14. Adverse skin reactions were observed each day. And the usage of hematopoietic growth factors was recorded.

RESULTS(1) The KPS score at day 7 was more stable in the treatment group than in the control group; the WBC and NE counts in the peripheral blood at day 14 were higher in the treatment group than in the control group; and TCM symptoms integrals at day 14 was lower in the treatment group than in the control group, all with statistical difference (P < 0.05). (2) Compared with the control group, the PLT count was higher in the treatment group than in the control group, the usage of rhG-CSF and antibiotics was less in the treatment group than in the control group, all with no statistical difference (P > 0.05). (3) No obvious adverse reactions such as liver injury, renal injury, or skin allergy were observed.

CONCLUSIONSAdjuvant treatment of GEGC could improve carcinoma of the large intestine patients with myelosuppression to some extent. No relevant adverse reactions were found.

Adjuvants, Immunologic ; therapeutic use ; Adult ; Aged ; Bone Marrow Diseases ; chemically induced ; drug therapy ; Colorectal Neoplasms ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged

Objective To investigate the effect of mesenchymal stem cells (MSC) on the activity of nuclear factor (NF)-?B in rats with myocardial infarction.Methods MSC were isolated from SD rats (120—150 g in weight).SD rats (180—200 g in weight) were subjected to MI by left coronary artery occlusion,and were allo- cated into three groups randomly:1)sham group (without ligation of the artery,n=8);2)injection of PBS solu- tion (n=8);3)injection of MSC (n=8).MSC or PBS solution was injected into myocardium from epicardium instantly after MI models were established.Four weeks after transplantation,cardiac function was evaluated u- sing physiological recorder.Western blot were performed to investigate the nuclear factor-? activity.The ex- pressions of tumor necrosis factor (TNF)-? and interleukin (IL)-6 were evaluated by RT-PCR and Western blot. Results 1)Mortality was 20%(2/10) in sham group,33.3%(4/12) in PBS group and 20%(2/10) in MSC group with no statistic differences between them(P=0.646).2) Hemodynamic measurements showed that MSC trans- plantation caused significant improvement in cardiac function,comparing with MI+PBS group.3) MSC inhibi- ted the activities of NF-?B in myocardium and down-regulated the expression of TNF-? and IL-6 in mRNA and protein level.Conclusion Transplantation of MSC improved cardiac function in MI rats,which may partly at- tribute to their immuno-inflammatory regulation mechanism.

Epidemiologic Methods ; Epidemiology ; Humans ; Terminology as Topic

Objective To investigate the efficacy and accuracy of 3D printing individualized navigation templates used in shoulder hemi-arthroplasty (HA).Methods We collected 9 adult cadaveric specimens of 18 shoulders with no obvious deformities or defects.After CT scanning was conducted on bilateral shoulders,the raw CT data in DICOM format were imported into software Mimics 10.01 and Imageware 12.0 to reconstruct 3D shoulder models.The retroversion angles and heights of bilateral humeral heads were measured to design individualized navigational templates using software Geomagic Studio 12.0.Rapid prototyping (RP) technique was used to manufacture the individualized navigation templates which were then used to assist shoulder HA in cadaveric specimens.CT scanning was performed postoperatively to measure the retroversion angle and height of the prosthesis implanted for comparison with the preoperative measurements.Results Before shoulder replacement in the 9 cadaveric specimens,respectively,the heights of left and right humeral heads averaged 300.0 ± 11.6 mm and 300.0 ± 11.4 mm,and the retroversion angles 15.6°±4.9° and 15.9°± 4.9°.After the replacement,the heights of left and right humeral heads averaged 300.0 ± 11.3 mm and 300.0 ± 11.7 mm,respectively,and the retroversion angles 15.7° ± 4.9° and 15.8° ± 5.2°,respectively.There were no significant differences between the left and right sides in the pre-or post-operative heights or retroversion angles of the humeral head (P ＞ 0.05).There were no significant differences either between preand post-operation in the heights or retroversion angles of the left and right humeral heads (P ＞ 0.05).The mean errors for the preoperative and postoperative heights and retroversion angles of the humeral heads were 1.44 ± 0.96 mm and 0.25° ± 0.15°,respectively.Conclusion Individualized navigation templates 3D printed can be used to assist effective and accurate shoulder HA because they are designed and manufactured according to accurate measurements of retroversion angles and heights of the humeral heads which are acquired by computer preoperatively.

P300/CBP-associated factor(PCAF),an important member of histone acetyltransferase family(HATs) within eukaryotic cells,is capable of inducing the acetylation of histone,promoting the transcription of specific genes and involving in many biological effects.In the present study,full-length cDNA of PCAF was inserted into plasmid pGEX-5x-1,then the soluble protein GST-PCAF was expressed in E.coli BL21(DE3) after the optimization of inducing conditions.The recombinant protein was further purified with affinity chromatography and tested the activity by in vitro acetylation assays.High efficient PCAF protein produced by this method could serve for the study on the role of PCAF in gene regulation and the interaction between PCAF and other proteins.

BackgroundIron-containing foreign body trapped in the eyeball wall without affecting the opticus occurs occasionally in clinic. Operation always is performed in an attempt to avoid the deposition of rust in different tissues of the eye-balls. However,a few animal experimental studies showed that a small foreign body does not affect the retina and opticus in the period of three months. The question of whether surgery needs to be carried out is worth discussion. ObjectiveThe aim of this study was to evaluate the influence of posterior intrascleral iron foreign body on the rabbit retina and opticus. MethodsTwelve healthy adult Japan flap-eared white rabbits were randomly divided into two groups. Medium carbon iron with rust or without rust( size of 2. 0 mm × 1. 0 mm×0. 2 mm) were implanted into the posterior sclera of the left eye to create the animal model with iron foreign body in the eyeball wall. The cornea, anterior chamber, crystalline lens, vitreous and fundus of the rabbits were observed under a slit lamp microscope 1weekbeforeoperationand 1week, 2weeks, 1monthand 3months after operation.Flash electroretinogram(F-ERG) and flash visual evoked potential (F-VEP) were recorded at the time points mentioned above. All the rabbits were sacrificed and the eye balls were extracted at the end of the experiment, and the position of the iron foreign body was determined. The histopathological examinations of the retina and opticus were performed under the light microscope. This experiment complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. ResultsThere were no statistical differences for the a-wave amplitude of F-ERG among different time points( F =1. 885,P =0. 129 ) and different treatment groups ( F =1. 188, P =0. 340 ), as with the ERG b-wave amplitude ( time: F =2. 73, P =0. 064 ; group : F =1. 114, P =0. 367). The differences in the latencies of F-VEP N1-wave were insignificant among the different time points( F =1. 605, P =0. 263 ) as well as various groups ( F=1. 556, P =0.314 ), and those of F-VEP P1 -wave were not evidently changed ( time: F =2. 329, P =0. 092 ; group : F =2. 186, P =0. 103 ). No correlations were seen between the time factor and grouping factor ( P ＞ 0. 05 ). There was no apparent siderosis bulbi change during the follow-up duration. No morphological abnormality in the retina and optical nerve was found under the light microscope. At the end of the experiment,intrascleral iron foreign body was wrapped by surrounding tissue in a stable condition. Conclusions The small posterior intrascleral iron foreign body, whether it is oxidized or not, does not produce distinctive functional or pathological damage on retina and opticus in the short term.

Objective：This study was designed to investigate BCRP gene expression in normal lung and in non-small cell lung cancer tissue. Methods：RNA was extracted immediately from the fresh normal lung tissue and viable tumor tissue harvested from surgically resected specimens of non-small cell lung cancer patients. cDNA of BCRP gene was prepared by RT-PCR and then amplified by PCR. The cDNA products from those specimens were transferred to blotting membrane through electrophoresis and transferring technique. Southern blot hybridization was eventually performed to detect the expression of BCRP gene. Results：RNAs were extracted from 8 tumor tissues and 12 pairs of tumor tissue and normal lung tissue were harvested from the same lung. Four patients’RNA samples with poor quality due to degrading were discarded. cDNA products of BCRP gene were obtained by RT-PCR and amplified by PCR in the remain 16 patients’RNA samples. Through Southern blot hybridization, BCRP gene was found to have slight expression in various amounts in all normal lung tissue(10/10) and only in half of tumor tissue samples (8/16). Conclusion：BCRP gene is slightly expressed in a variable amount in all normal lung tissue and only in half of tumor tissue for this group of patients with non-small cell lung cancer. Therefore chemotherapy with anthracycline may possibly induce the overexpression of BCRP gene and at last to develop multidrug resistance.

OBJECTIVETo establish a method for assist design of the posterior pharyngeal flap (PPF).

METHODSThe width, length of velopharyngeal space in repose and phonation of "i" in 26 cases with VPI were measured with CT. The width and length of PPF needed, the location of the pedicle and the contact position with the soft palate were analyzed.

RESULTSThe width of velopharyngeal space in repose ranged from 15.10 to 37.25 mm, with the average of 27.01 mm, the length was 12.54 to 30.39 mm, with the average of 22.10 mm. The width in phonation of "i" was 9.11 to 34.06 mm, with the average of 17.69 mm, the position of the pedicle wasall located at the level of the first cervical vertebra or above and the contact area of the flap with the soft palate was in the middle posterior 1/3 mostly.

CONCLUSIONAs a method to quantitatively evaluate velopharyngeal function, CT can assist in design for the PPF.

Adolescent ; Adult ; Child ; Cleft Palate ; diagnostic imaging ; surgery ; Female ; Humans ; Male ; Pharynx ; diagnostic imaging ; surgery ; Surgical Flaps ; Tomography, X-Ray Computed ; Velopharyngeal Insufficiency ; diagnostic imaging ; surgery