Objective The objective of this study was to investigate the relationship between the expression of RAD18 and radiation resistance in glioblastoma multiforme(GBM)and to provide a new therapeutic target for improving the radiation resistance of GBM.Methods Human glioma A172 cells were transfected into blank and RAD18-containing plasmid vector.The cell proliferation of two groups after the same dose radiation was detected by cloning assay.The mRNA expression of RAD18 in primary and recurrent GBM samples after close proximity treatment were detected by qRT-PCR.All data were analyzed statistically.Results The proliferation of GBM cells transfected with RAD18 plasmid was higher than that of cells transfected with blank plasmid after radiation therapy(P<0.001).The expression level of RAD18 mRNA in recurrent GBM was higher than that in the untreated radioactive granules primary GBM(P<0.01).Conclusion The resistance of recurrent GBM to radiotherapy may be associated with the overexpression of RAD18 protein.

Objective To study the value of MSCT angiography(MSCTA)in diagnosing lower extremities arterial disease (LEAD) of diabetic foot ulcers and analyze the possible influential factors on the onset of diabetic foot ulcers.Methods 80 diabetic patients for MSCTA examination were divided into two groups by whether combined with foot ulcers.The tibial artery calcification score (TACS) and peripheral arterial occlusion index (PAOI) were calculated and the possible risk factors of diabetic foot ulcer were analyzed.Results There were 30 cases suffered with foot ulcer and 18 cases occurred lower extremity arterial occlusion (LEAO), the average PAOI was 4.40(3.60, 5.75).Non-foot ulcer cases were 50, and there were 8 cases suffered with LEAO, the average PAOI was 2.05(1.43, 3.10).Compared to patients without foot ulcer, patients with foot ulcer were older, had a lower BMI, and were more likely to have a history of tobacco use.They usually had higher TACS and severer peripheral arterial occlusive disease (PAOD).Stepwise regression model showed that foot ulcers had significant correlation with TACS and PAOI.Partial correlation analysis showed TACS was independent from PAOI in impacting diabetic foot ulcer.Conclusion MSCTA has important clinical value in diagnosing and valuing diabetic foot ulcer LEAD.Age, smoking history,BMI,TACS and PAOI affect the incidence of diabetic foot ulcers, and TACS and PAOI are the independent risk factors for diabetic foot ulcers.

In modern biology and biotechnology research, recombinant gene expression has been the most popular method to obtain the target protein. In recent years, many foreign genes have been efficiently expressed in Escherichia coli. However, proteins encoded by animal, plant or mesophilic microbial genes often lose activities or become denatured within a few hours at regular growth temperatures for E. coli; some other target proteins are toxic to host cells and therefore difficult to be over-expressed. The new T-vector, pEXC-T, was constructed by combining TA cloning and cold-shock induction to obtain high expression levels with low costs. This paper reports the construction of pEXC-T and optimization of induction techniques for gene expression. Two instable proteins were tested and successfully expressed in soluble form by using pEXC vector. The development of pEXC-T offers a convenient technique for the preparations of recombinant proteins to be used in structure/function studies, or as diagnostic markers and medicinal proteins.
Biotechnology ; Cold Temperature ; Escherichia coli ; genetics ; Gene Expression ; Genetic Vectors ; Plasmids ; genetics ; Recombinant Proteins

Objective To explore CTA characteristics of diabetic lower extremities arterial disease (LEAD) with different Fontaine stage.Methods Data of 100 patients clinically diagnosed as diabetic LEAD and underwent lower limbs arterial CTA were analyzed retrospectively.The patients were classified into grade Ⅰ-Ⅳ according to Fontaine classifications.Lower extremities arteries were graded according to CTA.Stenosis degree of lower extremity arteries in different Fontaine stages were analyzed.Results The grades of lower extremity arteries stenosis in different Fontaine stages had statistically significant difference (x2 =186.24,P＜0.001).There had statistically significant difference of stenosis degree in different lower extremity arterial levels among different Fontaine stages (superior genicular artery:x2 =69.24,P＜0.001;inferior genicular artery:x2 =111.59,P＜0.001;dorsalis pedis and arteriae plantaris:x2 =94.15,P＜0.001).Grades of stenosis between superior genicular arteries and inferior genicular arteries,superior genicular arteries and dorsalis pedis and arteriae plantaris had statistically significant differences (Z=12.59,P＜0.001;Z=10.47,P＜0.001).There was statistically significant difference of stenosis grades between inferior genicular artery and dorsalis pedis and arteriae plantaris (Z=12.66,P＜0.001).Occluded inferior genicular arteries usually associated with collateral vessels.Conclusion CTA can reflect the clinical stages of LEAD,which is the ideal method in diagnosing of diabetic LEAD.

In recent years,medical colleges have generally accepted the higher education assessment(evaluation).How to put the good practices and experience we obtained during the educational assessment into the daily teaching management and achieve normalization of teaching evaluation is of great significance to continuously improving the teaching quality.It is the primary condition to establish the evaluation index system for evaluating clinical teaching quality by regular teaching assessment. In this paper,the author constructed a clinical teaching quality evaluation index system through systematic research and exploration.

Tibial plateau fracture is a common acute trauma of the knee joint. At present, there are many studies on its classification and treatment, and minimally invasive treatment has become a research hotspot and mainstream direction of tibial plateau fracture. We summarized the clinical results of minimally invasive treatment of more than 300 cases of tibial plateau fractures, and proposed the concept of core weight-bearing area on tibial plateau, that is, the core weight-bearing area of the tibial plateau of the knee joint under normal motion statuswhile walking and moderate-intensity running. We performed thinsection CT scanning of the knee joint in a male volunteer for three-dimension finite element modeling.The results showed that during the walking state (the load was twice that of gravity), the core weight-bearing area of the medial and lateral plateaus was 389 mm 2 and 363 mm 2, accounting for 33.2% and 42.9% of tibial plateau, respectively;during the moderate-intensity running state (the load was four times that of gravity), the core weight-bearing area of the medial and lateral plateaus was 418 mm 2 and 406 mm 2, accounting for 35.6% and 48.0%of tibial plateau, respectively. Accordingly, tibial plateau fractures are supposed to be divided into core weight-bearing fracture and non-core weight-bearing fracture, and there are significant differences in the treatment ofthese twokinds of fractures: reduction is more demanding for core weight-bearing fracture,and the fracture involves the core area closely, the anatomical reduction is sought; for non-core weight-bearing area, the reduction requirements can be appropriately low demanded, and even in some cases , for example simple avulsion fracture, marginal fracture, some tibial plateau Hoffa fractures,can be treated conservatively. In summary, during clinical diagnosis and treatmentpractice, orthopedic surgeons should take the core weight-bearing area fracture as the core of diagnosis and treatment, strictly evaluate the extent of fracture involvement, select targeted internal fixation materials, and target to promote more accurate, minimally invasive, and individualized treatment of tibial plateau fractures.

Objective: To investigate the effect of RNA interference (RNAi) targeting AKT1 and PI3K P85 on the proliferation and invasion of breast carcinoma MCF-7 cells. Methods: The recombinant adenovirus expression vector, which contained short hairpin RNA (shRNA) targeting open reading frames of AKT1 and PI3K P85 (rAd5-siAKT1-siPI3K), was transfected into human breast carcinoma MCF-7 cells. AKT1 and PI3K P85 mRNA and protein expressions were detected by real-time PCR and Western blotting analysis. The expressions of PCNA, cyclinD1, and P53 were also detected by Western blotting analysis. The proliferation and apoptosis of MCF-7 cells were measured by MTT, flow cytometry and 2-dementinal and 3-dementional matrigel assay. Results: Recombinant adenovirus vector rAd5-siAKT1-siPI3K dramatically down-regulated AKT1 and PI3K P85 mRNA and protein expressions in MCF-7 cells; the downstream factors PCNA and cyclin D1 were also down-regulated, while P53 was up-regulated. Growth of MCF-7 cells was inhibited by over 50% in rAd5-siAKT1-siPI3K group as measured by MTT assay, and cell cycle was arrested in G_1/G_0 phase compared with untransfected and rAd5-siCtrl transfected groups. Cell growth on matrigel matrix showed normal cell shapes, while the cells in rAd5-siAKT1-siPI3K transfected group were detached from the matrix or grew in scattered clustering patterns, forming only small aggregates. Conclusion: shRNA targeting AKT1 and PI3K P85 can significantly down-regulate the expression of AKT1 and PI3K P85 in breast carcinoma MCF-7 cells, and inhibit the growth of MCF-7 cells in vitro.

Objective: To study the effect of and possible mechanism of knockinng down PI3Kp85α using siRNA in MCF-7 human breast cancer cell line. Methods: Oligofectamine was used to transfect PI3Kp85α siRNA to knock down the PI3Kp85α expression level in MCF-7 human breast cancer cell line in vitro. Real-time PCR was conducted to detect the expression of PI3Kp85α. The effect of PI3Kp85αsiRNA on the growth of MCF-7 cells was measured by MTT. The cell cycle distribution and cell apoptosis were detected by cell flow cytometry. Protein expression was evaluated by immunofluorescence staining and Western blot. Results: The expression of PI3Kp85 α was knocked down with PI3Kp85α siRNA in MCF-7 cells. Cell growth was delayed in PI3Kp85αsiRNA-treated group. Conclusion: The suppressive effect of PI3Kp85αsiRNA on the growth of MCF-7 human breast cancer cell line is significant and PI3Kp85α could be a candidate for gene therapy for breast cancer.

Objective To compare and analyze the epidemiological features of adult patients with distal femoral fracture between the east and west areas in China from 2010 to 2011.Methods The data of adult patients with distal femoral fracture treated from January 2010 through December 2011 in 63 hospitals were collected through the PACS system and case reports checking system.The data from 35 hospitals in the east area were classified as group A and those from 28 hospitals in the west area as group B.The analytic items included gender,age and AO classification.Results A total of 2,523 adult distal femoral fractures were collected,involving i,544 males and 979 females,with a male to female ratio of 1.58:1.The adult femoral fractures predominated in an age range from 41 to 50 years (18.94％) and their high-risk type was 33-A (50.18％).In group A of 1,650 cases,there were 1,027 males and 623 females,with a male to female ratio of 1.65:1 and a median age of 48 years;in group B of 873 cases,there were 517 males and 356 females,with a male to female ratio of 1.45:1 and a median age of 45 years.There were no significant differences in the age distribution and gender proportion between groups A and B (P ＞ 0.05).The age distribution showed that the peak ranges were from 31 to 60 years in both groups.The proportion of type 33-A was the most and that of type 33-B the least in both groups.The proportion of type 33-A in group B (53.49％) was significantly higher than in group A (48.42％) while that of type 33-B in group B (16.84％) was significantly lower than in group A (21.39％) (P ＜ O.05).Conclusions Adult distal femoral fractures were common in middle-aged males and their high-risk type was 33-A.Their peak age was from 31 to 60 years in both east and west areas in China.They were more common in men.The predominant fracture type was 33-A in both areas.The east area witnessed a significant higher proportion of type 33-B and a significant lower proportion of type 33-A than the west area.

OBJECTIVE: To evaluate the safety and effect of foley catheter traction for hemorrhage after postmicrochannel percutaneous nephrolithotomy (mPCNL). METHODS: Eighty-eight patients with upper urinary calculi were collected prospectively at the Department of Urology of Xiangya Hospital of Central South University from November 2010 to June 2011. The patients underwent mPCNL, and were divided into 2 groups randomly: 45 patients with 16F foley catheter but without traction served as the control group, and the other 43 patients with 16F foley catheter traction served as the experiment group. Blood loss was estimated by the mass of hemoglobin in the draining liquid and urine during postoperative duration through the HiCN. The blood loss and bleeding time were compared in the 2 groups, and analyzed by Wilcoxon rank sum test. RESULTS: There was statistical difference in the average blood loss between the control group (13.830 g) and the experiment group (7.959 g, P<0 .001). The mean bleeding time was 4 and 3 days in the control group and the experiment group respectively. CONCLUSION Foley catheter traction for mPCNL can reduce the blood loss, suggesting that Foley catheter traction is safe, effective and feasible.
Adolescent ; Adult ; Aged ; Blood Loss, Surgical ; prevention & control ; Child ; Female ; Hemostatic Techniques ; Humans ; Kidney Calculi ; surgery ; Male ; Middle Aged ; Nephrostomy, Percutaneous ; adverse effects ; methods ; Postoperative Hemorrhage ; therapy ; Prospective Studies ; Traction ; Ureteral Calculi ; surgery ; Urinary Catheterization ; Young Adult