Overexpresssion of NDUFA4 induces epithelial-mesenchymal transition of human colorectal cancer cells / 医学研究生学报

Objective The role and action mechanisms of NADH dehydrogenase 1 alpha subcomplex 4 (NDUFA4) in the development of human colorectal cancer (CRC) are not yet clarified. This article aims to study the effect of overexpressed NDUFA4 on the epithelial-mesenchymal transition (EMT) of human CRC cells. Methods The recombinant plasmid NDUFA4 (p-NDUFA4) and control plasmid (p-Cont) were transiently transfected into human CRC HCT116 cells. The expression of NDUFA4 in the cells was determined by real-time PCR and Western blot respectively, and the migration ability of the cells detected by Transwell migration and wound healing assays. The expression levels of MMP2, MMP9 and CXCR4 in the cells were measured by qRT-PCR, and those of Twist, Snail, E-cadherin and Vimentin by Western blot and by immunofluorescence assay, respectively. Results Compared with the p-Cont group, the human CRC HCT116 cells of the p-NDUFA4 group showed significantly up-regulated expressions of NDUFA4 mRNA ([0.96±0.15]% vs [1.94±0.08]%, P<0.05) and NDUFA4 protein ([0.06±0.05]% vs [1.07±0.12]%, P<0.05), increased in vitro migration ability ([29.51±3.17]% vs [54.36±4.08]%, P<0.01) and migrated cell rate ([0.99±0.12]% vs [1.85±0.10]%, P<0.01), elevated expression levels of MMP2, MMP9, CXCR4, N-cadherin,Vimentin, Snail and Twist, but down-regulated level of E-cadherin (P<0.05). Conclusion Overexpressed NDUFA4 promotes the epithelial-mesenchymal transition of human colorectal cancer cells.

Finite element analysis on the effect of lateral wedge insole intervention on the contact characteristics of the subtalar joint / 中国骨伤

<p><b>OBJECTIVE</b>To establish a three-dimensional finite element model of the lower limb bones, and investigate the changes of the contact characteristics of the subtalar joint after using laterally wedge insole intervention.</p><p><b>METHODS</b>Using the reverse modeling technology, the lower limb bones of normal adult volunteers was scanned by CT. Mimics 10.0 and Geomagic Studio 6.0 software were used to reconstruct the 3D morphology of bones and external soft tissue of the feet. The laterally wedge insole was designed in ProE 5.0. And then all the models were imported into Hyperwork 10.0 and meshed, and given the material properties. The finite element analysis was carried out in ABAQUS 6.9.</p><p><b>RESULTS</b>A three-dimensional finite element model of the lower extremity was established, which was consisted of 95 365 nodes and 246 238 elements. The contact area of the standing state of the lower joint was larger than that of the anterior middle joint surface. The peak stress was concentrated in the anterior lateral part of the posterior articular surface, and the average stress value was(3.85±1.03) MPa. Compared with the model of 0°, the contact area of the subtalar joint was reduced accordingly. There was a significant correlation between anterior middle joint ||=0.964,=0.008, and posterior articular ||=0.978,=0.002. The equivalent stress of 0° model distributed from(3.07±1.14) MPa to(3.85± 1.03) MPa, which had no statistically difference. Compared with the 0° model, the equivalent stress of the anterior and middle joint surfaces of the 8° model was significantly reduced(<0.05), but the peak stress of the posterior articular surface was significantly increased(<0.05). In the 12° model, the peak stress was sharply increased to(10.51±3.53) MPa. Compared with 8° model, there was no statistically difference(<0.05). Although the peak stress was slightly increased in 16° model, but compared with 12° model, there was no statistically differences(>0.05).</p><p><b>CONCLUSIONS</b>Although a certain valgus can be obtained in subtalar by wearing LWI, the result comes at the cost of the stress concentration on posterior surface. Through this study, we can find that LWI with 8° tilt angle could provide appropriate valgus moment without causing excessive concentration. Therefore, in order to avoid secondary ankle complications, we should not increase the tilt angle blindly.</p>

Effect of altitude chronic hypoxia on liver enzymes and its correlation with ACE/ACE2 in yak and migrated cattle / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the difference of liver enzyme levels and its correlation with serum ACE/ACE2 among yak and cattle on Qinghai-Tibetan plateau, and to further explore the biochemical mechanism of their liver of altitude adaptation.</p><p><b>METHODS</b>The serum samples of yak were collected at 3,000 m, 3,500 m, 4,000 m and 4,300 m respectively, meanwhile the serum samples of migrated cattle on plateau (2,500 m) and lowland cattle (1,300 m) were also collected. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), cholinesterase (CHE), gamma glutamyl transferase (GGT), alkaline phosphatase (ALP), serum lipase (LPS), angiotensin converting enzyme(ACE), angiotensin converting enzyme-2 (ACE2) in serum were measured by using fully automatic blood biochemcal analyzer. We analysed the differences of the above enzymes and its correlation with ACE/ACE2. We used one way analysis of variance (ANOVA).</p><p><b>RESULTS</b>The levels of ALT in 4,000 m group and 4,300 m group of yak increased significantly compared with other groups, there were no statistically significant differences in AST, CHE, GGT, ACE/ACE2 levels of yaks at different altitudes. As compared to lowland cattle, the serum levels of AST and CHE were increased, the level of LPS and ACE was decreased significantly, respectively, and especially, the ratio of ACE/ACE2 of migranted cattle reduced nearly two times. The levels of LPS were significantly correlated to the ratio of ACE/ACE2 in yak (r = 0.357, P < 0.01), and a high correlation between ALP and ACE/ACE2 in lowland cattle( r = 0.418, P < 0.05), But the biggest contribution rate of the ratio of ACE/ACE2 was only 17.5% for the changes of the levels of liver enzyme.</p><p><b>CONCLUSION</b>The results indicated that with the altitude increased did not significantly influence the changes of liver enzymes' activities in mountainous yaks but not in cattle. However, all above these changes weren't actually correlated to the ratio of ACE/ACE2.</p>

Alterations in cardiac structure and function in a modified rat model of myocardial hypertrophy / 华中科技大学学报（医学）（英德文版）

This study was aimed to establish a stable animal model of left ventricular hypertrophy (LVH) to provide theoretical and experimental basis for understanding the development of LVH. The abdominal aorta of male Wistar rats (80-100 g) was constricted to a diameter of 0.55 mm between the branches of the celiac and anterior mesenteric arteries. Echocardiography using a linear phased array probe was performed as well as pathological examination and plasma B-type natriuretic peptide (BNP) measurement at 3, 4 and 6 weeks after abdominal aortic constriction (AAC). The results showed that the acute mortality rate (within 24 h) of this modified rat model was 8%. Animals who underwent AAC demonstrated significantly increased interventricular septal (IVS), LV posterior wall (LVPWd), LV mass index (LVMI), cross-sectional area (CSA) of myocytes, and perivascular fibrosis; the ejection fraction (EF), fractional shortening (FS), and cardiac output (CO) were consistently lower at each time point after AAC. Notably, differences in these parameters between AAC group and sham group were significant by 3 weeks and reached peaks at 4th week. Following AAC, the plasma BNP was gradually elevated compared with the sham group at 3rd and 6th week. It was concluded that this modified AAC model can develop LVH, both stably and safely, by week four post-surgery; echocardiography is able to assess changes in chamber dimensions and systolic properties accurately in rats with LVH.

Alterations in cardiac structure and function in a modified rat model of myocardial hypertrophy / 华中科技大学学报（医学）（英德文版）

This study was aimed to establish a stable animal model of left ventricular hypertrophy (LVH) to provide theoretical and experimental basis for understanding the development of LVH. The abdominal aorta of male Wistar rats (80-100 g) was constricted to a diameter of 0.55 mm between the branches of the celiac and anterior mesenteric arteries. Echocardiography using a linear phased array probe was performed as well as pathological examination and plasma B-type natriuretic peptide (BNP) measurement at 3, 4 and 6 weeks after abdominal aortic constriction (AAC). The results showed that the acute mortality rate (within 24 h) of this modified rat model was 8%. Animals who underwent AAC demonstrated significantly increased interventricular septal (IVS), LV posterior wall (LVPWd), LV mass index (LVMI), cross-sectional area (CSA) of myocytes, and perivascular fibrosis; the ejection fraction (EF), fractional shortening (FS), and cardiac output (CO) were consistently lower at each time point after AAC. Notably, differences in these parameters between AAC group and sham group were significant by 3 weeks and reached peaks at 4th week. Following AAC, the plasma BNP was gradually elevated compared with the sham group at 3rd and 6th week. It was concluded that this modified AAC model can develop LVH, both stably and safely, by week four post-surgery; echocardiography is able to assess changes in chamber dimensions and systolic properties accurately in rats with LVH.

Case-control study on bone setting manipulation for the treatment of over degree II supination-eversion fractures of ankle joint / 中国骨伤

<p><b>OBJECTIVE</b>To explore therapeutic effects of bone setting manipulation for the treatment of over degree II supination-eversion fractures of ankle,and analyze manipulative reduction mechanism.</p><p><b>METHODS</b>From 2005 to 2008, 95 patients with over degree II supination-eversion fractures of ankle were treated respectively by manipulation and operation. There were 43 cases [11 males and 32 females with an average age of (44.95 +/- 12.65) years] in manipulation group, and 2 cases were degree II, 11 cases were degree III, and 30 cases were degree IV. There were 52 cases [21 males and 31 females with an average age of (39.96 +/- 13.28) years] in operative group,and 6 cases were degree II, 18 cases were degree III, and 28 cases were degree IV. Bone setting manipulation and hard splint external fixation were applied to manipulative group. Operative reduction internal fixation was performed in operative group. X-ray was used to evaluate reduction of fracture before and after treatment, 2 months after treatment. Ankle joint function was evaluated according to Olerud-Molander scoring system after 6 months treatment.</p><p><b>RESULTS</b>All patients were followed up with good reduction. Three cases occurred wound complication in operative group, but not in manipulative group. In manipulation group, 19 cases got excellent results, 20 cases good and 4 cases fair; while in operative group, 30 cases got excellent results, 20 cases good and 2 cases poor. There were no significant differences in fracture reduction and ankle joint function recovery between two groups (P > 0.05). Efficacy of operative treatment was better than that of manipulative treatment at degree IV fracture (P < 0.05).</p><p><b>CONCLUSION</b>Bone setting manipulation is a good method for treating supination-eversion ankle joint fractures, which has advantages of simple and safe operation, reliable efficacy. For ankle join fracture at degree IV, manipulative reduction should be adopted earlier, and operative treatment also necessary</p>

Screening and identification of polypeptides specifically binding to human B type natriuretic peptide from 12TM phage display peptide library / 南方医科大学学报

<p><b>OBJECTIVE</b>To identify and characterize the polypeptides specifically binding to human B type natriuretic peptide (BNP) screened from 12TM phage display peptide library.</p><p><b>METHODS</b>The BNP-binding peptides were screened from 12TM phage display peptide library and identified by ELISA.</p><p><b>RESULTS</b>After 4 rounds of screening, 10 of the 16 phage clones were identified as the positive clones which could bind to BNP. Five amino acid sequences were obtained in the 10 positive clones. Dose-dependent ELISA results demonstrated that the screened polypeptides could specifically bind to BNP.</p><p><b>CONCLUSION</b>These screened polypeptides can bind specifically to BNP, which provides a basis for further research on expression and purification of anti-BNP polypeptides and the development of the detection kit of BNP.</p>

Role of sphingosine 1-phosphate receptor signaling in hematopoietic stem/progenitor cell transmigration / 南方医科大学学报

<p><b>OBJECTIVE</b>To determine the role of sphingosine 1-phosphate receptor (S1PRs ) signaling in CD34+ hematopoietic stem/progenitor cell transmigration.</p><p><b>METHODS</b>CD34(+) cells were separated by Ficoll density gradient centrifugation and incubated in DMEM medium with 10% fetal calf serum. The cells were pretreated by FTY720, with or without pertussis toxin (PTX) and antiCXCR4 mAb in the medium, followed by addition of 100 ng/ml SDF-1 into the lower chamber of a Costar 24-well transwell. The migrated cells were counted using FACS and the migrating rates were determined. The expressions of sphingosine 1-phosphate receptors were analyzed in CD34(+) cells before and after the transmigration by reverse transcriptase- polymerase chain reaction (RT-PCR). Cord blood CD34(+) cells were treated with or without FTY720 (10(+) mol/L), and the expressions of CD49d (VLA-4), CD11a (LFA-1), and CD62L (L-selectin) were analyzed at 1, 8, and 16 h after the treatment.</p><p><b>RESULTS</b>While FTY720 did not affect spontaneous migration, a substantial increase of SDF-1-induced transmigration was observed in the presence of FTY720 (15.26 2.14 to 28.64 2.37). The FTY720-enhanced transmigration was completely blocked by addition of PTX or antiCXCR4 mAb. S1p1-5 was expressed in fresh isolated cord blood CD34(+) cells. The migrating cells stimulated by FTY720 and SDF-1 only expressed S1P1, S1P3, and S1P4. The expressions of CD49d, CD11a and CD62L on CD34(+) cells treated with FTY720 remained unchanged at the selected time points as compared with the control.</p><p><b>CONCLUSIONS</b>S1PRs are involved the transmigration of CD34(+) cells. The activation of S1PRs results in increased chemotactic response of CD34(+) to SDF-1. These effects are mediated through CXCR4 and PTX-sensitive Gi proteins. Only the CD34(+) cells expressing the specific receptors can rapidly transmigrate. The activation of the S1PRs does not affect the expressions of the adhesion molecules on cord blood CD34(+) cells.</p>

Taking ilium inner table to repair the articular surface defects of complex fracture of tibial plateau / 中国骨伤

<p><b>OBJECTIVE</b>To study the feasibility of application of ilium inner table to repair the articular surface defects of tibial plateau complex fractures.</p><p><b>METHODS</b>Twenty-three patients with tibial plateau complex fractures included 17 males and 6 females with an average age of 28.3 years old ranging from 18 to 51 years. The area of the articular surface defects ranged from 1 cmx2 cm to 3 cmx3 cm, averaged 6.7 cm2. Taking ilium inner table with periosteum after trimmed and implanting into the articular surface defect area with the concavity upward and drilled with diameter 1.5 mm Kirschner pin interval 3 to 4 mm. Bone grafting were placed under the ilium inner table and were fixed by T-shaped or L-shaped plate. The wounded limb were braked by plaster for 4 weeks after operation.</p><p><b>RESULTS</b>Twenty-three patients were followed-up for 8 months to 3 years, averaged 13.6 months. X-ray film showed solid union and the smooth articular surface in all cases. According to the Rasmussen evaluation system, the results were excellent in 11 cases, good in 8 cases,fair in 3 cases, poor in 1 case.</p><p><b>CONCLUSION</b>Taking ilium inner table to repair the articular surface defects of tibial plateau complex fractures is a good resolving measures. It can repaire major area of articular surface defects, restore the smooth articular surface and acquire good function of knee joint with easy to operate, less complications at donor area.</p>