Table of Content

05 October 2013, Volume 33 Issue 10

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A clinical and genetic study of a rare adrenoleukodystrophy kindred

2013, 33(10):  1223-1228. 

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Objective To analyse the clinical features of a kindred with adrenoleukodystrophy (ALD) and detect the mutation of ABCD1 gene. Methods A Chinese ALD kindred with four affected males in four-generation was studied. ALD was diagnosed according to clinical manifestations, cranial MRI image and serum very long chain fatty acid (VLCFA) levels. Mutation of ABCD1 was detected by direct DNA sequencing of polymerase chain reaction amplification product. Results Primary adrenocortical insufficiency and neurological dysfunction were the main manifestations of the patients. MRI image indicated extensive cerebral white matter demyelination. Serum VLCFA level was significantly high. A novel missense substitution (c.4037 C>T) in exon 8 of ABCD1 was identified. All affected males were hemizygotes and female carriers were heterzygotes. Conclusions The typical manifestations of ALD were primary adrenocortical insufficiency and neurological dysfunction. A missense substitution (c.4037 C>T) in exon 8 of ABCD1 was novel mutation firstly detected in this Chinese pedigree with ALD.

In Vitro synthesized modified mRNA can enter the hUC-MSCs

2013, 33(10):  1229-1234. 

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Objective To investigate the stability and efficiency of the in vitro synthesized modified mRNA, when it's transfected into the human umbilical cord mesenchymal stem cells（hUC-MSCs), so to establish a platform of using mRNA to induce the hUC-MSC differentiate into other cells for cell therapy. Methods To get the plasmid construct as the template for mRNA synthesis and synthesize the mRNA of eGFP. When the modified mRNA was transfected into the hUC-MSCs, using flow cytometry to analyze the transfect efficiency and determine the best transfect doses also to find the half-life time of mRNA. The same method was used to synthesize the mRNA of Pdx1 and to transfect it into the hUC-MSCs, then to measure the transfect efficiency by immunoflurescence. Result The in vitro synthesized modified mRNA of eGFP can enter the hUC-MSCs efficiently, the best doses for transfection is 1.5μg/mL and the mRNA can stay in the cell stably for 3 days. The in vitro modified mRNA of Pdx1 also can enter the hUC-MSCs. Conclusion In vitro synthesized modified mRNA has good stability and can enter the cells and translate into protein.

rHSG gene regulates in COPD rats,airway fibroblast proliferation and apoptosis

2013, 33(10):  1235-1241. 

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Objective Using liposome-mediated carry the eukaryotic expression vector of rats proliferation inhibition gene (rHSG) and transfected into chronic obstructive pulmonary disease (COPD) model of rat airway fibroblasts, in order to lay a basis of gene therapy that intervent the proliferation of COPD airway fibroblasts. Methods The recombinant plasmid pEGFP-rHSG were identified and sequenced.After transfected the recombinant plasmid pEGFP the-rHSG to COPD rat model of airway fibroblasts through cationic liposome and infected 24 hours, 48 hours, 3 days,5 days,7 days,detect the expression levels of rHSGmRNA on different times by real-time fluorescence quantitative polymerase chain reaction (Real time-PCR), finally,using MTT, flow cytometry detect the proliferation and apoptosis of transfected fibroblast cell.Results The recombinant plasmid pEGFP- rHSG constructed successfully.The expression of rHSGmRNA began to increase when transfected 24 hours，peak at 7 days,the differences in rHSGmRNA expression between rHSG meet each transfection group, P <0.05; After 48 hours，MTT assay show that the proliferation of airway fibroblast was significantly higher inhibited in rHSG gene transfection group than in model control group,statistically significant difference (P<0.05).After 48 hours,apoptosis were increasing by flow cytometry, compared with the model group, the difference was significant(P<0.05).Conclusion When exogenous rHSG gene transfected into the COPD rat airway fibroblasts, inhibit proliferation and induce its apoptosis.

Melatonin attenuates the change of hypothalamic free radicals and serum thyroxine induced by chronic social stress in rats

2013, 33(10):  1242-1246. 

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Objective To observe the effect of melatonin (MLT) on oxidative damage and neuroendocrine derived from chronic social stress (CSS). Methods Rats were classified into 3 groups: control group、CSS group and MLT group. CSS was induced by rats housed with aggressive male rats and female rats in a crowded cage for 8 days. MLT solution was administered 3 days before accepting CSS stimulation. Results SG and MG exhibited dramatic loss of body weight. Increased plasma corticosterone (CORT) and decreased serum tT3 (total triiodothyronine), tT4 (total thyroxine), fT3 (free triiodothyronine), fT4 (free thyroxine) and TSH (thyroid stimulating hormone) levels were evident in SG. The activity of glutathione peroxidase (GSH-PX) and content of reduced glutathione (GSH) in stress group (SG) decreased significantly, Melatonin group (MG) showed a marked increase in the activity of GSH-PX and decrease in the content of MDA. In addition, in open field test, scores of horizontal activity in central region in SG were significantly fewer than those in CG and MG. Conclusion The present study indicated that CSS could induce oxidative stress and impact on neuroendocrine, e.g. thyroid functions. But these changes could be partly reversed or attenuated by MLT.

Effect of C-KIT mutations on immunophenotype and proliferation of hematopoietic progenitors

2013, 33(10):  1247-1251. 

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Objective To investigate the molecular mechanism through which different C-KIT mutations influence the development of CBF-AML. Methods Assessment of exogenous KIT receptor and differentiation antigens on the surface of stably transfected EML cells was performed with flow cytometry; Cell proliferation in the presence or absence of IL-3 or Epo was analyzed using WST-1 cell proliferation assay; EpoR expression in different cell lines was identified with Western Blot assay. Results Human KIT receptor was expressed on the surface of transfected EML cells, Del417-419insY and D816V mutations led to a decrease in percentage of B220+ cells and an increase in Sca-1+ cells, albeit to a more extent for D816V mutation. Neither of them influenced positive percentage of CD34, Gr-1, Mac-1 and Ter119 in different cells. Del417-419insY mutant can cooperate with IL-3 or Epo to promote cell proliferation, while only IL-3 had a synergic effect with D816V mutant. Western Blot result confirmed the absence of EpoR expression in EML-D816V cells. Conclusion Distinct C-KIT mutations may differentially alter immunophenotype of hematopoietic progenitors and enhance cell proliferation, suggesting their relationship with development and prognosis of CBF-AML.

Effect of LY294002 and rapamycin on bim and bax in coxsackievirus B3 induced HeLa cells

2013, 33(10):  1252-1258. 

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Objective To observe the effect of the inhibitor of mTOR (mammalian target of rapamycin) Rapamycin and the inhibitor of PI3K (phosphatidylinositol kinase 3-kinase) LY294002 on Bim and Bax---members of pro-apoptosis factors in Bcl-2 family in CVB3 (coxsackievirus B3) induced HeLa cells, further investigate the roles of PI3K and mTOR signaling pathway playing in the CVB3 induced apoptosis. Methods To construct a cell model of VMC (viral myocarditis) with subculture HeLa cells. HeLa cells infected by CVB3 were treated with 10 nM Rapamycin and 25 μM LY294002 according to the cell toxicity test. The Bim and Bax expressions were determined by both RT-PCR and Western blot. Results Compared with controls (CVB3+DMSO), CPE (cytopathic effect) at 3h and 6h pi (postinfection) were no significant difference which at 12h and 24h pi, however, CPE in LY294002 and Rapamycin group were more obviously. Compared with Sham, the mRNA and protein expression of Bim decreased induced by CVB3 which blocked by LY294002 and Rapamycin compared with controls. The mRNA expression of Bax decreased early after CVB3 infection while protein expression increased gradually, which was dynamically affected by LY294002 and Rapamycin. Conclusion LY294002 and Rapamycin promote HeLa cells apoptosis and expression of both Bim and Bax with the infection time, which illustrates that PI3K and mTOR signaling pathway may play important roles in CVB3 induced VMC by regulating the Bim and Bax expressions.

Intermedin alleviates the development of pulmonary vascular structural remodeling induced by high pulmonary blood flow in rats

2013, 33(10):  1259-1263. 

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Objective To explore the regulating effect of intermedin (IMD) on the pulmonary vascular structural remodeling and pulmonary hypertension induced by high pulmonary blood flow in rats and its mechanism. Methods Twenty-one male SD rats were randomly divided into control group (n=7), shunt group (n=8) and shunt with IMD group (n=6) . Abdominal aorta and inferior vena cava shunting was produced in rats of the latter two groups. After 8 weeks, IMD [1.5 μg/(kg?h)] was administered into rats of shunt with IMD group subcutaneously by mini-osmotic pump for 2 weeks. Mean pulmonary artery pressure (mPAP) and the ratio of right ventricular mass to left ventricular plus septal mass [RV/(LV+SP)] was evaluated. The pulmonary vascular micro-morphologic changes of rats were observed. The content of NO in the serum and lung tissue homogenate was detected by nitric acid enzyme reduction method. The expression of eNOS protein in the lung tissue was detected by Western blotting analysis. Result Compared with those of control group, mPAP, RV/(LV+SP), the muscularization of small pulmonary vessels and relative medial thickness of pulmonary artery in rats of shunt group were all significantly increased. Meanwhile, the content of NO in the serum and pulmonary tissue homogenate and the expression of eNOS protein in rats of shunt group were all significantly decreased compared with those of control group. However, IMD significantly decreased the mPAP and RV/(LV+SP), alleviated the changes of pulmonary vascular micro-structure, with the elevation of the content of NO in the serum and pulmonary tissue homogenate and the expression of pulmonary eNOS protein in shunting rats. Conclusion IMD alleviates the development of pulmonary hypertension and pulmonary vascular structural remodeling induced by high pulmonary blood flow, through eNOS/NO path way.

Selenite induces mitochondrial damage and apoptosis in SW480 colorectal cancer cells

2013, 33(10):  1264-1268. 

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Objective We investigated the mechanism of apoptosis induced by selenite from the perspective of mitochondrial damage, aiming to provide evidence to support therapeutic role of selenite in colorectal cancer. Methods Divide SW480 into 2 groups，investigate the ROS level，apoptosis and mitochondrial damage induced by selenite via confocal microscope, flowcytometry (FCM) and so on. Using MnTMPyP inhibits ROS level to analysis the process, detcect the function of cytochrome C in the process. Results Selenite increases ROS level in SW480 cells, induces mitochondrial damage, decreases mitochondrial membrane potential, leading to mitochondria fragmentation, release of cytochrome C and apoptosis. Superoxide anion inhibitor MnTMPyP could inhibit the process. Conclusion Selenite can induce mitochondrial damage and apoptosis in SW480 via oxidative stress. Selenite has potential therapeutic effect in treatment of colorectal cancer.

MiR-33b down-regulation by DNA Methylation and its implications for gastric cancer

2013, 33(10):  1269-1274. 

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Objective To investigate the implications of miR-33b down-regulation by DNA Methylation in gastric cancer (GC) and the association between miR-33b level and various clinicopathological factors. Methods Expression of miR-33b was detected by Taqman real-time PCR in 42 paired GC samples. The association between miR-33b level and various clinicopathological factors was analyzed. Genomic DNA samples were amplificated by PCR after modified by sodium bisulfite using the EpiTect Bisulfite Kit then the methylation degree of CpG island upstream of miR-33b gene was detected by methylation specific PCR (MSP). Results The lower level expression of miR-33b was not associated with gender, age, venous invasion, position, borrmann typing, pT stage, pN stage, but significantly associated with gastric cancer metastasis (p<0.05) and regulated by CpG island hypermethylation (p<0.05). Conclusion miR-33b may be regulated by DNA methylation and act as a tumor suppressor in gastric cancer.

Expression variances of insulin signaling pathway-associated protein in the brain of newborn IUGR rats

2013, 33(10):  1275-1279. 

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Objective To investigate the expression variances of insulin signaling pathway-associated proteins and postsynaptic density protein (PSD95) in the brain of newborn IUGR rats, in order to explore the underlying developmental origin of neurodegeneration. Methods IUGR rat models were made by calorie restriction. RT-PCR and Western blot were applied to detect the level of insulin receptor (InsR), phospho-InsR, insulin receptor substrate -1(IRS-1) and PSD95 in the brain of newborn IUGR rats. SPSS 11.5 was applied to analyze the data. Results Compared with the control group, The body mass and the brain mass of newborn IUGR rats were both decreased（P<0.01）, while the ratio of brain mass/body mass was increased significantly（P<0.01）. In the brain of newborn IUGR rats, mRNA level of InsR was decreased（P<0.05）, protein levels of α（P<0.05）andβ（P<0.01）subunit of InsR and phospho-InsR（P<0.05） were decreased, IRS-1 was decreased（P<0.05）, and PSD95 was decreased too（P<0.01）. Conclusion There were expression variances of insulin signaling pathway-associated proteins and PSD95 in the brain of newborn IUGR rats, which might increase the liability of neurodegeneration in late years.

Effects of adiponectin on proliferation and oxidative stress level of INS-1 cultured with D-glucose of high concentration

2013, 33(10):  1280-1283. 

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Objective To investigate the effects of adiponectin(ADPN) on cell proliferation and the oxidative stress level of INS-1 cultured with high concentration of D-glucose and to illuminate the potential protection of ADPN on the INS-1.Methods The INS-1 cells culture in vitro ：The INS-1 cells were cultured with 5.5mmol/L glucose(control group), 25.6 mmol/L glucose(high glucose group), 25.6mmol/L glucose and concentration of 2.5mg/l adiponectin(ADPN group) , 25.6mmol/L glucose and concentration of 10.0μmol/L SB203580(SB203580 group) in random.After culture for 24,48,72h, the proliferation activity of INS-1 was observed by CCK-8.The content of MDA and T-AOC of the cell supernatant was measured by colorimetry. The expression of t-P38MAPK, p-P38MAPK were examined by Western blot.Results Compared with control group, INS-1 proliferation activity, the activation of P38MAPK, the content of T-AOC and MDA, have difference in high glucose group in 24,48,72h (P<0.05), and adiponectin or SB203580 could inhibit inhibit these changes(P<0.05). Conclusion Adiponectin could promote the proliferation of the INS-1,ameliorate the oxidative stress level and protect them in D-glucose of high concentration through the P38MAPK signaling pathway.

Wnt signaling takes part in down-regulation of hypoxia-induced fractalkine expression with LiCl in HUVECs

2013, 33(10):  1284-1287. 

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Objective To investigate the relationship between Wnt signaling pathway and hypoxia -induced fractalkine expression in human umbilical vein endothelial cells. Methods HUVECs were cultivated and divided into three groups randomly (control group, hypoxia/reoxygenation group, LiCl 10mmol/L group), which were used to establish the hypoxia/reoxygenation models. The mRNA expressions of fractalkine and GSK-3β were assessed by RT-PCR. The intracellular distribution and expression of fractalkine were determined by immunofluorescent staining. The expression of GSK-3β and β-catenin were analyzed with immunocytochemistry. Results Compared with control group, in hypoxia/reoxygenation group, the expression of GSK-3β and β-catenin were increased (P<0.05), the mRNA and protein expression of fractalkine increased remarkably (P<0.05). Compared with hypoxia/reoxygenation group, in LiCl groups, the mRNA and protein expression of fractalkine reduced significantly (P<0.05), the expression of GSK-3β reduced but β-catenin increased relativitily (P<0.05). However, the mRNA expression of GSK-3β had no statistically difference in all groups. Conclusion The hypoxia induced the damage of HUVECs through upregulating fractalkine expression. LiCl pretreatment inhibited fractalkine expression proportional. Its mechanism may be related to the activation of Wnt signaling pathway.

Respiratory syncytial virus induces the apoptosis of A549 associated with NF-κB signaling pathway activation

2013, 33(10):  1288-1292. 

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Objective To investigate the apoptosis mechanism of respiratory syncytial virus (RSV) and provide a new way of clinical therapy through the change of NF-κB after RSV infected A549 and IMR-90 cell. Methods RSV with the multiplicity of infection (MOI) 3 infecting A549 and IMR-90 cell, cell morphological change was observed using optical microscope, cell viability was detected with MTT, RSV viral titer was detected using empty spot test. Caspase-3, 8, 9 、cytochrome-c and NF-κB protein expression were detected using western - blot method. Results RSV infection A549 and IMR - 90 cell, A549 cell survival was lower than IMR - 90, intracellular virus titer was higher than IMR – 90 (P < 0.05). Cell showed obviously apoptosis change. Caspase-3, 8, 9 and cytochrome-c in time-dependence increased. Caspase-9 expressions were observed. In early (< 8h)RSV infection NF-κB was activated in time dependent manner, 8h to the peak, 24h began to decline. NF-κB expression was down-regulated after 36h RSV infection (P < 0.01). Conclusion RSV can through the endogenous and exogenous way caused A549 cell apoptosis. The endogenous way was primary. NF-κB played a certain role in the process.

Performance of bispectral index during epidural anesthesia and combined with propofol

2013, 33(10):  1293-1296. 

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Objective To evaluate the sedative effect of epidural anesthesia and interaction between epidural anesthesia and propofol using bispectral index (BIS). Methods 40 patients undergoing elective abdominal operation with general anesthesia participated in the study. After placing the epidural catheter, the patients were randomly allocated to 2 groups receiving either 5 mL of 1% lidocaine through the epidural catheter (group L) or 5 mL of epidural normal saline (group S), with 20 patients in each group. 8 minutes later, after confirmation of epidural administration, group L received 5～10 mL of combination of 1% lidocaine and 0.5% ropivacaine, whereas group S received 8 mL of epidural normal saline. BIS, MAP, HR and SpO2 were recorded before epidural injection, 8 min after first epidural injection, 15 min after first epidural injection and 30 min after first epidural injection. 30 min after first epidural injection, general anesthesia was induced with propofol via target-controlled infusion pump, the initial plasma concentration was 4 μg/mL. The effect site concentration of propofol when BIS firstly achieved 50 was recorded by another anesthesiologist who is blinded to the grouping. Following that, remifentanil and rocuronium were administrated to accomplish the endotracheal intubation. Follow-up was made for each patient postoperatively to identify possible intraoperative awareness. Results There were no significant changes in BIS after epidural infusion in both groups, and no significant differences exist between groups at all time points regarding BIS. The effect site concentration of propofol on BIS 50 in group L was significantly less than group S (P = 0.0157), 2.1 ± 0.5 μg/mL vs. 2.5 ± 0.5 μg/mL, respectively. Conclusion Epidural anesthesia alone doesn’t exert sedative effect measured by BIS. However, when combined with propofol, epidural anesthesia can potentiate the sedative effect of propofol.

Lipopolysaccharide Pre-treatment Increases Expression of HIF-1α in Cobalt chloride-induced murine macrophages

2013, 33(10):  1297-1301. 

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Objective To observe the effects of hypoxia simulated by cobalt chloride on the expression of hypoxia-inducible factor-1α(HIF-1α) and vascular endothelial growth factor(VEGF) in murine macrophage Raw264.7 induced beforehand by using LPS . Methods Raw264.7 were divided into control group，LPS（1mg/L）group，CoCl2（100μmol/L）group and LPS pre-treatment(LPS-Pre) group and incubated respectively in the culture containing corresponding reagent. The macrophages of LPS pre-treatment group were at first incubated for 8 hours in the culture with LPS of 1mg/L and then were transferred into the culture containing CoCl2 of 100μmol/L. HIF-1α and VEGF’ mRNA were detected by RT-PCR；protein of HIF-1α and VEGF were analyzed by Western blot and immunocytochemistry. Results Compared with those of control, mRNA and protein expression of both HIF-1α and VEGF can be up-regulated in CoCl2 group and LPS-Pre group（p<0.05）.Moreover, cells were pre-treated with LPS and followed activated by CoCl2, HIF-1α’s protein significantly higher（p<0.05）than that of induced only by CoCl2. Conclusion Pre-treatment with LPS can up-regulate HIF-1α and VEGF’s protein at the post-transcriptional level in macrophages under hypoxia environment.

The effects of migration of superparamagnetic iron oxide (SPIO)-Labeled mesenchymal stem cells by a magnet on brain infarction in rat model

2013, 33(10):  1302-1308. 

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Objective To observe the effect of rat bone marrow mesenchymal stem cells(BMSCs) labeled with superparamagnetic iron oxide(SPIO) in the magnetic field on the transplantation for rat focal cerebral ischemia. Methods The rat mesenchymal stem cells was labeled with SPIO in vitro , and prussian blue staining was used to check the rate of SPIO. The viability and proliferation activity of labeled stem cells was evaluated by trypan blue staining. The cck-8 assay was performed to detect the labeled stem cell proliferation activity in the magnetic field. The middle cerebral artery occlusion (MCAO) was established in a rat model, and the neurological severity score (NSS) was used to evaluate neurological function recovery of rats at definite time following the injection of transplanted cells. The transplanted BMSCs were analyzed in recipient rat brains by Prussian blue staining. The cerebral ischemia area was observed via TTC staining. The iron content of SPIO-labeled BMSCs was analyzed by atomic absorption spectrophotometer. Results Prussian blue staining showed that the rate of BMSCs labeled superparamagnetic iron oxide was close to 100％. Trypan blue staining indicated that there was no significant difference between labeled group and control group （P>0.05）. CCK-8 assay suggest that there was no significant difference between magnetic group and control group （P>0.05）. The BMSCs of ICSM group within the brain was significantly more than the BMSCs of ICS group (P < 0.05). The ICSM group with ischemia seven days later showed better compared to the other two groups. The difference was significant ( P< 0. 05) . The brain iron content in the ICSM group was largely more than that in the ICS group (P<0.05). Conclusion External magnetic field can target delivery bone marrow mesenchymal stem cells labeled with superparamagnetic iron oxide in vitro and in vivo, and can obviously improve the transplantation of SPIO-Labeled mesenchymal stem cells.

Selenite and Gossypol induce apoptosis in SW480 colorectal cancer cells

2013, 33(10):  1309-1313. 

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Objective We studied the cooperativity of selenite and gossypol to induce apoptosis in SW480 colorectal cancer cells. Methods Investigate the changes of Bcl-2 protein familys in SW480.SW480 cells were divided into three groups: 6μmol/L gossypol, 10 μmol/L selenite and both of them, then we observed the apoptosis by DAPI staining and fluorescence microscopy. Finally, we detected the cell death induced by different concentration gossypol and selenite in Bax/Bak deficient MEF cell. Results The expression of Bim and Bax proteins increased while Mcl-1 and Bcl-xl decreased in SW480 cells after selenite administration. Nucleus fragmentation rates and cell death in SW480 were significantly increased after the combined treatment of selenite and gossypol（P＜0.05）. In Bax/Bak deficient cells, gossypol can still promote apoptosis. Conclusion Selenite and gossypol can promote apoptosis via different pathways, selenite increases the sensitivity of tumor cells to apoptosis through Bim and Bax, while gossypol reduced the resistance to apoptosis of tumor cell, and the combination of them produced synergistic effect inducing apoptosis in SW480 cells.

Hemorrhoid rubber band ligation treatment in the patient using the CRH O’Regan Disposable Hemorrhoid Banding System

2013, 33(10):  1314-1316. 

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Objective To evaluate the efficacy, complications, success rate, and recurrence rate at 3 months for the CRH-O’Regan Disposable Hemorrhoid Banding System in the treatment of rubber band ligation （RBL）in outpatients. Methods 289 internal hemorrhoid grading included from grade I to IV outpatients were treated by RBL with CRH. Completion rate for surgery, intraoperative and postoperative complications and postoperative outcomes were statistically analyzed. Result All patients successfully completed the RBL with CRH, 245 cases (84.8%) were cured, 36 cases (12.4%) improvement in symptoms, 8 cases’ (2.8%)effects were not obvious, no patients’ symptoms had increased ; 2 cases (0.7%) had severe pain discomfort, 5 cases (1.7%) had postoperative dizziness; no acute complications, infection and urinary retention .Conclusion The RBL treatment of internal hemorrhoids with CRH has a good efficacy and safety.

Analysis of causes of death in 124 emergency cancer patients

2013, 33(10):  1317-1318. 

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Objective To analyze the causes of death in cancer patients. Methods In collection of 6316 cancer cases coming to the emergency clinic, the causes of death were analyzed in 124 cases. Results In the 124 cases, 25.8%（32/124）was due to the cancer growth itself while 74.2%（92/124）was resulted from complications. Most of the deaths were seen in patients with cancer of the lung, stomach and esophageal cancer. Cachexia，infection，respiratory failure, hemorrhages and shock were the causes of death frequently seen. The first two causes accounted for 25.8% and 22.6% of the total deaths in this series, respectively. Conclusion Complications are the major causes of death. The importance of their proper prevention needs to be emphasized.

The best point of Doppler guided hemorrhoidal artery ligation(DG-HAL)

2013, 33(10):  1319-1321. 

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Objective There are many treatments of II degree bleeding internal hemorrhoids ,one of them is Doppler guided hemorrhoidal artery ligation(DGHAL).We tried to find the best point of the ligation. Methods We chose 56 patients suffered with II degree Bleeding Internal Hemorrhoids and treated by Doppler guided hemorrhoidal artery ligation(DGHAL).Up to the ligation point ,3groups were made and . A 3-months follow-up was given and the efficacy was recorded. Results The middle ligation group had the least complication and the best efficacy significantly. Conclusion Doppler guided hemorrhoidal artery ligation(DGHAL)’s best point is about 2.0±0.5cm above the Dentate line

Decreased phosphorylation of eNOS at serine site in arteries of diabetic rats

2013, 33(10):  1322-1323. 

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Dynamic observation of peroxisome proliferator-activated receptorγ in process of experimental rats liver fibrosis

2013, 33(10):  1324-1325. 

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Isolation and identification of microvascular endothelial cells from rat spinal cord

2013, 33(10):  1326-1327. 

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The dual functions of P53 in the regulation of autophagy

2013, 33(10):  1328-1331. 

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P53, a critical suppression gene, has dual functions in the regulation of autophagy, depending on subcellular location, cytoplasm or nuclear. In the nucleus, P53 exerts its pro-autophagic function in transcription-dependent or independent manner. On the contrary, in the cytoplasm, P53 may inhibit the activation of autophagy, involving glycolysis metabolic transition. In conclusion, understanding its mechanisms will contribute to develop novel anti-cancer approaches.

Advances of collapsin response mediator protein 2 and its relation to neural diseases

2013, 33(10):  1332-1336. 

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Collapsin response mediator protein 2(CRMP2) plays a crucial role in neuronal development. It owns lots of post-translational modifications, of which phosphorylation modification is related to diseases closely. CRMP2 also has many biological functions. There is a close relationship between CRMP2 and neurologic disorders, but the mechanism is required to be elucidated. The researches in CRMP2 provided new strategies to prevent and cure the related diseases.

Progress in promoting osteogenesis of transplanted mesenchymal stem cells

2013, 33(10):  1337-1340. 

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Lower homing, surviving, proliferation, differentiation and osteogenesis of transplanted mesenchymal stem cells (MSCs) limit its use in bone disease. Recently, transplantation in combination with growth factors, prevascularized scaffolds, or regulation of signaling proteins and cytoprotective genes was applied to increase its therapeutic effects, and the results are encouraging.

The regulation of reactive oxygen species on cell apoptosis and proliferation

2013, 33(10):  1341-1344. 

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Reactive oxygen species ( ROS ) are byproduct of vivo aerobic cells in the metabolic process , including O2 - 、H2O2 、 ? OH and HO2- . Existing studies have shown that when the balance of ROS generation and clear broken , then ROS through damage mitochondria 、activate the death receptor , affect the expression of cytokines and activate the transcription factor , to participate and influence the cell signal transduction mechanisms , which has a crucial regulatory for the cell apoptosis and proliferation.

The antitumor activity of aspirin via COX-2-independent pathways

2013, 33(10):  1345-1347. 

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Recently, aspirin (ASA) has been found to be of antineoplastic activity. Apart from the classical pathway of inhibiting cyclooxygenase-2 (COX-2), there are major COX-2-independent pathways governing the anticancer mechanisms of ASA, including induction of nitric oxide synthase (NOS), regulation of NF-κB pathway and the Bcl-2 family, together with inhibition of Akt, mTOR and ERK activation. Of note, ASA’s antitumor activity varies in different types of cancer cells. In the same cancer cell, ASA triggers apoptosis via different pathway induced by distinct factors. Overall, the anticancer effects of ASA are diversified and cell-type specific.

Associated genes research of GWAS in type 2 diabetes mellitus

2013, 33(10):  1348-1351. 

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T2DM is a complex multi-gene disease,and 65 genes have been found whose gene variations were associated with T2DM with the help of GWAS in recent years. These genes play important roles in the development and function of pancreatic β cells, and also in peripheral insulin resistance. The find of associated genes with T2DM in GWAS provides new ideas for the intervention and treatment of T2DM.

The development of adenosine augmentation therapies (AATs) in epilepsy therapy

2013, 33(10):  1352-1355. 

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Adenosine is an endogenous anticonvulsant transmitter in the brain. Recent researches suggest increasing site- and event-specific adenosine (adenosine augmentation therapies AATs) may become a novel therapy for epilepsy. Systemic pharmacological AATs are effective in seizure suppression, however accompanied by wide-spread side-effects. Therapeutic tools to achieve focal AAT, stem cells have been engineered to release therapeutic amounts of adenosine using RNA interference or gene targeting technology, providing robust protection from spontaneous seizures, but determining the most effective and compatible strategies with clinical applications needs more future research.

The application of telemedicine in surgery and surgical education

2013, 33(10):  1356-1359. 

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Telemedicine provides a new way for medical students and medical professionals to acquire medical knowledge and have academic communication and clinical cooperation .This article reviews telemedicine and other surgery related innovations that benefit from developments in the field of telecommunication, and its application in surgery and surgical education.

Application of supervisor responsibility system in standardized training for residency of general practitioners

2013, 33(10):  1360-1362. 

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It is a new idea to adapted supervisor responsibility system in standardized training for residency of general practitioners. According to the disadvantages in the general medicine residency training and teachers quantity not sufficient, the supervisor responsibility system is adapted and used in the training, which is conformable for the training, and has its special characteristics. To some extent, it overcome the disadvantages of the traditional teaching methods, and alleviate the problem of shortage of teachers numbers. It has played a positive role in improving the students' research capacity and innovation ability.