Objective To investigate the inhibition effects of simvastatin on proliferation and migration of cultured porcine coronary vascular smooth muscle cells (SMCs) induced by oxidized lower density lipoprotein (OX-LDL). Methods Porcine coronary SMCs were incubated with serum-free medium containing different concentrations of simvastatin. Infiltration of ~3H-TdR into SMC DNA was measured as an index of cell proliferation. Simvastatin's effect on migration was tested with Sarkar's technique. Results Different concentrations of simvastatin (10~(-8)-10~(-5) mol/L) caused a decrease in ~3H-TdR infiltration in porcine coronary SMC. The migration number of porcine coronary SMCs also decreased proportional to the simvastatin concentration. Conclusion Simvastatin can inhibit the proliferation and migration of porcine coronary SMCs stimulated by OX-LDL in a dose-dependent manner.

Objective To study the effectiveness of different treatment modalities for iatrogenic injury of distal common bile duct during operation.Methods We browsed Chinese Medical Full-text Data-base with the term of “distal common bile duct injury”.All the clinical studies associated with perioperative latrogenic injury of distal common bile duct and adjacent tissue published after 1990 were enrolled,and we collected the clinical data,mortality and reoperation rate with different treatments for analysis.Results Thirty-four case series and case reports with 233 patients were included.14 patients with isolated duodenal injury were excluded.The overall mortality of the remaining 219 patients was 9.6％,and the reoperation rate was 17.4％.A total of 145 patients who were diagnosed with distal common bile duct injury during and after operation from 21 articles were compared.The mortality and reoperation rate were both 1.9％ among 106 patients who were diagnosed during operation.The figures were 43.6％,and 84.6％ among 39 patients who were diagnosed after operation,respectively.In 9 articles with 46 patients,the clinical outcomes of 21 patients who were treated by intraoperative suture was compared with 25 patients who underwent enhanced biliary and retroperitoneal drainage.The mortality and reoperation rates were 0 in both groups.Conclusions Early detection and management are crucial to perioperative common bile duct injury.Furthermore,no significant difference of clinical outcomes observed between bile drainage and perforation suture groups.

Objective To evaluate gastrobiliary duct drainage in the treatment for iatrogenic distal common bile duct injury found during the operation.Methods We analyzed clinical data of 17 cases with application of gastrobiliary duct drainage in immediate treatment for the injury of distal common bile duct found during the operation from June 2010 to June 2016.Postoperative bile drainage,postoperative gastrointestinal function recovery,time for removal of the gastrobiliary duct and hospitalization time were recorded.Postoperative bile leakage,intestinal fistula and pancreatic leakage were observed.Patients were followed up until June 2016.Results The mean volume of bile drainage on the third postoperative day were (310 ± 112)ml,the mean time of postoperative gastrointestinal function recovery were (3.0 ± 1.5) days,time for removal of the gastrobiliary stent were (7.5 ± 1.0) days and hospitalization time were (9.5 ± 1.5) days.There was no postoperative bile leakage,intestinal fistula and pancreatic leakage.All patients were followed up for a median time of 12 months (range,1-45 months).Meanwhile,we found no significant biliary strictures and cholangitis patients.Conclusion Gastrobiliary duct drainage is a simple,rational and effective treatment for iatrogenic injury of distal common bile duct during common bile duct exploration.

Objective: To evaluate the efficacy and safety of tirofiban infusion to infarct related vessels on patients with ST segment elevation myocardial infarction (STEMI) during emergency percutaneous coronary intervention (PCI). Methods: From Jan 2013 to Jun 2014, a total of 30 STEMI patients were enrolled as tirofiban group (tirofiban 500μg was infused to infarct related vessels during emergency PCI), and received intravenous drip of tirofiban 0.1 μg•kg-1•min-1 for 24h after stent implantation; another 31 STEMI patients were regarded as pure stenting group during the same period and they received direct stent implantation during emergency PCI. Computer-assisted Quantitative Blush Evaluator (QuBE) score, left ventricular ejection fraction (LVEF) during hospitalization and after six-month follow-up and incidence rate of major adverse cardiovascular events were compared and analyzed between two groups. Results: There were no significant difference in baseline data between two groups, P>0.05. Compared with pure stenting group, after six months, there were significant rise in QuBE score [(10.88±5.03) scores vs. (14.70±6.69) scores] and LVEF [(57.19±4.59)% vs. (59.80±5.34)%], and significant reduction in incidence rate of MACE (35.5% vs. 10.0%) in tirofiban group, P<0.05 all. Conclusion: Tirofiban application in infarct related vessels during emergency PCI in STEMI patients can effectively and safely improve myocardial microcirculation perfusion level and it is worth extending.

BACKGROUND:Bone marrow derived mesenchymal stem cells (BMMSCs) have multi-differentiation potentials, possessing a repairing capability for the sectional bone defects if combined with degradable porous β-tricalcium phosphate china. This provides a new idea in clinical repair of various bone defects.OBJECTIVE: To explore in radiology the curative effect of implanting porous β-tricalcium phosphate and autogenous BMMSCs compound to treat bone defects.DESIGN: A randomized controlled study with callus growth at various healing periods as subjects for observation. SETTING: Department of Orthopaedics, Renmin Hospital, Wuhan University; Tissue Engineering Center, Research Institute of Basic Medicine,Academy of Military Medical Sciences of Chinese PLAMATERIALS: This experiment was carried out at the Tissue Engineering Center, Research Institute of Basic Medicine, Academy of Military Medical Sciences of Chinese PLA between March and September 2002. Twenty China healthy adult sheep were randomized into the groups of blank control group (4 sheep), simple implantation group (8 sheep) and complex implantation group (8 sheep).METHODS: Under general anesthesia and aseptic condition, 10-15 mL of sheep marrow was extracted; MSCs were separated and cultured before combined with porous β-tricalcium phosphate china for tissue engineering bone construction. Rats in each group were cut off 21mm long metatarsus in the middle section of metatarsus bonestem. Β-tricalcium phosphate china and autogenous MSC compound was implanted into the sheep of the complex implantation group; β-tricalcium phosphate china was implanted into the simple implantation group; and the bone defects in the blank control group remained untouched. Then the incision was sutured.X-ray filming was carried out right after the operation, as well as 1, 3,and 6 months after the operation for radiological appraisal (scored for1 if bone union formed in one surface of bone defect, but scored 0 if no boneunion formed in any surface of bone defect, and scored 4 if bone union formed in front, back, lateral surfaces and the center of bone defect), Xray radiation-resisting density was analyzed to compare the results of bone defect repair.MAIN OUTCOME MEASURES: The postoperative general condition and general observation, as well as the results of the radiological analysis of the bone defects of all sheep.RESULTS: Totally 20 sheep were brought into this xperiment and all entered the stage of result analysis. ① The postoperative general condition:Sheep regained consciousness 2-6 hours after the operation without incision infection and loosing of internal fixation. Their spirit gradually was back to normal 1 week after the operation, at which time the injured legs could touch ground but were incapable of bearing load, and the affected legs could bear load 2 weeks after the operation, walked slightly lamely 3 weeks after the operation, and even moved freely without limp 4 weeks after the operation. ②The general condition 6 months after the operation: In pure implantation group, the surface white hyaline cartilage-like tissues were gradually calcified, with both ends connected with the host bone by bone bridge, but china granules could still be easily observed; while no implantation substance could be observed in compound implantation group,with the boundary between implantation substance and host bone vanished,and bone defect became basically the same as host bone. However there was no bone tissue formed in bone defect at various postoperative time points in the blank control group. ③ Radiological analysis of the bone defects at various postoperative time points: The radiological rating score was obviously higher in complex implantation group at the time poin ts of 3, 6 months after the operation compared with the pure implantation group [(2.3±0.3), (1.8±0.5); (3.3±0.5), (2.6±0.6), P ＜ 0.05]. ④ Radiological analysis of bone callus thickness and the relative value of radiation-resisting density at various postoperative time points: The bone callus thickness in the complex implantation group was obviously lower than that of the pure implantation group at the postoperative time point of 6 months (4.62 vs 7.64, P ＜ 0.05), with relative value of radiation-resisting density obviously higher than that of the pure implantation group (70.4±1.5 vs 61.18±1.2, P ＜ 0.05).CONCLUSION: Radiological appraisal and bone defect density measurement can well reflect the dynamical repairing process of bone defects; the implantation of porous β-tricalcium phosphate china and autogenous BMMSCs compound into sheep can enhance the repair of large sectional bone defect.

Objective To investigate the relationship between CRP, plasminogen activator inhibitor type 1 (PAI-1) levels, PAI-1 gene promoter 4G/5G polymorphism and the type of acute myocardial infarction (ST elevation myocardial infarction, STEMI vs the non-ST elevation Myocardial infarction, NSTEMI). Methods One hundred seventy-six consecutive patients with AMI were included for the study, of whom 60 had STEMI and 56 had NSTEMI, and 60 adults without cardiovascular and cerebrovascular disease were selected as controls. Blood samples were obtained from patients within 6 h of AMI and the plasma PAI-1, CRP, and the gene polymorphism were measured. Results Plasma levels of PAI-1 and CRP were higher in AMI groups, compared those in the control group, and plasma levels of PAI-1 were significantly higher in patients with STEMI compared to those with NSTEMI (80.12ng/ml VS.73.01ng/ml, P <0.01), while CRP levels were not significantly different between patient with STEMI and NSTEMI (3.87±0.79 mg/ml VS.4.01±0.69mg/ml, P>0.05). PAI-1 levels presented a significant correlation with CRP levels in the NSTEMI subjects. However, PAI-1 and CRP levels could explain the lack of a significant relationship between them in control and STEMI subjects.The frequencies of 4G/4G genotype in the AMI group were higher than those in the control group and higher in patient with STEMI than in patient with NSTEMI. Plasma levels of PAI-1 in subjects with 4G/4G genotype were significantly increased as compared to those in subjects with 4G/5G and 5G/5G genotype. Conclusions Plasma PAI-1 levels were associated with different myocardial infarction type, and PAI-1 promoter 4G/5G polymorphisms and CRP may be related to plasma PAI-1 levels.

Objective To establish a key technological system for spider fibroin gene code tandem connection , vector construction , prokaryotic expression and purification using genetic engineering in order to achieve MaSp 1 heterologous ex-pression in Escherichia coli and its separation and purification .Methods Isocaudarner ligation method was used to connect synthetic spider fibroin gene monomer code in tandem , and a recombinant clone concatemer was obtained .The identified recombinant clones were connected with prokaryotic expression vector pET 28a(+), and then transformed into E.coli BL21 (DE3).After being induced by IPTG for 6 hours, the expression product was identified by SDS-PAGE and Western blot-ting.Engineering bacteria were fermented in high density , and the obtained protein was purified through ammonium sulfate fractionation.Results and Conclusion The expression plasmids of MaSp1concatemers were successfully constructed , and the induced expression genetic engineering MaSp 1 protein was of the expected relative molecular mass .In addition, the pu-rity of the purified protein was above 80%.This study has developed crucial technologies for mass production of genetic en-gineering spider silk proteins .

Objective To explore the sleep characteristics of submariners during a long-term voyage, so as to provide scientific evidence for ensuring submariners with good sleep during long-term voyages. Methods The sleep status of submariners who participated in a long-term voyage was tested by Self-Rating Scale of Sleep (SRSS) before the voyage, and before and after each voyage section during the voyage. The sleep status variation of submariners who performed different types of tasks, from the beginning to the end of each voyage section and of each resting-on-the-sea section was analyzed respectively. Comparison of sleep scores was performed between submariners and surface ship crew in the second voyage section. Numbers of submariners with sleep problem were compared in each voyage section. Results Generally speaking, submariners' sleep status at the end of voyage section was significantly worse than that at the beginning of voyage section and that before the whole voyage (P<0.001, P<0.01), and the sleep status at the beginning of the third voyage section was significantly worse than that before the whole voyage (P<0.05). Submariners had a steady sleep status when taking a resting-on-the-sea before starting their first voyage section, which was no significant difference from that before the whole voyage (P>0.05). After finishing a voyage section and taking a resting-on-the-sea, submariners' sleep status returned to the level of pre-voyage (P>0.05), and was significantly better than that before the resting-on-the-sea (P<0.05, P<0.01). After finishing two voyage sections and then taking a resting-on-the-sea, the submariners' sleep status showed no obvious variation (P>0.05). Compared with that of surface ship crew who accomplished the same voyage section, submariners had an obviously better sleep status after taking a resting-on-the-sea (P<0.05). Meanwhile, submariners who finished a voyage section showed a significantly worse sleep status than those resting on the sea (P<0.01) and surface ship crew who finished a same voyage section (P<0.05). In each voyage section, submariners with sleep problems who finished resting-on-the-sea were significantly less than those who finished navigation (P<0.001, P<0.05). There was no significant difference in the number of submariners with sleep problems between those who taking non-resting and taking resting-at-dock after finishing the first voyage section (P>0.05), but the latter was significantly more than the former when the second voyage section was finished (P<0.05). During the resting-on-the-sea period, the numbers of submariners with sleep problems in both the second and the third voyage section were significantly more than those in the first voyage section (P<0.05, P<0.01). The numbers of submariners with sleep problems who implemented the third voyage section were significantly more than those who implemented the first and the second voyage section (P<0.01). Conclusions Generally, the sleep quality of submariners is significantly worse after accomplished a voyage section task, and the degree of sleep problems may be accumulated to worse and worse along with the increase of long-term voyage time. Whereas, submariners may have a significantly better sleep status after taking a resting-on-the-sea, implying that resting-on-the-sea is an effective way to ensure submariners a good sleep during a long-term voyage.

OBJECTIVE: To investigate the effect of hypoxia on cell viability and the endothelial differentiation potential in human umbilical cord derived mesenchymal stem cells (UC-MSCs), and to assess the in vitro protective role of VEGF under low oxygen tension. METHODS: MSCs were isolated from human umbilical cords and cultured in vitro. The morphological and phenotypic characterizations of human UC-MSCs were analyzed. The hypoxia induction was performed with or without the presence of 50 ng/mL of VEGF for different lengths of time. The cell proliferation, apoptosis, and reactive oxygen species (ROS) generation were assessed. Meanwhile, the endothelial differentiation potential of the UC-MSCs was measured. RESULTS: An increased apoptosis and ROS generation but reduced proliferation rate were observed at early stages (6, 12 h) after transferring the UC-MSCs from the atmospheric condition to the hypoxia condition. However, the UC-MSCs presented equal proliferation and apoptosis levels under hypoxic condition as compared with those under the atmospheric condition at the later stages (24, 72 h). A high concentration of exogenous VEGF (50 ng/mL) attenuated the increased apoptosis and inhibited the proliferation of UC-MSCs, induced by a short-term hypoxia treatment. After 14 days of exogenous VEGF induction under the hypoxia condition, the UC-MSCs acquired an early endothelial phenotype consisting of a mature endothelial molecule and some endothelial functions. CONCLUSION UC-MSCs progressively adapt to hypoxia in a step-by-step manner and maintain differentiation potential under hypoxia condition. VEGF can protect the UC-MSCs from cell damage and induce a differentiation of UC-MSCs toward endothelial lineage under hypoxic conditions.
Apoptosis ; drug effects ; Cell Differentiation ; drug effects ; Cell Hypoxia ; Cell Proliferation ; Cell Survival ; Cells, Cultured ; Humans ; Mesenchymal Stem Cells ; cytology ; Protective Agents ; pharmacology ; Reactive Oxygen Species ; metabolism ; Umbilical Cord ; cytology ; Vascular Endothelial Growth Factor A ; pharmacology

Objective:To explore the effect of estrogen-related receptor α (ERRα) on lipopolysaccharide (LPS)-induced vascular endothelial cell apoptosis and tight junction protein degradation.Methods:RPMVECs transfected with shERRα were cultured in vitro and divided into four groups: Normal control group (Ctr group); shERRα knockdown group (shERRα group); normal cells + LPS treated group (LPS group): The cells in the six-well plates were cultured in serum-free medium for 12 h, and then treated with 20 μg/mL LPS for 12 h; and shERRα+LPS group: ERRα knockdown cells were treated as the LPS group. ROS fluorescence kit was used to detect the intracellular ROS levels . Apoptosis ratio was detected by TUNEL staining, AnnexinV-FITC and PI. Cell membrane ZO-1 expression was detected by cellular immunofluorescence, and the levels of apoptosis-related proteins Bcl-2, Bax, Smac, Cytochrome c, and tight junction protein ZO-1, as well as the expression of Occludin, JAM-A and E-Ca at molecular level were detected by Western blot.Results:Compared with the Ctr group and the shERRα group, the ROS level, apoptosis rate (TUNEL test: 16.44 ± 2.55; and flow cytometry test: 23.56 ± 2.22), the expression of pro-apoptotic proteins Bax, Smac and Cytochrome c were increased in the LPS group, while the expression of anti-apoptotic proteins Bcl-2 and tight junction protein were decreased. In the LPS group. Cellular immunofluorescence results showed that the ZO-1 was degraded in the cell membrane and the network structure was broken. Compared with the LPS group, inhibition of ERRα in the shERRα+LPS group increased cell damage.Conclusions:ERRα can negatively regulate the apoptosis and affect the function of pulmonary microvascular endothelial cells, thereby regulating sepsis-induced acute lung injury.