Sphingosine-1-phosphate in mitochondrial function and metabolic diseases.

Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid metabolite. The past decade has witnessed exponential growth in the field of S1P research, partly attributed to drugs targeting its receptors or kinases. Accumulating evidence indicates that changes in the S1P axis (i.e., S1P production, transport, and receptors) may modify metabolism and eventually mediate metabolic diseases. Dysfunction of the mitochondria on a master monitor of cellular metabolism is considered the leading cause of metabolic diseases, with aberrations typically induced by abnormal biogenesis, respiratory chain complex disorders, reactive oxygen species overproduction, calcium deposition, and mitophagy impairment. Accordingly, we discuss decades of investigation into changes in the S1P axis and how it controls mitochondrial function. Furthermore, we summarize recent scientific advances in disorders associated with the S1P axis and their involvement in the pathogenesis of metabolic diseases in humans, including type 2 diabetes mellitus and cardiovascular disease, from the perspective of mitochondrial function. Finally, we review potential challenges and prospects for S1P axis application to the regulation of mitochondrial function and metabolic diseases; these data may provide theoretical guidance for the treatment of metabolic diseases.

[Correlation of blood flow assessed by CT perfusion imaging and microvascular ultrastructure in non-small cell lung cancer: a preliminary study].

OBJECTIVE: To investigate the correlation between blood flow assessed by CT perfusion imaging and characteristics of microvascular ultrastructure in non-small cell lung cancer (NSCLC). METHODS: twenty-eight patients with non-small cell lung cancer proven surgically and pathologically underwent perfusion CT examination. The patients were divided into a hyper-perfusion group and a hypo-perfusion group by the median value of blood flow, and then the differences of microvascular ultrastructure in the two groups were analyzed. RESULTS: The median BF value of the 28 patients was 36.40 ml×100 g(-1)×min(-1). Take this median value as the boundary, the group with hypo-perfusion showed a significantly lower BF value than the group with hyper-perfusion [(30.84 ± 4.79) ml×100 g(-1)×min(-1) vs. (49.67 ± 10.89) ml×100 g(-1)×min(-1), t = -5.925, P < 0.001]. The group with lymph node metastasis showed a significantly lower BF value than the group without lymph node metastasis [(30.78 ± 5.24) ml×100 g(-1)×min(-1) vs. (50.73 ± 11.16) ml×100 g(-1)×min(-1), t = 3.490, P = 0.015]. The maturity of microvessels of the hyper-perfusion group was higher than that of the hypo-perfusion group. Under the electron microscope, the microvessels in the hypo-perfusion group showed a more narrow lumen, poorer integrity of basement membrane, a more close relationship between cancer cells and microvascular wall, and cancer cells were more easily seen in the microvascular lumen. CONCLUSION: The blood flow value of CT perfusion imaging may be related with the abnormal microvascular ultrastructure, and may be helpful to the prediction of metastasis risk in NSCLC.

[In vitro culture of Toxoplasma gondii tachyzoites in HFF and HeLa cells].

OBJECTIVE: To study the proliferation of Toxoplasma gondii RH strain tachyzoites in human foreskin fibroblast (HFF) cells and HeLa cells. METHODS: HFF cells and HeLa cells were cultured in 35 mm cell culture dishes with glass cover slip. Confluent cells were co-cultured with tachyzoites which purified by 3 microm filter membrane. At 0.5, 1, 2, 4, 8, 12, 24, 36, 48, 72, and 96 h after co-culture, the invasion of tachyzoites into the cells and proliferation in cells were observed with Giemsa staining. RESULTS: In 4 h after co-culture, there were dozens of T. gondii tachyzoites in the HFF cells. At 24 h many pseudocysts emerged. At 72 h most of the cells were destroyed by tachyzoites. While cultured in HeLa cells for 8 h, there were only 3-5 tachyzoites in one cell, and pseudocysts emerged at 48 h. At 96 h after co-culture, most cells were destroyed. CONCLUSION: Toxoplasma gondii tachyzoites can be cultured in HFF cells and HeLa cells. The proliferation of tachyzoites in HFF cells was quicker than that in HeLa cells.

[Clinical study of lung mucoepidermoid tumors].

OBJECTIVE: To investigate clinical and pathological characteristics of lung mucoepidermoid tumors and summarize methods for diagnosing and treating it. METHODS: Records of a total of 2,751 consecutive patients with lung cancer were reviewed and 10 of whom with mucoepidermoid tumors were identified. Chest radiographs, computer tomographs, and bronchoscopes were performed to all of them. Eight of them underwent thoracotomy and some also received chemotherapy and/or radiotherapy. One patient received Chinese herb therapy only while another did not receive any therapy. RESULTS: Pathological examination showed that the 10 patients had low-grade mucoepidermoid tumors. All patients were alive. Seven patients who received thoracotomy did not recur. One patient whose tumor was located in trachea recurred 5.2 years after the operation. CONCLUSION: Bronchoscope can play an important role in the process of diagnosis. Pathological staging is vital for prognosis. Lung mucoepidermoid tumors should be treated with complete surgical resection with lymph node sampling and dissection and close follow-up.

[Effect of HOE642 on cardiac myocyte apoptosis in rat non heart-beating donors].

OBJECTIVES: To investigate the effect of HOE642 on cardiac myocyte apoptosis of the heterotopic heart transplantation of rat non heart-beating donors. METHODS: Totally 112 male Sprague-Dawley rats were randomly divided into 7 groups (n=16 in each group) C, the control group (normal hearts); S10, S30, and S45 (groups of transplanted hearts after 10, 30, and 45 minutes of asystole); and SH10, SH30, and SH45 (groups of transplanted hearts after 10, 30, and 45 minutes of asystole and infused with HOE642). After rata in the experimental groups were killed by warm ischemia the donators of the S10, S30 and S45 groups were infused with 5TH-1 for 30 minutes, and the dead rats in group SH10, SH30, and SH4 were infused with STH-1 and HOE642 (20 micromol/L) for 30 minutes. Heterotopic heart transplantation were processed by the method of neck Cuff. The heart specimens of S10, SH10, S30, and SH30 groups were taken after 48 hours of transplantation, and the heart specimens of S45 and SH45 groups were taken immediately after transplantation. Then apoptotic myocytes were detected with terminal deoxynucleotide transferase-mediated deoxyuridine-biotin nick end labeling method and the expressions of Bcl-2, Bax, and Caspase-3 proteins were detected by immunohistochemistry. RESULTS: The rats were discerned death when cardiac electric wave vanished after 9-11 minutes of bloodletting by transsection of abdominal aorta. The number of positive cardiac muscle cells in S10 and S30 groups were significantly larger than those in group SH10 and SH30 (P < 0.05). The levels of Bcl-2 protein expression in S10 and S30 groups were significantly lower than those in SH10 and SH30 groups (P < 0.05). The levels of Bax and Caspase-3 protein expression were significantly higher than those in SH10 and SH30 groups (P < 0.05). CONCLUSIONS: The rat model of a heterotopic heart transplantation on the cervical part is a convenient animal model for cardiac muscle protection. HOE642 can suppress rat cardiac muscle cells apoptosis (within 30 min) after death caused by warm ischemia.

[Effect of cariporide on the expression of bcl-2 and bax genes after neck heart transplantation from non heart-beating rats caused by warm ischemia].

OBJECTIVE: To detect the expression of bcl-2 and bax genes after heterotopic heart transplantation in rats that died of warm ischemia, and to explore the effect of cariporide on the protection of the ratos non heart-beating donors. METHODS: One hundred and twelve clearing Sprague-Dawley male rats were divided into 7 groups at random (each group contained 16 rats): the control group (Group C), the groups of transplanted hearts after 10, 30, and 45 min of asystolia (Group S10,S30,and S45), and the groups of transplanted hearts after 10,30, and 45 min of asystolia and infused with cariporide(Group SH10,SH30, and SH45).The experimental groups were sacrificed totally by warm ischemia, and heterotopic heart transplantation was processed by the Cuff method. The heart samples of S10,SH10,S30, and SH30 groups were taken at 48 hours after the transplantation, and the heart samples of S45, and SH45 groups were taken just after transplantation. The expression of bcl-2 and bax genes were detected by RT-PCR. RESULTS: The death of rats was affirmed when cardiac electric waves vanished after 9~11 minutes of transsection of abdominal aorta. On the RT-PCR test, the expression of bcl-2 gene was the highest and ROD value was maximum in the control group. The expression of bax gene was the lowest and ROD value was minimum in the control group. The ROD value of bcl-2 genes in S10 and S30 groups was less than that in SH10 and SH30 group. The ROD value was just the opposite, and there was stastistical difference (P<0.05).There was no statistical difference between Group S45 and Group SH45 (P>0.05). CONCLUSION: The model of heteroto-pic neck heart transplantation is a convenient animal model for the cardiac muscle protection. Cariporide can suppress the apoptosis of cardiac muscle cells in rats (within 30 min) after death caused by warm ischemia.

[Protection of cadaver lungs of non-heart-beating donor lung transplantation in rats].

OBJECTIVE: To investigate the methods to alleviate lung injury of non-heart-beating donor to attain better structure and function. METHODS: Sixty-four Sprague-Dawley rats were randomly divided into 4 groupsú a heart-beating donor group(HBD group), a non-heart-beating donor group without protective measures(NHBD-N group), a non-heart-beating donor group with continuous mechanical ventilation or with topical cooling on cadaver lung (NHBD-V group and NHBD-I group). After the transplantation, lung compliance,pulmonary function,wet/dry ratio of lung and content of energy metabolism were compared among the 4 groups. RESULTS: Due to the longer warm ischemic period, NHBD lungs suffered more injuries than HBD lungs. However, compared with NHBD-N group, the wet/dry ratio of the lung in NHBD-V group and NHBD-I group was lower(5.28+/-1.24,4.21+/-0.85,4.14+/-1.33,P<0.01),the lung injury index (14.35+/-3.21,11.28+/-3.26,10.41+/-2.85, P<0.01)and the count of white blood cells(425.60+/-86.47,316.30+/-56.24,295.50+/-70.26, P<0.01) were milder, while the lung compliance and preservation of energetic metabolte were better in the NHBD-V group and the NHBD-I group. CONCLUSION: Continuous mechanical ventilation or topical cooling may protect the NHBD lung during the warm ischemic period.

[Pneumonectomy for 210 cases of lung cancer].

OBJECTIVE: To summarize 210 cases of lung cancer that underwent pneumonectomy,and to study the therapy value of pneumonectomy for lung cancer. METHODS: Between January 1993 and December 2004,210 patients (169 males and 41 females) with lung cancer underwent pneumonectomy in our hospital with the mean age of 54.77 years. Out of the 210 cases 144 had left pneumonectomy, and 66 had right pneumonectomy. Among them, 26 experienced intrapericardial pneumonectomy,1 inferior vena cava reconstruction, and 1 carina reconstruction. All bronchial stumps were closed by manual suture,and systemic node dissection was performed routinely. RESULTS: No one died during the operation, but 59 patients (28.10%) experienced postoperative complications and 9 patients (4.28%) died within 30 days of operation. After exclusion of postoperative deaths, the 1-year, 3-year, and 5-year survival rates were 66.21%,36.13% and 24.26%, respectively. Only 1 out of the 12 patients with small cell carcinoma survived more than 3 years. CONCLUSION: Mortality and morbidity were high in pneumonectomy.If we carefully and properly select the patients, enhance postoperative monitoring and perioperative treatment, and combine with chemo-and/or radiotherapy,pneumonectomy will surely be effective for patients with lung cancer.

[Fifty-three cases of valve repair for mitral insufficiency].

OBJECTIVE: To summarize the experience of mitral vavuloplasty for mitral insufficiency in 53 patients. METHODS: Between January 2001 and September 2006,53 patients (31 males and 22 females) with mitral insufficiency underwent mitral valve repair at our hospital. The mean age was (23.8+/-10.4) years, and the mean weight was (43+/-12) kg,including 29 cases of mitral prolapse,17 congenital mitral insufficiency, 5 rheumatic mitral insufficiency,and 2 other etiology. All operations were performed under cardiopulmonary bypass,moderate hypothermia and under the surveillance of transesophagus echocardiography. RESULTS: There were 2 early postoperative deaths,and another patient failed to repair and received valve replacement afterward. Postoperative echocardiography indicated that mitral insufficiency was completely corrected in 42 patients, and residual mitral regurginitation in the other 8 patients. During the follow-up with mean period 13.8 months,41 patients revealed 28 in NYHA classI,13 in Class II. CONCLUSION: Mitral valve repair should be the preferred modes of surgical correction for regurgitate mitral valves. With proper vavuloplasty technique, patients with selective mitral insufficiency patients may achieve satisfactory effect.

[Mechanical valve replacement in children with heart valve diseases].

OBJECTIVE: To determine the surgical point and technique of artificial mechanical valve replacement in children with heart valve diseases. METHODS: From Jan. 1989 to Oct. 2005, 63 children under 15 years received mechanical cardiac valve replacement with cardiopulmonary bypass (CPB). RESULTS: The valve replacement included aortic valve replacement in 20 children, mitral valve replacement in 37 children and combined aortic valve and mitral valve replacement in 6 children. CONCLUSION: The operation mortality was 7.94%(5/63). The follow-up periods were from 4 months to 204 months. The late mortality was 10.34%(6/58). All the other children were in NYHA class I - II. The operation mortality of children with heart valve replacement is higher than that of adults, but it was very effective.

Proteomic analysis of secreted proteins of non-small cell lung cancer.

BACKGROUND & OBJECTIVE: Secreted proteins from cancer cells may be potential serologic biomarkers of cancer. It's important to globally identify secreted proteins of cancer cells. This study was to identify secreted proteins of lung cancer cells. METHODS: Proteins in the conditioned medium of non-small cell lung cancer (NSCLC) cell line A549 was collected and the proteome analysis was subsequently performed. Specific protein spots in A549 cells were identified by peptide mass fingerprints using mass spectrometry and through searching database. The expression of identified secreted proteins was detected by reverse transcription-polymerase chain reaction (RT-PCR) in 15 specimens of NSCLC tissue and paired distant lung tissue. Manganese superoxide dismutase (Mn-SOD) activity in serum and conditioned medium was detected by spectrophotometry. RESULTS: Fourteen secreted proteins were identified, which included peptidyl-prolyl cis-trans isomerase A (PPIA), Mn-SOD, peroxiredoxin 1 (PDX1), phosphatidylethanolamine binding protein (PEBP), glutathione S-transferase P (GSTP1-1), glucose-dependent insulinotropic protein receptor (GIPR), ubiquitin carboxyl-terminal hydrolase isozyme L1 (PGP9.5), alpha enolase (ENO1), dihydrodiol dehydrogenase (DDH), phosphoglycerate mutase 1 (PGAM1), galectin-1 (GAL1). PPIA, DDH, PGAM1, PDX1, PGP9.5, ENO1, and PEBP were overexpressed in cancer tissues. Higher level of Mn-SOD activity was detected in conditioned medium than in control. Serum Mn-SOD activity was significantly higher in NSCLC patients than in healthy controls (P<0.01). CONCLUSIONS: Multiple secreted proteins of A549 cells were identified in this study and the overexpression of ENO1 and PEBP in NSCLC was revealed for the first time. Mn-SOD is secreted serologic marker of NSCLC. The results presented here would provide clues to identify new serologic biomarkers of NSCLC.

[Effect of cortisol on ischemic preconditioning in rabbit hearts].

OBJECTIVE: To determine the effects of reacting cortisol on ischemic preconditioning (IPC). METHODS: Thirty rabbits were randomized into ischemic preconditioning (IPC) group, etomidate (Etom) group, ischemic/reperfusion (IR) group, methylprednisolone (MP) group and a control group. The ratios of infarction size versus risk area (Infarct/Risk) were calculated. The elevations of serum CK activity and cTnI concentrations as well as serum cortisol concentrations were measured. RESULTS: The ratios of Infarct/Risk were 5.9%+/-2.8% for IPC, 11.3%+/-3.6% for Etom, 26.8%+/-4.5% for IR and 18.2%+/-3.7% for MP. The elevations of serum CK activity (U/L) were 255+/-89 for IPC, 314+/-160 for Etom, 855+/-371 for IR and 768+/-404 for MP, the elevations of serum cTnI concentrations (microg/L) were 3.6+/-0.6 for IPC, 5.9+/-2.0 for Etom, 8.1+/-3.6 for IR and 6.1+/-2.3 for MP. Those indicators among groups are all significantly different (P<0.01). Cortisol reacting was markedly diminished in Etom group. CONCLUSION: A blunted cortisol reaction can markedly undermine the benefit of ischemic preconditioning, methylprednisolone show a certain myocardial protection. Cortisol may play an important role in the IPC process.

Proteomics-based identification of secreted protein dihydrodiol dehydrogenase as a novel serum markers of non-small cell lung cancer.

Identification of secreted proteins of lung cancer could provide new candidates of serum biomarkers for cancer diagnosis and prognosis evaluation. In this study, non-small cell lung cancer (NSCLC) cell line A549 was cultured. Proteins in the conditioned medium of A549 were recovered and the proteome analysis was subsequently performed. Secreted proteins of A549 were identified using mass spectrometry and database search. Fourteen human proteins were identified, including peptidyl-prolyl cis-trans isomerase A, manganese superoxide dismutase, peroxiredoxin 1, phosphatidylethanolamine-binding protein, glutathione S-transferase P, PGP9.5, alpha enolase, phosphoglycerate mutase 1, galectin-1 and dihydrodiol dehydrogenase (DDH). DDH was selected for further analysis using RT-PCR, immunoblotting, immunohistochemical staining and ELISA in NSCLC patients. Compared with normal lung tissues, higher DDH mRNA and protein expression level were found in 15 NSCLC cancer tissues (p<0.05). DDH overexpression was identified to be located in cytoplasm and cell membrane by immunohistochemical staining in NSCLC tissue. The serum level of DDH was significantly higher in NSCLC patients (n=64) than nonmalignant lung tumor (n=20) and healthy controls (n=20) (p<0.05). The results show that DDH was one of the secreted proteins in NSCLC. It can serve as a tissue marker and a novel serological marker of NSCLC. Identification of secreted proteins could be a feasible and effective strategy to search potential serum biomarkers of cancer.

[Protective effect of lidocaine on injury alveolar Type II cells induced by LPS in adult rats].

OBJECTIVE: To investigate the effect of lidocaine on LPS induced apoptosis of cultured adult rat alveolar Type II (AT-II) cells. METHODS: Cultured cells were exposed to LPS and lidocaine for 24 hours. Apoptosis and necrosis rates of cells were detected by flow cytometry and electron microscope. The activity of lactic dehydrogenase (LDH) was analyzed by using LDH kits. RESULTS: LPS induced the AT-II cell injuries by increasing not only the necrosis and apoptosis rates but also the LDH release of cultured AT-II in vitro. Lidocaine decreased the necrosis and apoptosis rates of AT-II cells. CONCLUSION: Lidocaine can directly inhibit the apoptosis and necrosis induced by LPS in cultured AT-II cells.

[Direct observation of the effect of mitral valvuloplasty on beating heart].

OBJECTIVE: To summarize the method and experience of direct observation of the effect of mitral valvuloplasty on beating heart. METHODS: Twelve patients with mitral regurgitation underwent mitral valvuloplasty. When the heart was beating slowly before releasing the aorta, the morphology and function of the mitral valve were evaluated by direct observation. If the results were unsatisfactory, further techniques were used to reach the goal. RESULTS: During the operation 2 patients needed further techniques to make the mitral valve work well. No operative death and only 1 complication (low cardiac output syndrome) occurred. After 1 month follow-up later, mild regurgitation was found in 5 patients, moderate regurgitation in 1 and no obvious regurgitation in 6. CONCLUSION: Evaluating the effect of mitral valvuloplasty by direct observation on beating heart is practicable.

[Surgical treatment of cor triatriatum in 15 patients].

OBJECTIVE: To review the clinical data of pathological morphology, diagnosis, surgical treatment of cor triatriatum in 15 patients. METHODS: Fifteen patients with a mean age of (14.6+/-10.3) years (range from 6 months to 40 years) were performed operations under extracorporeal circulation. Fourteen of the patients had cor triatriatum sinister, and 1 had cor triatriatum dexter; 12 of the 15 patients had other cardiac abnormalities. The excision of the fibromuscular membrane was accomplished through a right atrial incision in all of the 14 cases, and the associated abnormalities were corrected at the same time. RESULTS: One patient died after the operation, and the other survivors had good outcome. CONCLUSION: Operation is necessary if the diagnosis is clear. The patients generally have good prognosis. Surgical results of cor triatriatum depend on the complexity of associated defects and the adequacy of the repair.

[Protective mechanism of ischemic preconditioning to the lung ischemia-reperfusion injury].

OBJECTIVE: To determine the protective mechanism of preconditioning to the lung injury induced by ischemia-reperfusion. METHODS: Twelve pigs were randomly divided into 2 groups: control group ( Group C) and ischemic preconditioning group ( Group IP). The concentration of superoxide distrautase (SOD) and malondialdehyde (MDA) in circuit were checked before and after the perfusion to reflect the lipid peroxidation in the lungs. Left lung biopsies were performed immediately after the perfusion and 1 hour postperfusion for histologic examination. The ICAM-1 expression was assessed by immunohistochemical analysis with Envision method and the mRNA expression of ICAM-1 was analyzed by RT-PCR. RESULTS: SOD in Group IP was much higher than that in Group C (P < 0.01 ). MDA in Group IP was much lower than that in Group C ( P < 0.01 ). The lung histologic examination showed that Group C was significantly more serious than Group IP in pulmonary edema, inflammatory cell infiltration, mild focal hemorrhage, and alveolar disruption. The expression of ICAM-1 of lung tissue obviously decreased in Group IP than that in Group C (P <0.01 ). The expression of ICAM-1 mRNA of lung tissue was significantly lower in Group IP than that in Group C (P < 0.01 ). CONCLUSION: Lung ischemic preconditioning can reduce the lung injury. The mechanisms of the protective effects of the IP may be related to the increase of SOD and the decrease of MDA. The preconditioning down-regulated the ICAM-1 expression is one of the mechanisms in reducing the lung injury.

[Protective effects of hypoxic preconditioning on the hypoxic/reoxygenation injury of neonatal rats cardiomyocytes].

OBJECTIVE: To investigate the protective effects of hypoxic preconditioning (HPC) on myocardial cells, and to determine the protective mechanism. METHODS: In the HPC model, the cells viability, productions of malondialdehyde (MDA), contents of ATP, lactate dehydrogenase (LDH), creatine kinase (CK), and expressions of intercellular adehesion molecule-1 (ICAM-1) were measured in the cultured myocardial cells of rats. RESULTS: HPC increased cells viability, attenuated the formation of lipid peroxides and ATP depletion, decreased the leakage of LDH and CK from the cells, and inhibited the increase of expression in intercellular adhesion molecule-1. The blocking agent of ATP sensitive K+ channel (K(ATP) channel), glibenclamide (Glb) completely abolished these protective effects of HPC. CONCLUSION: The K(ATP) channel is an important effector of the protective effects of HPC, and the mechanism of the protective effects of HPC might be related to the inhibition of ICAM-1 expression.

[Preservation of the entire mitral subvalvular apparatus during mitral valve replacement in patients with mitral stenosis].

OBJECTIVE: To evaluate the surgical technique and the effect of preservation of the entire mitral subvalvular apparatus during mitral valve replacement (MVR) in patients with mitral stenosis. METHODS: MVR was performed on 56 patients with mitral stenosis. Of them, 11 had complete preservation of the entire mitral subvalvular apparatus (group A) with the technique of preservation of 'button-shaped' transplantation, 25 had the preservation of the posterior leaflet only (group B), and the other 20 underwent the conventional MVR (group C). Pre- and postoperative hemodynamic parameters were measured to determine the left ventricular performance. Echocardiography was performed preoperatively, at the time of discharge, and 3 to approximately 6 months postoperatively to determine the dimensions and ejection function. RESULTS: Cardiac index, stroke volume index and left ventricular stroke work index postoperatively in group A were better than those in group B and C (P < 0.05). Echocardiographic measurements postoperatively showed the increase of LVESD, LVEDD in group B and C, but a better LVL and LVFS in group A (P < 0.05), and LVEF was more pronounced in group A (P < 0.05). There appeared a long cross-clamp time in group A, but the total CPB time was not significant among the three groups. CONCLUSION: The surgical skills of preservation with 'button-shaped' transplantation may be a practical choice for patients with mitral stenosis. The preservation of entire mitral subvalvular apparatus improves the left ventricular functions after MVR in patients with mitral valve stenosis.