Effects of exercises with different durations and intensities on mitochondrial autophagy and FUNDC1 expression in rat skeletal muscles / 生理学报

The aim of the present study was to investigate the effects of exercises with different durations and intensities on mitochondrial autophagy and FUNDC1 in rat skeletal muscles. Sixty male Sprague-Dawley rats were randomly divided into 2- and 4-week control groups (Con), moderate-intensity exercise groups (M-ex groups, treadmill exercise, 16 m/min, 1 h/d, 6 d/week), and high-intensity exercise groups (Hi-ex groups, treadmill exercise, 35 m/min, 20 min/d, 6 d/week). The bilateral soleus muscles were separated after the intervention, and paraffin sections were prepared for transmission electron microscopy. ELISA method was used to detect the content of citrate synthase (CS). The co-localizations of microtubule-associated protein 1 light chain 3 (LC3)/cytochrome c oxidase IV (COX-IV), FUNDC1/COX-IV and LC3/FUNDC1 were observed by immunofluorescent staining in frozen sections. The skeletal muscle mitochondria were extracted, and the expression of autophagy-related proteins, including AMPKα, p-AMPKα, Unc-51 like kinase 1 (ULK1), FUNDC1, LC3 and p62, were detected by Western blot. The results showed that exercise increased mitochondrial function, i.e. peroxisome proliferator-activated receptor γ co-activator-1α (PGC-1α), COX-I protein expression levels and CS content. There was no difference of mitochondrial function parameters between 2-week M-ex and 2-week Hi-ex groups, while mitochondrial function of 4-weeks Hi-ex group was significantly lower than that of 4-week M-ex group. Under the same exercise intensity, mitochondrial autophagy activation in skeletal muscle of 4-week exercise was higher than that in 2-week exercise group; Under the same duration of exercise, mitochondrial autophagy activation of Hi-ex group was higher than that in M-ex group. Both 2- and 4-week exercise intervention increased LC3/COX-IV, COX-IV/FUNDC1, and FUNDC1/LC3 co-localizations. Exercise increased LC3-II/LC3-I ratio, down-regulated p62 protein expression level, up-regulated FUNDC1, ULK1 protein expression levels and AMPKα phosphorylation, and the changes of these proteins in 4-week Hi-ex group were significantly greater than those in 4-week M-ex group. These results suggest exercise induces mitochondrial autophagy in skeletal muscles, and the activity of autophagy is related to the duration and intensity of exercise. The induction mechanism of exercise may involve the mediation of FUNDC1 expression through AMPK-ULK1 pathway.

Effects of melatonin on oxidative stress and apoptosis-related gene signaling pathways following testicular torsion in rats / 中华男科学杂志

Objective@#To investigate the effects of melatonin on the oxidative stress and signaling pathways of apoptosis-related genes following testicular torsion/detorsion in male rats.@*METHODS@#Twenty-four healthy male Sprague-Dawley rats were randomly divided into a control, a torsion and a melatonin group of equal number. The torsion model was made in the animals of the latter two groups by 720° torsion of the left testis for 2 hours. The rats of the torsion and melatonin groups received intraperitoneal injection of isotonic saline and melatonin (17 mg/kg) respectively at 15 minutes prior to detorsion. At 24 hours after modeling, testis tissues were collected from the rats for detection of the apoptosis of the germ cells by flow cytometry (FCM), analysis of the expressions of Fas, Fas ligand (FasL) and Bax mRNA by quantitative real-time PCR (qRT-PCR), measurement of the cytochrome C content released from the mitochondrion by Western blot, and determination of the total antioxidant capacity (T-AOC) and the levels of myeloperoxidase (MPO) and malodialdehyde (MDA) by spectrophotometry.@*RESULTS@#Compared with the torsion group, the rats treated with melatonin showed significantly increased normal testicular cells (［77.81 ± 6.52］% vs ［88.61 ± 7.93］%, P < 0.05), decreased early apoptotic germ cells (［16.74 ± 3.16］% vs ［6.97 ± 1.65］%, P < 0.05), down-regulated expressions of Fas (［4.52 ± 0.29］ vs ［2.66 ± 0.37］, P < 0.01), FasL (［2.82 ± 0.30］ vs ［1.73 ± 0.18］, P < 0.01) and Bax mRNA (［2.39 ± 0.18］ vs ［1.50 ± 0.14］, P < 0.01), reduced levels of cytochrome C (［1.40 ± 0.38］ vs ［0.67 ± 0.30］, P < 0.01), MPO (［0.52 ± 0.15］ vs ［0.19 ± 0.10］ U/g prot, P < 0.01) and MDA ［6.37 ± 1.73］ vs ［3.98 ± 0.90］ nmol/mg prot, P < 0.01) and elevated T-AOC (［0.76 ± 0.25］ vs ［1.55 ± 0.32］ U/mg prot, P < 0.01).@*CONCLUSIONS@#Melatonin has a significant protective effect on spermatogenesis after testicular torsion by regulating the expressions of apoptosis-related genes and increasing T-AOC in the testis tissue.

An in vitro study of enhancing tendon-bone healing by human amniotic mesenchymal stem cells co-modified with hypoxia-inducible factor 1 alpha and Scleraxis gene / 中国组织工程研究

BACKGROUND: Tendon-to-bone healing is a complex and slow process, which often hinders the therapeutic outcomes. To enhance the tendon-to-bone healing, increasing cell bioactivity on the contact surface is a new strategy. Hypoxia-inducible factor 1α (HIF-1α) can induce the formation of blood vessel and bone tissue via many ways.However,it is unclear whether HIF-1α can strengthen differentiation of human amniotic mesenchymal stem cells (hAMSCs) induced by Scleraxis, a specific marker of tendon, and can be used in tendon and bone repair. OBJECTIVE: To investigate the ability of HIF-1α in enhancing Scleraxis to modify hAMSCs aiming to promote tendon-to-bone healing and its molecular mechanism. METHODS: Passage 3 hAMSCs were infected with AdHIF-1α, AdScx, AdGFP and AdHIF-1α+AdScx. The expressions of related genes of tendon, cartilage and bone tissue were detected at 3 and 7 days after infection. RESULTS AND CONCLUSION: Under the inverted phase contrast microscopy, the passage 3 hAMSCs were in spindle shape and presented with vortex-like adherent growth. Under the transmission electron microscopy, hAMSCs were in oval shape and had clear structure, with abundant endoplasmic reticulum and mitochondria. Fluorescence photographs were taken at 24 hours after adenovirus infection, showing about 50% red fluorescence expression in the AdHIF-1α group, while about 70% green fluorescence expression in the AdScx and AdGFP groups. Real time-PCR results showed that at 3 and 7 days after infection, the mRNA expressions of collagen type I, Fibronectin, RUNX2, VEGF and ALP in the AdHIF-1α+AdScx group, AdHIF-1α group and AdScx group were higher than those in the AdGFP group (P < 0.05); the mRNA expressions of collagen type I, Fibronectin, RUNX2, VEGF and ALP in the AdHIF-1α+AdScx group were significantly higher than those in the AdScx group (P < 0.05). Fluorescence immunohistochemistry results showed that the expression of collagen type I in the AdHIF-1α+AdScx group at 7 days after infection was higher than that at 3 days after infection. To conclude, HIF-1α can enhance the ability of Scleraxis to modify hAMSCs to promote tendon-to-bone healing by upregulating the expressions of molecular markers of tenocytes, chondrocytes and osteocytes.

Molecular pathways of germ cell apoptosis following testicular torsion in rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the molecular mechanism of germ cell apoptosis following testicular torsion in rats.</p><p><b>METHODS</b>Healthy male Sprague-Dawley rats (n = 16) were equally randomized into a control and a torsion group and the models of testicular torsion (720 degrees 2 h) were established. Twenty-four hours later, the apoptosis and count of germ cells were determined by flow cytometry, the expressions of Bax, Fas and Fas ligand (FasL) mRNA semiquantitatively analyzed by RT-PCR and the cytochrome C release detected by Western blot.</p><p><b>RESULTS</b>Compared with the control group, there was an obvious increase in the number of apoptotic germ cells, a marked decrease in that of haploid and tetraploid cells and significantly up-regulated expressions of Bax and Fas/FasL mRNA in the torsion group (P <0.01). The Western blot analysis showed that the cytochrome C release was remarkably increased 24 hours after the detorsion. There were significant differences between the two groups (P <0.01).</p><p><b>CONCLUSION</b>There are two major signaling pathways of cell apoptosis following testicular torsion, intercellular and intracellular. Up-regulated expressions of the apoptosis-related molecules Bax and Fas/FasL and increased cytochrome C release may play an important role in germ cell apoptosis following testicular torsion in rats.</p>

Protective effect of pentoxifylline on spermatogenesis following testicular torsion/detorsion in rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the protective effect of pentoxifylline on spermatogenesis following testicular torsion/detorsion in rats.</p><p><b>METHODS</b>Healthy male Sprague-Dawley rats (n = 24) were divided into three groups randomly, each comprising 8 rats. In Group I, rats underwent a sham operation. In Group II and III, animals were submitted to unilateral 720 degrees testicular torsion, then detorsion in two hours. Infusion of isotonic saline and pentoxifylline into tail vein was initiated 15 minutes prior to relief of torsion in Group II and III respectively. Twenty four hours later, testes were examined for evidence of germ cell apoptosis by the flow cytometry and the level of total antioxidant capability (T-AOC) and malondialdehyde (MDA) by spectrophotometry.</p><p><b>RESULTS</b>Compared with that of group II, the number of apoptotic germ cell and the level of MDA decreased remarkably in Group III, but T-AOC increased significantly (P <0.01).</p><p><b>CONCLUSION</b>Pentoxifylline provided significant rescue of testicular function after acute experimental torsion.</p>

Target-controlled infusion of remifentanil and propofol during operation with suspension laryngoscopy / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To evaluate target-controlled infusion (TCI) of remifentail-propofol and the balanced anesthesia of fentanyl-isoflurane during the operation with suspension laryngscope.</p><p><b>METHODS</b>Sixty ASA I-II patients scheduled for the surgery through suspension laryngoscopy were randomly divided into two groups: TCI group and control group. In TCI group, anesthesia was maintained with TCI remifentanil-propofol which was stopped at the end of operation. The target plasma concentration of remifentanil was set at 6 microg/L and propofol at 3 mg/L. In control group, anesthesia was induced with intravenous fentanl 2.5 microg/kg and propofol 1-2 mg/kg, maintained with fentanl 0.03 microg.kg(-1). min(-1) and 1% isoflurane which was stopped at the end of surgery. Intubation was facilitated with succinylcholine 1-1.5 mg/kg.MAP, HR, ECG, S(p)O(2) and P(ET)CO(2) were monitored during anesthesia. The following parameters were recorded and compared between two groups: (1) the changes in blood pressure (BP), heart rate(HR) and S(p)O(2) at different time point; (2) recovery profile including the time of response to verbal commands, autonomous breathing, tracheal extubation, orientation recovery, discharging from PACU after operation; (3) OAAS scores after operation; (4) postoperative complications; (5) unexpected events and awareness during operation.</p><p><b>RESULT</b>(1) The hemodynamics were stable while the target plasma concentration of remifentanil was set at 6 microg/L and propofol at 3 mg/L. (2) During tracheal intubation, suspension laryngoscope was inserted, and extubation MAP was significantly lower in TCI group than that in control group; (3) There were no significant differences in hemodynamic values and S(p)O(2) of different time points between two groups. Study group was faster than control group on recovery profile including the time of response to verbal commands, autonomous breathing, tracheal extubation, orientation recovery and discharging from PACU. There was respectively one unexpected event in both groups.</p><p><b>CONCLUSION</b>Remifentanil supplemented with isoflurane anesthesia can achieve the optimal hemodynamic stability during the operation with suspension laryngoscopy and better recovery profile from anesthesia than fentanyl.</p>

Combination of lamivudine with thymosin alpha1 in treatment of duck hepatitis B / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To explore the inhibitory effect of combination of lamivudine with thymosin alpha1 (Talpha1) on the replication of duck hepatitis B virus (DHBV).</p><p><b>METHODS</b>Peking ducks of 1 d old were challenged with DHBV-positive serum and used as a duck hepatitis B model. After treated with lamivudine for three months, the ducks were randomly grouped and treated with or without Talpha1 for 8 d. Serum DHBV titrate was observed by semi-quantitative PCR, and inflammation and degeneration of hepatocytes were observed by pathology examination.</p><p><b>RESULTS</b>The serum DHBV titrate was significantly reduced (4483.2+/-5193.4 compared with 9351.8+/-5059.6) after lamivudine treatment, and it was reduced more significantly(1692.2+/-589.2) after combination treatment with Talpha1. Lamivudine reduced the degeneration degree of hepatocytes (3.2+/-0.8 compared with 4.6+/-0.5) and the inflammation degree of liver (6.2+/-3.3 compared with 8.6+/-2.8). The combination treatment with Talpha1 increased liver inflammation degree (9.0+/-5.2).</p><p><b>CONCLUSION</b>Both Talpha1 and lamivudine may reduce the replication of DHBV in Peking ducks and combination treatment may have the better anti-virus effect and enhance immune response in liver.</p>

Immunologic changes to autologous transfusion after operational trauma in malignant tumor patients: neopterin and interleukin-2 / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>To estimate the impact of autologous transfusion on the status of perioperative immune activation in malignant tumor patients. The Serum Neopterin and Interleukin-2 (IL-2) were measured.</p><p><b>METHODS</b>Sixty patients undergoing elective radical resection for malignant stomach tumor were enrolled in the prospective study and assigned to the following groups: (1) Group A received autologous transfusion. (2) Group H received allogeneic transfusion. The perioperative course (Before induction of anesthesia, after operation and 5 d after operation) of Neopterin and IL-2 was compared.</p><p><b>RESULTS</b>In group A, Serum Neopterin was significantly lower than baseline after operation and IL-2 had no significant changes. In group H, both Serum Neopterin and IL-2 were significantly lower than baseline after operation and 5 d after operation. Compared with group A, Serum Neopterin was significantly lower than baseline after operation and 5 d after operation and IL-2 was significantly lower than baseline 5 d after operation.</p><p><b>CONCLUSION</b>Autologous transfusion decreased the perioperative immune suppression in malignant stomach tumor patients.</p>

Effects of ulinastatin on renal ultrastructure after ischemia-reperfusion in rats / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of ulinastatin on renal function and ultrastructure changes after renal ischemia-reperfusion in rats.</p><p><b>METHODS</b>Acute ischemic renal injury model was established (45 min of bilateral renal ischemia and reperfusion for 24 h). Thirty Male SD rats were randomly divided into 3 groups: sham operation group (control group or group C, without renal ischemia), renal ischemia-reperfusion group (ischemia-reperfusion group or group I, without ulinastatin), renal ischemia-reperfusion and ulinastatin intravenous injection group (ulinastatin group or group U). BUN level, serum creatinine values and renal ultrastructure were measured.</p><p><b>RESULTS</b>Serum creatinine (167 +/- 39) micromol/L and BUN concentration (21 +/- 7) mmol/L in group I were significantly higher than those in group U: serum creatinine (116 +/- 13) micromol/L and BUN concentration (14.1 +/- 2.6) mmol/L (P < 0.05). The renal ultrastructure was greatly injured in group I, meanwhile, it was obviously ameliorated in group U.</p><p><b>CONCLUSION</b>Ulinastatin greatly improved renal function and provides remarkable protection on renal ultrastructure after ischemia-reperfusion of kidney in rats.</p>

Reversal of multidrug resistance gene MDR1 and MRP of drug-resistant human hepatocellular carcinoma cells SMMC-7721/ADM with antisense phosphorothioate oligonucleotides / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the reversal effect of gene MDR1 and MRP with combinational antisense phosphorothioate oligonucleotide on Drug-resistant human hepatocellular carcinoma cells SMMC-7721/ADM.</p><p><b>METHODS</b>SMMC-7721/ADM was transfected with synthetic antisense phosphorothioate oligonucleotides complementary to gene MDR1 and MRP mediated by Lipofectamine. Drug sensitivity was measured by MTT assay, Fluorescence intensity of cells was determined by flow cytometric analysis, RH123 and DNR retention was assayed by confocal scanning laser microscopy.</p><p><b>RESULTS</b>ASODN of MDR1+MRP increased the sensitivity of SMMC-7721/ADM to chemotherapeutic drug more significantly than that any of MDR1 and MRP did separately. But they did not enhance the inhibition expression of protein of p190 or p170.</p><p><b>CONCLUSION</b>Drug-resistance could be reversed significantly when antisense phosphorothioate oligonucleotide of MDR1+MRP were transfected into drug-resistant human hepatocellular carcinoma cells SMMC-7721/ADM together.</p>

Germ cell apoptosis and expression of Bcl-2 and Bax following testicular torsion/detorsion in rats / 中华男科学杂志

<p><b>OBJECTIVES</b>To investigate the relationship between germ cell apoptosis and expression of Bcl-2 and Bax in experimental torsed/detorsed testes of the adult male rats.</p><p><b>METHODS</b>Thirty healthy male Sprague-Dawley rats were divided into torsion group (n = 15) and control group (n = 15) randomly. Animals were submitted to unilateral 720 testicular torsion, then detorsion were done in two hours. Three days later, the evidence of germ cell apoptosis was detected by the TdT-mediated dUTP-biotin nick end labeling (TUNEL) technique. The expression of Bcl-2 and Bax were detected by immunohistochemical method.</p><p><b>RESULTS</b>In the torsed testes, the apoptosis index of germ cell and Bax expression significantly increased compared with that in the control group (P < 0.01) while Bcl-2 expression obviously decreased (P < 0.01). The apoptotic cells were mostly pechytene spermatocytes and round spermatides.</p><p><b>CONCLUSIONS</b>The germ cell apoptosis is highly associated with expression of Bcl-2 and Bax in experimental testicular torsion. Bcl-2/Bax plays an important role in germ cell apoptosis.</p>