Heart rate variability(HRV)analysis and its circadian rhythm(CR)were determined in 58patients with type 2 diabetes mellitus with lower extremity neuropathy(diabetic neuropathy group), 59 patients with type 2 diabetes mellitus without lower extremity neuropathy(diabetes group), and 50 healthy controls according to 24-hour Holter recording. Frequency domain parameters of HRV were significantly decreased in both diabetes groups. Frequency domain parameter of HRV in healthy controls,and daytime/nighttime difference were statistically significant. CR of HRV was changed in diabetes group and disappeared in diabetic neuropathy group. Impaired and seriously impaired autonomic nervous function developed in type 2 diabetes mellitus without and with lower extremity neuropathy respectively.

BACKGROUND: Umbilical cord blood mesenchymal stem cells (UCB-MSCs) can differentiate into various types of cells under certain condisions, and easily proliferate in vitro. However, UCB-MSCs have long establishment time and low frequency.OBJECTIVE: To in vitro isolate and culture UCB-MSCs, and induce its differentiation into osteoblasts.DESIGN, TIM E AND SETTING: The in vitro cytological study was performed at the Laboratory of the Medical College of Qingdao University from June 2008 to January 2009.MATERIALS: UCB was obtained from term normal delivery women at the Department of Gynaecology and Obstetrics, Qingdao Municipal Hospital.METHODS: Human UCB-MSCs were isolated and cultured in vitro by Percoll density gradient. When reached 90% confluency,UCB-MSCs were digested by trypsin for subculture. At the third passage, UCB-MSCs at 1×106 were incubated. When reached 50% 60% cenfluency, UCB-MSCs were treated with DMEM supplemented with 0.1 μmol/L dexamathasone, 10 mmol/Lβ-sodium glycerophosphate and 50 μmol/L vitamin C. UCB-MSCs in the control group were incubated in low glucose DMEM.MAIN OUTCOME MEASURES: Growth and proliferation of MSCs were observed under the inverted microscope. Cell surface marker expression and cell growth curve were measured by flow cytometry. Cell ultrastructure was observed under the transmission electron microscope. Differentiation of UCB-MSCs into osteoblasts was determined by Won Kossa staining and alkaline phosphatase staining.RESULTS: Primary cultured UCB-MSCs had similar morphology to bone marrow mesenchymal stem cells. After passage, cell morphology was even, presenting spindle shape. UCB-MSCs at passage 3 highly expressed CD29, CD44, CD13, but did not express CD34. Growth latency was 2-3 days. Cells entered logarithm proliferation phase at days 3-4, and platform phase 1 month later. Nuclei presented round or irregular, with clear nuclear membrane, 1-2 nucleoli, rough chromatin, abundant organelles and microvilli. UCB-MSCs at passage 3 were gradually confluent following 3 days of osteogenic induction, with the presence of pavement-stone shape. 14 days later, calcified nodules by Von Kossa staining, and cells were positive for alkaline phosphatase staining. In the control group, no calcified nodules were found, and cells were negative for alkaline phosphatase staining.CONCLUSION: UCB-MSCs can be successfully isolated by Percoil density gradient, and induced to differentiate into osteoblasts in vitro.

Objective To investigate the anti - stress activity of thymopentin ethyl ester. Methods Anti - stress activity was evaluated by heat stress model and chronic uncontrolled stress model. Heat stress model;of 60 KM mice were divided into six groups. Except the controlled group, the other mice were maintained in 42℃ for 1h. Chronic uncontrolled stress model:of 60 KM mice were divided into six groups. Except the controlled group, the other mice were given three different stimulations once a day for continuous 21 d. The controlled group and model group were injected saline 0.2ml, and the three test groups were respectively injected thympentin ethyl ester at 2mg/kg,0.2mg/kg, 0.02mg/kg subcutaneously. The positive controlled mice were given thymopentin 0.2mg/kg subcutaneously. At the end of the experiment, plasma corticosteroid, IL -2 and SOD were determined according to the kit instructions. Results The activities of thymopentin ethyl ester in suppressed corticosteroid up - regulation and the elevated plasma IL - 2 and SOD level were more significant than those of thymopentin(P

Objective To investigate the effects of postconditioning with α7 nicotinic acetylcholine receptor (α7nAChR) agonist and ischemia on myocardial ischemia-reperfusion (IR) injury in rats. Methods Fifty adult male SD rats weighing 290-320 g were randomly divided into 5 groups ( n = 10 each): Ⅰ sham operation group, Ⅱ IR group, Ⅲ ischemic postconditioning group, Ⅳ α7nAChR agonist postconditioning group and Ⅴpostconditioning with α7nAChR agonist and ischemia group. Myocardial I/R was induced by ligation of anterior descending branch of left coronary artery for 30 min followed by 1 80 min of reperfusion. In group] the anterior descending branch was only exposed but not ligated. In group Ⅲ the hearts were subjected to 3 episodes of 10 second ischemia at 10 second intervals at the end of 30 min ischemia before 180 min reperfusion, Intraperitoneal PNU282987 2.4 mg/kg was injected at the end of 30 min ischemia before 180 min reperfusion in group Ⅳ and Ⅴ .Blood samples were taken from right internal jugular vein at 180 min of reperfusion. Then the rats were killed and hearts removed to determine the concentrations of serum cardiac troponin-I (cTnI), TNF-α and high-mobility group box 1 (HMGB1) by ELISA. The infarction size was measured by Evans blue and triphenyltetrazolium chloride staining. Results The infarction size was significantly larger in the other groups and concentrations of serum cTrI, TNF-α and HMGB1 were significantly higher in group Ⅱ than in group Ⅰ ( P ＜ 0.05). The infarction size was significantly smaller and concentrations of serum cTnI, TNF-α and HMGBI were significantly lower in group Ⅲ, Ⅴ than in group Ⅱ (P ＜ 0.05). The infarction size was significantly smaller in group Ⅴ and concentrations of serum cTnI, TNF-α and HMGB1 were significantly lower in group Ⅳ and Ⅴ than in group Ⅲ (P ＜0.05). The infarction size was significantly smaller and concentrations of serum cTnI, TNF-α and HMGB1 were significantly lower in group Ⅴ than in group Ⅳ ( P ＜ 0.05 ). Conclusion Postconditioning with α7nAChR agonist and ischemia can reduce myocardial I/R injury and the efficacy is better than that of α7nAChR agonist postconditioning or ischemic postconditioning alone.

Objective To study the process and mechanism underlying Wallerian degeneration of the central nervous system after stroke.Methods A case suspected of stroke with bilateral symmetrical lesions in middle cerebellar peduncles (MCPs) was described.The etiology of bilateral MCP abnormal signals on MR was analyzed according to the clinical process and neuroanatomy.Results Unilateral paramedian pontine infarction,covering the crossing area of pontocerebellar fibers, would cause bilateral secondary degeneration of MCPs,with hyperintense signals on T2-,Flair and diffusion-weighted images.Conclusions Wallerian degeneration of projecting system is a common sequel after stroke and should not be misdiagnosed as other diseases.

Objective To observe the efficacy and safety of endoscope and laparoscope in treatment of acutebiliary pancreatitis.Methods 84 patients with acute biliary pancreatitis from January 2010 to January 2014 in our hospital during treatment were randomly divided into the study group and control group according to the random num-ber table methods,42 cases in each group.The control group was treated with conservative treatment of department of internal medicine,while the study group was treated with duodenoscopy combined with laparoscopy.To observe and compare the efficacy and adverse reactions of two groups.Results Temperature returned to normal time,abdominal pain relief time,recovery time of amylase in urine,recovery time of biochemical index,hospitalization time and cost of hospitalization of the study group after treatment were (3.44 ±1.63)d,(4.73 ±1.12)d,(7.82 ±3.31)d,(9.81 ± 6.42)d,(12.43 ±4.61 )d,which were significantly lower than (5.94 ±2.43)d,(7.53 ±2.12)d,(11.72 ± 2.21)d,(18.41 ±8.62)d,(20.33 ±6.21)d of the control group(t =3.90,4.02,4.26,4.50,5.01,all P <0.05). 2 cases of the study group occured biliary tract bleeding,2 cases with pancreatic pseudocyst,1 case with wound infec-tion,there was no recurrence,the incidence rate of adverse reaction was 11.90%;3 cases of the control group after treatment occurred pancreatic pseudocyst,2 cases with abdominal cysts,5 cases with recurrence,1 case with acute respiratory distress syndrome (ARDS),1 case with renal failure,the incidence rate of adverse reactions was 28.57%, which of the control group was significantly higher than the study group (χ2 =4.21,P <0.05 ).Conclusion Duodenal endoscopy combined with laparoscopy has good clinical effect,shorter hospitalization time,less cost,less adverse reaction and the recurrence rate is low in treatment of acute biliary pancreatitis,which has high application value.

ObjectiveTo investigate the effects of postconditioning with electric vagal stimulation on myocardial ischemia-reperfusion (I/R) injury in rats.MethodsSixty male SD rats weighing 250-350 g were randomly divided into 3 groups (n ＝ 20 each):group sham operation (group S); group myocardial I/R (group I/R) and group electric vagal stimulation postconditioning (group POES).Myocardial I/R was induced by occlusion of left anterior descending branch of coronary artery for 30 min followed by 120 min reperfusion in groups I/R and POES.In group POES right cervical vagus nerve trunk was stimulated for 30 min with continuous electric rectangular pulses (2 ms,10 Hz) starting from 15 min of myocardial ischemia.The voltage of the pulses was adjusted to decrease HR by 10％ of the baseline HR before stimulation.MAP,HR and RPP (MAP× HR) were recorded before (baseline) and at 1 and 10 min of ischemia and 30,60 and 120 min of reperfusion.Arterial blood samples were collected from 10 rats in each group at 120 min of reperfusion for determination of serum concentrations of cTnI,CK-MB,TNF-a,high mobility group box 1 protein (HMGB1),ICAM-1,IL-1,IL-6 and IL-10 (by ELISA).The animals were then sacrificed and myocardial infarct size was measured by Evans blue and TTC staining.Another 10 rats were sacrificed at 120 min of reperfusion for determination of myocardial contents of TNF-α,HMGB1,ICAM-1,IL-1,IL-6 and IL-10 (by ELISA).ResultsI/R induced myocardial infarct,significantly increased serum concentrations of cTnI,CK-MB,TNF-α,HMGB1,ICAM-1,IL-1 and IL-6 and significantly increased myocardial contents of TNF-α,HMGB1,ICAM-1,IL-1,IL-6 and IL-10 in both ischemic and non-ischemic regions in group I/R as compared with group S.Electric vagal stimulation significantly decreased myocardial infarct size and serum concentrations of cTnI,CK-MB,TNF-α,HMGB1,1CAM-1,IL-1 and IL-6 in group POES compared with group I/R.Myocardial contents of TNF-α,HMGB1,ICAM-1,IL-1 and IL-6 were significantly decreased while myocardial IL-10 content was increased in both ischemic and non-isehemic regions in groups POES compared with group I/R.ConclusionPostconditioning with electric vagal stimulation can attenuate myocardial I/R injury by inhibiting inflammatory response in rats.

Objective To explore the operation opportunity of cholecystectomy for senile patients with acute cholecystitis combined with underlying disease.Methods 82 elderly patients with acute cholecystitis complicated with underlying diseases underwent cholecystectomy were selected,they were divided into two groups according to the different operation time:the early operation group and the delayed operation group,41 cases in each group.The patients in early operation group were given operation in 72h,while the delayed operation group was given operation more than 72h after attack.The operation time,intraoperative blood loss,intraoperative tissue adhesions,intraoperative exudation,hospitalization time,cost of hospitalization and complications of the two groups were observed and com-pared.Results The operation time of the two groups were (45.32 ±1 2.23)min,(50.89 ±1 3.34)min,there was no significant difference (P >0.05),the intraoperative tissue adhesions of the delayed operation group was more than the early operation group,and the intraoperative hemorrhage and exudation in the early operation group were significantly higher than those of the delayed operation group (P <0.05),and the hospital stay and cost of hospitalization were sig-nificantly lower than those of the delayed operation group (P <0.05).1 case in the early operation group occurred pulmonary infection,3 cases with incision infection,the incidence of complications was 9.76%;1 case in the delayed operation group underwent laparotomy operation,2 cases in the delayed operation group had pulmonary infection,inci-sion infection in 4 cases,the incidence rate of complication was 1 4.63%,there was no significant difference between the two groups (P >0.05).Conclusion The optimal operation timing of acute cholecystitis combined with basic dis-eases in the elderly patients underwent cholecystectomy is within 72h.For patients who can not be immediately given operation due to various reasons,delayed cholecystectomy is safe and feasible.

Objective To observe the effects of intensive lower extremity motor control training on the motor function of lower extremity, balance and walking capability of stroke patients. Methods 40 stroke patients were randomized into the intervention group (n=20) and control group (n=20). Both groups received the routine rehabilitation, while the intervention group accepted the lower extremity motor control training in addition for 6 weeks. Before and after 6 weeks of treatments, they were assessed with Fugl-Meyer Assessment of lower extremity (FMA-L), Berg Balance Scale (BBS), Holden Functional Ambulation Category (FAC); their gaits were analysesd with footprint. Results Both groups improved significantly in scores of FMA-L, BBS, FAC and average step length, stride width, walking velocity (P<0.01) after treatment, and improved more in the intervention group than in the control group (P<0.05). Conclusion Intensive motor control training of lower extremity may promote the recovery of motor function of lower extremities, balance and walking ability of stroke patients.

OBJECTIVETo investigate the effects of 50 Hz magnetic fields (MF) on DNA double-strand breaks in human lens epithelial cells (hLECs).

METHODSThe cultured human lens epithelial cells were exposed to 0.4 mT 50 Hz MF for 2 h, 6 h, 12 h, 24 h and 48 h. Cells exposed to 4-nitroquinoline-1-oxide, a DNA damage agent, at a final concentration of 0.1 micromol/L for 1 h were used as positive controls.After exposure, cells were fixed with 4 % paraformaldehyde and for H2AX (gamma H2AX) immunofluorescence measurement. gamma H2AX foci were detected at least 200 cells for each sample. Cells were classified as positive when more than three foci per cell were observed. Mean values of foci per cell and percentage of foci positive cells were adopted as indexes of DNA double-strand breaks.

RESULTThe mean value of foci per cell and the percentage of gamma H2AX foci positive cells in 50 Hz MF exposure group for 24 h were (2.93 +/-0.43) and (27.88 +/-2.59)%, respectively, which were significantly higher than those of sham-exposure group [(1.77 +/-0.37) and (19.38+/-2.70)%, P <0.05], and the mean value of foci per cell and the percentage of gamma H2AX foci positive cells in 50 Hz MF exposure group for 48 h were (3.14 +/-0.35) and (31.00 +/-3.44)%, which were significantly higher than those of sham-exposure group (P <0.01). However there was no significant difference between 50 Hz MF exposure groups for 2 h, 6 h, 12 h and sham-exposure group for above two indexes (P >0.05).

CONCLUSION0.4 mT 50 Hz MF exposure for longer duration might induce DNA double-strand breaks in human lens epithelial cells in vitro.

Cells, Cultured ; DNA ; radiation effects ; DNA Breaks, Double-Stranded ; radiation effects ; DNA Damage ; radiation effects ; DNA Repair ; radiation effects ; Electromagnetic Fields ; Epithelial Cells ; metabolism ; radiation effects ; Humans ; Lens, Crystalline ; cytology