AIM:To observe the relation between astroglial activation state and ischemic tolerance in the gerbil hippocampus. METHOD: Bilateral occlusion of common carotid arteries and immunofluorescent methods in the gerbil hippocampal tissue. Slices were used. The morphological changes of the neurons were observed by light microscopy.RESULTS:Pretreatment with 2 minute bilateral carotid artery occlusion produced protective effects of CA 1 neurons. Glial fibrillary acidic protein (GFAP) staining showed weak positive cells in control group. Most of GFAP positive cells showed weak and middle positive cells after recirculation 1d and 2d following ischemic 3.5 min and preconditioning of the brain with sublethal ischemia respectively.CONCLUSION:Astroglial played an important role in cerebral ischemia. It is possible that state of astroglial activation related to neuronal survival in ischemic tolerance.

Objective To observe the effects of electro-acupuncture on the expression and inhibition of DNA binding protein 2 (Id2) and myelin basic protein (MBP),and to explore the mechanism of remyelinization after compressive spinal cord injury (CSCI) in rats.Methods Fifty-four SD rats were randomly divided into a control group and a treatment group,and each was further subdivided into 3 time point subgroups:3,7 and 14 days.There were 9 rats in each subgroup.The CSCI models were made with a self-designed method.The acupuncture points Jiaji (EX-B2),bilateral Zusanli (ST36) and Taixi (KI3) were selected for treatment.Electro-acupuncture (continuous wave,2 Hz,1.5 V)was applied to the bilateral Zusanli (ST36) and Taixi (KI3) points.The control group received the injury but no treatment.The changes in the ultrastucture of the nerve fibers＇ white matter were de-termined by transmission electron microscopy (TEM).The alterations in the expression of MBP and Id2 were observed by double labeled immunofluorescence and Western blotting on the 3rd,7th and 14th day after the injury.Results TEM showed that the myelin sheaths in the control group had degenerated,swollen,and even broken down after CSCI.Changes to the myelin sheaths in the treatment group were milder than those in the control group.The immunofluorescence results showed the amount of Id2-immunoreactive oligodendrocytes in the control group to be (20 ±2) on the 3rd day after CSCI,becoming (16 ± 1) on the 14th day.The differences among the 3 control subgroups were not statistically significant.The amount of Id2-immunoreactive oligodendrocytes in the treatment group was (13 ± 1) on the 3rd day,reaching a minimum the 14th day.The differences among the 3 treatment groups were statistically significant.The differences compared with the control group at the same time points were also statistically significant.Western blotting showed that the expression of Id2 in the contrast and treatment groups was (1.12 ±0.12) and (0.67 ±0.01) respectively on the 3rd day after CSCI,and both decreased with time.The expression of Id2 in both groups reached their minima ((0.86 ±0.02) and (0.25 ±0.01) respectively) on the 14th day.The difference between the treatment groups and the contrast group was statistically significant at each time point.The expression of MBP in the contrast and treatment groups at day 3 was (0.44 ± 0.02) and (0.67 ± 0.04) respectively,and these increased with time.The expression of MBP in both groups peaked at the 14th day (at (0.95 ± 0.04) and (1.74 ± 0.09) respectively).These differences were again statistically significant.Conclusion Electro-acupuncture can regulate the expression of Id2 and MBP after CSCI.The down-regulation of Id2 which controls MBP negatively and the up-regulation of MBP may contribute to remyelination in the injured spinal cord.

Objective To investigate the expression of microRNA-34a (miR-34a) and silent information regulator 1 (SIRT1) in human lens epithelial cells under H2O2-induced oxidative stress.Methods Different concentrations of H2O2 (0 μmol · L-1,100 μ mol· L-1,200 μmol · L-1,300 μmol · L-1,and 400 μmol · L-1) were used to stimulate SRA01/04 cells for 24 hours.Cell viability was measured using cell counting kit-8 (CCK-8) assay.Cell apoptosis was detected by flow cytometry.Expression levels of miR-34a/SIRT1 were measured by RT-PCR.Results CCK-8 assay showed that a certain concentration range of H2O2 had a proliferation inhibition on SRA01/04 cells.There was a dose response relationship between 100 μmol · L-1 and 400 μmol · L-1.Compared with 0 μmol · L-1 H2O2 group,the difference was statistically significant (all P ＜ 0.01).According to flow cytometry results,apoptotic rate of SRA01/04 cells in control group and H2O2(100-300 μmol · L-1) groups were (6.1 ± 1.2)％,(26.3 ± 1.8)％,(32.5 ± 2.2) ％,and (64.7 ± 5.3) ％.Compared with 0 μmol · L-1 H2 O2 group,the differences were statistically significant (all P ＜ 0.01).RT-PCR test results showed that the expression of miR-34a increased significantly in a dose-dependent manner after the SRA01/04 cells treated with different concentrations of H2O2,while SIRT1 expression level was decreased,there were significant differences compared with control group (all P ＜ 0.001).Conclusion There is a significantly increase of miR-34a and decrease of SIRT1 in human lens epithelial cells under the oxidative stress of a certain concentration of H2O2.Down-regulated expression of miR-34a can increase the survival rate of human lens epithelial cells under H2O2-induced oxidative stress.

Objective　To observe the clinical effects of human amniotic membrane as an alternative substrate for wrapping hydroxyapatite ocular prosthesis implants.Methods　After enucleation, hydroxyapatite orbital implants wrapped by human amniotic membrane were implanted with four rectus sutured to the implants .Among them, 4 cases were given primary implantation and 3 cases secondary implantation.Results　After 8 to 39 week follow-up, it was found that in 6 cases, the eyelids were plump and prosthesis moved well, but one implant was removed because of failure of scleral patch graft for implant exposure.Conclusion　The clinical results were satisfactory for ocular prosthesis implantation of hydroxyapatite sphere wrapped by human amniotic membrane, which may be considered as an alternative substrate for ocular prosthesis implants wrapper.

Objective To investigate the role of demyelination and the alteration of chondrotin sulfate proteoglycan (CSPG,NG2) expression after compression injury of the spinal cord (CSCI).Methods Seventy-five adult Sprague-Dawley rats were randomly divided into a normal group,a sham-operation group,a CSCI 1 day group,a CSCI 3 day group,and a CSCI 7 day group.There were 15 rats in each group.The injuries in the CSCI groups were inflicted using a technique devised in our laboratory.Basso-Beattie-Bresnahan (BBB) neurological function assessment was used to assess the rats' motor function,osmic acid staining and transmission electronic microscopy (TEM)were used to observe any pathological changes of myelinated nerve fibers in the white matter at 1,3 and 7 days after CSCI.The amount of myelinated nerve fibers in the posterior funiculus of the spinal cord and the ratio of myelin sheath thickness to axon diameter (the G-ratio) were calculated.Any alteration in NG2 expression was observed by Western blotting.Results The average neurological function assessment scores in the CSCI groups were (1.23 ±0.45),(0.65 ± 0.35) and (0.00 ± 0.00) respectively.Compared with the normal group (21.00 ± 0.00) and the sham operation group (21.00 ± 0.00),the differences were all statistically significant.The rats' motor function deteriorated gradually with time after the CSCI.Osmic acid staining showed that the white matter was intact in the normal and sham groups.After being compressed the myelinated nerve fibers became swollen,degenerated and broke down.The amount of myelinated nerve fibers in the normal group,the sham operation group and the three CSCI groups was (2771 ± 108),(2675 ± 199),(2403 ± 161),(1708 ± 70) and (8 10 ± 95) respectively.The amount of myelinated nerve fibers decreased in the CSCI groups and reached a minimum on the 7th day.The difference was statistically significant.The TEM quantity analysis showed that the G-ratios in the normal,sham operation,and CSCI 1 day,3 day and7 day groups were (18.10±0.4),(17.70±1.0),(6.69 ±0.8),(5.73 ±0.4) and (4.95 ±0.5) respectively.Compared with the normal and sham operation groups,the G-ratios in the 3 CSCI groups were lower and reached their minimum on the 7th day after injury.The difference was statistically significant.TEM observation showed that the axons and myelin sheaths were intact in the normal and sham groups.After CSCI the axons became swollen and cell organelles in the axoplasm degenerated and decreased.The layers of myelin sheath shrank,folded and even wrinkled,which had an onion-like appearance.The oligodendrocytes exhibited chromatin condensation.Macrophages showed infiltration.Western blotting showed that the expression of NG2 in the CSCI groups reached a maximum on the 1st day after injury and then decreased with time.The expression of NG2 in the CSCI groups was higher than in the normal and sham groups,and the difference was statistically significant.Conclusion Demyelination occurs after CSCI-the amount of myelinated nerve fibers decreases and neurological deficits increase with time.The expression of NG2 was associated with changes in the myelin sheaths after CSCI and contributed to recovery of the myelin sheath through proliferation and differentiation to oligodendrocytes and perhaps other kinds of cells.

Objective To investigate correlation between demyelination and caspase-12 expression alteration after compressed spinal cord injury (CSCI) so as to discuss mechanism of demyelinating lesion after CSCI.Methods Seventy-five adult SD rats were randomly divided into five groups,ie,normal group,control group,compression 1 d,3 d and 7 d groups,with 15 rats per group.Models of spinal cord compression were established with a self-made device.Ultrastructure of the demyelinated nerve fibers was observed by electronic microscope and oligodendrocyte apoptosis was detected by TUNEL staining and double labeling immunofluorescence.Immunoblotting was used to defect caspase-12 that was related to cell apoptosis.Results Demyelination of nerve fiber occurred after CSCI and was aggravated with time.Apoptosis of oligodendrocytes was found after CSCI,and showed significant difference between compression 7 d group and normal group (P ＜ 0.05).Caspase-12 was also upregulated with extension of compression time.Conclusion Caspase-12 mediating oligodendrocyte apoptosis is one of the mechanisms of nerve fiber demyelination after CSCI.

Objective To observe the effects of atorvastatin on the microglia activation after traumatic brain injury (TBI). Methods Sixty adult male C57/BL6 mice were randomly divided into sham group, atorvastatin group and saline group, 20 mice for each group. The atorvastatin group and saline group were given hydraulic combat to establish TBI mouse model. The shame group underwent the same surgical procedure without being exposed to percussion injury. The atorvastatin group was treated with atorvastatin (orally, 1 mg/kg)1 h after TBI and for 7 consecutive days. The saline group was given sa?line orally. The expression of microglia (Iba-1+) at the 1st, 3rd, and 7th day after TBI and matrix metalloproteinase-9 (MMP-9) around the lesion at the 3rd day after TBI were detected by immunohistochemical staining. Tumor necrosis factor (TNF)-αwas detected by Western blot assay at the 3rd day after TBI. Results The positive expression of Iba-1+microglia was signifi?cantly decreased in atorvastatin group than that of saline group at the 1st, 3rd, and 7th day after TBI (80.00±7.44 vs. 118.40± 6.65,85.60±10.87 vs. 189.00±7.51,69.40±5.54 vs. 102.40±10.89, P<0.05). The positive expression of MMP-9 was signifi?cantly decreased in atorvastatin group compared with that of saline group at the 3rd day after TBI (86.80 ± 8.40 vs. 133.80 ± 8.46, P<0.05). Results of Western blot assay showed that the positive expression of TNF-αwas significantly decreased in astorvastatin group than that of saline group at the 3rd day after TBI (0.64±0.01 vs. 0.97±0.02,P<0.05). Conclusion Ator?vastatin can reduce inflammation factor by influencing the microglia activation after TBI in mice.

Objective: To explore the nursing experience of growth hormone provocative test in pediatric clinic. Methods: Five thousand and thirty-six children with short stature or slowing growth received combined simultaneous Levodopa Pyridostigmine stimulation test from June 2008 to October 2018 in the Child Growth Center of the First Affiliated Hospital of SUN Yat-sen University. Comprehensive nursing intervention was conducted to ensure the test carry through successfully before, during and after the test. Results: All children completed the five collections in the 120-minute growth hormone provocative test without cannula obstruction and blurt out. Some (986 out of 5 036 children, 19.58%) had different degrees of adverse reactions including nausea, vomiting, abdominal pain, sweating, salivation, dizziness, pallor, etc., most of which disappeared or alleviated after nursing, except 11 patients (0.22%) needed atropine muscular injection and 3 of whom needed intravenous fluids to release the marked symptoms. Conclusion Combined simultaneous Levodopa Pyridostigmine stimulation test is safe and practicable in pediatric clinics with nursing experience.

OBJECTIVETo explore the preoperative diagnostic value of ¹⁸F-fluorodexyglucose positron emission tomography combined with contrast enhanced computed tomography (¹⁸F-FDG PET-ceCT) in patients with colorectal cancer liver metastasis.

METHODSClinical and imaging data of 58 patients with suspicious colorectal cancer liver metastasis between April 2010 and March 2013 were retrospectively evaluated. All the patients underwent ¹⁸F-FDG PET-ceCT. On the basis of definitive diagnosis, the sensitivity, specificity, accuracy and consistency of routine PET-CT, ceCT and ¹⁸F-FDG PET-ceCT were calculated.

RESULTSA total of 147 suspicious lesions of colorectal cancer liver metastasis were found in 58 patients. Finally, 125 lesions were confinmed as malignant, of which 58 (46.4%) lesions were less than 1.0 cm. The other 22 lesions were confinmed as benign, of which 17 (77.3%) lesions were less than 1.0 cm. The diagnostic accuracy of routine PET-CT, ceCT and ¹⁸F-FDG PET-ceCT in colorectal cancer liver metastasis for the lesions more than 1.0 cm was 100%, 93.1%, 100% respectively, and the consistency with final diagnosis was perfect, moderate, and perfect respectively (Kappa value 01.00, 0.408, 1.00). For the lesions less than 1.0 cm, the accuracy was 42.7%, 78.7%, 94.7% respectively, and the consistency with definitive diagnosis was insignificance, fair, and almost perfect respectively (Kappa value -0.005, 0.305, 0.848). The area under curve(AUC) was 0.525 (95% CI: 0.407-0.462) for routine PET-CT, 0.651(95% CI:0.532-0.757) for ceCT, and 0.924 (95% CI:0.839-0.972) for ¹⁸F-FDG PET-ceCT respectively. The AUC of ¹⁸F-FDG PET-ceCT was significantly larger than that of routine PET-CT (Z=5.559, P<0.05) or ceCT (Z=4.183, P<0.05).

CONCLUSION(18)F-FDG PET-ceCT can improve the diagnostic accuracy for smaller lesions of colorectal cancer liver metastasis.

Objective To observe the value of apparent diffusion coefficient(ADC)for quantitatively evaluating progressive muscle injury of rabbit limbs in early stage of high-voltage electrical burn.Methods Twenty healthy adult rabbits were selected to establish limb high-voltage electrical burn models,which were randomly divided into 0.5,24,48 and 72 h groups(each n=5).MR diffusion weighted imaging(DWI)was collected for each group at 0.5,24,48 and 72 h after modeling,and the injured core muscles of the right hind limb and the normal muscles of the contralateral limb were taken for HE staining.The muscle's ADC,muscle fiber density(MFD)and muscle fiber diameter(D)values at the injured core of current entry and exit were compared,and those of normal muscle were also analyzed.The correlations of ADC values in injured core muscle and MFD or D values were investigated.Results There were significant differences of ADC values of injured core muscle at both the entry and exit and normal muscle,also of ADC values of injured core muscle at the entry and exit within each group(all P＜0.05).ADC values of injured core muscle at the entry and exit decreased with time going(all P＜0.05),but ADC values of normal muscle were not significantly different among different time points(P＞0.05).MFD values of injured core muscle at the entry and exit decreased with time going(all P＜0.05),while MFD values of the normal muscle,D values of the injured core muscle at the entry and exit and normal muscle were not significantly different among time points(all P＞0.05).ADC value of the injured core muscle was positively correlated with MFD value and negatively correlated with D value(rs=0.846,r=-0.507,both P＜0.05).Conclusion ADC could quantitatively evaluate the progressive muscle injury of rabbit limbs in early stage of high-voltage electrical burn.