ObjectiveTo dynamically observe the effect of N-acetylserotonin (NAS) on the expression of tumor necrosis factor-α (TNF-α) protein in retina of retinal ischemia reperfusion injury (RIRI) rats, and to explore the mechanism.MethodsBy using random number table method, 90 healthy male Sprague-Dawley rats were divided into sham operation group (n=10), RIRI group (n=40), and NAS group (n=40). The right eye was as the experimental eye. In the RIRI group and NAS group, the anterior chamber high intraocular pressure method was used to establish the RIRI model. In the NAS group, 10 mg/kg NAS was injected intraperitoneally before modeling and 30 minutes after modeling. At 6, 12, 24, 72 h after modeling, hematoxylin-eosin staining was used to observe the pathological changes of the retina, and the retinal ganglion cells (RGC) were counted. Each group was detected by immunohistochemical staining and Western blot about the relative expression of TNF-α, nuclear factor E2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) protein in the rat retina. One-way analysis of variance was used for differences between groups. The general linear regression method was used to analyze the correlation between the relative expression changes of TNF-α protein and the changes of Nrf2 and HO-1 protein expression after NAS intervention.ResultsOptical microscope observation revealed that the retinal edema of rats in the RIRI group was observed at 6, 12, and 24 h after modeling; the thickness of the retina in the NAS group was significantly thinner than that in the RIRI group, and the difference was statistically significant (F=9.645, 477.150, 2.432; P＜0.01). At 6, 12, 24, and 72 h after modeling, the retinal RGC counts in the NAS group were significantly higher than those in the RIRI group, and the difference was statistically significant (F=12.225, 12.848, 117.655, 306.394; P＜0.05). The results of immunohistochemical staining and Western blot showed that 6 h after modeling, the relative expression of TNF-α protein in the retina of the RIRI group increased significantly compared with that in the sham operation group, reaching a higher level at 12 h, and decreased at 24 and 72 h. But all were significantly higher than the sham operation group, the difference was statistically significant (immunohistochemical staining: F=105.893, 1 356.076, 434.026, 337.351; P＜0.01; Western blot: F=92.906, 534.948, 327.600, 385.324; P＜0.01). At different time points after modeling, the relative expression of TNF-α protein in the retina of the NAS group was significantly lower than that of the RIRI group (immunohistochemical staining: F=15.408, 570.482, 21.070, 13.767; P＜0.05; Western blot: F=12.618, 115.735, 13.176, 111.108; P＜0.05), but still higher than the sham operation group (immunohistochemical staining: F=40.709, 151.032, 156.321, 216.035; P＜0.01; Western blot: F=33.943, 79.729, 74.057, 64.488; P＜0.01), the difference was statistically significant; 12 h after modeling, Nrf2 in the retina of the NAS group (immunohistochemical staining: F=51.122, P＜0.05; Western blot: F=33.972, P＜0.05), HO-1 (immunohistochemical staining: F=30.750, P＜0.05; Western blot: F=18.283, P＜0.05) protein relative expression was significantly higher than that of RIRI group, and the differences were statistically significant. The results of linear regression analysis showed that the difference in the number of TNF-α+ cells in the RIRI group and the NAS group was negatively correlated with the difference in the number of Nrf2+ and HO-1+ cells (r2=0.923, 0.936; P＜0.01).ConclusionsNAS can inhibit the expression of TNF-α protein in the retina of RIRI rats and reduce RIRI. The mechanism may be related to the Nrf2/HO-1 pathway.
OBJECTIVE To determine the role of endogenous carbon monoxide(CO) in oxidant-mediated organ injury following limb ischemia-reperfusion (I/R) in rats. METHODS: Sixty-four SD rats were divided into 4 groups: Sham group, Sham + zinc protoporphyrin (ZnPP, an inhibitor of heme oxygenase activity), 2-hour ischemia followed by 4-hour reperfusion (I/R) group and I/R + ZnPP group. Carboxyhemoglobin (COHb) level in the artery blood, malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in the lung, heart, liver and kidney were detected. The 24-hour survival rate of rats was studied. RESULTS: Compared with the sham group, the COHb level and MDA content significantly increased, while the SOD activity and the survival rate significantly decreased in I/R group (P lt; 0.05). Compared with the I/R group, MDA content significantly increased, while the SOD activity, the 24-hour survival rate and COHb level significantly decreased in I/R + ZnPP group (P lt; 0.05, respectively). CONCLUSION: Limb I/R could lead to the oxidant-mediated multiple organ injury accompanied by the increase of CO level which play an important role in the defense against I/R-induced remote multiple organ injury in rats.
OBJECTIVE To testify the inductive osteogenesis of allogeneic bone matrix gelatin (BMG) in promoting intervertebral fusion. METHODS The gelatin sponge, allogeneic BMG, decalcified bone matrix (DBM) and alcohol conserved bone were implanted respectively into the intervertebral space of rabbit, whose intervertebral discs were removed before implantation. The intervertebral spaces were evaluated by X-ray and histological examination at 4, 8, and 12 weeks after operation. RESULTS No obvious immune rejection was observed. Amounts of new bone were formed in the intervertebral spaces at 4 and 8 weeks. And complete infusion of the intervertebral spaces were appeared at 12 weeks. CONCLUSION Allogeneic BMG can promote bone fusion of intervertebral spaces through osteoinduction, which suggests that allogeneic BMP is an ideal substitute for bone replacement.
ObjectiveTo investigate the effect of pulsed colloid infusion combined with continuous blood purification (CBP) for treatment of severe capillary leak syndrome (CLS).
MethodsAccording to random principle,61 patients were divided into a control group(n=21),a CBP1 group(n=18) and a CBP2 group(n=22). All patients of three groups received routine treatment according to international guidelines 2008 for management of severe sepsis. The patients in the control group also received pulsed infusion colloid combined lasix. The patients in the CBP1 and CBP2 groups also received continuous veno-venous hemofiltration(CVVH) for 72 hours. The patients in the CBP1 group received concentrated colloid infusion combined lasix,and the patients in the CBP2 group received concentrated colloid infusion combined removing fluid. Blood gas analysis and Impedance Cardiography was performed before and 24,48 and 72 hours after therapy. The angiopoietin-2(Ang-2) was measured. Also the length of ICU stay,duration of mechanical ventilation,and death rate of patients in 28 days were observed.
ResultsCompared with the control group and the CBP1 group,the length of ICU stay(days) and duration of mechanical ventilation (days) in the CBP2 group were significantly shorter(P<0.05),and the death rate in 28 days was lower(P<0.05). The patients in the CBP2 group showed more reduction in the APACHEⅡ score compared with the CBP1 group after therapy(P<0.05). The oxygenation index in the CBP2 group respectively increased at 24,48 and 72 hours after therapy(P<0.05). Compared with the control group and the CBP1 group,the oxygenation index in the CBP2 group respectively increased at the same time(P<0.05). The thoracic fluid content (TFC) in the CBP2 group respectively decreased at 24,48 and 72 hours(P<0.05) after therapy,and decreased compared with the control group and the CBP1 group at the same time(P<0.05). The serum levels of Ang-2 in the CBP2 group respectively decreased at 24,48 and 72 hours after therapy(P<0.05),and decreased compared with the control group and the CBP1 group at the same time(P<0.05).
ConclusionPulsed colloid infusion combined with continuous blood purification can reduce the severity of capillary leak and improves the outcome of patients with severe sepsis.
ObjectiveTo assess the application and the effectiveness of a strategy of combining posterior occipitocervical angle (POCA) with occipital-C2 (O-C2) angle for adjustment of occipitocervical fixation angle in posterior instrumented occipitocervical fusion.MethodsThe clinical data of 22 patients undergoing posterior instrumented occipitocervical fusions between March 2013 and January 2016 were retrospectively analysed, and all patients were performed by using a strategy combining with POCA and O-C2 angle for adjustment of occipitocervical fixation angle. All patients suffered from occipitocervical instability, including 7 males and 15 females with an average age of 44.4 years (range, 20-63 years). The patients were diagnosed as skull base depression with atlantoaxial dislocation in 20 cases and rheumatoid arthritis in 2 cases. The preoperative Japanese Orthopaedic Association (JOA) score was 13.2±2.0, and the visual analogue scale (VAS) score was 6.3±0.9. The POCA was first used to guide the pre-bending of the nail-rod system during the operation, so that POCA of 12 patients with abnormal preoperative POCA could be restored to the normal range; then intraoperative fluoroscopy was used to confirm whether the O-C2 angle was within the normal range (4 cases were abnormal and 2 cases needed intraoperative adjustment); finally, POCA and O-C2 angles were within normal range after adjustment. The postoperative complications were recorded, and the JOA and VAS scores were used to evaluate the recovery of spinal nerve function and the degree of pain relief after operation. The radiological data were collected to evaluate the bone graft fusion, the changes of postoperative POCA, O-C2 angle, and lower cervical curvature (Cobb angle).ResultsAll 22 patients were followed up 12-48 months, with an average of 24 months. No serious complications and reoperation occurred. At last follow-up, the VAS score and JOA score were 2.9±0.8 and 15.4±0.9 respectively, which were significantly improved when compared with preoperative ones (t=15.870, P=0.000; t=6.587, P=0.000). Imaging examination showed that 22 patients had occipitocervical osseous fusion, good position of internal fixator without loosening or fracture, and good occipitocervical stability. The POCA and O-C2 angles were within the normal range at 3 days after operation and at last follow-up, and there were significant differences when compared with preoperative ones (P<0.05); but no significant difference was found in POCA and O-C2 angles between at 3 days after operation and at last follow-up (P>0.05). There was no significant difference in Cobb angle of lower cervical spine between before and after operation (P>0.05).ConclusionThe strategy of combination POCA and O-C2 angle for adjustment of occipitocervical fixation angle during operation can ensure a better effectiveness.
【Abstract】 Objective To investigate the effect of verapamil on apoptosis, calcium and expressions of bcl-2 and c-myc of pancreatic cells in ischemia-reperfusion rat model. Methods Wistar rats were randomly divided into three groups: control group （n=10）; ischemia-reperfusion group （n=10）; verapamil treatment group （n=10）. The anterior mesenteric artery and the celiac artery of rats in both ischemia-reperfusion group and verapamil treatment group were occluded for 15 min followed by 12-hour reperfusion. Verapamil （1 mg／kg） was injected via caudal vein to the rats in verapamil treatment group 15 min before occlusion and 1 hour after the initiation of reperfusion, respectively; and ischemia-reperfusion group was given the same volume of salient twice intravenously. Pancreatic tissues were collected from the dead rats after twelve hours since the reperfusion. The pathologic characters of pancreatic tissue were observed under light microscope; The level of calcium in the tissue was measured by atomic absorption spectrometer; TUNEL was used to detect apoptosis of pancreatic cells; and the expressions of c-myc and bcl-2 in the cells were also analyzed by immunohistochemistry technique and flow cytometry. Results The pathologic change in verapamil treatment group was less conspicuous than that of ischemia-reperfusion group. Both the calcium level and the number of apoptotic cells in verapamil treatment group were less than those of ischemia-reperfusion group 〔（411.1±55.8） μg/g dry weight vs （470.9±31.9） μg/g dry weight, P＜0.05 and （9.5±2.9）% vs （18.4±3.1）% 〕, P＜0.05. After taking verapamil, the number of apoptotic cells decreased, whereas the expressions of bcl-2 and c-myc increased. The fluorescent indexes of bcl-2 and c-myc in verapamil treatment group were significantly higher than those of ischemia-reperfusion group (1.72±0.11 vs 1.41±0.07, P＜0.05; 1.76±0.19 vs 1.55±0.13, P＜0.05. Conclusion Ischemia-reperfusion injury can induce apoptosis of pancreatic cells. Verapamil could protect the injured pancreatic tissue by reducing the level of calcium, stimulating the expressions of bcl-2 and c-myc and inhibiting apoptosis of pancreatic cells.
ObjectiveTo investigate the effectiveness and safety of minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF) for upper lumbar disc herniation.
MethodsRetrospective analysis was made on the clinical data of 26 patients with upper lumbar disc herniation, who were in line with the selection criteria and underwent MIS-TLIF in 14 patients (MIS-TLIF group) and open transforaminal lumbar interbody fusion (OTLIF) in 12 patients (OTLIF group) between December 2007 and May 2012. There was no significant difference in gender, age, disease duration, level of disc herniation, side of disc herniation between 2 groups (P>0.05). The operation time, intraoperative blood loss, postoperative drainage volume, and complications were compared between 2 groups. The clinical outcome was assessed using the visual analogue scale (VAS) and the Oswestry disability index (ODI) scores. The fusion rate was determined by using CT three-dimensional reconstruction and dynamic lumbar radiography at last follow-up.
ResultsPrimary healing of incisions was obtained in both groups. No difference was found in operation time between 2 groups (t=0.858, P=0.399), but MIS-TLIF group had less intraoperative blood loss and postoperative drainage volume than OTLIF group (P<0.05). The average follow-up duration was 34.1 months with a range of 12-50 months. No complication of dural tear, infection, spinal nerve trauma, and implant failure occurred. The VAS scores of lower back pain and radicular pain and ODI scores at preoperation showed no significant difference between 2 groups (P>0.05). The VAS score of lower back pain and ODI score at 1 day after operation in MIS-TLIF group were significantly lower than those in the OTLIF group (P<0.05), but no difference was found in VAS scores of radicular pain between 2 groups (P>0.05). Difference in all scores was not significant at last follow-up between 2 groups (P>0.05). The fusion rate was 92.8% (13/14) in MIS-TLIF group, and was 100% (12/12) in OTLIF group at last follow-up.
ConclusionMIS-TLIF is a safe and effective procedure for upper lumbar disc herniation as an alternative to other techniques.
ObjectiveTo observe the effects of overexpression of S100A4 protein on retinal capillary cells and retinal ganglion cells (RGC) after retinal ischemia-reperfusion injury (RIRI). MethodsOne hundred healthy adult male C57BL/6 mice were randomly divided into normal control group (group C), RIRI group, adeno-associated virus (AAV2)-S100A4 green fluorescent protein (GFP) intravitreal injection group (group S), RIRI+AAV2-GFP intravitreal injection group (group GIR), and RIRI+AAV2-S100A4-GFP intravitreal injection group (group SIR), with 20 mice in each group. The RIRI model was established using the high intraocular pressure anterior chamber method in the RIRI, GIR and SIR groups of mice. Eyes were enucleated 3 days after modelling by over anaesthesia. The number of retinal capillary endothelial cells and pericytes in the retinal capillaries of mice in each group was observed by retinal trypsinised sections and hematoxylin-eosin and periodic acid-Schiff staining; immunofluorescence staining was used to observe endothelial cell, pericyte coverage and RGC survival; The relative expression of Toll-like receptor 4 (TLR4), p38 MAPK and nuclear factor erythroid 2-related factor 2 (NRF2) in retinal tissues was measured by Western blot. One-way analysis of variance was used to compare data between groups. ResultsThree days after modeling, the endothelial cell to pericyte ratio in group C was compared with group S and SIR, and the difference was not statistically significant (F=106.30, P＞0.05); the SIR group was compared with group RIRI and GIR, and the difference was statistically significant (F=106.30, P＜0.000 1). Comparison of endothelial cell coverage in each group, the difference was not statistically significant (F=3.44, P＞0.05); compared with the pericyte coverage in group C, the RIRI group and the GIR group were significantly lower, and the difference was statistically significant (F=62.69, P＜0.001). Compared with the RGC survival rate in group C, it was significantly lower in RIRI and GIR groups, and the difference was statistically significant (F=171.60, P＜0.000 1); compared with RIRI and GIR groups, the RGC survival rate in SIR group was significantly higher, and the difference was statistically significant (F=171.60, P＜0.000 1). The relative expression levels of TLR4, p38 and NRF2 proteins were statistically significant among all groups (F=42.65, 20.78, 11.55; P＜0.05). ConclusionsPericytes are more sensitive to ischemia than endothelial cells after retinal RIRI in mice, and early vascular cell loss is dominated by pericytes rather than endothelial cells. The overexpression of S100A4 protein protects against loss of pericytes and RGC after RIRI by inhibiting the TLR4/p38/NRF2 signaling pathway.
Objective To evaluate the effect of regiono-perfusional chemotherapy of pancreatic adenocarcinoma, and to seek the management of its complications. MethodsThirty-six patients with unresectable pancreatic adenocarcinoma received selectively intra-arterial catheterization and perfused with 5-Fu, ADM, DDP. Results Six patients had complete response, 15 partial response, and one underwent radical resection subsequently. Cmplications occurred in 14 patients with 2 patients died of complications.Conclusion Regiono-perfusional chemotherapy of pancreatic adenocarcinoma is effective, but the complications can not be neglected.
Objective
To evaluate the clinical effects of surgical treatment for 30 patients with discogenic low back pain.
Methods
A total of 30 patients with 36 intervertebral discs were treated with posterior approach lumbar discectomy and interbody fusion with internal fixation by strict criteria. All patients were followed up for one year. The low back pain before and one year after surgery was evaluated by Japanese Orthopaedic Association (JOA) score and Visual Analogue Scale (VAS) score.
Results
The patients’ JOA score increased from 14.6±2.3 (before operation) to 27.1±0.9 (one year after operation) (t=–26.936, P<0.001), while the patients’ VAS score decreased from 6.2±1.6 (before operation) to 1.4±0.9 (one year after operation) (t=16.335, P<0.001), and the differences were significant.
Conclusion
When the conservative treatment is invalid, the operation of posterior lumbar intervertebral fusion is an effective method for the patinets with discogenic low back pain.
