Objective:To investigate the protective effects and mechanism of exogenous milk fat globule-epidermal growth factor 8(MFG-E8) on intestinal injury and ferroptosis in mice with acute pancreatitis (AP) and its mechanism.Methods:A total of 24 male C57 BL/6 mice were randomly divided into the normal control group, AP group, AP+ MFG-E8 group (MFG-E8 group), and AP+ MFG-E8+ cilengitide group (cilengitide group), with 6 mice in each group according to the random number table. The mice of normal control group were intraperitoneally injected with 0.9% sodium chloride solution. In the AP group, MFG-E8 group, and cilengitide group, the mice were intraperitoneally injected with 8% L-arginine twice at 1-hour intervals to induce the AP model. In the MFG-E8 group, mice were intraperitoneally injected with 20 g/kg of MFG-E8 at 2 hours after L-arginine injection. In the cilengitide group, mice were intraperitoneally injected with 20 mg/kg of cilengitide at 1 hour after the L-arginine injection, and 20 g/kg of MFG-E8 1 hour later. The mice were sacrificed and blood samples and intestinal tissues were collected at 72 hours after the first L-arginine injection. Hematoxylin-eosin staining was used to evaluate intestinal tissue injury. Myeloperoxidase (MPO) immunofluorescence staining was performed to detect neutrophils in intestinal tissues.Adenosinetriphosphate (ATP) levels were examined to detect changes in mitochondrial function. Malondialdehyde (MDA) and superoxide dismutase (SOD) levels were tested to check the level of intestinal oxidative stress. Dihydroethidium (DHE) fluorescent probe was used to label the oxygen free radicals in intestinal tissues. The expression of glutathione peroxidase 4 (GSH-Px4), solute carrier family 7 member 11 (also named xCT), and ferroptosos suppressor protein-1 (FSP-1) in intestinal tissues were detected by western blotting. Indepent-samples t-test, one-way ANOVA, and Student-Newman-Keuls test were performed for statistical analysis. Results:Intestinal tissue injury and inflammatory cell infiltration in mice were induced by intraperitoneal injection of L-arginine. Compared with those of AP model group, the intestinal pathology score, MPO fluorescence quantification and DHE fluorescence density of MFG-E8 group were significantly decreased (3.93±0.57 vs. 1.73±0.74, (26.33±4.49)/field vs. (11.00±3.27)/field, (39.67±5.79)/field vs. (12.33±3.68)/field), while the contents of ATP, MDA and SOD were increased ((77.09±8.52) μmol/g vs. (119.87±6.83) μmol/g, (0.10±0.01) μmol/g vs. (0.17±0.02) μmol/g, (105.67±6.93) U/mg vs. (144.49±18.55) U/mg), and the differences were statistically significant ( t=3.33, 3.93, 5.63, 8.77, 6.54, 4.38; all P<0.05). The results of Western blotting showed that GSH-Px4, xCT, and FSP-1 in the intestinal tissue of AP mice in the MFG-E8 group were all elevated compared with those of AP group, the relative expression levels were 1.22±0.19 vs. 0.55±0.09, 1.48±0.12 vs. 0.34±0.08, and 0.48±0.08 vs. 0.04±0.03, and the differences were statistically significant ( t=5.60, 14.39, 9.53; all P<0.05). Intraperitoneal injection of integrin αVβ3 receptor inhibitor cilengitide effectively antagonized the protective effects of MFG-E8 on intestinal injury in AP mice. Compared with MFG-E8 group, the histopathological score, MPO quantification and DHE fluorescence density of cilengitide group (3.53±0.50, (27.67±6.02)/field, and (31.33±3.86)/field, respectively) all increased, the expression of ATP, MDA and SOD were inhibited ((77.41±8.51) μmol/g, (0.19±0.04) mol/g, (100.46±8.15) U/mg); and GSH-Px4, xCT and FSP-1 all decreased, with the relative expression levels of 0.59±0.11, 0.16±0.06, and 0.10±0.03, respectively, and the differences were statistically significant ( t=3.02, 3.44, 5.04, 8.70, 4.01, 4.86, 5.05, 17.47, 8.34; all P<0.05). Conclusion:MFG-E8 alleviates intestinal oxidative stress and ferroptosis by integrin αVβ3 receptor, thereby reducing intestinal injury and inflammation in AP mice.

Objective:To investigate the protective effects and mechanism of exogenous milk fat globule-epidermal growth factor 8(MFG-E8) on intestinal injury and ferroptosis in mice with acute pancreatitis (AP) and its mechanism.Methods:A total of 24 male C57 BL/6 mice were randomly divided into the normal control group, AP group, AP+ MFG-E8 group (MFG-E8 group), and AP+ MFG-E8+ cilengitide group (cilengitide group), with 6 mice in each group according to the random number table. The mice of normal control group were intraperitoneally injected with 0.9% sodium chloride solution. In the AP group, MFG-E8 group, and cilengitide group, the mice were intraperitoneally injected with 8% L-arginine twice at 1-hour intervals to induce the AP model. In the MFG-E8 group, mice were intraperitoneally injected with 20 g/kg of MFG-E8 at 2 hours after L-arginine injection. In the cilengitide group, mice were intraperitoneally injected with 20 mg/kg of cilengitide at 1 hour after the L-arginine injection, and 20 g/kg of MFG-E8 1 hour later. The mice were sacrificed and blood samples and intestinal tissues were collected at 72 hours after the first L-arginine injection. Hematoxylin-eosin staining was used to evaluate intestinal tissue injury. Myeloperoxidase (MPO) immunofluorescence staining was performed to detect neutrophils in intestinal tissues.Adenosinetriphosphate (ATP) levels were examined to detect changes in mitochondrial function. Malondialdehyde (MDA) and superoxide dismutase (SOD) levels were tested to check the level of intestinal oxidative stress. Dihydroethidium (DHE) fluorescent probe was used to label the oxygen free radicals in intestinal tissues. The expression of glutathione peroxidase 4 (GSH-Px4), solute carrier family 7 member 11 (also named xCT), and ferroptosos suppressor protein-1 (FSP-1) in intestinal tissues were detected by western blotting. Indepent-samples t-test, one-way ANOVA, and Student-Newman-Keuls test were performed for statistical analysis. Results:Intestinal tissue injury and inflammatory cell infiltration in mice were induced by intraperitoneal injection of L-arginine. Compared with those of AP model group, the intestinal pathology score, MPO fluorescence quantification and DHE fluorescence density of MFG-E8 group were significantly decreased (3.93±0.57 vs. 1.73±0.74, (26.33±4.49)/field vs. (11.00±3.27)/field, (39.67±5.79)/field vs. (12.33±3.68)/field), while the contents of ATP, MDA and SOD were increased ((77.09±8.52) μmol/g vs. (119.87±6.83) μmol/g, (0.10±0.01) μmol/g vs. (0.17±0.02) μmol/g, (105.67±6.93) U/mg vs. (144.49±18.55) U/mg), and the differences were statistically significant ( t=3.33, 3.93, 5.63, 8.77, 6.54, 4.38; all P<0.05). The results of Western blotting showed that GSH-Px4, xCT, and FSP-1 in the intestinal tissue of AP mice in the MFG-E8 group were all elevated compared with those of AP group, the relative expression levels were 1.22±0.19 vs. 0.55±0.09, 1.48±0.12 vs. 0.34±0.08, and 0.48±0.08 vs. 0.04±0.03, and the differences were statistically significant ( t=5.60, 14.39, 9.53; all P<0.05). Intraperitoneal injection of integrin αVβ3 receptor inhibitor cilengitide effectively antagonized the protective effects of MFG-E8 on intestinal injury in AP mice. Compared with MFG-E8 group, the histopathological score, MPO quantification and DHE fluorescence density of cilengitide group (3.53±0.50, (27.67±6.02)/field, and (31.33±3.86)/field, respectively) all increased, the expression of ATP, MDA and SOD were inhibited ((77.41±8.51) μmol/g, (0.19±0.04) mol/g, (100.46±8.15) U/mg); and GSH-Px4, xCT and FSP-1 all decreased, with the relative expression levels of 0.59±0.11, 0.16±0.06, and 0.10±0.03, respectively, and the differences were statistically significant ( t=3.02, 3.44, 5.04, 8.70, 4.01, 4.86, 5.05, 17.47, 8.34; all P<0.05). Conclusion:MFG-E8 alleviates intestinal oxidative stress and ferroptosis by integrin αVβ3 receptor, thereby reducing intestinal injury and inflammation in AP mice.

Objective To analyze the clinical characteristics of Brucellar spondylitis and evaluate its surgical treatment effect.Methods From 2002 to 2015,the clinical data of diagnosed patients with Brucellar spondylitis aged ≥65 years old were collected retrospectively in the First Affiliated Hospital of Hebei North University,patients' clinical manifestations and characteristics,laboratory diagnosis,imaging changes and surgical treatment effects were analyzed.Pain and imaging scores and clinical efficacy were evaluated according to follow-up data within 12 months after surgery.Results A total of 38 cases of senile patients with Brucellar spondylitis were studied.The main clinical symptoms during the visit were persistent severe lumbar back pain,local tenderness,obvious percussion pain,muscle spasm,and constrained movements of spine.Rose bengal plate test (RBPT) was positive in 11 cases,serum tube agglutination test (SAT) titer was higher than 1:100 in 28 cases.During the operation,28 cases of patients with inflammatory granuloma or abscess were cultured positive for Brucella.Before surgery,manifestations of X-ray:of which 29 cases showed intervertebral space narrowing,the density increased,and the margins of the verbebral bodies destructed;of which 9 cases showed vertebral body bone sclerosing hyperplasia in the shape of a bird's beak,forming a bone bridge with the adjacent vertebral body margin.Manifestations of CT:of which 29 cases showed different sizes and multiple lesions of the vertebral body margin,and hyperplasia and sclerosis around the lesion,and destruction of the lesion in the new bone tissue to form "lacy vertebra";in 9 cases,the destruction of intervertebral discs showed isodensity shadow,and the hyperplasia and sclerosis of articular surface resulted in the formation of "labial" osteophytes;of which 10 cases showed vertebral body destroyed the plane in which the bilateral psoas were widened and abscess formed.Manifestations of MRI:of which 38 cases showed that the vertebral bodies,intervertebral discs,accompaniment and the intra of vertebral canal were uneven high signal and dural sac or cauda equina compression.The pain scores [(2.1 ± 0.2),(0.7 ± 0.4),(0.2 ± 0.1),(0.0 ± 0.0),(0.0 ± 0.0) scores] at 2 weeks and 1,3,6,12 months after surgery were significantly lower than that before surgery [(9.2 ± 0.3) scores,P ＜ 0.05].The blind imaging evaluation scores [(4.68 ± 0.04),(4.92 ± 0.08),(5.00 ±0.00) scores] at 3,6 and 12 months after surgery were significantly higher than that before surgery [(0.37 ± 0.03) scores,P ＜ 0.05].The cure rates of clinical efficacy [92.11％ (35/38),100.00％ (38/38)] at 6 and 12 months after surgery were significantly higher than that of 3 months after surgery [78.95％ (30/38),P ＜ 0.05)].Conclusions Senile Brucellar spondylitis has the typical imaging features,laboratory examination is helpful to early diagnosis.Surgery is much better to relieve the pain and stable spine function and accelerate rehabilitation.

BACKGROUND:Foreign scholars have obtained a success to cure fracture by implanting the complex of red bone marrow and formation factor.Due to the in vitro culture process is not necessary,the complex of red bona marrow and scaffold formation factor is only required to be implant immediately,called uncellular tissue engineered bone.OBJECTIVE:This study innovatively constructs uncellular tissue engineered bone with autologous red bone marrow wrapped by fascial flap with pedicle,and validates the superiority of repairing bone defects.DESIGN,TIME AND SETTING:Homebody controlled animal experiment was performed in the Hebei North University and the Experiment Center of the Affiliated Hospital to Hebei North University from December 2007 to February 2009.MATERIALS:A total of 24 News Zealand albino rabbits,aged 4-5 months; uncellular tissue engineered bone was a mixture of autologous red bone marrow and osteoinductive absorbing materials containing bone morphogenetic proteins.METHODS:Bone defect models were induced on adult New Zealand rabbits' right radial bone,left side served as control group,only implanted with osteoinductive absorbing materials complex,while right side served as experiment group,which contained fascial flap with pedicle.A fascial flap prepared with capillary network containing nameless blood vessel pedicle was located to be adjacent to the bone defect using micro-surgical technique,to wrap the tissue engineered bone and to fill the bone defect.MAIN OUTCOME MEASURES:At 4,8,12,16 weeks postoperation,six rabbits were tested by radiograph,spectrodensitometry,gross morphology observation,histological inspection,quantitative analysis of bone morphometry in bone defect area and analysis of vessels image in the junctional zone.RESULTS:①X-ray determination:At 16 weeks,the implant surrounding bone defects formed bone shaft structure in the control group,cortical bone was not continuous and medullary cavity was obstructed; in the experiment group,normal bone shaft structure was formed and recanalization of medullary cavity was observed.②Histological observation:At 16 weeks,few vessels grew into implant in the control group,mature bone trabecular were observed,and medullary cavity was obstructed; in the experiment group,the implanting materials were completely degraded and substituted by new bone,mature bone structure formed and recanalization of medullary cavity was observed.③Quantitative analysis of bone morphometry in bone defect area:At 4,8,12,16 weeks postoperation,the volume of bone trabecula in the experiment group was more than that in control group (P ＜ 0.05).④Analysis of vessels image in the junctional zone:The area of vessels in the unit area in the experiment group was greater than that in control group in every time stage (P＜0.05).CONCLUSION:The uncellular tissue engineering complex constructed by autologous red bone marrow wrapped by fascial flap with pedicle shows double effects of hymeno-inducing regeneration of bone and the vascularization.It is feasible to repair large-segment bone defects.It has obvious therapeutic effect in the aspects such as reducing the bone defect reparation time and advancing the quantity and quality of the bone generation.

Objective To study the effect of fascial flap with vessels inducing the vascularization of uncel-lular tissue engingeering complex and the regenration of bone on the repair of bone defect, so as to provide the basis for the clinical application. Methods An animal model of bone defect on adult Newzland rabbits'right radial bone was established .and autologous red bone marrow were taken out and mixed into uncellulax tissue engineering comple-xes with OAM which contained BMP. The experiment animals were divided into two groups : experiment group and control group( n = 12 for each ). The control group was only implanted with complexes, meanwhile, the experiment group had fascial flap with vessels. By microsurgery technology,a non-named fascial flap with vessels was prepared, which belonged to capillary net,around the bone defect,and let it wrap tissue engineering complex,fill up bone de-fect. In a certian time, radiograph(X-ray) and light density measure was conducted, gross morphology and histological inspection was exmained. Bone shape measurement analysis and image of vessel analysis were conducted. All the sta-tistics were analyzed by the SPSS 11.5 software. Results Because of mechanically preventing fiber connective tis-sues and surrounding soft tissues from entering the areas of bone defect by fascial flap, it can keep bone defect having a relative stable environment ;The subfascial space itself, and also the shape and mass of filled-in subject had the de-cisive effect on the results of the regeneration of the bone; Owing to the establishment of blood supply during the con-structing tissue engineering complex. The experiment group was obviously superior to the control group. Compared with control group,the absor bance obviously increased in experiment group [(0. 732 ± 0. 021 ) vs (0. 651± 0.018)] (P < 0. 001 ) four weeks after the operation; also the bone trabecular body was significantly increased [(2.32±2.57)% vs(19.37±3.52)% ,(8.37±3.52)% vs(30.24±3.42)% ,(28.57±2.98)% vs(58.76± 4.62)% ,(47.24±3.42)% vs(88.72±5.84)%] ,and capillary area [(5.04±1.62)% vs(17.53±2.86)%, (10.37 ±2.96)% vs(35.24±1. 13)%,(18.20±2. 12)% vs(48.76±4. 62)%,(17.82 ±2. 74)% vs (57.72 ±5.84)%] (P <0.05) at each time period(4 weeks,8 weeks,12 weeks,and 16 weeks after operation). Despite of growth of implant's internal vessel, the number and speed of forming bone trabecula and cartilaginous tis-sue, even developing of mature bone structure, recreating of diaphysis structure, reconstructing of marrow cavity, ab-sorbing and decomposing of implant, the experiment group was obviously superior to the control group. Conclusions The induction of fascial flap with vessels shows double effects, one of which is the vascularization of uncellular tis-sue engineering complex and the other is membrane guided bone regeneration, So the method has a wonderful effect on the repair of bone defect.

Objective To study the relationship of proliferation and activation of T lymphocyte subsets and disease progression in antiretroviral-naive human immunodeficiency virus(HIV)-1-infected individuals.Methods Forty-nine antiretroviral-naive,chronically HIV-1 infected patients and 16 healthy,HIV-1 negative controls were enrolled in this study.The patients were divided into 3 groups according to their CD4+T cell counts:<200×106/L,(200-350)×106/L and>350×106/L.Peripheral blood mononuclear cells(PBMC)were isolated.T cell proliferation index was measured by Ki-67 staining.T cell activation was detected by CD38 staining.The samples were analyzed by flow cytometry.The data were compared by one-way ANOVA.Results The percentage of Ki-67+cells in CIM+T ceils was 7.92%±4.37%in CD4+T cell<200×106/L group,which was significantly higher than those 0.39%d:0.24%in control group,2.61%±2.12%in(200-350)×106/k group and 2.65%±2.13%in>350 X106/L group(F=21.961,P<0.01).The percentage of Ki-67+cells in CD8+T ceils in CD4+T cells<200×106/L group was 2.87%±1.13%,which was also much higher than those in other 3 groups(0.15%±0.90%,1.40%±1.17%,1.22%±0.80%,respectively F=19.203,P<0.01).The Ki-67'CD4'T cells and Ki-67+CD8+T cells were inversely correlated with CD4+T cell counts(r=-0.654,r=-0.539,respectively;P350×106/L group(F=14.333,F=15.412,respectively;P<0.01).The expressions of Ki-67+ on CD4+ and CD8+T cells were positively correlated with CD38 expression(r=0.527,r=0.391,respectively;P=0.002).Conclusions The proliferation and activation of T lymphoeytes are enhanced during HIV/AIDS disease progression.And T eell activation may be the result of persistent immune activation.

[Objective]To analyze the clinical diagnosis and therapic level,the clinical manifestation,image manifestation and surgical treatment of the sacral epidural cystis.[Method]Eighteen cases with the sacral epidural cystis were analyzed retrospectively,17 cases(95%)with dull pain of the lumbosacral area,16 cases(90%)with radiated pain of the lower limbs and intermittent limping,14 cases(77%)with functional lesion of the nerves,2 cases(11%)with tumor of pelvic,cavity or hypogastrium.All of them were taken AP views of X-ray,17 cases taken two-dimentional CT scannings,11 cases taken MRI,7 cases taken myelography and 4 cases taken CTM scannings.[Result]The rate of the error diagnosis was 78% in the clinic,while the success diagnosis rate was 17% in the X-ray,72% in the CT,100% in the MRI,myelography and CTM.The exairesis were done in all the 18 cases,and the epidural cystis were diagnosed in pathology.No reoccurrance was found at 15~32 months follow-up.[Conclusion]There are characteristic changes in the imageology and clinic of the sacral epidural cystis.Using minimall invasive surgery(laparoscope vertebroscope and epiduroscopy)has no confirm result till now.Microsurgical techniques contribute to the improvement of the total removal rates for the intraspinal tumors.So operation is reliable method to treat this disease and to prevent its recurring.

[Objective]The naming,pathogeny,clinic,patho logical character and image manifestation of the parosteal lipoma were discussed in this paper combined with the literatures.[Method]Combined the 20 cases reported in the internal literatures,the photographs have been taken in all the 34 cases,CT in the 18 cases,MRI in the 4 cases.[Result]The data demonstrated that this disease betided in any age,more in the adult,developed more in the long canal bone of the limb,and developed in the flat bone(pelvis,scapula,costal bone,patella),anomaly bone(thoracic verthbrae,lumbar,ansa sacralis),cauda bone,short canal bone and instep bone secondly.The rate of the error diagnosis were 40.91% in the clinic,27.27% in the X-ray and no one in the CT and MRI.The excision was done in all the 34 cases,and lipoma were diagnosed in pathology,3 cases were malignant slightly.Followed up for 2 to 10 years,only 1 cases recurring and malignant.[Conclusion]The name of the parosteal lipoma is optimal,the pathogeny is not clear,and related to the heredity,hurt and inflamation.There is lipoma with bone pedicel in pathology;and adipose tissuse in cytology.There are characteristic changes in the X-ray,CT and MRI.

[Objective] To improve the clinical diagnosis and therapic effect and to discuss the clinical and imageologic manifestations and treatment of the brucellosis spondylitis(BS).[Method]Sixteen patients with BS were all taken radiographs and CT scanning and seven cases were also taken MRI.The test of SAT,test of RBP and ELISA were used in accessing BS.Pathogenic bacteria were examined in 11 cases.All cases were treated with anti-brucellosis agents and other conservative measures.Seven cases were followed with a focal debridement and 2 cases withminimal invasive surgery.[Result]Fourteen cases were misdiagnosed as spine TB before they were hospitalized with rate of error diagnosis 87.5%,even careful clinic and radiographic examination,and CT scanning had been taken,and 5 cases were misdiagnosed by MRI with the misdiagnosis rate 71.4%.The BS occurred far more in the lumbar vertebrae than in the thoracic vertebrae with its highest morbidity at the L4.Among 16 cases,the titers of SAT test were all beyond 1.160,the test of RBP all showed positive and IgG,IgM showed positive by ELISA.Eleven cases was confirmed BS diagnosis by phthogenic bacteria examination.Only 12 cases were followed up for 1～2 years(including 8 surgical-treated cases).Ten cases were cured without recurrence,and 2 cases improved.[Conclusion]Brucellosis spondylitis commonly misdiagnose before hospitalization.Characteristic imageologic changes and clinical appearance,combined tests of SAT and RBP are useful for diagnosis.Long-term and enough amount sensitive antibiotics and other conservative measures is still the major and reliable measures in treatment of BS and prevent recurrence,additional minimal invasive surgery or focal debridement could shorten the course of therapy,increase the cure rate and decrease its complication.