0.05).Conclusions The TNF-?levels differed among the sta- ges of disease and MS subtypes,but no marked relation existed between the TNF-?level and the EDSS score.

Objective:To investigate establishing the multiple clinical processes model and pathological characteristics of EAE model,and to provide the experimental value for reseaching MS. Methods:The animal model was established in Wistar rat by immunization with complete Freund adjuvant and GPSCH, the pathological changes of EAE were studied with the aid of light microscopy. Results : We can divide the EAE into five types by pathological changes and clinical manifestation: acute EAE, relapsing-remitting EAE, persistent progress EAE,benigh form EAE and delitescence EAE. Every type mainly showed congestion and inflammatory cuff of small blood vessels,disintegration and disappear of myelin sheath and degeneration of neurons,especially in spinal cord. Conclusion:Multiple clinical processes on EAE model of Wistar rat were established for the first time.it is an ideal animal model in studying MS.

Objective:To investigate pathological changes and the expression of the matrix metalloproteinases in rat EAE.Methods:The pathological changes of EAE were studied in Wistar rat with the aid of light and electron microscope and the expression and distribution of MMP-2,MMP-9 in different tissues were detected with the method of immunohistochemistry.Results:Light microscopy showed inflammatory cuff around small blood vessels were evident, disintegration and disappear of myelin sheath were observed. Electron microscopy showed a lot of loose and fragmental spiremes of myelin sheath, the component of axons and nissl bodies in neurons were disappeared. MMP-2,-9 were expressed intensively in vascular endothelial cells, meninges and accumulative inflammatory cells.Conclusion:MMPs take the roles in every aspect of the pathological changes in EAE, it can destroy the blood brain-barrier, degrade the myeline sheath, damage the axons and generate immunogens.

BACKGROUND: Glial cell line-derived neurotrophic factor (GDNF) ischaracterized by its trophic function on motor neurons, but there is stilllack of quantitative data concerning the influence of different concentra tions of the neurotrophic factor on the growth of in vitro cultured motorneurons. OBJECTIVE: To observe the influence of GDNF on neuronal growth byobserving fetus rat spinal cord motor neurons cultured in vitro. DESIGN: Verifying observation taking in vitro cultured cells as subjects. SETTING: Neurological Department of the Second Hospital Affiliated toHebei Medical College. MATERIALS: This experiment was conducted in the laboratory of Neu rological Department, the Second Hospital Affiliated to Hebei Medical College, between January 2001 and September 2002. Adult male and female rats were raised together in the same cage, embryonic rats at 15 days of gestation were obtained for spinal cord separation. METHODS: Ventral spinal tissues were obtained from embryonic rats at 15 days of gestation for prinary in vitro culture. They were divided into four groups according to the density of GDNF, namely 1, 10, 50 and 100 μg/L GDNF groups, while the culture medium in control group did not contain GDNF. Neurons were cultured in 8 wells foreach group, which was repeated for two batches. Then the influence of GDNF on spinal cord motor neu rons was observed from the perspective of cell morphology with MTF method. MAIN OUTCOME MEASURES: The survival rate of motor neurons andthe length of cell processes. RESULTS: ① The length of spinal cord motor neuronal processes: It was found obviously longer in GDNF 1 μg/L group, 10 μg/L group, 50 μg/L group and 100 μg/L group than in control group [(107.4±35.406 8,160.5±38.564 9, 450.5±60.640 3, 293.5±67.381 4, 82.8±7.972 5) μm, t=2.610-2.647, P ＜ 0.01]. ② Cell survival rate: It was higher in GDNF 1 μg/L group, 10 μg/L group, 50 μg/L group and 100 μ g/L group than in control group [(13.9±0.899 9, 16.1±0.668 0, 20.1±0.667 9, 26.0±0.603 0,10.5±0.782 0) μm, t=2.211-2.312, P ＜ 0.05]. ③ MTT colorimetric analy sis: It was obviously higher in GDNF 1 μg/L group, 10 μg/L group, 50 μg/Lgroup and 100 μ g/L group than in control group [(0.350±0.059 8, 0.366 7±0.071 9, 0.381 9±0.063 8, 0.395 3±0.060 5, 0.285 8±0.032 5) μm,t=2.259-2.577, P ＜ 0.05]. CONCLUSION: GDNF of different concentrations exerts different effects on in vitro cultured embryonic spinal cord motor neurons.

Objective To establish an experimental autoimmune encephalomyelitis (EAE) model in Wistar rats with myelin basic protein fragment (MBP69-85) and observe its pathological changes. Methods MBP69-85 dissolved in normal saline was mixed with complete Freund's adjuvant (CFA) (including 6 mg bacille Calmette Guerin) to prepare the encephalitogenic emulsion. Ten out of 70 Wistar rats were chosen as a control group, the others were divided equally into group A,B,C according to the difference of the encephalitogenic emulsion. Rats in group A were immunized with 50 μg MBP69-85 +CFA (including 6 mg BCG). Rats in group B were immunized with 25 μg MBP69-85+CFA (including 6 mg BCG). Rats in group C were immunized with 25 μg MBP69-85+CFA (including 12 mg BCG). The pathological changes of brain and spinal cord tissues were examined by light microscopy after HE staining and immunohistochemistry of MBP and NF. Results Some of the Wistar rats immunized with 50 μg MBP69-85 showed disorder at 12～16 days after immunization. The clinical symptoms included tail acratia or paralysis of tail and limbs, head tilt, etc. and the mean score was 2.38±1.89. There were infiltration of inflammatory cells inside nervous tissue and perivascular cuffings in HE stained sections. The immunohistochemistry of MBP and NF showed demyelination in the white matter and axon injury. Conclusion To some extent, the establishment of EAE depends on the dose of the immunizing antigen. The BCG in CFA was not the major cause of morbility of the rats. The EAE model induced with MBP69-85 in Wistar rats, showing typical clinical symptoms and pathological changes of multiple sclerosis, is a reliable animal model for the study of pathogenesis and treatment of multiple sclerosis.

Objective:EAE model of male and female Wistar rats was established respectively.The sex difference was evaluated.Methods:40 wistar rats were devided into male and female group (n=20).EAE model was established respectively in the two groups using Guinea pig spinal cord homogenate without PBS perfusion.30 days later,the spinal cord were taken and embedded in wax,and paraffin sections of the two groups were examined by histopathology.The differences in onset time,incidence rate,course of disease and neurologic score between the two groups were evaluated.Results:The onset time of female group was 13.67?3.50 days,its incidence rate was 60%.The onset time of male Wistar rat was 12.18?1.55 days,and its incidence rate was 85%.The feature of diseases course in female rat was relapse after recovery,but that in male rat was transient.The average clinical score of female group was 2.20?1.96,and that of male group was 3.46?1.61.Conclusion:Compared with female Wistar rat,the male had higher incidence rate and its diseases course was transient.The results of our study provided a useful foundation for further studies on the pathogenesis and the sex differences of multiple sclerosis.

Objective: To explore the modified surgical method of supra- and infratentorial epidural hematoma (SIEDH) and analyze the related anatomy of the occipital bone. Methods: CT scan data of 30 adult patients with no history of craniocerebral trauma were collected in our hospital from January to August 2019. The median sagittal plane was taken to define line A (from lambdoid suture to the extraoccipital turbercle) and line B (from extraoccipital turbercle to the posterior edge of the foramen magmun). The angle θ was defined between the two lines. The mean value of θ was analyzed for comparison between genders and for exploration of the anatomical basis of single supratentorial craniotomy for SIEDH. Eight patients with SIEDH were recruited in the same period to analyze the procedure and results of the modified surgical method. Results: The average angle of the θ was (117.4±4.3)° for men and (130.0 ±4.9)° for women; the minimum was 108.3° for men and the maximum was 138.7 ° for women. The θ was smaller in men than in women (P<0.001). The smaller the angle was, the more conductive it was to detect the lesions from superior to inferior tentorial apartment. The bone flaps of the eight patients were designed above the transverse sinus. During the operation, epidural hematoma was completely removed, and no skull defects occurred like in conventional supra-inferior tentorial craniotomy. Conclusion: The supratentorial craniotomy above the tranverse sinus has reliable anatomical basis and is an effective surgical method for SIEDH.

Objective To explore the ultrasonographic features of the gallbladder with malaria in stage of attack (MSA) at Sudan.Methods A total of 67 cases of MSA patients were divided into common type of malaria (CM) and sever malaria (SM).All patients underwent routine abdominal ultrasound examination and the ultrasonographic features of the gallbladder were divided into type Ⅰ,Ⅱ and Ⅲ according to the volume of gallbladder,thickness of the gallbladder wall and chole stasis.The relationship between clinical types and the ultrasonic types were analyzed.Then the differences of liver function among normal,Ⅰ,Ⅱ and Ⅲ types were compared statistically.Results In all of the 67 MSA patients,52 cases were CM and 15 cases were SM.The gallbladder ultrasonography showed that normal gallbladder were in 14 cases,type Ⅰ in 28 ca ses,type Ⅱ in 19 cases,type Ⅲ in 6 cases.The ultrasonic types of the gallbladder had significant difference in SM patients (type Ⅰ 3 cases,type Ⅱ 7 cases,type lⅢ 5 cases) and CM patients (normal 14 cases,type Ⅰ 25 cases,type Ⅱ 12 cases,type Ⅲ 1 case;P＜0.001).Ultrasonic types of the gallbladder had no difference between Chinese and Sudanese (P＞ 0.05).There were statistical significant differences of liver function among normal,Ⅰ,Ⅱ and Ⅲ types of the gallbladder (P＜0.001).Conclusion Most of the MSA patients have ultrasonic changes of gallbladder.And there are more serious pathological changes of gallbladder in SM than that in CM.

BACKGROUND:It has been found that central nervous system is involved in Guillain-Barre syndrome and Miller-Fisher syndrome, and the involved sites include optic nerve, brain stem and cerebellum. Abnormal signal of MRI can be observed in the brainstem and spinocerebellar tract of patients with Miller-Fisher syndrome. To establish an animal model of encephalitis after infection of Campylobacter jejuni, and investigate the mechanism of formation by means of imaging, immunology and pathology.OBJECTIVE: To construct an animal model of lesion of central nervous system after infection of Campylobacter jejuni Penner 4.DESIGN: A randomized grouping designed, controlled animal experiment.SETTING: Department of Imaging and Department of Neurology, Second Hospital of Hebei Medical University.MATERIALS: The experiment was carried out in the Laboratory of Molecular Imaging, the Second Hospital of Hebei Medical University between August and December 2003. Fifteen healthy flap-eared rabbits were randomly divided into experimental group (n=10) and control group (n=5).METHODS: In the experimental group, Campylobacter jejuni inactivated bacteria liquor was completely emulsified with complete Freund adjuvant (CFA) of the same volume in week 1, and then the rabbits were immunized with subcutaneous injection at multiple points of bilateral axilla, bilateral groins and side of back spine, 1 mL for each site, and 5 mL for each rabbit; The rabbits were further immunized with intraperitoneal injection of simple Campylobacterjejuni inactivated bacteria liquor in the following every two weeks, 5 mL for each time in each rabbit for 5 times. In the control group, the Campylobacter jejuni inactivated bacteria liquor was replaced by saline of the same volume, the injected method and time were all the same as those in the experimental group. Evaluative methods: ①Symptoms and physical signs: their mental status, conditions of diet, urine and excrement, and activities of limbs were observed; ② Serological examination: the contents of anti-Campylobacterjejuni antibody, anti-IgG GM1 antibody and myelin basic protein (MBP) were detected with enzyme-linked immunoadsorbent assay (ELISA); ③ MRI examination was applied to the randomly selected rabbits before every immunization with Toshiba 1.5 T MRI instrument. The scanning sequence included spin-echo T1-weighted image with the scanning parameter of 500/15 ms (TR/TE); rapid spin-echo T2-weighted image, 4 000/108 ms (TR/TE); fluid attented inversion recovery (Flair) sequence, the parameter was 10 000/120 ms (TR/TE), inversion angle was 90°. The thickness of scanning layer was 4.0 mm, and the layer space was 0.8 mm. ④ Histological examination: At 4 weeks after the first immunization, the attacked animals were induced to death by cardiac perfusion, and the skull was opened immediately to remove optic nerve, part white matter, hippocampus, brainstem, cerebellum and spinal cords of neck, chest and waist, which were fixed with formaldehyde solution (40 g/L),and hematoxylin-eosin (HE) staining, fast blue staining and MBP immunohistochemical staining were performed respectively. At 10 weeks after immunization, 5 randomly selected rabbits in the experimental group and the 5 rabbits in the control group were treated with the same methods to obtain the histological samples.MAIN OUTCOME MEASURES: The symptoms and physical signs,contents of anti-Campylobacterjejuni antibody, anti-IgG GM1 antibody and MBP, imaging observation and histological examination were mainly observed.RESULTS: Fifteen animals were enrolled, 14 were involved in the analysis of results, 1 rabbit in the experimental group died at 4 weeks after immunization. ① Mental symptoms and disorder of limb's activity occurred in 1 rabbit in the experimental group at 2 weeks after immunization. ② In the experimental group, titre of anti-Campylobacterjejuni-IgG antibody in serum reach the peak at 2-4 weeks. From week 2, the serum A value was significantly higher in the experimental group than in the control group (1.923±0.403, 0.973±0.633, P ＜ 0.05). The IgG type GM1 (A value) was obviously elevated at week 8, but insignificantly different from that in the control group (0.115±0.042, 0.097±0.039, P ＞ 0.05). The MBP content (Avalue) in serum was significantly elevated at the 8th week (0.134±0.041).③ The imaging examination showed that abnormal MRI signal of different degree occurred at 2-4 weeks after immunization in the experimental group. ④ The histological changes showed that there was swelling of myelin sheath at the sites of brainstem, medulla oblongata, cervical spinal cord, thoracic spinal cord and lumbar spinal cord in the experimental group, no inflammatory cell infiltration and deletion of myelin sheath were observed. No obvious changes at the above site were observed in the contro1 group.CONCLUSION: Campylobacterjejuni Penner 4 can induce lesion of central nervous system.

ObjectiveTo observe the prepulse inhibition(PPI) ot the startle reflex of pure cerebral concussion (PCC) suffered from one concussion and multiple cerebral concussion (MCC) suffered from three concussions in rats,and to explore accumulate effect upon cognitive dysfunction of MCC.MethodsA metallic pendulum striker device for closed head injury was employed to duplicate PCC and MCC models in Stragu-Dawley rats.The MCC rats were hit three times on rats'head and it is interval 24 hours for every hit.According to the criteria of cerebral concussion,the investigated animals were divided into PCC group and MCC group at freedom.One control group was used.Each group included 10 animals.Each experience mental animal was tested from 3 days pre-injury to 28 days post-concussion.Startle reflex amplitude (for P values),pre-stimulation induced reflex amplitude on three standard stimulations,that was,67dB,69dB and 73dB (for PP67,PP69 and PP73 values) and prepulse inhibition of the startle reflex (PPI) were collected.ResultsThe P values and three PP values in the first three days of pre-replication experiment,there was no statistical significance in each group.However the P values and PP67,PP69 and PP73 values declined until to the 16th day after injury (P＜0.05),then recovered in PCC group.The P value and PP67,PP69 and PP73 values changes of MCC group declined and not recovered until to test end (P＜0.05 ) and they were more lower than PCC.The three PPI values were a little bit increase in both groups,there were statistics significance at some test points (P＜0.05) compared with control.ConclusionThe startle reflex amplitude and pre-stimulation induced reflex amplitude weaken after cerebral concussion and there is damaging accumulate effect to injury times,the PPI is enlanced by cerebral concussion.