Objective To explore the therapeutic effects of human glial cell line-derived neurotrophic factor (GDNF)-engineered rat neural stem cell (NSC) transplantation in rat model of Parkinson's disease ( PD) . Methods SD rats received a single injection of 24 μg of 6-hydroxydopamine (6-OHDA) at two sites in right striatum. Then 10 days after surgery, the successful animal models of PD were divided into 3 groups: PD model group ( 2 μl transplantation media was injected in right striatum), NSC group (transplanted were 2×10~5 NSCs infected by bare lentivirus) and GDNF group (transplanted were 2×10~5 GDNF-engineered NSCs). The rotation scores were assessed 5 weeks, 7 weeks and 9 weeks after transplantation. The dopaminergic neurons in substantia nigra ( SN ) were analyzed quantitatively using immunohistochemistry for tyrosine hydroxylase (TH), and the dopamine and its metabolites, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were analyzed 9 weeks after transplantation by high performance liquid chromatography ( HPLC) . Results GDNF-engineered NSC transplantation could effectively improve the behavioral performance in rats. At the 5th week after cell transplantation, the rotation turns within 90 min were (993. 9±159. 1) turns, (956. 7±136. 3) turns and (433. 6±100. 9) turns in PD model group, NSC group and GDNF group respectively (F=95. 694, P = 0. 000). At the 7th week, the rotation turns within 90 min were (964. 2 ± 152.0) turns, (909. 2 ± 136. 3) turns and (399. 4±84. 4) turns in PD model group, NSC group and GDNF group respectively (F = 106. 134, P=0. 000). At the 9th week, the rotation turns within 90 min were (909. 5±152. 2) turns, (865. 5± 129. 1) turns and (312. 2±63. 7) turns in PD model group, NSC group and GDNF group respectively (F= 151. 100, P = 0.000). GDNF-engineered NSC transplantation could significantly increase the levels of dopamine and its metabolites in injured striatum. The concentrations of dopamine in injured striatum was higher in GDNF group than that in PD model group and NSC group C(7. 5±0. 8) ng/mg vs. (3.3±0.3) ng/mg and (3. 7±1. 3) ng/mg, F=59. 543, P = 0. 0003. The level of DOPAC was higher in GDNF group than that in PD model group and NSC group C(0. 9±0. 1) ng/mg vs. (0. 5± 0. 1) ng/mg and (0. 6±0. 2) ng/mg, F= 17. 293, P=0. 000]. The concentration of HVA in injured striatum was higher in GDNF group than that in PD model group and NSC group [(0. 9±0. 1) ng/mg vs. (0.5±0. 1) ng/mg and (0. 6±0. 2) ng/mg, F=35.175, P = 0.000]. Conclusions engineered NSC transplantation improves the function of dopamine system in SN and striatum, and GDNF gene therapy has potential clinical value.

Objective To identify the cell death type and investigate the potential mechanism of ionizing radiation-induced neural cell death in the neonatal rat cerebral cortex.Methods The neonatal Wistar rats were given a single dose of 2.0 Gy γ-irradiation.The cell death type and characterization in cerebral cortex were identified using DNA electrophoresis,TUNEL and HE staining.The P53-and iNOS-positive cells were analyzed quantitatively using immunohistochemistry.Results The DNA and morphological characterization of death cells indicated that 2.0 Gy γ-irradiation induced apoptosis of the neural cells in neonatal rat cerebral cortex.The apoptosis indices in different cortex regions were significantly increased 4 h after irradiation,and reached the peak value at 12 h post-irradiation.The apoptosis index of neoconex was much higher than that of hippocampus(archicortex)and paleocortex,while paleocortex had lower apoptosis index than hippocampus.The quantitative immunohistoehemistry suggested that the numbers of P53 and iNOS-positive cells were not different between these three cortex regions at the same time-point after irradiation.Conclusion 2.0 Gy γ-rays induced apoptosis of the neural cells in neonatal rat cerebral codex.The response of cells to the damage effects of ionizing radiation was similar in different cortex regions;however,the apoptosis indices were different significantly.These findings imply that the developing phase or type of neural cells may play a pivotal role in the apoptosis process induced by ionizing radiation.

Objective To investigate the efficacy of milrinone micropump in treatment of chronic heart failure(CHF),and the influence to the level of amino-terminal B-type natriuretic peptide precursor(NT-proBNP) in serum.Methods 120 cases with CHF were selected,they were randomly divided into the observation group and the control group according to a random number table.The patients of the control group were treated with standardized anti -conventional treatment of heart failure according to their heart function,the patients of the observation group were given milrinone micropump on the basis of the standardized anti-conventional treatment of heart failure.The efficacy of clinical conditions 1 week after treatment and NT-proBNP levels in serum before and after treatment were compared between two groups.Results The clinical efficacy of the observation group was 95.00％,which was higher than 76.67％ of the control group,the difference was statistically significant(x2 =8.67,P ＜ 0.05).The serum level of NT-proBNP of the observation group was (38.9 ± 9.2) ng/L,which was significantly lower than (86.3 ± 32.4) ng/L of the control group,the difference was statistically significant (t =9.86,P ＜ 0.05).Conclusion Milrinone micro pump used in the treatment of CHF can significantly improve the clinical efficacy,and lower the serum level of NT-proBNP,improve heart function,it is safe and reliable and worth to application in the clinical.

Objective To discuss the clinical application of MSCT-guided BARD gun biopsy in diagnosing pulmonary lesions. Methods Under MSCT-guidance percutaneous transthoracic biopsy of lung with BARD gun was performed in 68 patients with pulmonary lesions. Results All the procedures were well performed with the technical successful rate of 100%. The final diagnosis was based on the pathological findings or clinical follow-up observation. The accuracy of the qualitative diagnosis was 94.1%. Of 68 patients, squamous cell carcinoma was found in 30, adenocarcinoma in 21, metastases in 7, inflammatory pseudotumor in 3, lung abscess in 2, and tuberculosis in one. The false-negative rate in making diagnosis was 5.9%. The complication occurrence was 8.8%. Conclusion BARD gun has the advantage of making coaxial- multipoint biopsy. With the help of MSCT-guidance and the full use of the operating advantage of BARD gun, this technique is safe and reliable, with higher accuracy and less complications.

Objectives This study was to describe the prevalence of depressive and/or anxiety symptom and disorder in outpatients with irritable bowel syndrome (IBS) or functional dyspepsia (FD) in the comprehensive hospitals in big cities, China. Methods This was a hospital-based cross-sectional study conducted in three main cities in China in 2004. Six hundred and eight eligible subjects with IBS or FD were recruited consecutively from the outpatient departments within 3 months. Face-to-face interview was used in data collection together with the self-completed HAD scale for depressive and/or anxiety symptom screening. Subjects getting a HAD score of 9 and above were further assessed for depressive and/or anxiety disorders with HAMA scales and HAMD scales by the licensed psychologists or psychiatrists. Results The prevalence of depressive and/or anxiety symptoms was 24. 8% and 23. 6% respectively in outpatients with IBS or FD. And among detected depressive and/or anxiety symptoms, the prevalence of depressive and/or anxiety disorder was 69. 6% and 68. 7% respectively; Less than 12% of subjects had obtained a diagnosis of depressive disorders and been treated, and less than 8% of subjects had obtained a diagnosis of anxiety disorders and been treated prior to the investigation. The prevalence of anxiety symptom in female patients with IBS was significantly higher than male patients (17. 0% vs. 8. 7%; X2=4. 432,P = 0. 035); among detected depressive and/or anxiety symptoms, the prevalence of depressive disorder, anxiety disorder, depressive and anxiety disorder and depressive and/or anxiety dis order in female patients with IBS were higher than those in male patients (52. 6%vs. 25. 0% ,X2 = 4. 612, P=0. 032; 79. 5% vs. 53. 8%,X2=4. 815, P = 0. 028; 51. 4% vs. 21. 7%,X2=5. 112,P = 0. 024; 78. 6% vs. 55. 6%,X2=4. 112,P = 0. 043). Conclusions It is important for healthcare providers and health policy makers to note that the prevalence of depressive and/or anxiety symptoms and depressive and/or anxiety disorders are high and the percentages of previous diagnosis and treatment in outpatients with somatic diseases in general hospitals of urban China are rather low.

Objective To investigate dynamic change of anterior pituitary hormones (APHs), thyroid func-tion (TF) and genital hormones (GnHs) in patients with traumatic brain injury (TBI) and their clinical signifi-cance. Method APHs, TF and GnH were tested in 93 patients with TBI,who were admitted to Zhejiang Provin-cial People's Hopital from March 2006 to June 2007. Patients with primary injury in the hypothalamic and pituitary regions, as detected by CT and/or MR/examination, as well as those with tumors or immune diseases in the CNS,endocrine or urinogenital systems, were excluded. The clinical data were analyzed according to Glasgow coma scores (GCS), type and degree of injury, and whether there was any secondary cerebral injury. Twenty healthy people acted as controls. The data were analyzed by the Hotelling T2 test and t-tests using SAS 11.5. A P value of less than 0.05 indicated statistical significance. Results The levels of adrenocortieotropic hormone (ACTH),luteinizing hormone (LH) and prolactin (PRL) were markedly higher in all 93 TBI patients than controls, while those of thyroid-stimulating hormone (TSH), thyroid hormone T3,T4 and FT3 were significantly lower in TBI pa-tients in the early stage after injury than in those at follow-up and controls (P<0.05). The ACTH and PRL val-ues reached (33.33±6.86) and (31.74±5.51), respectively, and the LH value was (9.48±1.14) in the secondary cerebral injury group.The TSH value (1.26±0.17) in the brain injury group was significantly lower than those in controls (P<0.05). With the exception of TSH, PRL, testosterone (T) and E2, other APHs were markedly lower in TBI patients at following-up than in controls (P<0.05). The incidence of traumatic hypothala-mus-pituitary insufficiency (THPI) associated with low levels of more than three APHs was 3.2%, while 13.8% of THPI patients showed low levels of at least one APH. The ratio of sick euthyroid syndrome (SETS) was 14.0%. Conclusions A low level of a single APH is the prevalent pattern in THPI patients. Secondary cerebral injury, such as acute high intracranial pressure, brain edema and ischemia after TBI, may be the chief causes of THPI. Early hyperprolactinemia is an important indication for presaging THPI. The dynamic levels of neurcen-docrine hormones can serve as an important index for determining the suitability of TBI patients for treatment with hormone therapy.

Objective To evaluate the value of MSCTA in combination with CTV in diagnosis of pulmonary embolism and deep venous thrombosis (DVT) of lower limb. Methods 47 patients with pulmonary embolism proved clinically were examined with MSCTA using 120 kV,210 mA,collimater of 0.75 mm and the automatic bolus traching technique,the speed of contrast agent infu-sion was 3～4 m/s after 170 s delayed,venous scan of lower limbs was performed. Results 47 cases were well perform the trunk of bilateral pulmonary artery, pulmonary lobar artery, pulmonary segemental artery, pulmonary inferior segmental artery could be showed by MSCTA, MPR, MIP and VRT. 363 arterial segments were involved and 31 cases accompanied with DVT. Conclusion MSCTA in combination with CTV is of significant value in detecting pulmonary embolism accompanied with deep venous thrombo-sis.

Objective To explore the mechanism through which nicotine protects dopaminergic neurons against 6-OHDA toxicity in SD rat. Methods Rats received nicotine or saline treatment (two doses tested,0. 2 rag/ kg and 2 rag/ kg, 5 injections i.p. per day at 2-h intervals). On day 8after the treatment, a single injection of 20μg of 6-OHDA was administered into right striatum.Nicotine or saline was administered continuously daily until animals were killed. The dopaminergic neurons and CD3, CD4 and CDS-positive lymphocytes were analyzed quantitatively using immunohistochemistry. Microglia activation was quantified by IBA1 immunofluorescence. Results The loss of dopaminergic neurons induced by 6-OHDA in the substantia nigra was significantly less severe in the nicotine treatment group (at both 0. 2 and 2 mg/kg groups) than that in the saline treated group. In the striatum, we observed that the number of CD3, CD4 and CD8-positive lymphocytes reduced significantly in the nicotine treated animals as compared to saline controls. Otherwise, nicotine inhibited CD4 and CD8-positive lymphocytes infiltration equivalently. Quantitative immunofluorescenee analysis indicated the microglia activation was inhibited obviously in nicotine treatment. Conclusions Our data suggest that nicotine may have a neuroprotective effect against dopaminergic lesion induced by 6-OHDA by inhibiting the inflammation.

BACKGROUND:Whether transplanted bone marrow mesenchymal stem cells under hypoxic conditions can survive is crucial for the successful celltransplantation. Therefore, studies on the growth of bone marrow mesenchymal stem cells under hypoxic conditions in vitro can provide experimental evidence for in vivo celltransplantation. OBJECTIVE:To observe the expression of vascular endothelial growth factor in bone marrow mesenchymal stem cells under hypoxia. METHODS:Rat bone marrow mesenchymal stem cells were obtained and cultured, and observed under light microscopy. Passage 3 cells were cultured under normoxia (21%O2) and hypoxia (3%O2 hours. Then cellcounting kit-8 assay and flow cytometry were employed to detect cellproliferation in the two groups. Western blot assay was adopted to detect the expression of hypoxia-inducible factor-1αand vascular endothelial growth factor in the two groups. RESULTS AND CONCLUSION:(1)Rat bone marrow mesenchymal stem cells were obtained and cultured successful y, which were fusiform cells and had uniform shape under the light microscope. (2)The results of cellcounting kit-8 assay showed that the number of cells in the hypoxic group was higher than that in the normoxic group at each time point, and cellviability increased significantly at hours 36 and 48 (P<0.05). (3)The results of flow cytometry demonstrated that the proportion of cells in S phase and cellproliferation index in the hypoxic group were significantly increased, compared with the normoxic group (P<0.05). (4)Western blot results showed ), respectively, for 72 that there was a smal amount of the expression of hypoxia-inducible factor-1αand vascular endothelial growth factor in the normoxic group, but the expression of these two proteins in the hypoxic group was increased in a time-dependent manner (P<0.05). These findings suggest that hypoxia can induce proliferation of rat bone marrow mesenchymal stem cells cultured in vitro, and also raise hypoxia-inducible factor-1αand vascular endothelial growth factor expression in a time-dependent manner.

BACKGROUND:Caspase plays a crucial role in the cellapoptosis, but the influence of different facial nerve injury on caspase 1, caspase 8, cyto-c protein expression and their correlation stil remain unclear.
OBJECTIVE:To construct facial nerve crush or distal transection injury models, observe the morphological changes of facial motoneurons, investigate death gene caspase 3, caspase 8, cyto-c expression, and analyze their correlation.
METHODS:Facial nerve crush or distal transection injury model was established in the right facial nerve of rats, while the left facial nerve served as normal controls. We observed the morphology and the death of facial motoneurons with toluidine blue staining and transmission electron microscope. Expressions of caspase 3, caspase 8 and cyto-c proteins were studied by immunohistochemistry analysis fol owing facial nerve injury.
RESULTS AND CONCLUSION:Both facial nerve distal transection and crush injury resulted in the death of facial motoneurons, and the death pattern was mainly apoptosis. Caspase 3, caspase 8 and cyto-c protein expressions were observed in the subnucleus of normal rat facial nucleus. cells of the distal transection group were stained more intensely than that of crush group. Expressions of these proteins began to increase at 3 days after the injuries. Caspase 3 and caspase 8 protein expression peaked at 14 days, whereas cyto-c protein expression peaked at 7 days after the injuries. Expressions of caspase 3, caspase 8 and cyto-c proteins were correlated with facial nerve injury type and injury time. Expressions of caspase 8 and cyto-c protein were correlated with expression of caspase 3 protein. The findings indicate that, caspase 8 and cyto-c contribute to activate caspase 3, and caspase cascade reaction plays an important role in the apoptosis of facial motoneurons.