Objective　To explore the role of tyrosine hydroxy lase (TH) gene in the treatment of Parkinson's disease (PD) by using ex vivo gene transfer. Methods　After the construction of TH gene in a retroviral vector, the astrocytes were cultured with the supernatant containing the recombinant DNA and then grafted into the cerebrum of PD rats. The reduction of the rat rotation beharior was evaluated. Results　The rotati on of PD rats was markedly improved in the rats with ex vivo gene transfer. Conclusion　TH has an obvious efficiency on the treatment of PD and the astrocytes can be used as effective gene transfer cells.

Objective To analyze the levels of Th17,CD4+ CDhigh25and the related cytokines (IL-17,IL-21,TGF-β,IL-10)in the plasma of patients with SLE.Methods The percentage of Th17 and CD4+ CDhigh25 Treg cells in peripheral blood were examined by flow cytometry.QRTPCR was used to detect the mRNA expression level of RORγt and Foxp3 of 56 SLE patients (26 patients are inactive and 30 patients are active) and 28 healthy controls.Plasma cytokines levels were measured by ELISA.Results The percentage of Th17 cells in patients with SLE was higher than that of healthy controls [ (3.44 ± 0.96) ％ vs (2.42 ± 0.52) ％,t =5.38,P =0.000 ].The percentage of CD4+ CDhigh25 Treg cells in patients with SLE was lower than that of healthy controls [ ( 1.32 ± 0.57 ) ％ vs ( 2.07 ± 0.67 ) ％,t =3.28,P =0.034].The expressions of RORγt mRNA was increased compared with healthy controls[ (0.219 ±0.063) vs (0.087 ± 0.045 ),t =6.41,P =0.000 ],but the expressions of Foxp3 mRNA was decreased compared with healthy controls [ (0.063 ±0.045) vs (0.128 ±0.056),t =5.28,P =0.000].Plasma levelsof IL-17,IL-21 and IL-10 in SLE patients were 122.4 ( 60.5 - 188.3 ),167.2 ( 128.7 - 871.4 ),51.3(20.9 - 123.7 ) ng/L,respectively,and in the control group were 27.1 ( 18.1 - 86.2 ),31.0 ( 31.0 -424.5),33.5( 16.4 - 54.1 ) ng/L,respectively,they were significantly different between the two groups (Z =3.83,4.54,1.87,all P ＜ 0.05 ).However,TGF-β levels were markedly decreased in SLE patients compared with controls [ 31.0 ( 31.0 - 168.6 ) ng/L vs 159.8 ( 63.4 - 389.7 ) ng/L,Z =4.87,P ＜0.05.IL-17 was positive correlation with Th17 cells,SLEDAI score,dsDNA antibody titers ( r =0.621,0.581,0.512,respectively,all P ＜ 0.05 ),but was negative correlation with C3 ( r =- 0.543,P ＜ 0.05 ) in SLE patients.Although the percentage of CD4 + CDhigh25 Treg cells were negatively correlated with SLEDAI score ( r =- 0.423,P ＜ 0.05 ),it was positively correlated with C3 ( r =0.511,P ＜ 0.05 ).Conclusion This study suggests that the imbalance of Th17 cells and CD4+ CDhigh25 Treg cells and the disorder of related cytokines are important in the pathogenesis of SLE patients.

Objective To analyze and evaluate the changes of quantity and function of TCBVα24+ Vβ11+ NKT in PBMC of the patients with PBC and its relationship with the occurrence of PBC. Methods Flow cytometry was utilized to count TCRVα24+ Vβ11+ NKT cells in PBMC in 60 cases of PBC and 60 cases of age-matched and gender-matched controls. NKT cells were activated and expanded by α-galcer and IL-2 in vitro. The percentages of positive NKT cells expressing IL-4 and IFN-γ were determined by ICS-FC. The levels of serum IL-4 and IFN-γ were tested by ELSIA. The numbers of cells secreting IFN-γ and IL-4 were detected by ELISpot. Results The ratios of NKT cells in PBC group were [0. 16(0. 11-0. 26) ]% before expansion and [0. 82 (0. 61-0. 89) ]% after expansion, significantly lower than control group [(0.33 (0.27-0.38) ]% and [27.40 (23.52-33.87) ]%, respectively, (Z=6.563, 7.707, P<0. 01 ). Seven days after expansion by α-galcer and IL-2, the expansion folds of NKT cells were 96. 05 (80.50-100.27) in PBC group and 134.65 (121.60-142. 13) in control group, respectively (Z =6.462,P < 0. 01 ). At the same time, the ratio of IFN-γ+ and TCRVα24+ Vβ11+ NKT detected by ICS-FC in PBC group was significantly higher than that in control group [ 38. 98 ( 36.73-42. 98 ) ]% vs [ 34. 56 ( 30. 12-36. 78 ) ] %, Z = 3. 158, P < 0. 05, while the ratio of IL-4+ and TCRVα24 + Vβ11+ NKT cells in PBC group was significantly lower than that in control group[0. 193(0. 179-0. 218) ]% vs [34. 36 (30. 93-38. 77) ]%,Z =6. 476, P <0. 01. The number of IFN-γ SFC detected by ELISpot were [410(380 ~500) ] SFC/1 × 106 PBMC in PBC group and [ 340(280 ~ 390)] SFC/1 × 106 PBMC in control group, respectively (Z = 4. 312, P <0. 05). The levels of serum IFN-γ in PBC group and control group were (67.21 ± 11.27) ng/L and (31.45 ± 8. 17) ng/L, respectively ( t = 27.25, P < 0. 01 ). The level of IFN-γ in PBC group was higher than that of control group. The number of IL-4 SFC were [73(60 ~ 100) ]SFC/1 × 106 PBMC in PBC group vs [245(230 -280) ] SFC/1 × 106 PBMC in control group, Z=5. 112,P <0. 01. The levels of serum IL-4 in PBC group and control group were (12.65 ±4. 17) μ/L and (28.31 ±6.31) μg/L, respectively (t =25.34,P < 0. 01 ). The level of IL-4 in PBC group was lower than that of control group. Conclusions The quantity of TCRVα2.4+ Vβ11+ NKT in PBC group is lower than that in control group. After in vitro activation, the capacity of expansion and producing IL-4 of NKT is decreased in PBC group, while the capacity of producing IFN-γ of NKT is increased in PBC group. The reduction of NKT cells and the immune dysfunction may be one of the important factors in the pathogenesis of PBC.

Objective To discuss the feasibility and safety of propofol used in senile patients undergoing minor surgery including gastroscopy and colonoscopy.Methods 40 patients aged 80-88 years old were randomly divided into two groups with 20 cases for each group.Group P received only propofol by intravenous injection in a speed of 1ml/s till eyelash response disappeared.In patients of group F fentanyl was first given at 0.5?g/kg,and 1 minute later,the patients were given propofol by intravenous injection in a same speed as in group P if there was no discomfort until eyelash response disappeared.MAP,HR,SpO2 and RR were determined before induction of anesthesia,beginning of operation,and 1st,3rd and 5th minute after operation.The onset time of anesthesia,total dosage of propofol used,time of palinesthesia and follow-up results were recorded.Results MAP was significantly decreased in both groups at the beginning of operation,and more stable in group F at the 3rd and 5th minute after operation.There were significant differences between two groups in amount of propofol used,and the dosage in group F was less than that used in group P(P

Objective To observe the effect of Cyclosporine A on the brain immunoligical reject reaction of transgenic cellular transplant in rats with experimental brain hemorrhage.Methods We established the brain hemorrhagic models of rats and the models were divided into group A (treated with Cyclosporin A) and group B (without treatment) randomly after transplanting brain cells modified by NGF gene. The levels of CD~+_4, CD~+_8T cells subsets in peripheral blood were examined by using flow cytometry at 15 d after transplantation. The expressions of MHC-classⅡantigens and infiltration of CD~+_4, CD~+_8T lymphocyte subsets were examined by SP immunocytochemical stain.Results The count of CD~+_4, CD~+_8T lymphocyte and CD~+_4/CD~+_8 in peripheral blood were 29.20?3.97, 20.65?2.02 and 1.41?0.86 in group A, and were 47.39?3.01, 28.30? 2.36 and 1.68?0.69 in group B, respectively.There were significant differences between the two groups (all P

Objective:To survey the status of anxiety/depression and lifestyle among junior high school students and explore their relation to put preference for psychological health education.Methods:Totally 1597 junior high school students aged 10 to 17 years in Shang Haicity were surveyed.The Generalized Anxiety Disorder-7 (GAD-7),Patient Health Questiormaire-9 items (PHQ-9) and Chinese Adolescent Lifestyle Scale (CALS),were used to assess anxious emotion,depressive emotion and lifestyle.Results:The rates of anxious emotion and depressive emotion were 16.4％ and 17.2％ respectively among students.Scores of GAD-7 and PHQ-9 were negatively correlated with all factor scores of CALS including eating habits,sleep status,health habits,learning habits,exercise,entertainment activities,interpersonal relationships,coping style and life satisfaction (-0.57 ≤ r ≤-0.15,Ps ＜ 0.01).Multivariate stepwise regression analysis showed that the sleep status factor scores,life satisfaction and exercise factor scores of CALS could negatively predict scores of GAD-7 (β3 =-0.17,-0.15,-0.05;Ps ＜ 0.05) and health habits positively predict scores of GAD-7 (β3 =0.08,P ＜0.001).Sleep status factor scores,life satisfaction factor scores and eating habits factor scores could negatively predict scores of PHQ-9 (β3 =-0.21,-0.16,-0.06;Ps ＜ 0.05).Conclusion:It suggests that the junior high school students'depression and anxiety are related to their lifestyle.The emotion of depression and anxiety of students may be predicted by some factors in the lifestyle.

Objective To discuss features of laparosco pi c cholecystectomy (LC) in children. Methods A total of 54 chi ldren underwent LC from October 1992 to December 2003. The operations were carri ed out after the creation of a CO2 pneumoperitoneum at the flow rate

BACKGROUND: A stable and exact animal model is the necessary tool and basis for studying hemorrhagic cerebrovascular diseases.OBJECTIVE: To establish and evaluate the intracerebral capsular hemorrhage models in rats.DESIGN: A randomized and controlled animal experiment.SETTING: Second Hospital Affiliated to Xuzhou Medical College; First Hospital Affiliated to Nanjing Medical University.MATERIALS: This experiment was carried out in the animal experimental center of Nanjing Medical University during May to November 2002.Totally 35 SD rats were randomized into two groups: experimental group (n=30) and sham-operation group (n=5).METHODS: ① Autoblood was injected into the intracerebral capsule of rats to create intracerebral capsule hemorrhage models with stereotaxy in the experiment group. ②Scoring was conducted according to 5-point neurological scoring criteria from ZeaLonga, somatic sensation and motor function of rats were observed. ③Somatosensory evoked potential(SEP) of rats was detected pre- and post-operation under anesthetic state. ④ After determination of SEP, the rats were sacrificed under anesthetic state. Brains were taken out to made slices, then sections were stained with haematoxylin and eosin. Changes in haematoma and histomorphology were observed at the largest focus under optical microscope.MAIN OUTCOME MEASURES: ①Nerve function scoring; ②Latent period of various waves of SEP; ③ Observation of brain tissue morphology.RESULTS: Totally 35 rats entered the stage of result analysis. ①Appearance of obvious paralysis of the rats suggested the modeling was successful. The successful rate of this experiment was 93.3%(28/30). Significant difference existed in neurological scoring between experimental group [(2.74±0.46)points] and sham-operation group (0 point)(P＜0.05). ②SEP showed that the latent periods of various waves of experimental group after operation were significantly delayed than those before operation and those of sham-operation group [P1: (15.72±0.78) ms, (10.69±0.52) ms, (10.73±0.48) ms;Nl: (17.95±1.27) ms,(13.21±1.31) ms, (13.34±1.27) ms;N2:(21.16±1.62) ms, (15.42±1.46) ms,(15.58±1.44) ms;N3:(24.86±1.58) ms, (18.72±1.76) ms, (18.99±1.67) ms,P＜0.05]. ③In the shamoperation group, a few red blood cells were scattered in the peripheral area of needle channel were found, but hemorrhagic focus was not; In the experimental group, irregular or oval blood clots presented in the left internal capsule area. In about a low-fold visual field, brain tissue in the surrounding of hemorrhagic focus was loosened and swelled, and pathological changes were obviously severer than those in the sham-operation group.CONCLUSION: Intracerebral capsular hemorrhage induced by injection of autoblood with stereotaxy is more close to clinical situation, and it is easy to operate and has good reproducibility.

Objective To Investigate the concentration of VILIP-1 and NR2 peptide in the serum of patients with ischemic stroke , and to explore their clinic value in early diagnostic of ischemic stroke patients.Methods The levels of VILIP-1 and NR2 peptide were examined by ELISA ( enzyme linked immunosorbent assay ,ELISA) with suspicious TIA ( defined as a neurological deficit that resolved within 24 hours) or acute ischemic stroke patients ( within 72 hours of onset of symptoms ) 340 cases,102 healthy controls,98 patients with vascular risk factors and 35 patients with hemorrhagic stroke.Among all the groups , VILIP-1 and NR2 peptide level were analyzed using the nonparametric Wilcoxon test.Diagnostic performance were analyzed among the groups with the two biomarkers independently and combinedly .Results Serum levels of VILIP-1 and NR2 peptide in patients with ischemic stroke (IS) were 9.80 (1.90-14.22) μg/L, 14.40 (5.60-27.91) μg/L respectively,which was higher than that of the healthy control group [VILIP-1:0.02 (0.01-0.09),NR2:0.33 (0.02-1.15),χ2 were 5.61 and 9.54,P<0.001],the group with vascular risk factors [VILIP-1:0.03 (0.02-0.16),NR2:0.27 (0.01-1.54),χ2 were 6.74 and 10.62,P<0.001], the group of patients non-stoke [VILIP-1:0.04 (0.03-0.19),NR2:0.53 (0.45-1.21),χ2 were 3.78 and 7.63, P <0.001 ].The levels of VILIP-1 and NR2 peptide was significantly increased in IS patients presenting within 3 h of symptom onset.When differentiating IS from patients with hemorrhagic stroke ,NR2 had a AUC of 0.934,showing a strong distinguishing effectiveness.Differentiating IS from healthy controls , patients with vascular risk factors and non-stroke patients,the AUC of combination of VILIP-1 and NR2 was 0.974,which was higher than the AUC of either VILIP-1(0.849) or NR2(0.862) alone(P <0.05). Conclusions VILIP-1 and NR2 peptide are very sensitive and specific biomarkers to the early diagnosis of IS.The combination of VILIP-1 and NR2 peptide has higher value of clinical applications than one of them independently.

Objective To determine the icariin content in Qianlie Shule Granules (QSG). Methods After proper pre-treatment,HPLC method was performed on a C18 column. The mobile phase consisted of acetonitrile-water (29 ∶71) and the UV d etective wavelength was set at 270 nm. Results The peak of icariin was separated completely. The calibration curve of icariin was linear within 0.16～1.2 mg?mL -1,Y=0.028 7X-0.009 62 (r=0.999 9). Conclusion This method is accurate and su itable for the determination of icariin content.