OBJECTIVE:To observe the distribution of fluorescent material in the respiratory tract of animals with the use of small animal imaging technology to quickly evaluate the effects of respiratory drug delivery. METHODS: The Kunming mice in the experimental group were administered with Cy5.5 fluorescent-labeled aerosol,meanwhile,a positive control group was set up by injecting same amount of compound anhydrous diethyl ether solution via tracheal incubation,with negative control group inhaled the same amount of non-Cy5.5 fluorescently-labeled aerosol. The fluorescent distribution of the aerosolized particles in the respiratory tract was observed. RESULTS: Fluorescently-labeled aerosol was observed in both the experimental group and the positive control group but not in negative control group. CONCLUSION: The small animal imaging technology is expected to be used as a rapid and effective method for evaluating the efficacy of respiratory drug delivery.

ObjectiveTo determine the effect of unfractionated heparin (UFH) on lipopolysaccharide (LPS)-induced expression of granulocyte colony-stimulating factor (G-CSF), and the role of Toll-like receptor 4 (TLR4) signaling pathway in this process.Methods Human pulmonary microvascular endothelial cells (HPMECs) were cultured in vitro, and the cells between passages 3 and 5 were used in the experiments. ExperimentⅠ: the cells were divided into four groups as follows: control group, LPS stimulation group (LPS 10μg/mL), LPS+ 0.1 U/mL UFH group, and LPS+ 1 U/mL UFH group. HPMECs in UFH groups were treated with 0.1 U/mL or 1 U/mL UFH 15 minutes before LPS stimulation, and HPMECs in control group were treated with an equal volume of phosphate-buffered saline (PBS) instead. The concentrations of interleukin-6 (IL-6) and G-CSF in cell culture supernatants were determined by enzyme linked immunosorbent assay (ELISA) 24 hours after LPS challenge to detect the effect of UFH on HPMECs. ExperimentⅡ: HPMECs were treated with 5μg/mL of rhodobacter sphaeroides LPS (LPS-RS, antagonist for TLR4) 4 hours before the addition of PBS or LPS. The concentrations of IL-6 and G-CSF in cell culture supernatants were determined 24 hours after LPS stimulation to detect the effect of TLR4 on LPS-induced HPMEC injury. ExperimentⅢ: HPMECs were divided into four groups as before: control group, LPS stimulation group, LPS+ 0.1 U/mL UFH group, LPS+ 1 U/mL UFH group. Treatments to cells were the same as experimentⅠ. The protein expression of TLR4 in HPMECs was determined by Western Blot 1 hour after LPS stimulation to detect the effect of UFH on TLR4.Results① Compared with control group, the levels of IL-6 and G-CSF in LPS stimulation group were increased [IL-6 (ng/L): 655.9±58.3 vs. 75.5±18.2, G-CSF (ng/L): 388.7±36.2 vs. 35.3±12.6, both P< 0.05]. Compared with those of LPS stimulation group, in LPS+ 0.1 U/mL UFH group and LPS+ 1 U/mL UFH group, the levels of IL-6 and G-CSF were significantly decreased [IL-6 (ng/L): 518.2±64.6, 489.1±75.6 vs. 655.9±58.3, G-CSF (ng/L): 298.8±41.0, 273.4±33.2 vs. 388.7±36.2, allP< 0.05]. The results indicated that 1 U/mL UFH had better results, though there was no statistical significance between the results of two UFH groups.② LPS-induced up-regulation of IL-6 and G-CSF levels was prevented by LPS-RS [IL-6 (ng/L): 139.1±37.6 vs. 655.9±58.3, G-CSF (ng/L): 73.7±19.7 vs. 388.7±36.2, bothP< 0.05]. LPS-RS alone had no effect on cytokines [IL-6 (ng/L):118.2±42.1 vs. 75.5±18.2, G-CSF (ng/L): 48.4±26.8 vs. 35.3±12.6, bothP> 0.05].③ Compared with control group, the protein expression of TLR4 (grey value) in LPS stimulation group was significantly upregulated after 1 hour (0.87±0.23 vs. 0.36±0.12,P< 0.05). UFH with 0.1 U/mL and 1 U/mL lowered TLR-4 protein expression induced by LPS (0.68±0.18, 0.62±0.26 vs. 0.87±0.23, bothP< 0.05).ConclusionsThe expressions of IL-6 and G-CSF were increased obviously in LPS treated HPMECs. UFH might take its therapeutic effect through TLR4-dependent pathway.

Objective To investigate the analgesic effect of auto-moxibostion on lumbar vertebral osteoarthritis? Methods Sixty patients with lumbar vertebral osteoarthritis were randomized equally into the control group and observation group by a random digits table: the former group received routine nursing care and the latter was intervened with auto-moxibustion together with routine nursing care? The curative effects were appraised by short-form of McGill pain questionnaire? Result The curative effect in the observation group was significantly better than that of the control group(P < 0?05)? Conclusions The auto-moxibustion is effective for the analgesia of lumbar vertebral osteoarthritis? The right management of medication and detained medicine is helpful for the increased effect of auto-moxibustion?

AIM: To observe the effect of phosphorylation protein kinase C delta (PKC?) on the procedure of PC12 cells apoptosis induced by 6-hydroxydopamine(6-OHDA) and to investigate the potential molecular pathogenesis of Parkinson disease.METHODS: TUNEL staining and transmission electron microscope were applied to measure apoptosis when dopaminergic PC12 cells exposed to the excitomotors and inhibitors of PKC before 6-OHDA for 18 hours. The expression of phosphorylation of PKC? was detected by Western blotting. RESULTS: PMA, an activating agent of PKC?, significantly increased PC12 cell apoptosis induced by 6-OHDA. Rottlerin, an inhibitor of PKC?, protected PC12 cells apparently. As contrast, bisindolylmaleimide I, an inhibitor of general PKC and G6976, the inhibitor of calcium-dependent PKC, did not show any protective role. CONCLUSION: The phosphorylation PKC? is one of the important links in the process of PC12 cell apoptosis induced by 6-OHDA. PKC? may directly participate in neurodegeneration process in parkinsonian.

AIM: To observe the role of peroxysome proliferator activated receptor-?(PPAR-?) and the relationship of cyclooxygenase-2(COX-2) and PPAR-? in injury of cultured rat cortical neurons induced by hypoxia/reoxygenation.METHODS: Primary rat cortical neurons were cultured.Experiments include control group,hypoxia/ reoxygenation group and hypoxia/ reoxygenation with PPAR-? agonist group.Cell viability was surveyed by MTT assay.COX-2 protein expression was measured by Western blotting.RESULTS: Neuron viability raised dramatically in hypoxia/reoxygenation with PPAR-? agonist group,compared with hypoxia/reoxygenation group(P

BACKGROUND: Evidence exists that inhibition of matrix metanoproteinase-2(MMP-2) secretion in the proliferating hernangioma tissue by transfection of adenovirus-active MMP-2(Ad-aMMP-2) cDNA would become an important means for treatment of proliferating hemangioma.OBJECTIVE: To investigate the influences of Ad-aMMP-2 cDNA transfection on human proliferating hemangioma growth in nude mice.DESIGN, TIME AND SETTING: A randomized, grouping, and controlled observation was performed in West China Hospital of Sichuan University between August 2003 and September 2004.MATERIALS: Eighteen BALB/c-nu/nu nude mice, weighing approximately 20 g, were included. Cavernous hemangioma specimen pathologically confirmed as proliferating hemangioma was resected from one 52-day-old female child patient.METHODS: The freshly reseoted human proliferating hemangioma specimen was sliced into small pieces with a size of 5 mm×4 mm×3 mm and subcutaneously implanted into the back of 18 nude mice within 1 hour to develop mouse models of hemangioma.Forty-five days after hemangioma implantation, 15 successful hemangioma nude mice were treated by intratumoral administration of adenovirus green fluorescent protein (Ad-GFP1 n = 51 Ad-GFP group), adenovirus-active MMP-2 (n = 5, Ad-aMMP-2 group), or the same amount of phosphate buffered saline (PBS1 n = 51 control group). Intratumoral administration was performed once every other day, for a total of 4 times.MAIN OUTCOME MEASURES: Observation of tumor volume and compadson of tumor necrosis area among 3 groups; detection of GFP expression in nude mouse; gross, hematoxylin-eosin staining, and transmission etectron microscope observation of tumor tissue morphology; determination of MMP-2 cDNA expression and microvascular density by immunohistochemistry; and detection of growth cycle and apoptosis of tumor cells by flow cytometry.RESULTS:①Ad-aMMP-2 could inhibit hemangioma growth in vivo, without marked adverse reactions. Tumor necrosis of different degrees was found in each group, and tumor necrosis area was significantly greater in the Ad-aMMP-2 group than in the control and Ad-GFP groups (P < 0.01). ②Histological sections displayed GFP gene expression in the Ad-GFP group. ③Gross observation results revealed relatively large tumor tissue in the control and Ad-GFP groups and relatively small tumor tissue in the Ad-aMMP-2 group. Hernatoxylin-eosin staining results showed that in the control and Ad-GFP groups, endothelial cells aggregated together in strip-shaped or lump-shaped appearance, and in the Ad-aMMP-2 group, there were many necrotic loci arranging in lamellar-shape appearance. Transmission electron microscope results revealed vascular endothelial cells with normal morphology in the control group and tumor cells with apparent nucleoli in the Ad-GFP group, while in the Ad-aMMP-2 group, some vascular endothelial cells exhibited chromatin pycnosis in the nucleus, forming apoptotic bodies.④ MMP-2 expression and microvascular density were significantly reduced in the Ad-aMMP-2 group than in the Ad-GFP and control groups (P < 0.05). ⑤The percentage of tumor cells in G0/G1 phase was significantly higher (P < 0.05), while the proliferating index was significantly decreased, in the Ad-aMMP-2 group than in the Ad-GFP and control groups. The Ad-aMMP-2 group exhibited higher apoptosis rate of tumor cells (P < 0.05), as well as more markedly increasing apoptosis index, than the control and Ad-GFP groups.CONCLUSION: It is feasible to block human proliferating hemangioma growth by transfeotion of Ad-aMMP-2 cDNA. The included mechanisms are to inhibit vascular endothelial cells to secrete MMP-21 thereby leading to local ischemia.

Objective To detect the expression of scleroderma-related autoantibodies, such as anti-Scl-70, anli-centromere antibody ( ACA)and anti-RNA polymerase Ⅲ ( ARA) , and their relationship with clinical features in Chinese systemic sclerosis (SSc) patients. Methods One hundred and thirty-five Chinese SSc patients from the clinical database of the Scleroderma Trials and Research Group proposed by European League Against Rheumatism's Scheroderma Trial and Research Group( EUSTAR) were consecutively enrolled. The expression of ARA, anti-Scl-70 and ACA were detected through linear immunoblotting, double immunodiffusion and indirect irnmunofluorescence, respectively. The relevance between the existing of autoantibodies and clinical manifestations was analyzed statistically. Results Among the 135 Chinese SSc patients, the prevalence of anti-Scl-70, ACA, ARA were 49. 6% , 13.3 % and 8.9% respectively. Patients with anti-Scl-70 antibody had significantly shorter disease course [(71 ±59) month vs (90 ± 103) month, P = 0.041] , higher proportion of interstitial lung disease ( P = 0. 031) but lower of pulmonary arterial hypertension (P =0.042). Modified Rodnan's skin score (P=0.008) and prevalence of facial and cervical cutaneous sclerosis (P = 0. 002) , distal (to elbow/knee ) cutaneous sclerosis ( P = 0. 004 ) and digital pitting scarring/disappear of digital pad were all significantly higher in anti-Scl-70 positive group. Patients with AC A had longer disease course ( P = 0. 036) , lower IgM level ( P = 0. 045) and were less prevalent of interstitial lung disease ( P =0. 045). Patients with ARA had higher serum creatinine and urea nitrogen level ( P ＜ 0.001) although otherwise features had unremarkable differences. Conclusion Scleroderma-related autoantibodies have relevance with different clinical manifestation and detection of these autoantibodies may be helpful to the diagnosis of SSc, organ involvement evaluation and predicting outcomes. The clinical relevances of autoantibodies in Chinese SSc patients may differ from other areas or races.

Objective To investigate the clinical and laboratory characteristics of SSc patients in China. Method The data of 119 consecutive SSc patients based on EUSTAR DATABASE in Peking Union Medical College Hospital from February 2009 to January 2010 were prospectively collected and analyzed. All patients fulfilled ACR classification criteria in 1980 for SSc. Thex2 test and t-test were used to analyze the data. Results (1) Demographic data. Sex ratio (F/M) was 109/10 and the age rang was (44±12) years. There were 65 diffuse cutaneous SSc (dcSSc) patients and 54 limited cutaneous SSc (1cSSc) patients. 112 patients (94.1%) had Raynaud's phenomenon (RP), and the age of RP occurrence was 36 years (13～76 years), among which it was the initial presentation in 91 patients (81.3%) and the disease duration from RP to other manifestation was 12 months.(2) Clinical manifestations. ① The gastrointestinal manifestations (70.6%), especially esophageal involvement (56.3%), articular involvement (54.6%), pulmonary interstitial fibrosis (PIF) (58.8%) were frequently observed, but renal crisis (2.5%), heart block (0) and reduced LVEF (0) were rarely detected. ② Twenty cases (28.6%) out of 70 PIF patients denied any respiratory symptom and were confirmed by HRCT screening. The disease duration from RP to PIF was 34 months(3～352months); 3 case of 24 pulmonary artery hypertension (PAH) patients had no clinical manifestations. The disease duration from RP to PAH was 25 months (4～343 months). Nineteen patients had PIF and PAH simultaneously. ③Peripheral artery disease: SSc patients had a lower ankle brachial index (ABI) level (0.91± 0.19 vs 1.09±0.08, P<0.00l). (3) Laboratory finding. All patients had positive ANA. The positive rate of anti-Scl-70 antibody and ACA was 56.0% and 14.7% respectively. There was no serum sample positive for anti-Scl-70 antibody and ACA. The positive rate of anti-RNA polymerase Ⅲ antibody was 13%. (4) Compared the clinical characteristics and laboratory findings between dcSSc and lcSSc patients, we found that there were significant differences between dcSSc and lcSSc patients in finger ulcer (40.0% vs 20.4%), ACA positive rates (7.7% vs 23.3% , P<0.05). Conclusion The system involvements besides skin in SSc is common, especially PIF and gastrointestinal involvement. According to our data, there are fewer cases with renal crisis and heart block. Because part of patients with PIF have almost no clinical manifestations, so early screening for PIF/PAH is important for early diagnosis and intervention.

BACKGROUND: The development of safe and powerful antidepressant agents from traditional Chinese herbs has become a hotspot in studies on anti-depression therapy. OBJECTIVE: To investigate the anti-depressive effect and possible mechanism of curcumin by behavioral and neurochemical procedures. DESIGN: Randomized grouping design and controlled experiment. SETTING: Depart, ment of Pharmacology, School of Basic Medical Sciences of Peking University.MATERIALS: This study was carried out in the laboratory of the Department of Pharmacology, School of Basic Medical Sciences of Peking University, between November 2003 and October 2004. A total of 240 male ICR mice were recruited. METHODS: The whole experiment was divided into 4 tests. ① Antagonism of reserpine-induced hypothermia: Totally 60 mice were randomly chosen and divided into 6 groups: normal control group, groups of various doses of curcumin (1.25, 2.50, 5.00 and 10.00 mg/kg), and positive control group (imipramine 10 mg/kg). Normal temperature of the mice was measured before experiment. The animals were given a single injection of reserpine (2.5 mg/kg). The mice were administered with drugs 18 hours later, namely, curcumin of different concentrations by gastric perfusion, groundnut oil (0.1 mL/10g by gastric perfusion) as well as imipramine (10 mg/kg by intraperitoneal injection). Rectal temperature was measured 60, 90, 120,150 and 180 minutes after administration, respectively. ② Potentiation of 5-hydroxytryptophan (5-HTP)-induced head twitches: animal grouping was the same as above, and the drug in positive control group was replaced by fluoxetine. The mice received gastric perfusion and the dose of curcumin given was the same as above. Groundnut oil and fluoxetine (10 mg/kg) and 5-HTP (70 mg/kg) were injected into the vein of the tail one hour later.The number of head twitches was counted within 5-10 minutes after 5-HTP treatment. ③ Antagonism of apomorphine-induced hypothermia: Mice grouping was the same as above; the drug in positive control group was replaced by imipramine. Curcumin was give as above at 4 doses, and groundnut oil and imipramine were also given. Large-dose apomorphine was injected subcutaneously (16 mg/kg). Rectal temperature was measured before injection, as well as 30 minutes and 60 minutes after injection. ④Determination of monoamine and metabolites: Mice grouping was the same as above. The drug in positive control group was replaced by imipramine.Curcumin was give as above at 4 doses, and groundnut oil and imipramine were also given. The content of monoamine and metabolites in the mice was measured with high performance liquid chromatography. ⑤ Dunnett's t test was used for comparison between groups.MAIN OUTCOME MEASURES: ① In reserpine-induced hypothermia test, the change of body temperature before and after administration. ② In 5-HTP-induced head twitches test, whether the times of head twitches were increased. ③ In apomorphine-induced hypothermia test, the change of body temperature after administration. ④ Effect of drugs on the content of monoamine.RESULTS: Totally 240 mice entered the result analysis. ① Experiment results of reserpine-induced hypothermia: Curcumin (5 mg/kg and 10 mg/kg)produced an antagonism against reserpine-induced hypothermia, and the results were significantly different from those in control group (P ＜ 0.05,P＜0.01). Curcumin of 10.00 mg/kg produced the similar effect compared as that of imipramine in positive control group. ② Results of 5-HTPinduced head twitches: Curcumin (5 and 10 mg/kg) could significantly increase the times of 5-HTP-induced head twitches (P ＜0.05, P＜0.01). ③Results of apomorphine-induced hypothermia test: 2.50, 5.00 mg/kg and 10.00 mg/kg of curcumin could significantly increase the content of 5-HTP, and 10 mg/kg of curcumin could significantly increase the content of norepinephrine and dopamine. There was significant difference from that in control group (P ＜ 0.05). By contrast, curcumin had no obvious effect on the content of metabolite 5-hydroxyindol acetic acid and 3,4-dihydroxyphenylacetic acid. Imipramine of 10 mg/kg as the positive control drug could significantly increase the content of 5-hydroxyindol acetic acid and norepinephrine (P＜0.05).CONCLUSION: Curcumin has an antidepressant effect and the effect exerted may be related to monoaminergic neurotransmitter system.

Objective To observe the clinical effect of a modified version of constraint-induced movement therapy(MCIMT)on motor recovery in the upper extremities of stroke patients. Methods Twenty-seven stroke patients were randomly divided into a treatment group(n=14)and a routine group(n=13).The Bobath approach and functional arm exercise were administered to the patients in the routine group(2 h/d,5 times a week)for ten weeks.The MCIMT administered to the treatment group involved restriction of the functional arm by a sling during 90%of waking time and during training(by shaping)of the affected extremity(1 hour per day,three times a week)in addition to activities of daily living exercise for at least 2 hours every day.The main outcome measures were the ten items of the simplified test for evaluating hand function(STEF)and Wolf's Motor Function Test(WMFT). Results No significant difference was observed between the 2 groups with regard to WMFT and STEF scores before treatment.After treatment,both groups had improved their WMFT and STEF scores significantly,but there was a significant difference between the treatment and the control groups with regard to WMFT scores and 7 items of the STEF test. Conclusion Modified constraint-induced movement therapy is effective not only for promoting motor recovery of the upper extremity,but also for improving the flexibility and velocity of movement instroke patients.