Seven compounds were isolated from the alcohol extract of leaves of Ipomoea batafas Lam,by column chromatography on silica gel. They were identified by physical and chemical methods as quercetin-3-glucoside (Ⅰ), kaempferol-4＇, 7-dimethyl ether(Ⅱ), ombuin (Ⅲ),quercetin (Ⅳ), quercetin-3＇, 4＇, 7-trimethyl ether(Ⅴ), ?-sitosterol (Ⅵ) and sucrose (Ⅶ).All of them were isolated from this plant for the first time.

BACKGROUND:Pressure sensitive instrumented shoes are fast and easily used tools to measure ground reaction forces. Currently researches about the utilities of these systems in assessment of gait in children with neurological diseases have been started to run.OBJECTIVE: To find the gait characteristics of dynamic plantar pressure in children with spastic cerebral palsy. DESIGN: A cross-sectional study.SETTINGS: Department of Rehabilitation, Songgang People's Hospital; Department of Rehabilitation, Shenzhen Children's Hospital; Department of Rehabilitation Medicine, the First Affiliated Hospital of Sun Yat-sen University. PARTICIPANTS: ① Patient group: Twenty children with spastic cerebral palsy were selected from Shenzhen Children's Hospital from May 2004 to April 2005, including 9 boys and 11 girls, aged 26-66 months old, and they all could walk for more than 10 m independently. ② Normal control group: 52 healthy children with normal walking ability were enrolled, including 28 boys.and 24 girls, aged 35-76 months old.METHODS: Ultraflex gait analysis system was used to perform continuous plantar pressure tests of both groups of children. Diagram and data of dynamic plantar pressures in gait were recorded and analyzed with a computer. MAIN OUTCOME MEASURES: ① 10 sequential gait cycles were chosen from the continuous stable steps to calculate the average data with the software; ② Differences of the characteristic parameters of plantar pressure curve between the two groups.RESULTS: ① The curves of plantar pressure to time of testees in the normal control group were of the wave shapes with two peaks and one valley. Half of the CP children could not perform the two-peak shape pressure curve in the gait analysis. One kind of abnormal plantar pressure curve style was a wave shape with only one peak. Another kind of abnormal plantar pressure curve style was a shape showing asymmetry between sides and variation in different steps even in the same side. Two-peak shapes of dynamic plantar pressure were recorded in 10 CP children. ② Characteristic parameters picked up from the curves of plantar pressure had been compared between the two groups. Significant difference have been found in ts of both sides and tz1,tz2,tz3 on the right side between the two groups. It was shown that the single-limb support time of both sides in the CP children was longer than that in healthy children [left: (0.879± 0.658), (0.472±0.051) s; right: (0.818±0.682), (0.478±0.050) s; P ＜ 0.01]. The tz1 and tz3 on the right side in the CP children were longer and while the tz2 was shorter than those in the healthy children. CONCLUSION: ① Using dynamic plantar pressure gait analysis system, plantar pressure curves can be observed directly, and information about kinetic characters can be drawn straightly. ② Part of patients with spastic CP could not perform the two-peak shape pressure curves in gait analysis. In patient can be recorded of two-peak shape dynamic plantar pressure curves, ts of both sides are shorter than those in healthy children, tz1 and tz3 on the right side is longer and tz2 is shorter than those in healthy children.

Aim: To evaluate the safety of glufosfamide in Beagle dogs. Methods: The safety of glufosfamide in Beagle dogs following single iv and multiple iv administration for 3 months was observed. Results: The minimum lethal dose of glufosfamide was 100 mg/kg. The maximum non-lethal dose of glufosfamide was 50 mg/kg. The approximate lethal dose of glufosfamide was 75-100 mg/kg. No serious adverse reactions was observed in Beagle dogs following multiple iv administration once a week for 3 months at the dose of 20 mg/kg, 40 mg/kg, and 60 mg/kg. However, organ injury was observed extensively in the high dose-group and mediumt-dose group. The low-dose group showed a light change in individual organs. Conclusion: Under the experimental conditions, 40 mg/kg and 60 mg/kg iv administration once a week for 3 months can lead to extensive organ injury in Beagle dogs, so the dose shouldn't be higher than 20 mg/kg.

BACKGROUND:To build up an effective method of isolating and culturing granule cels is a pivotal step to enhance fertilization-embryo transfer rate. Current studies mainly focus on the isolation methods of human ovarian granulosa cels rather than cel counting, purity and subsequent growth. OBJECTIVE: To establish the effective methods of isolating, purifying and culturing human ovarian granulosa cels in vitro. METHODS: Folicular fluid was harvested from women undergoing fertilization-embryo transfer procedures. Human ovarian granulosa cels were obtained from the folicular fluid by lysis treatment, precipitation method or density gradient centrifugation. Granulosa cel mucus masses were digested with type I colagen enzyme or hyaluronidase and then cultured in the culture medium with or without autologous folicular fluid. RESULTS AND CONCLUSION: Lysis treatment yielded the largest amount of granulosa cels compared to the precipitation method and density gradient centrifugation (P > 0.05,P < 0.05, respectively). Cels prepared by the three methods showed the same cel viability. After 24 hours of culture, the precipitation method obtained the largest amount of adherent granulosa cels (P < 0.05); and the density gradient centrifugation obtained the least amount of cels (P < 0.05). Compared with type I colagen enzyme, hyaluronidase took less time to digest the cels thoroughly. Autologous folicular fluid could promote the growth and survival of granulosa cels. These findings indicate that the precipitation method, though time-consuming, can obtain the highest cel viability and harvested the largest amount of granulosa cels after culture; hyaluronidase is more suitable for digesting granulosa cel mucus mass than type I colagen enzyme; autologous folicular fluid added into the culture medium is more conducive to granulosa cel growth.

Objective To examine the reliability and similarity of amplitude of low-frequency fluctuation (ALFF)for different scan duration.Methods 10 min 10 s resting state fMRI data acquired from 13 healthy volunteers separately were divided into 10 pieces of the same length.The reliability and similarity of ALFF were assessed for each scan length (1-10 min).Results Spatial maps of ALFF steadily appeared in each time period (1-10 min);but during the time evolution,both spatial distribution and extent of ALFF increased gradually.At only one minute,there were detectable statistical differences within the group.When the time ran-ging from 5 to 10 min,relatively stable ALFF results in the group achieved.Conclusion ALFF is a relatively stable index,it can be easily affected by scan duration.In the case of 1-5 min,it should be carefully analyzed and interpretated particularly.

AIM:To investigate whether KCNE 2 participates in the development of pathological hypertrophy .METHODS:Bidirectional ma-nipulations of KCNE2 expression were performed by adenoviral overexpression of KCNE 2 or knockdown of KCNE2 with RNA interfer-ence in PE-induced neonatal rat ventricular myocytes .Then overexpression of KCNE 2 in mouse model of left ventricular hypertrophy in-duced by transverse aortic constriction (TAC) by ultrasound microbubble-mediated gene transfer were used to detect the therapeutic function of KCNE2 in the development of hypertrophy .RESULTS:KCNE2 expression was significantly decreased in PE-induced hy-pertrophic cardiomyocytes and in hypertrophic hearts produced by TAC .Knockdown of KCNE2 in cardiomyocytes reproduced hypertro-phy, whereas overexpression of KCNE2 attenuated PE-induced cardiomyocyte hypertrophy .Knockdown of KCNE2 increased calcineurin activity and nuclear NFAT protein level , and pretreatment with nifedipine or FK 506 attenuated decreased KCNE 2-induced cardiomyo-cyte hypertrophy .Overexpression of KCNE 2 in heart by ultrasound microbubble-mediated gene transfer suppressed the development of hypertrophy and activation of calcineurin-NFAT and MAPK pathways in TAC mice .CONCLUSION:These findings demonstrate that cardiac KCNE2 expression is decreased and contributes to the development of hypertrophy via activation of calcineurin -NFAT and
MAPK pathways .

Objective To study whether serum matrix metalloproteinase 9, serum transforming growth factor ? 1, serum type C IV collagen concentrations are alternatoin in various stages of patients with T2DM. Methods 64 cases T2DM patients were divided into three groups according to urine albumin excretion rate (UAER):normoalbuminuria (NA)group and microalbuminuria (MA) group and overt diabetic nephropathy (ODN)group. Serum MMP 9,TGF ? 1 and C IV were detected in all cases. Results Serum MMP 9 in MA and ODN groups were higher than NA and normal control groups ( P

Objective To study neuromechanism for uremic restless syndrome (URLS),functional MRI (fMRI) analysis and were used to explor main activity.Methods Resting state functional magnetic resonance imaging (MRI) was performed on 29 patients with uremic restless legs syndrome and 25 healthy controls.The values of the regional homogeneity (ReHo) of the two groups were calculated and analyzed statistically.Result As compared with the control group,patients with uremic restless legs syndrome showed emotional and mental abnormality.Increased ReHo values in bilateral supplementary motor area,bilateral thalamus,left insular lobe,left hippocampus (P＜0.05),and decreased ReHo values in anterior cingulate gyrus,bilateral posterior central gyrus,right amygdala were found in patients with uremic restless legs syndrome (P＜0.05).The severity score of RLS was respectively positively correlated with dialysis duration (r =0.57,P =0.002),PTH level (r =0.419,P =0.033)BAI (r =0.528,P=0.006),and BDI (r =0.567,P =0.003).Conclusion Neuronal activity in related brain area in patients with uremic restless legs syndrome were found.This abnormality provides an objective diagnostic basis for the explanation of restless legs syndrome in maintenance hemodialysis patients.

Objective To assess the value of 14 common food allergens specific IgG in children with chronic digestive diseases.Methods The specific allergen food IgG was measured by utilizing enzyme-linked immunosorbent assay(ELISA) in 69 children with chronic digestive diseases,who had chronic abdominal pain and recurrent vomiting above three months with no organic disorders,incurable chronic diarrhea and anorexia with failing to thrive.Results The food allergen specific IgG increased was detected in 68 cases.The positive cases were 98.55%.There was one to nigh kind of food allergen specific IgG increased in 68 patients.Positive above two kind of specific IgG was 86.96%.The most specific IgG was to egg 92.8% and milk 73.9%;the other rates were: wheat 40.6%,soybean 27.5%.Chicken and pork were negative.All the patients changed their daily diet according to the assay results.More than 58.46% cases improved with their symptoms significantly in four weeks,about 26.15% cases improved to some extent.15.38% cases did not improved.After followed four months,there were 70.70% cases improved with their symptoms significantly,18.46% cases improved to some extent,only 10.76% cases no improved.Conclusion The detection of 14 common food allergens specific IgG might be significant valuable in assessment and treatment children with chronic digestive diseases.

ObjectiveTo reproduce a clinically relevant two-hit model of sepsis complicated by pneumonia and to explore the correlation between two-hit and immune state.Methods Eighty-one male Sprague-Dawley ( SD ) rats were divided into groups according to the random number table. Forty-five male rats were assigned respectively to sepsis-alone group, pneumonia 4 days and 7 days after sepsis groups, respectively. Survival rate of each group was observed. Another group of 36 male rats were divided into normal control group, sepsis-alone for 1, 4 and 7 days groups, and sepsis complicated by pneumonia for 4 days and 7 days after sepsis groups, each group consisted of 6 rats. Cecal ligation and puncture (CLP) was done in rats, andStreptococcus pneumoniae suspension (bacteria count 1×1010 cfu/mL) was injected via the nose on the 4th day or 7th day after CLP. Rats were sacrificed at corresponding time points, and 1 day after challenge ofStreptococcus pneumoniae on the 4 days or 7 days post CLP for the collection of blood and tissue samples to make bacterial count of the blood, splenocyte count, biochemical indices, cytokines concentration, pathological changes in spleen and apoptotic cells.Results① Compared with the rats of sepsis-alone group, the rats in pneumonia 4 days after CLP group had poor survival rate (4 vs. 11,χ2 = 6.533,P = 0.011), while no difference was found between pneumonia 7 days after CLP group and sepsis-alone group (9 vs. 11,χ2 = 0.600,P = 0.439).② The blood bacterial count and all the biochemical indexes were sharply increased on 1 day post-CLP in the rats of sepsis-alone group, and then they gradually lowered. Compared with the rats of 1 day post-CLP, the proportion of splenocytes were decreased on the 4th day post-CLP [dendritic cells (DC): (0.69±0.09)% vs. (0.87±0.31)%, CD4+T cells: (21.05±2.89)% vs. (24.84±4.59)%, CD8+ T cells: (10.62±1.79)% vs. (13.40±1.31)%, allP< 0.05], but T-regulatory cell (Treg) count was higher on the 4th day after CLP compared with sepsis-alone rats [(3.14±0.74 )% vs. (2.87±1.08)%,P< 0.05]. The biochemical indices, including alanine transaminase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and serum creatinine (SCr) were obviously lowered on 7 days post-CLP compared with 1 day after CLP [ALT (U/L): 35.33±11.52 vs. 81.00±38.40, AST (U/L): 70.33±42.16 vs. 156.00±28.11, BUN (mmol/L): 5.30±2.27 vs. 9.13±4.04, SCr (μmol/L): 55.33±10.67 vs. 96.67±45.79, allP< 0.05]. The serum levels of tumor necrosis factor-α (TNF-α) and interleukins (IL-6, IL-1β) peaked on the 1st day after CLP [TNF-α:(18.03±2.88) ng/L, IL-6: (10.37±4.20) ng/L, IL-1β: (102.44±51.46) ng/L], and high mobility group box-1 (HMGB1) peaked on the 4th day after CLP [(1.76±0.71)μg/L]. The levels of anti-inflammatory cytokines transforming growth factor-β1 (TGF-β1 ) and soluble tumor necrosis factor receptor-Ⅰ (sTNFR-Ⅰ) maintained at high levels [7 days post-CLP: TGF-β1 was (0.90±0.56) ng/L, sTNFR-Ⅰ was (1.56±0.39) ng/L]. The spleen pathology became more marked with the time in the group of sepsis-alone, meanwhile the number of apoptotic spleencytes increased 4 days post-CLP as compared with that of the 1st day post-CLP (cells/HP: 52.99±20.79 vs. 16.05±3.28,P< 0.05).③ Compared with the same period of sepsis-alone group, the rats with pneumonia 4 days post-CLP group showed a higher blood bacterial count (log cfu/mL: 1.78±0.54 vs. 0.25±0.18,P< 0.05), while no difference was found between 7-day of post-CLP pneumonia group and sepsis-alone group (log cfu/mL: 0.57±0.46 vs. 0.13±0.12,P> 0.05). The same trend of changes, with slight reduction in splenocytes and biochemical indices were found between the groups of sepsis followed by pneumonia and sepsis-alone, but no significant difference was found. The level of HMGB1 in the 4-day group of sepsis with complication of pneumonia was further decreased compared with sepsis-alone group (μg/L:1.17±0.74 vs. 1.76±0.71,P< 0.05), and IL-1β in the 7-day group of sepsis complicated pneumonia was further higher than those of sepsis-alone group in the same period (ng/L: 105.73±25.06 vs. 61.04±31.29,P< 0.05), while there were no differences in levels of other cytokines between two-hit group and sepsis-alone group. Apoptosis of spleencytes in the 4-day group of sepsis complicated pneumonia was more marked than that of sepsis-alone group at the same period (cells/HP: 74.48±22.47 vs. 52.99±20.79,P< 0.05), while no difference was found between the 7-day groups of sepsis complicated pneumonia and the sepsis-alone group (cells/HP: 28.70±4.13 vs. 30.43±14.55, P> 0.05).Conclusions The mortality of this two-hit model with complication of pneumonia 4 days after CLP was significantly higher than that of single sepsis model. The ability of bacteria clearance was decreased, and immunocyte apoptosis was exacerbated. These findings may be with the result of the occurrence of immunoparalysis in the mid stage of sepsis. The two-hit model reproduced on 7 days after CLP might suggest reconstruction of host immune function, and maybe associated with the recovery of immune response.