Objective To evaluate the effect of intrathecal resveratrol on activation of Ca2+/ calmodulin-dependent protein kinase Ⅱ (CaMK Ⅱ) in spinal dorsal horn neurons of rats with bone cancer pain.Methods Thirty-two adult female Sprague-Dawley rats,weighing 180-220 g,were equally randomized into 4 groups using a random number table:sham operation group (group S),bone cancer pain group (B group),and bone cancer pain + solvent control group (BD group).Walker 256 mammary gland cancer cell suspension (4× 105 cells/ml) 5 μl was injected into the medullary cavity of the right tibia in B,BR and BD groups.Normal saline 5 μl was given in group S.On 12,13 and 14 days after injection of mammary gland cancer cells,resveratrol 200 μg/10 μl was injected intrathecally once a day in group BR,and 1％ dimethyl sulfoxide 10 μl was intrathecally injected once a day in group BD.Before injection of mammary gland cancer cells (T0) and on 3,5,7,10,12 and 14 days after injection of mammary gland cancer cells (T1-6),mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured.The rats were then sacrificed and L4,5 segments of the spinal cord were removed for confirmation of the location of phosphorylated CaMK Ⅱ (p-CaMK Ⅱ) in spinal dorsal horn neurons (by immunofluorescence) and for detection of p-CaMK lⅡ expression (using Western blot).Results Compared with group S,MWT and TWL were significantly decreased at T2-6,and p-CaMK Ⅱ expression was upregulated at T6,and p-CaMK Ⅱ was mainly co-expressed with neurons in B,BR and BD groups.Compared with group B,MWT and TWL were significantly increased at T5,6,and p-CaMK Ⅱ expression was down-regulated at T6 in group BR.There was no significant difference in MWT,TWL,and p-CaMK Ⅱ expression at each time point between group B and group BD.Conclusion Resveratrol can alleviate hyperalgesia in rats with bone cancer pain and inhibited activation of CaMK Ⅱ in spinal dorsal horn neurons may be involved in the mechanism.

AIM:To explore the effect of Pycnogenol on transforming growth factor-β1 ( TGF-β1)-induced he-patic stellate cell activation .METHODS:Cultured LX-2 cells were treated with 5μg/L TGF-β1 and different concentra-tions (0, 10, 25 and 50 mg/L) of Pycnogenol.The viability of the LX-2 cells under the conditions with or without autoph-agy inhibitor 3-MA and ERK inhibitor PD98059 was determined by MTT assay .The protein levels of α-SMA, ColⅠ, TIMP-1, LC3-Ⅱ/Ⅰ, beclin 1, p-ERK1/2 and ERK1/2 were detected by Western blot .RESULTS:Compared with con-trol group, 5μg/L TGF-β1 treatment elevated the cell viability , and increased the protein levels of α-SMA, ColⅠ, TIMP-1, LC3-Ⅱ/Ⅰ, beclin 1, p-ERK1/2, and ERK1/2 in the LX-2 cells (P<0.05).However, these effects were reversed by Pycnogenol pretreatment in a dose-dependent manner and the inhibitory effect of 50 mg/L Pycnogenol was the most sig-nificant in the LX-2 cells (P<0.05).Furthermore, compared with TGF-β1 group, pretreatment with 50 mg/L Pycnog-enol, 5 mmol/L 3-MA or 20 μmol/L PD98059 downregulated TGF-β1-induced cell viability and the protein levels of α-SMA and LC3-Ⅱ/Ⅰ in the LX-2 cells ( P<0.05 ) .CONCLUSION: Pycnogenol suppresses TGF-β1-induced hepatic stellate cell activation via p-ERK and autophagy inhibition .

ObjectiveTo observe the effects of salmeterol fluticasone on the glucose metabolism and bone density of the elderly patients with chronic obstructive pulmonary disease (COPD) combined with type 2 diabetes mellitus (T2DM).MethodsThirty-one patients with COPD combined with T2DM were divided into 2 groups by random digits table,14 cases in control group were given conventional therapy,17 cases in experimental group were given conventional therapy and fluticasone propionate 50/500 μg,twice a day,for 3 months.The fasting blood glucose,postprandial 2 hours glucose,glycosylated hemoglobin,plasma cortisol and bone density respectively before and after treatment were detected.ResultsIn experimental group,the fasting blood glucose,postprandial 2 hours glucose,glycosylated hemoglobin and bone density before treatment was (5.25 ± 0.21 ) mmol/L,(7.14 ± 0.33 ) mmol/L,(5.58 ± 0.26 )％,( 1.96 ± 0.11 ) g/cm2,and after treatment was(5.31 ± 0.27 ) mmol/L,(7.22 ± 0.29 ) mmol/L,(5.67 ± 0.23 )％,(2.03 ± 0.15 ) g/cm2,there was no significant difference (P ＞ 0.05).In control groups,the fasting blood glucose,postprandial 2 hours glucose,glycosylated hemoglobin and bone density before treatment was (5.33 ± 0.35) mmol/L,( 7.26 ± 0.29 ) mmol/L,( 5.62 ± 0.19 )％,( 1.88 ± 0.20 ) g/cm2,and after treatment was ( 5.36 ± 0.31 ) mmol/L,(7.30 ± 0.35 ) mmol/L,( 5.69 ± 0.26 )％,( 1.98 ± 0.17 ) g/cm2,there was no significant difference (P ＞ 0.05 ).There was no significant difference in plasma cortisol before and after treatment in two groups (P ＞0.05).ConclusionInhaling salmeterol fluticasone for elderly patients with COPD combined with T2DM is safe.

Objective To investigate the neuropsychological and functional neuroimaging features in patients with cerebellar infarction (CI).Methods We analyzed 59 CI patients (27 left CI,32 right CI) and 26 healthy control subjects who received standard and experimental cognitive testing and neuroimaging study.We compared the cognitive manifestations between the groups with Student' s t test.Results Patients with CI(left/right) achieved significantly lower scores in auditory verbal learning test (AVLT) of memory test (12.27 ± 1.37 vs 9.33 ± 1.90/10.25 ±2.20,t =6.46,4.26,P ＜0.05),Associative Learning of Clinical Memory Scale (22.77 ± 3.07 vs 18.67 ± 1.98/16.84 ± 3.55,t =5.74,6.69,P ＜ 0.05),symbol digit modalities test (SDMT) of visuospatial test(42.54 ±6.32 vs 20.85 ±9.57/34.84 ± 16.10,t =9.68,2.47,P ＜ 0.05),and errors responses (RE) of Wisconsin card sorting test for executive function (16.77 ± 2.64vs 52.22 ± 16.29/54.47 ± 16.27,t =11.15,12.89,P ＜ 0.05).Patients with left CI had significantly lower scores in correct responses percentage (RCP; 58.71 ± 10.93 vs 78.43 ± 5.26,t =-8.41,P ＜ 0.05)and significantly higher scores in the trials to compete first category (RF; 23.59 ± 9.79 vs 14.12 ± 3.75,t =4.68,P ＜ 0.05).Those finding suggests left CI would cause impairment on abstract conceptualization and concept formation; The patients with right CI had significantly lower scores in total memory quotient (86.69 ± 7.56 vs 112.02 ± 9.70,t =-11.17,P ＜ 0.05),higher scores in perseverative responses (RP ;44.59 ± 17.50 vs 8.23 ± 3.46,t =11.47,P ＜ 0.05) and nonperseverative responses errors percentage (nRPE; 44.00 ±20.67 vs 10.58 ± 2.35,t =9.07,P ＜ 0.05).It means right CI would cause serious deficits on memory,cognitive shift and attention.The fibers between cerebellum and frontal,parietal lobe were reduced in CI patients,compared with healthy control.Conclusions These results suggest that cerebellum participated in the formation of part of cognitive function by connection with cerebrum.After CI,that the fibers contacted with the prefrontal and parietal reduced is the possible mechanisms for cognitive impairment.

Objective: To observe the pathological changes of the lens and anterior lens capsule of the patients with familial congenital aniridia, and discuss the histopathological etiology of the fragility of the anterior capsule and the significance of surgical project. Methods: Anterior lens capsules and lens specimens were obtained from aniridic patients during cataract surgery. The intraoperative behavior of each capsule was noted, after which the specimens were submitted for histopathologic evaluation and electron microscope examination. Results: The anterior lens capsule was extremely fragile and remarkably thin. Degenerative changes(degeneration, necrosis, loss) of the lens epithelium and discontinuity of the lens epithelium were found in some specimens. Proliferation and double layer of the epithelial cells in some area of the specimens can be seen also. Ply structure of the anterior capsule became thin or disappeared. Conclusion: Degenerative or proliferative changes of the lens epithelial cells were associated with the thinness and extreme intraperative fragility of the anterior lens capsules in familial aniridia with cataract. Greater awareness of anterior capsule fragility in some aniridic patients with cataract may reduce the risk of capsule complications and lead to safer surgical outcomes.

BACKGROUND: The transplantation of microencapsulated cell is becoming a hotspot modality in the therapy of Parkinson disease (PD). The application of Alginate-polysysine-alginate (APA) is currently limited due to fragility and pericystic fibrosis although it has been used in clinic. In this study, the native Alginate-chitosan-alginate(ACA)microencapsulated pheochromocytoma cells (PC12 cells) are transplanted into the region of corpus striatum in the injured side of the brain of the PD rat model, the functional recovery of rotational behavior and pathological changes are also observed in the control, sham and treated groups.OBJECTIVE: To observe whether the transplantation of ACA microencapsulated PC12 cells into the brain can improve the rotational behavior in the rat model of PD.DESIGN: Randomized controlled experiment.SETTING: Dalian Research Institute of Physiochemistry, Chinese Academy of Sciences.MATERIALS: Totally 40 adult male Wistar rats with body mass of(220±10) g, ACA microcapsule and PC12 cells were used in this study.METHODS: The experiment was carried out in the animal experimental laboratory of Second Hospital, Jilin University and Dalian Research Institute of Physicochemistry, Chinese Academy of Sciences between May and December 2002. Native ACA were used to microencapsulate the PC12cells. These rats were randomly divided into the following three groups,treated group (10 rats received microencapsulated PC12 cell transplantation), control group (7 rats received unencapsulated PC12 cell transplantation) and sham group (6 rats received empty microencapsule transplantation). The transplantation site was the region of corpus striatum in the injured side of brain. The difference of rotational behavior included by apomorphine was compared before and after the transplantation in these rats,the morphological changes of the transplanted microcapsules and activity of the microencapsulated cells were also detected.MAIN OUTCOME MEASURES: ①Rotational behavior of the rats before and after transplantation. ②Pathological change in the regions of substantia nigra and corpus striatum. ③ The integrality of retrieved microencapsule and the bioactivity of retrieved PC12 cells.RESULTS: ① At the 4th week of transplantation, rotational behavior was significantly decreased in the encapsulated PC12 cells treated group compared with that of the groups received empty microencapsules transplantation [(6.9±2.8),(10.5±1.6) r/min, P ＜ 0.05].Tbis behavioral improvement could last at least three months. Although the unencapsulated PC12 cells also can improve the rotational behavior compared with before transplantation[(5.6±l.1 ), (9.5±1.5) r/min, P ＜ 0.05], which only lasted two months and fetal tumor formed in the skull of some rats. There was no significant difference in rotational behavior of the rats before and after transplantation in the empty microencapsule transplantation group. PC12 cells of retrieved microencapsulate grew well after re-culture, and have bioactivity.CONCLUSION: Transplantation of ACA microencapsulated PC12 cells into the brain can improve can improve the rotational behavior of rat PD model induced by apomorphine. ACA microcapsule can both isolate the host's immune system effectively and prevent the formation of tumor, and have a promising application in clinic.

Sulfonamides (SAs), such as sulfaguanidine (SGD), sulfadiazine (SDZ), sulfathiazole (STZ) and sulfamethazine (SMZ), can drastically inhibit the chemiluminescence (CL) intensities generated in both Ag-Luminol and Ni-Luminol systems.Based on these observations, a novel method of high performance liquid chromatography (HPLC) coupled with CL detection was established.Both the Ag-Luminol and Ni-Luminol CL systems were employed as detectors, and the performances of the two detecting systems were compared.After separated by HPLC, four SAs reacted with Ag-Luminol and Ni-Luminol CL system, respectively.Chromatographic conditions were as follows: reversed-phase C18 column (250 mm × 4.6 mm,5 μm), gradient elution, and 0.1% (V/V) formic acid-methanol as mobile phase with flow rate of 1 mL/min.CL conditions were as follows: [Ag]=1.4×10.-4 mol/L (in 0.12 mol/L NaOH);[Ni]=1.5×10.-5 mol/L (in 0.12 mol/L NaOH);[Luminol]=1.2×10.-7 mol/L;and flow rate=1.0 mL/min.Under the optimal conditions, the detection limits of Ag-Luminol CL system were 0.15, 0.96, 1.10, 1.50 μg/mL for SGD, SDZ, STZ, and SMZ, respectively, and the recovery were 81.0%-101.5%.Comparatively, the detection limits of Ni-Luminol CL system were 1.5, 17.2, 16.8 μg/mL for SGD, SDZ and STZ, and the recoveries was 83.9%-110.8%.The result showed that the Ag-Luminol CL system had a much better performance.The method was applied to the determination of the residues of the above four SAs in milk with satisfactory results.

Aim To prepare a new oral colon-specific delivery formulation and to investigate the release profile in vitro and the colon-specific delivery property in vivo in dogs. Methods Sodium 4-aminosalicylic acid was selected as the model drug. The combination of Eudragit RL30D and RS30D were used as sustained-release film, and Eudragit FS30D used as enteric film, which was expected to release drug depending on pH and time. The release profile of tablets was studied in three phosphate buffers with the pH 6.5, 7.0 or 7.4 for 12 h after a simulated gastric presoak for 2 h in 0.1 mol · L-1 HCl. The tablets were radiolabelled with 99mTc to make their release times and positions in the gastrointestinal tract be followed using a gamma camera. Results For the in vitro study, there was no drug released in 0. 1 mol ·L- 1 HCl for 2 h, and release occurred slowly when pH was above 6.5. Drug was released faster while pH was higher. For the in vivo study, the coated tablets remained intact in the upper gastrointestinal tract, and drug release began after the colonic arrival. The uncoated tablets, however, disintegrated in the stomach of the dogs rapidly. Conclusion The coating could protect the drug until the tablets reached the ascending colon, where drug was released slowly for over 10 h.

A comprehensive analytical method based on ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was developed for the simultaneous determination of active components and anti-diabetic) drugs in propolis health foods. The samples were extracted by ultrasound extraction with methanol). The insoluble residue of extract was removed by freeze-centrifuging. The analysis was performed on an ACQUITY UPLC C_(18)) column(50 mm ×1 mm, i.d., 1.7 μm) utilizing a gradient elution profile and a mobile phase of 0.3% formic acid in water and acetonitrile. The analytes were detected using an electro spray ionization tandem mass spectrometry with multiple reaction monitoring. Qualitative and quantitative analysis was based on the peak area of the parent ion and two fragment ions. The LOD and LOQ level for 14 active components ranged from 0.7 mg/kg to 42.0 mg/kg and 2.2 mg/kg to 140 mg/kg respectively, and recoveries were 77.8%-113.6%, with the intra- and inter-day precision less than 15%. The LOD and LOQ level for 9 anti-diabetics) ranged from 0.1 mg/kg to 0.9 mg/kg and 0.3 mg/kg to 2.5 mg/kg respectively, and recoveries) were 79.3%-108.5%, with the intra- and inter-day precision less than 15%. The method is simple) and sensitive, and can be used for quality control of propolis.