Objective To investigate the features of HIV-1-specific Cytotoxic T-Lymphocyte (CTL)responses in Chinese.Methods The HIV-1-specific CTL responses were analyzed in an IFN-?ELISPOT assay by using a matrix system containing 820 overlapping peptides spanning the entire HIV-1 Clade B and C consensus sequence.Results The HIV-1-specific CTL response almost clus- tered in Gag and Nef across either HIV-1 Clade B(HIV-1 B)or HIV-1 Clade C(HIV-1 C),while oth- er proteins could also be recognized at different level.In comparison of the response between HIV-1 Clade B and Clade C,the magnitude and frequency was roughly identical with some difference found at single-peptide level.The most frequently recognized peptides of HIV-1 B were located in Nef,GPKEP- FRDYVDRFYKTLR(5/17,29.4%)and Gag,LWVYHTQGYFPDWQNY(5/17,29.4%),while the most frequently recognized peptide of HIV-1 C was located in Gag GPKEPFRDYVDRFFKTLR(6/17, 35.29%).Conclusions HIV-1-specific CTL responses clustered within HIV-1 Gag and Nef in Chinese. However,there was some difference between HIV-1 B and HIV-1 C at single-peptide level.

Objective Preparing platelet rich gel through two-times centrifugal technique and co-culturing chondrocytes with PRG, then observing the proliferation and gene expression of chondrocytes, in order to provide a favorable way to prepare tissue engineering cartilage. Methods Centrifugating venous blood of rabbit through two-times centrifugal technique to obtain platelet rich plasma( PRP) ,then detecting the concentration of various growth factor in PRP. Admixing PRP with chondrocytes of rabbit and activating them with activator. After co-culti-vation,the proliferation of chondrocytes through MTT method and expression of ACAN,CollagenⅡand SOX-9 through realtime-PCR were ob-served,and compared with common cultured chondrocytes. Results The concentrations of PDGF-AB,TGF-β1,IGF-1 and VEGF in PRG were significantly higher than those in blood(P<0. 05). After co-cultivation, the proliferation rate of chondrocytes and the expression of ACAN,Collagen Ⅱ and SOX-9 were significantly higher than that of common cultured chondrocytes(P<0. 05). Conclusion Co-culturing chondrocytes with PRG is able to promote the proliferation and gene expression of chondrocytes. We considered that it is a excellent method to construct tissue engineering cartilage.

Objective To observe the protection effects of sodium ?-aescinate(SA) on the nervous function in the rats with early spinal cord injury(SCI). Methods One hundred and twenty SD rats were randomly divided into four groups(n=30).Rats in the blank control group were performed laminectomy only,while those in the other three groups were injured at the level of Tl1 spinal segment by Allen's weight drop method(10 g ?10 cm) and immediately intraperitoneally given normal saline(5.0 mg/kg)(control group), SA(5.0 mg/kg)(SA group) and methylprednisolone(100 mg/kg)(MP group) once daily,respectively.After 8 h,24 h,96 h,7 d and 14 d,spinal cord function change of posterior limb were determined with Rivlin method.The rats were sacrificed and the injured segments were resected for pathological analysis. Results As time prolonged,the rehabilitation of spinal cord function with various degree could be observed in each group.Function rehabilitation was found among the rats in the control group,SA group and MP group 96 h after injury,and more rehabilitation was gained later in the latter two groups,while that was not the case in the control group.Rats in the SA and MP group gained more significant rehabilitation than those in the control group(P0.05).It was revealed by pathological analysis that no necrotic neurons was found in the blank control group,and the necrotic neurons in the SA group and MP group were significantly less than the control group at the same time points(P

OBJECTIVETo study the inhibitory activity of Aeschynanthus maculatus on alpha-glucosidase.

METHODThe inhibilitory model of in vitro alpha-glucosidase was established. Active extracts of A. maculatus were isolated and identified bymultiple chromatographic methods, and their molecular structures were identifiied by spectral techniques.

RESULTSeven coumpounts were isolated from A. maculatus and isolated as lupeol(1), stigmasterol(2), ursolic acid(3), stigmast-5,22(E)-diene-3beta-ol(4), beta-daucosterol(5), 3-hydroxy-12-taraxasten-28-oic-acid(6) and oleanic acid(7). Compounds 1 (IC50 25.41 mg x L(-1)),3(IC0 4.42 mg L(-1)),4(IC50 11.50 mg x L(-1)),6(IC50 14.17 mg x L(-1)) and 7(IC50 2.88 mg x L(-1)) had higher inhibitory activities than that of acarbose (IC50 1103.01 mg x L(-1)) as the positive control drug.

CONCLUSIONCompound 1-7 were isolated from this plant for the first time. Compound 6 was isolated from Gesneriaceae family for the first time. Compound 7 was isolated from Aeschynanthus genus for the first time.

Limit test of flavones in diterpene ginkgolides meglumine injection materials by UV-Vis and HPLC-DAD method was studied in this essay. The HPLC-DAD method has lower LOD (about 1% of the UV-Vis), that is, the sensitivity is higher than UV-Vis method. Through the analysis of the kinds of flavonoids ingredients in the samples by LC-MS, the three compounds with highest contents are kaempferol, quercetin and isorhamnetin. Kaempferol, quercetin and isorhamnetin were chosen as reference compounds for HPLC analysis, and the HPLC separation analysis was carried on an Agilent Eclipse plus C18 column (4.6 mm x 250 mm, 5 μm) with methanol and water containing 0.4% phosphoric acid (50: 50) as mobile phase, and the flow rate was 1.0 mL x min(-1). The detection wavelength was set at 360 nm. This method has good specificity, precision and reproducibility. The LODs of quercetin, kaempferide and isorhamnetin were 27.6, 22.3, 29.5 μg x L(-1). The average recovery was 87.9% (RSD 3.3%), 91.7% (RSD 3.1%), 88.3 (RSD 1.3%) for quercetin, kaempferide and isorhamnetin, respectively. Based on the 10 batches of sample results and sensitivity of different HPLC, the content of total flavonoids ingredients of diterpene ginkgolides meglumine injection materials was limited no more than 2 x 10(-5). This method is simple, quick and has good maneuverability, and could be used to the limit test of flavonoids in the diterpene ginkgolides meglumine injection materials.
Chromatography, High Pressure Liquid ; methods ; Diterpenes ; analysis ; Drugs, Chinese Herbal ; analysis ; Flavones ; analysis ; Ginkgolides ; analysis ; Limit of Detection ; Mass Spectrometry ; methods

Objective To explore the characteristics of calf muscle movement of the normal volunteer in vivo by velocity encoded phase contrast MRI (VE-PC MRI ).MethodsTwenty four healthy subjects were divided into the young group (30-40 years) and the elderly group (60—78 years).All subjects were positioned on the scanner table and did ankles flexion-extension voluntary movement,the moving images of tibialis anterior muscles (TA),medial gastrocnemius (MG) and soleus muscle (SOL)on the left were scanned with VE-PC MRI.The calf muscles contraction velocity were analyzed by software.The Mann-Whitney U test was used for comparison of calf muscles contraction velocity in vivo between the young group and the elderly group. Results During cyclic ankle flexion-extension,dorsiflexion phase (1—10 phases)in the two groups,TA showed concentric contraction,plantar flexion phase (11—20 phases),MG and SOL did concentric contraction. At 3-6 phases during dorsiflexion,velocity of TA (M value:-37.66,-53.00, -60.66, -56.00 mm/s) in the young group is higher than in the elderly group (M-value:- 30.33, - 42.49, - 53.00, - 48.67 mm/s),at 13—15 phases during plantar flexion,velocity of MG (M-value:- 47.66, - 60.00, - 66.33 mm/s) in the young group showed higher velocity than in the elderly group (M-value:-25.17,-37.99,-50.98 mm/s),at 12—14 phases during plantarflexion,SOL showed higher velocity in the young group (M-value:-27.21, -40.10, -49.13 mm/s) than in the elderly group (M-value:- 20.66,- 29.61,- 37.16 mm/s) ( all P ＜ 0.05).Conclusions During active ankle cyclic flexion-extension movement,velocity of TA,MG and SOL in the elderly group are decreased as compared to the young group. VE-PC MRI can reflect biomechanical characteristics of calf muscle in vivo,and provide a new kind of measurement for lower extremities.

Objectives To evaluate the effects of enteral nutrition(EN)and parenteral nutrition(PN)support in critically ill surgical patients. Methods 80 patients who could not eat were randomized into two groups:one group receivedEN,the other group received PN. The nutrition parameters including nitrogen balance, serum levels of albumin, prealbumin, transferrin and retinal-binding protein,immunity function parameters including immunoglobulin A, immunoglobulin G,immunoglobulin M,and natural kill cell activity, and patients'tolerability parameters including glutamic-oxalacetic transaminase, glutarmic-pyruvic transaminase, blood urea nitrogen and creatinine,and plasma glucose were compared. Results After 10 days'therapy,the two groups all turned from negative nitrogen balance to positive nitrogen balance. The nutrition parameters and immunity function parameters increased significantly in both groups and the latter in group EN increased more. For tolerability parameters, blood sugar in group PN increased obviously and no change in EN group. ConclusionsBoth EN and PN play a rde in nutrition support and elevation of immunity function. Patients tolerate well. EN has better effects on elevation of immunity function and patients tolerate better compared with PN.

ObjectiveTo investigate the relationship between Dicer expression and clinicopathological characteristics and prognosis by detecting the expression of Dicer in hilar cholangiocarcinoma tissues and cells.MethodsThe expression of Dicer in tissues was detected using immunohistochemistry.Western blotting and RT-PCR were used to investigate Dicer expression in QBC939 and HIBEpic cells.The relationship between Dicer expression and clinicopathological characteristics was analyzed.A Kaplan-Maier analysis was performed to analyze the disease-free survival (DFS) and overall survival (OS) after radical surgical resection of hilar cholangiocarcinoma.ResultsWhen compared to control,Dicer was significantly down-regulated in hilar cholangiocarcinoma tissues (P＜0.05) and in QBC939 (P＜0.05).The expression of Dicer was higher in well differentiated adenocarcinoma than poorly and moderately differentiated tumours. Univariate analysis showed low expression of Dicer protein was significantly correlated with short disease-free survival and overall survival of patients with hilar cholangiocarcinoma after radical surgical resection (P＜0.01). Multivariate analysis revealed that the expression of Dicer was the most important factor for predicting prognosis after radical surgical resection of hilar cholangiocarcinoma (P＜0.05).ConclusionsDicer could be used as a prognostic marker for hilar cholangiocarcinoma.

Objective To assess the effects of rehabilitation training on lower extremity muscle function in stroke patients using velocity-encoded phase-contrast MRI ( VE-PC MRI). Methods Twenty-nine stroke patients at the convalescence stage were divided into a treatment group (14 cases) and a control group (13 cases).The subjects performed calf muscles stretches supine in a scanner bed with voluntary ankle flexion-extension movements.Imaging data were collected from the tibialis anterior muscle (TA),the medial head of the gastrocnemius (MG) and the soleus muscle (SOL) of the affected side.The treatment group also underwent rehabilitation including treadmill training,while the control group received only walking training without the treadmill.Barthel index (BI) and Berg balance scale (BBS) scores were measured after rehabilitation. Results Compared with baseline,the contraction velocity of the TA (phases 2 to 7),the MG (phases 12 to 18 ) and the SOL (phases 13 to 16) on the affected side increased significantly in the treatment group.In the control group,the contraction velocity of the calf muscles on the affected side improved,but fewer phases were significant-for the TA phase 4,for the MG phases 13 to 15 and for the SOL phase 15.In both groups,BBS and BI scores improved significantly compared with baseline.The average improvement in the treatment group was significantly greater than that in the control group. Conclusions Rehabilitation including treadmill training can improve calf muscle contraction function on stroke patients' affected side,and also their ADL performance and balance.VE-PC MR1 can provide quantitative in vivo measurements of the contraction function of stroke patients' calf muscles.

AIM: To investigate whether there was nicotinic acetylcholine receptor subunit α 7 (nAChR α 7 ), choline acetyltransferase(ChAT), acetylcholinesterase(AChE) expression and its regulation in mature dendritic cells (DCs). METHODS: Bone marrow(BM) -derived DCs from healthy BALB/c mice were incubated with rmGM -CSF and rmIL-4, and stimulated to mature with LPS. Meanwhile, light microscope and flow cytometry were used to identify DCs, as well as immunocytochemistry, immunofluorescence, flow cytometry and RT - PCR methods were used to dectect expression of nAChR α 7, ChAT and AChE. Flow cytometry was also used to analyze nAChR α 7 expression with mecamylamine (MEC) in 12 h. RESULTS: Both protein and mRNA expression of cholinergic system nAChR α 7, ChAT and AChE were found in mature DCs. Furthermore, nAChR α 7 distributed principally in cell membrane, while ChAT and AChE in cytoplasm. Protein expression of AChE was stronger as compared with ChAT ( P ＜ 0. 05), and there was a trend toward increasing as compared with nAChR α 7. And then, the expression of nAChR α 7 was down regulated by MEC as compared with the group without MEC stimulation(P ＜ 0. 05 ). CONCLUSION: An innate cholinergic system was in mature DCs, which was affected by extrinsic factor ( i. e. , MEC). And it may be involved in anti - inflammation immune adjustion of cholinergic closed - circuit.