A rat monoclonal antibody of the IgG1 class, McAb B5A8, specific for the cAMP-dependent protein kinase(PKA) was produced. Western blot analysis revealed specific binding of the antibody to protein of 52-56 kd.The affinity-purified McAb BSA8 was labeled with FITC. Immunofluorescent localization of PKA was examined in human fibroblasts, gastric cancer cell line (MGc 80-3)and EAC cells. The distribution of PKA on the cytoplasmic microtubule network, Golgi region and nucleoli were observed. PKA was localized in the nuclear region in G2 phase of synchronized MGc 80-3 cells, and it was only found around the nuclei of MGc 80-3 cells which were incubated with DBcAMP. The changes in the distribution of PKA in cells are discussed.

Synchronous cells in different phase (G_1, S, G_2, M) of cell cycle were obtained from human gastric low-differentiated mucous adenocarcinoma cells (MGC 80-3), which werec ultured with nitrous oxide under high pressure and blocked with overdosage of TdR. Dynamic changes of the mitochondria stained with Rhodamine-123 and succinate dehydrogenase demonstrated by cytochemical method were observed in various phase of the cells at 0, 24, 36, 60 hours after treatment with HPD plus red light. The results showed that mitochonodria of all four phases are of impairment immediately after photoradiation, SDH reactivity is decreased slightly at 24 hours, the activities of SDH is the weakest. As time goes on, we observed that mitochondria gradually recovered to its original structure and then SDH returned to its normal level. The recovery rate of mitochondria and SDH was in the following order, i. e. S, G_1, G_2, andM. The relationship between these changes and cell killing is briefly discussed.

Objective To evaluate whether susceptibility weighted imaging (SWI) can be used in definition of penumbra during acute stage of cerebral infarction,compared with perfusion weighted imaging (PWI).Methods Ischemic stroke patients within 3 days after onset were included.They adopted multimodal magnetic resonance imaging examination,including regular magnetic resonance imaging sequence (T1 WI,T2 WI and T2-weight fast fluid-attenuated inversion-recovery),diffusion weighted imaging (DWI),PWI and SWI.Alberta Stroke Programme Early CT Score was done on DWI,SWI and PWI.The mismatch of SWI-DWI (minimal indensity projection (mIP)-DWI) was compared with that of PWI-DWI (mean transit time (MTT)-DWI) and analyzed statistically.The application of prominent vein (PV) on SWI as a sort of alternation of cerebral blood volume (CBV) and direct observation of thrombosis in arteries on SWI were done.Results The SWI-DWI (2.39 ± 1.42) and the MTT-DWI (2.72 ± 1.49) mismatch showed no statistically significant difference (r =0.726,P ＞ 0.05).The grade of PV was positively related with the CBV of the ipsilateral brain tissue on admission (r =0.564,P ＜ 0.05).SWI showed the similar ability with magnetic resonance angiography to judge responsible blood vessels with susceptibility vessel sign.Conclusion SWI-DWI can evaluate the ischemic penumbra.PV may reflect the increased blood volume of the lesion side of the brain tissue.SWI can reveal the thrombosis of the responsible vessels.

Objective To investigate the clinical effect and feasibility of interventional treatment of cervical artery stenosis in the elderly patients aged 75 years and over. Methods The data of 60cases aged 75 years or over who underwent interventional treatment of the cervical arteriostenosis involving carotid artery (CA), vertebral artery (VA) or proximal segment of the subclavian artery (SCA) were analyzed retrospectively. The clinical manifestations, imaging characteristics,interventional managements and follow-up results were recorded. Results In this cohort, the mean age was (78. 9±3.7) years (range from 75 to 89). The 50 patients (93.3%) complained of cerebral ischemic symptoms, and all the patients had concurrent diseases or risk factors, including hypertension, diabetes mellitus, coronary heart disease (CHD), stroke history, and so on. Digital subtraction angiography (DSA) data showed 55 cases (91.7%) had 2 or more cerebral arteries with a stenosis exceeding 30%. Among all cases, 84 lesions were treated with 84 stents, with a technical success rate of 98.8%. After stenting, the percent diameter stenosis of lesions decreased from a mean of (80.8 ± 12.9) % to (7.1 ± 9.5 ) %. The periprocedural and 30-day postoperative neurological complication rate was 8. 3 %, resulting in a permanent complication rate of 5 %. Clinical improvement rate was 87.5%. During a follow-up period of (36.7±26. 3) months (range from 5 to 99), there were 4 deaths: 2 died from myocardial infarction, 1 died from brain metastases of lung cancer and 1 died from cerebral hemorrhage. Cerebral infarction recurred in 3 cases. Imaging follow-up in 78. 3% of patients for 66 stents, including ultrasound, CTA, MRA or DSA, showed that the general in-stent restenosis rate was 9. 1%, and the restenosis rate of VA, CA and SCA was 21.7% (5/23), 2.6%and 0, respectively. Conclusions The results of this series suggest that interventional treatment ofcervical artery stenosis in the elderly patients aged 75 years and over is effective and feasible. In our experience, clinical comprehensive management and skillful technique of the operator are equally important for the elderly patients aged 75 years and over with high incidence of concurrent diseases or risk factors.

OBJECTIVETo investigate antitumor constituents from n-butyl alcohol extract of Alternanthera philoxeroides.

METHODThe constituents were isolated with silica gel, gel permeation chromatography, and purified by HPLC. Their structures were elucidated by spectroscopy and acid hydrolysis. The antitumor effects of extracts and isolated compounds were tested by MTH method in vitro.

RESULTSeven compounds were isolated and elucidated as followings: oleanolic acid 3-O-beta-D-glucuronopyranoside (1), oleanolic acid 28-O-beta-D-glucopyranoside (2), oleanolic acid 3-O-beta-D-glucuronopyranoside-6'-O-methyl ester (3), chikusetsusaponin IV a methyl ester (4), hederagenin 3-O-beta-D-glucuronopyranoside-6'-O-methyl ester (5), 4,5-dihydroblumenol (6), 6S,7E,9R-6,9-di-hydroxymegastigma-4,7-dien-3-one-9-O-beta-D-glucopyranoside (7). Compound 1 showed significant inhibitory effect against Hela and L929 with inhibitive ratios 91.3% and 92.9% at 30 mg x L(-1), respectively.

CONCLUSIONCompounds 4, 5 and 7 were isolated from this genus for the first time. Compound 1 showed significant inhibitory effect against Hela and L929 at 30 mg x L(-1).

Amaranthaceae ; chemistry ; Antineoplastic Agents ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Humans ; Hydrolysis

In order to improve quality of molecular pharmacognosy teaching, the open experiment is applied. Under the guidance of tutors, students conduct the whole experiment independently. Students' abilities of independent thinking and comprehensive-experimental conduction were enhanced in the open experiment. Meanwhile, the authors discuss the problems of open experiment and propose some reflection and suggestions.
Education, Medical, Undergraduate ; economics ; manpower ; methods ; standards ; Humans ; Pharmacognosy ; education