BACKGROUND: Astragalus root can inhibit apoptosis through reducing the release and interstitial accumulation of excitatory amino acids, alleviating calcium overloading and antioxidative effect.OBJECTIVE: Astragalus root was used to treat anoxic-ischemic brain injury in immature brain. We evaluated the effect of astragalus root on caspase-3 mRNA expression, and meanwhile, labyrinth test was employed to investigate the intervention of astragalus root on learning and memory function of mature rats after anoxic-ischemic brain injury.DESIGN: Randomized and controlled study.SETTING: Pediatric Department, Zhongda Hospital Affiliated to the Medical College of Southeast University; Pathological Department, the Basic Medical Sciences Institute of Southeast University.MATERIALS: From October 2002 to June 2003, this study was conducted at the Experiment Center of the Medical College, Southeast University.A batch of 114 seven-day-old SD rats were selected from the same brood and divided into 3 groups, namely, sham-operation group (n=18), model group (n=48) and astragalus root group (n=48). Astragalus injection was produced by Chengdu DIAO Pharmaceutical Factory, with 10 mL astragalus injection corresponding to 20 g raw material.METHODS: Animal model of anoxic-ischemic brain injury was established in model group and astragalus root group, but was not established in sham-operation group. In astragalus root group, immediately after establishing anoxic-ischemic model and at the same time point each day, 0.08 mL astragalus injection was administered intraperitoneally until the 7th postoperative day. In model group, 0.08 mL normal saline was administered at the same time points. In sham-operation group, no treatment was given. In astragalus root group and model group, animals were decollatedat 24 hours and 5 days postoperatively to take out the brains. In sham-operation group,animals were decollated and their brains were taken out at 24 hours postoperatively. In all the groups, hippocampal brain injury was detected using histopathological method combined with semi-quantified RT-PCR methods for detecting caspase-3 mRNA. Adult rats aged 90 days were used in modified y maze to examine their learning and memory functions. All these three experiments were independent.MAIN OUTCOME MEASURES:① Hippocampal brain injury in each group was evaluated using pathological method.② Caspase-3 mRNA in the ligated side of hippocampus was detected.③ Results of modified Y maze test were analyzed.RESULTS:All of the 114 rats entered the statistical analysis.① Assessment ofhippocampal brain injury in each group with pathological method:In sham-operation group, the bilateral hippocampus showed no swelling or necrosis, and neural cells in this area had normal morphological features with a density of (87.7±0.6) × 103 per high amplification field. In model group, the ligated side of hippocampus was swollen with a widened spatium and the cell density decreased to (68.8±3.0) × 103 per high amplification field, which significantly differed from that in sham-operation group (P ＜ 0.01). At the fifth day, the volume of ligated side of hippocampus reduced with pyramid layer disorganized and neural cells sparse at a density of (48.7±2.2) × 103 per high amplification field. These changes were significantly different from those of sham-operation group and the same side at 24 hours (P ＜ 0.01). At 24 hours the ligated side of hippocampus was less swollen in astragalus root group than in model group.At day 5, the whole hippocampus was observed. At these two time points,cell death rate in astragalus root group was significant lower than that in model group(P ＜ 0.01).②Caspase-3 mRNA in the ligated side of hippocampus in all the groups: In sham-operation group, the expression of caspase-3 was low, with an absorbency value of 0.220±0.009. In model group, after ischemia and anoxia its expression increased. At 6 hours, it was 11% higher than that in sham-operation group. In astragalus root group, mRNA level reached its peak, which was 260% higher than that in sham-operation group (P ＜ 0.01). The peak of mRNA continued, decreased after 48 hours and returned to baseline at 5 days and 7 days. The fluctuation of mRNA was similar between astragalus root group and model group,but the peak value at 24 hours and 48 hours in astragalus root group was 44％-46％ lower than that in model group (P ＜ 0.01). ③ Results of modified Y maze test: As compared to model group, in astragalus root group, the number of training times for meeting the standard made by the Association was significantly smaller [(45.7±2.7), (16.1±2.5) times, P ＜ 0.01] and at 24 hours after anoxia and ischemia, memory retention was significantly higher [(48.3±11.7), (80.0±9.0)%, P ＜ 0.01].CONCLUSION: Astragalus root can effectively inhibit the apoptosis of neural cells in hippocampus in immature brain after anoxia and ischemia and enhance the survival rate of them. This protective effect may be related to its inhibitory effect on the expression of caspase-3. Meanwhile, astragalus root can dramatically improve learning and memory function of the immature brain after anoxia and ischemia.

The dialysis method was employed to prepare blank and doxorubicin (DOX) loaded micelles formed by temperature- and pH- sensitive polyhistidine-co-polyDL-lactide-co-glycolide-co-polyethyleneglycol-co-polyDL-lactide-co-glycolide-co-polyhistidine (PHis-b-PLGA-b-PEG-b-PLGA-b-PHis). The critical micelle concentrations (CMC) of the copolymers were measured with Pyrene Fluorescent Probe Technique. The temperature- and pH- sensitive properties of the blank micelles solution were investigated by optical transmittance measurement. The morphology and diameter of DOX micelles were characterized by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The entrapment rate and drug-loading rate were determined with dialysis method. The in vitro release study was further performed to examine the temperature- and pH-responsive drug release behavior from DOX-loaded micelles. The results indicated that the CMC, entrapment efficiency and drug-loaded amount of the micelles were 7.5 x 10(-3) g x L(-1), 85.2 +/- 3.1% and 10.4 +/- 4.5%, respectively. The DOX micelle was globular-shaped with a mean diameter of 91.1 +/- 15.8 nm. The transmittance of micelle solution consistently increased with the increasing temperature or decreasing pH. In comparison to the drug release profile at physiological conditions (37 degrees C, pH 7.4), the DOX-loaded micelles showed faster drug release rate at higher temperature (41 degrees C), lower pH (pH 7.0, pH 6.5, pH 5.0) or higher temperature and lower pH (41 degrees C, pH 5.0). This indicated that the micelles showed a temperature and pH-triggered drug release pattern. Base on the above results, it can be concluded that PHis-b-PLGA-b-PEG-b-PLGA-b-PHis block copolymer micelles which respond to temperature and pH stimuli are promising smart carriers for anti-tumor drugs with the advantages of temperature- and pH- triggered drug release.

Objective To observe the effects of different quantities of cake-separated moxibustion on blood cells and immunoglobulin in immunosuppressive rabbits; To study the possible mechanism of quantity-effect difference. Methods Immunosuppressive rabbit model was established by intraperitoneal injection of cyclophosphamide. Rabbits were randomly divided into blank group, model group, cake-separated moxibustion 3 Zhuang group, 5 Zhuang group, 7 Zhuang group.Liuwei Dihuang Decoction was used to make herbal-cake, and was put on Shenque (RN8), Guanyuan (RN4), and other acupoints, once for every other day, 10 times in total. Venous blood was taken on the next day after finishing treatment, and the contents of blood cells series and IgG, IgM, complement C3 and complement C4 were detected.Results Compared with blank group, the levels of WBC, NEU, RBC, HGB, HCT, PLT, IgG, IgM, and complement C3 of model group significantly decreased, while the levels of LYM and MONO significantly increased (P<0.01). Compared with model group, the increase of HCT and IgM in the cake-separated moxibustion 3 Zhuang group was not significant, the levels of WBC, NEU, RBC, HGB, HCT, PLT, IgG, IgM, and complement C3 in all treatment groups significantly increased, and LYM and MONO significantly decreased (P<0.01). Compared with cake-separated moxibustion 3 Zhuang group, the levels of WBC, IgG, and complement C3 of cake-separated moxibustion 5 Zhuang group significantly increased, and the levels of WBC, HGB and complement C3 of cake-separated moxibustion 7 Zhuang group significantly increased (P<0.01).Conclusion Cake-separated moxibustion can improve the immune function under immunosuppressive state induced by cyclophosphamide, and different quantities have differences in efficacy.

Phosphoethanolamine N-methyltransferase (PEAMT) is a key enzyme that catalyzes the synthesis of phosphocholine, which is an important precursor of phosphatidylcholine and glycine betaine. A 1249bp 5'-flanking region of phosphoethanolamine N-methyltransferase gene was isolated by anchored PCR, based on the cDNA sequence of PEAMT from halophyte Salicornia europaea. The transcription start site was identified as A and localized at 301bp upstream of the ATG according to the results of RLM-RACE. In SePEAMT promoter region, many potential cis-acting elements were predicted by PlantCARE and PLACE programs. Aside from the basal transcriptional elements TATA-box and CAAT-box, some stress-responsive motifs such as ABRE, HSE and LTR were found. In addition, some pollen-specific activation-related elements were also present in this region. Binary expression vector was constructed by fusing SePEAMT promoter with GUS gene and designated as pPro. The pPro was transferred into tobacco by Agrobacterium-medicated transformation and transient GUS expression analysis indicated that SePEAMT promoter could drive strong GUS expression.

AIM: To explore the expression of caspase -3 (cysteinyl aspartate-specific proteinase) in the neonatal rat cerebral cortex a nd hippocampal after hypoxia-ischemia. METHODS: Sham and hypoxia-ischemia (HI) groups were set up. The neonatal HI procedure was performed in 7-day-old rat pups. The double-lateral co rtex and hippocampal was subjected to pathological assessment, immunohistochemic al staining with caspase-3 antibody and half-quantitative reverse transcription and polymerization chain reaction (RT-PCR) to measure the change in caspase-3 pr otein and mRNA expression. RESULTS: Caspase-3 mRNA in ipsilateral cerebral cortex and hippo campal increased immediately after HI followed by a partial recovery. Thereafter caspase-3 mRNA and protein simultaneously increased with a maximum reached at 2 4-48 h after HI. CONCLUSION: Caspase-3 may play a key role in the development of apoptotic hypoxic-ischemic brain damage (HIBD) in immature rats. Neuroprotective medicine should be used before 24-48 h after HI.

Objective:To improve the quality standard of Tongsai Yinao oral liquids for the quality control. Methods: TLC was used for the qualitative identification of Rhizoma corydails, Rhizoma chuanxiong and Rhizoma acori tatarinowii. HPLC was used to sim-ultaneously determine the content of total ferulic acid in the oral liquids. The determination was performed on a Lichrospher-C18 (250 mm×4.6 mm,5 μm)column with the mobile phase of methanol-0.1% H3PO4(30∶70). The flow rate was 1.0 ml·min-1,the col-umn temperature was at 30℃, the detection wavelength was 321nm and the injection volume was 20μl. Results:The spots on the TLC plates were clear without the interference of negative control. The linear range of ferulic acid was obtained between 2. 24 and 35. 84 μg ·ml-1(r=0. 999 9), and the average recovery was 102. 54%(RSD=0. 85%, n=6). Conclusion:The improved method is accu-rate and feasible. It can be used very conveniently in the quality control.

OBJECTIVETo study the mechanism of injury of cortical nerve cell in the newborn with hypoxia/ischemia brain damage (HIBD), and the neuroprotective effect of Radix Astragali (RA).

METHODSNeonatal HIBD model rats were established and divided into the sham group, the model group and the RA group. Brain of rats obtained at different time points after HIBD to conduct histopathological examination, neuron death rate count, as well as determination of caspase-3 (cysteinyl aspartate-specific proteinase-3) protein mRNA expression in cerebral cortex by immunohistochemistry, semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) respectively.

RESULTSIn the model group, caspase-3 mRNA and protein showed an increase at 6 hrs, reached the peak at 24 hrs, and decreased at 48 hrs after HIBD, on the 5th and 7th day restored to baseline level. After being treated by RA, the neuron death rate of ligated side was obviously reduced, caspase-3 mRNA and protein expression peak value decreased by 45% (mRNA) and 40% - 43% (protein).

CONCLUSIONRA shows markedly neuron protection in immature brain cortex after HIBD, which is related with the inhibition on caspase-3 expression.

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Astragalus membranaceus ; Caspase 3 ; Caspases ; biosynthesis ; genetics ; Cell Survival ; Cerebral Cortex ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Female ; Hypoxia-Ischemia, Brain ; metabolism ; pathology ; Male ; Neurons ; pathology ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley

Objective To compare the gene expression difference between 0.1 and 5 Gy X-ray irradiated cells,and to explore its possible mechanism.Methods A cDNA microarray corresponding to 45033 human genes was used to analyze the transcriptional profiles of normal human lymphoblastoid AHH-1 cells at 4 h after 0.1 or 5 Gy irradiation.The genes with a fold change ≥ 2.0 were identified as the differentially expressed genes.real-lime PCR and Western blot were used to confirm the expression of PERP.Results The microarray assay showed that there were 760 up-regulated genes and 1222 down-regulated genes in the cells at 0.1 Gy,while there were 744 genes down-regulated and 457 genes up-regulated in the cells at 5 Gy.In addition,55 genes were commonly up-regulated and 339 genes commonly down-regulated at 0.1 and 5 Gy.The predominant biological processes of the differential genes responding to low-dose radiation include cell-cell signaling transduction and DNA damage response,and the altered genes after 5 Gy irradiation were related to cell proliferation,differentiation,and apoptosis.Moreover,the expression of PERP gene was down regulated,which was consistent with the data of microarrey assay.Conclusions The quantitative and qualitative differences in the gene expressions may contribute to the diversed biological effects induced by low or high doses of ionizina radiation.

50 adult male SD rats were divided randomly into 5 groups:A,B,C1 ,C2,C3(n =1 0).The rats in roup A was used as a blank controls.The rats in group B and C were with LPS induced periodontitis,those in group C1 ,C2 and C3 received 0.2 ml of baicalin daily injection(0.01 ,0.1 and 1 .0 μg/ml respectively)into the gingival sulcus of the teeth with periodontitis for 3 days.The rats were sacri-ficed 7 days after treatment and periodontal tissues of the related teeth were observed by histology.In group C1 ,C2 and C3 the periodontal inflammation was significantly slighter than that in group B,the osteoclasts count was as following:B >C1 >C2 >C3 >A(P <0.05).The study suggests that baicalin can inhibit destructive effect of LPS to periodontal tissue of rats.

Cutaneous Fistula ; Digestive System Abnormalities ; therapy ; Endoscopy, Gastrointestinal ; methods ; Female ; Humans ; Infant ; Treatment Outcome