Animals ; Arginine ; pharmacology ; therapeutic use ; Female ; Male ; Myocardial Ischemia ; drug therapy ; pathology ; physiopathology ; Myocardial Reperfusion Injury ; drug therapy ; pathology ; physiopathology ; Rabbits

·AIM: To investigate whether Erigeron Breviscapus (vant) Hand-Mazz (EBHM) EBHM has neuroprotective effect against N-methyl-D-aspartate (NMDA)-induced neuron death in retinal ganglion cell layer (RGCL).· METHODS: 60 healthy SD rats were randomly divided into four groups. 6 animals were normal control group (group A). The others were divided as group B (EBHM group), group C (normal saline+NMDA group) and group D (EBHM + NMDA group). Each group had 18 rats.10nmol NMDA was intravitreally injected to induce partial damage of the neurons in RGCL in the right eyes of Groups C and D. Same volume PBS was intravitreally injected into the left eyes as self-control. Groups B and D were pre-treated intraperitoneally with 6g/L EBHM solution at a dose of 150mg/kg body weight/day seven days before and after NMDA treatment. Group C were administrated intraperitoneally with 9g/L normal saline at the same time of EBHM injection. Rats were sacrificed at 4,7,14d after NMDA treatment. Flat whole retinas were stained with 5g/L cresyl violet and neuron counting in RGCL from both eyes were observed. Each subgroup had 6 rats.· RESULTS: There was no significant difference of neuron counting in RGCL between the right eye and the left eye in group A (P=0.200). There was no significant difference between normal control group and EBHM group either in the right eyes or in the left eyes at 4, 7 and 14 d respectively after intravitreal injection of 10nmol NMDA in group C and group D. (P=0.636, P=0.193). Neuron counting of RGCL in group C and D was significantly decreased in the NMDA-treated eyes at 4, 7 and 14d after intravitreal injection (P＜0.001). There was no significant difference between self-control eyes group and normai control group(P＞0.05). However, neuron counting was significantly higher in the EBHM+NMDA group than normal saline +NMDA group at 14days after intravitreal injection (P=0.044), but was lowered than normal control group (P＜0.05).· CONCLUSION: EBHM has no effect on neuron counting of RGCL when administered alone in normal rats.The results indicates that EBHM plays a partial protective role in NMDA-induced neuron loss in RGCL in the rats.

BACKGROUND: It has been proved that platelet activation is involved in the development of diabetic angiopathy. Glycoprotein (GP) Ib/IX/V complex is one of the main platelet membrane glycoproteins, and the receptor of both von Willebrand Factor and thrombin. It plays a key role in the process of platelet activation.OBJECTIVE: To observe the expression changes of GP Ib/IX/V complex and its component GP Ibα in patients with type 2 diabetes mellitus. DESIGN: Case-control study. SETTING: Department of Integrated Chinese and Western Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.PARTICIPANTS: A total of 51 type 2 diabetic outpatients who visited Union Hospital were enrolled from December 2005 to January 2007. The diagnosis was based on the independent criteria from WHO in 1999. Of all the 51 patients, there were 23 females and 28 males, with a peripheral platelet count of over 50×109 L-1. All the subjects had no history of administrating drugs two weeks before the examination, which would potentially influence platelet count. According to disease controlling condition, the patients were assigned to well controlled patient (WCP) group (n = 25) and poorly controlled patient (PCP) group (n =26); and according to whether angiopathy was accompanied, diabetic patients were divided into vascular disease (VD) group (n =27) and non-vascular disease (NVD) group (n =24). Meanwhile 23 healthy subjects were enrolled as normal control group. Informed consents were obtained from subjects and their relatives. The experiments were approved by the ethical committee of Union Hospital.METHODS: Fasting venous blood was harvested from all the subjects' elbow on the early morning of visit day.①Biochemical analysis: Fasting plasma glucose (FPG), glycosylated hemoglobin (HbA1c) and fasting plasma insulin (FINS) was measured by the Clinical Chemistry Department in Union Hospital.② Measurement of platelet membrane GP Ib/IX/V complex and its component GP Ibα: First, 3 mL cubital fasting blood was drawn from each subject and was anti-coagulated with 38 g/L natrium citricum. After that, all samples were fixed with 10 g/L paraform for 45 minutes. Then 50 μL well-fixed blood was added into the polystyrene tube, meanwhile 20 μL monoclonal antibody, such as CD42a-b-c-d and PE-labeled CD42b, was respectively mixed gently with the blood sample and incubated at room temperature in dark for 30 minutes. Next, 20 μL FITC-labeled rat IgG was mixed with the sample containing CD42a-b-c-d and incubated equally. In the end all blood samples were analyzed by FACS420 flow cytometry and the results were expressed as mean fluorescence intensity (MFI). ③ Platelet maximun aggregation rate (MAR) was detected according to reference.MAIN OUTCOME MEASURES: ①Biochemical indicators;②The expressions of GP Ib/IX/V complex and GP Ibα;③Platelet MAR.RESULTS: Fifty-one patients with type 2 diabetes mellitus and twenty-three healthy subjects were all involved in the result analysis.①There were significant differences in FINS in WCP group and PCP group compared with normal controls (P ＜ 0.01). FPG and HbA1c were significantly higher in PCP group compared with normal control group and WCP group (P ＜ 0.01).②Expressions of GP Ib/IX/V complex and GP Ibα were significantly lower in WCP group and PCP group compared with normal control group (P ＜ 0.01), significantly lower in PCP group than in WCP group (P ＜ 0.05), and also significantly lower in VD group and NVD group compared with normal control group (P ＜ 0.01). Moreover the expression of GP Ibα in VD group and NVD group was significantly lower than that of normal control group (P ＜ 0.05), and it also significantly decreased in VD group compared with NVD group (P ＜ 0.05). MFI of GP Ib/IX/V complex had an obvious negative correlation with FBG, HbA1c and FINS (r =-0.634, -0.573, -0.649, P ＜ 0.05), and GP Ibα MFI was obviously negatively correlated with FBG and HbA1c (r =-0.602, -0.543, P ＜ 0.05).③Platelet MAR of diabetic patients were remarkably higher than in healthy subjects (t =-3.852, P ＜ 0.01). Platelet MAR in PCP and VD groups were respectively higher than those in WCP and NVD groups (P ＜ 0.05). CONCLUSION: Platelet activation exists in the early stage of type 2 diabetes mellitus without diabetic angiopathy, and is more obvious after diabetic angiopathy. There is a positive correlation between platelet activation and blood glucose. As a receptor of thrombin and von Willebrand Factor, GP Ib/IX/V complex may be involved in the development of diabetic angiopathy.

OBJECTIVETo elucidate the long-term effect of repeated febrile convulsions (FC) on seizure susceptibility in immature rats.

METHODSWarm-water-immersion rat FC model was developed with SD rats 22 days of age, 15 attacks of seizures were induced in the rats every other day, and some of the rats were left un-stimulated for 3 months, then were re-stimulated. Seizure phenomenon was observed, including the latency and the duration of seizures and the temperature of rats. Hippocampal neuron loss and mossy fiber sprouting were detected by thionine staining and Timm staining.

RESULTSAfter the 15th bath, the latency, the duration of seizures, and the temperature of rats were respectively (4.3 +/- 0.8) min, (5.5 +/- 2.9) min, (42.2 +/- 0.7) degrees C. Three months later, on re-stimulation, in 14 of 19 rats with previous FC experience seizures occurred while in only one of 13 non-FC rats seizure occurred and lasted for 8.5 min. Three months later, the latency, the duration of seizures, and the temperature of rats were respectively (5.4 +/- 0.6) min, (19.3 +/- 5.1) min, and (42.4 +/- 0.4) degrees C (4 rats with status epileptics were not included). The incidence of seizures on re-stimulation in rats of FC group (74%) was significantly higher than that in non-FC group (8%) (chi(2) = 13.50, P < 0.01), and the duration of seizures [(19.3 +/- 5.1) min] was significantly longer than those induced in early life [(5.5 +/- 2.9) min] (t = 10.49, P < 0.01). After the 15th bath, no significant change was demonstrated in rats of different groups. While 3 months later, prominent neuron loss was observed in hippocampal CA(3) region in rats with previous FC experience (P < 0.01). Significant mossy fiber sprouting phenomenon was detected after the 15th bath and 3 months later (P < 0.05).

CONCLUSIONRepeated FC in early life enhances long term susceptibility of rats to seizure.

Animals ; Disease Models, Animal ; Disease Susceptibility ; Hippocampus ; pathology ; Rats ; Rats, Sprague-Dawley ; Seizures ; pathology ; Seizures, Febrile ; pathology ; Time Factors

OBJECTIVETo observe and analyze the pathohistological characteristics of capsules which formed around the mammary prosthesis with different contents. And to provide the selective basis for ideal and safe prosthesis in clinical practice.

METHODS20 specimen of the capsules were taken from 20 cases who receive the operation of prothesis removal for different reasons. HE, Masson and Mallory staining were used to analyse the tissue structure and characteristics under the light microscope.

RESULTSThe common structure including the collagen fibers accumulation, inflammatory cells infiltration and the capillary hyperplasia were found in all specimen. A layer of squamous epithelium-like cell was detected in some specimen. The specific characteristics were also found in different capsules formed around different prosthesis. In the capsules around vegetable oil prosthesis, there was excessive collagen fiber accumulation, and the capsules were much thicker. In the PVP (polyvinylpyrolidone) prosthesis capsules, there was severe inflammatory cell infiltration, and the number of eosinophilic granulocyte increased obviously. In the silicone gel and saline prosthesis capsule, the collagen fibers were well-arranged and the inflammatory cells were much less. Synovial metaplasia was detected in two cases.

CONCLUSION1. The capsules form around the prosthesis in all cases after mammary augmentation. 2. There will be synovial metaplasis in some cases, for vegetable oil prosthesis, the collagen over-accumulated which lead the capsules become thicker and harder. So it is not a kind of ideal mammary prothesis. 4. The severe infiltration of the inflammatory cells especially the large quantity of eosinophilic granulocyte indicate the possibility of the delayed hypersensitive reaction mediated by eosinophilic granulocyte. Cautious attitude should be taken during application.

Adult ; Breast ; pathology ; Breast Implants ; Female ; Humans ; Middle Aged

OBJECTIVETo investigate the polymorphism of DYS287 among 28 ethnic populations in 9 provinces of China.

METHODYAP element was detected by Touchdown PCR amplification and 2% agarose gel electrophoresis.

RESULTSYAP+ frequencies in these ethnic populations were as follows: Zang 36.7%, Tu 23.8%, Yi 18.4%, Pumi 11.3%, Tajik 7.4%, Bai 6.7%, Jino 5.1%, Shandong Han 4%, Mulao 2.7%, and Maonan 1.3%. The rest ethnic populations in our study, including Gansu Han, Yunnan Han, Zhuangzu, Daizu, Lizu, Nuzu, Lisu, Naxi, Lahu, Dulong, Hani, Shezu, Weiwuer, Sala, Kerkizi, Dongxiang, Vazu, and Korea didn't carry YAP + element.

CONCLUSIONSZangzu, Tuzu, Yizu, Pumi, Jino, and Baizu, which belong to Sino-Tibetan language family, carry a high YAP + frequency. Sala, Tuzu, and Tajik, regarded as Central Asia by origin in history and linguistics, also have a high YAP + frequency. Mulao and Maonan, which origin from "Baiyue" ancient ethnic groups, also have a considerable YAP + frequency.

Alu Elements ; genetics ; Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Chromosomes, Human, Y ; genetics ; Electrophoresis, Agar Gel ; Gene Frequency ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic

OBJECTIVETo investigate the frequencies of HLA-Alu repeat polymorphisms (AluMICB, AluTF, AluHJ, AluHG and AluHF) in Chinese Lisu and Nu ethnic populations.

METHODSThe frequencies of HLA-Alu repeat polymorphisms in above populations were determined with polymerase chain reaction (PCR). The associations between HLA-Alu repeat polymorphisms and HLA-A, HLA-B and HLA-C alleles were also analyzed. Phylogenetic trees were constructed with genetic distance calculated from the frequencies of HLA-Alu repeat polymorphisms.

RESULTSFrequencies of AluTF*2 and AluHF*2 were different between the two populations (P< 0.05), while those of other three insertions were similar. The strength of association between HLA-Alus and HLA alleles were different (P< 0.05) in the two populations. Although AluMICB*2 were associated with HLA-B*56:01 in both populations, the association was stronger in Lisu population (74.0%) but moderate in Nu population (30.7%). HLA-Alus were associated with particular HLA subtypes, e.g., AluHG*2 with certain HLA-A*02 subtypes. By phylogenetic analysis, Lisu and Nu were clustered together with southern Chinese and Thai populations.

CONCLUSIONThe distribution of HLA-Alus and the strength of associations between HLA-Alus and HLA class I alleles have varied between the two populations. Study of this association may facilitate identification of origins, evolution, progenitor haplotypes and recombination within the HLA class I region.

Adolescent ; Adult ; Aged ; Alleles ; Alu Elements ; Asian Continental Ancestry Group ; genetics ; Child ; Female ; Genes, MHC Class I ; Humans ; Male ; Middle Aged ; Phylogeny ; Polymorphism, Genetic ; Young Adult

OBJECTIVETo evaluate the impact of specific siRNA on survivin gene in transfected leukemia cells.

METHODThe small interfering RNA (siRNA) targeted survivin mRNA was synthesized in vitro and was transfected into K562 cell by Hiperfect into human leukemia cell line K562, which has high survivin expression level. The level of survivin mRNA expression was determined by quantitative reverse transcription polymerase chain reaction (RT-PCR) with SYBR GREEN I. The apoptosis index of cytotrophoblasts were determined and analyzed by FCM (Annexin V-FITC/PI staining methods). The cell proliferation was examined by MTT at 48 h and 72 h after transfection.

RESULTThe level of mRNA expression was significantly inhibited by the siRNA 48 h and 72 h after transfection, the suppression rate of survivin mRNA separately reached 85.21%, 94.35% mensurated by quantitative RT-PCR with SYBR GREEN I, cell proliferation was inhibited significantly by 45.02% and 50.88%, respectively, the apoptotic rate detected by Annexin V-FITC assay reached 12.28%and 21.55%, respectively.

CONCLUSIONThe chemosynthesized siRNA targeting survivin could significantly down-regulate survivin mRNA. Survivin siRNA was able to inhibit the proliferation of leukemia cell line K562. Survivin may become a new target for leukemia gene therapy.

Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Gene Silencing ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; K562 Cells ; RNA, Small Interfering ; pharmacology ; Transfection

This study was purposed to explore the mechanisms of drug resistance of HL-60/ADR cells and to compare the reversal drug-resistance effects of antisense oligonucleotides (AS ODN) of XIAP (X-linked inhibitor of apoptosis protein) and AS ODNs of MRP (multidrug resistance-associated protein) by use alone or in combination. Reverse transcription-PCR and Western blot were applied to detect the expression of XIAP, BCL-2, MRP and MDR1 in mRNA and protein levels of HL-60 cells and HL-60/ADR cells, respectively. Fully phosphorothioated AS ODN of XIAP and MRP was delivered into HL-60/ADR cells with Lipofectamine 2000 in the form of liposome-ODN complexes alone or in combination. CCK-8 cell viability assay was used to determine the effect of AS ODN of XIAP and MRP used alone or in combination on the chemotherapy sensitivity of HL-60/ADR cells to daunorubicin (DNR). Reverse transcription-PCR and Western blot were applied to examine the changes of XIAP, MRP in mRNA and protein levels respectively. The results showed that MRP and XIAP were both significantly higher in HL-60/ADR cells than those in HL-60 cells. AS ODN of XIAP and MRP down-regulated the expression of XIAP and MRP in HL-60/ADR cells and increased the sensitivity of HL-60/ADR cells to DNR, respectively. AS ODN of XIAP + MRP did not enhance the inhibition expression of XIAP in HL-60/ADR cells but increased the sensitivity of HL-60/ADR cells to DNR significantly as compared with AS ODN of XIAP (P < 0.05). AS ODN of XIAP + MRP did not increase the concentration of DNR nor enhanced the inhibition expression of MRP in HL-60/ADR cells but increased the sensitivity of HL-60/ADR cells to DNR significantly (P < 0.05), as compared with AS ODN of MRP. It is concluded that both XIAP and MRP may be involved in the drug resistance mechanisms of HL-60/ADR cells. Drug-resistance of HL-60/ADR cells can be reversed significantly when antisense oligonucleotides of XIAP and MRP were used in combination.
Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; HL-60 Cells ; Humans ; Multidrug Resistance-Associated Proteins ; biosynthesis ; genetics ; Oligonucleotides, Antisense ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; X-Linked Inhibitor of Apoptosis Protein ; biosynthesis ; genetics

OBJECTIVETo investigate the inhibitory effect of RNA interference on chronic myeloid leukemia (CML) bcr/abl oncogene expression.

METHODSThe small interference RNAs (siRNAs) were synthesized in vitro. K562 cells stably expressing bcr/abl gene were transfected with the siRNA by electroporation, both the non-transfected cells and non-specific siRNAs transfected cells were taken as controls. The enhanced green fluorescent protein (EGFP) plasmid was used as positive control and the transfection efficiency was detected by flow cytometry. Inhibitory effect of siRNAs was demonstrated by real-time quantitative RT-PCR and Western blots. Cell proliferation was measured by MTT assay and apoptosis by Annexin V-FITC assay.

RESULTSThe transfection efficiency was about 70%. The synthesized siRNAs inhibited CML bcr/abl oncogene expression at both mRNA and protein levels. siRNAs could inhibit K562 cell proliferation to 47% and 56% at 24 h and 48 h after transfection, respectively, and induce cell apoptosis from 1.00% in control group to 15.05% and 19.4% at 24 h and 48 h respectively.

CONCLUSIONAt the cell level, inhibition of CML bcr/abl oncogene expression by chemically synthesized siRNAs provides the new method for anti-leukemia study.

Apoptosis ; genetics ; Cell Proliferation ; Fusion Proteins, bcr-abl ; genetics ; Humans ; K562 Cells ; RNA, Small Interfering ; Transfection