Objective To investigate the changes in endothelial progenitor cells (EPCs) number in peripheral blood of patients with acute cerebral infarction and after Naoxintong treatment.Methods Sixty patients were selected as subject and randomly divided into aspirin group and aspirin (30 patients) + Naoxintong group (30 patients).Meanwhile 30 patients without cerebral infarction were served as control group The number of peripheral blood EPCs were detected by flow cytometry at different time point.NIH-NINDS stroke score was used to elevate the neurological function.Results Compared with the control group,number of peripheral blood EPCs significantly decreased in the early stage of acute cerebral infarction (P ＜ 0.05),and then gradually increased until 7th day,which was back to the normal level.There was a positive correlation between improvement of NIHSS and number of peripheral blood EPCs in acute cerebral infarction.Compared with aspirin group,the number of peripheral blood EPCs in Naoxintong group increased sigrificanfly [41.40 ±0.18/million cells vs 41.40 ±0.18/million cells] at 14th day in patients treated with aspirin and Naoxintong.Conclusion The number of peripheral blood EPCs showed a U shape dynamic change in acute cerebral infarctior.Increase the number of peripheral blood EPCs might improve prognosis in patients with acute cerebral infarction.On the basis of routine treatment of aspirin,Naoxintong plus aspirin treatment might improve the number of peripheral blood EPCs in acute cerebral infarction.

Objective To investigate the effects of levodopa/benserazide-loaded poly-lactide-coglycolide (PLGA) microspheres on motor deficits and levodopa-induced dyskinesia in a rat model of Parkinson' s disease (PD) and to explore the mechanisms underlying this effects.Methods The content of levodopa/benserazide released from the microspheres was determined by high-performance liquid chromatography.The rat model of PD was induced by 6-OHDA injections.Then the valid PD rats were treated with levodopa ( 12 mg/kg,s.c.)/benserazide ( 15 mg/kg,s.c.) or microspheres.Forepaw adjusting steps were measured on 1,4,7,10 and 14 days after treatment.After 2 weeks of treatment,the abnormal involuntary movements (AIM) were measured.Phosphorylated dopamine,cAMP-regulated phosphoprotein of 32 kDa (DARPP-32) at threonine 34 levels were determined by immunohistochemistry and Western blot respectively.In addition,the levels of △FosB were measured by Western blot.Results In vivo release test showed that 76.2％ of levodopa and 83.6％ benserazide were released from the microspheres on day 7.Forepaw adjusting steps showed that the scores of forepaw adjusting steps in microspheres-treated PD rats were ( 5.8 ± 1.6 ) and ( 5.2 ± 1.5 ) respectively on 10 and 14 days of treatment,which were increased compared to levodopa-treaded PD rats (2.4 ± 1.1 and 1.2 ± 0.5 ; t =4.12,5.43,all P ＜0.01 ).The AIM scores of microspheres-treated rats ( 16.0 ±2.1 ) were decreased significantly compared to levodopa-treated rats ( 26.0 ± 3.2 ) on day 14 ( t =6.59,P ＜ 0.01 ).Immunohistochemistry indicated that the phosphorylated levels of DARPP-32 in microspheres-treated rats ( (3.7 ± 1.3 ) × 104 )were decreased significantly compared to levodopa-treated rats ( (7.9 ± 2.2) × 104 ; t =2.95,P ＜ 0.05 ).In addition,Western blot showed that the levels of phosphorylated DARPP-32 and △FosB were 119.4％ ± 11.3％ and 149.3％ ± 12.3％ respectively,which were decreased significantly compared to levodopatreated rats ( 184.8％ ± 13.7％ and 300.4％ ± 14.2％ ; t =4.12,2.91,all P ＜ 0.05 ).Conclusions Microspheres can be used to improve the motor deficits and reduce the expression of dyskinesia in PD rats.This may be due to the continuous release of levodopa/beserazide from the microspheres,which leads to continuous stimulation of PD rats and reduces the levels of phosphorylated DARPP-32 and △FosB in the striata of these rats.

OBJECTIVE: To investigate the curative effect of combined transplantation of bone marrow and umbilical cord blood of same sibling in children with β-thalassemia major.METHODS: Eight thalassemia major patients undergoing combined transplantation of bone marrow and umbilical cord blood of same sibling aged from 4.0 to 7.5 years, 5 boys and 3 girls, were recruited at the Department of Pediatrics, Nanfang Hospital,Southem Medical University from January 2005 to March 2009. The patients were classified into three classes according to Pesarothalassamia classification, class Ⅰ to class Ⅱ 7 cases and class Ⅲ 1 case. Donors ranged 1-4 years received 10 μg/kg per day of subcutaneous granulocyte colony-stimulating factor (G-CSF) for 5 consecutive days. Bone marrow was harvested on the fifth day. Bone marrow and umbilical cord blood of the same sibling then were transfused into the patient.RESULTS: Recovery of hematopoiesis was gained in all patients 4 weeks following transplantation. Seven patients suffered from infection of different degree. Four patients developed mild venous occlusive disease. Two patients developed grade Ⅰ acute graft-versus-host disease (GVHD), and one developed grade Ⅰ chronic GVHD. Seven patients were alive and one died of pulmonary infection and heart failure 32 days following transplantation.CONCLUSION: Combined transplantation of granulocyte colony-stimulating factor primed bone marrow and umbilical cord blood of same sibling in children with β-thalassemia major is safe and effective with promising results. However, complications should be paid high attention following transplantation.

This study was aimed to isolate and characterize of chemical constituents in a traditional Mongolian medicine HERBA CLEMATIS. Normal and reverse phase coloumn chromatography, gel filtration chromatography sephadex LH-20, and preparative HPLC were used for isolation and purification compounds from the water extraction of the arial part ofC. aethusaefoliaTurcz. The planar structures and spatial configurations of isolated compounds were identified by high resolution MS, 1D and 2D NMR, and other spectrographic methods. The chemical research on the Mongolian medicine results 6 compounds, dihydrodehydrodiconiferyl alcohol (1), syringaresinol (2), pinoresinol (3), epi-pinoresinol (4), lirioresinol B dimethyl ether (5) and loliolide (6). All the compounds were isolated from this plant for the first time.

Mesenchymal stem cells (MSC) are the non-hematopoietic stem cells with a multi-differentiation potentials, which has a low immunogenicity and immune regulation ability. MSC immune regulation ability is particularly important, such as MSC can inhibit the activation and proliferation of T, B lymphocytes, NK cells and dendritic cells (DC). Meanwhile, MSC is able to reconstruct the human hematopoietic microenvironment, improving the successful rate of hematopoietic stem cell transplantation. Graft versus host disease (GVHD) is the main factor causing hematopoietic stem cell transplantation-related mortality. Based on the above mentioned properties, MSCs are used to treat autoimmune diseases and GVHD, recently. Therefore, deep exploration of the cellular immune mechanisms of MSC to treat GVHD is particularly important. This review focuses on progress of research related to treatment of GVHD by MSC immune mechanisms and briefly summarizes the status of the clinical studies.
Graft vs Host Disease ; immunology ; therapy ; Humans ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; immunology

OBJECTIVETo explore the role of estrogen in the regulation of the expression of claudin-6 and biological behavior in MCF-7 cells.

METHODSRT-PCR and immunocytochemistry were conducted to analyze the expression and localization of claudin-6 in MCF-7 cells treated with 17β-estradiol. CCK-8 kit assay and Scratch Test were conducted to analyze the capability of proliferation and migration of 17β-estradiol treated MCF-7 cells.

RESULTSRT-PCR analysis and immunocytochemistry showed that 17β-estradiol induced a concentration-and time-dependent effect on claudin-6. At 5 nmol/L and at 24 h, 17β-estradiol treatment led to an increased level of claudin-6, which was located in the membrane of MCF-7 cells. CCK-8 analysis showed a significant decrease in the capability of proliferation of MCF-7 cells compared with the control group (P < 0.05). Cells Scratch Test showed decreased migration capability of MCF-7 cells compared with the control group (P > 0.05).

CONCLUSIONS17β-E2 might regulate the expression of claudin-6 and inhibit the proliferation and migration of MCF-7 cells. The inhibitory effects of 17β-E2 on growth and migration of MCF-7 cells may be mediated by claudin-6 expression regulation.

Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Claudins ; Dose-Response Relationship, Drug ; Estradiol ; administration & dosage ; pharmacology ; Female ; Humans ; Membrane Proteins ; metabolism

The effect of glycogen synthase kinase 3beta (GSK3beta) has been repeatedly implicated in cell proliferation, but studies on the effect of GSK3beta in different cell lines with different stimuli have drawn different conclusions. To investigate the direct effect of GSK3beta on cell growth in human lung adenocarcinoma cell line A549, we changed its activity by transient transfection with two kinds of GSK3beta mutant plasmids, constitutively active form S9A-GSK3beta and dominant negative form KM-GSK3beta. Twenty-four hours later, cell counting, flow cytometry and Western blot detection were made respectively. The results showed that enhancing GSK3beta activity caused a decrease in cell number, as well as a higher percentage of cells at G(1) phase. Further, the expression of cyclin D1 was down-regulated by GSK3beta. Taken together, our observations suggest that GSK3beta may induce G(1) cell cycle arrest in a cyclin D1-dependent fashion and therefore possibly plays a growth-inhibitory role in A549 cells.
Adenocarcinoma ; pathology ; Cell Cycle Checkpoints ; Cell Line, Tumor ; Cell Proliferation ; Cyclin D1 ; metabolism ; Down-Regulation ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Humans ; Lung Neoplasms ; pathology ; Transfection

To investigate the roles of glycogen synthase kinase 3beta (GSK3beta) and adenomatous polyposis coli (APC) protein in wound repair of airway epithelial cells (AECs), we established a wound model of airway epithelium in vitro. Then the following tests were undertaken: (1) Western blot was used to detect the levels of total GSK3beta and phosphorylated GSK3beta in human bronchial epithelial (16HBE) cells; (2) The localizations of APC protein was observed by using immunofluorescence technique; (3) Immunoprecipitation was used to investigate the relationship between APC protein and GSK3beta during the repair of 16HBE cells. The results were as follows: (1) The level of phosphorylated GSK3beta increased 0.5 h after scratching (P<0.05), reached a maximum at 6 h (P<0.05), and maintained until 12 h, while the total level of GSK3beta remained constant; (2) Results of immunofluorescence study showed that APC protein clustered with tubulin in the region of the migrating leading cells 6 h after scratching, which was dissimilar with that in the cells 0 h after scratching; (3) GSK3beta and APC protein were immunoprecipitated and analysed on SDS-PAGE. We found that GSK3beta and APC protein were precipitated, indicating that the two proteins existed in a complex. After scratching, dissociation of the two proteins occurred. Taken together, we conclude that scratching caused a decrease in phosphorylation of GSK3beta, and that reduced phosphorylation of GSK3beta promoted APC protein to bind to the plus ends of microtubules and stabilize the growing ends. These observations suggest that APC protein and GSK3beta may synergistically play an important role in the repair of airway epithelium.
Adenomatous Polyposis Coli Protein ; physiology ; Bronchi ; cytology ; injuries ; Cell Line ; Epithelial Cells ; metabolism ; pathology ; Glycogen Synthase Kinase 3 ; physiology ; Glycogen Synthase Kinase 3 beta ; Humans ; Phosphorylation ; Wound Healing ; physiology

To investigate the roles of adenomatous polyposis coli (APC) protein and glycogen synthase kinase 3beta (GSK3beta) of smoking murine model in the repair of the injured airway epithelial cells (AECs) in different stages, 30 male Kun-Ming mice were randomly divided into two groups, the control group and the smoking group. There were 24 mice in smoking group, and 6 animals were separately killed at the end of the 1st, 4th, 8th and 12th week after smoking. Then the following tests were undertaken: (1) HE staining of lung section to observe the morphological changes of the bronchi in the smoking mice. (2) Immunohistochemical staining of APC protein and GSK3beta in the AECs. (3) Western blot was used to detect the levels of APC protein, GSK3beta and phosphorated GSK3beta (p-GSK3beta) in pulmonary tissue. (4) Observing the localizations of APC protein and GSK3beta in the AECs by immunofluorescence technique. The results showed: (1) AECs showed changes of predominant injury (1-, 4-week), repair (8-week) and reinjury (12-week) along with smoking time prolonged. The experimental results indicated that the model of smoking mice was duplicated successfully. (2) Immunohistochemical results showed that the expression of APC protein in the AECs increased after 1-week smoking (0.458 +/- 0.062 vs 0.399 +/- 0.060, P< 0.05 vs control), but was significantly decreased at the end of the 4th week (0.339+/- 0.056, P<0.01 vs control) and increased at the end of the 8th and 12th week (0.387 +/- 0.041, 0.378 +/- 0.037, P<0.05 vs 4-week). The expression of GSK3beta in the AECs of smoking mice obviously decreased (P<0.01 or P<0.05 vs control). (3) Western blot showed that the expressions of APC protein and GSK3beta in lung tissue were consistent with the results of immunohistochemistry; and the levels of p-GSK3beta in all smoking models were higher than that in control. (4) The results of immunofluorescence showed that APC protein was localized mainly near the regions of epithelial cell membrane at the end of the 1st and 8th week after smoking, which were dissimilar with the localization in control, and this change was not seen in the location of GSK3beta. Taken together, these results demonstrate that the expressions and localizations of APC protein, GSK3beta and the activity of GSK3beta are dynamically changed in the AECs with experimental smoking injury at different phases, suggesting that APC protein and GSK3beta may be involved in the regulation of migration and proliferation of AECs, and play an important role in the process of repair of airway epithelium injury.
Adenomatous Polyposis Coli Protein ; metabolism ; Animals ; Bronchi ; pathology ; physiology ; Female ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Lung ; pathology ; physiology ; Male ; Mice ; Regeneration ; Smoke ; adverse effects ; Tobacco ; adverse effects

BACKGROUNDTo find out the timing of serologic responses after illness onset and distribution of IgG antibody to SARS-CoV in SARS cases of transmission chain or non-transmission chain.

METHODSThe IgG and IgM antibodies to SARS-CoV were tested by indirect ELISA in serum samples from 301 clinically diagnosed SARS cases.

RESULTSTotally 158 SARS cases were involved in 15 chains of transmission. The positive rates of SARS-CoV IgG in those chains were 85.70%-100.00% and the overall rate was 94.30% (149/158). The chain of transmission could spread to four generations, but the SARS cases were reduced with increase of generations. There was no significant difference among positive rates of SARS-CoV IgG for generations, Chi square=5.11, P greater than 0.05. The positive rate of SARS-CoV IgG in cases who were not in chain of transmission was 12.59%(18/143) which was statistically significantly different from that of cases in chain of transmission, Chi square=199.64, P less than 0.001. During days 0-7,8-14,15-21,22-30 after onset, the cumulated positive rate of SARS-CoV IgG was 16.67%, 40.00%, 70.00% and 93.10%, respectively, then was kept at the level above 90% and lasted for 217 days. The cumulated positive rate of SARS-CoV IgM during days 0-7 after onset was the same to that of IgG. During days 8-14, 55.17% of cases had seroconversion for IgM which reached a peak (86.96%) during days 21-30. Then the rate rapidly declined.

CONCLUSIONMore than 94% of cases with SARS could produce IgG antibody when they were infected by SARS-CoV. Detecting SARS-CoV IgG could provide a diagnostic evidence for case confirmation. SARS-CoV IgG appeared as early as 7 days after onset and reached the peak at about weeks 4. Then the high rate of antibody was maintained for more than 6 months.

Antibodies, Viral ; blood ; Disease Transmission, Infectious ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; SARS Virus ; immunology ; Severe Acute Respiratory Syndrome ; immunology ; transmission