Post-stroke fatigue (PoSF) is an independent pathologic symptom after stroke which negatively influenced activities of daily living (ADL) and quality of life (QOL) of the stroke patients. PoSF is irrelevant to depression, age, state of the illness, etc. PoSF should be assessed appropriately and alleviated by medicine, diet, sleep, exercise, and health education, etc.

Objective To explore the relationship between reduction of regional cerebral blood flow in the normal appearing white matter (NAWM) and the extent of age-related white matter lesions (WML).Methods We used Fazekas scale to divide all participants into four groups (normal,mild,moderate,severe) according to the extent of the lesions showed on MRI.Regional cerebral blood flow (rCBF) in the area of WML and NAWM was measured by xenon contrast CT examination.Results A total of 56 cases were selected.The average rCBF (ml · 100 g-1 · min-1) in the lesions (WML) around ventricle,in right centrum ovale and in left centrum ovale respectively was 20.33 ± 2.52,21.27 ± 1.02,21.03 ± 1.83 for mild; 16.33 ±2.03,15.55 ±1.71,15.91 ±0.98 for moderate; 14.05 ±2.63,14.46 ±2.17,14.23 ± 1.95 for severe.The average rCBF (ml · 100 g-1 · min-1) in the NAWM around ventricle,in right centrum ovale and in left centrum ovale respectively was 20.79 ± 2.78,22.26 ± 1.9,22.15 ± 2.4 for normal; 21.12 ± 2.95,22.17 ± 1.50,22.25 ± 2.13 for mild,18.02 ± 2.41,19.45 ± 1.94,19.62 ± 1.54for moderate; 16.38 ± 3.22,18.18 ± 2.84,16.74 ± 2.97 for severe.The decrease of rCBF in the severe and moderate lesion areas was more serious than that in the mild lesion areas and reached statistic significance (P ＜ 0.05) ; The decrease of rCBF in the severe and moderate lesion areas was more serious than that in the area of NAWM in the same grade and reached statistic significance (P ＜ 0.05).The decrease of rCBF in the area of NAWM around severe and moderate lesion areas was more serious compared with that around mild lesion areas or normal areas (P ＜ 0.05) ; But the difference of rCBF in the area of NAWM around mild lesion areas and normal areas did not show any statistic significance.Conclusions Chronic ischemia was found to be existed not only in the lesions (WML) but also in the area of NAWM around the lesions,and was related to the extent of the lesions.Chronic ischemia may play a key role in the mechanism of aged-related WML.

In order to reveal the cause of disease occurred in the process of Coptis chinensis growth, this paper studied the bacterial species diversity index of different aged rhizospheric and non-rhizospheric soil planting normal or sick C. chinensis by using PCR-DGGE technique. The representative DGGE bands were chosen to be cloned, and sequenced, the phylogeny were constructed. The results showed that the bacterial communities were very different between the normal and diseased soil samples of C. chinensis, and the diversity index (H) of diseased soil samples were higher than that of normal soil samples. Sequencing analysis of representative cloned DGGE bands showed that the unculturable bacteria were the dominant groups, and bacteria belonged to genus Bacillus, Acidovorax, Acinetobacter, uncultured Kluyvera, and uncultured Comamonas were also existing, but the reported plant pathogenic bacteria were not found in the C. chinensis planting soil. The density and brightness of clone band d in diseased soil samples was higher than that in normal soil sample, and sequencing analysis showed that it belonged to genus Acidovorax. Obviously, during the process of C. chinensis growth, the rhizospheric bacteria population changed, and the quantity of bacteria belong Acidovorax increased, which probably resulted in the disease occurred during C. chinensis growth.
Bacteria ; classification ; genetics ; isolation & purification ; Biodiversity ; Coptis ; growth & development ; microbiology ; Denaturing Gradient Gel Electrophoresis ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Rhizosphere ; Soil Microbiology

OBJECTIVETo investigate the effect of human caudal-related homeobox 2 (Cdx2) gene expression on human gastric carcinoma cell line MGC-803.

METHODSpCMV-Cdx2-HA eukaryotic expression plasmid was constructed by using DNA recombinant method. The MGC-803 cells were divided into 4 groups: non-transfected, transfected with pCMV-HA, transfected with pCMV-GAPDH-HA, transfected with pCMV-Cdx2-HA. Cdx2 gene and its protein expression was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot techniques respectively. The effects of Cdx2 overexpression on the growth of MGC-803 cells in vitro was assessed by measuring MTT, flow cytometry and transmission electron microscopy analysis.

RESULTSIn MGC-803 cells transfected with pCMV-Cdx2-HA, cell proliferation was significantly suppressed, the cells was ultrastructurally destroyed, cell cycle progression was blocked in G0/G1 and apoptosis rate was higher (P<0.05) in comparison with non-transfected cells or cells transfected with empty vector.

CONCLUSIONIt indicated that Cdx2 gene can suppress the growth of gastric carcinoma and may save as a novel therapeutic target.

Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Genetic Vectors ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Plasmids ; genetics ; RNA, Messenger ; genetics ; Stomach Neoplasms ; metabolism ; pathology ; Transfection

Actins ; metabolism ; Animals ; Brain ; cytology ; metabolism ; Cell Culture Techniques ; methods ; Cells, Cultured ; Immunohistochemistry ; Pericytes ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor, Platelet-Derived Growth Factor beta ; metabolism ; Staining and Labeling

OBJECTIVETo clone a novel human testis-specific gene TDRG1.

METHODSA new contig of expression sequence tags (ESTs) Hs.180197 was identified from the testis libraries using digital differential display (DDD) to screen the novel human testis-specific gene. To validate the use of bioinformatics approaches in gene discovery, the ESTs Hs.180197, which was predicted to be testis specific, was chosen for further study. Reverse transcriptase-polymerase chain reaction (RT-PCR) was performed on different normal tissues to identify the expression of Hs.180197 in human testis. Using bioinformatics methods and IMAGE cloning of this contig, the full-length cDNA sequence of the noval human gene was cloned.

RESULTSThis novel gene was 1197 bp in length, located in chromosome 6p21.1-p21.2. The sequence of the open reading frame was 504-806 bp, as confirmed by RT-PCR and sequencing in human testis. The cDNA encodes a novel protein of 100 amino acids with a theoretical molecular weight of 10 000 and isoelectric point of 6.81. The sequence shares no significant homology with any known protein in the databases. Semi-quantitative RT-PCR analysis of multiple tissues further showed that the novel gene was expressed specifically in adult human testis. Considering a possible relation of this novel gene with the function of human testis, we named this new gene TDRG1 (testis development related gene 1, GenBank accession number: DQ168992).

CONCLUSIONDDD combined with laboratory validation is an efficient method for identifying new human functional genes.

Adult ; Cloning, Molecular ; DNA, Complementary ; genetics ; Databases, Genetic ; Gene Expression Regulation ; Homeodomain Proteins ; genetics ; Humans ; Male ; Molecular Sequence Data ; Organ Specificity ; Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment ; Testis ; metabolism

OBJECTIVETo explorer the effectiveness of enriched bone marrow stem cells technique for lumbar fusion.

METHODSWith the randomization and control principles, 2 graft materials [Enrichment bone marrow mesenchymal stem cells hybridized with beta-tri calcium phosphate (composite graft group), autologous iliac crest bone graft (autograft group)] were compared in posterior lumbar fusion procedures. 56 patients with degenerative disc disease, lumbar instability or spinal stenosis, were included. The volume of cells suspension in pre- and post-enrichment and the number of nucleated cells (NCs) were identified. The number of osteoprogenitor cells was estimated by counting the colony-forming units which express alkaline phosphatase (CFUs/ALP+). Then the efficiency of the enrichment was evaluated. Clinical follow-up with roentgenogram and Oswestry scale scores was performed for outcome evaluation.

RESULTS(249 +/- 31) ml bone marrow per patient from bilateral iliac crests was aspirated peri-operatively. About (43 +/- 11) ml enriched bone marrow was collected. The number of NCs was concentrated from (15.9 +/- 3.3) x 10(6)/ml to (44.1 +/- 10.8) x 10(6)/ml, CFUs/ALP+ was significantly increased from (118 +/- 86)/ml to(486 +/- 305)/ml. The follow-up was about (26.3 +/- 7.5) months. There was no significant differences in age, gender, disease and fusion segments between the two groups. The fusion rate was 93.3% and 96.2% for composite graft group and autograft group, respectively (chi2 = 0.2146, P = 0.6432). There was no difference in operation time between the two group (t = 0.5243, P = 0.6022), but blood loss in composite graft group was more than that in autograft group (t = 6.4664, P < 0.01). Cell salvage for auto-transfusion could transfuse back half of the blood loss during operation. No hematoma or chronic soreness in the bone marrow donor sites of composite graft group occurred, but a little exudation or moderate swelling in the wound happened in 4 cases which disappeared under medical treatment. Meanwhile, 15.4% patients had hematoma in the iliac bone donor site and 26.9% patients had chronic soreness, but no case had wound problem in autograft group. As for Oswestry scale scores, there was no significant difference between the two groups.

CONCLUSIONSThe enrichment technique of autologous bone marrow stem cells can greatly increase the concentration of MSCs. It is a rapid and safe method used peri-operatively. The composite material of enriched MSCs and porous beta-TCP is a good bone substitute in posterior spinal fusion.

Adult ; Aged ; Bone Substitutes ; Bone Transplantation ; methods ; Calcium Phosphates ; Female ; Follow-Up Studies ; Humans ; Ilium ; transplantation ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Spinal Fusion ; instrumentation ; methods ; Stem Cell Transplantation ; Transplantation, Autologous ; Treatment Outcome

OBJECTIVETo assess the effectiveness of tongjiang granule (TJG) on the patients with nonerosive reflux disease (NERD) of Gan-Wei incoordination syndrome, its impact on their quality of life, and its safety.

METHODA randomized, controlled, double-blinded, and double-dummy method was adopted in the trial. There were 120 NERD patients enrolled in the study and randomly divided into the experiment and control groups, each with 60 patients; drugs were distributed according to the drug number by patients' inclusion sequences. In the experiment group, patients were given TJG 10 g and mosapride citrate dummy 5 mg three times a day, and in the control group, patients were given mosapride citrate 5 mg and TJG dummy 10 g three times a day. The treatment courses of both groups were 4 weeks.

RESULTSAmong 120 included patients, 112 were screened for full analysis set (FAS), and 105 were screened per-protocol set (PPS). The results were as follows: (1) the improvement of total scores of symptom in the experiment group (0-4 week) were 15.93±7.88 scores by FAS and 16.22 ±7.75 scores by PPS, and they were 10.43±10.16 scores and 10.79±10.27 scores in the control group, respectively. The 95% CI of net scores improvement between the two groups were 2.10-8.90 scores and 1.92-8.94 scores in FAS and PPS; it was significantly better in the experiment group than that in the control group (P<0.05). (2) The improvement of scores of major symptom in the experiment group (0-4 week) were 10.68±5.35 by FAS and 10.89±5.29 by PPS and 7.40±7.41 and 7.60±7.46 in the control group, respectively. The 95% CI of net scores improvement in the two groups were 0.85-5.71 and 0.71-5.69 in FAS and PPS separately, and the improvement in the experiment group was significantly better than that in the control group (P<0.05). (3) The total effective rates were 86.0% and 61.8% in the experiment and the control group separately, and the Ridit analysis results showed that it was better in the experiment group (P<0.05). (4) The improvement quality of life in the domain of physical functioning and general health in the experiment group was better than that in the control group (P<0.05). (5) One case of experiment group caught a cold and recovered in six days without drug suspension. No adverse event was found in the other cases. There was no meaningful safety examination indices change in pretreatment and posttreatment periods in both groups.

CONCLUSIONTJG showed a definite effect on the treatment of NERD with Gan-Wei incoordination syndrome, and it could improve the quality of life of NERD patient without obvious toxic and side effects.

Case-Control Studies ; Double-Blind Method ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Gastroesophageal Reflux ; complications ; drug therapy ; Humans ; Male ; Middle Aged ; Quality of Life ; Syndrome ; Treatment Outcome

OBJECTIVETo clone UreB gene of Helicobacter pylori (H. pylori) isolated from children to pGEX-4T-1 expression plasmid, and do sequence analysis.

METHODSA pair of specific primer was designed according to H. pylori UreB gene in the GenBank. Using H. pylori strains isolated from children as a template, a UreB gene was obtained by PCR. After EcoR I and Not I digestion, the PCR production was linked with pGEX-4T-1 which was digested with the same enzymes. The recombinant plasmid was transformed into E.coli BL21 and identified by double enzyme digestion and sequence analysis. The sequence results were compared with the gene sequence in the GenBank.

RESULTSA UreB gene was successfully amplified from children's H. pylori strain GZCH1. It was 1710 bp in size. The objective band was identified by double enzyme digestion. DNA sequence showed that UreB was in the correct open reading frame. The sequence comparison analysis showed that DNA and amino acid sequence identities of UreB gene with other strains were 98%. The sequence of UreB of H. pylori strain GZCH1 was submitted to GenBank (accession number:FJ455126).

CONCLUSIONSUreB of H. pylori strain GZCH1 is successfully cloned to pGEX-4T-1, which provides a basis for research of oral H. pylori vaccine.

Amino Acid Sequence ; Bacterial Vaccines ; immunology ; Child ; Cloning, Molecular ; Helicobacter pylori ; enzymology ; immunology ; Humans ; Male ; Molecular Sequence Data ; Urease ; chemistry ; genetics ; immunology