Objective To detect molecular characterization of the promoter and 5′UTR region of ATP7B gene in Chinese children with Wilson disease (WD) and explore the distribution of polymorphisms and mutations in different ethnicity.Methods One hundred and ten patients with WD and 90 healthy children were enrolled into the study and analyzed by polymerase chain reaction-single strand configuration polymorphism (PCR-SSCP) and DNA sequence analysis.Results 1.Five polymorphisms were identified as follows, -1294T→G,-105C→G,-116C→T ,-132delGCCGC and -75A→C(transcription start site as +1). The former three ones had never been reported before. The fourth one had not been reported either in China. 2.The polymorphism -132delGCCGC and -75A→C both exhibited with allelic frequency at above 70%, which was much higher than other races. The -132delGCCGC polymorphism shared almost complete linkage disequilibrium with the -75A→C polymorphism (in 98% patients) and their regularity was 96.9%.3. Almost all the polymorphisms distributed in flanking sequence of EXON 1 in Chinese. Race and geological distribution may be dominant factors of the variability of promoter and 5′UTR region of ATP7B gene.Conclusions Three novel polymorphisms and a linkage disequilibrium with the -132delGCCGC and -75A→C were identified in Chinese with WD. It also suggests that the mutation in the promoter of ATP7B is uncommon in Chinese patients.

OBJECTIVETo explore the myelo-protection effect of mdr1 transfected cord blood cells (CBMNCs) graft against high-dose homoharringtonine leukemia-bearing severe combined immunodeficient (SCID) mice model.

METHODSMultidrug resistant (mdr1)gene was transferred into CBMNCs by a retrovirus vector, containing full-length cDNA of human mdr1 gene. CBMNCs and high-titer retrovirus supernatant were cocultured with cytokine combinations for 5 - 6 days. The SCID mouse models bearing human HL-60 cell leukemia were divided into three groups. Group A received tail vein injection of 2 x 10(6) mdr1 gene transduced CBMNCs at day 1 and 3, groups B and C 2 x 10(6) un-transduced CBMNCs and same volume of normal saline, respectively. The 3 groups of the mouse model were treated with weekly escalated doses of homoharringtonine. The peripheral white blood cell (WBC) counts, the human leukemia cells percentage in peripheral blood, the histological findings of main organs were assayed. The CD33 positive HL-60 cells in bone marrow were determined by flow cytometry. The function and expression of mdr1 gene were examined by PCR, immunochemistry (IC) and DNR extrusion test in vivo.

RESULTS(1) mdr1 gene was transferred into CBMNCs successfully and the transfection frequency was 30%. (2) Leukemia SCID mice were xenotransplanted with mdr1-transfected BMMNCs by a programmed procedure and could be used as a valuable model for in vivo evaluating myelo-protection effects. (3) The transfected mice could tolerate homoharringtonine 5 approximately 6 folds higher than conventional dose and kept peripheral WBC count at a mean of 3 x 10(9)/L, with the peripheral human myeloid leukemia cells percentage decreasing to less than 5%. Histological examination showed that there was no leukemia infiltration in the main organs, the CD33 positive HL-60 cells in bone marrow were less than 5%. (4) The repopulation frequency of the transfected CBMNs in marrow were 9.13%. DNR extrusion test confirmed that the P-gp product maintained its biological function in the marrow.

CONCLUSIONmdr1 transferred-human CBMNC can xenotransplanted and repopulated in leukemia-bearing SCID mouse and are protected from chemotherapy-induced myelosuppression.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; adverse effects ; therapeutic use ; Cord Blood Stem Cell Transplantation ; methods ; Female ; Fetal Blood ; cytology ; Genetic Vectors ; HL-60 Cells ; Harringtonines ; administration & dosage ; adverse effects ; therapeutic use ; Humans ; Leukemia, Promyelocytic, Acute ; drug therapy ; pathology ; surgery ; Leukocytes, Mononuclear ; cytology ; metabolism ; transplantation ; Male ; Mice ; Mice, SCID ; Random Allocation ; Retroviridae ; genetics ; Transfection ; Treatment Outcome ; Xenograft Model Antitumor Assays

OBJECTIVETo observe the effect of total coptis alkaloids (TCA) on beta -amyloid peptide (A beta 25-35) induced learning and memory dysfunction in rats, and to explore its mechanism.

METHODSForty male Wistar rats were randomly divided into four groups: the control group, the model group, the TCA low dose (60 mg/kg) group and the TCA high dose (120 mg/kg) group, 10 in each. A beta 25-35 (5microl, 2 microg/microl) was injected into bilateral hippocampi of each rat to induce learning and memory dysfunction. TCA were administered through intragavage for consecutive 15 days. Morris Water Maze test was used to assess the impairment of learning and memory; concentration of malondialdehyde (MDA) in cerebral cortex was determined by thiobarbituric acid reactive substance to indicate the level of lipid peroxidation in brain tissues; activity of manganese-superoxide dismutase (Mn-SOD) in cerebral cortex was determined by xanthine-oxidase to indicate the activity of the enzyme; and NF- kappa B protein expression in cerebral cortex was measured by SP immunohistochemistry.

RESULTS(1) Morris Water Maze test showed that, during the 4 consecutive days of acquisition trials, the rats in the model group took longer latency and searching distance than those in the control group (P<0.01), which could be shortened by high dose TCA (P<0.05); during the spatial probe trial on the fifth day, the rats in the model group took shorter searching time and distance on the previous flat area than those in the control group (P<0.01), which could be prolonged after TCA treatment (for low dose group, P<0.05; for high dose group, P<0.01). (2) Analysis of cerebral cortical tissues showed that, compared with the control group, MDA level got significantly increased and Mn-SOD activity decreased in the model group (both P<0.01). After having been treated with TCA, the MDA level got significantly decreased (P<0.05 and P<0.01 respectively for low and high dose group), while relative increase of Mn-SOD activity only appeared in high dose group (P<0.05). (3) Immunohistochemistry analysis showed the protein expression of NF- kappa B got significantly increased after modeling, while high dose TCA can significantly inhibit it.

CONCLUSIONTCA could improve A beta 25-35 induced dysfunction of learning and memory in rats, and its protective mechanism is associated with its actions in decreasing MDA level, increasing Mn-SOD activity and inhibiting the expression of NF-kappa B in cerebral cortex.

Alkaloids ; administration & dosage ; pharmacology ; Amyloid beta-Peptides ; administration & dosage ; Animals ; Cerebral Cortex ; metabolism ; Coptis ; chemistry ; Dose-Response Relationship, Drug ; Hippocampus ; drug effects ; Injections ; Learning Disorders ; chemically induced ; psychology ; Male ; Malondialdehyde ; metabolism ; Maze Learning ; drug effects ; Memory ; drug effects ; Memory Disorders ; chemically induced ; psychology ; NF-kappa B ; metabolism ; Peptide Fragments ; administration & dosage ; Rats ; Rats, Wistar ; Reaction Time ; drug effects ; Superoxide Dismutase ; metabolism ; Swimming

OBJECTIVETo investigate the distribution of burn pathogens and their antibiotic resistance in a burn unit, so as to provide reference for clinical practice.

METHODSThree hundred and forty-eight burn patients hospitalized in our department were enrolled in this study. The pathogens isolated from the wounds, blood, venous catheter, sputum, urine, purulent discharge of wounds in these patients, and their antibiotic resistance were surveyed by retrospective analysis from Jan, 2001 to Dec, 2006.

RESULTSTotal-ly 464 strains were isolated, among which Gram negative (G-) bacilli accounted for 52.6%, Gram positive microorganisms (G+) accounted for 40.5%, and fungi accounted for 6.9%. The main pathogens were Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter species and Escherichia coli, among which Staphylococcus aureus (MRSA) was predominant (93.5%). MRSA was 100% resistant to levofloxacin, penicillium, oxacillin, and it was also resistant to other antibiotics except Vancomycin. The resistance rate of Pseudomonas aeruginosa to Cefoperazone/Sulbactam, Imipenem and cefepime were 15.8%, 36.8%, 33.3%, respectively.

CONCLUSIONStaphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter species and Escherichia coli were predominant in the burn unit,among them Staphylococcus aureus and Acinetobacter were more resistant to antibiotics.

Acinetobacter baumannii ; drug effects ; isolation & purification ; Burn Units ; Burns ; microbiology ; Cross Infection ; microbiology ; Drug Resistance, Bacterial ; Escherichia coli ; drug effects ; isolation & purification ; Humans ; Pseudomonas aeruginosa ; drug effects ; isolation & purification ; Retrospective Studies ; Staphylococcus aureus ; drug effects ; isolation & purification

Current studies have demonstrated that SLC38A1 proteins play a causal role in neoplastic cell transformation.The twofold aim of this study was to provide insight into whether a variance in the expression of SLC38A1 exists between human colorectal cancer and healthy human tissues and to determine how silencing or overexpressing the SLC38A1 gene could affect the proliferation,viability and migration of colorectal cancer cells.Immunohistochemical staining was used to analyze the expression of SLC38A1 in colorectal cancer tissues and adjacent normal mucosa in 77 patients who underwent surgical resection.The expression of SLC38A1 in colorectal cancer tissues and cell lines was detected using RT-PCR and Western blotting.Two colorectal cancer cell lines SW480 and HCT116 were used to examine whether silencing SLC38A1 with siRNA and overexpressing SLC38A1 with shRNA could affect cell viability and migration.As a result,the SLC38A1 protein was very low or undetectable in the normal colon mucosa.In contrast,strong staining of SLC38A1 protein was found in the cytoplasm in 79.2％ colorectal cancer samples.More pronounced SLC38A1 expression in colorectal cancer tissues was significantly associated with tumor node metastasis (TNM) stage.Inhibition of SLC38A1 reduced tumour growth and suppressed proliferation and migration of SW480 cells.In contrast,overexpression of SLC38A1 had the opposite effects on HCT116 cells.S LC38A1 is overexpressed in colorectal cancer,which suggests that it is associated with tumour progression.These results encourage the exploration of SLC38A1 as a target for intervention in colorectal cancer.

OBJECTIVETo investigate the effects of top pruning on fruiting characters of Platycodon grandiflorum, and find the suitable stage, in which seed growth and development furtherly.

METHODOne-year old seedlings were chosen and planted in field. Plant height, branching number, fruit number per plant, 1000 grains weight were measured during growth and development period, respectively.

RESULTThe treatment of top pruning postponed in turn the flowering date, lowered the plant heights and the fruit number per plant, increased the branching number and influenced significantly on 1000 grains weights.

CONCLUSIONThe suitable stage of top pruning for producing seeds was from June 20th to July 5th.

Crops, Agricultural ; growth & development ; Fruit ; growth & development ; Platycodon ; growth & development

OBJECTIVETo investigate the clinical response, pathological complete response (pCR), tumor resection rate and safety of neoadjuvant chemotherapy with docetaxel and epirubicin (ET) for locally advanced breast cancer (LABC).

METHODSFrom March to December 2001, 40 women with LABC, aged from 28-67 (medium 48) years were alloted. Twenty patients had clinical stage IIIa disease, 15 had stage IIIb disease and 5 stage IV patients who had ipsilateral sura-clavicular metastasis. The dose was: epirubicin (E) 60 mg/m2, docetaxel (T) 75 mg/m2 every 3 weeks, with G-CSF given preventively. After 2 cycles of ET, a pilot clinical response evaluation was performed by investigators for each patient to decide if she should receive another 1-2 cycles of ET before surgery or radiation therapy.

RESULTSThirty-eight patients received 2-3 cycles of ET regimen. The pCR, clinical complete response (cCR) and clinical partial response (cPR) rates were 15.0%, 20.0% and 52.5%, respectively. Tumor resection rate in this group was 92.5%. Incidence of III/IV Grade neutropenia was 8.4%/14.0% of cycles, and 3 patients suffered from neutropenia with fever. Non-hematological adverse events were alopecia, nausea, vomiting, fluid retention, myalgia, arthralgia and nail disorders, which were mild to moderate.

CONCLUSIONNeo-adjuvant chemotherapy with a combination of docetaxel and epirubicin is effective and well tolerated by women with locally advanced breast cancer.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; adverse effects ; therapeutic use ; Breast Neoplasms ; drug therapy ; surgery ; Carcinoma, Ductal, Breast ; drug therapy ; surgery ; Chemotherapy, Adjuvant ; Drug Administration Schedule ; Epirubicin ; administration & dosage ; adverse effects ; Female ; Humans ; Male ; Middle Aged ; Neutropenia ; chemically induced ; Paclitaxel ; administration & dosage ; adverse effects ; Remission Induction

Objective To learn the prevalence, distribution and risk factors of mild cognitive impairment (MCI) among elderly in Zhoushan City, and to provide the reference for MCI control. Methods A total of 1801 elderly people aged 60 to 79 years old from six districts of Zhoushan City were sampled by method of stratified random sampling. After self-evaluated with Ascertain Dementia 8 (AD8) and screened with Screening Scale for Mild Cognitive Impairment (sMCI), the diagnosis by specialists was conducted for that positive to AD8 and sMCI. Results of 1801 respondents, 873 (48.47％) people were male, and the other 928 (51.53％) people were female; 38.65％ of the people selected aged 60-<65; 90.28％ had a primary school education or were illiterate; 78.51％ had legitimate and healthy wives. A total of 122 elderly people were diagnosed with MCI, and the prevalence of MCI was 6.77％. Multivariate logistic regression analysis showed that male elderly people (OR=0.53, 95％CI:0.28-1.00) were less likely to develop MCI compared to the female, and the illiterate (OR=2.09, 95％CI: 1.16-3.77) were more likely to develop MCI compared to the educated . Conclusion The prevalence of MCI among the elderly in Zhoushan was 6.77％; the female and the illiterate were more likely to develop MCI.

The sialyl Lewis X (SLex) antigen encoded by the FUT7 gene is the ligand of endotheliam-selectin (E-selectin).The combination of SLex antigen and E-selectin represents an important way for malignant tumor metastasis.In the present study,the effect of the SLeX-binding DNA aptamer on the adhesion and metastasis of hepatocellular carcinoma HepG2 cells in vitro was investigated.Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence staining were conducted to detect the expression of FUT7 at both transcriptional and translational levels.The SLex expression in HepG2 cells treated with different concentrations of SLeX-binding DNA aptamer was detected by flow cytometry.Besides,the adhesion,migration,and invasion of HepG2 cells were measured by cell adhesion assay,and the Transwell migration and invasion assay.The results showed that the FUT7 expression was up-regulated at both mRNA and protein levels in HepG2 cells.SLeX-binding DNA aptamer could significantly decrease the expression of SLex in HepG2 cells.The cell adhesion assay revealed that the SLeX-binding DNA aptamer could effectively inhibit the interactions between E-selectin and SLex in the HepG2 cells.Additionally,SLeX-binding DNA aptamers at 20 nmol/L were found to have the similar effect to the monoclonal antibody CSLEX-1.The Transwell migration and invasion assay revealed that the number of penetrating cells on the down-side of Transwell membrane was significantly less in cells treated with 5,10,20 nmol/L SLeX-binding DNA aptamer than those in the negative control group (P＜0.01).Our study demonstrated that the SLeX-binding DNA aptamer could significantly inhibit the in vitro adhesion,migration,and invasion of HepG2 cells,suggesting that the SLeX-binding DNA aptamer may be used as a potential molecular targeted drug against metastatic hepatocellular carcinoma.

Objective To investigate the influence of surgery with the guideline of minimally invasive concept in prognosis of patients with hypertensive basal ganglia hematomas. Methods Fifty-seven patients with hypertensive intra-cerebral hemorrhage were randomly divided into 2 groups:Group A (admitted to our hospital from January 2007 to December 2008 and performed surgery under the condition that the content of hematoma reached the level for surgery,n=26) and Group B (admitted to our hospital from January 2009 to June 2011 and received surgery with the guideline of minimally invasive concept once noting the tendency ofexpanded hematoma,n=31).We evaluated the influence of surgery (total removal of the hematoma and proper stopping the bleeding) according to the condition that tendency of expanded hematoma appeared and with the guideline of minimally invasive concept in the prognosis of these patients. Results No significant differences in consciousness classification and hematoma volume before surgery were noted between the 2 groups (P＞0.05).Responsible vessels were noted in 15 patients from Group A and 27 patients from Group B, and significant difference was noted between these 2 groups (P＜0.05).The hematoma clearance rate was 75％ in Group A,and re-bleeding was noted in 4 patients (15.4％) after the surgery; while that was higher than 90％ in Group B, and re-bleeding was only noted in 2 patients (6.5％) whose responsible vessels could not be found.The good recovery rate in Group A was 46.2％ 3 months after surgery, while that in Group B was 74.2％, which indicated that the effect in group B was obviously better than that in group A (P＜0.05). Conclusion Tendency of expanded hematoma should be paid attention in patients with hypertensive basal ganglia cerebral hemorrhage; it is important to quickly identify the cases showing clear indications for surgery and to perform the procedures at the earliest time; the procedures, including completely removal of the hematoma and properly stopping the stanch bleeding under direct vision with the guideline of minimally invasive concept can improve the recovery fiom hypertensive basal ganglia cerebral hemorrhage.