Liquid chip technology have been licensed to be used in clinic because of its advantage of high-throughput, high-sensitivity, good signal to noise ratio, reaction in liquid phase, convenient operation and short time consuming, etc. The optimization of a liquid chip system for the detection of serum biomarkers of colorectal tumour and initial application in the detection of CEA were studied. The optimized reaction conditions of liquid chip were determined through orthogonal design after it was prepared. The results showed that the consuming reaction time of the coated antibody and the antigen was 1hour. The microspheres, biotinylated detecion antibody and the consuming complexes and avidin-PE time of the microspheres and the biotinylated tested antibody was 1hour, 1hour and 15minutes respectively.the consuming time of the complexes and avidin-PE was fifteen minutes, The optimized dilution of the biotinylated tested detection antibody was 1∶300 and the optimized concentration of avidin-PE was 12?g/ml. Totally 55 clinical samples were detected by the liquid chip and by Enzyme-Linked Immunosorbent Assay (ELISA) simultaneously and the results of the two methods were compared. The results of the two methods showed good correlation between positive and negative samples but the detection limits and the dynamic ranges of the liquid chip method were more sensitive and wider than those of the ELISA. The multiple tumour biomarkers may be detected simultaneously and the time of clinical test and manpower requirements were reduced by the liquid chip method.

OBJECTIVETo examine the protective effect of Ginkgo biloba leaf extract (GbE) on learning and memory deficit induced by aluminum chloride (AlCl(3)), and explore its mechanisms.

METHODSThe rat models with learning and memory deficit were induced by administering via gastrogavage and drinking of AlCl(3) solution. And the model rats were treated with GbE at the dose of 50, 100, 200 mg/kg every day for 2 months accompanied with drinking of AlCl(3) solution, respectively. Their abilities of spatial learning and memory were tested by Morris water maze, and the acetylcholinesterase (AChE) activity in serum was assayed with chemical method, the AChE expression in hippocampus was observed by immunohistochemistry assay, and then quantitative analysis was done by BI 2000 image analysis system.

RESULTSLearning and memory deficit of rats could be induced by AlCl(3) solution (P < 0.01), and AChE expressions in rats hippocampus were increased (P < 0.01); GbE ameliorated learning and memory deficit and reduced AChE expression in rats hippocampus in a dose-dependent manner, while GbE significantly increased serum AChE activity at the dose of 200 mg/kg each day (P < 0.05).

CONCLUSIONGbE can ameliorate learning and memory deficit induced by AlCl(3), which may be due to its inhibition of the AChE expression in hippocampus.

Acetylcholinesterase ; metabolism ; Aluminum Compounds ; toxicity ; Animals ; Chlorides ; toxicity ; Dose-Response Relationship, Drug ; Ginkgo biloba ; Hippocampus ; enzymology ; Immunohistochemistry ; Male ; Maze Learning ; drug effects ; Memory Disorders ; chemically induced ; prevention & control ; Neuroprotective Agents ; therapeutic use ; Phytotherapy ; Plant Extracts ; therapeutic use ; Plant Leaves ; Plant Structures ; Rats ; Rats, Wistar ; Reaction Time

Acute Disease ; Anemia, Refractory ; etiology ; therapy ; Child ; Child, Preschool ; Cord Blood Stem Cell Transplantation ; methods ; Female ; Histocompatibility Testing ; Humans ; Leukemia ; complications ; pathology ; therapy ; Male ; Treatment Outcome

This research established an HPLC method for determination of six C-Glycoside flavones of warer-soluble total flavonoids from Isodon lophanthoides var. gerardianus (Benth.) H. Hara, and studied the antitumor activity of the warer-soluble total flavonoids. The HPLC system consisted of Kromasil 100-5 C18 (4.6 mm x 250 mm, 5 microm) column and a solution system of methanol, acetonitrile and 0.5% formic acid gradient elution at a flow rate of 0. 8 mL x min(-1) and the wavelength of detector was at 334 nm. The column temperature was 25 degrees C. The antitumor activity of water-soluble flavonoids was assayed using HepG2 cell as the tested cell. The linear ranges of vicenin II, vicenin III, isoschaftoside, schaftoside, vitexin, 6, 8-di-C-a-L-arabinosylapigenin were 0.25-2.53, 0.12-1.20, 0.37-3.69, 0.16-1.63, 0.19-1.92, 0.14-1.42 microg, respectively. The average recoveries (n = 6) were 99.6% (RSD 0.87%), 100.2% (RSD 2.0%), 99.6% (RSD 1.8%), 97.9% (RSD 1.5%), 98.8% (RSD 1.2%), 98.6% (RSD 1.2%), respectively. After exposure in 24, 48, 72 h, the total flavonoids showed inhibitory effect on the proliferation of HepG2 cells with IC50 as the evaluation index, the IC50 values of 1.89, 1.71, 1.51 g x L(-1), respectively. The method is quick, simple and accurate with good re- producibility, and can be used for determination of vicenin II, vicenin III, isoschaftoside, schaftoside, vitexin, 6, 8-di-C-a-L-arabino- sylapigenin in the warer-soluble total flavonoids from L lophanthoides var. gerardianus. The warer-soluble total flavonoids from L lophanthoides have inhibitory effect on the proliferation of HepG2 cells.
Antineoplastic Agents, Phytogenic ; analysis ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Flavones ; analysis ; pharmacology ; Humans ; Isodon ; chemistry ; Monosaccharides ; analysis ; pharmacology

OBJECTIVETo explore the mechanism of rehabilitation after middle cerebral artery occlusion (MCAO).

METHODSMCAO model was reproduced with two-kidney, two clip renovascular hypertensive rats stroke-prone (RHRSP), which were divided into two groups, the treated group (treated with electric stimulus) and the control group (untreated model) randomly. The rehabilitation of rats was evaluated by balance beam walking test. The ultrastructural changes of neurons and astrocytes, expressions of glial fibrillary acidic protein (GFAP)-positive cells, neurofilament (NF) protein, and cerebral capillary dilatation M-associated protein-2 (MAP2), as well as the neurons apoptosis and the number of dilatation of cerebral capillary in the margin of infarcted area were observed by the end of 1st, 3rd, 6th and 9th week after modeling.

RESULTSThe motor function of paralysed limbs recovered better in the treated group than that in the control group by the end of 3-9th week after MCAO, the expression of GFAP-positive cells in astrocytes and NF, MAP2 in neurons as well as the number of cerebral capillary dilatation at the margin of infarcted area were higher than those in the control group (P < 0.05).

CONCLUSIONElectric stimulation treatment could improve the recovery of motor function of paralyzed limbs. It might be due to the effect of electric stimulus in increasing astrocytes proliferation, reinforcing activity of neurons and evoking the dilatation of cerebral capillary.

Animals ; Apoptosis ; Astrocytes ; ultrastructure ; Electric Stimulation Therapy ; Glial Fibrillary Acidic Protein ; metabolism ; Infarction, Middle Cerebral Artery ; pathology ; physiopathology ; rehabilitation ; Male ; Neurons ; ultrastructure ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To develop a multiplex allele-specific PCR (AS-PCR) assay with three-color fluorescence labeling for mitochondrial DNA (mtDNA) SNP typing. METHODS: Based on the principle of AS-PCR, the primer sets were designed for 20 SNP located on the coding region of mtDNA and divided into 2 groups labeled with FAM and HEX fluorescence, respectively. A primer set included two forward (reverse) allelic specific primers with different sizes and a generic reverse (forward) primer. Blood samples from 200 unrelated individuals were analyzed by AS-PCR and capillary electrophoresis. Three random samples at least for each SNP site were examined and verified by direct sequencing. The haplotype frequency was investigated. RESULTS: Distinct electropherograms of 200 blood samples were obtained successfully. The typing results of direct sequencing were identical to those obtained from AS-PCR. The minimum detectable DNA concentration was 0.2 pg under the system of 10 microL. The sensitivity of the DNA concentrations ranged from 0.5 to 5 pg. The 200 individuals were assigned into 15 haplotype, and the haplotype diversity was 0.906 0. CONCLUSION AS-PCR is a simple, rapid and efficient method for mtDNA SNP typing, and can be applied to forensic practice.
Alleles ; DNA ; DNA Primers ; DNA, Mitochondrial/analysis* ; Electrophoresis, Capillary ; Haplotypes ; Humans ; Mitochondria ; Polymerase Chain Reaction/methods* ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA

Blood Cell Count ; Bone Marrow Cells ; metabolism ; Child ; Cord Blood Stem Cell Transplantation ; Cytokines ; therapeutic use ; Histocompatibility Testing ; Humans ; Male ; Myelodysplastic Syndromes ; blood ; therapy ; Treatment Outcome

OBJECTIVETo explore operative complications of photoselective vaporization of prostate (120 W) for treatment of benign prostatic hyperplasia (BPH).

METHODSThe clinical data of 186 cases who underwent photoselective vaporization of prostate (120 W) for the treatment of BPH from May 2010 to April 2012, was statistically analyzed.

RESULTSThe operative time ranged from 7 to 147 minutes, and the average time was (37.7 ± 21.5) minutes. No patient accepted intraoperative blood transfusion, and occurred transurethral resection syndrome or capsular perforation. The time of postoperative indwelling catheter ranged from 1 to 11 days, and average time was (4.3 ± 2.2) days. Surgical outcome was satisfactory. Early postoperative complications included bladder spasm (3 cases), transient dysuria (19 cases), urinary tractirritation (94 cases), secondary hemorrhage (26 cases), transient urge incontinence (19 cases), all cases were relieved after treatment. Long-term complications, including recurrence (1 case), bladder neck stenosis (2 cases) and urethral stricture (2 cases), who had required reoperation. Postoperative patients with international prostate symptom score (29.4 ± 3.4), maximum urinary flow rate ((6.0 ± 1.6) ml/s) and residual urine ((167 ± 150) ml) had improved (t = -76.0 - 61.4, P < 0.01).

CONCLUSIONSWith less invasive, less bleeding and rapid postoperative recovery, photoselective vaporization of prostate (120 W) is a safe and effective minimally invasive treatment techniques for BPH. But there is still some complications after surgery and proper handling is required.

Aged ; Aged, 80 and over ; Humans ; Lasers, Solid-State ; Male ; Middle Aged ; Postoperative Complications ; epidemiology ; Prostate ; surgery ; Prostatic Hyperplasia ; surgery

OBJECTIVETo investigate the incidence of different types of brain injury in preterm infants and their influencing factors.

METHODSThe clinical data and head magnetic resonance imaging (MRI) findings of 239 preterm infants were collected, and the influence of antepartum, intrapartum, and postpartum factors on brain injury in preterm infants was analyzed.

RESULTSThe incidence rate of brain injury in preterm infants was 25.5%; among these infants, 10.5% had hemorrhagic brain injury, 10.5% had ischemic brain injury, and 4.6% and hemorrhagic and ischemic brain injury. The infants with a lower gestational age had higher incidence rates of hemorrhagic brain injury and overall brain injury (P<0.01). The incidence rates of ischemic brain injury and hemorrhagic and ischemic brain injury were not correlated with gestational age (P>0.05). The incidence rates of hemorrhagic, ischemic, and overall brain injury were not correlated with birth weight (P>0.05). Multiparity (OR=0.292, 95%CI 0.088-0.972) and cesarean section (OR=0.075, 95%CI 0.015-0.368) were protective factors against brain injury in infants with a gestational age of <34 weeks; cesarean section (OR=0.296, 95%CI 0.131-0.672) was the protective factor against brain injury in infants with a gestational age of ≤34 weeks, and severe infection (OR=8.176, 95%CI 1.202-55.617) was the risk factor.

CONCLUSIONSIn order to prevent or reduce the occurrence of brain injury in preterm infants. the gestational age of preterm infants should be prolonged as much as possible and the indications for cesarean section should be grasped. Infections should be prevented and if occurring should be treated actively and effectively.

Birth Weight ; Brain Injuries ; epidemiology ; etiology ; Female ; Gestational Age ; Humans ; Incidence ; Infant, Newborn ; Infant, Premature ; Male ; Retrospective Studies ; Risk Factors

Objective To clone and sequence the cysteine protease inhibitor gene of periodic Brugia malayi(BmCPI) and predict B-cell epitopes in amino acide sequence of BmCPI in order to provide basis for further study the expression of BmCPI and its function. Methods Total RNA was extracted from periodic Brugia malayi.A couple of specific primers were designed on the basis of known sequences of cysteine protease inhibitor gene from BmCPI. The desired gene was amplified by PCR technique from cDNA. The PCR products were purified and cloned into plasmid pGEM-T by T-A cloning method, transformed into Escherichia coli(E, coli) strain DH5α. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification. Five parameters and methods were used to predict B-cell epitopes in amino acide sequence of BmCPI. Results For RT-PCR, a specific band of around 621 bp was amplified. The same band was obtained by double restriction of recombinant plasmids or PCR using recombinant plasmid as template. The result of DNA sequencing showed that BmCPI shares 99% nucleotide sequence identity with that of published sequence. It showed that B-cell epitopes were probably at or adjacent to 23 - 32, 50 - 79 and 117 - 126 in its amino acide sequence. Conclusions pGEM-BmCPI is successfully constructed and sequenced, anticipated objective is reached and conditions is provided for further study of BmCPI expression and its function.