Objective To analysis the genomic sequence of a novel human leukocyte antigen (HLA)-B*3818 allele.Methods Full length genomic sequence of an unknown HLA-B allele was cloned,followed by bi-directional sequencing and the specificity of the antigen coded by this novel allele was defined by microcytotoxicity assay.The frequency and haplotype of this novel allele was acquired by population census and parentage analysis.Results The full length genomic sequence of this novel HLA-B*3818 allele with accession number FJ561482 differs from HLA-B*380201 by two nucleotide changes in exon 4 and intron 5,respectively.One change is located at nt 660 in exon 4 where C→A alternation,which results in an amino acid substitution from Asp(GAC)to Glu(GAA)at codon 196.This alternation is a new single nucleotide polymorphism compared with all other HLA-B alleles.Another is located at genomic position 2133 in intron 5(A→C).Except for this substitution,the intron sequences of HLA-B*3818 allele are identical to those of other HLA-B*38 alleles including HLA-B*380101,B*380201 and B*3814.The serological specificity of HLA-B*3818 is B38 and the frequency of this new allele is less than 0.000 5 in Chinese Han population.The parentage analysis showed the haplotype of novel allele is A*030101-Cw*010201-B*3818-DRB1*1312-DOB1*060101.Conclusion The simultaneous mutations in exon and intron were found in the Hovel HLA-B*3818 allele,and so it can present more sequence information for studies and applications associated with HIA genes by analyzing the genomic sequences of novel HLA alleles.

Objective To improve the knowledge and diagnostic ability of imagiology on neonatal respiratory distress syndrome (NRDS) computed radiograph(CR).Methods The doubtful patients were done to photographs bedside using the high resolution imaging plate, 50 cases of newborn with NRDS were selected whose clinical diagnosed clearly and had been treated and had the complete CR image documents.The CR change and clinical characteristics were observed dynamically.Results Nine of 50 cases were combined with aspirated pneumonia,8 cases with infective pneumonia,3 cases with intra-alveolar hemorrage,and 2 cases with pneumothorax.Accoding to X-ray manifestations,all cases were divided into four stages:Ⅰ stage(n=5), Ⅱ stage(n=20),Ⅲ stage(n=22),Ⅳstage(n=3).Typical CR signs included:the pulmonary lucency decreasd,wide-ranging net and grain shadowes of high density, and in companing with a lot of air brunchus sing.Conclusions Computed radiography is the most important imaging method in diagnosis of NRDS bedside ,and shall be improved the ability of diagnosis and differential of NRDS combined with the clinic.

Objective We examined the Grp78, ERK1/2 and phospho-ERK1/2 expressions in hepatocellular carcinoma(HCC) tissue samples in vitro, we interfered the expression of Grp78 in SMMC-7721 cells to explore whether Grp78 is involved in ERK1/2 signal pathway. Methods The Grp78, ERK1/2 and phospho-ERK1/2 expressions were detected by immunohistochemistry and confirmed by Western blotting in 47 HCC tissue samples. The Grp78 expression in SMMC-7721 cells was interfered by plasmid transfection and siRNA, ERK1/2 phosphorylation and expression were determined by Western blotting. Results The Grp78 expression was significantly correlated with ERK1/2 and phospho-ERK1/2 in HCC tissue samples. Overexpression of Grp78 promoted ERK1/2 phosphorylation in SMMC-7721 cells and the increased ERK1/2 phosphorylation was inhibited by Grp78 knockdown. Conclusion Grp78 is involved in the regulation of ERK1/2 signal pathway and might be a potential target for the comprehensive therapy of HCC.

OBJECTIVETo compare the ability of osthole (OST) and genistein (GEN) in enhancing bone peak bone mass of rats to prevent osteoporosis.

METHODSThirty-six female one-month-old SD rats of (125 +/- 3) g body weight were randomly divided into three groups, 12 rats in each group, one group was orally administered osthole at 9 mg x kg(-1) d(-1), one group was given genistein at 10 mg x kg(-1) d(-1) and another was given equal quantity of distilled water as the control. The body weight was monitored weekly and the bone mineral density (BMD) of total body was measured every month. All rats were sacrificed after three months, the femoral bone mineral density, the serum levels of osteocalcin (OC) and anti-tartaric acid phosphatase 5b (TRACP 5b) were measured by Elisa. The bone microarchitectures were analyzed with micro-CT and the bone biomechanics properties were tested with universal material machine.

RESULTSNo significant differences were observed between O-treated or GEN group and the control for the food-intake and body weight during three months. However, the rats treated with OST had significant higher BMD for both total body and femur than the control and GEN group. The O-treated rats also had higher level of serum OC and lower level of TRACP 5b. Besides, they owned bigger bone volume/tissue volume, trabecular thickness, trabecular number but smaller trabecular spacing. In the three point bending tests of femurs,they were found to have larger maximum load, the young's modulus and structural model index (SMI).

CONCLUSIONOrally administered osthole could efficiently increase the peak bone mass of rats,which provide new ideas for preventing osteoporosis.

Acid Phosphatase ; blood ; Animals ; Body Weight ; drug effects ; Bone Density ; drug effects ; Coumarins ; pharmacology ; Female ; Femur ; diagnostic imaging ; drug effects ; pathology ; Genistein ; pharmacology ; Isoenzymes ; blood ; Osteocalcin ; blood ; Radiography ; Rats ; Rats, Sprague-Dawley ; Tartrate-Resistant Acid Phosphatase

Objective To explore the present condition of endemic arsenism, the implementation of control measures and the effect of the monitored county (Tongyu County) and the monitoring spot (Baiyintuhai Village) in 2006 and 2007. Methods According to the National Survey Scheme of Endemic Arsenism, the progress of anti-arsenic water in Tongyu, and the management and running of all engineering projects and the arsenic content in water were surveyed. The patients with endemic arsenism in Tongyu were generally surveyed. The arsenic content of the improved drinking water in Tongyu and the arsenic in urine of children aged 8-12 and adults over 18 years of age were determinted. The causes of resident death in the monitoring spot from the year of 2006 and 2007 were investigated. Arsenic content of drinking water and the urine of local residents was examined with "Model AFS-930 Double-Channel Atomic Fluorescence Spectrometer". Results There were 30 endemic arsenism areas, 157 areas with high arsenic content and all population of 57 576 in Tongyu. Six areas had improved water till 2006, where 20.0% of water had improved, and six water-improving projects were all running normally, benefiting a population of 1670. Eight high arsenic areas changed water, in a rate of 5.09%. Eight water-improving projects were functioning well, benefiting a population of 4350. Until 2007, 28 areas had improved water in Tongyu, accounting for 93.33%. These 28 projects were well running, covering a population of 7980. One hundred and fouty-eight high arsenic areas had changed water, reaching a rate of 94.27%. One hundred and fouty-eight projects changedg water were running normally, benefiting a population of 46 214. In the surveyed spots, arsenic content was between 0.004 mg/L and 0.005 mg/L in 2006 and between 0.010 mg/L and 0.021 mg/L in 2007, all in the normal range to the time being. The arsenic contents in urine of the children aged 8-12 in the monitoring spots were determinted, averaging at 0.024 mg/L in 15 samples, fluctuating between 0.005 mg/L and 0.048 mg/L in 2006. The average content in urine from adults was 0.019 mg/L in 53 samples, fluctuating between 0.005 mg/L and 0.087 mg/L in 2007. The arsenic contents in urine were all in the normal range in 2006 and 2007. In 2006 14 endemic arsenism patients, all in light symptoms, had been checked out, the morbidity being 6.19%. In 2007, 17 patients who were all in light symptoms were identified in a rate of 6.94%. There was no significant difference of morbidity between the two years(χ2=0.1059, P>0.05). Two patients died, unrelating with drinking high arsenic water in 2006 and 2007. Conclusions The prevention and control measures are well implemented in Tongyu. The water-improving projects are functioning well. The condition of endemic arsenism is slight and hasn't changed so much in these two years. The arsenic contents in urine of children and adults within the normal range, showing that improving water can control the occurrence and the development of endemic arsenism.

Humanin(HN,its analogue [Gly14]-Humanin,HNG)was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults.But the relative low content of this peptide in its natural sources limits its further characterization.An expression vector pET32a/HNG was corstructed and transformed it into E.coli BL21 trxB(DE3).HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography.Subsequently,the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC.A 23 mg recombinant HNG(rHNG)from 1 L bacterial culture was purified.The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide.The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence.Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

Objective To explore role of 64-slices spiral CT in differetiation of acute chest pains.Methods Thirty six patients with acute chest pains were performed 64-slices spiral CT chest angiography.Two-dimensional and three-dimensional reconstruction was performed in all patients by means of multiplanar reconstruction(MPR)(coronal,sgittal oblique),curved planar reformation(CPR), maximum intensity projection(MIP),and volume rendering(VR).All images were blindly reading by two experienced radiologist.DSA were performed at the same time in 16 cases.Results The coronary artery branches,pulmonary artery and aortic artery in all patients were showed clearly,The acute myocardial infarction were showed in 10 cases,The pulmonary artery embolism in 14 cases,The aortic dissection in 6 cases respectively,The Coronary embolism in One case ,pneumothorax In One case The constrictive pericarditis in 1 case respectively.Normal findings in 4 cases.Conclusion 64-slices spiral CT is a useful and noninvasive examination in acute chest pain.

OBJECTIVETo observe the effect platelet antigen modification by mPEG-SPA with different molecular masses.

METHODSPlatelet CD42a was modified by 5 kD and 20 kD mPEG-SPA, respectively, and the fluorescence intensity of CD42a was detect by flow cytometry and the three-dimensional structure of CD42a simulated to analyze the distribution of lysine in CD42a molecule.

RESULTSAfter platelet CD42a modification by 5 kD and 20 kD mPEG-SPA, the fluorescence intensity of CD42a decreased sharply by 85.54% and 88.65%, respectively, and multiple lysine regions were identified on the surface of CD42a molecule.

CONCLUSIONBoth 5 kD and 20 kD mPEG-SPA allow useful modification of platelet CD42a, but 20 kD mPEG-SPA is more advantageous than 5 kD mPEG-SPA.

Blood Platelets ; chemistry ; Humans ; Molecular Weight ; Platelet Glycoprotein GPIb-IX Complex ; chemistry ; Polyethylene Glycols ; chemistry ; Succinimides ; chemistry

OBJECTIVETo observe the effect of copper on bone fracture healing.

METHODS160 growing chickens aged 70 days were fed in 32 cages, each consisting of five. The fracture with 1 approximately 2 mm defects at the bilateral radius of wings was created by osteotomy. The chickens were divided into four groups, 40 each. Every morning, chickens of the first group, served as controls, were fed orally 0.8% Na-CMC solution (sodium carboxymethyl cellulose), those of the other three groups were fed with copper-Na-CMC suspension (copper powder suspended in 0.8% Na-CMC) with different doses of 20, 40 and 80 mg Cu/kg body weight. Each group was sacrificed 14, 21, 28 and 35 days postoperatively. Liver was taken for analysis of Cu and Zn. Both radius were removed by dissection. CT was performed quantitatively for the ment of the gray values of the callus. The biomechanical properties of the healing radius were analyzed by a three point bending test. Afterwards, the contents of calcium, magnesium, phosphorus, copper, zinc, iron and hydroxyproline in the callus were determined.

RESULTSThe gray values of the callus increased along with the increase of copper dose and the duration of observation. 21 and 35 days after operation, the gray values of the callus were significantly higher in the high-Cu group than in the controls respectively (909 +/- 220 vs. 597 +/- 155; 973 +/- 100 vs. 763 +/- 179 HU, P < 0.05). The level of calcium and magnesium in the callus 35 days after operation was much higher in the mid and high Cu groups than in the controls respectively (Ca: 177 +/- 26.7, 176 +/- 20.5 and 137 +/- 34.7 mg/g; Mg: 2.98 +/- 0.57, 3.06 +/- 0.46 and 2.43 +/- 0.53 mg/g P < 0.05). The contents of hydroxyproline in the callus 35 days after operation were significantly higher in the high-Cu group than in the controls (34.23 +/- 1.96 vs. 32.17 +/- 1.93 mg/g respectively). The biomechanical properties of the repaired radius had a better improvement tendency in the three Cu-fed groups than in the controls.

CONCLUSIONCopper is helpful in fracture healing; however, the effect of copper on fracture healing needs further study.

Animals ; Bony Callus ; Chickens ; Copper ; Fracture Healing ; Zinc

OBJECTIVEDetermination of estrone (E1) levels has a significant meaning in evaluating physiological effect and diagnosing some diseases. In order to detect free E1 in biological fluids, a monoclonal antibody specific for E1 was prepared after the complete antigen of E1 was synthesized. The purified monoclonal antibody was fully characterized for later immunoassay.

METHODS3-O-carboxymethyl ether derivative of E1 was synthesized and in turn coupled to bovine serum albumin (BSA) to form complete antigen E1-BSA. A monoclonal antibody (McAb) specific for E1 was produced both in vitro and in vivo by a hybridoma anti-E1. Anti-E1 was prepared by fusion of SP2/0 murine myeloma cells with spleen cells isolated from immunized BALB/c mouse. The McAb was characterized by enzyme-linked immunosorbent assay (ELISA), SDS-PAGE and Western-blotting. The specificity of the immunoassay was investigated by determining the cross-reactions of E1 analogs when free E1 was detected by competitive indirect enzyme-linked immunosorbent assay (CI-ELISA).

RESULTSAnalysis revealed that anti-E1 McAb (E1-McAb) was of the IgG1 type, the molecular weight of E1-McAb was 164,000 daltons. The affinity constant of E1-McAb with coated complete antigen was 8.2 x 10(8) L/mol. The linear range for free E1 determined by CI-ELISA was 10 pg/mL-10 ng/mL. The detection limit was 21.4 pg/mL (defined as twice the standard deviation of the blank).

CONCLUSIONThe CI-ELISA developed with E1-McAb was both sensitive and specific. The prepared E1-McAb can be used in some immunoassays.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Affinity ; Antibody Specificity ; Blotting, Western ; Cross Reactions ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Estrone ; immunology ; Female ; Hybridomas ; immunology ; Mice ; Mice, Inbred BALB C