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.

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 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.

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.

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

The clinical manifestations of five children with paroxysmal kinesigenic dyskinesia (PKD) were retrospectively analyzed and their gene mutations were analyzed by high-throughput sequencing and chromosome microarray. The 5 patients consisted of 4 males and 1 female and the age of onset was 6-9 years. Dyskinesia was induced by sudden turn movement, scare, mental stress, or other factors. These patients were conscious and had abnormal posture of unilateral or bilateral extremities, athetosis, facial muscle twitching, and abnormal body posture. The frequency of onset ranged from 3-5 times a month to 2-7 times a day, with a duration of <30 seconds every time. Electroencephalography showed no abnormality in these patients. Three patients had a family history of similar disease. The high-throughput sequencing results showed that a heterozygous mutation in the PRRT2 gene, c.649_650insC (p.R217PfsX8), was found in two patients; the mutation c.436C>T (p.P146S) was found in one patient; a splice site mutation, IVS2-1G>A, was found in one patient. The two mutations c.436C>T and IVS2-1G>A had not been reported previously. The chromosome microarray analysis was performed in one patient with negative results of gene detection, and the chromosome 16p11.2 deletion (0.55 Mb) was observed. Low-dose carbamazepine was effective for treatment of the 5 patients. PKD is a rare neurological disease. The detection of the PRRT2 gene by multiple genetic analysis can help the early diagnosis of PKD.
Carbamazepine ; therapeutic use ; Child ; Chromosome Deletion ; Chromosomes, Human, Pair 16 ; Dystonia ; complications ; diagnosis ; drug therapy ; genetics ; Electroencephalography ; Female ; Humans ; Male ; Membrane Proteins ; genetics ; Mutation ; Nerve Tissue Proteins ; genetics

BACKGROUNDThe solitary pulmonary nodule (SPN) is one of the most common findings on chest radiographs. The objectives of clinical practice are to differentiate malignant nodules from benign nodules in the least invasive way and to make a specific diagnosis. This study was aimed to evaluate the correlation between perfusion imaging features and microvessel density (MVD) and vascular endothelial growth factors (VEGF) in SPNs using multi-slice computed tomography (MSCT); and to provide the theoretical basis for SPN blood flow pattern and blood flow quantitative features. Also, the study called for the discussion of the method's clinical application value in the differential diagnosis of benign and malignant SPNs.

METHODSSixty-eight patients with SPN underwent multi-location dynamic contrast enhanced (nonionic contrast material was administrated via the antecubital vein at a rate of 4 ml/s) MSCT. Precontrast and postcontrast attenuations on every scan was studied. Perfusion, peak height, and the ratio of the peak height of the SPN to that of the aorta were analyzed. Perfusion was calculated using the maximum gradient of the time-density curves (TDC) and the peak height of the aorta. The quantitative parameters (perfusion, peak height, ratio of peak height of the SPN to that of the aorta) of the blood flow pattern were compared with MVD and the VEGF expression of immunohistochemistry.

RESULTSThe perfusion peak heights of malignant ((96.15 +/- 11.55) HU) and inflammatory ((101.15 +/- 8.41) HU) SPNs were significantly higher than those of benign ((47.24 +/- 9.15) HU) SPNs (P < 0.05, P < 0.05). Ratios of SPN-to-aorta of malignant and inflammatory SPNs were significantly higher than those of benign SPNs (P < 0.05, P < 0.05). No significant differences were found between the peak height and SPN-to-aorta ratio of malignant SPNs and inflammatory SPNs (P > 0.05, P > 0.05). The precontrast densities of inflammatory SPNs were lower than those of malignant SPNs (P < 0.05). Perfusion values of malignant and inflammatory SPNs were significantly higher than those of the benign SPNs (P < 0.05, P < 0.05). The VEGF positive expressions appeared in 32 patients with malignant SPNs and 2 patients with benign SPNs, and the average value of the MVD was higher in patients with malignant SPNs (36.88 +/- 6.76) than in patients with either benign (4.51 +/- 0.60) or inflammatory (26.11 +/- 5.43) SPNs (P < 0.05, P < 0.05). There were statistically significant correlations between the CT perfusion feature and the MVD. The highest correlation was between the peak height of SPN and the MVD (r = 0.657, P < 0.05).

CONCLUSIONSTumor microvessel density and VEGF expression facilitate the exploration of the pathophysiological basis of CT perfusion in SPNs. Multi-slice CT perfusion has shown strong positive correlations with angiogenesis in SPNs.

Adult ; Aged ; Female ; Humans ; Immunohistochemistry ; In Vitro Techniques ; Lung ; diagnostic imaging ; metabolism ; pathology ; Male ; Microvessels ; pathology ; Middle Aged ; Neovascularization, Pathologic ; Perfusion Imaging ; Solitary Pulmonary Nodule ; diagnostic imaging ; metabolism ; pathology ; Tomography, X-Ray Computed ; methods ; Vascular Endothelial Growth Factor A ; analysis