Objective To detect the expressions of nm23-H1 and heat shock protein 27 (HSP27) and their clinical significance on development and metastasis in non-small cell lung carcinoma (NSCLC).Methods 75 tumor tissues from patients with NSCLC were included as experimental group and 28 pulmonary benign lesion tissues were as control group.The expressions of nm23-H1 and HSP27 in patients with different clinical and pathological characters were detected by immunohistochemistry.Results nm23-H1 and HSP27 were mainly expressed in cytoplasm,the positive rates of nm23-H1 and HSP27 were significantly higher in the experimental group than that in control group [41.3 ％ (31/75) vs 7.1％ (2/28),x2 =10.946,P =0.001,80.0 ％ (60/75) vs 46.4 ％ (13/28),x2 =11.131,P =0.001].Compared with control group,the positive rate of HSP27 was correlated with the degree of tumor differentiation (x2 =4.191,P =0.041).nm23-H1 was related with HSP27 in lung cancer (r =0.284,P =0.013).Conclusion nm23-H1 and HSP27 are related to the occurrence and development of NSCLC.The joint detection of nm23-H1 and HSP27 should be helpful to the diagnosis and judge the biological behavior of NSCLC.

To silence the expression of K-RASAsn12 in human pancreatic cancer cell line by vector-based RNAi(RNA interference) technique,two single-strand DNA sequences encoding mutant-specific shRNA (short haipin RNA) for K-RASAsn12 were synthesized and then inserted into pSilenCircle. The recombinant plasmid was called pSC-K-RASAsn12. According to the same method, pSC-GFP encoding shRNA for GFP was gained. Both recombinant plasmids were transfected into human pacreatic cancer cell line AsPC-1 and BxPC-3. The expression level of K-RASAsn12 was detected by semi-quantitative RT-PCR and Western blot. The result indicated that the recombinant plasmid edcoding mutant-specific shRNA for K-RASAsn12 can inhibit significantly the expression of K-RASAsn12 without affection of wild-type K-RAS(K-RASWT)in Human Pancreatic Cancer Cell Line.

OBJECTIVE:To survey the dosage range,clinical effect and safety of hypertonic sodium chloride hydroxylethyl starch 40 injection(HSS40) on anti-hemorrhagic shock METHODS:The changes of blood pressure and laboratory examination indices were detected after HSS40 was infused into the patients who suffered from hemorrhagic shock RESULTS:HSS40 could rapidly raise the blood pressure in a dose below 500ml and the effective rate could reach 100% In part of patients,the levels of serum Cl- and Na+ increased temporarily and restored after 24 hours CONCLUSION:HSS40 was safe and effective in doses of 80ml～500ml The clinical doses of 300ml～500ml are recommended

OBJECTIVETo investigate the CT and MRI features of peripheral primitive neuroectodermal tumors (pPNETs) and evaluate its diagnostic value.

METHODSThe clinicopathological data of 9 surgically treated patients with peripheral primitive neuroectodermal tumors confirmed by pathology were collected, spiral CT (4/9) and MRI (6/9) plain scanning and dynamic enhancement scanning were performed preoperatively. Both CT and MRI scannings were performed in 1 case. Those CT and MR images were retrospectively reviewed and analyzed together with clinicopathological findings.

RESULTSThe 9 lesions were located in skeletal muscles (n = 6), pelvic cavity (n = 2) and thoracic cavity (n = 1). The tumor size was 7.4-18.3 cm in diameter with a mean diameter of 11.6 cm. The shape of those lesions was round or ellipse (4 lesions) and irregular (5 lesions). The tumor usually presented as ill-defined masses, with homogeneous (n = 2) or inhomogeneous density (n = 7). Seven cases, including the 3 lesions located in the chest and pelvis, showed obvious necrosis and multilocular cyst formation. The tumors showed iso-density as that of the adjacent muscles on CT plain scans and moderate heterogeneous enhancement after intravenous injection of contrast agents. The features of the tumors on the MRI including slightly low signal intensity on SE T1-weighted imaging, iso-signal intensity or slightly high signal intensity on FSE T2-weighted imaging and heterogeneous dynamic delayed contrast-enhancement with obvious necrosis in most of them. Six cases had a lesion in the skeletal muscles, presented as a giant ill-defined masse surrounding bone and extended along neural route with bone destruction to varying degrees.

CONCLUSIONPrimitive neuroectodermal tumor is a kind of malignant tumor with proliferation of small, undifferentiated neuroectodermal cells, usually occurring in children or adolescent and frequently located in the extremities, chest cavity, pelvic cavity and chest wall. It typically presents as a large, ill-defined masse extending along neural route with heterogeneous and obvious enhancement after intravenous injection of contrast agents. The tumors located in the chest and pelvic cavities and some in the extremities show obvious necrosis and multilocular cyst formation, while some of the tumors in the extremities appear as homogeneous solid masses.

Adolescent ; Adult ; Child ; Diagnostic Errors ; Female ; Histiocytoma, Malignant Fibrous ; diagnosis ; Humans ; Male ; Muscle Neoplasms ; diagnosis ; diagnostic imaging ; pathology ; Muscle, Skeletal ; Neuroectodermal Tumors, Primitive, Peripheral ; diagnosis ; diagnostic imaging ; pathology ; Pelvic Neoplasms ; diagnosis ; diagnostic imaging ; pathology ; Radiography ; Retrospective Studies ; Thoracic Neoplasms ; diagnosis ; diagnostic imaging ; pathology ; Young Adult

OBJECTIVETo investigate the correlation between genotypes and biochemical phenotypes of phenylalanine hydroxylase (PAH) in patients with phenylketonuria (PKU).

METHODSThirteen exons and flanking introns of PAH gene in 102 patients with high blood phenylalanine levels (Phe > 120 umol/L) at initial diagnosis were amplified with polymerase chain reaction and analyzed with single strand conformation polymorphism (SSCP), denaturing high performance liquid chromatography (DHPLC) and DNA sequencing. Correlation between genotypes and biochemical phenotypes was analyzed.

RESULTSBiochemical assaying has indicated that 69 patients had classical PKU (Phe> 1200 umol/L), 31 were moderate (Phe 600-1200 umol/L), and 2 were mild (Phe 400-600 umol/L). More than 41 mutations and 75 genotypes have been identified. There were 9 (8.8%) homozygous mutations, which included 3 cases with R111X/R111X, 1 case with IVS4-1G>A/IVS4-1G>A, 3 cases with R243Q/R243Q and 2 cases with V399V/V399V. Among these 8 belonged to classic PKU phenotypes, except for a R243Q/R243Q genotype which has led to a moderate phenotype. In 91 patients carrying compound PAH mutations, 61 were classic, 29 were moderate, and 1 was mild. Patients who were heterozygous for R111X/R243Q and EX6-96A>G(Y204C)/R243Q were found with both classic and moderate PKU phenotypes. Certain individuals who have carried 2 null mutant alleles such as R111X/V399V, EX6-96A>G/Y356X and EX6-96A>G/V399V only showed a moderate phenotype. Individuals with R111X/A165D and R176X/A165D genotypes, on the other hand, respectively presented moderate and classic PKU phenotypes.

CONCLUSIONNinety percent of our patients are compound heterozygotes. Independent assortment of mutant alleles has resulted in a complex genotype-phenotype correlation. Although in most cases a correlation may be found, caution should still be taken upon genetic counseling. The phenomena where similar or even identical genotype may give rise to different biochemical phenotypes have implied that other factors may also influence the phenylalanine metabolism.

Adolescent ; Alleles ; Child ; Child, Preschool ; Exons ; Female ; Gene Frequency ; Genetic Association Studies ; Genotype ; Humans ; Infant ; Infant, Newborn ; Introns ; Male ; Mutation ; Phenotype ; Phenylalanine Hydroxylase ; genetics ; metabolism ; Phenylketonurias ; genetics ; metabolism

OBJECTIVETo investigate the association between the polymorphism of serotonin transporter gene-linked polymorphic region(5-HTTLPR) and slow transit constipation(STC).

METHODSPolymerase chain reaction was used to assess 5-HTTLPR polymorphism of SERT gene in 54 patients with STC and 100 healthy controls.

RESULTSThe frequencies of serotonin transporter short/short(S/S) and allele S genotypes were significantly higher in STC patients than those in controls(72.2% vs 50.0%; 83.3% vs 72.5%; both P< 0.05). There were no significant differences in 5-HTTLPR polymorphism respectively between the two groups according to gender and age(less than 45 and more than 45 years old). The frequency of S/S genotype was higher in the patients with less than 40% of the ingested markers evacuated within 72 h than those with more than 40% evacuated(71.7% vs 42.6%, P< 0.05).

CONCLUSIONThe presence of 5-HTTLPR allele S may contribute to the pathogenesis of STC.

Adolescent ; Adult ; Aged ; Alleles ; Chronic Disease ; Constipation ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Serotonin Plasma Membrane Transport Proteins ; genetics ; Young Adult

This study was aimed to investigate a novel MLAA-22 antigen derived from a U937 cDNA library by the SEREX approach and search for gene expression in various samples. Bioinformatic analysis was performed to forecast MLAA-22 information mined from databases and experimental datasets. CTL epitope was predictied and the specific antibody for MLAA-22 was elicited by using peptide-microspheres and adjuvants. Furthermore, SYBR Green real-time PCR and immunoblotting method were used to evaluate the specificity of gene expression. The results showed that the full length cDNA of MLAA-22 located on chromosome 17q11.2 was 2.0 kb in size, and a putative protein was approximately 72.4 kD for 631 amino acids. The MLAA-22 encoded a cancer/testis antigen in human, which is nonsecreting type, plasmosin, labile protein, hydrophilia, thermostability, and without signal peptide. Many motifs might related to growth, proliferation, differentiation, and apoptosis. Antigenic peptides was synthesized as the antigen with Fmoc/PyBOP method. Rabbits were immunized by injecting the synthetic peptide-KLH to obtain antibody and the immune sera analyzed with ELISA were 1:8000. SYBR Green real-time PCR and Western blot showed that MLAA-22 presented with a higher number of copy messages in M(5), lower in CML, but not in gastric carcinoma, renal carcinoma, LNCaP cell lines and normal adult tissues, etc. It is concluded that mlaa-22 is a novel acute monocytic leukemia-associated antigen gene and be extended to further discovery.
Amino Acid Sequence ; Base Sequence ; Cell Line, Tumor ; Chromosomes, Human, Pair 17 ; genetics ; Computational Biology ; Epitopes ; immunology ; Humans ; Leukemia, Monocytic, Acute ; genetics ; immunology ; Molecular Sequence Data ; Neoplasm Proteins ; genetics ; immunology

Objective To know the effect of nanometer titanium dioxide (nano -TiO2 )on the oxidative stress in zebrafish. Methods A total of 64 zebrafishes were randomly divided into 4 groups with different concentrations of nano -TiO2 exposure,which including control group,low dose group (2 mg/L),moderate dose group (8 mg/L)and high dose group (32 mg/L).The content of MDA,the activities of SOD and ATPase in gill,liver and muscle were measured in 1 0 zebrafishes in each group after 30 d exposure.Histopathological changes of gill,liver and muscle in the remainder fishes were observed.Results The content of Malondialdehyde (MDA)in the fish gill of the moderate and high dose group were significantly higher than that of the control group (P <0.05).The activity of Superoxide dismutase (SOD)in the fish gill of the high dose group was significantly increased compared with the control group (P <0.05).The activities of Adenosine triphosphatase(ATPase)in the fish gill of the moderate and high dose group were significantly decreased compared with the control group (P <0.05).The changes on the content of MDA and the activities of SOD in the fish liver were similar with that in the fish gill after nano -TiO2 exposure,while there was no significant change in the activity of ATPase of the fish liver.And no significant changes were observed in the content of MDA,the activities of SOD and ATPase in the fish muscle (P >0.05 ). Histopathological observation showed infiltration by inflammatory cells, swelling and degeneration, vacuolation and necrosis in zebrafish gill and liver with different degrees.Conclusion Nano -TiO2 exposure could induce the oxidative stress in zebrafish,causing pathological changes in the fish gill and liver.

Objective To explore the mutagenic effect of sodium pentachlorophenate (NaPCP) on zebrafish p53 gene coding sequence(CDS) in somatic cell.Methods The experiment was carried out using tuebingen strain of zebrafish, according to the results of acute toxicity test to determine the exposure levels in zebrafish.Zebrafish were randomly divided into blank control group and exposed groups, each containing 10 zebrafish.After exposing for 45d of NaPCP, the RNA was extracted from liver of zebra fish, and the p53 gene including a complete coding sequence of was obtained by RT-PCR.Results LC50 of NaPCP was 18.4 μg/L.Sequence analysis showed that the p53 gene CDS length of 1125bp, encoding 374 amino acids.The percent identity between the published zebrafish sequence of p53 (GI:425876786)and ours was 99.2%,with the other biological sequence of p53 existing some differences.After 45d exposure, zebrafish p53 gene of NaPCP exposure group had mutated at the concentration of 1.8 μg /L.The base substitution of GAG→AAG at codon 8,CAT→CAG at codon 148 and CAG→CAA at codon 229 were detected by PCR-directed sequencing.This may result in the Glu→Lys and His→Gln of expressed p53 protein.Conclusion NaPCP is a kind of gene mutation, which can induce the mutation of p53 gene in zebrafish somatic cells, that has the potential mutagenic risk for humans.

Objective To evaluate the safety of the Eucommia folium and to provide the basis of the rational use ofEucommia folium resource.Methods According to < Technical Standards for Testing & Assessment of Health Food>,version 2003, acute safety test/micronucleus test of born marrow in mice/sperm shape abnormality test in mice/Ames test andthirty-day feeding test were all performed after water extract concentrations of Eucommia folium treatment (1 mLconcentrations equal to 1.5 g Eucommia folium) . Results On the acute toxicity test, the MTD of rats and mice were both40.0 mL/kgbw. The results of genetic toxicity test were all negative in the dosage of 10 g/kg bw(according to the dosage ofEucommia folium) .The results of thirty-day feeding test indicated that there have no significant differences in body weight,blood cytology indexes, blood biochemical indexes, organ/body ratio and pathematology examination in the rats in all thegroups (0.83, 1.67, 3.30 mL/kg) as compared to the control group(P>0.05) . Conclusion Under this experimentalcondition, no obvious abnormity and toxic reaction were observed.