OBJECTIVETo explore the molecular basis for a CD36 deficiency individual and distribution of CD36 gene mutation in Guangxi population.

METHODSA female individual was studied. CD36 phenotype was detected by monoclonal antibody immobilization of platelet antigens assay (MAIPA) and flow cytometry (FCM). The coding regions of the CD36 gene were sequenced. A DNA-based polymerase chain reaction-sequence specific primer (PCR-SSP) assay was used to verify the identified mutation. Cell lines expressing the mutant and wild-type CD36[CD36(MT) and CD36(WT)] were established, with the expression of CD36 determined by Western blotting. The distribution of CD36 gene mutation was investigated among 1010 unrelated individuals with the PCR-SSP assay.

RESULTSBoth MAIPA and FCM assays showed that the patient had type II CD36 deficiency. DNA sequencing showed that she has carried a heterozygous mutation T538C (Trp180Arg) in the exon 6 of CD36. Sequencing of cDNA clone confirmed that there was a nucleotide substitution at position 538 (538T>C). Western blotting also confirmed that the CD36 did not express on the CD36(MT) cell line that expressed the 538C mutant, but did express on the CD36(WT) cell line. The novel CD36 mutation T538C was further verified with 100% concordance of genotyping results by DNA-based PCR-SSP assay and 1010 unrelated individuals. No CD36 538C allele was detected among the 1010 individuals.

CONCLUSIONThis study has identified a novel CD36 mutation T538C(Trp180Arg)(GenBank: HM217022.1), and established a genotyping method for the novel sequence-specific primer PCR. The novel mutation is rare in Guangxi and can cause type II CD36 deficiency.

Base Sequence ; Blood Platelet Disorders ; genetics ; Blood Platelets ; cytology ; metabolism ; Blotting, Western ; CD36 Antigens ; genetics ; metabolism ; Cells, Cultured ; DNA Mutational Analysis ; DNA Primers ; genetics ; Exons ; genetics ; Female ; Flow Cytometry ; Fluorescent Antibody Technique ; Genetic Diseases, Inborn ; genetics ; Genotype ; Genotyping Techniques ; methods ; Humans ; Middle Aged ; Monocytes ; cytology ; metabolism ; Mutation, Missense ; Polymerase Chain Reaction ; methods

Objective To evaluate the correlation between multi-slice spiral CT(MSCT)perfusion imaging parameters and gastric carcinoma differentiation.Methods MSCT perfusion parameters including blood flow(BF),patlak blood volume(PBV),time to peak (TTP)and patlak permeability surface(PPS)were obtained in 44 patients with pathologically confirmed gastric carcinoma before sur-gery.Patients were divided into two groups;26 well differentiated and 18 poorly differentiated.According to the extent of cell differ-entiation ,BF,PBV,TTP and PPS were investigated in two groups.Results Parameters in the well differentiated gastric carcinoma group BF 101.8±6.7 (mL·min-1 ·100 mL-1 ),PBV 85.9±3.4 (mL/1 000 mL ),TTP 121.6±1 1.5(0.1 s),PPS 82.7±1 1.0 (0.5 mL·min-1 ·100 mL-1 )respectivily,Parameters in the poorly differentiated gastric carcinoma group,BF 105.3 ±7.7,PBV 92.1±7.1,TTP 113.0±10.5,PPS 94.3±22.9 respectivily.There was no statistic difference in BF value between the two groups(P >0.05). There were statistic differences in TTP、PBV and PPS between the two groups(P <0.05).Conclusion MSCT perfusion parameters may reflect the degree of differentiation of gastric carcinoma or a certain extent in vivo.

BACKGROUNDCytochrome b5 reductase 2 (CYB5R2) is a potential tumor suppressor that inhibits cell proliferation and motility in nasopharyngeal carcinoma (NPC). Inactivation of CYB5R2 is associated with lymph node metastasis in NPC. This study aimed to explore the mechanisms contributing to the anti-neoplastic effects of CYB5R2.

METHODSPolymerase chain reaction (PCR) assays were used to analyze the transcription of 84 genes known to be involved in representative cancer pathways in the NPC cell line HONE1. NPC cell lines CNE2 and HONE1 were transiently transfected with CYB5R2, and data was validated by real-time PCR. A chick chorioallantoic membrane (CAM) embryo model was implanted with CYB5R2-expressing CNE2 and HONE1 cells to evaluate the effect of CYB5R2 on angiogenesis. An immunohistochemical assay of the CAM model was used to analyze the protein expression of vascular endothelial growth factor (VEGF).

RESULTSIn CYB5R2-transfected NPC cells, PCR assays revealed up-regulated mRNA levels of Fas cell surface death receptor (FAS), FBJ murine osteosarcoma viral oncogene homolog (FOS), phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1), integrin beta 3 (ITGB3), metastasis suppressor 1 (MTSS1), interferon beta 1 (IFNB1), and cyclin-dependent kinase inhibitor 2A (CDKN2A) and down-regulated levels of integrin beta 5 (ITGB5), insulin-like growth factor 1 (IGF1), TEK tyrosine kinase (TEK), transforming growth factor beta receptor 1 (TGFBR1), and VEGF. The angiogenesis in the CAM model implanted with CYB5R2-transfected NPC cells was inhibited. Down-regulation of VEGF by CYB5R2 in NPC cells was confirmed by immunohistochemical staining in the CAM model.

CONCLUSIONCYB5R2 up-regulates the expression of genes that negatively modulate angiogenesis in NPC cells and down-regulates the expression of VEGF to reduce angiogenesis, thereby suppressing tumor formation.

Animals ; Carcinoma ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Chickens ; Cytochrome-B(5) Reductase ; Down-Regulation ; Gene Regulatory Networks ; Genes, Tumor Suppressor ; Humans ; Nasopharyngeal Neoplasms ; Neovascularization, Pathologic ; Oxidoreductases ; Real-Time Polymerase Chain Reaction ; Transfection ; Up-Regulation ; Vascular Endothelial Growth Factor A

Objective To evaluate the clinical value of MRA on the abnormal vessels in Moyamoya and cerebral revascularization , and to evaluate PWI in the observation of cerebral hemodynamics before and after cerebral revascularization.Methods Twenty-four patients with Moyamoya disease ascertained by DSA received cerebral revascularization on one side.MRA and PWI were performed for all patients before and after the operation , while DSA was performed after operation in nine patients to compare the images of MRA and DSA by three experienced radiologists.Perfusion parameters in terminal branches of middle cerebral artery ( MCA) on the operative side were compared with those on the contralateral sides and the cerebellum , including cerebral blood flow (CBF), cerebral blood volume (CBV), mean transit time (MTT), and delay time ( DT) , as well as relative ratio ( values on the operative side/values on the contralateral side , and values on the operative side/values on the cerebellar region ) of perfusion parameters (rCBF, rCBV, rMTT and rDT) were calculated.Comparisons of the data between different groups were performed using paired Student′s t test.Results MRA was similar to DSA in displaying the internal carotid artery , main branches stenosis, Moyamoya vessels, and cerebral revascularization.DSA was better in displaying collateral vessels than MRA, but primary images of MRA provided anatomic and pathologic information of cerebral parenchyma.After the operations , rCBF (1.30 ±0.27) and rCBV (1.26 ±0.21) of MCA perfusion regions on the operative and the contralateral sides were higher than rCBF (0.73 ±0.15) and rCBV (0.98 ±0.12) before the operation significantly (t=-7.19,-6.64,P<0.05).rMTT (1.06 ±0.20) and rDT (1.07 ± 0.18) after the operation were lower than rMTT(1.53 ±0.34)and rDT (1.40 ±0.26) before the operation (t=5.62,5.40,P<0.05) .In MCA perfusion regions on the operative and cerebellar sides , rCBF(1.93 ± 0.34)and rCBV(2.25 ±0.35)were higher than rCBF(0.88 ±0.18)and rCBV(1.16 ±0.22)(t=-3.04,-3.06,P<0.05) before the operation.rMTT (1.13 ±0.29) and rDT (1.29 ±0.12) were lower than rMTT (1.88 ±0.19 ) and rDT ( 3.29 ±0.47 ) before the operation ( t =4.01, 4.72, P <0.05 ).Conclusions MRA is a safe , reliable method to evaluate abnormal vessels and cerebral revascularization after the operation in moyamoya disease.PWI can detect poor cerebral perfusion , and can be used for assessment of the effect of cerebral revascularization.

[Objective] To explore the best method for sequelae neuralgia of herpes zoster.[Method] Randomly divide the said patients into groups treated with external application of Chinese drugs and sky moxibustion,make observation and comparison between them.[Result] Both got high late curative effect,however most patients in the sky moxibustion group were cured without recurrence,the pain relieving rate was high once being cured,the reduce of pain index was apparent after treatment,which was of statistical meaning compared with drug application group.[Conclusion] The sky moxibustion method combines drugs and moxibustion well and has marked advantages in treating herpes zoster,worth of promotion and application.

The aim of this study is to evaluate the effect of Interleukin-6 on cyclophosphamide-induced hematopoietic damnification. The doses of Interleukin-6 in 3 different regimens were hypodermally injected into dogs for 7 days respectively to establish the cyclophosphamide-induced hematopoietic damnification model. The effect of Interleukin-6 on the production of platelets and the amount of other cells in the dogs' bone marrow were determined on the 21st day. The results showed that Interleukin-6 significantly alleviated the reduction of platelet count and recovered the platelets level faster. The impedance effects of Interleukin-6 directed against hematopoietic damnification of bone marrow and spleen were shown by pathological examination. These suggest that the Interleukin-6 can significantly impede cyclophosphamide-induced hematopoietic damnification.
Animals ; Bone Marrow Cells ; drug effects ; metabolism ; Cyclophosphamide ; adverse effects ; Dogs ; Female ; Interleukin-6 ; pharmacology ; therapeutic use ; Leukopenia ; chemically induced ; prevention & control ; Male ; Thrombocytopenia ; chemically induced ; prevention & control

Objective: FF456 is a new gene cloned in our lab, which belongs to c-type G protein-coupled receptors and has high homology to the olfactory receptor family. Northern blotting and RT-PCR showed that the expression of FF456 was exclusively restricted in liver and was significantly down-regulated in hepatoma. In an effort to study the function of FF456 gene, high titer antibody is indispensable. Methods:Full-length cDNA of FF456 was reconstructed into pcDNA3.1(-) vector, which was used for immunizing 6- to 8- weeks old female BALB/c mice. The effects of different injection manners, solvents and dosages on the antibody production have been compared. For detecting the titer and specificity of the antisera produced by DNA immunization, a peptide containing FF456 specific sequence was expressed by E.coli expression system. Results:Finally specific antisera with titers as high as 1 ∶ 50 000 was obtained. Conclusion:Immunofluorescence assays showed FF456 was expressed in hepatocytes with high tissue specificity. The FF456 expression in hepatoma cells was decreased dramatically.