Objective To improve the recognition of nervous system symptoms of inborn errors.Methods Five patients with organic acidemias were screened by urine organic acid analysis(gas chromotography-mass spectrometry,GC/MS),3 cases of methylmalolic acidemias(MMA) and 2 cases of propionic acidemias(PA) were confirmed.They were treated with special diet and medicine after diagnosis.Result The improvement of mental development was observed after treatment.Conclusions Most of organic acidemias involve nervous systems,causing non-specific symptoms of nervous system as lethergy,seizures,mental retardation.Inborn errors of metabolism shall be kept in mind when causes of the symtoms of acidosis,seisures,mental retardation and lethergy are investigated.GC/MS is a very important method in diagnosis of organic acidemias.Early diagnosis and early treatment can improve the mental prognosis.

Objective:To analyze the activities of human NKG2D ligand MICA/B promoter induced by 5-Fu.Methods:The 5'-end flanking regions of MICA /B promoter and their different truncated fragments were amplified from A549 genome by PCR. The resulting amplicons were cloned into pGL3-Basic vector to generate the MICA/B luciferase reporter plasmids. All the constructs were transiently transfected A549 cells. The promoter region activities were determined by dual-luciferase reporter assays. The effect of 5-Fu on the promoter activities of MICA/B was also tested.Results and Conclusion:The 5'-end flanking regions of MICA /B promoter and five of their different truncated fragments were successfully obtained. The normalized luciferase reporter gene activities driven by the above promoters and fragments were 3.61,2.26,1.63,0.313,0.711 and 0.663 for MICA and 17.49,10.11,7.398,0.822,0.997 and 0.49 for MICB,respectively. Promoter activities in transiently transfected A549 cells treated by 20,40,80,160 and 320 μg/m of 5-Fu increased 1.69,1.48,1.62,1.55 and 1.78 fold for MICA and 1.44,1.87,1.38,1.19 and 1.25 fold for MICB. Our results suggest that 5-FU can significantly up-regulate the promoter activity of both MICA and MICB.

Increasing attention has been focused on the antitumor activity of artemisinin derivatives in recent years, for artemisinin had been reported to have cytotoxic effects against HL-60, P388 and MCF-7 tumor cells. We report here the synthesis and evaluation for antitumor activity of a series of artemisinin-ether derivatives bearing tetrahydropyrrole, morpholine, piperidine, substituted piperidines and azoles with various linkers. Sixteen 10-O-substituted dihydroartemisinin derivatives were designed and synthesized, all of which have never been reported in literatures and whose antiproliferative effects on human breast cancer MCF-7, MCF-7/Adr and HL-60 cells were determined by MTT assay or direct cell counting. Each of these artemisinin derivatives possessed better effects than dihydroartemisinin evidently against HL-60 and MCF-7 cells growth, while less potent than doxorubicin. All target compounds exhibited significantly improved potency compared to DHA and doxorubicin on the doxorubicin-resistant MCF-7/Adr cells, so did they in their sensitive counterparts MCF-7 cells. Among them, compounds GF02, GH04 and ZH04 showed strong activity against these three cell lines growth. Further research is undergoing.
Antineoplastic Agents ; chemical synthesis ; chemistry ; Artemisinins ; chemical synthesis ; chemistry ; Breast Neoplasms ; pathology ; Cell Proliferation ; Doxorubicin ; Drug Design ; HL-60 Cells ; drug effects ; Humans ; MCF-7 Cells ; drug effects

OBJECTIVE: To study the clinical efficacy of callotasis for pathological segmental defects in child femur. METHODS: Thirty-nine patients with segmental femoral defects suffered from suppurative osteomyelitis were treated with the bilateral, unilateral external fixation frame or intramedullary callotasis. The rolongational rate was from 2 to 2.5 cm per month after the operation. RESULTS: After a 13 to 57 month follow-up, all cases were healed according to clinical examination and radiological observation. The prolongational length ranged from 9 to 31 centimeters. Average ratio of prolongation reached 49%. The healing index was 30 d/cm, and the healing time of non-union was 134 days. No recurrent suppurative osteomyelitis or fracture was observed. CONCLUSION Callotasis is a reliable, simple and hyperadaptable method for femural segmental defects.
Adolescent ; Bone Lengthening ; methods ; Bony Callus ; surgery ; Child ; Child, Preschool ; Female ; Femur ; surgery ; Follow-Up Studies ; Humans ; Male ; Osteomyelitis ; surgery ; Suppuration

OBJECTIVETo observe the effects of fosinopril (FOS) on proliferation and secretion of extracellular matrix of rat glomerular mesangial cell induced by LPS.

METHODSIn vitro culture method for glomerular mesangial cells (GMC) of rat was established and passages 3 - 10 of the cells were used in the experiment after identification. The experiment included the following 5 groups: control group (Ctrl), LPS group (LPS), high, medium and low dose FOS groups (FOS1, FOS2 and FOS3 groups, respectively). GMC proliferation was detected by methyl thiazolyl tetrazolium (MTT) incorporation method at 24 and 48 h; the changes of laminin (LN), fibronectin (FN) and ColIV protein secretion was detected by the enzyme-linked immunosorbent assay (ELISA). The changes of LNbeta(2) mRNA expression was detected by semi-quantitative real-time RT-PCR.

RESULTS(1) LPS could induce the mesangial cell proliferation, FOS inhibited this effect of proliferation induced by LPS. (2) Mesangial cells could secrete some extracellular matrix (ECM) protein in normal culture medium, mesangial cell secreted ECM protein was significantly higher in LPS group than that in Ctrl group (P < 0.01), but significantly lower in all FOS groups than that in LPS group (P < 0.01). (3) Mesangial cell could express LNbeta(2) mRNA in normal culture medium, LNbeta(2) mRNA expression was significantly higher in LPS group than that in Ctrl group at all time points, but was significantly lower in FOS group than that in LPS group.

CONCLUSIONSLPS could induce increased secretion of the ECM, including LN, FN, ColIV; FOS could inhibit the secretion of ECM in GMC in a dose-dependent manner at mRNA and protein levels.

Animals ; Cell Proliferation ; Cells, Cultured ; Extracellular Matrix Proteins ; secretion ; Fosinopril ; pharmacology ; Gene Expression Regulation ; Lipopolysaccharides ; Mesangial Cells ; drug effects ; metabolism ; Rats

OBJECTIVETo study the distribution of mutations of UDP-glucuronosyltransferase 1A1 (UGT1A1) gene and its relationship with hyperbilirubinemia among neonates with hyperbilirubinemia of Guangxi Heiyi Zhuang nationality.

METHODSTotal genomic DNA was extracted from the blood of 100 neonates with hyperbilirubinemia (case group) and 100 neonates without hyperbilirubinemia (control group), all of whom were selected from Guangxi Heiyi Zhuang population. TATA box and all exons of UGT1A1 gene were amplified by PCR and directly sequenced.

RESULTS(TA)7 insertion mutation in TATA box, G71R missense mutation in exon 1, and 4 single nucleotide polymorphisms (SNPs) (rs199539868, rs114982090, rs1042640 and rs8330) in exon 5 were observed. The allele frequency of G71R mutation in the case group was significantly higher than that in the control group (P<0.01). There were no significant differences in the genotype distribution and allele frequency of TATA box mutation and SNPs (rs1042640 and rs8330) between the two groups (P>0.05). The logistic regression analysis showed that the odds ratios (95% confidence intervals) of UGT1A1 TATA box mutation, G71R mutation, and SNPs (rs1042640 and rs8330) associated with the development of neonatal hyperbilirubinemia were 0.846 (0.440, 1.629), 3.932 (1.745, 8.858), 0.899 (0.364, 2.222), respectively.

CONCLUSIONS(TA)7 insertion mutation and G71R missense mutation of UGT1A1 gene are common mutation types in neonates with hyperbilirubinemia of Guangxi Heiyi Zhuang nationality. Four SNPs (rs199539868, rs114982090, rs1042640, and rs8330) was first reported in China. UGT1A1 G71R missense mutation is a risk factor for hyperbilirubinemia in neonates of Guangxi Heiyi Zhuang nationality.

China ; ethnology ; Glucuronosyltransferase ; genetics ; Humans ; Hyperbilirubinemia, Neonatal ; genetics ; Infant, Newborn ; Logistic Models ; Mutation ; Polymorphism, Single Nucleotide ; TATA Box

Objective To test the possibility of reciprocal pathways between medullary visceral zone (MVZ) and hypothalamic paraventricular nucleus (PVN) or supraoptic nucleus (SON) following hyperosmotic stimulation. Methods Hyperosmotic pressure animal model was established by administering3% sodium chloride as drinking water to rats. The distributions and expressions ofHRP retrogradely labeled neurons, Fos, tyrosine hydroxylase (TH) or vasopressin (VP) positive neuron and lial fibrillary acidic protein (GFAP) positive astrocytes (AST) in MVZ, SON and PVN were observed by quadruple labeling methods of WGA-HRP retrograde tracing combined with anti-Fos, TH (or VP) and GFAP immunohistochemical technique. Results Fos positive neurons within the MVZ, PVN and SON increased markedly. There were also a large number of GFAP positive structures in the brain and their distribution pattern was fundamentally similar or analogous to Fos positive neurons in the above-mentioned areas. The augmented GFAP reactivities took on hypertrophic cell bodies, thicker and longer processes.Quadruplicate immunohistochemical staining showed that a neuron could be closely surrounded by many AST and they formed neuron-astrocytic complex (N-ASC). Conclusion The neurons and AST might be very active following hyperosmotic pressure and N-ASC as a functional unit might serve to modulate the osmotic pressure. There was reciprocal osmoregulation pathways between the MVZ and SON or PVN in the brain.

OBJECTIVE: To analyze the polymorphisms of glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) gene exon 14, GPI-PLD activity of leucocyte in the peripheral blood,and the relationship in leukemia patients of Han nationality in Hunan. METHODS: Both 96 leukemia patients and 96 healthy persons of Han nationality in Hunan were researched [including 48 acute non-lymphocytic leukaemia (ANLL) patients as group A, 31 acute lymphoblastic leukaemia (ALL) patients as group B, 12 chronic granulocytic leukaemia (CML) patients as group C, 5 chronic lymphocytic leukaemia (CLL) patients as group D]. The polymorphisms were analyzed by PCR-SSCP and sequencing;. and GPI-PLD activities were determined by GPI-anchored placental alkaline phosphatase (PLAP) as substrate and triton-X114 partioning. RESULTS: There were four variations in the coverage of GPI-PLD gene exon 14 of leukemia patients and healthy persons. The codons of variation were: 1257 C-->T, 1298 T-->C, 1218 C-->A, 1257 C-->A. The total various frequency in leukemia patient and healthy person, which was determined by SSCP, was 28.12% and 20.83%. On the basis of the percentage of GPI-anchored PLAP conversion, the leucocyte GPI-PLD activities of the 96 leukemia patients were measured. Compared with the 96 healthy controls, the leukocyte GPI-PLD activites of ANLL and CLL patients were significantly increased; the acticities of ALL and CML patients were significantly reduced. CONCLUSION Leukocyte GPI-PLD gene in the peripheral blood, which belongs to healthy persons and leukemia patients of Han nationality in Hunan, is polymorphism. The leukocyte GPI-PLD activities in the four groups are remarkable.
Adolescent ; Adult ; Aged ; Base Sequence ; Exons ; genetics ; Female ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; Phospholipase D ; genetics ; metabolism ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Single-Stranded Conformational ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics

OBJECTIVE: To construct the recombinant adeno-associated virus(rAAV) vector plasmid pSNAV2.0-TK containing HSV1-TK gene, to produce recombinant adeno-associated virus rAAV2/HSV1-TK, and to detect the integration and expression of HSV1-TK gene in lens epithelial cells transfected by rAAV2/HSV1-TK, and to provide foundation for gene therapy of posterior capsular opacification. METHODS: The recombinant vector plasmid constructed by gene recombinant technology was analyzed by PCR and restriction enzyme digestion. The cell strain BHK-21/TK was screened by G418 after the plasmid was transfected into BHK-21 cells,with the helper virus HSV1-rc/UL2 to produce the recombinant virus rAAV2/HSV1-TK. The purity of rAAV2/HSV1-TK was detected by SDS-PAGE and HPLC, and the titre of rAAV2/HSV1-TK was observed by dot blot hybridization. The HSV1-TK gene in lens epithelial cells transfected by rAAV2/HSV-TK was investigated by PCR and RT-PCR. RESULTS: The recombinant plasmid proved successful by PCR and restriction enzyme digestion. The recombinant virus rAAV2/HSV1-TK was produced successfully and its titre was 1 x 10(12) v.g./mL by dot blot hybridization. The HSV1-TK gene was integrated and expressed in lens epithelial cells. CONCLUSION The recombinant adeno-associated virus vector plasmid containing HSV1-TK gene is successfully constructed, and high titre recombinant adeno-associated virus (rAAV2/HSV1-TK) is obtained. The HSV1-TK gene in lens epithelial cells is expressed after being transfected by rAAV2/HSV1-TK.
Animals ; Cloning, Molecular ; Cricetinae ; Dependovirus ; genetics ; metabolism ; Epithelium, Corneal ; cytology ; metabolism ; Genetic Vectors ; Herpesvirus 1, Human ; enzymology ; genetics ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Thymidine Kinase ; biosynthesis ; genetics ; Transfection

Objective To establish a method for detection of the human papi 11 omavirus(HPV)6b E7-specific antibodies in serum and cervical secretion from patients with condyloma acuminatum(CA).Methods A full-length HPV 6b E7 gene was amplified by PCR from the CA tissue to construct the recombinant plasmid pET32a(+)/HPV 6b E7.The expression of prokaryotic protein was analyzed by SDS-PAGE and Western blot,then purified with Ni-NTA Agarose affinity column and used as an diagnostic antigen for establishing indirect ELISA method,to detect specific serum IgG and specific cervical secretion slgA from 56 CA patients,81 healthy control.Sera from 43 cervical cancer was served as control.HPV 6b DNA from 56 CA patients was identified by PCR.Results Data showed that the nucleotide homology of cloned sequence was 99.5%,compared to the standard sequences of HPV 6b E7 gene(GenBank accession number:NC001355).A high level expression of E7 fusion protein was obtained in prokaryotic expression system(40 μg/ml).Based on HPV 6b E7 fusion protein being used as coating antigen,results from ELISA showed that the absorbance rates(A)of serum IgG from CA,cervix cancer and healthy control groups were 1.82±0.48,1.36 ± 0.39 and 1.39 ± 0.27,respectively.The level of IgG antibody in the serum of CA group was significantly higher than that in cervix cancer group and healthy control(P＜0.05).The A values of cervical secretion sIgA in CA and healthy control groups were 0.63 ± 0.26 and 0.53 ± 0.06,respectively,while the level of sIgA antibody in the cervical secretion of CA group was also significantly higher than that in healthy controls(P＜0.05).The positive rate of HPV 6b E7 DNA in CA tissue was 78.6%(44/56)by PCR method.When compared the results measured by PCR,the HPV 6b E7-specific IgG and sIgA antibodies by ELISA used to detect the patients infected with HPV 6b infection,showed that the sensitivity rates were 68.2%(30/44)and 54.6%(24/44)respectively,and the specificity were all 100%(12/12).Conclusion Based on the serum and cervical secretion specific HPV 6b E7 antibodies from patients with CA to diagnose HPV 6b infection,results showed medium sensitivity and high specificity,and could further be used to investigate the epidemiological characteristics of HPV 6b infection.