Objective To clone and express the HIV-1B gp120 genes isolated at different organizations from a patient died of AIDS dementia complex (ADC) in eukaryotic cells.Methods Using the genomic DNA isolated from peripheral lymphnodes,choroid plexus and occipital white matter from a patient died of ADC as the template,HIV-1B gp120 gene was amplified with PCR.After sequenced,HIVIB gp120 was inserted into pcDNA3.1 ( + ) and recombinant expressing vector gp120/pcDNA3.1 ( + ) was constructed succeffuly confirming with sequencing. Then expressing vector was transfected into eukaryotic cells U87 using liposome transfection and expression of HIV-1B gp120 gene was assayed with indirect immunofluorescence.Results HIV-I B gp120 genes isolated from peripheral lymphnodes,choroid plexus and occipital white matter of the ADC patient were successfully cloned and recombinant expressing vector gp120/pcDNA3.i ( + ) could express envelope glycoprotein HIV-1B gp120 in U87 cells.Conclusion All the HIV-1B gp120 gene isolated at the different organizations of the same ADC patient could express in U87 cells,which may supply a valuable basis for studying the neurotoxicity and neurotoxic mechanism of HIV-1 gp120 protein.

OBJECTIVETo study the chemical constituents of Potentilla chinesis and their anticancer activities.

METHODChemical constituents were isolated by repeated column chromatography (Toyopearl HW-40C and preparative HPLC). The structures were elucidated on the basis of spectral data analysis. The isolated compounds were screened with two anticancer models.

RESULTFifteen triterpenes, alpha-amyrin (1) , beta-amyrin (2) , ursolic acid (3) , corosolic acid (4), euscaphic acid (5) , pomolic acid (6) , tormentic acid (7), 2alpha, 3alpha-dihydroxyurs-12-en-28-oic acid (8), 2beta, 3beta, 19alpha-trihydroxyurs-12-en-28-oic acid (9), asiatic acid (10) , 24-hydroxy tormentic acid (11) , myrianthic acid (12), oleanolic acid (13), maslinic acid (14) and 2alpha, 3alpha-dihydroxyolean-12-en-28-oic acid (15) , were isolated from P. chinesis.

CONCLUSIONCompounds 1, 2, 4 -15 were isolated from the plant for the first time. Compounds 4, 8 - 10, 12, 14 and 15 show anticancer activities. Compounds 4, 9 show strong activities.

Animals ; Antineoplastic Agents, Phytogenic ; chemistry ; isolation & purification ; pharmacology ; Cell Line ; Cell Survival ; drug effects ; Chromatography, High Pressure Liquid ; Fibroblasts ; cytology ; drug effects ; HeLa Cells ; Humans ; Mice ; Molecular Structure ; Plants, Medicinal ; chemistry ; Potentilla ; chemistry ; Triterpenes ; chemistry ; isolation & purification ; pharmacology

Objective To determine the clinical features,neurophysiological characteristics and cervical magnetic resonance imaging of sensory neumnopathy,and to describe the pathology of skin nerve, sural nerve and spinal dorsal columns.Methods Two patients who died from sensory neuron disease (SND)after infection of digestive tract were discussed including clinical features and ancillary tests which included neurophysiology and pathology of peripheral nerve and spinal dorsal columns.Associated documents are reviewed.Results Early ataxia,widespread sensory symptoms and global loss of deep tendon reflex were the distinctive signs of SND,which was characterized by non-length-dependent abnormalities of sensory nerve action potentials,a hallmark of ganglionopathies.The second patient showed normal cervical magnetic resonance imaging possibly because of short course of disease,while diffuse hyperintensity in the spinal posterior columns of SND was reported.Demyelination of spinal posterior columns and loss of mostly large diameter nerve fibers without regeneration clusters were the main pathological features.Conclusions The distinctive clinical features and neurophysiological characteristics of SND indicate that peripheral sensory nerve fibers are widely damaged.Pathology of spinal posterior columns confirm that central sensory pathway are impaired which allow the localization of the pathologic site to the dorsal root ganglion neurons.Cervical spinal MRI of SND are possibly normal at early phase.

OBJECTIVETo explore the mechanism of scalp catgut embedding for treatment of Parkinson's disease.

METHODSParkinson's disease model rats were prepared, and randomly divided into a model group, scalp acupuncture group, medication group, with a control group set up. The scalp acupuncture group were treated with catgut embedding at "the chorea-trembling conroued area" lateral to "Baihui" (GV 20) and the medication group with intra-gastric perfusion of L-dopa suspension. After treatment for 30 days, behavior indexes were investigated and malondialdehyde (MDA) and nitric oxide (NO) contents and superoxide dismutase (SOD) activity in the brain were detected.

RESULTSMDA content in the brain of the model rats were significantly decreased (P < 0.01), NO level significantly increased (P < 0.05) by scalp catgut embedding, with no significant change of SOD activity (P > 0.05); and the behavior indexes and tissue and form of the brain had significant improvement.

CONCLUSIONScalp catgut embedding has regulative action on anti-oxidant enzyme system in the Parkinson's disease rats, has significant improvement of cells, tissues and form of the brain.

Acupuncture Therapy ; methods ; Animals ; Catgut ; Female ; Male ; Malondialdehyde ; analysis ; Nitric Oxide ; analysis ; Parkinson Disease ; metabolism ; therapy ; Rats ; Rats, Wistar ; Scalp ; Superoxide Dismutase ; metabolism

OBJECTIVETo compare therapeutic effects of red-hot needle therapy and filiform needle therapy on nodules of breast of the type of stagnation of liver-qi and phlegm coagulation.

METHODSSix hundred cases were randomly divided into a treatment group and a control group, 300 cases in each group. The treatment group were treated with red-hot needle pricking at the proliferative parts and Ashi points as main, and the control group with filiform needle therapy.

RESULTSIn the treatment group, 240 cases were cured, 58 cases improved and 2 cases were not cured with an effective rate of 99.3%; and in the control group, 113 cases were cured, 165 cases improved and 22 were not cured with an effective rate of 92.7%, with a significant difference between the two groups (P = 0.001).

CONCLUSIONThe therapeutic effect of red-hot needle therapy is better than that of filiform needle therapy on nodules of breast of the type of stagnation of liver-qi and phlegm coagulation.

Acupuncture Therapy ; methods ; Adult ; Female ; Fibrocystic Breast Disease ; etiology ; therapy ; Humans ; Middle Aged

Objective To study the clinical significance of traditional Chinese medical psychological nursing in the therapy of chlossma patients.Methods Totals of 103 patients were randomly divided into control group (n =48) that received the conventional nursing,and treatment group (n =55) that received the traditional Chinese medical psychological nursing on the bose of routine nursing.Then,the treatment compliance and treatment time was observed and compared between two groups.Results The treatment compliance of treatment groups was significantly better than that of control group (37/55 vs 22/48 ; Z =2.07,P ＜ 0.05).The treatment group had significantly longer treatment time than control group [(2.65 ± 0.43)months vs (1.13 ±0.36)months; t =19.29,P ＜0.01].Conclusions The traditional Chinese medical psychological nursing can improve the compliance of chloasma patients,and prolong their treatment time.So,it has important clinical significance for the therapy of chloasma patients.

BACKGROUNDHIV-1 infected and immune-activated macrophages and microglia secrete neurotoxins, such as tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), which play major role in the neuronal death. It has been shown that different HIV-1 variants have varying abilities to elicit secretion of TNF-α by peripheral blood mononuclear cell (PBMC); however, whether the difference of gp120 gene could affect the secretion of TNF-α and IL-1β by glial cells is unknown. The aim of this study was to explore the association between gene diversity and induction of neurotoxic cytokines.

METHODSIn this study, we constructed retroviral vectors MSCV-IRES-GFP/gp120 using HIV-1 gp120 genes isolated from four different tissues of one patient who died of AIDS dementia complex (ADC). Recombinant retroviruses produced by cotransfection of MSCV-IRES-GFP/gp120, pCMV-VSV-G and pUMVC into 293T cells were collected and added into U87 glial cells. Concentrations of TNF-α and IL-1β secreted by transduced U87 cells were assayed with ELISA separately.

RESULTSThe four HIV-1 gp120 were in the different branch of the neighbor-joining tree. Compared to the pMIG retrovirus (gp120-negative) or U87 cells, all the gp120-positive recombinant retroviruses induced more TNF-α (P < 0.01) and IL-1β (P < 0.01). In addition, compared with the L/MIG retrovirus, all the three brain gp120-positive recombinant retroviruses induced less TNF-α (P < 0.01) and IL-1β (P < 0.01).

CONCLUSIONSHIV-1 gp120 could induce U87 cells secret more TNF-α and IL-1β again. The more important is that difference of HIV-1 gp120, especially cell-tropism may account for the different ability in eliciting secretion of TNF-α and IL-1β, which might supply a novel idea helping understand the pathogenesis of ADC.

AIDS Dementia Complex ; metabolism ; virology ; Cell Line ; Cell Line, Tumor ; Enzyme-Linked Immunosorbent Assay ; HIV Envelope Protein gp120 ; genetics ; metabolism ; Humans ; Interleukin-1beta ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective To study the diversity of HIV-1 tat gene in CNS and peripheral tissue of a patient with ADC and a patient with non-ADC, so as to research HIV evolution, the mechanism of CNS invasion and the pathogenesis of ADC. Methods The tat gene was amplified with nested PCR from genomic DNA which was extracted from spleen and basal ganglia of one non-ADC patient with a wide range of cerebral artery atherosclerosis and one ADC patient. PCR products were cloned into the PGEM-T vector, after transformation and selection by ampicillin and blue/white spotting. Five of positive clones were sequenced.HIV-1 tat sequences were processed with BioEdit and MEGA4. With the softwares, neighbor-joining tree, pdistances, values of ds/dn, and analysis of amino acid motifs were all done, so as to research the diversity of HIV-1 tat gene in CNS and peripheral tissue. Results Gene mutation of HIV-1 tat exist in the two patients,the mutation process of tat isolated from ADC patient suffered more compartmentalization than tat isolated from non-ADC patient, the differences of tat genes between CNS and peripheral tissue in ADC patient were greater than the non-ADC patient. Ds/dn showed that the virus gene mutation played a major role, the body intend to remove harmful non-synonymous mutations. ConclusionsThe compartmentation of tat gene in CNS and peripheral tissue of the two patients was different, the reason may be related to the pathway of HIV into the CNS, the relationship between HIV gene mutation in CNS and ADC still need more investigation.

Aged ; Female ; Hepatitis C, Chronic ; complications ; Humans ; Interferons ; administration & dosage ; adverse effects ; therapeutic use ; Liver Cirrhosis ; drug therapy ; etiology ; Male ; Middle Aged ; Treatment Outcome

A laccase gene (lacD) from the basidiomycete Trametes sp. 420 was heterologously expressed in Pichia pastoris in two ways, resulting in two recombinant enzymes of rLacDx with native N-terminus and rLacDe with eight additional amino acid residues at N-terminus. The yields of rLacDx and rLacDe in shaken-flask cultures after an 18-day growth were 1.21 x 10(5) u/L and 7.38 x 10(4) u/L, respectively, as determined with 2,2'-azinobis(3-ethylbenzothia-zoline- 6-sulfonic acid) (ABTS) as substrate. The yield of rLacDx was further increased to 2.39 x 10(5) u/L under high-density fermentation while the production process was decreased to 7.5 days. In addition, rLacDx and rLacDe exhibited similar enzymatic characters in oxidizing substrate guaiacol, and were stable at 50 degrees C and at a pH range from 3 to 10. However, the specific activity of rLacDx (1761 u/mg) for ABTS was higher than that of rLacDe (1122 u/mg), and the apparent Km value of rLacDx (427 microM) was less than that of rLacDe (604 microM).
Cloning, Molecular ; Fermentation ; Isoenzymes ; biosynthesis ; genetics ; Laccase ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Trametes ; enzymology ; genetics