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Biomedical subjects

R Lu

Publications and source records attributed to R Lu.

At least 37 records · Page 2Linked to original sources

[Single particle source apportionment of atmospheric aerosol based on proton microprobe].

A proton microprobe with high resolution and high sensitivity was applied to analyze atmospheric aerosol in the people's park in Shanghai urban for single particle source apportionment. The results showed that the sum of the average contribution rates from the sources of soil dust, building construction dust, vehicle exhaust and metallurgical industry, which are the main pollution sources of atmospheric aerosol in Shanghai urban and surpasses 80%. Besides, about 13% of the particles could not be identified. On the basis of cluster analysis of these particles, they are preliminary known belonging to two different individual sub-pollution sources and a new unidentified pollution source.

Aerosols↗

[Studies on cloning and expression of hepatitis B virus x gene].

OBJECTIVE: To express and purify the x gene of hepatitis B virus (HBV), and then the purity of the protein could specifically be analysed by the human antibodies against HBV x antigen (protein). METHODS: HBV x gene was amplified from the complete genome by PCR, and cloned in the pGEX-2T fused expression vector. After transforming the plasmid into E.coli, the recombinant with HBx gene was obtained. The fusion protein was generated with E.coli fused expression system and purified with affinity chromatography. RESULTS: The HBx protein could be applied to detecting specific antibody of hepatitis B virus patients. CONCLUSIONS: The purity of the HBx protein can specifically be recognized by the human antibodies against HBV x antigen.

Base Sequence↗

[Different serum lipid adjustment drugs for the treatment of hyperlipidemic fatty liver].

OBJECTIVE: To research the effect of different serum lipid adjustment drugs, Simvastatin and Fenofibrate, on hyperlipidemic fatty liver. METHODS: The rat models of hyperlipidemic fatty liver induced by high fat diet were treated with Fenofibrate and Simvastatin respectively to investigate the change of the liver index, liver function, the serum and liver lipid, MDA, and pathologic histology. RESULTS: The liver index and the content of triglyceride (TC1.80 +/-0.20/2.10+/-0.33), total cholesterol (TG 0.76+/-0.18/1.09+/-0.31), MDA both in the serum and the liver in the model group were significantly higher than those in control group. Slight or moderate hepatic steatosis was observed in model group. Fenofibrate decreased serum triglyceride, total cholesterol, and MDA both in serum and liver significantly, but increased the ALT, ALP, liver index, liver lipids, and finally leading to severe hepatic steatosis. Simvastatin lowered the level of lipid and MDA both in serum and liver, and liver histology was improved nearly to normal. Moreover, little side effect on the liver was observed. CONCLUSIONS: Simvastatin can be applied for prevention and treatment of hyperlipidemic fatty liver safely and effectively, while Fenofibrate can not.

Animals↗

[Analysis of the nucleotide sequence for C and NS5 regions and the genotype of HCV isolate in Shandong Province].

OBJECTIVE: To study the genotype of HCV epidemic strains in Shandong Province. METHODS: HCV fragments of C(432 bp) and NS5(319 bp) regions were amplified by RT nested PCR. The fragments were cloned to T vector and sequenced by dideoxynucleotide chain termination reaction. RESULTS: Three of all 4 strains in C region were 1b genotype, and 1 was 2a. All of 10 strains of NS5 region were 1b genotype. No other genotype was found in this region. The homology of nucleotide sequence was over 90% compared with that of more than 50 strains in Gen Bank. CONCLUSIONS: The major genotype of HCV epidemic strains in Shandong Province is 1b. The 2a genotype is also found in this area.

Genotype↗

[Physico-chemical properties and biological activities of a glycoconjugate SPPA-1 from Spirulina platensis].

AIM: To isolate polysaccharide from Spirulina platensis and determine its sugar position, molecular weight and biological activities. METHODS: Sephadex G-75 and CM-Sephadex C-50 were used. The sugar position was analyzed by gas chromatography, the molecular weight was determined by GPC. The homogeneity of this glycoconjugate was determined by HPLC and CE. IR and NMR spectra were used to determine the glycosidic linkage. RESULTS: SPPA-1 was a homogeneous glycoconjugate, its carbohydrate chain was composed of alpha-glucopyranan, carbohydrate content is 91.70%, Nitrogen content was 0.96%, the molecular weight was 69.00 x 10(4), SPPA-1 could eliminate O2-. radicals. CONCLUSION: SPPA-1 was an antioxidative glycoconjugate from Spirulina platensis.

Amino Acids↗

Adeno-associated viral vector-mediated vascular endothelial growth factor gene transfer induces neovascular formation in ischemic heart.

Vascular endothelial growth factor (VEGF) plays important roles in physiological and pathological angiogenesis. Recent studies have demonstrated that direct injection of VEGF protein, plasmid DNA, or an adenoviral vector encoding the VEGF gene into ischemic myocardium or limb can induce collateral blood vessel formation and improve perfusion of the ischemic areas. However, these approaches have limitations ranging from a short-lasting effect to angioma formation. In this study, we investigated the feasibility of using adeno-associated viral (AAV) vectors to deliver VEGF genes to mouse myocardium. A cytomegalovirus promoter was used to drive genes for a human VEGF isoform, VEGF(165), and LacZ. A mouse myocardial ischemic model was generated by ligation of the anterior descending coronary artery. Approximately 10(11) copies of the AAV-VEGF vector mixed with 10(10) copies of AAV-LacZ were injected to one site of normal myocardium and a total of 10(11) copies of AAV-VEGF were injected to multiple sites of myocardium around the ischemic region. LacZ gene expression was observed up to 3 months after the vector inoculation. After AAV-VEGF inoculation, neoangiogenesis was observed in the ischemic heart model but not in normal heart tissue. An inflammatory-cell infiltration was not observed in the AAV-VEGF- and AAV-LacZ-inoculated hearts, and angioma-like structure was not observed. These results indicated that injection of the AAV vector directly to myocardium could mediate efficient gene transfer and transgene expression and that VEGF gene delivered by AAV vector can induce angiogenesis in ischemic myocardium.

Animals↗

Regulation of the promoter activity of interferon regulatory factor-7 gene. Activation by interferon snd silencing by hypermethylation.

The molecular mechanism by which virus induces expression of the early inflammatory genes has not yet been completely elucidated. Previous studies indicated that the virus-mediated transcription of type I interferon (IFN) genes required activation of two members of IFN regulatory factor (IRF) family, IRF-3 and IRF-7, where the expression of IRF-7 was found to be indispensable for the induction of IFNA genes. To determine the factors that regulate expression of IRF-7 gene, as well as its inducibility by type I IFNs, we have isolated and characterized the promoter and first intron of the human IRF-7 gene. This region shows a presence of two potential interferon-sensitive response elements (ISRE/IRF-E). However, only the ISRE present in the first intron was functional and conferred interferon inducibility in a transient transfection assay. Using a pull-down assay with an oligodeoxynucleotide corresponding to this ISRE immobilized to magnetic beads, we have demonstrated that this ISRE binds ISGF3 complex and IRF-1 from the extract of IFN-treated cells but not from the untreated cells. We have further shown that the previously observed lack of expression of IRF-7 in 2fTGH fibrosarcoma cell line, correlated with hypermethylation of the CpG island in the human IRF-7 promoter. The repression of the promoter activity was relieved by treatment with DNA methyltransferase inhibitor 5-aza-deoxycytidine. In vitro methylation of IRF-7 promoter silenced IRF-7 directed expression of luciferase gene in HeLa cells that express endogenous IRF-7 gene. Whether silencing of IRF-7 by methylation is instrumental for the process of tumorigenesis remains to be determined.

Azacitidine↗

Zhangfei: a second cellular protein interacts with herpes simplex virus accessory factor HCF in a manner similar to Luman and VP16.

Host cell factor (HCF, C1, VCAF or CFF) is a cellular protein that is required for transcription activation of herpes simplex virus (HSV) immediate-early (IE) genes by the virion protein VP16. The biological function of HCF remains unclear. Recently we identified a cellular transcription activator, Luman. As with VP16, the transactivation function of Luman is also regulated by HCF. Here we report a second human protein, Zhangfei (ZF) that interacts with HCF in a fashion similar to Luman and VP16. Although ZF shares no significant sequence homology with Luman, the two proteins have some structural similarities. These include: a basic domain-leucine zipper (bZIP) region, an acidic activation domain and a consensus HCF-binding motif. Unlike Luman, or most other bZIP proteins, ZF by itself did not appear to bind consensus bZIP-binding sites. It was also unable to activate promoters containing these response elements. Although in transient expression assays ectopically expressed ZF was unable to block transactivation by VP16 of a HSV IE promoter, ZF could prevent the expression of several HSV proteins in cells infected with the virus. The ability of ZF to block the synthesis of the HSV IE protein ICP0 relied on its binding to HCF, since a mutant of ZF that was unable to bind HCF was also unable to prevent viral IE protein expression.

5' Untranslated Regions↗

Inhibition of PC cell-derived growth factor (PCDGF, epithelin/granulin precursor) expression by antisense PCDGF cDNA transfection inhibits tumorigenicity of the human breast carcinoma cell line MDA-MB-468.

PC-cell derived growth factor (PCDGF) is an 88-kDa growth factor originally purified from the highly tumorigenic teratoma PC cell line and corresponds to the epithelin/granulin precursor. In teratoma cells, PCDGF expression was shown to be essential for tumorigenicity. We have reported that PCDGF was expressed in estrogen receptor-positive (ER(+)) human mammary epithelial cells in an estrogen-dependent fashion. In this study, we have investigated PCDGF expression in human mammary epithelial cell lines ranging from immortalized nontumorigenic cells to ER(+) and ER(-) breast carcinoma cells. Northern and Western blot analyses indicated that PCDGF mRNA and protein expression was low in nontumorigenic cells and increased in human breast carcinomas cell lines in a positive correlation with their tumorigenicity. Treatment of the ER(-) MDA-MB-468 cells with anti-PCDGF neutralizing antibody resulted in a dose-dependent inhibition of their proliferation, suggesting that secreted PCDGF acted as an autocrine growth factor for breast carcinoma cells. We then examined the in vitro and in vivo growth properties of MDA-MB-468 cells, where PCDGF expression had been inhibited by antisense PCDGF cDNA transfection. Inhibition of PCDGF expression resulted in a reduced proliferation rate in vitro and a 60-80% reduction in colony formation. Tumor formation in vivo was dramatically inhibited in antisense cells with a 90% inhibition of tumor incidence and tumor weight. These results demonstrate the importance of PCDGF overexpression for the proliferation and tumorigenicity of ER(-) breast carcinomas and suggest that PCDGF overexpression may play an important role in human breast cancer.

Breast Neoplasms↗

Adeno-associated viral-mediated gene transfer to hepatoma: thymidine kinase/interleukin 2 is more effective in tumor killing in non-ganciclovir (GCV)-treated than in GCV-treated animals.

Interleukin 2 (IL-2) enhancement of herpes simplex virus-thymidine kinase (HSV-TK)/ganciclovir (GCV)-induced tumor killing was studied by cloning the human interleukin 2 gene into an HSV-TK-bearing adeno-associated viral (AAV) vector (TK/IL-2). The mouse hepatocellular carcinoma cell line Hepa 1-6 was used as a model in this study. We found that TK/IL-2-transduced Hepa 1-6 cells were more susceptible to ganciclovir treatment than tumor cells transduced with only TK in both nude mice and immunocompetent C57L/J mice. TK/IL-2-transduced tumors also showed shrinkage without GCV treatment. The tumor-killing effect of AAV-mediated TK/IL-2 gene transfer was further studied by inoculating animals with TK/IL-2- or TK-transduced tumor cells mixed with unmodified cells with or without GCV treatment. Although tumor growth in each group was inhibited, the best result was obtained from the TK/IL-2-transduced group without GCV treatment. In this group, 10% of the transduced tumor cells could eradicate the whole tumor in 50% of the animals tested as well as provide long-term protection against tumor cell rechallenge. When this group was treated with GCV, the antitumor effect of TK/IL-2 was reduced. We attribute this to the early ablation of transgene-bearing tumor cells by GCV treatment, which thus reduces the duration of IL-2 expression. We conclude that (i) TK/IL-2 plus GCV treatment generates a stronger tumor-killing effect than HSV-TK plus GCV and (ii) tumor killing of TK/IL-2 is more effective in non-GCV-treated animals than in GCV-treated animals.

Animals↗

A functional cloning strategy, based on a binary PVX-expression vector, to isolate HR-inducing cDNAs of plant pathogens.

We have devised a novel, high-throughput functional cloning method to isolate cDNAs from plant pathogens of which the products elicit a hypersensitive response (HR) in plants. Copy DNA, made from RNA isolated from the tomato pathogen Cladosporium fulvum grown under nutrient-limiting conditions in vitro, was cloned into a binary, potato virus X (PVX)-based expression vector and transformed to Agrobacterium tumefaciens. 9600 colonies were individually toothpick-inoculated onto leaflets of tomato plants resistant to C. fulvum. Four cDNAs were identified whose expression induced formation of a necrotic lesion around the inoculation site. One of these clones, specifically inducing HR on tomato plants carrying the Cf-4 resistance gene, encodes race-specific elicitor AVR4. The other three cDNAs, inducing a non-genotype-specific HR, encode a protein highly homologous to bZIP, basic transcription factors. To determine whether this approach has general applicability, part of the library was also inoculated onto Nicotiana tabacum var. Samsun NN, which is not a host for C. fulvum. Four independent HR-inducing cDNAs were identified which all encode ECP2, an extracellular protein of C. fulvum known to induce necrosis in certain Nicotiana species. These observations confirm that this functional screening method is a versatile strategy to identify cDNAs of pathogens that encode (race-specific) elicitors and other HR-inducing proteins.

Agrobacterium tumefaciens↗

Affinity purification and partial characterization of the zonulin/zonula occludens toxin (Zot) receptor from human brain.

The intercellular tight junctions (TJs) of endothelial cells represent the limiting structure for the permeability of the blood-brain barrier (BBB). Although the BBB has been recognized as being the interface between the bloodstream and the brain, little is known about its regulation. Zonulin and its prokaryotic analogue, zonula occludens toxin (Zot) elaborated by Vibrio cholerae, both modulate intercellular TJs by binding to a specific surface receptor with subsequent activation of an intracellular signaling pathway involving phospholipase C and protein kinase C activation and actin polymerization. Affinity column purification revealed that human brain plasma membrane preparations contain two Zot binding proteins of approximately 55 and approximately 45 kDa. Structural and kinetic studies, including saturation and competitive assays, identified the 55-kDa protein as tubulin, whereas the 45-kDa protein represents the zonulin/Zot receptor. Biochemical characterization provided evidence that this receptor is a glycoprotein containing multiple sialic acid residues. Comparison of the N-terminal sequence of the zonulin/Zot receptor with other protein sequences by BLAST analysis revealed a striking similarity with MRP-8, a 14-kDa member of the S-100 family of calcium binding proteins. The discovery and characterization of this receptor from human brain may significantly contribute to our knowledge on the pathophysiological regulation of the BBB.

Amino Acid Sequence↗

Potential role for luman, the cellular homologue of herpes simplex virus VP16 (alpha gene trans-inducing factor), in herpesvirus latency.

The cascade of herpes simplex virus (HSV) gene expression that results in viral replication begins with the activation of viral immediate-early (IE) genes by the virion-associated protein VP16. VP16 on its own is inefficient at associating with complexes formed on IE gene promoters and depends upon the cellular factor HCF for its activity. In this respect VP16 mimics the host basic leucine zipper (bZIP) protein Luman, which also requires HCF for activating transcription. Our objective is to explore interactions between Luman and HCF and to determine if they play a role in the biology of herpesviruses. In this report we show that in cultured cells ectopically expressed Luman was retained in the cytoplasm, where it colocalized with Calnexin, a protein normally associated with the endoplasmic reticulum (ER). Retention of Luman in the ER depends on a hydrophobic segment of the protein that probably serves as a transmembrane domain. Deletion of this domain changed the intracellular location of Luman so that most of the mutant protein was in the nucleus of cells. While HCF was present in the nucleus of most cells, in cells expressing Luman it was retained in the cytoplasm where the two proteins colocalized. This cytoplasmic association of Luman and HCF could also be demonstrated in neurons in trigeminal ganglia removed from cattle soon after death. Cells in tissue culture that expressed Luman, but not a mutant form of the protein that fails to bind HCF, were resistant to a productive infection with HSV type 1 (HSV-1). We hypothesize that similar Luman-HCF interactions in sensory neurons in trigeminal ganglia result in the suppression of viral replication and the establishment of latency. Interestingly, Luman could activate the promoters of IE110 and LAT, two genes that are critical for reactivation of HSV-1 from latency. This suggests a role for Luman in the reactivation process as well.

Amino Acid Sequence↗

[Design and implementation of a landscape analysis system based on remote sensing data].

Because of the functions of Geographic Information System(GIS) being not satisfied to the requirements of landscape analysis, a landscape analysis system was designed with Visual C++ and demonstrated with remote sensing data. This system adopted the method f picking-up patch boundary pixel set, improved the patch searching speed, and supplied some landscape analysis functions, such as the calculation of the distance between patches, and the boundary length of neighboring patch, which are not provided by current GIS. It was used to analyzed the landscape of suburbs in northwest Beijing, and the shortcomings of current GIS applied in landscape analysis were overcome.

Algorithms↗

[CT guided percutaneous fine needle aspiration for early diagnosis of pancreatic infection after acute necrotizing pancreatitis].

OBJECTIVE: To assess CT-guided percutaneous fine needle aspiration (FNA) for early diagnosis of pancreatic infection after acute necrotizing pancreatitis. METHODS: CT-guided percutaneous FNA was employed to diagnose pancreatic infection after acute necrotizing pancreatitis in 14 patients. The aspirates were smeared, cultured and subjected polymerase to chain reaction (PCR) for detecting bacteria. RESULTS: Ten patients were diagnosed as having pancreatic infection and 4 non-pancreatic infection. FNA was totally performed for 15 times: left retroperitoneal route for 12 times, left and right retroperitoneal route for one time, and transperitoneal route for two times. The sensitivity of smear, PCR, culture to diagnose pancreatic infection was 7/10, 9/10, 10/10, respectively. Their specificity was 4/4, and the time for obtaining the results was 30 minutes, 4 hours, 3 days respectively. CONCLUSIONS: Retroperitoneal approach can be used for most patients with acute necrotizing pancreatitis when performing CT guided-percutaneous FNA. PCR of aspirates is a rapid and accurate method to diagnose pancreatic infection.

Adult↗

[A Chinese herbal compound induces neurotrophy in astrocyte by a new study method called "cerebrospinal fluid pharmacology in Chinese materia medica"].

OBJECTIVE: To find a effective method by which the potency of Chinese materia medica (CMM) on the central nervous system (CNS) could be correctly investigated. METHOD: The cerebrospinal fluid(CSF) and serum after oral CMM 1 to 1.5 hours was gained and then added into the astrocyte cell culture medium. After 48 h, the above medium was in proportion transferred to the neuron culture medium. Typan blue staining, LDH and MTT test were utilized to observe the drug effect on the neuron survival. RESULTS: The serum, but not the CSF, showed the toxicity on the astrocyte. MTT test on the neuron survival showed that the OD492nm value in the CMM group was higher than the other groups (P < 0.05). The astrocyte medium stimulated by the CSF including CMM could promote the neuron axon growth. CONCLUSION: If observing the CMM potency on CNS in vitro, the CSF was better than the serum, we called it "Cerebrospinal Fluid Pharmacology in Chinese Materia Medica". Furthermore, the CMM could induce the neurotrophy of astrocyte.

Animals↗

[Herbological study for the Beimu categorical Chinese medicine on the original plants].

This article reports the herbological study of "Beimu" categorical Chinese medicine on the original plants. The result showed that the medicinal "Beimu" before "Weijin" period is "Jia Beimu" (Bolbostemma paniculatum); "Nenbei" daynasty began to used "Zhe Beimu" (Fritillaria thunbergii), but "Jia Beimu" and "Hubei Beimu" (F. hupehensis) occupied an important position continuously; Last phase of "Ming" dynasty began to used the "Zhe Beimu" and "Chuan Beimu" (F. cirrhosa, F. unibracteato, F. przewalskii and F. delavayi) for "Beimu" greatly. The "Zhe Beimu" from Xiangshan, Zhejiang Province, is named "Xiang Beimu" and others are named "Tu Beimu" and the "Chuan Beimu" included the "Yi Beimu" (F. walujuewii and F. pallidiflora) continuously. So that, authors suggested that the contemporary documents such as Chinese Pharmcopoeia must recover the name "Beimu" to change the name "Tu Beimu" so as to avoid chaos with the "Tu Beimu" of the herbological documents; "Hubei Beimu" may take the place of "Zhe Beimu" but don't substitue for "Chuan Beimu"; "Yi Beimu" may be merged into "Chuan Beimu" to gear to actual circumstances and history; "Ping Beimu" to take the place of "Chuan Beimu" is not foundation from herbological documents.

Drug Contamination↗