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Biomedical subjects

Yan Zeng

Publications and source records attributed to Yan Zeng.

At least 37 records · Page 2Linked to original sources
PubMed2005

[Establishment of the three-dimensional finite element model of the first permanent mandibular molar and its stress analysis].

OBJECTIVE: To establish more rational three dimensional finite element model (FEM) of the first permanent mandibular molar and to study the internal stress distribution. METHODS: The FEM was applied to analyze the stress distribution of the first permanent mandibular molar under various loading conditions. RESULTS: Under all five loading conditions, the maximum von-Mises stress and compressive stress in enamel were larger than that in dentin, and the tensile stress in enamel was smaller than that in dentin. When vertical force and buccal-lingual oblique force were loaded, the stresses in the enamel and dentin were minimum; when lingual-buccal oblique force was loaded, the stresses were maximum; the stresses were mainly distributed at cervical region, furcation and apical area of the mesial-lingual root canal wall. CONCLUSIONS: The stress value changes and stress distribution in the enamel and dentin were closely related with the direction of the occlusion force.

Adult↗
PubMed2004

[Evaluation of neck moving ability after anterior cervical fusion].

OBJECTIVE: To observe the relationship of adjacent segment degeneration and neck symptom after anterior cervical fusion. METHODS: Making follow-up to 66 cases who had accepted anterior cervical fusion for 1 - 16 years, average 10.5 years. Observe the mobilization and gliding degree of adjacent segment in flexion-extension X-ray film, neck symptom, and the relationship of them. In 59 of these patients, the mobilization of entire cervical spine and non-fusion segment was observed before/after operation. RESULTS: The incidence of adjacent segment instability in follow-up term was 72.7%, and 40.9% of all patients had significant neck symptom. The rate of significant neck symptom in patients who had instability was 48%, whereas which in patients without instability was 18.8%, and the difference was statistically significant (P < 0.05). The difference of mild instability and severe instability was not statistically significant (P > 0.1). The mobilization of entire cervical spine decreased obviously (P < 0.001). The mobilization of adjacent segment increased obviously (P < 0.01), and which of non-adjacent segment had no change (P > 0.05). CONCLUSION: Majority of patients who have accepted anterior cervical fusion possessed instability of adjacent segment, but many of them have no symptoms. Adjacent segment instability is one of the reasons that induce neck symptoms.

Adult↗
PubMed2004

Recognition and cleavage of primary microRNA precursors by the nuclear processing enzyme Drosha.

A critical step during human microRNA maturation is the processing of the primary microRNA transcript by the nuclear RNaseIII enzyme Drosha to generate the approximately 60-nucleotide precursor microRNA hairpin. How Drosha recognizes primary RNA substrates and selects its cleavage sites has remained a mystery, especially given that the known targets for Drosha processing show no discernable sequence homology. Here, we show that human Drosha selectively cleaves RNA hairpins bearing a large (>/=10 nucleotides) terminal loop. From the junction of the loop and the adjacent stem, Drosha then cleaves approximately two helical RNA turns into the stem to produce the precursor microRNA. Beyond the precursor microRNA cleavage sites, approximately one helix turn of stem extension is also essential for efficient processing. While the sites of Drosha cleavage are determined largely by the distance from the terminal loop, variations in stem structure and sequence around the cleavage site can fine-tune the actual cleavage sites chosen.

Animals↗
PubMed2004

Statistical mechanics of base stacking and pairing in DNA melting.

We propose a statistical mechanics model for DNA melting in which base stacking and pairing are explicitly introduced as distinct degrees of freedom. Unlike previous approaches, this model describes thermal denaturation of DNA secondary structure in the whole experimentally accessible temperature range. Base pairing is described through a zipper model, base stacking through an Ising model. We present experimental data on the unstacking transition, obtained exploiting the observation that at moderately low pH this transition is moved down to experimentally accessible temperatures. These measurements confirm that the Ising model approach is indeed a good description of base stacking. On the other hand, comparison with the experiments points to the limitations of the simple zipper model description of base pairing.

Base Pairing↗
PubMed2004

Structural requirements for pre-microRNA binding and nuclear export by Exportin 5.

The biogenesis and function of mature human microRNAs is dependent on the nuclear export of pre-microRNA precursors by Exportin 5 (Exp5). The precursor for the human miR-30 microRNA, which is a 63 nt long RNA hairpin bearing a 2 nt 3' overhang, forms a specific complex with Exp5 and the Ran-GTP cofactor. Here, we have examined the structural requirements for pre-microRNA binding by Exp5. Our data indicate that pre-miR-30 binding requires an RNA stem of >16 bp and is facilitated by a 3' overhang. Although a blunt-ended derivative of the pre-miR-30 stem-loop remained capable of binding Exp5, 5' overhangs were inhibitory. miR-30 variants that had lost the ability to bind Exp5 effectively were not efficiently exported from the nucleus and were also expressed at reduced levels. Furthermore, formation of a pre-microRNA/Exp5/Ran-GTP complex inhibited exonucleolytic digestion of the pre-miRNA in vitro. Together, these data demonstrate that pre-microRNA binding by Exp5 involves recognition of almost all of the RNA hairpin, with the exception of the terminal loop. Moreover, these results argue that Exp5 binding not only mediates pre-microRNA nuclear export but also prevents nuclear pre-microRNA degradation.

Active Transport, Cell Nucleus↗
PubMed2004

Bubble nucleation and cooperativity in DNA melting.

Bubbles in DNA are related to fundamental processes such as duplication and transcription. Using a new ensemble technique to trap intermediate states, we present direct measurements of the average length of the denaturation bubble and the statistical weights of the bubble states in the temperature-driven melting of DNA oligomers. For a bubble flanked by double-stranded regions, we find a nucleation size of approximately 20 bases, and a broad distribution of bubble sizes. However, for bubbles opening at the ends of the molecule there is no nucleation threshold. The measured statistical weights of different conformations agree with the predictions of the thermodynamic models in the case of unzipping from the ends; however, internal bubble states are not completely described by the models. The measurements further show that, due to end effects, the melting transition becomes a two-state process only in the limit of a molecule length L approximately 1 bp.

Chemical Phenomena↗
PubMed2004

DNA minigene vaccination for adjuvant neuroblastoma therapy.

The disruption of self-tolerance against neuroblastoma is the ultimate goal of an effective DNA-vaccine. We demonstrate the induction of protective immunity against syngeneic murine NXS2 neuroblastoma in A/J mice following vaccination with tyrosine hydroxylase (TH)-derived antigens. Oral gene delivery was accomplished using an attenuated strain of Salmonella typhimurium as a carrier harboring vectors encoding for mouse tyrosine hydroxylase (mTH) antigens. Vaccination was effective in protecting animals from a lethal challenge with wild-type NXS2 tumor cells. These findings were extended by comparing efficacy of mTH minigene vaccines with a minigene vaccine comprising three novel epitopes isolated fom NXS2 neuroblastoma cells. For this purpose, MHC class I was immunoprecipitated from NXS2 cell lysates, and peptides were eluted and examined in tandem-mass spectrometry analysis. This led to the identification of three novel natural MHC class I peptide ligands: TEALPVKLI, from ribonucleotide reductase M2; NEYIMSLI, from Ser/Thr protein phosphatase 2A; and FEMVSTLI, of unknown origin. Two minigenes were constructed, one encoding for the three novel epitopes and the second for three known mTH-derived epitopes with high predicted binding affinity to MHC class I, by cloning them into the mammalian expression vector pCMV-3FUB. Immunized mice showed a reduction in primary tumor growth and the absence of spontaneous liver metastasis in the majority of animals. Importantly, there was no significant difference between the two minigenes, suggesting that, compared with tumor peptide isolation, mTH epitope prediction is similarly effective for designing efficient DNA-minigene vaccines. In summary, these findings establish proof of the concept that disruption of self-tolerance against neuroblastoma-associated epitopes may be an effective adjuvant therapeutic strategy.

Animals↗
PubMed2004

[A preliminary study using RNA interference technique against replication of HBV in vitro].

OBJECTIVE: To identify the siRNA interference ability for the replication of HBV. METHODS: Based on the sequence of HBV in HepG2 2.2.15 cells in GenBank, one sequence targeting the C antigen of HBV was cloned into the RNA polymerase III based expression vector pSuper. This recombinant was electroporated into HepG2 2.2.15 cells and the expression of HBsAg and HBeAg was assayed using ELISA. RESULTS: The construction of the recombinant expression vector pSuper-C and its control vector pSuper was successfully confirmed by the results of enzyme digestion, electrophoresis and sequencing. However, there was no difference between the expression of HBsAg and HBeAg in the supernatant of HepG2 2.2.15 cell culture in the experimental and control groups. CONCLUSIONS: The constructed pSuper-C did not show an interfering effect on the replication of HBV in HepG2 2.2.15 cells. In order to display this effect, further study is needed

Hepatitis B virus↗
PubMed2003

Length and statistical weight of bubbles in DNA melting.

We present measurements of the length of the bubble versus temperature in the melting transition of DNA oligomers. The sequences are clamped at the ends of GC pairs (strong binding), and have AT rich (weaker binding) middle regions of variable length B relative to the molecule size. We use a quenching technique to trap intermediate states. We find that the average relative bubble size grows from zero for increasing temperature, but reaches a plateau at a length of order B, producing a discontinuous transition with finite bubble size. The statistical weight of the bubble states decreases as B is reduced, the transition becoming more cooperative.

Base Sequence↗
PubMed2003

MicroRNAs and small interfering RNAs can inhibit mRNA expression by similar mechanisms.

MicroRNAs (miRNAs) are endogenously encoded small noncoding RNAs, derived by processing of short RNA hairpins, that can inhibit the translation of mRNAs bearing partially complementary target sequences. In contrast, small interfering RNAs (siRNAs), which are derived by processing of long double-stranded RNAs and are often of exogenous origin, degrade mRNAs bearing fully complementary sequences. Here, we demonstrate that an endogenously encoded human miRNA is able to cleave an mRNA bearing fully complementary target sites, whereas an exogenously supplied siRNA can inhibit the expression of an mRNA bearing partially complementary sequences without inducing detectable RNA cleavage. These data suggest that miRNAs and siRNAs can use similar mechanisms to repress mRNA expression and that the choice of mechanism may be largely or entirely determined by the degree of complementary of the RNA target.

Base Sequence↗
PubMed2003

Allyl isothiocyanate, a constituent of cruciferous vegetables, inhibits proliferation of human prostate cancer cells by causing G2/M arrest and inducing apoptosis.

Dietary isothiocyanates (ITCs) are highly effective in affording protection against chemically induced cancers in laboratory animals. In the present study, we demonstrate that allyl isothiocyanate (AITC), a constituent of cruciferous vegetables, significantly inhibits proliferation of cultured PC-3 (androgen-independent) and LNCaP (androgen-dependent) human prostate cancer cells in a dose-dependent manner with an IC(50) of approximately 15-17 micro M. On the other hand, survival of a normal prostate epithelial cell line (PrEC) was minimally affected by AITC even at concentrations that were highly cytotoxic to the prostate cancer cells. Reduced proliferation of PC-3 as well as LNCaP cells in the presence of AITC correlated with accumulation of cells in G(2)/M phase and induction of apoptosis. In contrast, AITC treatment failed to induce apoptosis or cause G(2)/M phase arrest in PrEC cells. A 24 h treatment of PC-3 and LNCaP cells with 20 micro M AITC caused a significant decrease in the levels of proteins that regulate G(2)/M progression, including Cdk1 (32-50% reduction), Cdc25B (44-48% reduction) and Cdc25C (>90% reduction). A significant reduction in the expression of cyclin B1 protein (approximately 45%) was observed only in LNCaP cells. A 24 h exposure of PC-3 and LNCaP cells to an apoptosis-inducing concentration of AITC (20 micro M) resulted in a significant decrease (31-68%) in the levels of anti-apoptotic protein Bcl-2 in both cell lines, and approximately 58% reduction in Bcl-X(L) protein expression in LNCaP cells. In conclusion, it seems reasonable to hypothesize that AITC, and possibly other ITCs, may find use in the treatment of human prostate cancers.

Apoptosis↗
PubMed2003

Photoperiodic regulation of a 24-kD dehydrin-like protein in red-osier dogwood (Cornus sericea L.) in relation to freeze-tolerance.

A predominant 24-kD dehydrin-like protein was previously found to fluctuate seasonally within red-osier dogwood (Cornus sericea L.) stems. The current study attempted to determine what environmental cues triggered the accumulation of the 24-kD protein and to assess its potential role in winter survival. Controlled photoperiod and field studies confirmed that photoperiod regulates a reduction of stem water content (SWC), freeze-tolerance enhancement and accumulation of the 24-kD protein. Diverse climatic ecotypes, which are known to respond to different critical photoperiods, displayed differential reduction of SWC and accumulation of the 24-kD protein. A time-course study confirmed that prolonged exposure to short days is essential for SWC reduction, 24-kD protein accumulation, and freeze-tolerance enhancement. Water deficit induced 24-kD protein accumulation and enhanced freeze-tolerance under long-day conditions. In all instances, freeze-tolerance enhancement and 24-kD protein accumulation was preceded by a reduction of SWC. These results are consistent with the hypothesis that C. sericea responds to decreasing photoperiod, which triggers a reduction in SWC. Reduced SWC in turn may trigger the accumulation of the 24-kD protein and a parallel increase in freeze-tolerance.

Acclimatization↗
PubMed2003

Identification of a retinoid receptor response element in the human aldehyde dehydrogenase-2 promoter.

BACKGROUND: The human aldehyde dehydrogenase-2 promoter contains sites that bind members of the nuclear receptor family, and one (designated FP330-3') is predicted to bind retinoic acid receptors. METHODS: Binding of retinoid receptors to the FP330-3' oligonucleotide duplex and point mutations thereof was assayed using electrophoretic mobility shift assays. The function of the promoter element was determined in transfection assays. RESULTS: Heterodimers of retinoic acid receptor (RAR)alpha, beta, and gamma with retinoid X receptor (RXR)alpha bound the FP330-3' site. Mutagenesis of the FP330-3' site suggested that either the upstream DR-5 or downstream DR-1 could mediate binding of RAR/RXR. FP330-3' oligonucleotide duplexes were not bound by in vitro translated RXR homodimers but weakly competed with a synthetic DR-1 oligonucleotide duplex for binding by RXR. A reporter construct carrying four copies of the FP330-3' element was induced by cotransfection of rat hepatoma cells with a construct encoding RARalpha, when the RAR-specific ligand AM580 was present. Each of the three RXR isoforms alpha, beta, and gamma stimulated the expression of reporter constructs containing the FP330-3' sites in a 9-cis retinoic acid-dependent fashion in cells in culture. This was confirmed in the case of RXRalpha using the RXR-specific ligand methoprene. CONCLUSION: The human aldehyde dehydrogenase-2 promoter contains a retinoid response element, which may contribute to regulation of the gene.

Aldehyde Dehydrogenase↗
PubMed2003

Sequence requirements for micro RNA processing and function in human cells.

Most eukaryotes encode a substantial number of small noncoding RNAs termed micro RNAs (miRNAs). Previously, we have demonstrated that miR-30, a 22-nucleotide human miRNA, can be processed from a longer transcript bearing the proposed miR-30 stem-loop precursor and can translationally inhibit an mRNA-bearing artificial target sites. We also demonstrated that the miR-30 precursor stem can be substituted with a heterologous stem, which can be processed to yield novel miRNAs and can block the expression of endogenous mRNAs. Here, we show that a second human miRNA, termed miR-21, can also be effectively expressed when its precursor forms part of a longer mRNA. For both miR-30 and miR-21, mature miRNA production was highly dependent on the integrity of the precursor RNA stem, although the underlying sequence had little effect. In contrast, the sequence of the terminal loop affected miRNA production only moderately. Processing of the initial, miR-30-containing transcript led to the production of not only mature miR-30 but also to the largely nuclear excision of an approximately 65-nucleotide RNA that is likely to represent an important intermediate in miR-30 processing. Consistent with this hypothesis, mutations that affected mature miR-30 production inhibited expression of this miR-30 pre-miRNA to an equivalent degree. Although point mutations could block the ability of both miR-30 and miR-21 to inhibit the translation of mRNAs bearing multiple artificial miRNA target sites, single point mutations only attenuated the miRNA-mediated inhibition of genes bearing single, fully complementary targets. These results suggest that miRNAs, and the closely similar small interfering RNAs, cannot totally discriminate between RNA targets differing by a single nucleotide.

Base Sequence↗
PubMed2003

[The primary study on the anti-HBV effect of whole recombinant yeast].

OBJECTIVES: Based on the immunologic character of Pichia pastoris yeast, a new therapeutic vaccine, whole recombinant yeast, was used to explore a new way to activate cell-mediated anti-viral immunity. METHODS: The recombinant plasmids, pPIC9K/S and PIC9K/hsp(1-370)-S, were constructed by inserting the gene encoding HBsAg, HSP70 (1-370) -HBsAg into vector pPIC9K and then the recombinants were transfected into Pichia pastoris yeast,GS115, respectively. Then that recombinant yeast immunized BALB/C mice were detected for humoral and cellular immunity to HBsAg. RESULTS: Recombinant yeast successfully activated the humoral immunity to HBsAg in mice, but failed to activate the cellular immunity. CONCLUSION: The whole recombinant yeast can be used as vaccine, but need further study for optimal way of immunization.

Animals↗
PubMed2003

[Archaeal diversity of Zabuye Lake in Tibet analyzed by culture-independent approach].

With culture-independent approach, microbial DNA was directly extracted from samples of Zabuye saline soda lake. Using the microbial DNA as template, archaeal 16S rDNAs were amplified by PCR. Amplified products were cloned and sequenced. 60 different cloned partial sequences, most of which were related to haloalkaliphilic archaeon, were acquired. In the phylogenetic tree, some clones of Zabuye lake belonged to Genus Natronobacterium, Natrinema, Natronococcus, Natronorubrum, Natronomonas, Halorubrum, Haloterrigena, Halorhabdus in Family Halobacteriaceae. Other clones represent some novel groups. All of them show prolific archaeal diversity of Zabuye lake.

Archaea↗
PubMed2002

Both natural and designed micro RNAs can inhibit the expression of cognate mRNAs when expressed in human cells.

Animal cells have recently been shown to express a range of approximately 22 nucleotide noncoding RNAs termed micro RNAs (miRNAs). Here, we show that the human mir-30 miRNA can be excised from irrelevant, endogenously transcribed mRNAs encompassing the predicted 71 nucleotide mir-30 precursor. Expression of the mir-30 miRNA specifically blocked the translation in human cells of an mRNA containing artificial mir-30 target sites. Similarly, designed miRNAs were also excised from transcripts encompassing artificial miRNA precursors and could inhibit the expression of mRNAs containing a complementary target site. These data indicate that novel miRNAs can be readily produced in vivo and can be designed to specifically inactivate the expression of selected target genes in human cells.

Animals↗