Biomedical subjects

Jin Zhao

Publications and source records attributed to Jin Zhao.

At least 37 records · Page 2Linked to original sources

PubMed2005

Intracellular co-localization of SPLUNC1 protein with nanobacteria in nasopharyngeal carcinoma epithelia HNE1 cells depended on the bactericidal permeability increasing protein domain.

Epithelial surfaces constitute natural immunobarriers against environmental threats. These barriers are brimming with fluids that bind, transport, cleave or degrade bacterial cells and their endotoxic by-products. Saliva and the airway surface-lining fluid (ASL) comprise the important fluid constituents. Short palate, lung and nasal epithelium clone 1 (SPLUNC1) is a potential host defensive protein that is secreted from the submucosal gland to the saliva and nasal lavage fluid. However, its antimicrobial spectrum and antimicrobial mechanism is not clear. Through green fluorescence protein (GFP) mediated subcellular localization experiments in nasopharyngeal carcinoma (NPC) HNE1 cell line, we determined that the intracellular GFP-tagged SPLUNC1 protein binds to a miniscule microorganisms, approximately 50-400nm in size, after the bactericidal permeability increasing protein (BPI) domain was deleted, GFP-tagged truncated SPLUNC1 protein lost its function of binding to the miniscule microorganisms. We verified that these microorganisms are nanobacteria (NB) with a negative staining using transmitted electronic microscope (TEM) and immunofluorescent analysis using an NB-specific antibody. We isolated and cultured the NB from the cultured nasopharyngeal carcinoma epithelia HNE1 cell supernatant. We found that the NB did not absorb the Hoechst stain, even when we extended the staining time to 35min. However, with the time extension the larger sized NB (larger than 300nm) did stain positively. From the biopsy specimen of NPC, we also detected the NB, which can lead to the swelling of mitochondria in the infected host cells. We hypothesize that SPLUNC1 and NB co-localization is due to the GFP-tagged SPLUNC1 protein binding to the lipopolysaccharide (LPS) of the Gram-negative NB, which can play an important role in the host defense of nasopharyngeal epithelium. This research sheds new light on the mechanism of SPLUNC1 involvement in the host upper respiratory tract defense system.

Antimicrobial Cationic Peptides↗

PubMed2005

Tissue distribution of the secretory protein, SPLUNC1, in the human fetus.

We previously identified a tissue-specific gene, short palate, lung, and nasal epithelium clone 1 (SPLUNC1), in nasopharyngeal epithelial tissues. SPLUNC1 was differentially expressed in nasopharyngeal carcinoma. Bioinformatic analysis revealed that SPLUNC1 has the bactericidal permeability-increasing protein/lipid-binding protein (BPI/LBP) domain and a 19 amino acid signal peptide, which suggest that it is a secretory protein. Its precise cellular localization in the respiratory tract is mainly in mucous cells and ducts of submucosal glands. However, little is known about its expression pattern in various human tissues. We generated a highly specific antibody and analyzed its distribution in the human fetus by immunohistochemistry to more precisely determine SPLUNC1 protein localization in human tissues. The results were further validated by RT-PCR. Our results showed that SPLUNC1 protein is expressed at not only the serous glands and epithelium of the upper respiratory tract and digestive tract, but also in the oculi of human embryos. Interestingly, we also found positive staining in fetus adipose tissue, a result not previously reported in studies of adult human tissues. Western blot analysis detected a 24 kDa SPLUNC1 protein in the compounds of nasopharyngeal secretions. This secretory protein was also detected in saliva and tears. Our research suggests that SPLUNC1 protein may not only be an antimicrobial peptide that plays an important role in the maintenance of homeostasis in the upper respiratory tract, oculi, and alimentary tract, it may also be important in the development and lipid metabolism of the adipose tissue.

Adipose Tissue↗

PubMed2005

Jade-1, a candidate renal tumor suppressor that promotes apoptosis.

Medical therapies are lacking for advanced renal cancer, so there is a great need to understand its pathogenesis. Most renal cancers have defects in the von Hippel-Lindau tumor suppressor pVHL. The mechanism by which pVHL protein functions in renal tumor suppression remains unclear. Jade-1 is a short-lived, kidney-enriched transcription factor that is stabilized by direct interaction with pVHL. Loss of Jade-1 stabilization by pVHL correlates with renal cancer risk, making the relationship between Jade-1 and renal cancer compelling. We report that Jade-1 expression was barely detectable in all tested renal cancer cell lines, regardless of VHL status. Strikingly, proteasome inhibitor treatment increased endogenous Jade-1 expression up to 10-fold. Jade-1 inhibited renal cancer cell growth, colony formation, and tumor formation in nude mice. Intriguingly, Jade-1 also affected the pattern of cell growth in monolayer culture and 3D culture. Jade-1 increased apoptosis by 40-50% and decreased levels of antiapoptotic Bcl-2. Antisense Jade-1-expressing cells confirmed these results. Therefore, Jade-1 may suppress renal cancer cell growth in part by increasing apoptosis. Jade-1 may represent a proapoptotic barrier to proliferation that must be overcome generally in renal cancer, perhaps initially by pVHL inactivation and subsequently by increased proteasomal activity. Therefore, Jade-1 may be a renal tumor suppressor.

Animals↗

PubMed2005

Single C59N molecule as a molecular rectifier.

We report a new kind of experimental realization of a molecular rectifier, which is based on a single azafullerene C59N molecule in a double-barrier tunnel junction via the single electron tunneling effect. An obvious rectifying effect is observed. The positive onset voltage is about 0.5-0.7 V, while the negative onset voltage is about 1.6-1.8 V. Theoretical analyses show that the half-occupied molecular orbital of the C59N molecule and the asymmetric shift of the molecular Fermi level when the molecule is charged are responsible for the molecular rectification.

Journal Article↗

PubMed2005

Adrenomedullin(27-52) inhibits vascular calcification in rats.

Adrenomedullin (ADM) has the vasodilatory properties and involves in the pathogenesis of vascular calcification. ADM could be degraded into more than six fragments in the body, including ADM(27-52), and we suppose the degrading fragments from ADM do the same bioactivities as derived peptides from pro-adrenomedullin. The present study carries forward by assessing the effects on vascular calcification of the systemic administration of ADM(27-52). The rat vascular calcific model was replicated with vitamin D3 and nicotine. ADM or/and ADM(27-52) were systemically administrated with mini-osmotic pump beginning at seventh day after the model replication for 25 days. Vascular calcific nodules histomorphometry, vascular calcium content, vascular calcium uptake, alkaline phosphatase activity, and osteopontin-mRNA quantification in aorta were assessed. ADM limited 40.2% vascular calcific nodules (P<0.01), did not effect on calcium content (P>0.05), reduced 44.4% calcium uptake (P<0.01), lowered 21.1% alkaline phosphatase activity (P<0.01), and regulated 40.9% downwards osteopontin-mRNA expression (P<0.01) in the aorta of rats with vascular calcification. ADM(27-52) receded 32.0% vascular calcific nodules (P<0.01), taken from 55.5% calcium content (P<0.01), did not affect calcium uptake (P>0.05), inhibited 22.5% alkaline phosphatase activity (P<0.01), and restrained 21.9% osteopontin-mRNA expression (P<0.01) in the aorta of rats with vascular calcification. Both of ADM and ADM(27-52) did interact on vascular calcification each other. ADM could partially antagonize the effects of ADM(27-52) in taking from calcium content (17.5%, P<0.01) and in receding vascular calcific nodules (18.6%, P<0.01). ADM could obviously enhance the action of ADM(27-52) in inhibiting alkaline phosphatase activity (14.4%, P<0.01) and in reducing calcium uptake (11.4%, P<0.01). ADM(27-52) could partially antagonize the effects of ADM on regulating downwards osteopontin-mRNA expression (17.0%, P<0.01). It is concluded that ADM(27-52) derived from ADM acts as an inhibitory agent on vascular calcification, with special mechanisms different from ADM derived from ADM progenitor molecule.

Adrenomedullin↗

PubMed2005

Wet electrons at the H2O/TiO2(110) surface.

At interfaces of metal oxide and water, partially hydrated or "wet-electron" states represent the lowest energy pathway for electron transfer. We studied the photoinduced electron transfer at the H2O/TiO2(110) interface by means of time-resolved two-photon photoemission spectroscopy and electronic structure theory. At approximately 1-monolayer coverage of water on partially hydroxylated TiO2 surfaces, we found an unoccupied electronic state 2.4 electron volts above the Fermi level. Density functional theory shows this to be a wet-electron state analogous to that reported in water clusters and which is distinct from hydrated electrons observed on water-covered metal surfaces. The decay of electrons from the wet-electron state to the conduction band of TiO2 occurs in </=15 femtoseconds.

Journal Article↗

PubMed2005

Substituted aminobenzimidazole pyrimidines as cyclin-dependent kinase inhibitors.

A series of aminobenzimidazole-substituted pyrimidines were synthesized and evaluated for biochemical activity against CDK1. A high-speed parallel synthesis approach enabled the identification of a potent lead series having improved potency in the CDK1 assay (IC(50)<10nM). Cell cycle analysis showed that the compounds induced a G2/M block. Docking studies were carried out with a CDK1 homology model, and provide a rationale for the observed activities.

Benzimidazoles↗

PubMed2005

Uniform distribution of epithelial stem cells in the bulbar conjunctiva.

PURPOSE: To determine the distribution of epithelial stem cells in the bulbar conjunctiva by measuring homeostatic movements and mitosis of epithelial cells in this region. METHODS: The ubiquitous GFP mouse was used to monitor movement of conjunctival epithelial cells. Cell movement was determined by histology, analyzing the shape and distribution of GFP cell clusters in flat wholemount specimens, and by in vivo time-lapse microscopy, tracking the movement of GFP-positive cells in the bulbar conjunctiva near the limbus. Mitoses were determined by labeling DNA of adult mice with bromodeoxyuridine (BrdU) for 3 days. Label-retaining cells (LRCs) were determined by a pulse label of newborn mice with BrdU, followed by a chase of 6 to 7 weeks. RESULTS: Similar to the corneal epithelium, only some of the conjunctival epithelial cells expressed a high level of GFP. Histology showed that GFP-positive cells existed as clusters of several to several dozen cells. No stripe pattern of GFP was observed in any part of the conjunctiva, suggesting that directed cell movement was rare or nonexistent. Time-lapse analyses revealed that none of the tracked GFP clusters exhibited a continuous and directed movement and that most GFP clusters were stationary for several weeks and much longer in some occasions. BrdU labeling showed that GFP-positive cells in this region were mitotically active. BrdU pulse-chase experiments demonstrated that LRCs were distributed uniformly in this region. CONCLUSIONS: Epithelial cells of the bulbar conjunctiva near the limbus are mitotically active and yet they are generally immobile in a lateral direction, indicating that these cells are self-sufficient. These results, combined with the uniform distribution of LRCs, suggest that epithelial stem cells are distributed uniformly in this area.

Animals↗

PubMed2005

Antioxidative and immunomodulatory role of melatonin, sodium selenite, N-acetyl-L-cysteine and quercetin on human umbilical blood.

We have previously reported on the DNA oxidative damage and oxygen stress in healthy term neonates. The aim of the present study was to investigate the antioxidative and immunomodulatory role of melatonin, sodium selenite, N-acetyl-L-cysteine and quercetin on umbilical blood. The single cell gel electrophoresis (comet) assay was used for DNA oxidative damage. The lymphocytes proliferation and natural killer (NK) activity in umbilical blood were assayed by 3[H]-thymidine uptaken and MTT methods. Results using comet assay showed protection by melatonin, N-acetyl-L-cysteine (NAC) and quercetin, and a protective and damaging effect by selenite sodium. Melatonin, sodium selenite, NAC and quercetin greatly promoted the lymphocytes proliferation to IL-2. NK activity of the umbilical blood was significantly increased by melatonin and sodium selenite, but was not affected by NAC and quercetin. These data suggest that antioxidants are able to protect umbilical blood mononuclear cells against oxygen stress and affect oxygen stress mediated immune function inhibition of umbilical blood. These results will supply new experimental data for using umbilical blood to treat diseases.

Acetylcysteine↗

PubMed2005

Oxidative stress in Graves' disease patients and antioxidant protection against lymphocytes DNA damage in vitro.

DNA damage to peripheral blood lymphocytes of patients with Graves' disease (GD) was studied in vitro before and after treatment with antioxidants, melatonin, quercetin, N-acetylcysteine (NAC) and vitamin C. DNA damage (comet %) was remarkably higher in patients (23.7 +/- 5.5%) than that in healthy persons (9.8 +/- 3.2%, p < 0.01). Plasma malondialdehyde (MDA) content (7.90 +/- 1.77 microM) of patients was significantly higher than that of healthy persons (4.71 +/- 1.19 microM, p < 0.01). Also, the plasma total antioxidant capacity (TAC) (7.53 +/- 1.35 U/ml) in GD patients was significantly lower than that in healthy persons (10.56 +/- 2.21 U/ml, p < 0.01). Negative correlations were observed between plasma TAC and DNA damage in lymphocytes (r = -0.599, p < 0.01), and between plasma TAC and MDA (r = -0.40, p < 0.05) in GD patients. After treatment with 100 microM melatonin, quercetin or NAC for 4 h in vitro, DNA damage in lymphocytes in GD patients declined significantly (from 23.8 +/- 4.4% to 14.4 +/- 4.0%, p < 0.001 for melatonin, from 23.4 +/- 4.7% to 18.1 +/- 4.3%, p < 0.01 for quercetin, from 23.7 +/- 4.0% to 18.7 +/- 5.7%, p < 0.05 for NAC), while there was little change with concentrations of 1-100 microM of vitamin C. However, 1000 microM vitamin C enhanced DNA damage significantly (from 23.8 +/- 2.3% to 30.3 +/- 3.9%, p < 0.05). Our results showed that oxidative stress existed in GD patients and the antioxidants melatonin, quercetin and NAC are beneficial for DNA damage in lymphocytes of GD patients in vitro.

Acetylcysteine↗

PubMed2005

[Detection of SARS-coronavirus in both human and animals by RT-PCR].

OBJECTIVE: To find the most suitable RT-PCR detection method for SARS-Coronavirus (SARS-CoV) detection in both human and animals, and to study the source of SARS virus by investigating the condition of virus carried by wild, domestic animals and animals sold in market. METHODS: 350 throat washes of confirmed, suspected and observed SARS cases were tested by TaqMan and molecular beacon fluorescence RT-PCR methods. 386 animals with 442 nasal-throat swabs, fecal swabs of animals and cell cultured were detected by TaqMan method and conventional RT-PCR method. RESULTS: 10 positive were detected from 41 SARS clinical confirmed cases (24.39%). The results of TaqMan and molecular beacon are basically coincident with the coincident rate of 88.89%. 18 cell cultures with CPE were detected and find 16 positive, among which, 14 were civet cats (87.5%). The detecting results of animal samples from Dongmen Market (Shenzhen) and other markets and farms in peripheral areas show that: the total positive rate of 4 kinds of wild animal (civet cat, raccoon dog, hog-badge and Chinese ferret-badge) is 39.02% , which has an significant difference (P < 0.01) compared with the positive rate of other animals as 0. The positive rate of PCR for nasal and fecal swabs from 109 major wild animals is 44.04%, but the positive rate of 145 wild live-animals is 0. CONCLUSION: TaqMan and molecular beacon PCR methods both can be used in SARS detection.The results could support the hypothesis that animals (especially civet cat) could carry SARS virus.

Animals↗

PubMed2005

[Modified molecular beacon-based dual real-time PCR for detection of SARS virus and its application].

OBJECTIVE: To develop the modified molecular beacon-based dual fluorescent PCR assays for detection of SARS virus. The assay was applied to the early clinic diagnosis & animal tracking. METHODS: On the basis of the obtained core sequence of open reading frame 1b of the coronavirus polymerase gene sequences, which was published in GenBank, using modified molecular beacon probe, artifical virus techinique and two different fragments amplification with different fluoresce, one set of primers and probe were designed. Then fluorescent PCR assays for specific and sensitive detection of the SARS virus was established, while the ELISA & the traditional method were used as control. 368 clinical specimens such as the throat swab, serum, feces, and urine from different cases, 52 cell cultures and 50 animal specimens were detected by the molecular beacon-based PCR. RESULTS: The sensitivity of real -time PCR was 10 - 100 copies/ml, there was no cross reaction to other respiratory viruses such as influenza virus etc. Of 368 specimens, 20 were positive by using molecular beacon-based fluorescence PCR. The positive rate of SARS case (10/47) were 21.27%, the positive rate of the throat swab of SARS cases (10/23) were 43.87% . Among 52 SARS cell cultures, 29 were positive. The positive rate of SARS cell cultures was 55.77% . Of 50 animal specimens, 23 were positive. The positive rate was 46%. Furthermore, SARS virus RNA was detected in feces and in serum during the acute phase. CONCLUSION: The molecular beacon-based PCR is sensitive and specific, it could be applied to the early diagnosis and animal tracking. This molecular beacon-based PCR kit is useful for the different units.

Animals↗

PubMed2005

[Expression of p53 and Ki-67 genes in epithelial dysplasia from old oral mucosa and clinical significance].

OBJECTIVE: To elucidate the relationship between the malignant transformation of oral epithelial dysplasia and the expression of p53 and Ki-67 proteins. METHODS: The expression of p53 and Ki-67 proteins from 60 cases of oral epithelial dysplasia (28 cases of mild epithelial dysplasia, 32 cases of severe epithelial dysplasia) and 30 normal oral mucosa tissue was determined by immunihistochemistry, and a 5-year follow-up study was made. RESULTS: In normal epithelium, Ki-67 expression was only observed in a small number of cells, and no expression of p53 was detected. In the cases of mild epithelial dysplasia, low levels of Ki-67 and p53 expression were detected. In the cases of severe epithelial dysplasia, the expression of p53 and Ki-67 protein was significantly higher than that in the normal oral mucosa and in the mild epithelial cases (P < 0.05). There was a correlation between the malignant transformation and the mutation of p53, Ki-67 in the cases of oral epithelial dysplasia. CONCLUSION: Carcinogenesis of oral mucosa is a continuous and stepwise course; p53 and Ki-67 genes may play an important role in this course.

Aged↗

PubMed2005

[Influences of MyoG gene on reproductive traits in Jinhua pig].

The genotypes in the second intron and 3' UTR region of myogenin gene in 149 Jinhua pigs (line I, 109; line II, 15; line III, 25) were detected by PCR-RFLP method. The results showed that three genotypes (AA, AB and BB) in the PCR1-MspI-RFLP site were detected and their frequencies were 0.1544, 0.3826, 0.4631, respectively; MM and MN genotypes were found in PCR2-MspI-RFLP site and their frequencies were 0.9953 and 0.0047. The effect of MyoG gene on the reproductive traits was analyzed using SPSS software. Significant associations were found between MyoG locus and total number born (TNB) in first parity (P<0.05), and number born alive (NBA) in the second parity (P<0.05), but no significant associations existed in other parity combinations of TNB, NBA and litter weight birth (LWB) (P<0.05).

3' Untranslated Regions↗

PubMed2005

[Determination of carbendazim and thiabendazole in tomatoes by solid-phase microextraction coupled with high performance liquid chromatography and fluorescence detection].

A novel method for the determination of carbendazim (MBC) and thiabendazole (TBZ) in tomatoes by solid-phase microextraction (SPME) coupled with high performance liquid chromatography (HPLC) and fluorescence detection was developed. The experimental conditions of SPME, including extraction fiber, extraction time, extraction temperature, desorption time, desorption solvent, desorption mode, pH value, organic solvent and ionic strength, and HPLC conditions were optimized. The SPME for MBC and TBZ was performed on a 65 microm polydimethylsiloxane/divinylbenzene (PDMS/DVB) fibre for 50 min at room temperature with the solution being stirred at 1 100 r/min. The florescence detection was made at 315 nm with excitation wavelength at 280 nm. The method is linear for MBC and TBZ over the range assayed from 0.01 to 1.0 mg/kg tomatoes with the detection limits of 0.003 mg/kg and 0. 001 mg/kg and the correlation coefficients of 0.995 8 and 0.996 7, respectively. The average recoveries for MBC and TBZ were 83.5% and 85.6% with the relative standard deviations (RSDs) of 6.5% and 3.8%, respectively. The method is fast, simple, sensitive, solvent-free and suitable for the determination of MBC and TBZ in tomatoes.

Benzimidazoles↗

PubMed2004

Heterogenisation of CpMo(CO)3Cl on mesoporous materials and its application as olefin epoxidation catalyst.

CpMo(CO)(3)Cl reacts with the hydroxyl (Si-OH or Si-OH-Al) functionalities of mesoporous molecular sieves such as MCM-41, MCM-48 and its aluminium analogues during grafting. XRD, N(2) adsorption-desorption, BET surface area analysis and TEM show the resulting samples as being well ordered and maintaining a uniform pore size. FT-IR spectra, elemental analysis, (13)C and (29)Si CP MAS NMR spectra confirm the successful grafting. In the presence of excess TBHP the materials show high activity in cyclooctene epoxidation and good stability.

Alkenes↗

PubMed2004

Identification of a novel box C/D snoRNA from mouse nucleolar cDNA library.

By construction and screen of mouse nucleolar cDNA library, a novel mammalian small nucleolar RNAs (snoRNA) was identified. The novel snoRNA, 70 nt in length, displays structural features typical of C/D box snoRNA family. The snoRNA possesses an 11-nt-long rRNA antisense element and is predicted to guide the 2'-O-methylation of mouse 28S rRNA at G4043, a site unknown so far to be modified in vertebrates. The comparison of functional element of snoRNA guides among eukaryotes reveals that the novel snoRNA is a mammalian counterpart of yeast snR38 despite highly divergent sequence between them. Mouse and human snR38 and other cognates in distant vertebrates were positively detected with slight length variability. As expected, the rRNA ribose-methylation site predicted by mouse snR38 was precisely mapped by specific-primer extension assay. Furthermore, our analyses show that mouse and human snR38 gene have multiple variants and are nested in the introns of different host genes with unknown function. Thus, snR38 is a phylogenetically conserved methylation guide but exhibits different genomic organization in eukaryotes.

Animals↗

PubMed2004

DNA damage in healthy term neonate.

BACKGROUND: The process of childbirth is accompanied by an increase in oxidative aggression. AIM: To determine DNA damage and oxidative stress in healthy term neonates at birth. DESIGN: A total of 34 healthy full-term neonates, 22 healthy adults and 20 samples of colostrum from mothers of full-term neonates were examined. The malondialdehyde (MDA), DNA damage, GSH/GSSG ratio and total antioxidant capacity (TAC) in mononuclear cells isolated from umbilical blood and adult peripheral blood were measured. Moreover, the TAC of colostrum was also measured. The protective activity of five natural polyphenols against H(2)O(2)-induced DNA damage in mononuclear cells of umbilical blood was studied. RESULTS: A high level of DNA damage (p<0.001) accompanied with lower TAC (p<0.05) and GSH/GSSG ratio (p<0.001) and with higher level of MDA (p<0.001) in umbilical blood compared with those of healthy adult peripheral blood. The natural polyphenols, 7,8-dihydroxy-4-methyl coumarin, quercetin and resveratrol, are able to protect mononuclear cells of umbilical blood from oxidative attack. However, other two polyphenols, rutin and 7-hydroxy-4-methyl coumarin, do not. The TAC of colostrum is significantly higher than that of umbilical blood (p<0.001). CONCLUSIONS: The DNA oxidative damage in mononuclear cells of umbilical blood as well as other indexes related to redox status provided evidence that a sudden increase in oxygenation exposes the neonate into oxidative stress. Colostrum with a significant high TAC is very important for health care in infants against the oxidative stress.

Antioxidants↗