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Biomedical subjects

H Mao

Publications and source records attributed to H Mao.

At least 55 records · Page 3Linked to original sources

Effects of chlorophyll availability on phycobilisomes in Synechocystis sp. PCC 6803.

Inactivation of the chlL gene in Synechocystis sp. PCC 6803 resulted in negligible chlorophyll content when the mutant was grown in darkness. Upon phycocyanin excitation at 580 nm, the 77K fluorescence spectrum of dark-grown cells showed three peaks at 648 nm, 665 nm, and 685 nm, this last being the largest. This reflects the functional presence of major components of phycobilisomes, including phycocyanin, allophycocyanin, and the terminal emitter, and efficient energy transfer between these components. As expected, no fluorescence emission peaks corresponding to chlorophyll in the photosystems were observed. Intact phycobilisomes could be isolated from the dark-grown chlL-deletion mutant. However, the phycobilisomes had a lower efficiency of energy transfer than did those isolated from the light-grown mutant, probably because of a decreased phycobilisome stability in the absence of chlorophyll. Exposing the dark-grown chlL-deletion mutant to light triggered the biosynthesis of chlorophyll. For the first 6 h in the light, upon phycocyanin excitation at 580 nm, the 77K fluorescence emission spectrum of greening cells was identical to that of dark-grown cells that lacked significant amounts of chlorophyll. With increased chlorophyll synthesis, gradual energy transfer from phycobilisomes to the two photosystems can be demonstrated.

Bacterial Proteins↗

Intrastrain variants of herpes simplex virus type 1 isolated from a neonate with fatal disseminated infection differ in the ICP34.5 gene, glycoprotein processing, and neuroinvasiveness.

Two intrastrain variants of herpes simplex virus type 1 (HSV-1) were isolated from a newborn with fatal disseminated infection. A small-plaque-producing variant (SP7) was the predominant virus (>99%) in the brain, and a large-plaque-producing variant (LP5) was the predominant virus (>99%) in the lung and gastrointestinal tract. EcoRI and BamHI restriction fragment patterns indicated that SP7 and LP5 are related strains. The large-plaque variants produced plaques similar in size to those produced by HSV-1 KOS. Unlike LP5 or KOS, SP7 was highly cell associated and processing of glycoprotein C and glycoprotein D was limited to precursor forms in infected Vero cells. The large-plaque phenotype from KOS could be transferred into SP7 by cotransfection of plasmids containing the EK or JK EcoRI fragment or a 3-kb plasmid with the UL34.5 gene of HSV-1 KOS together with SP7 DNA. PCR analysis using primers from within the ICP34.5 gene indicated differences for SP7, LP5, and KOS. Sequencing data indicated two sets of deletions in the UL34.5 gene that distinguish SP7 from LP5. Both SP7 and LP5 variants were neurovirulent (lethal following intracranial inoculation of young BALB/c mice); however, the LP5 variant was much less able to cause lethal neuroinvasive disease (footpad inoculation) whereas KOS caused no disease. Passage of SP7 selected for viruses (SLP-5 and SLP-10) which were attenuated for lethal neuroinvasive disease, were not cell-associated, and differed in the UL34.5 gene. UL34.5 from SLP-5 or SLP-10 resembled that of KOS. These findings support a role for UL34.5 in promoting virus egress and for neuroinvasive disease.

Amino Acid Sequence↗

[An experimental study of viscoelastic properties of articular-cartilage of patella].

OBJECTIVE: To study the viscoelastic properties of human articular cartilages of patella in 8 cadavers, which were obtained from patients ranged in age from nineteen to fifty-one years old and died of acute head injury. METHODS: The creep and stress relaxation of articular cartilage of patella were abserved in the condition of articular cartilage loaded of 500 Pa, just as the physiological load of articular cartilage of patella in keeping human body standing position with the knee joint flexed 30 degrees, during the experiment. The deformation-time curve under certain load, the load-time curve under certain deformity and related data were obtained. RESULTS: Creep and stress relaxation showed that obvious changes occured in the first 10-15 minutes during the whole process. CONCLUSIONS: The data and curve will not only be helpful to understand the pathogenesis of chondromalacia, but also propose a feasible method to study it from biomechanical view, meanwhile provide reference data for designing the prosthesis of patella and material choosing.

Adult↗

[Influence of icariin on cell membrane of highly metastatic human lung tumor cell line].

In this study, flow cytometry (FCM) and fluorescent probe technique were used to explore the influence of icariin (ICA) on cell membrane of PG. The data suggested that ICA could inhance membrane fluidity of PG and increase the expression of membrane HLA-ABC antigen. This could be one of the anticancer mechanisms of ICA.

Antineoplastic Agents, Phytogenic↗

In situ observations of a high-pressure phase of H2O Ice

A previously unknown solid phase of H2O has been identified by its peculiar growth patterns, distinct pressure-temperature melting relations, and vibrational Raman spectra. Morphologies of ice crystals and their pressure-temperature melting relations were directly observed in a hydrothermal diamond-anvil cell for H2O bulk densities between 1203 and 1257 kilograms per cubic meter at temperatures between -10 degrees and 50 degreesC. Under these conditions, four different ice forms were observed to melt: two stable phases, ice V and ice VI, and two metastable phases, ice IV and the new ice phase. The Raman spectra and crystal morphology are consistent with a disordered anisotropic structure with some similarities to ice VI.

Journal Article↗

An intercalated and thermally stable FAPY adduct of aflatoxin B1 in a DNA duplex: structural refinement from 1H NMR.

The structure of a formamidopyrimidine (FAPY) adduct arising from imidazole ring opening of the initially formed trans-8, 9-dihydro-8-(N7-guanyl)-9-hydroxyaflatoxin B1 adduct under basic conditions and positioned in the 5'-d(CTATFAPYGATTCA)-3'*5'-d(TGAATCATAG)-3' oligodeoxynucleotide was determined. The FAPY adduct may be a major progenitor of aflatoxin B1-induced mutations in DNA. The freshly prepared sample showed biphasic melting, with transitions at 28 and 56 degreesC. NMR initially showed multiple subspectra. Over a period of several days at 4 degreesC, the sample converted to a single species with a Tm of 56 degreesC, 15 degrees C greater than the unmodified duplex. The deoxyribose was in the beta configuration about the anomeric carbon, evidenced by NOEs between FAPYG5 H3', H2', H2", and H1'. FAPY formation resulted in the loss of the guanine H8 proton, and the introduction of the formyl proton, which showed NOEs to FAPYG5 H1' and A6 N6Ha. A total of 31 NOEs from AFB1 to DNA protons were observed, mostly to the 5'-neighboring base, T4 in the modified strand. Sequential NOEs were interrupted between T4 and FAPYG5 in the modified strand, between C16 and A17 in the complementary strand, and between T4 N3H and FAPYG5 N1H. An NOE between FAPYG5 N1H and C16 N4H showed intact hydrogen bonding at FAPYG5*C16. Upfield chemical shifts were observed for T4 H6 and A17 H8. Molecular dynamics calculations converged with pairwise rmsd differences of <0.9 A. The sixth root residual was 8.7 x 10(-2). The AFB1 moiety intercalated from the major groove between FAPYG5 and T4*A17, and stacked with T4 and FAPYG5 and partially stacked with A17. The base step between T4*A17 and FAPYG5*C16 was increased from 3.4 to 7 A. The duplex unwound by about 15 degrees. The FAPY formyl group was positioned to form a hydrogen bond with A6 N6Ha. Strong stacking involving the AFB1 moiety, and this hydrogen bond explains the thermal stabilization of four base pairs by this adduct, and may be a significant factor in its processing.

Aflatoxin B1↗

DNA binding properties of the ecdysteroid receptor in the salivary gland of the female ixodid tick, Amblyomma hebraeum.

Salivary gland degeneration in the female tick, Amblyomma hebraeum Koch (Acari: Ixodidae) is controlled by an ecdysteroid hormone. In an earlier study (Mao, H., McBlain, W.A., Kaufman, W.R., 1995. Some properties of the ecdysteroid receptor in the salivary gland of the ixodid tick, Amblyomma hebraeum. Gen. Comp. Endocrinol. 99, 340-348), we demonstrated that a protein component of a salivary gland extract binds to ponasterone A (Pon A) with high affinity (Kd-1 nM), suggesting a tick ecdysteroid receptor (EcR). In this study, the Pon A binding protein bound to calf thymus DNA; this binding could be dissociated by Drosophila hsp27 EcRE. The binding protein shifted the [32P]hsp27 EcRE band on a gel mobility shift assay; formation of the complex with hsp27 EcRE required KCl (optimal concentration was approximately 75 mM). A number of physiologically effective ecdysteroids enhanced the binding with the following order of potency: Pon A > Mur A > Mak A > 20E > ecdysone, whereas vertebrate steroids (estradiol, cholesterol, corticosterone, progesterone, testosterone) had no such effect. Using monoclonal antibodies against Drosophila EcR and USP, we found that AG 10.2 recognized three bands (90.5, 87.3 and 84 kDa for EcR) and AB11 recognized at least two major bands (50.3 and 47.1 kDa for USP) in the salivary gland extract by western blot analysis. In addition, AB11 supershifted the tick EcR-hsp27 EcRE band on a gel mobility shift assay, indicating that the tick EcR heterodimerized with a USP-like protein for DNA binding. Furthermore, selective mutations to the 15-basepair palindrome of hsp27 EcRE at positions-5, + 2, or adding a base to the spacer, resulted in considerably reduced affinity to the tick EcR/USP. We thus propose a sequence similarity of EcREs between A. hebraeum and its insect counterpart.

Animals↗

[Detection of urogenital malignant cells in the peripheral blood by nested RT-PCR using keratin 19 mRNA].

INTRODUCTION AND OBJECTIVES: It is reported that cytokeratin 19 (CK-19) mRNA is not expressed in the peripheral blood cells of the healthy subjects (Am. J. Pathol. 142: 1111. 1993). Detection of DNA fragments of CK-19 in the peripheral blood suggests the existence of epithelial malignant tumor cells. In this study, we detected CK-19 genes in peripheral blood of patients with urogenital malignancy, and thus clarified the possibility of understanding tumor expansion. METHODS: Mononuclear cells were separated from the peripheral blood of 39 patients with urogenital tumor and 9 controls. Total cellular RNA was extracted according to the method described elsewhere. The CK-19 gene expression was investigated using nested reverse transcription polymerase chain reaction (nested RT-PCR) and confirmed by Southern Blotting. These results were compared to the clinical stages. RESULTS: CK-19 mRNA was not detected in any of 9 controls. CK-19 mRNA was detected in 3 out of the 10 bladder cancer cases. Two had metastases and 1 had muscle invasion without metastasis. One of 6 urothelial cancer case of the upper urinary tract showed positive amplification, and had liver, bone and lymphnode metastases. After one course of chemotherapy, mRNA became negative. In prostate cancer cases, three among 7 were positive and all 3 cases had distant metastases. In renal, testicular and penile cancer cases, the positive ratio were 3/6, 1/8 and 2/2, respectively. All positive cases of renal and testicular cancer had distant metastases, but one case of penile cancer had no metastasis. CONCLUSION: The detection of CK-19 mRNA in the circulating blood by nested RT-PCR makes it possible to detect micrometastasis, to evaluate therapeutic effects and to predict the prognosis.

Adult↗

[Detection of PSA mRNA from the peripheral blood and pelvic lymph nodes in patients with prostatic cancer by means of reverse transcription-polymerase chain reaction (RT-PCR)].

BACKGROUND AND METHODS: To detect prostate cancer cells in the blood circulation and in the lymph nodes by RT-PCR methods, we examined two kinds of prostate specific antigens (PSA) primers and one prostate specific membrane antigen (PSM). PSa primer 1 was established by us, PSA primer 2 by Moreno et al and PSM primer by Israeli et al. RESULTS: Both PSA primers were specific for expression of PSA mRNA because in 12 kinds of urogenital culture cells only LNCaP cells, which produce PSA, expressed PSA mRNA by RT-PCR, PSA 1 was more sensitive than PSA 2 for detection of PSA mRNA in the circulating cells since PSA mRNA was detected in the blood circulating cells in 5 cases of stage D2 prostate cancer using PSA primer 1 but in only one was using PSA primer 2. PSM mRNA was detected in all 12 types of urogenital cancer cells and in the blood circulating cells not of prostate cancer patients but also of renal, bladder, testicular cancer patients and normal volunteers. PSA 1 was used to detect PSA mRNA from the samples of fine needle aspiration biopsy (FNAB) of pelvic lymph node, and PSA mRNA was positive in 10 FNAB samples including not only all 6 cytologically positive and two cytologically class III cases but also 2 of 8 cytologically negative cases. RT-PCR for FNAB samples of all 15 cases of bladder cancer were negative for the detection of PSA mRNA. CONCLUSION: Detection of PSA mRNA by RT-PCR in FNAB samples may be useful to diagnose pelvic lymph node metastasis and to furnish additional information for the cytological diagnosis of prostate cancer.

Aged↗

[The expression study of human stem cell factor (hSCF) in E. coli].

OBJECTIVE: To improve the expression level of recombinant human stem cell factor (rhSCF). METHODS: With PCR techniques and synthesized oligonucleotide as primers, the number of nucleotides between the SD sequence and the starting codon ATG of human stem cell factor cDNA was altered, and the preferable codons of E. coli in the N-terminal amino acid sequence were selected. RESULTS: SDS-PAGE electrophoresis indicated that the ratio of expressed rhSCF to total E. coli proteins increased from 12% to 40% by thermal inducing. With DNA sequencing, the reading frame of the gene was proved to be correct. The sequence of 17 N-terminal amino acids of purified recombinant hSCF has been proved to be identical to the natural hSCF, except the first amino acid-Met. CONCLUSIONS: These studies suggest that the 5'-flanking region of hSCF cDNA plays an important role in its gene expression.

Amino Acid Sequence↗

[Development and clinical significance of SEA-1 emergency micro-ventilator].

To improve the work of emergency care and early cardio-pulmonary-brin resuscitation in our county, which is characteristic of abruptness and contingency, a new type of ventilator is highly required. It is hoped that such a ventilafor has a simple safe valve and does not need electric current from the mains. Also, the ventilator should be small in size, light in weight, simple in operation, easy to carry, swift to assembly, and reliable in ventilation. With the principle of pneumatic logic elements and the design of the integration of pneumatic circuits, we have successfully developed the Emergency Micro-ventilator, which accords with the above requirements. It has been confirmed that the ventilator is very effectual and reliable in ventilation support for the patients(n = 55) under general anesthesia without any case of hypoxemia, hypercapnia, hypotension, arrhythmia and so on, and the mechanical performances are stable.

Equipment Design↗

Multivariable dependence of Fe-Mg partitioning in the lower mantle

High-pressure diamond-cell experiments indicate that the iron-magnesium partitioning between (Fe,Mg)SiO3-perovskite and magnesiowustite in Earth's lower mantle depends on the pressure, temperature, bulk iron/magnesium ratio, and ferric iron content. The perovskite stability field expands with increasing pressure and temperature. The ferric iron component preferentially dissolves in perovskite and raises the apparent total iron content but had little effect on the partitioning of the ferrous iron. The ferrous iron depletes in perovskite at the top of the lower mantle and gradually increases at greater depth. These changes in iron-magnesium composition should affect geochemical and geophysical properties of the deep interior.

Journal Article↗

Internal iliac arterial infusion chemotherapy for rabbit invasive bladder cancer.

BACKGROUND: Internal iliac arterial infusion (IA) chemotherapy has been used clinically for locally invasive bladder cancer, but there have been no experimental studies to actually demonstrate whether IA is more effective than intravenous infusion (i.v.) chemotherapy in this setting. METHODS: We compared the effects of IA and i.v. using a rabbit invasive bladder cancer model. A 0.2 mL suspension containing 2 x 10(6) VX2 cancer cells was inoculated into the posterior submucosa of the bladder. Two weeks later the rabbits were divided into 3 treatment groups of 8 rabbits each: controls, a group treated with IA consisting of 10 mg/kg carboplatin and 1 mg/kg pirarubicin once a week for 3 weeks (days 14, 21, and 28), and the third treated with the same regimen intravenously. RESULTS: All bladder tumors of the rabbits in the IA group decreased in size, and 3 of the tumors totally disappeared (37.5%). There was also no evidence of lung metastasis. All tumors in the rabbits in the i.v. treatment group increased in size (tumor volume of IA vs. i.v., P = 0.008) and 2 rabbits had lung metastases. All tumors of the control group increased in size and all rabbits had lung metastases. The concentrations of platinum and pirarubicin in the bladder tumors were significantly higher in the IA treatment group than those in the i.v. treatment group at time points from 5 to 10 minutes (P < 0.05) after drug infusion. CONCLUSION: The antitumor effect of IA may be due to higher drug concentrations in the early stage after drug delivery, and the initial circulation of high concentrations of drugs may be the most important factor in suppressing tumor growth.

Animals↗

Improved large scale culture of Methylophilus methylotrophus for 13C/15N labeling and random fractional deuteration of ribonucleotides.

Isotopic labeling of RNA with 13C and 15N has become a routine procedure in structural studies by NMR spectroscopy. The methodology in this paper describes the random fractional deuteration of RNA using the obligate methylotropic bacterium, Methylophilus methylotrophus. This bacterium was grown using a non-deuterated carbon source in 52:48 D20/H20 and we have shown that all protons in the ribonucleotides except for the ribose H1 become 52% randomly fractionally deuterated. Improved growth conditions for this organism are also described that yield higher cell densities in liquid culture, which is applicable for all labeling procedures.

Carbon Isotopes↗

Alpha helix-RNA major groove recognition in an HIV-1 rev peptide-RRE RNA complex.

The solution structure of a human immunodeficiency virus type-1 (HIV-1) Rev peptide bound to stem-loop IIB of the Rev response element (RRE) RNA was solved by nuclear magnetic resonance spectroscopy. The Rev peptide has an alpha-helical conformation and binds in the major groove of the RNA near a purine-rich internal loop. Several arginine side chains make base-specific contacts, and an asparagine residue contacts a G.A base pair. The phosphate backbone adjacent to a G.G base pair adopts an unusual structure that allows the peptide to access a widened major groove. The structure formed by the two purine-purine base pairs of the RRE creates a distinctive binding pocket that the peptide can use for specific recognition.

Amino Acid Sequence↗

Impact on Salivary Gland Degeneration by Putative Ecdysteroid Antagonists and Agonists in the Ixodid Tick Amblyomma hebraeum

Ecdysteroids cause salivary gland degeneration in female ixodid ticks. We tested the effects of the following compounds on salivary gland degeneration in the ixodid tick Amblyomma hebraeum Koch: HHCS (22S,23S-homocastasterone), SSBR (22S,23S-homobrassinolide), STGM (2alpha,3alpha(OH)2-Delta22-stigmasten-6-one), RH 5849, and RH 5992. The first three are brassinosteroids (putative ecdysone antagonists) and the last two are nonsteroidal mimics of ecdysone in a variety of insects. In vitro, HHCS (up to 4 &mu;g/ml, 8.4 &mu;M) did not attenuate degeneration caused by 20-hydroxyecdysone; on the contrary, it enhanced the degree of salivary gland degeneration. SSBR (up to 4.5 &mu;g/ml, 9 &mu;M) likewise did not reduce 20-hydroxyecdysone mediated degeneration. RH 5849 up to 15 &mu;g/ml (51 &mu;M) and RH 5992 up to 10 &mu;g/ml (28 &mu;M) had no ecdysone-mimicking effect. In vivo, both RH compounds had an ecdysone-mimicking effect. RH 5849 (but not RH 5992) at 10 &mu;g/tick increased ovary wet weight slightly. None of the brassinosteroids displaced a significant amount of [3H]ponasterone A (PoA) from the ecdysone receptor at 6 &mu;g/ml (12-14 &mu;M) or below. RH 5849 (14 &mu;g/ml, 47 &mu;M) displaced 8% of (PoA) binding; at 68 &mu;g/ml (230 &mu;M) displacement was 30%. For RH 5992 inhibition of PoA binding was 16, 33, and 43% at 4.5, 16, and 79 &mu;g/ml (13, 45, and 79 &mu;M), respectively. Overall, the brassinosteroids and the RH compounds do not act on the tick salivary gland ecdysteroid system in a way similar to the way they act on some insect systems.

Journal Article↗