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Biomedical subjects

L Cai

Publications and source records attributed to L Cai.

At least 109 records · Page 6Linked to original sources

[Interleukin-6 gene cloning, expression and purification].

OBJECTIVE: The main purpose of this paper is to study the batch production of recombinant human interleukin-6(rhIL-6). METHODS: The cloned rhIL-6 gene is under the control of T7 promoter of pET30a vector and expressed in E. coli. RESULTS: The ratio of expressed recombinant protein to total cell protein is more than 50%. The rhIL-6 molecular weight is 21,000, isoelectric point is 6.7. The purity of the rhIL-6 is more than 95%, and the activity of rhIL-6, determined by IL-6 dependent mice hybridoma cell line 7TD1 and MTT assay, is 0.35 ng/ml. CONCLUSIONS: All results mentioned show that rhIL-6 meets the request of the middle scale production.

Cloning, Molecular↗

[Effect of Yangxue bushen tablet on ovarian function in animal model of Yang deficiency].

OBJECTIVE: To study the therapeutic mechanism of Yangxue Bushen tablet (tablet for nourishing blood and tonifying Kidney) on the Kidney deficiency type of functional amenorrhea and infrequent menstruation. METHODS: Applied with orimeten, models of Yang deficiency in female rabbits were made, ikaclomine was taken as the control drug. Group I was the group of animal model of Yang deficiency; Group II was the ikaclomine treatment group; Group III was the high dosage treatment group; Group IV was the low dosage group; Group V normal rabbit treated with distilled water. The functional effect of gonadal axis was evaluated by the changes of the signs of animal histomorphology of ovary and uterus, and the level of beta-endorphin. RESULTS: The animals of Yang deficiency in Group II, III and IV recovered as a whole, and the changes of different levels of folliculi, morphosis of endometria and deciduous vaginal epithelial cell in group II, III and IV were more obviously than those in group I and V (P < 0.05). The content of beta-endorphin in blood plasma in group I was 106.6 pg/ml, which was lower than those in the other groups (P < 0.05), and the contents of beta-endorphin in blood plasma, in group III, IV and II were similar to that in group V (P < 0.05). CONCLUSION: Yangxue Bushen tablet has the effect on regulating menstruation by regulating the function of gonadal axis, regulating and promoting the ovarian function.

Amenorrhea↗

[Studies on extensive application of bromoacetamide in various types of schistosomiasis endemic area].

AIM: To provide scientific basis for the popularization and application of molluscide bromoacetamide. METHODS: Large scale application of bromoacetamide in twenty eight counties of four provinces was investigated and its molluscicidal effect in marshland and influence on marshland grass (Cyperus) and fishes were observed. RESULTS: High molluscicidal effect of bromoacetamide was found against both snails and snail eggs in various environments. According to the results observed in different fields, the applicable dosage of bromoacetamide is 1.5-2 g/m2 for spraying method, 1.5-2 g/m3 for immersing method and 1.5-2 g/m for immersing in combination with shovelling the turf along the bank. When it was applied in fish-raising ponds or rivers at an applicable dosage, no toxic effect on fishes was found. The spraying dosage of bromoacetamide did not affect the rice seedling but it did some damage to the leaf-tips of marshland grass, however, the grass did turn green and grow. CONCLUSION: Bromoacetamide has proved to be a safe and effective mulluscicde against Oncomelania under various field conditions.

Acetamides↗

[Evaluation of the effect of causes of death of advanced schistosomiasis patients on the life span of population by using potential years of life lost analysis].

AIM: To evaluate the effect of the causes of death of advanced schistosomiasis patients on the life span of population in an area where schistosomiasis had been under control by applying the indicators of potential years of life lost (PYLL) and rate of PYLL (PYLLR). METHODS: Using the formula of calculation of PYLL and PYLLR, the causes of death of 487 patients of advanced schistosomiasis in 3 previously endemic townships of Shanghai suburb. RESULTS: Of 487 cases who died between 1955 and 1995, the PYLLR of all causes of death was 256.3@1000. The main causes of death were hepatic failure (100.3@1000), liver carcinoma (43.4@1000) and cancer of other organs (24.8@1000). In the 1960s, the PYLLR of advanced schistosomiasis complicated with hepatic failure and upper digestive tract hemorrhage was 222.2@1000 and 34.5@1000, the PYLLR of the complications of schistosomal liver cirrhosis, hepatic failure and hemorrhage accounted for 67.5% of the total PYLLR, being apparently higher in the non-splenectomy group than in the splenctomty group. After the schistosomiasis was under control, both PYLL and PYLLR decreased linearly. As compared with the 1960s, the PYLLR of all causes of death and the causes of death of the complications of schistosomal liver cirrhosis in the 1990s has declined by 62.9% and 83.2%, respectively, the PYLLR of upper digestive tract hemorrhage was found only 0.3@1000 in the last 5 years. CONCLUSION: PYLL and PYLLR analysis could be used to quantify the effect of complications of advanced schistosomiasis and other causes of death on the life span of population.

Adolescent↗

[The calculation of transitions 4s(2)4p-4s4p(2), 4s(2)4p-4s(2)5s, 4s(2)4p-4s(2)4d, 4s4p(2)-4p(3) and 4p(3)-4s(2)5s energy levels, wavelengths and oscillator strengthes in Ga-like ions from Z X-Rh XV].

In this paper, the 4s(2)4p, 4s(2)4d, 4s4p(2), 4p(3) and 4s(2)5s configuration's energy level structures, and configuration interactions in Ga-Like from Ga I-Xe XXIV ions are analysed in theory. The law of isoelectronic series is given. By using our fitted formula, the energies of Tc XIII ion 4s(2)4p, 4s4p(2) and 4s(2)5s configurations are predicted by interpolation, the energies of Tc IIIX-Rh XV ions 4p(3) and 4s(2)4d configurations are predicted by extrapolation. Wavelengths and oscillator strengthes (HXR theoretical calculation) of transitions 4s(2)4p-4s(2)4d, 4s(2)4p-4s4p(2), 4s(2)4p-4s(2)5s, 4p(3)-4s(2)5s and 4s(2)4p-4p(3) are given. The deviations of predictive energy levels from ob served values are less than 100cm(-1).

English Abstract↗

Effects of gamma radiation on levels of brain metallothionein and lipid peroxidation in transgenic mice.

The induction of metallothionein (MT) synthesis in the brain was investigated in MT-I isoform-overexpressing transgenic (MT-I*) and control mice after exposure to increasing doses of 2 to 20 Gy of whole-body gamma radiation. Although the MT-I isoform was the major isoform of MT in this transgenic mouse, the other isoforms, MT-II and MT-III, were also present in the brain. The total concentration of MT in the brain was measured by a cadmium-binding assay, while zinc and lipid peroxides were measured by atomic absorption spectrophotometry and by the thiobarbituric acid method, respectively. In MT-I* mice at 24 h after radiation exposure, the level of MT in the brain was increased from a basal level of 44.4 +/- 4.0 microg/g to a maximum level of 91.0 +/- 9.0 microg/g after 5 Gy and remained high after 10 and 20 Gy. In a time-course experiment with 5 Gy, the concentration of MT in the brain of MT-I* mice increased at 3 h and reached a maximum of 175.3 +/- 15.3 microg/g at 6 h. This high level of MT remained unchanged for 48 h after radiation exposure. Metallothionein was not induced markedly in the brains of control mice either at 24 h after exposure to 2-20 Gy gamma radiation or at different times after exposure to 5 Gy gamma radiation. In both strains of mice, the total concentration of zinc in the brain decreased with increasing radiation dose. No differences in lipid peroxide levels were seen in control mice exposed to 5 Gy at 6 and 12 h or after exposure to three other doses (2, 10 and 20 Gy) at 24 h. Slight increases (1.35 and 1.22, respectively) in lipid peroxide levels were observed in control mice at 24 and 48 h after exposure to 5 Gy. Lipid peroxide levels in the brain were not changed in irradiated MT-I* mice. The results show a marked increase in the levels of MT in the brain of transgenic mice after exposure to gamma radiation. The induced synthesis of MT may be only one of several mechanisms that prevent the induction of lipid peroxidation in the brain by gamma radiation.

Animals↗

Chemical oxidation and DNA damage catalysed by inorganic sunscreen ingredients.

Titanium dioxide (TiO2) has been noted (US Federal Register, 43FR38206, 25 August 1978) to be a safe physical sunscreen because it reflects and scatters UVB and UVA in sunlight. However, TiO2 absorbs about 70% of incident UV, and in aqueous environments this leads to the generation of hydroxyl radicals which can initiate oxidations. Using chemical methods, we show that all sunscreen TiO2 samples tested catalyse the photo-oxidation of a representative organic substrate (phenol). We also show that sunlight-illuminated TiO2 catalyses DNA damage both in vitro and in human cells. These results may be relevant to the overall effects of sunscreens.

Catalase↗

Tissue- and development-specific expression of multiple alternatively spliced transcripts of rat neuronal nitric oxide synthase.

Nitric oxide (NO) functions as an intercellular messenger and mediates numerous biological functions. Among the three isoforms of NO synthase that produce NO, the ubiquitously expressed neuronal NO synthase (nNOS) is responsible for a large part of NO production, yet its regulation is poorly understood. Recent reports of two alternative spliceforms of nNOS in the mouse and in man have raised the possibility of spatial and temporal modulation of expression. This study demonstrates the existence of at least three transcripts of the rat nNOS gene designated nNOSa, nNOSb, and nNOSc, respectively, with distinct 5' untranslated first exons that arise from alternative splicing to a common second exon. Expression of the alternative transcripts occurs with a high degree of tissue and developmental specificity, as demonstrated by RNase protection assays on multiple tissues from both fetal and adult rats. Furthermore, terminal differentiation of rat pheochromocytoma-derived PC12 cells into neurons is associated with induction of nNOSa, suggesting, likewise, development- and tissue-specific transcriptional control of nNOS isoform expression. Physical mapping using a rat yeast artificial chromosome clone shows that the alternatively spliced first exons 1a, 1b, and 1c are separated by at least 15-60 kb from the downstream coding sequence, with exons 1b and 1c being positioned within 200 bp of each other. These findings provide evidence that the biological activity of nNOS is tightly and specifically regulated by a complex pattern of alternative splicing, indicating that the notion of constitutive expression of this isoform needs to be revised.

Alternative Splicing↗

Local homogeneity of cell cycle length in developing mouse cortex.

We have measured the amount of variation in the length of the cell cycle for cells in the pseudostratified ventricular epithelium (PVE) of the developing cortex of mice on embryonic day 14. Our measurements were made in three cortical regions (i.e., the neocortex, archicortex, and periarchicortex) using three different methods: the cumulative labeling method (CLM), the percent labeled mitoses (PLM) method, and a comparison of the time needed for the PLM to ascend from 0 to 100% with the time needed for the PLM to descend from 100 to 0%. These 3 different techniques provide different perspectives on the cytokinetic parameters. Theoretically, CLM gives an estimate for a maximum value of the total length of the cell cycle (TC), whereas PLM gives an estimate of a minimum value of TC. The difference between these two estimates indicates that the range for TC is +/-1% of the mean TC for periarchicortex, +/-7% for neocortex, and +/-8% for archicortex. This was confirmed by a lengthening of the PLM descent time in comparison with its ascent time. The sharpness of the transitions and the flatness of the plateau of the PLM curves indicate that 99% of the proliferating cells are within this narrow estimated range for TC; hence, only approximately 1% deviate outside of a relatively restricted range from the average TC of the population. In the context of the possible existence within the cortical PVE of two populations with markedly dissimilar cell cycle kinetics from the mean, one such population must comprise approximately 99% of the total population, and the other, if it exists, is only approximately 1% of the total. This seems to be true for all three cortical regions. The narrow range of TC indicates a homogeneity in the cell cycle length for proliferating cells in three different cortical regions, despite the fact that progenitor cells of different lineages may be present. It further predicts the existence of almost synchronous interkinetic nuclear movements of the proliferating cells in the ventricular zone during early development of the cerebral cortex.

Animals↗

Synchrony of clonal cell proliferation and contiguity of clonally related cells: production of mosaicism in the ventricular zone of developing mouse neocortex.

We have analyzed clonal cell proliferation in the ventricular zone (VZ) of the early developing mouse neocortex with a replication-incompetent retrovirus encoding human placental alkaline phosphatase (AP). The retrovirus was injected into the lateral ventricles on embryonic day 11 (E11), i.e., at the onset of neuronogenesis. Three days postinjection, on E14, a total of 259 AP-labeled clones of various sizes were found in 7 fetal brains. There are approximately 7 cell cycles between E11 and E14 (), and there is a 1-2 cell cycle delay between retroviral injection and the production of a retrovirally labeled "founder" cell; thus, we estimate that the "age" of the clones was about 5-6 cell cycles. Almost one-half of the clones (48.3%) identified were pure proliferating clones containing cells only in the VZ. Another 18.5% contained both proliferating and postproliferative cells, and 33.2% contained only postproliferative cells. It was striking that over 90% of the clonally related proliferating cells occurred in clusters of two or more apparently contiguous cells, and about 73% of the proliferating cells occurred in clusters of three or more cells. Regardless of the number of cells in the clone, these clusters were tightly packed and confined to a single level of the VZ. This clustering of proliferating cells indicates that clonally related cells maintain neighbor-neighbor relationships as they undergo interkinetic nuclear migration and progress through several cell cycles, and, as a result, the ventricular zone is a mosaic of small clusters of clonally related and synchronously cycling cells. In addition, cells in the intermediate zone and the cortical plate were also frequently clustered, indicating that they became postproliferative at a similar time and that the output of the VZ is influenced by its mosaic structure.

Animals↗

Construction and characterization of a 10-genome equivalent yeast artificial chromosome library for the laboratory rat, Rattus norvegicus.

Increasing attention has been focused in recent years on the rat as a model organism for genetic studies, in particular for the investigation of complex traits, but progress has been limited by the lack of availability of large-insert genomic libraries. Here, we report the construction and characterization of an arrayed yeast artificial chromosome (YAC) library for the rat genome containing approximately 40,000 clones in the AB1380 host using the pCGS966 vector. An average size of 736 kb was estimated from 166 randomly chosen clones; thus the library provides 10-fold coverage of the genome, with a 99.99% probability of containing a unique sequence. Eight of 39 YACs analyzed by fluorescence in situ hybridization were found to be chimeric, indicating a proportion of about 20-30% of chimeric clones. The library was spotted on high-density filters to allow the identification of YAC clones by hybridization and was pooled using a 3-dimensional scheme for screening by PCR. Among 48 probes used to screen the library, an average of 9.3 positive clones were found, consistent with the calculated 10-fold genomic coverage of the library. This YAC library represents the first large-insert genomic library for the rat. It will be made available to the research community at large as an important new resource for complex genome analysis in this species.

Animals↗

Cancer chemopreventive activity of resveratrol, a natural product derived from grapes.

Resveratrol, a phytoalexin found in grapes and other food products, was purified and shown to have cancer chemopreventive activity in assays representing three major stages of carcinogenesis. Resveratrol was found to act as an antioxidant and antimutagen and to induce phase II drug-metabolizing enzymes (anti-initiation activity); it mediated anti-inflammatory effects and inhibited cyclooxygenase and hydroperoxidase functions (antipromotion activity); and it induced human promyelocytic leukemia cell differentiation (antiprogression activity). In addition, it inhibited the development of preneoplastic lesions in carcinogen-treated mouse mammary glands in culture and inhibited tumorigenesis in a mouse skin cancer model. These data suggest that resveratrol, a common constituent of the human diet, merits investigation as a potential cancer chemopreventive agent in humans.

Animals↗

Comparative linkage mapping of human chromosome 13 and bovine chromosome 12.

A comparative linkage map of human chromosome 13 and bovine chromosome 12 was constructed using eight polymorphic microsatellite markers associated with six specific genes. Linkage of these was also examined relative to five previously mapped anonymous microsatellite markers. Seven gene-linked markers were developed from bovine large-insert genomic clones containing one of five genes of interest (serotonin receptor subtype 2, fms-related tyrosine kinase, coagulation factor 10, retinoblastoma susceptibility gene, collagen type IV alpha 1), and one additional marker was developed from a microsatellite resident within an intron of the bovine dopachrome tautomerase gene. Four of these loci were previously assigned to bovine chromosome 12 by analysis of a somatic cell hybrid panel. This study provides linkage information for examining gene order in this conserved synteny group. The comparative linkage mapping results indicate that the q arm of human chromosome 13 is almost entirely conserved in bovine chromosome 12. One intrachromosomal rearrangement was detected in this linkage group relative to human, and this rearrangement was confirmed by fluorescence in situ hybridization results.

Animals↗

Energetics of the lattice: packing elements in crystals of four-stranded intercalated cytosine-rich DNA molecules.

Condensation of single molecules from solution into crystals represents a transition between distinct energetic states. In solution, the atomic interactions within the molecule dominate. In the crystalline state, however, a set of additional interactions are formed between molecules in close contact in the lattice--these are the packing interactions. The crystal structures of d(CCCT), d(TAACCC), d(CCCAAT), and d(AACCCC) have in common a four-stranded intercalated cytosine segment, built by stacked layers of cytosine.cytosine+ (C.C+) base pairs coming from two parallel duplexes that intercalate into each other with opposite polarity. The intercalated cytosine segments in these structures are similar in their geometry, even though the sequences crystallized in different space groups. In the crystals, adenine and thymine residues of the sequences are used to build the three-dimensional crystal lattice by elaborately interacting with symmetry-related molecules. The packing elements observed provide novel insight about the copious ways in which nucleic acid molecules can interact with each other--for example, when folded in more complicated higher order structures, such as mRNA and chromatin.

Base Sequence↗

Bipolar esophageal electrocardiography by using standard limb leads.

The ear-xiphisternum distance (EXD, the distance from the low edge of the ear to the xiphisternal basis in supine position) was used as a reference value for esophageal catheter insertion. ECGs recorded in the esophagus with bipolar electrocardiography using standard limb lead (ESLL) and conventional unipolar lead (ECUL) were compared. 112 patients with sinus rhythm and 76 patients during paroxysmal supraventricular tachycardia (PSVT) whose P-wave and QRS complex did not overlap were studied. The results suggested that in sinus rhythm the amplitude of the P-wave in ESLL was larger and the T-wave was smaller than in ECUL. During PSVT, the P-wave was much clear and higher in each lead of ESLL than that in ECUL. The ideal range of esophageal ECG recording was situated between the end of EXD and 6.5 cm proximal to it.

Adolescent↗

Antimicrobial compounds from Ceanothus americanus against oral pathogens.

During the search for antimicrobial compounds from higher plant sources, a methanol extract of Ceanothus americanus demonstrated antimicrobial activity against selected oral pathogens. Through further bioassay-guided fractionation and purification, three triterpenes (ceanothic acid, 27-hydroxy ceanothic acid and ceanothetric acid) and two flavonoids (maesopsin and maesopsin-6-O-glucoside) were identified. Among these, ceanothetric acid and maesopsin-6-O-glucoside were new compounds. Ceanothic acid and ceanothetric acid demonstrated growth inhibitory effect against Streptococcus mutans, Actinomyces viscosus, Porphyromonas gingivalis, and Prevotella intermedia with MICs ranging from 42 to 625 micrograms ml-1. Maesopsin, its glucoside, and 27-hydroxy ceanothic acid, were inactive below the concentration of 500 micrograms ml-1.

Actinomyces viscosus↗

Induction of apoptosis in the germ cells of adult male rats after exposure to cyclophosphamide.

Treatment with cyclophosphamide, a commonly used anticancer drug, may result in oligozoospermia or azoospermia. The objective of this study was to determine whether exposure of male rats to cyclophosphamide induces apoptosis in male germ cells, and if so, when the peak of apoptosis occurs and at what specific stages of spermatogenesis. The presence of apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) detection in situ and by an increase in DNA fragmentation (DNA ladder). To determine the time course of drug-induced apoptosis, male Sprague-Dawley rats were treated with a single dose (70 mg/kg BW) of cyclophosphamide, and the testes were fixed 0, 4, 8, 12, 18, 24, and 48 h after treatment. To determine the dose response, rats were treated with doses of cyclophosphamide (0, 2, 7, 20, and 70 mg/kg), and the testes were fixed 12 h after treatment. A low spontaneous incidence of apoptosis was observed in controls, in particular in premeiotic germ cells of stages I-IV and XI-XIV of the seminiferous tubules. In cyclophosphamide-exposed rats, the incidence of apoptosis increased progressively at 4 h and 8 h, reached a peak at 12 h (about 3.5-fold above control), and then decreased rapidly to control levels by 48 h. A 70-mg/kg dose of cyclophosphamide induced a significant increase in apoptosis; lower doses did not. Although drug-induced apoptosis occurred in all stages of germ cells, it was most pronounced in spermatogonia and spermatocytes in stages I-IV and XI-XIV. Thus, apoptosis may be involved in the occurrence of oligozoospermia or azoospermia after cyclophosphamide treatment. Apoptosis of damaged premeiotic germ cells may serve a critical role in protecting subsequent generations from the diverse effects of toxicants.

Animals↗

[The cloning, expression, purification and bioactivity identification of recombinant IGF-I].

The fragment containing IGF-I cDNA and cyc1 terminator from PSK-IGF-I plasmid was cloned into PSK43 SB treated by Hind III, Klenow and Cla I, generating a recombinant PSB-IGF-I (DH), which have double Hind III sites. This plasmid was treated by Hind III and Klenow, ligated to give a recombinant plasmid PSB-IGF-I(NH), which has no Hind III site. After analysed by restriction endonucleases and sequencing, the fusion site was verified to be correct. This plasmid was excised and a BamH I-Cla I fragment containing yeast alpha-factor promotor, leader sequence, IGF-I cDNA and cycl terminator was cloned into yeast episomal plasmid vector YEpHC8, generating a recombinant plasmid YEpHC8-IGF-I. Transformed this plasmid into yeast competence cells BJ1990 by using LiAc method. The expressed products were secrected into the medium broth and having the correct molecular weight of 8,100 on SDS-PAGE. The products of IGF-I were raised by using high-cell-density fermentation of Saccharomyces cerevisiae. The cleared supernatant of yeast medium was applied to Bio-Rex 70 resin, eluted with 1 mol/L ammonium acetate, pH8.0. Main peak was pooled and applied to Bio-Gel P10 (200-400 mesh). The second peak was collected and get relatively pure IGF-I proteins. The biological activities of IGF I product was assayed in NIH3T3 cells by using MTT method. The results show that the expressed IGF-I can obviously stimulate NIH3T3 cells to proliferate at the concentration ranging from 10 ng/ml to 50 ng/ml, suggesting that the protein has its biological activities.

3T3 Cells↗