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Using a BLAST-searching approach, we identified a mouse expressed sequence tag (EST) clone (AA038672) showing great similarity to the 3' end of the human MUC5B gene. The clone was named "3pmmuc5b-1" after complete nucleotide sequencing (Genbank Accession, AF369933). A subsequent search of the mouse genome database with the 3pmmuc5b-1 sequence identified two overlapping genomic clones (AC020817 and AC020794) that contained the sequence of both 3pmmuc5b-1 and the mouse Muc5ac gene. Like their human homologs, the genomic order of the mouse Muc genes is 5'-Muc5ac-Muc5b-3'. These results suggest that the newly identified EST clone, 3pmmuc5b-1, is part of the 3' portion of the mouse Muc5b gene. In situ hybridization demonstrated that this putative mouse Muc5b message was expressed in a restricted manner in the sublingual gland region of the tongue and the submucosal gland region of the mouse trachea in a normal animal. However, the gene expression was greatly enhanced in airway surface epithelium and the submucosal gland region in ovalbumin-induced asthmatic mice. These results were consistent with previous studies of human airway tissues. We therefore conclude that this newly cloned mouse Muc5b gene could be used as a marker for studying aberrant mucin gene expression in mouse models of various airway diseases.

We report linear free energy relation (LFER) models of the equilibrium distribution of molecules between blood and brain, as log BB values. This method relates log BB values to fundamental molecular properties, such as hydrogen bonding capability, polarity/polarisability and size. Our best model of this form covers 148 compounds, the largest set of log BB data yet used in such a model, resulting in R(2)=0.745 and e.s.d.=0.343 after inclusion of an indicator variable for carboxylic acids. This represents rather better accuracy than a number of previously reported models based on subsets of our data. The model also reveals the factors that affect log BB: molecular size and dispersion effects increase brain uptake, while polarity/polarisability and hydrogen-bond acidity and basicity decrease it. By splitting the full data set into several randomly selected training and test sets, we conclude that such a model can predict log BB values with an accuracy of less than 0.35 log units. The method is very rapid-log BB can be calculated from structure at a rate of 700 molecules per minute on a silicon graphics O(2).

Objective: To investigate ultrastructural pathological changes of Heroin-Addicts., Methods: Heroin-Addicts' central nervous system, endocrine system, immune system and reproductive system in 4 cases are observed by using transmission electron microscope(TEM)., Results: The changes of central nervous system are mitochondrion swelling, crista fragmentation and disappear. Endoplasmic reticulum dilation, nervous fibres and cell organelles reduction; mitochondrion swelling, Partial crista fragmentation and endoplasmic reticulum dilation are also found in endocrine system; Lymphocytes reduction, cytoplasm ingredient reduction and dead lymphocytes increase in immune system; in reproductive system, spermatogenic cells and cell organelles are reduced in the male and follicle disappeared in the female., Conclusion: Ultra-structural pathological changes of heroin-addicts are presented acute, chronic oxygen deficiency degeneration and necrosis.

50% effective inhibition concentration 48h-EC50 of 40 substituted benzenes to the algae (Scenedesmus obliquus) was determined. The energy of the lowest unoccupied molecular orbital (E(LUMO)) was calculated by the quantum chemical method MOPAC6.0-AM1. By using E(LUMO) and the hydrophobicity parameter log K(OW) the quantitative structure-activity relationship model (QSAR) was developed: log1/EC50=0.272 logK(OW) - 0.659E(LUMO) + 2.54, R2 = 0.793, S.E. = 0.316, F = 71.07, n = 40. A series of equations were obtained about the measured EC50 values of different subclasses of compounds. For those compounds containing double -NO2, their toxicity may be related chiefly to the intracellular reduction of -NO2 obtaining electron, while for anilines and phenols, K(OW) contributes most to the QSAR and E(LUMO) very little.

Multiple retinoic acid responsive cDNAs were isolated from a high density cDNA microarray membrane, which was developed from a cDNA library of human tracheobronchial epithelial cells. Five selected cDNA clones encoded the sequence of the same novel gene. The predicted open reading frame of the novel gene encoded a protein of 319 amino acids. The deduced amino acid sequence contains four motifs that are conserved in the short-chain alcohol dehydrogenase/reductase (SDR) family of proteins. The novel gene shows the greatest homology to a group of dehydrogenases that can oxidize retinol (retinol dehydrogenases). The mRNA of the novel gene was found in trachea, colon, tongue, and esophagus. In situ hybridization of airway tissue sections demonstrated epithelial cell-specific gene expression, especially in the ciliated cell type. Both all-trans-retinoic acid and 9-cis-retinoic acid were able to elevate the expression of the novel gene in primary human tracheobronchial epithelial cells in vitro. This elevation coincided with an enhanced retinol metabolism in these cultures. COS cells transfected with an expression construct of the novel gene were also elevated in the metabolism of retinol. The results suggested that the novel gene represents a new member of the SDR family that may play a critical role in retinol metabolism in airway epithelia as well as in other epithelia of colon, tongue, and esophagus.

The human intestinal absorption of 241 drugs was evaluated. Three main methods were used to determine the human intestinal absorption: bioavailability, percentage of urinary excretion of drug-related material following oral administration, and the ratio of cumulative urinary excretion of drug-related material following oral and intravenous administration. The general solvation equation developed by Abraham's group was used to model the human intestinal absorption data of 169 drugs we considered to have reliable data. The model contains five Abraham descriptors calculated by the ABSOLV program. The results show that Abraham descriptors can successfully predict human intestinal absorption if the human absorption data is carefully classified based on solubility and administration dose to humans., (Copyright 2001 Wiley-Liss, Inc.)

The study was undertaken to find whether microinjection of vasopressin (AVP) into the rostral ventrolateral medulla (rVLM) affects blood pressure (BP) and blood viscosity in rats and to analyze the effects of AVP-ergic mechanism in the rVLM on stress-induced hyperviscosity of rats. The results are as follows. (1) Microinjection of AVP into the rostral ventrolateral medulla (rVLM) of normal rats resulted in a significant increase in BP and plasma viscosity, which was blocked by bilateral microinjection of AVP V(1) receptor antagonist (d(CH2)5 Tyr(Me)(2) AVP) into the same area. (2) The stress-induced pressor response and hyperviscosity was abolished by microinjection of AVP V(1) receptor antagonist into the rVLM, but not by AVP V(2) receptor antagonist ( adamantaneacetyl(1),O-Et-D-Tyr(2),Val(4),aminobutyryl(6),Arg(8,9) -vasopressin). Microinjection of AVP V(1) receptor antagonist of the same dosage into bilateral caudal ventrolateral medulla (cVLM) or intravenous injection of the antagonist of the same dosage had no effect. The above results suggest that the increase of BP and plasma viscosity induced by microinjection of AVP into rVLM and the stress-induced pressor response and hyperviscosity caused by restraining may be mediated by activation of the AVP V(1) receptor in the rVLM.

The presence of seven stress proteins including various heat shock proteins [27-kDa (HSP27), 60-kDa (HSP60), 70-kDa (HSP70) and its constitutive form HSC70, and 90-kDa (HSP90) HSPs] and two glucose-regulated proteins [75-kDa (GRP75) and 78-kDa (GRP78) GRPs] in ozone-exposed lungs of nonhuman primates and in cultured tracheobronchial epithelial cells was examined immunohistochemically by various monoclonal antibodies. Heat treatment (42 degrees C) resulted in increased HSP70, HSP60, and HSP27 and slightly increased HSC70 and GRP75 but no increase in GRP78 in primary cultures of monkey tracheobronchial epithelial cells. Ozone exposure did not elevate the expression of these HSPs and GRPs. All of these HSPs including HSP90, which was undetectable in vitro, were suppressed in vivo in monkey respiratory epithelial cells after ozone exposure. Both GRP75 and GRP78 were very low in control cells, and ozone exposure in vivo significantly elevated these proteins. These results suggest that the stress mechanism exerted on pulmonary epithelial cells by ozone is quite different from that induced by heat. Furthermore, differences between in vitro and in vivo with regard to activation of HSPs and GRPs suggest a secondary mechanism in vivo, perhaps related to inflammatory response after ozone exposure.

The toxicities of benzoic acids to Vibrio fischeri, Daphnia magna and carp were measured. The results showed that the toxicity to V. fischeri and Daphnia decreased in the order of bromo > chloro > fluoro approximately equal to aminobenzoic acids. The toxicity of substituted benzoic acids to carp and Daphnia was much lower that to V. fischeri. The results also showed that the toxicity of benzoic acids to Daphnia decreased as the pH increased. It is suggested that ionized and non-ionized forms have different toxic responses. The non-ionized form may play an important role in toxicity because the toxicity of benzoic acids to Daphnia greatly decreases as the pH increases. The toxicity of benzoic acids to Daphnia may operate through non-polar narcosis, based on the regression results between the toxicities and partition coefficients (log P) and apparent partition coefficients (log D). However, toxicity cannot be predicted from non-polar baseline models because the ionized and non-ionized form of benzoic acids have different contributions to toxicity. Compared with the single descriptors, the prediction of toxicity of the benzoic acids was improved remarkably by using log P with pKa and log P with ELUMO. For the toxicity of benzoic acids to V. fischeri, it is suggested that the toxic mechanism may be different from the mechanism in Daphnia and carp. A probable reason is that V. fischeri is a unicellular organism with low lipid content, and hence both ionized and non-ionized forms of benzoic acids can easily cross the cell membrane and contribute to toxicity.

The development of routine techniques for the isolation and in vitro maintenance of conducting airway epithelial cells in a differentiated state provides an ideal model to study the factors involved in the regulation of the expression of mucociliary differentiation. Several key factors and conditions have been identified. These factors and conditions include the use of biphasic culture technique to achieve mucociliary differentiation, the use of such stimulators as the thickness of collagen gel substratum, the calcium level, and vitamin A, and such inhibitors as the growth factors, epidermal growth factor and insulin, and steroid hormones, for mucous cell differentiation. Using the defined culture medium, the life cycle of the mucous cell population in vitro has been investigated. It was demonstrated that the majority of the mucous cell population in primary cultures is not involved in the replication of deoxyribonucleic acid (DNA). However, the mucous cell type is capable of self-renewal in culture, and this reproduction is vitamin A dependent. Furthermore, differentiation from nonmucous to mucous cell type can be demonstrated by adding back a positive regulator such as vitamin A to the "starved" culture. Cell kinetics data suggest that vitamin A-dependent mucous cell differentiation in culture is a DNA replication-independent process, and the process is inhibited by transforming growth factor-beta1.

Aim: To investigate the morphology of lymph vessel capillaries in both human gastric carcinomas and their peritumoral tissues, as well as the relation of this morphology to lymphatic metastasis., Methods: The morphology and fine distribution of both lymph vessels and capillaries in and around the primary foci of gastric carcinoma were studied using the 5'Nase Alpase double staining method. The total amount of opened vessels and the opening rate of lymph vessels and capillaries were counted using a light microscope (100 × magnification), and the maximal luminal area, perimeter and diameter were measured using an image analysis technique., Results: Lymph vessels and capillaries displayed strong 5'Nase-positive staining (brown and dark brown), while blood vessels and capillaries revealed significant Alpase activity (blue). There were many lymph vessels, capillaries and solid strip-like tissues found in the gastric carcinoma samples analyzed. The total amount of lymphatics in the metastatic group (gastric carcinoma vs peritumoral tissue) and non-metastatic group was 26.9 ± 14.2 vs 10.4 ± 4.0, 11.4 ± 3.4 and 9.7 ± 3.2, P < 0.01, respectively. Their opening rates were 21.2 vs 47.5 and 40.4 vs 46.0, P < 0.01, respectively. Their maximal luminal areas were 1502.98 ± 1236.91 vs 5526.80 ± 4853.42; 1918.14 ± 2299.24 vs 3836.16 ± 3549.16; 5526.80 ± 4853.42 vs 3836.16 ± 3549.16, P < 0.05, their perimeters were 220.33 ± 130.25 vs 441.43 ± 276.51; 241.79 ± 171.13 vs 333.80 ± 199.66; 441.43 ± 276.51 vs 333.80 ± 199.60, P < 0.05, and their diameters were 28.80 ± 14.98 vs 59.39 ± 28.53; 25.37 ± 15.79 vs 46.22 ± 20.85; 59.39 ± 28.53 vs 46.22 ± 20.85, P < 0.05, respectively. In summary, the lymphatics found in gastric carcinoma samples from the metastatic group were significantly lower than those of the other groups., Conclusion: There are newly formed lymph capillaries found in gastric carcinoma. Dilation of lymph capillaries may be related to edema found in peritumoral connective tissues. The observed lymph node metastases from gastric carcinoma occur through mature lymph capillaries that invade in and around primary gastric carcinoma foci.

A palindromic primer-based mRNA differential display method has been used to isolate various vitamin A-responsive genes from primary cultures of monkey tracheobronchial epithelial cells. This method, as compared with the original mRNA differential display (mDD) method described by Liang and Pardee, used only one arbitrarily designed primer instead of two in the polymerase chain reaction. The single-primer mDD method has several advantages over the two-primer mDD system, especially in the reamplification and the selection of 5'-end cDNA clone. To verify the usefulness of this approach, one of these differential display bands, M34, was initially chosen for further amplification and cloning. The clone derived from the M34 band has a DNA sequence with > 90% homology to the human nucleolin gene. Furthermore, DNA sequencing confirms that both 5' and 3' ends of the insert of M34 contain the invertly repetitive nucleotide sequence that was used to direct this cloning. Nucleolin is a multifunctional phosphoprotein that plays an important role in ribosome biogenesis and mRNA stability. Northern blot analysis demonstrated that in addition to the elevation by vitamin A, the level of nucleolin message is significantly higher in fetal than in adult tracheobronchial epithelial cultures. Furthermore, in situ hybridization demonstrated that the amount of nucleolin message is significantly higher in both basal and ciliated cell types than in mucous and intermediary cell types. These results support the feasibility that the single-primer mDD technique can be used to isolate vitamin A-responsive genes with a palindromic nature.

The autocrine/paracrine growth mechanism has been implicated in the regulation of bronchial epithelial cell proliferation. By inhibiting the expression of the transforming growth factor-alpha (TGF-alpha) gene product, vitamin A is able to suppress the proliferation of tracheobronchial epithelial cells in culture. Similar repressions in TGF-alpha mRNA levels by retinol were observed in airway explant cultures and in a cell line immortalized from normal human bronchial epithelial cells. Both the nuclear run-on transcriptional assay and the transfection study with the chimeric construct of the TGF-alpha promoter and chloramphenicol acetyltransferase reporter gene partly suggest a transcriptional downregulation mechanism of TGF-alpha gene expression by the retinol treatment; however, this inhibition at the transcriptional level cannot account for the total inhibition at the mRNA level. These results suggest that a downregulation of the expression of the TGF-alpha gene at the transcriptional and post-transcriptional levels by vitamin A may precede the essential event associated with the homeostasis of normal conducting airway epithelium.

Restriction fragment length polymorphism (RFLP) markers were used to map male fertility restoring gene that was transferred from chromosome 6U of Aegilops umbellulata Zhuk. to wheat. Segments of chromosome 6U bearing the gene that restore fertility to T. timopheevi Zhuk. male sterile cytoplasm were identified in all four translocation lines by two probes, BCD21 and BCD342. Lines 040-5, 061-1 and 061-4 are T6BL.6BS-6U translocations, while line 2114 is a T6AL.6AS-6U translocation. Line 2114 has a much larger 6U chromosomal segment and lower frequency of transmission of male gametes with the alien segment than the other three lines. The restoring gene carried by the 6U segment in 2114 showed high expressivity and complete penetrance. This restoring gene is designated Rf6. A homoeologous chromosome recombination mechanism is discussed for the alien gene transfer.

38 cases of acute leukemia were treated with autologous bone marrow transplantation (ABMT). The marrow was purged with hyperthermia of 42 degrees C in vitro for one hour in microwave. Twenty-two among them were AML (CR1). Sixteen were ALL (CR1). The mean age was 26 (10-43) years. All the patients were engrafted successfully after ABMT. Mean DFS was 21 (3-69) months. Four cases relapsed at 3 to 8 months after ABMT. Two patients with ALL developed central nervous system leukemia at 12 and 15 months respectively after ABMT. The DFS and probability of relapse at 5 years were 67.8% and 16.8% respectively for all patients.

The cellular localization of the tyrosine-specific protein kinase p62c-yes, the product of the proto-oncogene c-yes, has been examined in the striatonigral neurons which interconnect the rat neostriatum and substantia nigra. Although p62c-yes was more enriched in the neostriatum than in the substantia nigra, excitotoxin-induced necrosis of nerve cells in the neostriatum led to 50-60% decreases of p62c-yes both in the lesioned neostriatum and in the ipsilateral substantia nigra. Hence, the p62c-yes tyrosine kinase is present both in the cell body region and in the axonal and nerve terminal region of the striatonigral neurons. This localization indicates that the enzyme may be involved in both presynaptic and postsynaptic functions in mammalian forebrain neurons.

In this paper, we present a conductivity method for measuring the RBC (red blood cell) aggregation-sedimentation phenomenon and the theoretical analysis of this phenomenon. A theory describing the RBC aggregation-sedimentation phenomenon with the conductivity method is proposed and the formula of the l-t function which depicts the RBC aggregation-sedimentation process is given. We put forward the aggregation index and sedimentation factor describing the RBC aggregation-sedimentation phenomenon for the first time.

To study the causes of osteopenia after plate fixation in the treatment of fractures and bone diseases, intact tibiae of 26 dogs were fixed by 3 different plates at various rigidities. The dogs were killed at 24 hours 3, 6, 12, 20 weeks. Disulphine blue was injected intravenously before killing. The results showed that in addition to stress shielding the disturbance of blood supply was the main cause of osteopenia in the early stage. Reaction to the implant might be another cause in the middle stage. The disuse of the affected limb was the most important factor in the late stage. Osteopenia can be treated by active exercises and normal weight-bearing before plate removal.

Expression of the c-yes proto-oncogene in mammalian brain, retina and adrenal gland was studied by immunohistochemistry and immune assays with affinity-purified anti-yes IgG. Immunohistochemical staining with anti-yes IgG showed that in the central nervous system p62c-yes is highly expressed in mitral cells of the olfactory bulb, Purkinje cells of the cerebellum, hippocampal neurons and granule neurons of the dentate gyrus and ependymal cells lining central nervous system ventricles. Less intense labeling by anti-yes IgG was observed in most neuronal cell bodies in the brain. In addition, we observed intense c-yes immunoreactivity in the ganglion cells of the retina, the inner segment layer of rods and cones and medullary cells of the adrenal gland. Mapping p62c-yes expression to specific areas of mammalian neural tissues points to attractive experimental systems which could be used to investigate the function of the proto-oncogene product in neural processes.

The protein product of the c-yes proto-oncogene, p62c-yes, is highly expressed in a variety of mammalian cell types, including neurons, spermatozoa, platelets, and epithelial cells. In order to understand the function of p62c-yes in epithelial cells, the expression and localization of p62c-yes was studied in cultured human epidermal keratinocytes and in normal, hyperplastic, and neoplastic human epidermis. Human keratinocytes in culture produce a single 4kb c-yes mRNA and a 62kd protein product, p62c-yes, which is active as a protein tyrosine kinase. Using affinity-purified antibodies generated to the amino-terminus of the human c-yes protein, the expression of p62c-yes was localized to keratinocytes in the basal epidermal layer of normal neonatal and adult epidermis. There was a marked reduction in expression of p62c-yes by suprabasal keratinocytes undergoing progressive differentiation. By immunofluorescence microscopy, p62c-yes was localized to the plasma membrane and to a perinuclear cytoplasmic area in cultured keratinocytes. The apparent association of p62c-yes with plasma membranes was particularly evident in suprabasal keratinocytes from hyperplastic epidermis. Neoplastic keratinocytes in basal cell carcinomas showed a marked reduction in p62c-yes expression compared to normal basal keratinocytes in epidermis or to proliferating cultured keratinocytes. Thus the expression of p62c-yes by one epithelial cell type, the keratinocyte, is altered by cellular differentiation and neoplastic transformation. Keratinocytes provide a normal epithelial cell model in which the biochemical function of p62c-yes can be studied.

We are prospectively examining the relation between environmental lead exposure and pregnancy outcome in cohorts of women exposed to a wide range of air lead concentrations. Titova Mitrovica, Yugoslavia, is the site of a large lead smelter, refinery, and battery factory. At midpregnancy, 602 women in T. Mitrovica and 900 women in Pristina, a non-lead-exposed control town, were interviewed. Blood was obtained for blood lead (PbB), hemoglobin, erythrocyte protoporphyrin, and serum ferritin measurements. Women were seen again at delivery, at which time maternal and umbilical cord blood samples were obtained. While many demographic and social characteristics were similar across the two towns, women in Pristina were more likely to report employment outside the home, cigarette smoking, and alcohol use during pregnancy. As expected, PbB levels were substantially higher in the smelter town. At midpregnancy, PbB geometric means were 17.1 micrograms/dL in T. Mitrovica and 5.1 micrograms/dL in Pristina; 86% of the pregnant women in T. Mitrovica, compared to 3.4% of those in Pristina, had PbB levels greater than 10 micrograms/dL. Within T. Mitrovica, distance between the home and the smelter was the most important predictor of PbB at mid-pregnancy and delivery. Husband's employment in the lead industry was associated with a significant increase in maternal PbB levels independent of place of residence. Higher maternal serum ferritin concentrations were associated with lower PbB levels, suggesting that dietary iron inhibits lead absorption. Overall, the placenta was a poor barrier to lead; the relationship between maternal PbB and umbilical cord PbB was linear across a wide range of PbB levels.

Expression of the c-yes proto-oncogene in normal mammalian tissues was studied by immunohistochemistry and by immunoprecipitation assays using affinity purified anti-yes IgG. The antibody was raised against the amino-terminal domain of the human c-yes protein expressed in bacteria as a trpE-yes fusion protein. We detected p62c-yes in a variety of rat and human tissues but its relative level of expression varied significantly among different cell types. Certain epithelial cells, platelets and spermatid acrosomes showed the highest levels of p62c-yes. Intracellular localization of p62c-yes in epithelial cells differed among organs performing different physiological functions. The overall pattern of expression suggests that p62c-yes may be involved in numerous cellular pathways.

Trigeminal neuralgia is often seen in elderly persons. Two hundred patients with trigeminal neuralgia were submitted to microvascular decompression by a retrosigmoid approach. The surgical procedure is described. Vascular compression of the nerves was noticed in all these patients at operation. One hundred ninety-seven had relief of symptoms postoperatively, for a 98.5% cure rate. In addition, we suggest that another important cause is arteriosclerosis.

Systemic investigations of chromium determination in urine with diphenylcarbazide were carried out and suitable analytic conditions for the determination established. Urine samples were first digested with nitric acid-hydrogen peroxide and then the chromium in the sample was oxidized to the hexavalent state by permanganate in acidic medium. After the excess oxidizing reagent was destroyed with sodium nitrite, chromium (VI) combined with diphenylcarbazide, forming a stable red color. The average recovery was 101.1%, ranging from 82.0% to 115.8%. Coefficients of variation were in the range from 2.0% to 8.4%. The method can be conveniently applied to the analysis of trace chromium in urine samples with satisfactory sensitivity and recovery. The method was applied to the measurement of 180 normal and 124 CHD (coronary heart disease) urine samples from Chengdu city. The mean of chromium level in normal urine was 6.1 micrograms/L and the range with 95% confidence of urine chromium was determined as 1.7-11 micrograms/L. The mean of chromium level in CHD urine samples was 4.2 micrograms/L. By statistical analysis, there was a significant difference between the means of the two groups (t' = 6.9, P less than 0.05).

The cytology of nine new D genome hybrids involving Triticum syriacum, Triticum ventricosum, Triticum cyclindricum, Triticum juvenale, Triticum crassum, Triticum tauschii and Triticum aestivum is described. The calculation of numerical values of the relative affinity and the patterns of chromosome pairing indicate that the D genome in T. syriacum and T. juvenale may have been substantially modified and that of T. crassum somewhat modified from that of the diploid progenitor, T. tauschii.

The element contents in the soil where the Jilin Big-Wild Ginseng grows were determined with plasma emission spectrometry. 23 elements in the soil were found, of which 12 are macro and trace elements indispensable to Ginseng growth. This study provides a scientific basis for selecting suitable soil for planting wild ginseng, or for creating optimum soil conditions for the bumper harvest and high quality of Ginseng.