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Biomedical subjects

Q Fan

Publications and source records attributed to Q Fan.

At least 37 records · Page 2Linked to original sources

Cellular terrain surrounding sympathetic nerve pathways in the rat orbit: comparisons of orbital connective tissue and smooth muscle cell phenotypes.

Sympathetic axons are abundant within some orbital tissues but are absent from others. This study investigated cellular phenotypes of tissues containing sympathetic nerves en passage and compared these with phenotypes in regions devoid of sympathetic nerves and with smooth muscle targets. Two primary orbital smooth muscle targets, the tarsal muscle and orbital muscle, contained many synaptophysin-immunoreactive nerves. Target cells had ultrastructural features typical of smooth muscle and were immunoreactive for alpha-smooth muscle actin, smooth muscle myosin heavy chain, desmin, vinculin, and laminin, but not non-muscle myosin, vimentin, fibronectin, or type IV collagen; nerve growth factor (NGF) mRNA was detected by reverse transcription-polymerase chain reaction. Periorbital sheath devoid of sympathetic nerves contained elongated fibroblasts that were immunoreactive for vimentin, non-muscle myosin, and fibronectin, but not for alpha-smooth muscle actin, smooth muscle myosin heavy chain, vinculin, desmin, laminin, or type IV collagen, and did not express NGF mRNA. Regions of periorbital sheath containing sympathetic nerves had few synaptophysin-immunoreactive varicosities. Cells in this region contained myofilaments, ribosomes, and rough endoplasmic reticulum and were larger than tarsal muscle cells. They expressed NGF mRNA and showed a unique immunophenotype, reacting for vimentin, alpha-smooth muscle actin and myosin heavy chain, desmin, vinculin, laminin, and type IV collagen. This phenotype reflects both fibroblast and smooth muscle features similar to myofibroblasts or transdifferentiated smooth muscle described in other tissues. The spatial association between these cells and sympathetic nerves suggests that they may be involved in axon guidance or maintenance.

Animals↗

Variations in uterine innervation during the estrous cycle in rat.

Uterine innervation undergoes profound remodeling during puberty, pregnancy, and after parturition. However, the extent to which uterine innervation may change during the estrous cycle is uncertain. The objective of the present study was to determine whether nerve fiber density of the uterine horn is altered during the estrous cycle and, if so, which subpopulations are affected. Immunostaining for the pan-neuronal marker protein gene product (PGP) 9.5 revealed fibers within the vascular zone, myometrium, and endometrium, with greater density in the ovarian and cervical regions than in the middle. In most structures, nerve density was reduced during estrus. This could not be accounted for by increased target volume, as the reduction in longitudinal muscle innervation persisted after correction for changes in target size. Immunostaining for vasoactive intestinal polypeptide-immunoreactive parasympathetic nerves revealed fibers associated predominantly with the vascular zone and circular muscle within the cervical region. No cyclical variation was detected. Calcitonin gene-related peptide-immunoreactive nerves were present within all structures, and density was highest at the ovarian end. These fibers also did not vary significantly through estrous. Dopamine beta-hydroxylase-immunoreactive sympathetic nerves innervated all structures, with greater density in the ovarian end. These fibers were reduced substantially during estrus, but the decline was also significant in proestrus, thus preceding that detected by using the pan-neuronal marker. We conclude that the estrous cycle in rat is accompanied by structural remodeling of sympathetic nerves by way of retraction or degeneration of terminal fibers during estrus. The structural loss of the terminal axon apparently is preceded by depletion of catecholamine-synthesizing enzyme.

Animals↗

Antisense in abundance: the ribosome as a vehicle for antisense RNA.

Insertions at some sites within rRNA variable regions can be tolerated without affecting rRNA function. Antisense RNAs inserted at such sites in the T. thermophila rRNA can eliminate phenotypically or immunologically detectable gene expression of three genes tested. This unusually effective antisense activity is probably due to the abundance, stability and favourable intracellular localization of these antisense rRNAs with respect to mRNAs. Since antisense RNAs function very well as a part of the rRNA, rRNA might also be useful as a vehicle for other RNAs that might affect cell function such as protein binding sites or trans-acting ribozymes. The robust function of the antisense ribosome system in T. thermophila should allow the use of this system to specifically suppress gene expression and to clone genes by their null or hypomorphic phenotypes. The use of the antisense ribosome in other eukaryotes has yet to be explored, but the realization of this goal is well within the realm of possibility.

Animals↗

Augmentation of recombinant fibronectin polypeptide CH50 on the antitumor function of macrophages.

We prepared an anti-metastatic polypeptide, recombinant fibronectin polypeptide CH50, and finished the preliminary identification of its functions. In this paper, we studied the effect of this polypeptide on the function of macrophages. CH50 can significantly augment the production of nitric oxide(NO) by macrophages in a dose-dependent manner. The continuous presence of CH50 had a much stronger effect. In the presence of CH50, the cytotoxicity of macrophages to melanoma B16/F1 cells was significantly enhanced, and a stronger effect was obtained if CH50 was present continuously. CH50 polypeptide and IFN-gamma have a synergistic effect on the production of NO by macrophages and the cytotoxicity of macrophages on tumor cells. In the in vivo experiments, CH50 can inhibit the growth of tumor cells, and have a better effect in the presence of IFN-gamma. Our results suggest that recombinant fibronectin polypeptide CH50 has two functions: one is to inhibit the metastasis of tumor cells, and the other one is to augment the function of macrophages. And this polypeptide will be potentially useful in tumor therapy.

Animals↗

Effect of fragment Asp1961-Glu1978 in fibronectin on the expression of triple-domain polypeptide in E. coli.

Two plasmids were constructed and used to express two triple-domain recombinant polypeptide of human fibronectin (FN). The cDNAs in plasmids code for two polypeptides, CH62 (Pro1239-Ser1515 of FN linked with Ala1690-Val2049 through Met) and CH63 (CH62 without Ile1850-Glu1978). The expression level of CH62 in E. coli was very low, but that of CH63 was very high. The results suggests that Asp1961-Glu1978 in FN is a key sequence influencing the expression of triple-domain polypeptide in E. Coli. After being dissolved and renatured, CH63 can be purified by heparin-agarose affinity chromatography. Both of the cell-binding domains in the recombinant polypeptide were functional. The production of CH63 provides a fundamental basis for further study of recombinant products with better anti-metastasis function.

Animals↗

Photodynamic therapy of B16F10 murine melanoma with lutetium texaphyrin.

Photodynamic therapy (PDT) of pigmented melanoma has generally been unsuccessful because of insufficient light penetration in such tissues. In this study, the responsiveness of the heavily pigmented B16F10 murine melanoma to lutetium texaphyrin (PCI-0123), a water-soluble sensitizer with strong absorbance in the near infrared (700-760 nm), was examined. These studies were carried out in both normal and ApoE deficient C57BL/6 mice. The latter strain exhibits a lipoprotein profile more like humans (low density lipoprotein > high density lipoprotein) than rodents (high density lipoprotein >> low density lipoprotein). Under optimal conditions of drug dose, light dose, and interval between drug administration and irradiation--the median survival time of C57BL/6 tumor bearing mice was approximately doubled (29 d) compared with tumor bearing control animals (13 d). The life-span of the ApoE knockout mice was greater (33 d) than the C57BL/6 animals (23 d) when irradiation occurred 3 h after administration of a 10 micromol per kg drug dose. The greater efficacy of PDT in the ApoE deficient mice was associated with more rapid clearance of drug from the blood, greater accumulation of sensitizer in tumor tissue, and substantially greater drug binding to the very low density lipoprotein/low density lipoprotein plasma fraction. Confocal laser scanning microscopy showed that the predominant subcellular site of photosensitizer binding was to melanosomes; costaining was performed with Mel-5. Melanosomes are susceptible to oxidative stress. Photo-oxidation, mediated by PCI-0123 PDT, could potentially overload an already highly oxidized stressed state leading to cell death. The good tissue penetration depth achieved by PCI-0213 mediated PDT and the activation of melanosomes makes PDT of pigmented melanoma, for the first time, clinically relevant.

Animals↗

[Microsurgical repair of the defect with large area of bare tibia].

OBJECTIVE: To study the best method in repairing the defect with a large area of bare tibia. METHOD: Three hundred and twenty-two cases of large bare tibia defects were treated by three microsurgical methods. RESULTS: The bare tibiae in the 322 cases ranging from 12 x 3 cm to 24.0 x 3.5 cm were covered with myocutaneous flaps. Primary wound healing was achieved in 300 cases and delayed healing in 22 cases. All the cases were followed-up for 1 to 5 years. The function of the lower limbs was evidently improved. None of them was amputated due to deficit of skin coverage. The patients were all satisfied with the operative results. CONCLUSION: Transposition of the bridging skin flap pedicaled with the contralateral posterior tibial blood vessels can cover extensive bare tibia to avoid amputation. Other myocutaneous flaps from thoraco-umbilical, tensor muscle of fascia lata and scapular-lateral chest areas are good options too.

Accidents, Traffic↗

Decreased neuronal nitric oxide synthase-immunoreactivity and NADPH-diaphorase activity in rat pterygopalatine ganglion parasympathetic neurons and cerebrovascular innervation following long-term sympathectomy.

Neural control of tissue perfusion is mediated predominantly by sympathetic vasoconstrictor and, in some tissues, parasympathetic vasodilator systems. The objective of this study was to determine if changes occur within parasympathetic vasodilator neurons supplying cranial vessels after sympathectomy. Cranial sympathetic innervation was excised unilaterally, and 6 weeks later pterygopalatine ganglion neurons were evaluated for expression of neuronal nitric oxide synthase immunoreactivity (NOS-ir) and NADPH-diaphorase activity. Relative to the unoperated side, the number of parasympathetic neurons staining densely either for NOS or for NADPH-diaphorase decreased by 37% and 47%, respectively, whereas unstained somata increased by 51% and 48%. Internal carotid artery NOS-ir innervation density was reduced by 35%. These findings are consistent with a down-regulation of nitrergic properties in some cranial parasympathetic neurons after sympathectomy.

Animals↗

Decreased receptivity of pathway connective tissue to sympathetic nerve ingrowth in the developing rat.

Sympathetic axons can form atypical pathways to denervated orbital targets in neonatal rats but not in rats aged 30 or more days. The objective of this study was to determine if connective tissue pathways that carry sympathetic nerves lose their ability to sustain axonal sprouting during the early postnatal period. Regions of periorbital sheath known to contain large numbers of sympathetic axons that travel to distal orbital targets were excised from rats (sympathectomized 3 days previously) on postnatal days 6-7, 14-15, 30-31, and 48-49 and placed in anterior chambers of adult host rats. Tissues were removed 3, 6, or 10 days post-transplant and sympathetic ingrowth was analyzed by catecholamine histofluorescence in whole-mount or cryosectioned specimens. Connective tissue transplants from 6-15-day-old donors showed significant fiber ingrowth by 3 days in oculo, and innervation was maximal by 6 days. In contrast, sprouting into 30-49-day-old tissue was significantly slower, with most transplants lacking fibers at 3 days, and with small numbers of short fibers present at 6 days. We conclude that maturational changes occur in periorbital connective tissue pathways in the early postnatal period which make them less receptive to ingrowth by sympathetic nerves. The findings that connective tissue pathways are better substrates for sympathetic sprouting in the neonatal rat supports the view that developmental changes in these tissues are likely to contribute to the impaired reinnervation of orbital targets by contralateral neurons in juvenile and adult rats.

Aging↗

Comparative study on the inhibitory effect of recombinant FN polypeptide CH50 and CH56 on the metastasis of melanoma cells.

On the basis of preparation of anti-metastatic recombinant FN polypeptides, CH50 and CH56, we further studied the function of these polypeptides. The capacity of CH50 binding with melanoma cells (ED50 30 mM) was higher than that of CH56 (ED50 45 mM). Both of the polypeptides could significantly suppress the binding of melanoma B16 cells to laminin. There was no significant difference in the inhibitory effect between two polypeptides. In the experimental metastasis of melanoma cells, both of CH50 and CH56 could significantly inhibit the metastasis of the tumor cells, and reduce the number of lung metastasis by about 80%. Our results suggest that III-11 and ED-A repeats influenced, to some extent, the binding capacity of bifunctional-domain polypeptide to cells, but did not affect the inhibition of the polypeptide on the metastasis of melanoma cells. The presence and connection of cell I and Hep II domains are the elements which determine the ability of recombinant FN polypeptides to inhibit the metastasis of tumor cells.

Animals↗

Localization and efficacy analysis of the phototherapeutic lutetium texaphyrin (PCI-0123) in the murine EMT6 sarcoma model.

Lutetium texaphyrin (PCI-0123) is a pure, water-soluble photodynamic therapy (PDT) agent that is activated by tissue-penetrating far red light. The sensitizer is highly fluorescent and exhibits a strong, broad emission signal at 750 nm. In vitro cellular uptake studies revealed an increase in sensitizer retention with incubation time. Confocal laser scanning microscopy demonstrated that the intracellular localization site of PCI-0123 is the lysosomes. Ensuing illumination of the EMT6 cells led to lysosomal breakup, extensive cytoplasmic blebbing and subsequent cell death. Noninvasive spectral imaging analysis of PCI-0123 fluorescence depicted selective drug uptake, compared to surrounding normal tissue, in EMT6 mammary sarcomas syngeneic to BALB/c mice. The PCI-0123 PDT was shown to effectively treat the EMT6 murine sarcoma. Irradiation (732 nm light) 3 h postintravenous injection of 10 mumol PCI-0123 per kg gave 100% cures (no evidence of cancer), whereas light exposure at 5 h resulted in 75% cures. Hematoxylin and eosin histologic examination of photoirradiated tumors indicated apoptosis of the EMT6 neoplasms at early times post-PDT progressing, with time, to extensive necrotic areas. Gel electrophoresis of extracted photoirradiated tumors showed the typical apoptotic DNA ladder pattern that increased in intensity following PDT treatment.

Animals↗

C/EBP factor suppression of inhibition of type II secreted phospholipase A2 promoter in HepG2 cells: possible role of single-strand binding proteins.

We previously reported that the type II secreted phospholipase A2 (sPLA2) promoter from positions (-326 to +20) ([-326;+20] promoter) is negatively regulated by two adjacent regulatory elements, C (-210 to -176) and D (-247 to -210). This study examines in greater detail the way in which this negative regulation operates. Successive 5' deletions of the [-326;+20] type II sPLA2 promoter indicated that the region upstream of position -195 inhibits the transcription activity sixfold in HepG2 cells but not in HeLa cells. Although the whole [-326;-176] region decreased the activity of a heterologous thymidine kinase promoter, this effect was orientation and position sensitive. C/EBP beta, C/EBP alpha, and C/EBP delta, which bind to element C, prevented the inhibition of promoter activity. Electrophoretic mobility shift experiments identified the binding of NF1-like proteins to the [-225;-218] site, which overlaps an insulin response-like sequence, 5'-TGTTTTG-3'. This sequence bound a factor which also recognized the promoters of the apolipoproteins C-III and A-II. Substitutions preventing the binding of this factor or the NF1-like proteins did not increase the transcription activity, but substitution in the [-217;-204] sequence blocked the transcription inhibition. This sequence did not bind any double-strand binding factor, but its antisense strand is critical for the binding of single-strand binding proteins to the [-232;-191] region. We therefore suggest that these single-strand binding proteins are involved in the inhibitory mechanism.

Animals↗

[Treatment of malignant or aggressive bone tumors with microwave induced hyperthermia].

Limb-sparing procedures have been well established for dealing with malignant bone tumors. Unfortunately, these procedures have different problems. We used an alternative operation combined with microwave-induced hyperthemia to modify the surgical methods. Thermotherapy with microwave intracorporeal irradiation was used to treat 112 patients with bone tumors. In this series, 79 had malignant tumors and 33 aggressive benigh tumors. Postoperatively, immune therapy was carried out regularly. The patients immunologic functions were monitored by assay of the subpopulation of T cells, IL-2 and sIL-2R (soluble IL-2 receptor). Follow-up varied from 3 to 50 months (mean 23 month) s. Excluding 5 patients with malignancy in the vertebrae treated for palliation, 107 were evaluated by oncological and orthopedic criteria. 10 patients had local recurrence and required amputation. The remaining 97 had excellent local control. In 12 of the 74 patients with malignancy of the extremities, lung metastasis occurred 4 months to 2 years after surgery. Pathological fracture occurred at devitalized bone in 8 patients. In 29 out of 40 tumor-free cases followed for more than 2 years, the knee joints functioned properly with almost full range of motion. Single photon emission computered tomography (SPECT) study revealed revascularization of the devitalized tumor bearing bone segment could accomplish in one year or more. The immune state was improved after thermotherapy plus immunotherapy in the majority of patients. These results indicated that the use of microwave hyperthermia and adjuvant immunotherapy in the surgical treatment of bone tumors can be considered a definitive procedure, which is safe and well-tolerated.

Adolescent↗

[The effects of hyperinsulinmia and insulin-like growth factor-1 on hyperandrogenism in polycystic ovarian syndrome].

OBJECTIVE: To investigate the effects of insulin (INS) and insulin-like growth factor-1 (IGF-1) on ovarian androgen production in women with polycystic ovarian syndrome (PCOS). METHOD: 48 patients with PCOS were divided into two groups based on body mass index (BMI) (24 were obese BMI > 25 and 24 nonobese) (BMI < 25). 10 women with normal weight and menstruation were recruited as controls. The serum LH, FSH and testosterone (T) levels were determined. 40 cases were measured INS, IGF-1, androstenodine (A) and sex hormone binding globulin (SHBG) after a 3-hour oral glucose tolerance test (OGTT). Ovarian stroma obtained from 10 patients with PCOS and 5 healthy women were cultured in the medium with high INS concentration (30 ng/ml and 60 ng/ml). The IGF-1 and A levels were determined. RESULTS: Two groups of patients with PCOS demonstrated significant hyperinsulinmia (HI) and hyperandrogenism. Significantly higher IGF-1 level was observed in nonobese patients (P < 0.01). Insulin markedly stimulated release of IGF-1 (4 times, P < 0.01) and A (6 times, P < 0.05) in incubation of ovarian stroma obtained from PCOS patients. A positive correlation between IGF-1 and A was present in the culture medium (r = 0.679, P < 0.05). The serum levels of SHBG were significantly lower in PCOS patients. CONCLUSION: The findings suggest that the women with PCOS have HI, since INS may play an important role in hyperandrogenism by upregulating IGF-1 system in ovary.

Adult↗

Antisense ribosomes: rRNA as a vehicle for antisense RNAs.

Although rRNA has a conserved core structure, its size varies by more than 2000 bases between eubacteria and vertebrates, mostly due to the size variation of discrete variable regions. Previous studies have shown that insertion of foreign sequences into some of these variable regions has little effect on rRNA function. These properties make rRNA a potentially very advantageous vehicle to carry other RNA moieties with biological activity, such as "antisense RNAs." We have explored this possibility by inserting antisense RNAs targeted against one essential and two nonessential genes into a site within a variable region in the Tetrahymena thermophila large subunit rRNA gene. Expression of each of the three genes tested can be drastically reduced or eliminated in transformed T. thermophila lines containing these altered rRNAs. In addition, we found that only antisense rRNAs containing RNA sequences complementary to the 5' untranslated region of the targeted mRNA were effective. Lines containing antisense rRNAs targeted against either of the nonessential genes grow well, indicating that the altered rRNAs fulfill their functions within the ribosome. Since functional rRNA is extremely abundant and stable and comes into direct contact with translated mRNAs, it may prove to be an unparalleled vehicle for enhancing the activity of functional RNAs that act on mRNAs.

Bacterial Proteins↗

Sympathetic nerve trajectories to rat orbital targets: role of connective tissue pathways.

The trajectories of sympathetic nerves projecting to orbital targets were determined in adult rats with intact innervation and following acute sympathetic denervation, neonatal unilateral superior cervical ganglionectomy, or unilateral ganglionectomy on postnatal day 30. Sympathetic nerves were identified by using immunofluorescence for the noradrenergic transmitter enzyme dopamine beta-hydroxylase and by using catecholamine histofluorescence. In rats with intact innervation, sympathetic fibers travel to the orbit in association with the abducens, trochlear, and Vidian nerves. Within the retroorbital and retroocular connective tissue, the fibers redistribute to become associated with sensory-nerve branches of the trigeminal nerve, the orbital vasculature, and the periorbital sheath. Fibers reach their targets by traversing variable amounts of connective tissue of the periorbitum, the orbital septa, and the striated muscle epimysia. Following neonatal ganglionectomy, intracranial fibers of contralateral origin enter the orbit by traveling through connective tissue of the optic nerve meninges and lining the anterior lacerated foramen. These fibers travel independent of the trochlear, abducens, and Vidian nerves, but, otherwise, they use the same orbital pathways as those employed in the intact animal. In animals ganglionectomized on postnatal day 30, fibers enter the posterior portion of the orbit primarily via the optic foramen; they travel only short distances and end blindly in the periorbital sheath. These findings indicate that fascial structures are a major component of the pathways that guide sympathetic fibers to their appropriate targets both in normal development and during reinnervation following neonatal ganglionectomy. Because orbital connective tissues are termination sites of abortive fiber sprouting in older rats, developmental changes in the properties of these tissues may contribute to the absence of pathway formation in the mature animal.

Adrenergic Fibers↗

Effect of physical constraints on the mechanisms of membrane fusion: bolaform lipid vesicles as model systems.

Bolaform lipid vesicles were used to study the effect of physical constraints on membrane fusion. In these vesicles the membrane is organized in a single monolayer, because of the presence of covalent bonds in its middle plane. Therefore, the formation of fusion intermediates is subject to higher energy barriers and greater geometrical constraints than is usual in bilayer membranes. Bolaform lipids were extracted from the thermophilic archaeon Sulfolobus solfataricus. These lipids can be divided into two classes, the monosubstituted molecules, in which one of the polar heads is glycerol, and the bisubstituted molecules, endowed with two complex polar heads. The fusion process in vesicles composed of different mixtures of monosubstituted/bisubstituted molecules was studied by means of fluorescence techniques. Ca2+ or poly(ethylene glycol) was employed as a fusogenic agent. We found that fusion of such constrained membranes is still possible, provided that molecules able to mediate a structural rearrangement of the membrane are present. This condition is fulfilled by monosubstituted molecules, which are able to partition the glycerol headgroup in the apolar moiety. In addition, the presence of traces (approximately 5%) of the monopolar compound diphytanylglycerol is an important factor for fusion to occur. On the contrary, vesicles formed by bisubstituted molecules are unable to fuse, irrespective of the fusogen employed.

Cell Membrane↗

Psychological long-term effects of sterilization on anxiety and depression.

Two-thousand cases (500 women and 500 men with sterilization; 500 women and 500 men without sterilization) in two counties and two cities in Sichuan, China, were investigated between 15 September 1992 and 30 April 1993. Information was obtained about their age, parity, marriage, contraceptives, social behavior, and present psychological characteristics. CES-D, SAS, and E.P.Q. scales were used to assess the depressive symptomatology, anxiety, and personality of the subjects. The results revealed that sterilization psychologically affected the depressive symptoms and anxiety under the neutral personality. In addition, depressive symptoms and anxiety were related to the subjects' age, educational level, income, operative doctors' attitudes to sterilization acceptors, and subjects' understanding of the sterilization.

Adult↗