[Advances in the construction of risk prediction models for chronic kidney failure].

Chronic kidney disease (CKD) is a major public health problem worldwide. When CKD patients progress to the stage of kidney failure, kidney replacement therapy (KRT) or conservative treatment (palliative or non-dialysis treatment) will be needed. The risk prediction models of chronic kidney failure have been developed in recent years. These models, focusing on demographic indicators, clinical indicators, and laboratory data, are used to predict the likelihood of progression to kidney failure and requiring KRT. This article will retrieve prediction models for chronic kidney failure as an outcome, demonstrate the current research progress, and hope that it may be helpful for the strategies of preventing chronic kidney failure.

[Clinical application analysis of a method for locating scalp projection of intracranial lesions based on neuroimaging].

Objective: To explore the feasibility of a method based on neuroimaging and surface markers for locating scalp projection of intracranial lesions. Methods: The clinical data of 46 patients who were used 'double-circle method' for locating scalp projection of intracranial lesions at Department of Neurosurgery,the First Affiliated Hospital of Xiamen University from January to June 2021 were retrospective analyzed. All patients with 2 electrodes(artificial fiducials) randomly attached to scalp had been examed thin-layer brain CT. The distances from the center of each fiducial to the root of the nose and tragus were measured through the images. A compass was used to draw two arcs with the root of nose and the tragus as the center and the pre-measured distance as the radius on patient's scalp. Then two arcs' intersection on the scalp was the fiducial. The method was named 'double-circle method'. Two neurosurgeons were arranged to perform fiducial identification with double-circle method, and record the error between the result and the actual fiducial point.Independent sample t test was used for data comparison, and Kappa test was used to analysis the inter-group consistency. Results: Ninety-two fiducial points of 46 patients were collected. Time consuming of doctor A was (8.1±2.3) minutes(range:5 to 15 minutes)and doctor B was (8.9±3.5) minutes(range:4 to 17 minutes).The positioning error from the doctor A was (4.4±2.4)mm(range:0 to 12 mm) and doctor B was(4.2±2.6) mm(range:0 to 14 mm)(t=-0.575,P=0.567),the difference was not statistically significant. The Kappa value of the consistency test of error between two doctors was 0.517(P=0.001).The consistency was moderate.Eight patients used 'double-circle method' and neuronavigation for locating scalp projection of intracranial lesions at the same time. The diameter of the lesions was (3.8±0.9)cm (range: 2.6 to 5.1 cm), and the positioning error of the 'double-circle method' and navigation was (4.0±1.9) mm(range: 1 to 6 mm), and all patients were confirmed to be accurately located during surgery. Conclusion: 'Double-circle method' is a simple,convenient and accurate way in locating intracranial lesions and has certain clinical significance.

[Role of personalized 3D printing in brain protection after decompressive craniectomy].

Objective: To explore the application value of personalized three-dimensional (3D) printed protective cap in brain protection after decompressive craniectomy (DC). Methods: Fourty-five patients who underwent DC from January 2021 to October 2021 were selected, including 26 males and 19 females, aged 5-73 (50±13) years old. The brain CT data were imported into 3D Slicer software to rebuild the protective cap through 3D printing. The cap was worn on the head of the patient, thereby preventing secondary braindamage. The follow-up results were compared with 53 patients without protective capduring the same period. Results: There were no statistically significant differences in age, skull defect location and follow-up time between the two groups (all P>0.05).Among 45 patients, 47 brain protective caps (2 cases with bilateral skull defects) were successfully designed. The time for image post-processingand 3D printing was (21.2±6.0) min and (62.4±8.3) min, respectively. There were 6 cases of low compliance, 9 cases of moderate compliance, 32 cases of high compliance, respectively. Six cases with low conformity were redesigned and printed, 2 of 9 cases with moderate conformity were redesigned and printed, and the remaining 7 cases reached high compliance after grinding and packaging. In the current study, 45 patients with brain protective caps were followed up for 3 months, and no secondary brain injury occurred. However, among 53 patients without brain protective caps during the same period, 4 patients had secondary accidental brain compression. The incidence of injury was 7.5 %, and the difference was statistically significant (P<0.001). Conclusion: Brain protective cap designed based on cranial CT and 3D printing can be used in patients with skull defects to protect the brain tissue from secondary crush damage and has certain clinical value.

Long non-coding RNA DANCR induces chondrogenesis by regulating the miR-1275/MMP-13 axis in synovial fluid-derived mesenchymal stem cells.

OBJECTIVE: The aim of this study was to examine the role of lncRNA-differentiation antagonizing non-protein coding RNA (DANCR) and its underlying mechanisms in chondrogenesis, more specifically in synovial fluid-derived mesenchymal stem cell (SFMSCs). MATERIALS AND METHODS: The expression levels of DANCR in SFMSCs were measured by qRT-PCR. Luciferase reporter assay and RIP assay were used to investigate the direct target of DANCR and miR-1275 in SFMSCs. The expression of matrix metallopeptidase 13 (MMP13, also known as chondrogenic marker) protein was examined by Western blot. Cell proliferation was analyzed by Cell Counting Kit-8 (CCK-8) assay, while chondrogenic differentiation was explored by sGAG assay. RESULTS: Our data indicated that DANCR can promote SFMSCs proliferation and chondrogenesis. In addition, miR-1275 was indicated as a direct target of DANCR. MiR-1275 was negatively regulated by DANCR via competing endogenous RNA (ceRNA) mechanism. Moreover, our data revealed that miR-1275 could bind to MMP13 and regulate its expression. CONCLUSIONS: Our findings suggested that DANCR was involved in SFMSCs proliferation and chondrogenesis. Mechanistically, DANCR functions as a sponge RNA for miR-1275 that regulates the expression of target gene MMP13. These data provide a therapeutic option for Osteoarthritis (OA).

[Clinical study on asthma and aspirin asthma affecting chronic rhinosinusitis].

Objective:The aim of this study is to observe the effects of asthma and aspirin asthma on chronic rhinosinusitis and to explore the corresponding clinical value. Method: Eighty-six patients with CRS and asthma who were treated in the outpatient clinic during March 2015 to January 2018 were divided into asthma group(52 cases) and aspirin asthma group(34 cases) according to asthma and aspirin asthma. The clinical symptoms of the two groups were analyzed by symptomatic VAS score, Lund-Mackay score of sinus CT, and Lund-Kennedy score by nasal endoscopy.The scores of the two groups were compared under different lung function. Enzyme-linked immunosorbent assay the levels of inflammatory markers IL-5,IL-17,IFN-γ and TNF-α in the sinus secretions of the two groups were detected.Result:There were no significant differences in age, gender, smoking history, allergy history, surgical history and course of disease between the two groups(P<0.05), suggesting that the data were comparable. The sinus CT results showed that compared with the aspirin asthma group, the asthmatic group had irregular turbinates and a large turbinate,as shown in Figure 1. There were significant differences between the two groups in VAS score,Lund-Mackay score of sinus CT and Lund-Kennedy score by nasal endoscopy.The difference was statistically significant(P<0.05). And the forehead and/or facial pain or pain in the symptomatic VAS score(P<0.05), the Lund-Mackay score of the sinus CT(P<0.05),and intranasal.The difference in the Lund-Kennedy score(P<0.05) was statistically significant.There were significant differences in the distribution of lung function levels between the two groups of patients with mild airway obstructive respiratory dysfunction and pulmonary ventilation obstructive disorder(P<0.05).The average levels of IL-5,IL-17,IFN-γ and TNF-α in the aspirin asthma group were significantly lower than those in the asthma group(P<0.05).Conclusion:Aspirin-induced CRS produces asthma symptoms more severely than traditional asthma symptoms, but the induced local inflammatory response is relatively weak, and the mechanism may be closely related to IL-5,IL-17,IFN-γ and TNF-α levels.

[Clonal evolution and clinical significance of trisomy 8 in acquired bone marrow failure].

Objective: To analyze clonal evolution and clinical significance of trisomy 8 in patients with acquired bone marrow failure. Methods: The clinical data of 63 patients with acquired bone marrow failure accompanied with isolated trisomy 8 (+8) from June 2011 to September 2018 were analyzed retrospectively, the clonal evolution patterns and relationship with immmunosuppressive therapy were summarized. Results: Totally 24 male and 39 female patients were enrolled, including 39 patients with aplastic anemia (AA) and 24 patients with relatively low-risk myelodysplastic syndrome (MDS) . Mean size of+8 clone in MDS patients[65% (15%-100%) ]was higher than that of AA patients[25% (4.8%-100%) , z=3.48, P=0.001]. The patients were was divided into three groups (<30%, 30%-<50%,and ≥50%) according to the proportion of+8 clone. There was significant difference among the three groups between AA[<30%:55.6% (20/36) ; 30-50%: 22.2% (8/36) ; ≥50%22.2% (8/36) ]and MDS patients[<30%:19.0% (4/21) ; 30%-<50%:19.0% (4/21) ; ≥50%61.9% (13/21) ] (P=0.007) . The proportion of AA patients with+8 clone <30% was significantly higher than that of MDS patients (P=0.002) ; and the proportion of AA patients with+8 clone ≥50%was significantly lower than that of MDS patients (P=0.002) . The median age of AA and MDS patients was respectively 28 (7-61) years old and 48.5 (16-72) years old. Moreover, there was no correlation between age and+8 clone size in AA or MDS (r(s)=0.109, P=0.125; r(s)=-0.022, P=0.924, respectively) . There was statistical difference in total iron binding capacity, transferrin and erythropoietin between high and low clone group of AA patients (P=0.016, P=0.046, P=0.012, respectively) , but no significant difference in MDS patients. The immunosuppressive therapy (IST) efficacy of AA and MDS patients was respectively 66.7% and 43.8% (P=0.125) . Comparing with initial clone size (27.3%) , the +8 clone size (45%) of AA patients was increased 1-2 year after IST, but no statistical difference (z=0.83, P=0.272) . Consistently, there was no significant change between initial clone size (72.5%) and 1-2 year clone size (70.5%) after IST in MDS patients. There was no significant difference in IST efficient rate between +8 clone size expansion and decline group of in AA patients at 0.5-<1, 1-2 and>2 years after IST. We found four dynamic evolution patterns of +8 clone, which were clone persistence (45%) , clone disappearance (30%) , clone emergence (10%) and clone recurrence (15%) . Conclusions: AA patients had a low clone burden, while MDS patients had a high burden of +8 clone. The +8 clone of AA patients didn't significantly expanded after IST, and the changes of +8 clone also had no effect on IST response.

[Effects and related mechanism of overexpression of human thioredoxin on the inflammatory response in mice with viral myocarditis].

Objective: To observe the effects of recombinant adenovirus with human thioredoxin (hTRX) on the inflammatory response in mice with viral myocarditis and explore the related mechanism. Methods: Sixty Balb/c male mice were randomly divided into control group, myocarditis group, and hTRX group according to the random number table (n=20 each group). The myocarditis group and hTRX group were injected with 100 TCID(50) Coxackie virus B3 (0.1 ml) in the abdomen and control group were injected with saline. Two days before the viral injection, the hTRX group were injected with recombinant adenovirus vector coding the human thioredoxin gene by pericardial puncture and the control group and myocarditis group were injected with recombinant adenovirus vector without coding gene by pericardial puncture, all these mice were killed and hearts were removed 7 days later. The morphology of myocardial tissue in each group was detected by HE staining and the ultrastructure changes by electron microscope. The protein expressions of tumor necrosis factor (TNF)-α, interleukin (IL)-1ß and NF-κB were detected by ELISA and Western blot. Immunohistochemical staining was performed to observe the protein expression levels of thioredoxin. Results: Necrosis of myocardial cells and a small amount of cell infiltration were found in the myocarditis group and necrosis and cell infiltration were significantly reduced in the hTRX group and no myocardial lesion was found in control group on HE stained sections. Electron microscope examination evidenced cell swelling and dissolved myofilament, vacuoles degeneration in mitochondria in the myocarditis group. These changes were significantly reduced in the hTRX group. There was no myocardial lesion in control group. The protein expression of TNF-α, IL-1ß and NF-κB were significantly upregulated in myocarditis group than in control group (all P<0.01). The protein expression of TNF-α, IL-1ß and NF-κB were significantly downregulated in hTRX group than in myocarditis group (all P<0.01). Immunohistochemical staining showed that protein expression of hTRX was higher in hTRX group than in myocarditis group (P<0.01). Conclusion: Recombinant adenovirus hTRX can attenuate cardiac injury in mice with acute myocarditis via inhibiting the inflammatory response and downregulating the expression of TNF-α, IL-1ß and NF-κB.

Selenoprotein P and Yunnan endemic sudden cardiac death--an ecological study.

The aim of the present study was to explore the role of selenoprotein P (SePP) in the etiology of the endemic sudden cardiac death in Yunnan, China. The levels of SePP of 124 subjects and glutathione peroxidase (GPx) of 119 subjects were measured. The subjects were from the old and new endemic areas and non-endemic areas. The levels of SePP and GPx of the subjects of the old endemic area were significantly higher than those of the subjects of the new endemic area and the non-endemic areas, respectively. The Pearson's correlation among SePP, GPx, and the number of the incident cases of the disease were statistically significant. These correlations show that there is an inverse relationship among the number of patients and the levels of SePP (r = - 0.9800, P = 0.0200) and GPx (r = - 0.961, P = 0.009). The results show that selenium deficiency might play an important role in the incidence of the disease.

Analysis of glutathione peroxidase 1 gene polymorphism and Keshan disease in Heilongjiang Province, China.

Keshan disease (KD) is an endemic cardiomyopathy associated with selenium deficiency. Recent studies indicate that glutathione peroxidase 1 (GPx1) mutation decreases GPx activity in myocardial cells and increases the risk of KD. To further clarify the correlation between GPx1 polymorphism and KD, we analyzed GPx1 polymorphism, blood selenium levels and GPx activity in KD patients and healthy controls in Heilongjiang Province. Four and 24 new mutation loci in the promoter and the exon region, respectively, of the GPx1 gene were found in the subjects, in contrast with the previously reported loci. There were no significant differences in the mutation frequency of these loci between the KD group and controls (chi-square test; P > 0.05). However, the mutation frequency of exon 474 was higher in the KD group (7/36) than in controls (2/41), and GPx activity was lower in the mutation group (90.475 ± 23.757 U/L) than in the non-mutation group (93.947 ± 17.463 U/L). Further investigation is necessary to clarify a possible causality between GPx1 exon 474 mutation and KD.

A comparison study of detecting gold nanorods in living cells with confocal reflectance microscopy and two-photon fluorescence microscopy.

Two-photon fluorescence microscopy and confocal reflectance microscopy were compared to detect intracellular gold nanorods in rat basophilic leukaemia cells. The two-photon photoluminescence images of gold nanorods were acquired by an 800 nm fs laser with the power of milliwatts. The advantages of the obtained two-photon photoluminescence images are high spatial resolution and reduced background. However, a remarkable photothermal effect on cells was seen after 30 times continuous scanning of the femto-second laser, potentially affecting the subcellular localization pattern of the nanorods. In the case of confocal reflectance microscopy the images of gold nanorods can be obtained with the power of light source as low as microwatts, thus avoiding the photothermal effect, but the resolution of such images is reduced. We have noted that confocal reflectance images of cellular gold nanorods achieved with 50 microW 800 nm fs have a relatively poor resolution, whereas the 50 microW 488 nm CW laser can acquire reasonably satisfactory 3D reflectance images with improved resolution because of its shorter wavelength. Therefore, confocal reflectance microscopy may also be a suitable means to image intracellular gold nanorods with the advantage of reduced photothermal effect.

Short axis contact in the chaining of ellipsoidal particles of polar molecule dominated electrorheological fluid.

We simulate the effect of the chaining direction of ellipsoidal particles of polar molecule dominated electrorheological (PM-ER) fluids using commercially available COMSOL Multiphysics® software for the distribution of electric field and the total electrostatic energy. It is proved that adding ferroelectric materials to the channels parallel to the short axis would make the short axis parallel to the field direction when the ellipsoidal particles are chained under an electric field. According to our simulation, while the concentration of the channels stays constant, the greater the dielectric constant of the inserted material, the stronger the maximum local electric field will be.

Saturated orientational polarization of polar molecules in giant electrorheological fluids.

Many researches on polar-molecular electrorheological (PMER) fluids with giant electrorheological effects were reported in recent years. The particles of PMER fluids (PMER particles) are known to have a dielectric core with high dielectric constant and a shell of polar molecules. Our calculation of local electric fields using the finite element approach shows that the local electric field can cause an orientational polarization of the polar molecules. The saturation of the orientational polarization occurs on the outer shells of two nearby PMER particles. Then, it causes the strong outer shell-outer shell interaction between the two particles, and this kind of interaction is just responsible for the giant electrorheological effect. It is further realized that the PMER effect is mainly due to the interaction of the tail-head connected polar molecules within the two outer shells between the two PMER particles. Our theoretical results of static yield stresses are shown to be in excellent agreement with the reported experimental data by several groups. For general PMER fluids, the calculated static yield stress is nearly proportional to R(x-1). When h/R, the ratio between the thickness of shells and radius of PMER particles, changes from 0.05 to 0.5, the index x changes accordingly from 0.64 to 0.51. It is also found that particles with thinner thickness h and smaller radius R have larger electrorheological effects until the static yield stress shows a peak when R reaches about 10 nm.

On the Lorentz local electric field in soft-matter systems.

In electric-field-responsive soft-matter systems, the suspended particles respond to the Lorentz local field (LLF), yielding abundant important phenomena. Even though the particles can easily rotate, the LLF was conventionally adopted as a quantity that is independent of rotations in the literature. In contrast, here we design an experiment to measure the LLF between two metallic spheres, one of which is rotating, and report a rotation-driven reduction. Excellent agreement between our experiment and theory reveals the role of the relaxation of dipole moments. Its relevance to biophysics, colloidal physics, and nonlinear physics is also discussed.

Structure of electrorheological fluids under an electric field and a shear flow: experiment and computer simulation.

It is known that macroscopic properties of colloidal suspensions are often determined by the microstructure of the particles in the suspensions, depending on the interparticle, Brownian, and hydrodynamic (if any) forces. We take electrorheological (ER) fluids as an example. By using a computer simulation and an experimental approach, we investigate the structure of ER fluids subjected to both an electric field and a shear flow. The microstructure evolution from random structure, to chains, and then to stable lamellar patterns, observed in the experiments, agrees very well with that obtained in the simulations. It is shown that the formation of such lamellar patterns originates from the difference between the dipole moment induced in the particles suspended in the ER fluids without shear and the one with shear. The results on the relaxation process of structural formation and the internal structure of layers are also presented. Thus, it seems possible to achieve various structures and hence desired macroscopic properties of colloidal suspensions by adjusting external fields and, simultaneously, a shear flow.

Effect of low power laser irradiation on disconnecting the membrane-attached hemoglobin from erythrocyte membrane.

In our previous study we found that low power laser irradiation improved the erythrocyte deformability, but the mechanism is unclear. The membrane-attached hemoglobin (Hbm) may be one of the determining factors for the erythrocyte deformability. We report here for the first time, that laser irradiation can reduce the Hbm contents in pig's erythrocytes, providing the explanation for the improvement of erythrocyte deformability. The decrease of the Hbm was proportional to the irradiation dose, but the relative change of Hbm was saturated around 35%. The 532 nm laser was more efficient at lowering Hbm than the 632.8 nm laser, consistent with the absorption spectrum of Hbm.

A comparative study of 632.8 and 532 nm laser irradiation on some rheological factors in human blood in vitro.

The effects of laser irradiation with 632.8 and 532 nm on rheological properties of blood were comparatively studied in vitro. Under the irradiation condition of 30 mW, laser irradiation of blood samples using a spot diameter of 5 mm with each laser, showed promising results in the modulation of hemorheological properties. When blood samples from patients with abnormally high values of erythrocyte sedimentation rate (ESR) were irradiated, the values of ESR were lowered statistically by either of the 632.8 or 532 nm lasers. The laser irradiation reduced blood viscosities at different shear rates (10-110 S(-1)) for the hyper-viscosity blood samples. Laser irradiation increased the electrophoretic mobility (EPM) of erythrocytes when the values of the sample's EPM were abnormally slow. The erythrocyte deformability was enhanced by laser irradiation when the deformability of the sample from the patients was originally poor. For verifying the improvement of laser irradiation on erythrocyte deformability, the typical erythrocyte samples with poor deformability were produced by the pre-treatment of the erythrocytes with Ca(2+). The deformability of these erythrocyte samples was also improved after laser irradiation. These results suggest that membrane-bound hemoglobin (Hbm) might be the initial site of the interaction, since Hbm is the main cause of poor deformability when erythrocytes were treated with Ca(2+). In all experiments including ESR, blood viscosity, EPM and erythrocyte deformability, the 532 nm laser demonstrated more efficient effects on modulating rheological properties than 632.8 nm laser. This wavelength effect is consistent with the absorption spectrum of hemoglobin, reflecting that hemoglobin may be one of the action targets under laser irradiation.

Antisense RNA gene therapy for studying and modulating biological processes.

Agents that produce their effects through an antisense mechanism offer the possibility of developing highly specific alternatives to traditional pharmacological antagonists, thereby providing a novel class of therapeutic agents, ones which act at the level of gene expression. Among the antisense compounds, antisense RNA produced intracellularly by an expression vector has been used extensively in the past several years. This review considers the advantages of the antisense RNA approach over the use of antisense oligodeoxynucleotides, the different means by which one may deliver and produce antisense RNA inside cells, and the experimental criteria one should use to ascertain whether the antisense RNA is acting through a true antisense mechanism. Its major emphasis is on exploring the potential therapeutic use of antisense RNA in several areas of medicine. For example, in the field of oncology antisense RNA has been used to inhibit several different target proteins, such as growth factors, growth factor receptors, proteins responsible for the invasive potential of tumor cells and proteins directly involved in cell cycle progression. In particular, a detailed discussion is presented on the possibility of selectively inhibiting the growth of tumor cells by using antisense RNA expression vectors directed to the individual calmodulin transcripts. Detailed consideration is also provided on the development and potential therapeutic applications of antisense RNA vectors targeted to the D2 dopamine receptor subtype. Studies are also summarized in which antisense RNA has been used to develop more effective therapies for infections with certain viruses such as the human immunodeficiency virus and the virus of hepatitis B, and data are reviewed suggesting new approaches to reduce elevated blood pressure using antisense RNA directed to proteins and receptors from the renin-angiotensin system. Finally, we outline some of the problems which the studies so far have yielded and some outstanding questions which remain to be answered in order to develop further antisense RNA vectors as therapeutic agents.

Cytosolic-Ca2+ and coxsackievirus B3-induced apoptosis in cultured cardiomyocytes of rats.

AIM: To explore the role of cytosolic free calcium ([Ca2+]i) in apoptosis induced by coxsackievirus B3 (CVB3) in cultured cardiomyocytes of rats. METHODS: Primary cultured cardiomyocyte was prepared from Wistar rats ages 2-3 d. The apoptosis in cardiomyocyte was determined by terminated deoxynucleotide transferase directed d-UTP nick and end labeling (TUNEL) method, and the apoptosis was observed under a transmission electron microscope. [Ca2+]i in single cardiomyocyte loaded with Fluo 3-AM was measured by confocal microsorope. RESULTS: (1) The concentration of CVB3 in the medium reached the peak at 24 h after CVB3 infection. (2) The apoptotic cells were not found in CVB3-infected cardiomyocyte in first 10 h, but amounted to 5% at 17 h, 60% at 24 h, and 90% at 36 h. (3) The peak value of [Ca2+]i elevation reached at 17 h after CVB3 infection (P < 0.01). (4) The characteristics of apoptosis was also seen by transmission electron microscope. CONCLUSION: CVB3 induced the apoptosis in cultured cardiomyocyte, and [Ca2+]i mobilization was involved in the signal transduction process in apoptosis cells, and played an important role especially in the early stage of apoptosis induced by CVB3.

Isolation and characterization of a new gene, sre, which encodes a GATA-type regulatory protein that controls iron transport in Neurospora crassa.

Multiple GATA factors - regulatory proteins with consensus zinc finger motifs that bind to DNA elements containing a GATA core sequence - exist in the filamentous fungus Neurospora crassa. One GATA factor, NIT2. controls nitrogen metabolism, whereas two others, WC-1 and WC-2, regulate genes responsive to blue light induction. A gene encoding a new GATA factor, named SRE, was isolated from Neurospora using a PCR-mediated method. Sequence analysis of the new GATA factor gene revealed an ORF specifying 587 amino acids, which is interrupted by two small introns. Unlike all previously known Neurospora GATA factors, which possess a single zinc-finger DNA-binding motif, SRE contains two GATA-type zinc fingers. The deduced amino acid sequence of SRE shows significant similarity to URBSI of Ustilago and SREP of Penicillium. A loss-of-function mutation was created by the RIP procedure. Analysis of sre+ and sre- strains revealed that SRE acts as a negative regulator of iron uptake in Neurospora by controlling the synthesis of siderophores. Siderophore biosynthesis is repressed by high iron concentrations in the wild-type strain but not in sre- mutant cells. The sre promoter contains a number of GATA sequences; however, expression of sre mRNA occurs in a constitutive fashion and is not regulated by the concentration of iron available to the cells.

D2 dopamine antisense RNA expression vector, unlike haloperidol, produces long-term inhibition of D2 dopamine-mediated behaviors without causing Up-regulation of D2 dopamine receptors.

Long-term inhibition of D2 dopamine receptors using classic D2 dopamine receptor antagonists such as haloperidol often causes a compensatory up-regulation of the D2 dopamine receptors. We investigated whether the long-term inhibition of D2 dopamine receptors using an eukaryotic expression vector housing a cDNA sequence encoding an antisense RNA directed to the D2 dopamine receptor transcript (D2 antisense vector) would also produce up-regulation of the D2 receptors. Single, bilateral injections of the D2 antisense vector into the corpora striata of mice inhibited the stereotypy induced by acute challenge injections with the D2/D3 dopamine receptor agonist quinpirole but did not inhibit the grooming induced by acute challenge injections with the D1 agonist SKF 38393. Similar treatment with the D2 antisense vector produced a long-term (>1 month) cataleptic response without producing tolerance to challenge injections with haloperidol. By contrast, catalepsy induced by a single injection of haloperidol lasted only approximately 2 days, and tolerance developed to its effects after long-term treatment. Repeated treatment of mice with haloperidol resulted in an inhibition of apomorphine-induced climbing behavior throughout the time of treatment with haloperidol, but the climbing behavior markedly increased to levels significantly higher than that of the control mice immediately after withdrawal from haloperidol treatment. This increased climbing was accompanied by increased levels of D2 dopamine receptors in the striatum. By contrast, single, bilateral intrastriatal injections of the D2 antisense vector significantly inhibited apomorphine-induced climbing for approximately 30 days but failed to increase the climbing behavior or the levels of D2 dopamine receptors in striatum over those of the control values. These results suggest that a single injection of a D2 antisense RNA expression vector into mouse striatum produces specific, long-term inhibition of D2 dopamine receptor behaviors without causing a compensatory increase in the levels or function of D2 dopamine receptors.