[Research advances on the regulatory mechanism of sweat secretion ion channels of eccrine sweat glands].

Sweat glands are widely distributed in human skin, among which eccrine sweat glands play major roles in heat dissipation and sweat secretion. Sweat secretion is mainly regulated by nervous system and includes two processes of secretion of secretory coil and reabsorption of sweat duct, involving various ion channels and proteins such as calcium ion channel, potassium ion channel, sodium-potassium-chloride co-transporter 1, Best2 protein, aquaporin 5, cystic fibrosis transmembrane conductance regulator, and epithelial sodium ion channel. This paper reviews the nerve conduction system and various ion channels involved in sweat secretion of exocrine sweat glands in order to provide a theoretical basis for the study of regeneration, repair, and transformation of stem cells.

Carbon dots as fluorescent probes for detection of VB12 based on the inner filter effect.

In this study, we constructed a new fluorescent sensing for VB12 and investigated the mechanism of vitamin B12 (VB12) quenching fluorescence of carbon dots (CDs). The fluorescence suppression is attributed to the inner filter effect (IFE) because of the overlap between UV-vis absorption spectrum of VB12 and emission/excitation spectra of CDs. This CDs-based sensor provides obvious advantages of simplicity, convenience, rapid response, high selectivity and sensitivity, which has potential application for the detection of VB12 in the medical and food industry.

Iron-embedded C2N monolayer: a promising low-cost and high-activity single-atom catalyst for CO oxidation.

An Fe-embedded C2N monolayer as a promising single-atom catalyst for CO oxidation by O2 has been investigated based on first-principles calculations. It is found that the single Fe atom can be strongly trapped in the cavity of the C2N monolayer with a large adsorption energy of 4.55 eV and a high diffusion barrier of at least 3.00 eV to leave the cavity, indicating that Fe should exist in the isolated single-atom form. Due to the localized metal 3d orbitals near the Fermi level, the embedded Fe single-atom catalyst has a high chemical activity for the adsorption of CO and O2 molecules. CO oxidation by O2 on the catalyst would proceed via a two-step mechanism. The first step of the CO oxidation reaction has been studied via the Langmuir-Hinshelwood and Eley-Rideal mechanisms with energy barriers of 0.46 and 0.65 eV, respectively. The second step of the CO oxidation reaction follows the Eley-Rideal mechanism with a much smaller energy barrier of 0.24 eV. For both the steps, the CO2 molecules produced are weakly adsorbed on the substrates, suggesting that the proposed catalyst will not be poisoned by the generated CO2. Our results indicate that the Fe-embedded C2N monolayer is a promising single-atom catalyst for CO oxidation by O2 at low temperatures.

Solid Lipid Nanoparticles as Non-Viral Vectors for Gene Transfection in a Cell Model of Fabry Disease.

Here, we demonstrate the ability of solid lipid nanoparticle-based non-viral vectors to increase the α-galactosidase A levels of the IMFE1 cell line, an in vitro model for target cells in Fabry disease. For this purpose, vectors containing the pR-M10-αGal A plasmid, which encodes the α-galactosidase A enzyme, were prepared; the in vitro transfection efficacy was studied in IMFE1 cells, and the results were confirmed by RT-PCR. The cellular uptake of the vectors, intracellular disposition of the plasmid, and probable endocytosis pathways of the nanoparticles were also analyzed. The vectors used for the studies carried protamine (P-DNA-SLN), dextran and protamine (D-P-DNA-SLN), or hyaluronic acid of two different molecular weights and protamine (HA150-P-DNA-SLN or HA500-P-DNA-SLN). The new formulations, which presented a particle size in the range of nanometers (from 218 nm to 348 nm) and a positive superficial charge, were able to increase α-galactosidase A activity up to 4-fold in comparison to non treated IMFE1 cells. The most efficient vectors were those that included HA, and no differences due to changes in the molecular weight of HA were detected. The observed lack of colocalization with each of the four different Nile Red-labeled vectors and transferrin or cholera toxin appears to indicate that clathrin- and caveolae-independent pathways may be involved in their cellular uptake. Additionally, colocalization with LysoTracker indicated that the formulations were exposed to lysosomal activity, which may be responsible for the release of the plasmid from the vector. In conclusion, we reveal the potential of SLN-based vectors to efficiently transfect an immortalized Fabry patient cell line.

Differentiation-inducing effects of betamethasone on human glioma cell line U251.

We studied the differentiation-inducing effect of beta-methasone on human glioma cell line U251 cultured in vitro, and the underlying mechanism. U251 cells were divided into two groups: control group cells, cultured in Dulbecco's Modified Eagle's medium containing 10% fetal bovine serum; and medication group cells, treated with 15 µM betamethasone. Morphological cell changes were observed by inverted microscope, cell cycle changes were ascertained by flow cytometry, and vimentin expression was checked by immunocytochemistry. The expression levels of extracellular signal-regulated protein ki-nase (ERK), phosphorylated ERK (pERK), and glial fibrillary acidic protein (GFAP) were assessed by western blot. Compared with the control group, U251 cell processes increased significantly, but declined 96 h after betamethasone took effect. After 48 h, the percentage of S-phase cells decreased significantly (28.77 to 20.42%; P = 0.014); the percent-age of strongly positive vimentin cells decreased significantly (91 to 51%; P = 0.0092); and the ratio of expression of GFAP protein to the internal control ß-actin increased significantly (0.24 to 0.53; P = 0.1). The level of ERK protein did not change significantly 48 and 96 h after the action of betamethasone, and the pERK/ERK ratios were 0.37 and 0.23, respectively, which were significantly reduced compared with the control group (P = 0.028 and 0.006). Betamethasone has a significant effect on the induction and differentiation of U251 cells, and its mechanism may be related to the inhibition of the abnormal activation of the ERK signal pathway.

Screening of molecular markers linked to dwarf trait in crape myrtle by bulked segregant analysis.

Plant height is one of the most important traits of plant architecture as it modulates both economic and ornamental values. Crape myrtle (Lagerstroemia indica L.) is a popular ornamental woody plant because of its long-lasting mid-summer bloom, rich colors, and diversified plant architecture. These traits also make it an ideal model of woody species for genetic analysis of many ornamental traits. To understand the inheritance of plant height and screen for genes modulating plant height in Lagerstroemia, segregation of the plant height trait was analyzed using the F1 population of L. fauriei (standard) x L. indica 'Pocomoke' (dwarf) with 96 seedlings, while dwarf genes were screened using the bulked segregant analysis method, combined with 28 amplified fragment length polymorphism primers and 41 simple sequence repeat primers. The results showed that the dwarf trait of crape myrtle was controlled by a major gene and modified by minor genes. An amplified fragment length polymorphism marker, M53E39-92, which was 23.33 cM from the loci controlling the dwarf trait, was screened. These results provide basic information for marker-assisted selection in Lagerstromia and cloning of dwarf genes in future studies.

Effects of corn and soybean meal types on rumen fermentation, nitrogen metabolism and productivity in dairy cows.

Twelve multiparous Holstein dairy cows in mid-lactation were selected for a replicated 4×4 Latin square design with a 2 ×2 factorial arrangement to investigate the effects of corn and soybean meal (SBM) types on rumen fermentation, N metabolism and lactation performance in dairy cows. Two types of corn (dry ground [DGC] and steam-flaked corn [SFC]) and two types of SBM (solvent-extracted and heat-treated SBM) with different ruminal degradation rates and extents were used to formulate four diets with the same basal ingredients. Each period lasted for 21 days, including 14 d for adaptation and 7 d for sample collection. Cows receiving SFC had a lower dry matter (DM) and total N intake than those fed DGC. However, the milk yield and milk protein yield were not influenced by the corn type, resulting in higher feed and N utilization efficiency in SFC-fed cows than those receiving DGC. Ruminal acetate concentrations was greater and total volatile fatty acids concentrations tended to be greater for cows receiving DGC relative to cows fed SFC, but milk fat content was not influenced by corn type. The SFC-fed cows had lower ruminal ammonia-N, less urea N in their blood and milk, and lower fecal N excretion than those on DGC. Compared with solvent-extracted SBM-fed cows, cows receiving heat-treated SBM had lower microbial protein yield in the rumen, but similar total tract apparent nutrient digestibility, N metabolism measurements, and productivity. Excessive supply of metabolizable protein in all diets may have caused the lack of difference in lactation performance between SBM types. Results of the present study indicated that increasing the energy degradability in the rumen could improve feed efficiency, and reduce environmental pollution.

Effects of the processing methods of corn grain and soybean meal on milk protein expression profiles in dairy cows.

A proteomic approach was used to investigate the effects of the processing method of corn grain and soybean meal on the milk protein expression profile in lactating dairy cows. A total of 12 multiparous Holstein dairy cows were used in a 4×4 Latin square design with a 2×2 factorial arrangement. The primary factors examined were corn (finely ground (FGC) v. steam-flaked (SFC)) and soybean meal (solvent-extracted (SSBM) v. heat-treated (HSBM)), which were used to formulate four diets with the same basal ingredient: 27% FGC and 9% SSBM; 27% SFC and 9% SSBM; 27% FGC and 9% HSBM; and 27% SFC and 9% HSBM. Each period lasted for 21 days. Milk samples were collected on days 18, 19 and 20 of each period. Changes in the milk proteins were assessed by two-dimensional (2D) electrophoresis and ImageMaster 2D Platinum 6.0 software. A total of 13 spots displayed variations in protein spot abundance according to the statistical analysis. These spots were identified by a matrix-assisted laser desorption/ionization-time of flight/time of flight MS. According to the gels, the relative abundance of α(s2)-casein (CN) fragments was higher in the cows fed the SFC-HSBM than that for SFC-SSBM, whereas ß-CN, α-lactalbumin and zinc-alpha-2-glycoprotein fragments were down-regulated in HSBM-fed cows. The relative decrease of ß-CN expression was validated by western blot and agreed with the MS data. These results suggested that the method used to process soybean meal modified the synthesis and secretion of milk proteins in lactating dairy cows' mammary glands.

Optical layout and mechanical structure of polarimeter-interferometer system for Experimental Advanced Superconducting Tokamak.

A Far-InfaRed (FIR) three-wave POlarimeter-INTerferometer (POINT) system for measurement current density profile and electron density profile is under development for the EAST tokamak. The FIR beams are transmitted from the laser room to the optical tower adjacent to EAST via ∼20 m overmoded dielectric waveguide and then divided into 5 horizontal chords. The optical arrangement was designed using ZEMAX, which provides information on the beam spot size and energy distribution throughout the optical system. ZEMAX calculations used to optimize the optical layout design are combined with the mechanical design from CATIA, providing a 3D visualization of the entire POINT system.

Insertion depth of oral stomach tubes may affect the fermentation parameters of ruminal fluid collected in dairy cows.

Six rumen-fistulated dairy cows were used in 2 trials to validate the technique for the collection of ruminal fluid by an oral stomach tube (OST). Trial 1 was conducted to compare the differences of ruminal fermentation parameters among rumen sites (cranial dorsal, cranial ventral, central, ventral, caudal dorsal, and caudal ventral). The ruminal fluid was collected once per day for 3 consecutive days through rumen cannula (RC). The samples were analyzed for pH, volatile fatty acids (VFA), ammonia N, sodium, potassium, calcium, chloride, and phosphorus concentrations. The ruminal fermentation parameters varied significantly among rumen sites. Compared with the central or ventral rumen, the cranial dorsal rumen had significantly higher pH, ammonia, and sodium concentrations and lower acetate, propionate, and butyrate concentrations, indicating that the sampling site may be one of the main factors contributing to the difference in ruminal fermentation parameters between the samples collected via the OST and RC. In trial 2, the fermentation parameters of ruminal fluid collected via OST at 2 insertion depths (180 or 200 cm) were compared with those of ruminal fluid collected via RC (ventral sac). Ruminal fluid was collected once per week at 5 to 6h after morning feeding. When the OST was inserted to a depth of 180 cm, the OST head was located in the cranial dorsal (atrium) of the rumen. The ruminal fluid collected via the OST had higher pH and sodium concentrations but lower VFA, potassium, calcium, and phosphorus concentrations than that collected via RC. However, when the OST was inserted to a depth of 200 cm, the OST head could pass through the front rumen pillar and reach the central rumen for sampling. No differences were found in pH, VFA, ammonia N, and ion concentrations between the samples collected via the 2 sampling methods. These results indicated that the OST should be inserted to reach the central rumen to obtain representative rumen fluid samples.

Pathway for the elimination of melamine in lactating dairy cows.

Melamine might be degraded into cyanuric acid and some other analogs by the rumen microorganism. Thus, the metabolism of melamine in ruminants may be different from that in monogastric animals. The objective of this study was to investigate the pathway for the elimination of melamine in lactating dairy cows. Four late-lactation dairy cows (body weight=524±17 kg, days in milk=265±14 d) fitted with ruminal cannulas were dosed with melamine (purity ≥99.5%) at 800 mg/d per cow that divided into 2 equal daily doses. The trial lasted for 20 d (13-d preliminary period, followed by a 7-d sample-collecting period). The method of liquid chromatography and tandem mass spectrometry was used to determine melamine and cyanuric acid contents simultaneously. Before the trial started, no melamine or cyanuric acid was detected in samples of total mixed ration, milk, plasma, urine, and feces. The melamine concentration in rumen fluid (Y, mg/L) decreased exponentially after the morning feeding (X, h) (Y=3.85591e(-X/9.25674)+1.35924, R(2)=0.99), but no cyanuric acid was detected. Plasma melamine concentration (0.296±0.014 mg/L) was relatively stable in the 3 different sampling times. The percentages of melamine excreted through milk, urine, and feces were 0.48±0.06, 44.07±10.79 and 10.98±3.88%, respectively. It could be inferred that 44.47±7.98% of ingested melamine was degraded in the rumen, because cyanuric acid was detected in plasma, urine, and feces on the condition that no melamine was contained in the total mixed ration fed to the dairy cows. The results of the present study implied that the elimination pathway of melamine in lactating dairy cows was different from that in monogastric animals. A high percentage of melamine was degraded into cyanuric acid gradually by rumen microorganisms. Most ingested melamine was excreted in urine and feces, which are the main elimination pathways for melamine in lactating dairy cows.

Residues of melamine and cyanuric acid in milk and tissues of dairy cows fed different doses of melamine.

Melamine (1,3,5-triazine-2,4,6-triamine) may be degraded into cyanuric acid and some other compounds by rumen microorganisms. This study was conducted to assess the transfer of melamine and cyanuric acid in to milk and tissues by dairy cows fed different doses of melamine. Forty mid-lactation dairy cows (157±43 d in milk, 20.8±1.4 kg of milk/d) were divided into 4 groups (n=10/group) using a completely randomized design. The groups were fed the following amounts of melamine (purity ≥99.5%) at 0 (control), 300 [treatment (Trt) 1], 500 (Trt 2), and 1,000 (Trt 3) mg/d per cow, respectively. The trial lasted for 18 d (12-d feeding period, followed by a 6-d clearance period). Milk samples were collected from the 4 groups on d 1, 2, 3, 4, 8, 12, 13, 14, 15, and 18, and analyzed for melamine and cyanuric acid. On d 13, 3 cows from Trt 2 and Trt 3 were randomly selected and slaughtered; tissue samples including kidney, liver, mammary, bladder, gluteus medius, and longissimus dorsi were collected for melamine and cyanuric acid analyses. Milk and tissue samples were analyzed for melamine and cyanuric acid using a simultaneous liquid chromatography tandem mass spectrometry procedure. Neither melamine nor cyanuric acid was detected in concentrated feed that was being fed to the cows. In melamine-treated groups, milk melamine concentration increased quickly and reached a stable level by d 4 and was at similar levels on d 8 and 12 after the first administration of melamine. Milk melamine levels in treated groups were 0.18, 0.27, and 0.50mg/L for Trt 1, Trt 2, and Trt 3, respectively, and were highly correlated (R(2)=0.91) with melamine dosing levels. No cyanuric acid was detected in any of the milk collected from the various groups. Melamine residue levels in tissues of Trt 3 were about 2-fold higher than that in Trt 2, with the highest concentration being found in kidney. No differences in cyanuric acid levels in tissues were found between Trt 3 and Trt 2. Liver, kidney, and bladder tissues were found to contain the highest cyanuric acid levels. This study shows a relationship between dietary melamine levels and cyanuric acid levels found in tissues, which might be the result of melamine being converted to cyanuric acid by microorganisms in the rumen.

Transfer efficiency of melamine from feed to milk in lactating dairy cows fed with different doses of melamine.

This study was conducted to investigate the transfer efficiency of melamine (1,3,5-triazine-2,4,6-triamine) from feed to milk of lactating cows fed with different doses of melamine. Twenty-four China Holstein dairy cows were divided into 2 blocks according to milk yield (block 1 and block 2 for low- and high-producing cows). Cows of block 1 or block 2 each were randomly assigned to 1 of 4 treatments in a randomized complete block design and each treatment had 6 cows. The cows of treatments 1 to 4 were dosed with melamine at 0 (control), 90 (treatment 1), 270 (treatment 2), and 450 (treatment 3) mg/d per cow, respectively. The trial lasted 19 d. During the first 13 d, cows were fed melamine at the respective treatment levels, and the last 6 d was the clearance period after melamine was withdrawn. The results indicated that the levels of melamine used did not affect milk yield or composition. The mean milk melamine concentration increased during the initial 3 d after melamine feeding in all the melamine-supplemented groups, and then fluctuated slightly over the remaining 10 d of melamine feeding. No melamine was detected in the milk of any groups on d 4 of the clearance period. Milk melamine concentration measured between 3 to 13 d was significantly affected by melamine feeding doses, but was not influenced by milk yield. The transfer efficiency of melamine from feed to milk was not affected by melamine doses (0.95, 0.70, and 0.66% for treatments 1, 2, and 3, respectively), but was linearly related with milk yield (0.56% for block 1 and 0.95% for block 2, R(2)=0.80). The milk melamine concentration was linearly related with melamine intake (R(2)=0.84). The present study demonstrated that when the daily intake of melamine exceeds 312.7mg/cow, the milk should not be used to produce infant formula powder.

Effects of heating temperatures and addition of reconstituted milk on the heat indicators in milk.

The objective of the present study was to evaluate the effect of heating temperatures and reconstituted milk on heat treatment indicators in milk by comparing the heat damage between raw milk and raw milk plus reconstituted milk (composite milk). The contents of lactulose, furosine, beta-lactoglobulin, and lactoperoxidase were determined after the heat indicators were heated to 65 to 115 °C for 15 s both in raw milk and composite milk. In the raw milk, the lactulose and furosine contents increased with increased heating temperature, while the beta-lactoglobulin content and lactoperoxidase activity decreased. The lactulose and furosine contents were increased after the addition of reconstituted milk. The reconstituted milk also significantly (P < 0.05) reduced the concentration of beta-lactoglobulin in the milk. Both heat treatment and an addition of reconstituted milk decreased the lactoperoxidase activity significantly (P < 0.05), and the lactoperoxidase activity was undetectable at 85 °C. The ratios of lactulose to furosine in pasteurized milk were higher than that in composite pasteurized milk. It is concluded that lactulose, furosine, and beta-lactoglobulin are suitable indicators of high heat pasteurization or raw milk, while lactoperoxidase may be used in monitoring mild heat pasteurization. Practical Application: Adequate heat treatment is necessary to destroy the microbes in raw milk. However, excessive heat treatment can result in inactivation of active compounds or loss of nutrients. The present study showed that the concentrations of lactulose, furosine, beta-lactoglobulin, and the activity of lactoperoxidase are sensitive to processing temperature and can serve as indicators of milk pasteurization.

Antidepressant effect of Yueju-Wan ethanol extract and its fractions in mice models of despair.

AIM OF THE STUDY: Yueju-Wan (YJ), a traditional Chinese medicinal formula, is commonly used for the treatment of depression-related syndromes in China. This study was conducted to evaluate the antidepressant activity of YJ ethanol extract (YJ-E) and its four different fractions, the petroleum ether fraction (YJ-EA), ethyl acetate fraction (YJ-EB), n-butanol fraction (YJ-EC) and final aqueous fraction (YJ-ED). MATERIALS AND METHODS: Two experimental despair animal models: the mice tail suspension test (TST) and the mice forced swimming test (FST) were used to evaluate the antidepressant activity of YJ-E and its fractions. These extracts or fractions were administered orally for 7 days, while the parallel positive control was given at the same time using fluoxetine hydrochloride (FLU) in TST and imipramine hydrochloride (IMI) in FST respectively. RESULTS: YJ-E high dose (YJ-E2), YJ-EA, YJ-EC and the positive control groups could decrease the duration of immobility in the TST and FST and have no significant changes in locomotor activity. YJ-E low dose (YJ-E1), YJ-EB, YJ-ED and the vehicle solvent (VEH) control group have no obvious effect on these same tests. CONCLUSIONS: In these despair animal models, YJ ethanol extract, the petroleum ether fraction and n-butanol fraction show potent antidepressant effects. The petroleum ether fraction and n-butanol fraction appear to be the active fractions of YJ-E.

Brain transplantation of genetically modified bone marrow stromal cells corrects CNS pathology and cognitive function in MPS VII mice.

Current therapies for lysosomal storage diseases (LSDs), enzyme replacement therapy and bone marrow transplantation are effective for visceral organ pathology of LSD, but their effectiveness for brain involvement in LSDs is still a subject of controversy. As an alternative approach, we transplanted genetically modified bone marrow stromal (BMS) cells to lateral ventricle of newborn mucopolysaccharidosis VII (MPS VII) mice. MPS VII is one of LSDs and caused by deficiency of beta-glucuronidase (GUSB), resulting in accumulation of glycosaminoglycans (GAGs) in brain. At 2 weeks after transplantation, the GUSB enzyme-positive cells were identified in olfactory bulb, striatum and cerebral cortex, and the enzymatic activities in various brain areas increased. The GAGs contents in brain were reduced to near normal level at 4 weeks after transplantation. Although GUSB activity declined to homozygous level after 8 weeks, the reduction of GAGs persisted for 16 weeks. Microscopic examination indicated that the lysosomal distention was not found in treated animal brain. Cognitive function in MPS VII animals as evaluated by Morris Water Maze test in treated mice showed a marked improvement over nontreated animals. Brain transplantation of genetically modified BMS cells appears to be a promising approach to treat diffuse CNS involvement of LSDs.

Treatment of lysosomal storage disorders: cell therapy and gene therapy.

Most lysosomal storage diseases have central nervous system (CNS) involvement. No effective treatment is available at present. We investigated the usefulness of brain-directed gene therapy and cell therapy using mouse models of lysosomal storage diseases. For gene therapy to the CNS, a recombinant adenovirus encoding beta-galactocerebrosidase gene was injected into the cerebral ventricle of neonatal twitcher mice, a murine model of Krabbe disease. Improvements in neurological symptoms and a prolonged lifespan were observed. Brain activity of beta-galactocerebrosidase was increased significantly and the concentration of a cytotoxic metabolite, psychosine, was decreased. Pathological observations of the brain were also improved in treated twitcher mice. For cell therapy to the CNS, a neural stem cell line derived from human fetal brain was genetically engineered to overexpress beta-glucuronidase and transplanted into the cerebral ventricles of neonatal MPS VII mice, a model of beta-glucuronidase deficiency. Transplanted human neural stem cells were found to integrate and migrate in the host brain and to produce large amounts of beta-glucuronidase. Brain contents of the substrate of beta-glucuronidase were reduced and widespread clearing of lysosomal storage was observed in treated MPS VII mice. These data suggest that brain-directed gene/cell therapy may be useful in the treatment of neurological alterations in lysosomal storage diseases.

Adenovirus-mediated prenatal gene transfer to murine central nervous system.

In some lysosomal storage disorders pathological alterations in the central nervous system (CNS) occur as early as the prenatal period and the neuropathology progresses rapidly soon after birth. In these diseases, postnatal therapies alone are often insufficient. Therefore prenatal gene therapy to the CNS may be necessary. In order to investigate the feasibility of gene transfer to the CNS prenatally, we administered recombinant adenovirus carrying LacZ gene to rat embryos from embryonic day 9 to 12 (E9-E12). Results showed that efficient transduction of the reporter gene to the CNS was achieved when adenoviruses were injected at E12. The regions where the reporter gene was transduced mainly localized at the telencephalon and hypophysis of the embryo, and the gene expression persisted at least 1 week after birth. In addition, when adenoviruses were injected at E9, E10 and E11, no transgene expression was detected in the CNS, but was mainly observed in the liver, the heart and the skin, respectively.

Intraventricular administration of recombinant adenovirus to neonatal twitcher mouse leads to clinicopathological improvements.

Twitcher mouse is a murine model of human globoid cell leukodystrophy (Krabbe disease), which is characterized by a genetic deficiency in galactocerebrosidase (GALC) activity. The nervous system is affected early and severely by demyelination in the white matter. So far, there is no effective treatment for Krabbe disease except bone marrow transplantation (BMT). However, BMT has inherent limitations such as unavailability of donors and graft-versus-host disease. In this study, we injected recombinant adenovirus encoding GALC into the lateral ventricle of twitcher mice at postnatal day 0 (PND 0) and the therapeutic effects were evaluated. Our results showed slight, but significant improvements in motor functions, body weight and twitching and a prolonged life span. In brain, GALC activity was increased to 15% that of normal littermates and psychosine concentration was decreased to 55% that of untreated twitcher mice at PND 15. The number of PAS-positive globoid cells in brain stem was also reduced significantly at PND 35. In contrast, when adenoviruses were injected to the twitcher mice at PND 15, almost no improvements were observed. These results demonstrate that the timing of treatment may be of great importance in Krabbe disease.

Genetically modified bone marrow continuously supplies anti-inflammatory cells and suppresses renal injury in mouse Goodpasture syndrome.

In chronic inflammation, macrophages and neutrophils, which are derived from bone marrow, play a pivotal role. Therefore, reconstitution of bone marrow with anti-inflammatory stem cells may modify inflammation. In this study, transplantation-based gene therapy was applied to glomerular inflammation for a long-lasting suppression of the glomerular damage seen in chronic nephritis. Bone marrow cells were harvested from male donor mice, which had received 5-fluorouracil 3 days previously, and transduced with an interleukin 1 (IL-1) receptor antagonist (IL-1Ra) or a mock gene using a retrovirus vector. After confirmation that transduced cells possessed the transgene at approximately 0.7 copies per cell and secreted recombinant IL-1Ra, these cells were infused into sublethally irradiated (6 Gy) female recipients once daily for 4 consecutive days. These female recipient mice had the male Y antigen in bone marrow, liver, and spleen, and 10% to 20% of their spleen cells possessed the transgene even 8 weeks after transplantation. Glomerulonephritis was then induced in these mice. Renal function and histology were retarded in the mice whose bone marrow was reconstituted with IL-1Ra-producing cells compared with mock transduced cells. In situ hybridization using a Y painting probe revealed that transplanted donor cells were recruited into the glomerulus upon induction of nephritis, suggesting therapeutic effects were channeled through the secretion of IL-1Ra from these cells. Furthermore, the survival rate after a second challenge with nephrotoxic antibody was significantly improved in the IL-1Ra chimera. These results suggest that reconstitution of bone marrow for continuous supply of anti-inflammatory cells may be a useful strategy for the treatment of chronic inflammation.