Changes in the Metabolome of Picea balfouriana Embryogenic Tissues That Were Linked to Different Levels of 6-BAP by Gas Chromatography-Mass Spectrometry Approach.

Embryogenic cultures of Picea balfouriana, which is an important commercial species for reforestation in Southern China, easily lose their embryogenic ability during long-term culture. Embryogenic tissue that proliferated at lower concentrations (3.6 µM and 2.5 µM) of 6-benzylaminopurine (6-BAP) were more productive, and generated 113 ± 6 and 89 ± 3 mature embryos per 100 mg embryogenic tissue, respectively. A metabolomic approach was used to study the changes in metabolites linked to embryogenic competence related to three different 6-BAP concentrations (2.5 µM, 3.6 µM, and 5 µM). A total of 309 compounds were obtained, among which 123 metabolites mapped to Kyoto Encyclopedia of Genes and genomes (KEGG) pathways. The levels of 35 metabolites were significantly differentially regulated among the three 6-BAP treatments, and 32 metabolites differed between the 2.5 µM and 5 µM treatments. A total of 17 metabolites appeared only once among the three comparisons. The combination of a score plot and a loading plot showed that in the samples with higher embryogenic ability (3.6 µM and 2.5 µM), up-regulated metabolites were mostly amino acids and down-regulated metabolites were mostly primary carbohydrates (especially sugars). These results suggested that 6-BAP may influence embryogenic competence by nitrogen metabolism, which could cause an increase in amino acid levels and higher amounts of aspartate, isoleucine, and leucine in tissues with higher embryogenic ability. Furthermore, we speculated that 6-BAP may affect the amount of tryptophan in tissues, which would change the indole-3-acetic acid levels and influence the embryogenic ability.

Human leucocyte antigen-Bw4 and Gag-specific T cell responses are associated with slow disease progression in HIV-1B-infected anti-retroviral therapy-naive Chinese.

In China, the majority of human immunodeficiency virus (HIV) infections are predominately subtype B. It is important to characterize the HIV-1 subtype B-specific and its T cell response within the Chinese population, with the aim of identifying protective correlates of immunity to control HIV-1 infections. In this study, we performed a comprehensive analysis looking into the magnitude/strength of T cell responses directed at the Gag protein of the HIV-1 subtype B, one of the most conserved HIV-1 proteins. The study group consisted of anti-retroviral native and chronic HIV-1 subtype B-infected individuals. We used enzyme-linked immunospot (ELISPOT) assay to quantify the total T cell responses to HIV-1 Gag at the single peptide level. Twenty-eight (38%) peptides were recognized in 24 (82·8%) individuals. The p24 was identified as the most frequently recognized subunit protein with the greatest T cell response in the test, which correlated positively with CD4(+) T cell count and inversely with viral load (VL). At the level of the human leucocyte antigen (HLA) supertypes, we detected the highest levels and a significant correlation with both the CD4(+) T cell count and the VL with Gag T cell responses in Bw4/Bw4. These findings demonstrate that (i) the HIV-1B Gag p24-specific immune responses play an important role in controlling viral replication and slowing clinical progression; and (ii) HLA-Bw4/Bw4 allele has stronger T cell responses, which is associated with slow clinical progression in Chinese HIV patients.

Magnetic resonance imaging in determination of myocardial ischemia and viability: comparison with positron emission tomography and single-photon emission computed tomography in a porcine model.

BACKGROUND: In patients with chronic left ventricular dysfunction, the size of the viable cardiac muscle is correlated with the prognosis and the outcome of myocardial revascularization. PURPOSE: To evaluate the diagnostic value of various imaging techniques in determination of myocardial ischemia and viability. MATERIAL AND METHODS: A chronic myocardial ischemia animal model was established, in which 10 pigs underwent magnetic resonance imaging (MRI), positron emission tomography (PET), and single-photon emission computed tomography (201Tl SPECT) before and 1-2 months after modeling. The size of myocardial ischemia and necrosis was judged, and the imaging manifestations were compared with pathologic findings. RESULTS: Seven of the 10 animals completed all examinations uneventfully. On dobutamine-stressed cine MRI, 10 (8.93%) segments were found to be akinetic. Perfusion was abnormal in 34 (30.35%) segments. Delayed hyperenhancement was observed in 12 (10.71%) segments. PET detected myocardial necrosis in 17 (15.18%) segments, and SPECT detected myocardial necrosis in nine (8.04%) segments. Histological examination with triphenyltetrazolium chloride (TTC) showed pale necrosis in 14 (12.50%) segments. The number of necrotic segments detected by PET was significantly greater than that by contrast-enhanced MRI (chi2 = 5, P = 0.0253, kappa = 0.8028) and cine MRI (chi2 = 7, P = 0.0082, kappa = 0.7079). It was also greater than that by TTC (chi2 = 3, P = 0.0833, kappa = 0.8879), although the difference was statistically insignificant. The number of necrotic segments detected by SPECT was significantly smaller than that by TTC (chi2 = 5, P = 0.0253, kappa = 0.7590), as was the number of necrotic segments detected by cine MRI (chi2 = 4, P = 0.0455, kappa = 0.8100). There was no statistically significant difference in the detection of necrotic segments between contrast-enhanced MRI and TTC (chi2 = 2, P = 0.1573, kappa = 0.9130). CONCLUSION: Cardiac MRI can determine viable myocardium and clearly delineate the location and degree of myocardial necrosis. PET slightly overestimates the extent of the necrotic myocardium and is unable to distinguish transmural necrosis from subendocardial necrosis.

A quantitative (stereological) study of the effects of experimental unilateral cryptorchidism and subsequent orchiopexy on spermatogenesis in adult rabbit testis.

The aim of this study was to examine the controversial effects of experimental unilateral cryptorchidism and subsequent orchiopexy on the number of germ cells and other morphometric characteristics of testicular and epididymal structures in adult rabbits. Unilateral cryptorchidism was induced in 11 mature male New Zealand white rabbits by returning one testis, together with the ipsilateral epididymis, to the abdominal cavity via a surgical procedure. After 3 months, testes and epididymides were removed from six animals (and from six age-matched control animals that did not undergo the surgery). Orchiopexy was performed on the five remaining animals and the testes and epididymides of these animals (and an additional six age-matched control animals) were removed 7 weeks later. A contemporary, unbiased and efficient stereological tool, the optical disector, was used to estimate the number of nuclei in the testis and epididymis using methacrylate-embedded sections of 25 micron in thickness. Cryptorchidism resulted in severe testicular atrophy and spermatogenic arrest: type A spermatogonia and Sertoli cells only were seen in the seminiferous epithelium, and the number of type A spermatogonia per testis was reduced by 84%. After orchiopexy, the testis remained atrophied and the number of type A spermatogonia returned to the near-normal range in four of five animals, but spermatogenesis was recovered only partially at the stage of early primary spermatocytes (one animal), late primary spermatocytes (two animals) or spermatids (one animal). In conclusion, cryptorchidism caused severe spermatogenic arrest that was potentially recoverable (in view of the restoration of the number of type A spermatogonia), but orchiopexy failed to induce full recovery of spermatogenesis.

Idiopathic sudden hearing loss and disturbance of iron metabolism. A clinical survey of 426 cases.

The role of disturbances of the iron metabolism (DIM) in idiopathic sudden hearing loss (ISHL) was investigated in 426 patients with ISHL who received iron therapy, anti-DIM medication, vitamins, and a combined regimen in a randomized study which was stratified by stages and groups. Low concentrations of haemoglobin, serum iron, serum ferritin, and red cell basic ferritin as well as abnormal circadian variations in the serum iron level were observed. The results were significantly better in patients receiving iron therapy than in those receiving anti-DIM medication, vitamins, and the combined regimen. Hearing improvement was achieved in 53.26% of the patients whose treatment started later than 3 months after the onset of the disease. The clinical association of DIM and ISHL is discussed.

Neuroendocrine evaluation in Kleine-Levin syndrome: evidence of reduced dopaminergic tone during periods of hypersomnolence.

A patient with Kleine-Levin syndrome had polysomnography and neuroendocrinological assays performed during asymptomatic (ASMP) and symptomatic (SMP) 24-hr periods. During the SMP, sleep data revealed poor nocturnal sleep efficiency, increased sleep fragmentation and reduced stages 3, 4 and rapid eye movement (REM). No sleep onset REM episodes were seen. Sleep staging in the ASMP was normal. Blood samples were obtained every 20 min and assayed for thyroid-stimulating hormone (TSH), cortisol (CORT), prolactin (PRL) and growth hormone (GH). Patterns of secretion, 24-hr mean and total integrated concentrations, and mean sleep period time values during the ASMP and SMP were compared. The mean 24-hr level of TSH was increased and GH decreased in the SMP. Comparing sleep period time in the SMP to the ASMP, values for TSH and PRL were increased and GH and CORT were reduced in the SMP. These hormone changes support the hypothesis that reduced hypothalamic dopaminergic tone is present in the SMP compared to the ASMP in Kleine-Levin patients.

Red cell basic ferritin concentration in sensorineural hearing loss.

Red cell basic ferritin (RCBF) was measured in 62 healthy subjects with normal hearing and 224 patients with different kinds of sensorineural hearing loss, compared with serum ferritin, haemoglobin, serum iron and circadian variation in the serum iron levels. The results showed that the RCBF concentrations in most kinds of sensorineural hearing loss were significantly lower than those in normal hearing status. It is conceivable that the RCBF assay can be used to evaluate the adequacy of iron stores that are useful in times of urgent iron needs and to diagnose relative iron deficiency in those patients with some other normal laboratory assays related to iron metabolism.

Dynamic observation on thyroid function in severe head injury.

The dynamic changes of thyroid hormone levels in patients with severe head injury were observed. The results showed that the serum thyroid hormone levels changed rapidly within several hours after the injury and the mean levels of serum total T3 and T4 (TT3, TT4) decreased while high reverse triiodothyronine (rT3) increased significantly. Serum TT3, TT4 and rT3 levels returned to normal gradually in 2 to 3 weeks after the injury in the surviving group, whereas serum total T4 gradually declined and rT3 steadily increased in the fatal group. The mean levels of serum TSH remained unchanged in both groups. Therefore, the use of the levels of serum thyroid hormones as adjuvant indices can be helpful in making pertinent judgements of the severity and prognosis of severe head injuries.