YTHDC2-mediated m6A mRNA modification of Id3 suppresses cisplatin resistance in non-small cell lung cancer.

Background: The methylation of adenosines at the N6 position (N6-methyladenosine; m6A) is one of the most conserved internal RNA modifications. m6A can modulate the expression of oncogenes or tumor suppressor genes, as well as m6A levels and the expression and activity of m6A enzymes, thus influencing tumor progression and therapeutic response. This study investigates the role of YTHDC2-mediated m6A messenger RNA (mRNA) modification of Id3 in controlling cisplatin resistance in non-small cell lung cancer (NSCLC). Methods: The expression of the m6A reader protein YTHDC2 was detected in an NSCLC cisplatin-resistant cell line (A549/DDP) using real-time fluorescence quantitative polymerase chain reaction (qPCR). YTHDC2 overexpression plasmids were constructed and transfected into A549/DDP and A549 cells respectively. We performed qPCR and western blot (WB) to detect changes in YTHDC2 and Id3 expression, and the effects of YTHDC2 overexpression on proliferation, apoptosis, invasion, and migration of drug-resistant cells were assessed by cell counting kit-8 (CCK-8), flow cytometry, and transwell and scratch assays. The m6A modification of Id3 by YTHDC2 was clarified by m6A-immunoprecipitation-PCR (m6A-IP-PCR) assay. Results: The CLIPdb online database predicted that YTHDC2 might bind to Id3. The results of qPCR showed that YTHDC2 was downregulated in the NSCLC cisplatin-resistant cell line A549/DDP compared to the cisplatin-sensitive cell line A549. Overexpression of YTHDC2 increased the expression of Id3, and the methylation inhibitor 3-deazaadenosine abrogated the regulatory effect of YTHDC2 on Id3. YTHDC2 overexpression significantly inhibited A549/DDP cell proliferation, migration, and invasion, and promoted apoptosis by synergistically promoting the effects of Id3. m6A-IP-PCR analysis revealed that YTHDC2 could inhibit the m6A level of Id3 mRNA. Conclusions: To regulate the activity of Id3, YTHDC2 requires modifications to m6A, which ultimately inhibit cisplatin resistance in NSCLC.

Decreased expression of HBV surface antigen (HBsAg) with sK122R and sV96A co-mutation is associated with an ineffective antibody response in a chronic hepatitis B patient.

BACKGROUND: Emergence and predominance of hepatitis B virus (HBV) variants carrying S gene mutations frequently occur in HBV-infected individuals. Here, coexistent serum anti-HBsAg antibody (HBsAb) and HBV surface antigen (HBsAg) were detected in a chronic HBV patient. The patient's HBsAg proteins possessed amino acid substitutions sK122R and sV96A. We reported this case and conducted relevant studies to investigate differences in expression levels and antibody neutralization of HBsAg proteins bearing sK122R and sV96A amino acid substitutions to explore causes of antigen-antibody coexistence in a chronic hepatitis B patient. STUDY DESIGN: We first sequenced the S gene from HBV present within the patient's serum. Based on the S gene sequence, we cloned wild-type and mutated S gene sequences via site-directed mutagenesis to construct expression plasmids pJW4303-WT (wild-type), pJW4303-sV96A, pJW4303-sK122R, and pJW4303-sV96A-sK122R. Plasmids were transfected into HEK 293 T cells then culture supernatants and cells were collected. Collected cells and supernatants were next subjected to a series of quantitative and functional tests to assess expression and neutralization characteristics of wild-type and mutant HBsAg proteins. RESULTS: Based on quantification of HBsAg expression in cells transfected with the four plasmids, HBsAg-sK122R-sV96A was more intracellularly retained and less secreted than HBsAg-sV96A single-mutant protein and WT. Neutralization ability of serum from chronic HBV patient against culture supernatants containing recombinant HBsAg proteins were ranked from highest to lowest as HBsAg-sV96A, HBsAg-sV96A-sK122R, and HBsAg-sK122R. However, no significant differences of neutralization efficiency by high-potency antibodies from HBV-vaccinees against these three mutant proteins were observed. CONCLUSIONS: The levels of HBsAg proteins with amino acid substitutions sV96A-sK122R were greatly reduced in culture supernatants but were apparently increased in the intracellular fraction. This may account for the higher levels of HBV replication in patients. HBsAg neutralization by HBsAb in this patient may have been compromised by the HBsAg sK122R amino acid substitution, suggesting that antibodies produced by the patient had lost their HBV-neutralizing effect.

Measurement of echo reduction for passive-material samples using sparse Bayesian learning and least squares estimation.

Echo reduction (ER) for passive materials is important for the evaluation of sound absorption performance. In a limited space, due to the strong interference of multipath signals, it is difficult to separate and extract the directed and reflected signals of the sample from the measured signal to accurately calculate the ER, especially at low frequencies. A method combining sparse Bayesian learning (SBL) and the least squares estimation (LSE) is proposed to extract the directed and normal reflected signals of the sample from the received signal. First, owing to the high resolution of SBL in time delays estimation, the set of multipath time delays is estimated. Then, the LSE is utilized to evaluate the amplitudes of multipath signals with estimated time delays as a priori information. With combination processing, the resolution of time delay estimation is enhanced, the dimension of the LSE is reduced, and the accuracy of the amplitude estimation for the directed and normal reflected signals, as well as the ER evaluation, is improved. The proposed method is validated through simulations and experiments in a cylindrical tank.

Expression profiles of circular RNAs in colon biopsies from Crohn's disease patients by microarray analysis.

BACKGROUND: Circular RNAs (circRNAs) are involved in various diseases and serve as biomarkers. The present study aimed to investigate unique expression profiles of circRNAs in colon tissues of Crohn's disease (CD) and search novel biomarkers for the diagnosis. METHODS: Differentially expressed (DE) circRNAs in biopsies from four CD patients, four ulcerative colitis (UC) patients, and four healthy controls (HC) were screened by microarray. Hsa_circ_0062142 and hsa_circ_0001666 were verified in another expanded validation cohort. Bioinformatics analysis was applied to predict the function of two DE circRNAs. Receiver operating characteristic (ROC) curves were constructed to evaluate the diagnostic value of CD. RESULTS: The top 10 upregulated circRNAs in CD compared with HC were hsa_circ_0000691, hsa_circ_0001666, hsa_circ_0004183, hsa_circ_0009024, hsa_circ RNA_405324, hsa_circ_0002003, hsa_circ_0085323, hsa_circ_0040994, hsa_circ_0062142, and hsa_circ_0048148; the top 10 downregulated circRNAs were hsa_circ_0049356, hsa_circ RNA_405443, hsa_circ RNA_403556, hsa_circ_0092328, hsa_circ_0003979, hsa_circ_0074491, hsa_circ_0023461, hsa_circ RNA_406237, hsa_circ_0034044, and hsa_circ RNA_400564 (fold change in descending order). The expression levels of hsa_circ_0001666 and hsa_circ_0062142 in CD were significantly higher than those in UC and HC (p < 0.01). ROC curves suggested the favorable diagnostic value of hsa_circ_0062142 and hsa_circ_0001666 (AUC = 0.803 and 0.858, respectively, p < 0.01). In silico analysis indicated that these circRNAs may be involved in the progress of CD. CONCLUSION: Hsa_circ_0062142 and hsa_circ_0001666 may play critical roles in the pathogenesis and serve as potential biomarkers of CD.

Characterizing Mat Formation of Bamboo Fiber Composites: Horizontal Density Distribution.

Bamboo fiber composite (BFC) is a unidirectional and continuous bamboo fiber composite manufactured by consolidation and gluing of flattened, partially separated bamboo culm strips into thick and dense panels. The composite mechanical properties are primarily influenced by panel density, its variation and uniformity. This paper characterized the horizontal density distribution (HDD) within BFC panels and its controlling factors. It revealed that HDD follows a normal distribution, with its standard deviation (SD) strongly affected by sampling specimen size, panel thickness and panel locations. SD was lowest in the thickest (40 mm) panel and largest-size (150 × 150-mm2) specimens. There was also a systematic variation along the length of the BFC due to the tapering effect of bamboo culm thickness. Density was higher along panel edges due to restraint from the mold edges during hot pressing. The manual BFC mat forming process is presented and found to effectively minimize the density variation compared to machine-formed wood composites. This study provides a basic understanding of and a quality control guide to the formation uniformity of BFC products.

Prenatal diagnosis and genetic counseling in a fetus associated with risk of Angelman syndrome with a small supernumerary marker chromosome derived from chromosome 22.

BACKGROUND: Angelman syndrome (AS) is a neurodevelopmental disorder. AS patients concomitant with sSMC are rather rare events. It will provide more useful and proper information for genetic counseling to identify the sSMC origin. CASE PRESENTATION: A 27-year-old woman was referred for genetic counseling and prenatal diagnosis at 26 weeks of gestation due to her elder daughter, diagnosed as Angelman syndrome (AS) with an interstitial deletion in one of the chromosomes 15, carrying a small supernumerary marker chromosome (sSMC). The G-banding results of the woman and her current fetus both were 47,XX,+mar. In this paper, fluorescence in situ hybridization (FISH) results showed that there was no deletion of chromosome 15 in the woman and fetus. We demonstrated that the proband's sSMC was maternally inherited and was an inv dup(22)(q11.1) , and that the deletion in 15q11.2-q13.1 was de novo. CONCLUSIONS: Taking into account above results and normal phenotypes of the proband's mother, in this case we suggest that the sSMC don't increase the recurrence risk of AS. After prenatal diagnosis, the woman chose to continue the pregnancy, and finally gave birth to a normal female infant.

Distribution of yeast isolates from invasive infections and their in vitro susceptibility to antifungal agents: evidence from 299 cases in a 3-year (2010 to 2012) surveillance study.

Invasive yeast infections cause significant morbidity and mortality. Surveillance for the infection is necessary to detect trends in species distribution and antifungal resistance. We performed this retrospective study of yeast infection at Jinling Hospital, Nanjing in China, from year of 2010 to 2012. A total of 341 yeast isolates were obtained from patients with invasive infections in the period. Among these isolates, Candida spp. comprised of the highest percentage of yeast strains (91.8 %), followed by Cryptococcus neoformans (5.9 %) and other non-Candida yeast strains (2.3 %). Bloodstream isolates made up 41.3 % of yeast strains and the isolates from CVC made up 17.3 %. Among Candida spp., C. albicans was the most common species identified from non-blood clinical specimens (42.9 %), but appeared in only 20.8 % of blood isolates (P < 0.001). C. tropicalis was the most prevalent Candida species in the blood samples (28.5 %). Candida spp. was mainly isolated from specimens of the ICU patients, while C. neoformans was mainly isolated from specimens in medical wards. Resistance to FLC occurred in 3.7 % of C. albicans, 9.9 % of C. tropicalis, 74.0 % of C. glabrata, and 4.4 % of C. parapsilosis. Most (>92 %) isolates of C. albicans, C. tropicalis, C. parapsilosis, and C. neoformans strains were susceptible to VRC; However, 26.7 % of isolates of C. glabrata were VRC resistant.

[Lipopolysaccharide affects testicular histology and reproductive endocrine function in male rats].

OBJECTIVE: To study the influence of lipopolysaccharide (LPS)-induced inflammation on the testicular histology and reproductive endocrine function in male rats and investigate the possible mechanism of inflammation affecting male fertility. METHODS: Thirty-six male SD rats were randomly divided into a control group (A) and three LPS intervention groups (B, C, and D) to receive saline and LPS (5 mg/kg i. p, once), respectively. The animals in groups B, C, and D were killed by anesthesia at 12, 24, and 72 hours after treatment. Histopathological changes in the left testis of the rats were observed by HE staining and the levels of the reproductive hormones T, FSH, and LH in the serum were determined by ELISA. RESULTS: Compared with group B, group A showed clear structure of seminiferous tubules, orderly arrangement of spermatogenic cells, a slightly decreased number of sperm in some seminiferous tubular lumens, and shed spermatogenic cells in the rat testis tissue; group C exhibited thinner seminiferous epithelia, disordered structure of seminiferous tubules, irregular arrangement of spermatogenic cells, decreased number of mature sperm and obvious shedding of spermatogenic cells in seminiferous tubular lumens; group D manifested similar findings to those of group C, with even more shed spermatogenic cells that blocked the tubular lumens. The levels of serum T, LH, and FSH were (0.490 +/- 0.028) ng/ml, (6.290 +/- 0.515) ng/L, and (1.837 +/- 0.127) IU/L in group A, (0.460 +/- 0.024) ng/ml, (5.881 +/- 0.124) ng/L, and (1.707 +/- 0.098) IU/L in group B, (0.417 +/- 0.021) ng/ml, (5.123 +/- 0.271) ng/L, and (1.620 +/- 0.115) IU/L in group C, and (0.378 +/- 0.021) ng/ml, (4.504 +/- 0.279) ng/L and (1.562 +/- 0.216) IU/L in group D, all decreased in group B as compared with A (P > 0.05). The decreases of T and LH were extremely significant (P < 0.01) and that of FSH was significant in groups C and D (P < 0.05) in comparison with A. CONCLUSION: LPS-induced inflammation affects the testicular tissue and reproductive endocrine function of male rats, resulting in decreased levels of serum T, LH, and FSH.

Five-day, low-level laser therapy for sports-related lower extremity periostitis in adult men: a randomized, controlled trial.

Periostitis in the lower leg caused by overexercise is a universal problem in athletes and runners. The purpose of this study was to observe the functional improvement of the lower limbs upon rehabilitation low-level laser therapy (LLLT). All medical data were gathered from enrolled adults with sports-related lower leg pain. A total of 54 patients underwent triple-phase bone scans using skeletal nuclear scintigraphy, which confirmed periostitis in their lower limbs. The patients were then randomly divided into two groups: one group received laser therapy (N = 29) and the other group (N = 25) received an equivalent placebo treatment (a drug or physical therapy). Treatment protocol commenced with rehabilitation intervention and LLLT was performed three times daily for 5 days at a dosage of 1.4 J/cm(2). A Likert-type pain scale was used to evaluate the severity of pain. Balance function, including postural stability testing (PST) and limits of stability (LOS), was also performed to evaluate the function outcome. Patients experienced a significant improvement in pain by day 2 or day 5 after starting LLLT, but here was no significant difference in pain scale between the measurements before (baseline) and after LLLT. Comparing the PST, the group differences of dynamic vs. static testings ranged from -18.54 to -50.22 (compared 12, 8, 4, 3, 2, 1 to 0, all p < 0.0001), and the PST after LLLT were 3.73 units (p = 0.0258) lower than those of before LLLT. Comparing the LOS, the group differences of dynamic vs. static testing were similar to those in PST, and the relationship between LOS and groups only varied with the direction control during dynamic testing in direction at backward/right vs. right (p < 0.0001). LLLT had a positive effect on proprioception in patients with lower limb periostitis. Larger, better controlled studies are needed to determine what specific effects LLLT has on the function of proprioception.

Novel mutations of ABCB6 associated with autosomal dominant dyschromatosis universalis hereditaria.

OBJECTIVE: Dyschromatosis universalis hereditaria (DUH) is a rare heterogeneous pigmentary genodermatosis, which was first described in 1933. The genetic cause has recently been discovered by the discovery of mutations in ABCB6. Here we investigated a Chinese family with typical features of autosomal dominant DUH and 3 unrelated patients with sporadic DUH. METHODS: Skin tissues were obtained from the proband, of this family and the 3 sporadic patients. Histopathological examination and immunohistochemical analysis of ABCB6 were performed. Peripheral blood DNA samples were obtained from 21 affected, 14 unaffected, 11 spouses in the family and the 3 sporadic patients. A genome-wide linkage scan for the family was carried out to localize the causative gene. Exome sequencing was performed from 3 affected and 1 unaffected in the family. Sanger sequencing of ABCB6 was further used to identify the causative gene for all samples obtained from available family members, the 3 sporadic patients and a panel of 455 ethnically-matched normal Chinese individuals. RESULTS: Histopathological analysis showed melanocytes in normal control's skin tissue and the hyperpigmented area contained more melanized, mature melanosomes than those within the hypopigmented areas. Empty immature melanosomes were found in the hypopigmented melanocytes. Parametric multipoint linkage analysis produced a HLOD score of 4.68, with markers on chromosome 2q35-q37.2. A missense mutation (c.1663 C>A, p.Gln555Lys) in ABCB6 was identified in this family by exome and Sanger sequencing. The mutation perfectly cosegregated with the skin phenotype. An additional mutation (g.776 delC, c.459 delC) in ABCB6 was found in an unrelated sporadic patient. No mutation in ABCB6 was discovered in the other two sporadic patients. Neither of the two mutations was present in the 455 controls. Melanocytes showed positive immunoreactivity to ABCB6. CONCLUSION: Our data add new variants to the repertoire of ABCB6 mutations with DUH.

Surveillance study of species distribution, antifungal susceptibility and mortality of nosocomial candidemia in a tertiary care hospital in China.

BACKGROUND: Bloodstream infections due to Candida species cause significant morbidity and mortality, and the epidemiology of Candida infection is changing. Surveillance for candidemia is necessary to detect trends in species distribution and antifungal resistance. METHODS: The medical and electronic records of all patients who had candidemia at the authors' hospital from 2009 to 2011 were reviewed for demographic data and clinical information, including the infecting Candida species, resistance to antifungals and survival, and the presence of risk factors associated with candidemia. RESULTS: A total of 133 distinct episodes of candidemia were identified over the study period. The annual incidence of candidemia ranged between 0.71 and 0.85 cases/1000 hospital discharges. The most frequent Candida species were C. tropicalis (28.6%), followed by C. albicans (23.3%) and C. parapsilosis (19.5%). The rates of susceptibility to antifungal agents were as followed: voriconazole (97.8%), itraconazole (69.5%), fluconazole (46.1%), ketoconazole (38.9%). Out of 131 evaluable patients, 34 (26.0%) died within 30 days from the onset of candidemia. C. tropicalis candidemia was associated with the highest mortality rate (44.7%). Regarding the crude mortality in the different units, patients in Hemato-Oncology ward had the highest mortality rate (66.7%), followed by patients in cardiovascular wards and ICU (57.1% and 25.6%, respectively). Predictors of 30-day mortality were identified by uni- and multivariate analyses. Complicated abdominal surgery, presence of central venous catheter (CVC), neutropenia, candidemia due to C. tropicalis and poor treatment with fluconazole were significantly associated with the 30-day mortality. Presence of CVC (odds ratio[OR] = 4.177; 95% confidence interval [CI] = 1.698 to 10.278; P = 0.002) was the only independent predictor for mortality in the multivariate analysis. CONCLUSION: This report provides baseline data for future epidemiological and susceptibility studies and for the mortality rates associated with candidemia in our hospital. The knowledge of the local epidemiological trends in Candida species isolated in blood cultures is important to guide therapeutic choices.

[Meiotic analysis of spermatogenic cells in severe oligoasthenoteratozoospermia with chromosome 13 rearrangement].

OBJECTIVE: To explore the possible mechanisms of spermatogenic arrest in severe oligoasthenoteratozoospermia induced by supernumerary, ring-neocentric 13q12.3 --> 13q22 chromosome and reciprocal deletion. METHODS: We performed a genomic-wide high-density oaCGH analysis for a case of oligoasthenoteratozoospermia with abnormal chromosome 13 to characterize the breakpoints of the chromosome involved or the gene deletion caused by the rearrangement. We also conducted a fluorescence in situ hybridization analysis on the germ cells using probes of 13q14/13qter to observe the pairing condition of homologous chromosome 13. RESULTS: We identified by oaCGH analysis a microdeletion of 4 consecutive probes (A_16_P19757882, A_16_P02744617, A_14_ P108858 and A_16_P02744687 at chr13q12.3: 27979261 --> 28039191) with 59.93 kb between the FLT1 and POMP genes, with no annotated genes in the deleted region. The signals of 13q14 and 13qter were separated from each other in 90% of all the primary spermatocytes examined, indicating the unpairing of homologous chromosome 13 or synapse failure. CONCLUSION: Chromosomal rearrangement-induced spermatogenesis failure is caused by the unpairing of the homologous chromosomes involved in the first meiotic division of germ cells.

[Application of Online Mendelian Inheritance in Man to medical genetics].

Online Mendelian Inheritance in Man (OMIM, http://omim. org/) is a comprehensive, authoritative, practical and timely knowledgebase of human genes and genetic disorders. OMIM, as a genetic encyclopedia, provides an easy and straightforward access to information on human genetics to students, researchers and clinicians. This article presents an overview on the contents of OMIM and its application to medical genetics.

[Research progress of Wif1 in development of nervous system].

Wnt inhibitor factor-1 (WIF-1) is an extracellular antagonist of Wnts secreted proteins, first characterized as an expressed sequence tag in the human retina. WIF-1 belongs to the secreted Frizzled-related protein (sFRP) class, which can directly bind to Wnt proteins, prevent Wnts from binding to their receptors in vertebrates. Wif1 is expressed in the nervous system of mouse, Xenopus, zebrafish and human. It has been shown that WIF-1 affects the formation of somites in Xenopus embryos and inhibits rod production in retinal histogenesis by binding to Wnt4 in mice. Histological information of Wif1 expression during the development of the central nervous system has been reported in mouse, Xenopus and zebrafish and the strong embryonic expression suggests Wif1 may play an essential role in the spatial and temporal regulation of Wnt signals in development of central nervous system. The Wnt pathway plays a key role in the patterning of the nervous system. However, insights into the function of Wif1 in the development of the central nervous system are rather limited. Selecting suitable stage and target according to the expression pattern may contribute to understanding the function of Wif1.

A survey on the status of semen analysis in 118 laboratories in China.

Collecting baseline information on how laboratories perform testing is a reasonable first step towards establishing intra- and inter-laboratory standardization and quality control for semen analysis. We carried out a survey of the laboratories performing the testing in Mainland China. A questionnaire, composed of 36 questions covering all aspects of semen analysis, was designed, and a copy was distributed to each of the 145 laboratories. Of these, 118 laboratories completed the questionnaires. The survey results showed that semen volume was measured visually in 53.6% (59/110) of the responding laboratories, and 70.9% (73/103) of laboratories analysed incompletely liquefied semen without any treatment. In addition, both manual-microscopic and computer-assisted semen-analysis systems were applied to analyse sperm concentration, motility and morphology. However, more than five methods were employed in routine sperm staining. An enzyme-linked immunosorbent assay was commonly used for determining whether antisperm antibodies were present. Several seminal biochemical markers were analysed in only 27.1% (32/118) of the responding laboratories. Generally, there was a lack of intra- and inter-laboratory quality control measures for semen analysis in all laboratories responding to this survey. In conclusion, the methods of semen analysis and the interpretation of test results in the surveyed laboratories differed markedly. In particular, many laboratories employed methods other than those recommended by the World Health Organization Laboratory Manual for the Examination of Human Semen and Spermcervical Mucus Interaction (1999). These findings suggest an urgent need for the standardization of semen analysis with acceptable quality controls for each parameter to make the results repeatable and meaningful.

The expression pattern of Follistatin-like 1 in mouse central nervous system development.

Follistatin-like 1 (Fstl1), also named TSC-36 (TGF-beta-stimulated clone 36), was first cloned from the mouse osteoblastic MC3T3-E1 cell line and can be up-regulated by TGF-beta. To better study the function of Fstl1 during the development of the mouse central nervous system (CNS), we examined Fstl1 expression in the developing mouse CNS, in detail, by in situ hybridization. Our results show that Fstl1 is strongly expressed in the telencephalon, diencephalon, brainstem, limbic system and spinal cord. In the telencephalon, Fstl1 positive cells are mainly located in the ventricular zone (VZ) and the subventricular zone (SVZ); a relatively weak signal was observed in layers II and III of the neocortex at postnatal stages. Fstl1 expression is robust in the developing hippocampus and persists to P20. In the developing diencephalon and hindbrain, abundant Fstl1 signals were also detected in nuclei including the medial habenular nucleus, the medial dorsal nucleus, the cochlear nuclei and so on. In addition, a strong expression of Fstl1 was detected in the thalamencephalic signal center, as well as in the olfactory cortex from E14.5 to P0. Meanwhile, Fstl1 was expressed in the septal area and the cingulate gyrus of the limbic system after birth. A high level of expression was also observed in the ventral horn of the spinal cord. These results indicate that Fstl1 may play an important role during CNS development in the mouse.

Expression of Frizzled10 in mouse central nervous system.

Frizzled transmembrane proteins (Fzd) are receptors of Wnts, and they play key roles during central nervous system (CNS) development in vertebrates. Here we report the expression pattern of Frizzled10 in mouse CNS from embryonic stages to adulthood. Frizzled10 is expressed strongly at embryonic days E8.5 and E9.5 in the neural tube and tail bud. At E10.5, Frizzled10 is expressed in the forebrain vesicle, the fourth ventricle and the dorsal spinal cord. From E12.5 to E16.5, Frizzled10 expression is mainly observed in the cortical hem/fimbria, the neuroepithelium of the third ventricular zone, midbrain, developing cerebellum, and dorsal spinal cord. At P0, with the exception of expression in the fimbria, Frizzled10 mRNA expression is limited to specific nuclei including the ventral posterior thalamic nucleus (VP) and the dorsal lateral geniculate nucleus (DLG) in the developing thalamus as well as in the proliferative ventricular zone of the developing cerebellum. From P20 to adult, Frizzled10 mRNA is detected only in the internal capsule (ic). Our data show that expression of Frizzled10 is very strong during embryonic development of the CNS and suggest that Frizzled10 may play an essential role in spatial and temporal regulation during neural development.

Expression pattern of Wnt inhibitor factor 1(Wif1) during the development in mouse CNS.

Wnt inhibitor factor-1 (WIF-1) is an extracellular antagonist of Wnts secreted proteins. Here we describe the expression pattern of Wif1 throughout the development of the mouse central nervous system (CNS). Wif1 mRNA can be detected as early as the developmental stage E11, and expression persists to adulthood. In embryonic stages, the level of Wif1 expression was very prominent in several areas including the cerebral cortex, the diencephalon and the midbrain, with the strongest level in the hippocampal plate and the diencephalon. However, after birth, the expression level of Wif1 decreased in the cortex and diencephalon. By adulthood, Wif1 is mainly expressed in the medial habenular nucleus (MHb) in the epithalamus, the mitral layer cells in the olfactory bulb and a few nuclei in the hypothalamus. Our data shows that the expression of Wif1 was very strong during embryonic development of the CNS and suggests that Wif1 may play an essential role in the spatial and temporal regulation of Wnt signals.

[Evaluation of the determination of seminal ACP and gamma-GT activities and correlation between seminal ACP or gamma-GT activity and semen parameters].

OBJECTIVE: To evaluate the determination of seminal acid phosphatase (ACP) and gamma-glutamyltranspeptidase (gamma-GT) activity, and analyze the correlation between seminal ACP or gamma-GT and semen parameters. METHODS: ACP and gamma-GT activities in 133 samples of seminal plasma were measured. Two of the samples were randomly selected for intra-assay, one for the detection of ACP activity and the other for gamma-GT activity. And another four were selected the same way for the same purpose, two for the detection of ACP activity and the other two for gamma-GT activity. The semen volume, pH, sperm concentration, motility, and grade-a and -b motility were analyzed by CASA system and so were the correlation between seminal ACP or gamma-GT activity and semen parameters. RESULTS: There was significant positive correlation between ACP and gamma-GT activities (r = 0.570, P = 0.000). The intra-CV of ACP was 13.72%, and inter-CVs of ACP were 13.80% and 15.49%. The intra-CV of gamma-GT was 7.68%, and inter-CVs of gamma-GT were 7.76% and 9.73%. Both seminal ACP and gamma-GT activities had significant negative correlation with pH (r = -0.330, P = 0.000 vs r = - 0. 388, P = 0.000). There was obvious correlation between gamma-GT activity and sperm concentration (r = 0.165, P = 0.045), but not between ACP activity and sperm concentration (r = 0.048, P = 0.546). Neither of seminal ACP and gamma-GT activity was correlated with sperm motility, grade-a and -b motility, semen volume, abstinence time and age. CONCLUSION: The precision of the measurement of gamma-GT activity in seminal plasma was higher than that of ACP. The correlation between seminal gamma-GT activity and semen parameters was similar to that between seminal ACP activity and semen parameters. Thus, the determination of gamma-GT activity was a more reliable marker than that of ACP activity for the evaluation of prostate function.