Effect of paternal age on clinical outcomes of in vitro fertilization-embryo transfer cycles.

Purpose: This study aimed to investigate the impact of paternal age > 40 years on clinical pregnancy and perinatal outcomes among patients undergoing in vitro fertilization treatment. Methods: We selected 75 male patients (aged > 40 years) based on predefined inclusion and exclusion criteria. Propensity score matching was performed in a 1:3 ratio, resulting in a control group (aged ≤ 40 years) of 225 individuals. Various statistical tests, including the Mann-Whitney U test, Chi-square test, Fisher's exact test, and binary logistic regression, were used to analyze the association between paternal age and clinical outcomes. Results: We found no statistically significant differences in semen routine parameters, clinical pregnancy outcomes, and perinatal outcomes between paternal aged > 40 and ≤ 40 years. However, in the subgroup analysis, the live birth rate significantly decreased in those aged ≥ 45 compared to those aged 41-42 and 43-44 years (31.25% vs. 69.23% and 65%, respectively; all p < 0.05). Additionally, the clinical pregnancy rate was significantly lower among those aged ≥ 45 than among those aged 41-42 (43.75% vs. 74.36%; p=0.035). Conclusion: Paternal age ≥ 45 years was associated with lower live birth and clinical pregnancy rates.

[Identification of chemical components in Qinggu San reference sample of classial prescription based on UHPLC-Q/Orbitrap HRMS combined with molecular network technology].

The method of ultra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry(UHPLC-Q/Orbitrap HRMS)combined with molecular network was developed in this study for rapidly analyzing the chemical components of the Qinggu San reference sample of classical prescription. Firstly, an ACQUITY UPLC BEH Shield RP_(18) column(2.1 mm×100 mm, 1.7 µm)was used, and acetonitrile and 0.1% formic acid were taken as the mobile phases for gradient elution. The flow rate was 0.4 mL·min~(-1), and the column temperature was 30 ℃. Under these conditions, the mass spectrum data were collected in both positive and negative ion modes of the heated electrospray ionization source. Subsequently, the mass spectrum data of the Qinggu San reference sample were uploaded to the Global Natural Products Social Molecular Network(GNPS)platform for calculation and analysis, and a visual molecular network was built with Cytoscape 3.8.2 software. On this basis, the chemical components of the Qinggu San reference sample were identified by fragmentation regularity of standard compounds, retention time, accurate relative molecular weight of HR-MS, characteristic fragment ions information, literature, and databases. Finally, a total of 105 chemical components were identified and speculated in the Qinggu San reference sample, including 19 iridoid glycosides, 23 flavonoids, 15 phenylpropanoids, 11 triterpene saponins, and 37 other components. Meanwhile, two of these components are potential new compounds. The method used in this study not only achieved rapid and accurate identification of chemical components in the Qinggu San reference sample and provided a scie-ntific basis for the study of pharmacological substances and quality control of Qinggu San compound preparations but also provided a refe-rence for the rapid identification of chemical components in traditional Chinese medicine compound preparations.

[Quality evaluation of Fufang Jinqiancao granules based on ultra performance liquid chromatography-ultraviolet detection quantitative fingerprint combined with chemical pattern recognition].

Fufang Jinqiancao granules have a large market demand due to the fact that they contain diuretics, inhibit urinary calculi formation, and exhibit both anti-inflammatory and antioxidant effects. In the current study, a fast and efficient quantitative ultra performance liquid chromatography-ultraviolet detection (UPLC-UV) fingerprinting method was established to analyze the Fufang Jinqiancao granules, while a chemical pattern recognition technology was used to evaluate the quality of the granules over different years. More specifically, the UPLC-UV system consisted of a Waters Acquity UPLC BEH C18 column (100 mm×2.1 mm, 1.7 µm), acetonitrile (mobile phase A), and a 0.1% formic acid aqueous solution (mobile phase B), wherein a gradient elution protocol was followed. Ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS, Agilent Infinity Ⅱ 1290-6545) was used in combination with reference substances and literature comparisons to identify the common peaks present in the quantitative fingerprint. The fingerprints of 35 batches of Fufang Jinqiancao granules were established by means of the quantitative UPLC-UV fingerprinting method, and the fingerprint data obtained for these samples were further analyzed by chemical pattern recognition techniques, including hierarchical cluster analysis (HCA) and principal component analysis (PCA). The quality difference markers, namely mangiferin and isomangiferin, were screened, and their contents were determined. It was found that 12 common peaks existed in the fingerprint of the Fufang Jinqiancao granules, and the similarities of all 35 batches of samples were greater than 0.952. In addition, for the purpose of HCA, the 35 batches were divided into two categories, of which sample years 2018 and 2019 belonged to one category, and sample years 2020 and 2021 belonged to another category. Notably, PCA gave the same clustering trends as HCA. Based on the obtained results, the mangiferin and isomangiferin components responsible for the differences between the 2018, 2019 and 2020, 2021 samples were further screened by orthogonal partial least squares discriminant analysis (OPLS-DA). Moreover, the contents of the 35 batches of samples were determined using the two differential markers mangiferin and isomangiferin as indicators. The obtained results indicated that the chromatographic peaks of all 35 batches had acceptable resolutions, with mangiferin exhibiting a good linear relationship in the range of 5.3291-133.2276 mg/L, and isomangiferin exhibiting a similar linear relationship in the range of 4.1847-104.6170 mg/L. The average recovery of mangiferin was 101.7%-105.6%, with a relative standard deviation (RSD) of 0.63%-1.43%, while that of isomangiferin was 103.4%-105.5%, with an RSD of 0.60%-1.18%. Importantly, all RSD values were less than 1.43%, thereby indicating that our method meets the requirements of the Chinese Pharmacopoeia (2020 Edition). Among the 35 batches of samples, the contents of mangiferin and isomangiferin were higher in the 2020 and 2021 samples, and the content fluctuation range was smaller. Overall, the development of an accurate and reliable quality control method for Fufang Jinqiancao granules, and a reasonable and effective quality evaluation of Fufang Jinqiancao granule samples from different years was realized. We therefore expect that this study will provide a reference for establishing a more systematic and comprehensive quality control system.

Does ICSI outcome in obstructive azoospermia differ according to the origin of retrieved spermatozoa or the cause of epididymal obstruction? A comparative study.

PURPOSE: To determine whether ICSI outcomes are affected by sperm source or genital tract inflammatory status. METHODS: A retrospective cohort study was conducted in all consecutive obstructive azoospermia patients who underwent testicular sperm aspiration (TESA) or percutaneous epididymal sperm aspiration (PESA) and ICSI between February 1, 2017, and December 31, 2020. Couples were excluded if they were diagnosed with monogenic disease, abnormal karyotype, or had female uterine malformation. The primary objective was to determine whether ICSI outcomes are affected by the use of testicular or epididymal spermatozoa, and the secondary objective was to explore the effect of granulocyte elastase on ICSI outcomes using epididymal spermatozoa. RESULTS: Compared with TESA, inflammatory and non-inflammatory PESA patients exhibited a better high-quality embryo rate, with significant differences among the three groups (49.43 vs. 55.39% and 56.03%; odds ratio, 6.345 and 6.631; 95% confidence interval, 0.340-12.350, and 1.712-11.550; P = 0.038 and P = 0.008, respectively). The fertilization rate, clinical pregnancy rate, live birth delivery rate, and congenital anomaly birth rate were similar in patients who underwent TESA or PESA (with or without inflammation). CONCLUSIONS: The high-quality embryo rate in PESA patients was higher than that in TESA patients. After successful pregnancy, ICSI outcomes did not differ between patients with obstructive azoospermia who experienced TESA or PESA and those with or without genital tract inflammation.

Effects of donor sperm on perinatal and neonatal outcomes resulting from in vitro fertilization-intracytoplasmic sperm injection and embryo transfer cycles: a retrospective cohort study.

Background: The impact of donor sperm on pregnancy outcomes is controversial. The aim of this study was to investigate whether donor sperm in in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) treatment could reduce the rate of live births or increase the incidence of adverse pregnancy outcomes and birth defects in neonates. Methods: This single-centre, retrospective cohort study included 1,559 patients with infertility who received donor sperm at our hospital from 2015 to 2019. All the patients received fresh embryos and underwent first-cycle transfer. After propensity score matching, 4,677 controls who received their partners' sperm were matched at 1:3. Clinical pregnancy, perinatal, and neonatal outcomes were compared between the donor sperm and partner sperm groups. Results: The embryo development was better in the donor sperm group than in the partner sperm group. The high-quality embryo and available embryo rates were significantly higher in the donor sperm group (P<0.05 for both groups). The rate of high-quality embryos transferred from the donor sperm group was higher than that from the partner sperm group (P<0.05). The clinical pregnancy (62.99% vs. 59.65%; P=0.02) and live birth (54.65% vs. 51.59%; P=0.036) rates were higher in the donor sperm group. After adjusting for confounding factors, no significant difference in live birth rates was observed between the two groups (adjusted P=0.057). The low birthweight (18.21% vs. 21.39%; P=0.023) and small for gestational age (SGA) (7.60% vs. 11.97%; P<0.001) rates were lower in the donor sperm group. To exclude the effect of multiple pregnancies, we evaluated neonatal outcomes of singleton pregnancies. No significant differences were noted in preterm and very preterm birth, SGA, mean birthweight, high birthweight, and low birth weight (LBW) and very low birth weight (VLBW) rates (P>0.05 for both groups). Further, no significant between group differences were observed in the ectopic pregnancy rate, early and late spontaneous abortion rates, gestational age, rate of large for gestational age (LGA), and neonatal defects. Conclusions: Compared with partner sperm, donor sperm did not reduce live birth rate and did not increase neonatal LBW or low birth defects.

Rapid Discrimination and Prediction of Ginsengs from Three Origins Based on UHPLC-Q-TOF-MS Combined with SVM.

Ginseng, which contains abundant ginsenosides, grows mainly in the Jilin, Liaoning, and Heilongjiang in China. It has been reported that the quality and traits of ginsengs from different origins were greatly different. To date, the accurate prediction of the origins of ginseng samples is still a challenge. Here, we integrated ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) with a support vector machine (SVM) for rapid discrimination and prediction of ginseng from the three main regions where it is cultivated in China. Firstly, we develop a stable and reliable UHPLC-Q-TOF-MS method to obtain robust information for 31 batches of ginseng samples after reasonable optimization. Subsequently, a rapid pre-processing method was established for the rapid screening and identification of 69 characteristic ginsenosides in 31 batches ginseng samples from three different origins. The SVM model successfully distinguished ginseng origin, and the accuracy of SVM model was improved from 83% to 100% by optimizing the normalization method. Six crucial quality markers for different origins of ginseng were screened using a permutation importance algorithm in the SVM model. In addition, in order to validate the method, eight batches of test samples were used to predict the regions of cultivation of ginseng using the SVM model based on the six selected quality markers. As a result, the proposed strategy was suitable for the discrimination and prediction of the origin of ginseng samples.

Pregnancy outcomes after fresh versus vitrified-warmed embryo transfer in women with adenomyosis: a retrospective cohort study.

RESEARCH QUESTION: Is the singleton live birth rate superior for vitrified-warmed versus fresh embryo transfer in women with adenomyosis? DESIGN: This cohort study retrospectively analysed data from the Reproductive Hospital Affiliated to Shandong University between January 2013 and December 2018. A total of 612 women diagnosed with adenomyosis, with 322 fresh embryo transfer cycles and 290 vitrified-warmed embryo transfer cycles, were included in this study. The primary outcome was singleton live birth. Outcomes were adjusted using multivariable logistic regression analysis. RESULTS: Vitrified-warmed embryo transfer was associated with a higher rate of singleton live birth than fresh embryo transfer (25.9% versus 17.4%; P = 0.011). Although there was a trend towards a lower miscarriage rate after vitrified-warmed embryo transfer, the difference did not reach statistical significance (31.3% versus 40.6%; P = 0.111). The clinical pregnancy rate was comparable in the two groups (44.1% versus 44.4%; P = 0.946). Vitrified-warmed embryo transfer also resulted in a lower risk of preterm birth than fresh embryo transfer (7.0% versus 17.5%; P = 0.010). CONCLUSIONS: Vitrified-warmed embryo transfer may be associated with better pregnancy outcomes than fresh embryo transfer among women with adenomyosis. It seems that vitrified-warmed embryo transfer is more appropriate for specific populations.

Effects of ovarian endometrioma aspiration on in vitro fertilization-intracytoplasmic sperm injection and embryo transfer outcomes: a systematic review and meta-analysis.

PURPOSE: To evaluate the effect of ovarian endometrioma aspiration on IVF/ICSI outcomes. METHODS: The PubMed, EMBASE, Web of Science, China National Knowledge Infrastructure, and Wanfang databases were searched to identify studies related to the treatment of endometrioma up to October 1, 2020, and the data of 1207 patients from 10 studies were analyzed using STATA. RESULTS: The 10 studies in our analysis included 7 comparing aspiration and surgery and 6 comparing aspiration with no intervention. In the aspiration versus surgery groups, live birth rate [OR 0.97 (95% CI 0.51, 1.85), P = 0.925] and clinical pregnancy rate [OR 1.30 (95% CI 0.95, 1.80), P = 0.105] showed no significant difference between the two groups. Abortion rate [OR 4.26 (95% CI 1.38, 13.08), P = 0.011], the number of oocytes retrieved [mean difference 1.95 (95% CI 0.10, 3.81), P = 0.039], and the estradiol peak on hCG day [mean difference 392.16 (95% CI 230.14, 554.18), P < 0.001] were significantly higher in the aspiration group compared to the surgical group. In the aspiration versus the no intervention group, live birth rate [OR 0.84 (95% CI 0.45, 1.59), P = 0.602] and clinical pregnancy rate [OR 1.25 (95% CI 0.88, 1.77), P = 0.206] were not significantly different between the two groups. The abortion rate [OR 0.31 (95% CI 0.11, 0.88), P = 0.028] and the number of gonadotropin ampoules [mean difference - 3.13 ampoules (95% CI - 4.90, - 1.37), P < 0.001] were significantly lower in the aspiration group compared to the no intervention group. CONCLUSION: Compared with surgical treatment or no intervention treatment, aspiration has less effect on ovarian response, ovarian reserve, and pregnancy outcomes.

Increased SPC24 in prostatic diseases and diagnostic value of SPC24 and its interacting partners in prostate cancer.

SPC24 is a crucial component of the mitotic checkpoint machinery in tumorigenesis. High levels of SPC24 have been found in various cancers, including breast cancer, lung cancer, liver cancer, osteosarcoma and thyroid cancer. However, to the best of our knowledge, the impact of SPC24 on prostate cancer (PCa) and other prostate diseases remains unclear. In the present study expression of global SPC24 messenger RNA (mRNA) was assessed in a subset of patients with PCa included in The Cancer Genome Atlas (TCGA) database. Increased levels of SPC24 expression were found in PCa patients >60 years old compared to patients <60 and increased SPC24 expression was also associated with higher levels of prostate specific antigen (P<0.05) and lymph node metastasis (P<0.05). Higher levels of SPC24 expression were associated with negative outcomes in PCa patients (P<0.05). Furthermore, in Chinese patients with prostatitis, benign prostatic hypertrophy (BPH) and PCa, SPC24 was expressed at significantly higher levels than that in adjacent/normal tissues, as assessed by reverse transcription-quantitative polymerase chain reaction, immunohistochemistry and western blotting. High expression of SPC24 was associated with high Gleason stages (IV and V; P<0.05). Further analysis, based on Gene Ontology and pathway functional enrichment analysis, suggested that nuclear division cycle 80 (NDC80), an SPC24 protein interaction partner, and mitotic spindle checkpoint serine/threonine-protein kinase BUB1 (BUB1), a core subunit of the spindle assembly checkpoint, may be associated with SPC24 in PCa development. Finally, using binary logistic regression, algorithms combining the receiver operating characteristic between SPC24 and BUB1 or NDC80 indicated that a combination of these markers may provide better PCa diagnosis ability than other PCa diagnosis markers. Taken together, these findings suggest that SPC24 may be a promising prostate disease biomarker.

Comparison of outcomes in intrauterine insemination, in vitro fertilisation and intracytoplasmic sperm injection in men with and without varicocele.

Objectives: To investigate the differences in clinical pregnancy, miscarriage, and live birth rates when male partners were diagnosed with a varicocele and to compare these outcomes to those without and study the outcomes based on the grade of varicocele. Methods: The retrospective study was based on a cohort of consecutive infertile couples undergoing assisted reproductive technology (ART) at the Reproductive Center of Shandong Provincial Hospital affiliated to the Shandong University during the period between January 2017 and December 2018. A total of 4203 couples comprised of men with and without varicocele undergoing the first ART cycle (1501 intrauterine inseminations (IUI), 1623 in vitro fertilisations (IVF) and 1079 intracytoplasmic sperm injections (ICSI)) were included. Semen parameters and ART outcomes were determined. Results: ICSI (26.5%) originated from men with a significant lower level in sperm concentration and motility but with a strict normal morphology had a higher prevalence of varicocele than men undergoing IUI (20.7%) and IVF (18.1%). In IUI, the odds ratios (ORs) for pregnancy and live birth were significantly lower for couples in men diagnosed with grades 1 or 2 varicocele as compared to those for men with grade 3 varicocele. In IVF, ORs for live birth where men were diagnosed with grades 1 or 2 varicocele were also lower than those for men with grade 3,whereas a higher miscarriage rate was found when men had grades 1 or 2 varicocele than when men had grade 3. However, for ICSI, no significant outcomes were found in grades 1, 2 or 3 varicocele versus the no varicocele group. Conclusions: The increasing grade of varicocele was negatively associated with sperm parameters and can alter the outcome of further IUI/IVF.

Success rates of in vitro fertilization versus intracytoplasmic sperm injection in men with serum anti-sperm antibodies: a consecutive cohort study.

Antisperm antibodies (ASAs) are assumed to be a possible causative factor for male infertility, with ASAs detected in 5%-15% of infertile men but in only 1%-2% of fertile ones. It remains unclear whether ASAs have an adverse effect on the outcome of in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). This study investigated differences in the rates of fertilization, pregnancy, and live births associated with serum ASA-positive and ASA-negative men following IVF or ICSI. Five hundred and fifty-four consecutive infertile couples undergoing IVF (n = 399) or ICSI (n = 155) were included. The two-sample two-sided t-test and Chi-square or Fisher's exact test was used for statistical analysis. Lower rates of fertilization (41.7% vs 54.8%, P = 0.03), good embryos (18.9% vs 35.2%, P = 0.00), pregnancy (38.5% vs 59.4%, P = 0.00), and live births (25.8% vs 42.5%, P = 0.00) were observed in men of the IVF group with a positive serum ASA than in those with a negative ASA. ASA positivity/negativity correlated with pregnancy rates (P = 0.021, odds ratio [OR]: 0.630, 95% confidence interval [CI]: 0.425-0.932) and live birth rates (P = 0.010, OR: 1.409, 95% CI: 1.084-1.831) after controlling for the female serum follicle-stimulating hormone level and the couple's ages at IVF. Women coupled with ASA-positive men had lower live birth rates with IVF than with ICSI (25.8% and 47.4%, respectively; P = 0.07). Women coupled with ASA-positive men had lower rates of pregnancy and live births following IVF than those coupled with ASA-negative men but had a similar outcome with ICSI.

Human leukocyte antigen (HLA)-DRB1 allele polymorphisms and systemic sclerosis.

Objectives: Human leukocyteantigen (HLA) is the most important gene for immune system regulation. Although studies have evaluated the association between HLA-DRB1 allele polymorphisms and systemic sclerosis (SSc), their results are still controversial. We performed a meta-analysis to assess the association of HLA-DRB1 alleles with risk of SSc.Methods: Electronic database were systematically searched for articles, a total of 11 case-control studies including 3268 cases and 5548 controls were analyzed. Odds ratio (ORs) and 95% confidence intervals were used to assess the association of HLA-DRB1 alleles with SSc. The relationship between SSc-related autoantibodies and DRB1 alleles was also analyzed.Results: In the overall analysis, four alleles (DRB1*04:03, DRB1*08, DRB1*11, and DRB1*11:04) increased the risk of SSc; however, five alleles (DRB1*07, DRB1*11:01, DRB1*13, DRB1*13:01, and DRB1*14) had the opposite effect. Analysis of subgroups by ethnicity indicate that DRB1*11:01 and DRB1*13:01 confer a protective effect in Caucasians, while DRB1*11:04 was associated with a higher risk of SSc. For Asian, DRB1*13:02 was found to be a protective factor. In addition, the frequency of DRB1*11:04 alleles was significantly increased in ATA+ SSc patients compared with ATA- SSc patients.Conclusion: DRB1*04:03, DRB1*08, DRB1*11, and DRB1*11:04 were associated with the risk of SSc. Additionally, DRB1*11 and DRB1*11:04 were association with ATAs.

HIV Tat causes synapse loss in a mouse model of HIV-associated neurocognitive disorder that is independent of the classical complement cascade component C1q.

Microglial activation, increased proinflammatory cytokine production, and a reduction in synaptic density are key pathological features associated with HIV-associated neurocognitive disorders (HAND). Even with combination antiretroviral therapy (cART), more than 50% of HIV-positive individuals experience some type of cognitive impairment. Although viral replication is inhibited by cART, HIV proteins such as Tat are still produced within the nervous system that are neurotoxic, involved in synapse elimination, and provoke enduring neuroinflammation. As complement deposition on synapses followed by microglial engulfment has been shown during normal development and disease to be a mechanism for pruning synapses, we have tested whether complement is required for the loss of synapses that occurs after a cortical Tat injection mouse model of HAND. In Tat-injected animals evaluated 7 or 28 days after injection, levels of early complement pathway components, C1q and C3, are significantly elevated and associated with microgliosis and a loss of synapses. However, C1qa knockout mice have the same level of Tat-induced synapse loss as wild-type (WT) mice, showing that the C1q-initiated classical complement cascade is not driving synapse removal during HIV1 Tat-induced neuroinflammation.

Corrigendum: Platelet Activating Factor Enhances Synaptic Vesicle Exocytosis Via PKC, Elevated Intracellular Calcium, and Modulation of Synapsin 1 Dynamics and Phosphorylation.

[This corrects the article on p. 505 in vol. 9, PMID: 26778968.].

Liquid nitrogen vapor is comparable to liquid nitrogen for storage of cryopreserved human sperm: evidence from the characteristics of post-thaw human sperm.

OBJECTIVE: To compare the differences in the characteristics of post-thaw human sperm after storage in either liquid nitrogen (LN2; -196 °C) or LN2 vapor (-167 °C). DESIGN: Experimental study. SETTING: University hospital. PATIENT(S): Thirty healthy volunteers who agreed to donate their normal semen samples for infertility or research were included in the study. INTERVENTION(S): Semen samples (n = 30) were divided into eight aliquots and frozen. Four aliquots of each human semen sample were stored in LN2 (-196 °C), and the other four aliquots were stored in LN2 vapor (-167 °C). After 1, 3, 6, or 12 months, samples were thawed and analyzed. MAIN OUTCOME MEASURE(S): The motility was evaluated by the manual counting method. The viability was estimated by eosin staining. The morphology was analyzed by Diff-Quik staining. The sperm DNA integrity was determined with acridine orange fluorescent staining, and acrosin activity was assayed by the modified Kennedy method. RESULT(S): The characteristics of post-thaw human sperm, including motility, viability, morphology, DNA integrity, and acrosin activity, showed no significant difference between LN2 and LN2 vapor storage for the different time periods. CONCLUSION(S): LN2 vapor was comparable to LN2 in post-thaw sperm characteristics, suggesting that LN2 vapor may be substituted for LN2 for the long-term storage of human sperm.

Serum quantitative proteomic analysis reveals potential zinc-associated biomarkers for nonbacterial prostatitis.

BACKGROUND: Prostatitis is one of the most common urological problems afflicting adult men. The etiology and pathogenesis of nonbacterial prostatitis, which accounts for 90-95% of cases, is largely unknown. As serum proteins often indicate the overall pathologic status of patients, we hypothesized that protein biomarkers of prostatitis might be identified by comparing the serum proteomes of patients with and without nonbacterial prostatitis. METHODS: All untreated samples were collected from subjects attending the Fangchenggang Area Male Health and Examination Survey (FAMHES). We profiled pooled serum samples from four carefully selected groups of patients (n = 10/group) representing the various categories of nonbacterial prostatitis (IIIa, IIIb, and IV) and matched healthy controls using a mass spectrometry-based 4-plex iTRAQ proteomic approach. More than 160 samples were validated by ELISA. RESULTS: Overall, 69 proteins were identified. Among them, 42, 52, and 37 proteins were identified with differential expression in Category IIIa, IIIb, and IV prostatitis, respectively. The 19 common proteins were related to immunity and defense, ion binding, transport, and proteolysis. Two zinc-binding proteins, superoxide dismutase 3 (SOD3), and carbonic anhydrase I (CA1), were significantly higher in all types of prostatitis than in the control. A receiver operating characteristic curve estimated sensitivities of 50.4 and 68.1% and specificities of 92.1 and 83.8% for CA1 and SOD3, respectively, in detecting nonbacterial prostatitis. The serum CA1 concentration was inversely correlated to the zinc concentration in expressed-prostatic secretions. CONCLUSIONS: Our findings suggest that SOD3 and CA1 are potential diagnostic markers of nonbacterial prostatitis, although further large-scale studies are required. The molecular profiles of nonbacterial prostatitis pathogenesis may lay a foundation for discovery of new therapies.

Leucine-rich repeat kinase 2 modulates neuroinflammation and neurotoxicity in models of human immunodeficiency virus 1-associated neurocognitive disorders.

Leucine-rich repeat kinase 2 (LRRK2) is the single most common genetic cause of both familial and sporadic Parkinson's disease (PD), both of which share pathogenetic and neurologic similarities with human immunodeficiency virus 1 (HIV-1)-associated neurocognitive disorders (HAND). Pathologic LRRK2 activity may also contribute to neuroinflammation, because microglia lacking LRRK2 exposed to proinflammatory stimuli have attenuated responses. Because microglial activation is a hallmark of HIV-1 neuropathology, we have investigated the role of LRRK2 activation using in vitro and in vivo models of HAND. We hypothesize that LRRK2 is a key modulator of microglial inflammatory responses, which play a pathogenic role in both HAND and PD, and that these responses may cause or exacerbate neuronal damage in these diseases. The HIV-1 Tat protein is a potent neurotoxin produced during HAND that induces activation of primary microglia in culture and long-lasting neuroinflammation and neurotoxicity when injected into the CNS of mice. We found that LRRK2 inhibition attenuates Tat-induced pS935-LRRK2 expression, proinflammatory cytokine and chemokine expression, and phosphorylated p38 and Jun N-terminal kinase signaling in primary microglia. In our murine model, cortical Tat injection in LRRK2 knock-out (KO) mice results in significantly diminished neuronal damage, as assessed by microtubule-associated protein 2 (MAP2), class III ß-tubulin TUJ1, synapsin-1, VGluT, and cleaved caspase-3 immunostaining. Furthermore, Tat-injected LRRK2 KO animals have decreased infiltration of peripheral neutrophils, and the morphology of microglia from these mice were similar to that of vehicle-injected controls. We conclude that pathologic activation of LRRK2 regulates a significant component of the neuroinflammation associated with HAND.

Platelet Activating Factor Enhances Synaptic Vesicle Exocytosis Via PKC, Elevated Intracellular Calcium, and Modulation of Synapsin 1 Dynamics and Phosphorylation.

Platelet activating factor (PAF) is an inflammatory phospholipid signaling molecule implicated in synaptic plasticity, learning and memory and neurotoxicity during neuroinflammation. However, little is known about the intracellular mechanisms mediating PAF's physiological or pathological effects on synaptic facilitation. We show here that PAF receptors are localized at the synapse. Using fluorescent reporters of presynaptic activity we show that a non-hydrolysable analog of PAF (cPAF) enhances synaptic vesicle release from individual presynaptic boutons by increasing the size or release of the readily releasable pool and the exocytosis rate of the total recycling pool. cPAF also activates previously silent boutons resulting in vesicle release from a larger number of terminals. The underlying mechanism involves elevated calcium within presynaptic boutons and protein kinase C activation. Furthermore, cPAF increases synapsin I phosphorylation at sites 1 and 3, and increases dispersion of synapsin I from the presynaptic compartment during stimulation, freeing synaptic vesicles for subsequent release. These findings provide a conceptual framework for how PAF, regardless of its cellular origin, can modulate synapses during normal and pathologic synaptic activity.

Effect of trichostatin A on CNE2 nasopharyngeal carcinoma cells--genome-wide DNA methylation alteration.

Trichostatin A (TSA) is a histone deacetylase (HDAC) inhibitor. We here investigated its effects on proliferation and apoptosis of the CNE2 carcinoma cell line, and attempted to establish genome-wide DNA methylation alteration due to differentially histone acetylation status. After cells were treated by TSA, the inhibitory rate of cell proliferation was examined with a CCK8 kit, and cell apoptosis was determined by flow cytometry. Compared to control, TSA inhibited CNE2 cell growth and induced apoptosis. Furthermore, TSA was found to induce genome-wide methylation alteration as assessed by genome-wide methylation array. Overall DNA methylation level of cells treated with TSA was higher than in controls. Function and pathway analysis revealed that many genes with methylation alteration were involved in key biological roles, such as apoptosis and cell proliferation. Three genes (DAP3, HSPB1 and CLDN) were independently confirmed by quantitative real-time PCR. Finally, we conclude that TSA inhibits CNE2 cell growth and induces apoptosis in vitro involving genome-wide DNA methylation alteration, so that it has promising application prospects in treatment of NPC in vivo. Although many unreported hypermethylated/hypomethylated genes should be further analyzed and validated, the pointers to new biomarkers and therapeutic strategies in the treatment of NPC should be stressed.

Association of cystic fibrosis transmembrane-conductance regulator gene mutation with negative outcome of intracytoplasmic sperm injection pregnancy in cases of congenital bilateral absence of vas deferens.

OBJECTIVE: To evaluate intracytoplasmic sperm injection (ICSI) results with regard to congenital bilateral absence of vas deferens (CBAVD) versus non-CBAVD obstruction, cystic fibrosis transmembrane-conductance regulator (CFTR) mutations versus non-CFTR mutations, and miscarriages or stillbirths versus live births per embryo transferred. DESIGN: Retrospective study with detailed chart review. SETTING: Center for reproductive medicine. PATIENT(S): Nine hundred forty-five men with obstructive azoospermia. INTERVENTION(S): One thousand four hundred fourteen ICSI cycles classified as CBAVD versus non-CBAVD obstruction, CFTR mutations versus non-CFTR mutations, and miscarriages/stillbirths versus live births per embryo transferred. MAIN OUTCOME MEASURE(S): Frequency of CFTR mutations and rates of fertilization, good embryos, clinical pregnancy, miscarriages and stillbirths, ectopic pregnancy, and live births. RESULT(S): CFTR mutations were more prevalent in men with CBAVD than in those with non-CBAVD obstruction. The rate of miscarriages and stillbirths per embryo transferred was higher in men with CBAVD than in those with non-CBAVD obstruction, whereas the rate of live births per embryo transferred was lower in men with CBAVD than in those with non-CBAVD obstruction. The rate of miscarriages and stillbirths per embryo transferred was higher in men with CFTR mutations than in those with non-CFTR mutations. The frequency of CFTR mutations was higher in patients who experienced miscarriages/stillbirths than in those with live births. CONCLUSION(S): The frequency of CFTR mutations was higher in cases of CBAVD versus non-CBAVD obstruction. Possibly as a result of CFTR mutations, patients with CBAVD had a significantly increased risk of miscarriage and stillbirth and a reduced rate of live birth compared with patients with non-CBAVD.