Abstract Background Spermatogonial stem cells (SSCs) are critical for sustaining spermatogenesis. Even though several regulators of SSC have been identified in rodents, the regulatory mechanism of SSC in humans has yet to be discovered. Methods To explore the regulatory mechanisms of human SSCs, we analyzed publicly available human testicular single-cell sequencing data and found that Ankyrin repeat and SOCS box protein 9 (ASB9) is highly expressed in SSCs. We examined the expression localization of ASB9 using immunohistochemistry and overexpressed ASB9 in human SSC lines to explore its role in SSC proliferation and apoptosis. Meanwhile, we used immunoprecipitation to find the target protein of ASB9 and verified its functions. In addition, we examined the changes in the distribution of ASB9 in non-obstructive azoospermia (NOA) patients using Western blot and immunofluorescence. Results The results of uniform manifold approximation and projection (UMAP) clustering and pseudotime analysis showed that ASB9 was highly expressed in SSCs, and its expression gradually increased during development. The immunohistochemical and dual-color immunofluorescence results displayed that ASB9 was mainly expressed in nonproliferating SSCs. Overexpression of ASB9 in the SSC line revealed significant inhibition of cell proliferation and increased apoptosis. We predicted the target proteins of ASB9 and verified that hypoxia-inducible factor 1-alpha inhibitor (HIF1AN), but not creatine kinase B-type (CKB), has a direct interaction with ASB9 in human SSC line using protein immunoprecipitation experiments. Subsequently, we re-expressed HIF1AN in ASB9 overexpressing cells and found that HIF1AN reversed the proliferative and apoptotic changes induced by ASB9 overexpression. In addition, we found that ABS9 was significantly downregulated in some NOA patients, implying a correlation between ASB9 dysregulation and impaired spermatogenesis. Conclusion ASB9 is predominantly expressed in human SSCs, it affects the proliferation and apoptotic process of the SSC line through HIF1AN, and its abnormal expression may be associated with NOA.

Abstract Objective Abnormal complement activation is associated with periodontitis. W54011 is a novel non-peptide C5aR antagonist (C5aRA) that exhibits favorable anti-inflammatory effects in various inflammatory models. However, whether W54011 inhibits periodontitis has not yet been fully elucidated. To address this, we have investigated the probable anti-inflammatory mechanism of W54011 in LPS-treated inflammation in human gingival fibroblasts (HGFs). Methodology HGFs were isolated from healthy gingival tissue samples using the tissue block method and were identified with immunofluorescence staining. The CCK8 assay and reverse transcription-PCR (RT-PCR) were used to select the optimal induction conditions for Lipopolysaccharide (LPS) and C5aRA (according to supplementary data S1, S2 and S3). The levels of inflammatory cytokines, C5aR, and the activation of NF-κB/MAPK signaling pathways were determined by RT-quantitative PCR (RT-qPCR) and Western blotting. Results Immunofluorescence results showed that vimentin and FSP-1 were positive in HGFs and Keratin was negative in HGFs. Immunofluorescence staining demonstrated that C5aRA inhibited LPS-stimulated nuclear translocation of p-p65. RT-qPCR and Western blotting showed that C5aRA reduced the expression of IL-1β, IL-6, TNF-α, C5aR, p-p65, p-IκBα, p-JNK, p-c-JUN, and TLR4 in LPS-induced HGFs. Conclusion These findings suggested that C5aRA attenuated the release of inflammatory cytokines in LPS-induced HGFs by blocking the activation of the NF-κB and MAPK signaling pathways.

Abstract Atrial fibrillation (AF) represents the most common type of sustained cardiac arrhythmia in humans and confers a significantly increased risk for thromboembolic stroke, congestive heart failure and premature death. Aggregating evidence emphasizes the predominant genetic defects underpinning AF and an increasing number of deleterious variations in more than 50 genes have been involved in the pathogenesis of AF. Nevertheless, the genetic basis underlying AF remains incompletely understood. In the current research, by whole-exome sequencing and Sanger sequencing analysis in a family with autosomal-dominant AF and congenital patent ductus arteriosus (PDA), a novel heterozygous variation in the PRRX1 gene encoding a homeobox transcription factor critical for cardiovascular development, NM_022716.4:c.373G>T;p.(Glu125*), was identified to be in co-segregation with AF and PDA in the whole family. The truncating variation was not detected in 306 unrelated healthy individuals employed as controls. Quantitative biological measurements with a reporter gene analysis system revealed that the Glu125*-mutant PRRX1 protein failed to transactivate its downstream target genes SHOX2 and ISL1, two genes that have been causally linked to AF. Conclusively, the present study firstly links PRRX1 loss-of-function variation to AF and PDA, suggesting that AF and PDA share a common abnormal developmental basis in a proportion of cases.

Abstract As the main signal for the maternal recognition in ruminants, interferon-tau (IFNT) stimulates expression of interferon-stimulated genes (ISGs) in uterus and many extrauterine tissues. However, it is unclear that early pregnancy induces expression of signal transducer and activator of transcription 1 (STAT1), myxovirusresistance 1 (Mx1), interferon-gamma-inducible protein 10 (IP-10) and ubiquitin activating enzyme E1-like protein (UBE1L) in maternal thymus. In this study, ovine thymuses were sampled on day 16 of the estrous cycle and on days 13, 16 and 25 of gestation, and the expression of STAT1, Mx1, IP-10 and UBE1L was detected by real-time quantitative PCR, Western blot and immunohistochemistry. The results revealed that the expression of STAT1 and IP-10 reached peaks on day 16 of pregnancy, and expression of Mx1 was enhanced on day 25 of pregnancy, and STAT1 protein was located in the epithelial reticular cells, capillaries and thymic corpuscles. However, expression of UBE1L was declined during early pregnancy. In conclusion, early pregnancy influences expression of STAT1, Mx1, IP-10 and UBE1L in maternal thymus, which may participate in regulation of maternal immune tolerance during early pregnancy in sheep.

Abstract Liver plays important roles in the innate and adaptive immunity, and contributes to the maternal immune adjustments during pregnancy in mice and rats. T helper 1 (Th1) and Th2 cytokines are related to immune response. However, expression of Th1 and Th2 cytokines in maternal livers is unclear during early pregnancy in sheep. In this study, livers were collected on day 16 of the estrous cycle and on days 13, 16 and 25 of pregnancy (n = 6 for each group) in ewes, and qRT-PCR, western blot and immunohistochemistry were used to analyze the expression of Th1 and Th2 cytokines in the livers. Our results showed that interferon-gamma (IFN-γ), interleukin (IL)-2, IL-4, IL-6 and IL-10 were downregulated, and IL-5 was upregulated in the livers during early pregnancy. Furthermore, there was no effect for early pregnancy on expression of TNF-β in the livers, and the IFN-γ protein was limited to the endothelial cells of the proper hepatic arteries and portal veins. In conclusion, early pregnancy exerted its effect on the liver to regulate the Th cytokines expression, but there was no evident shift from Th1 to Th2 cytokines, which may be necessary for the maternal hepatic immune adjustments during early pregnancy in sheep.

ABSTRACT A new saurolophine hadrosaurid, Laiyangosaurus youngi gen. et sp. nov. is described and phylogenetically analyzed based on several cranial elements from the Jingangkou Formation, Wangshi Group, Upper Cretaceous of Laiyang, Shandong, China. Laiyangosaurus youngi differs from other members of the saurolophine clade on the basis of a number of autapomorphies, including a prominent and narrow ridge on the lateral side of the nasal which forms the posterodorsal and posterior margin of the circumnasal depression, a primary ridge that runs along most of the maxillary tooth row that is slightly deflected posteriorly, a retroarticular process of the surangular that is dorsolateroposteriorlly recurved, and orbital margins that are wider than the infratemporal margins of the jugal. This new taxon can be further distinguished by a number of unique combination of characters, including dorsal margin of nasal is flat, absence supracranial crest, a relatively shallow and rostrodorsally directed caudal margin of the lacrimal process of the jugal, and one or more foramina present on the rostral surface of the premaxilla. A phylogenetic analysis indicates that L. youngi comprises a monophyletic clade, which is known as Edmontosaurini.

Abstract: Chuju, the flower of Chrysanthemum morifolim cv. Chuju, is a traditional medicine-food material used in China. Bioassay-guided chemical investigation on Chuju led the isolation of 16 flavonoids (1-16) characterized as quercetin (1), isorhamnetin 3-O-b-D-glucoside (2), eriodictyol (3), pyracanthoside (4), apigenin (5), apigetrin (6), acacetin (7), acacipetalin (8), luteolin (9), diosmetin (10), spinacetin (11), axillarin (12), bonanzin (13), cirsiliol (14), chrysosplenol D (15) and artemetin (16). Among them, compounds (11-16) were first reported from the Chrsanthemum species. Chuju flavonoids displayed strong antioxidant in DPPH (1, 1-diphenyl-2- picrylhydrazyl) radical scavenging assay, and high anti-inflammatory activity by inhibiting NO production in LPS-induced RAW 264.7 cells. These findings suggests that Chuju is an ideal source of natural flavonoids with significant antioxidant and anti-inflammatory effects, and is worthy of further development as health-promoting food or supplement.

Resumen: La flor de Chrysanthemum morifolim cv. Chuju, conocida popularmente como Chuju, se utiliza tradicionalmente como alimento y medicina en China. Un estudio químico biodirigido de un extracto de las flores condujo al aislamiento de 16 flavonoides, los cuales se caracterizaron como quercetina (1), isoramnetina 3-O-b-D-glucosido (2), eriodictiol (3), piracantosido (4), apigenina (5), apigetrina (6), acacetina (7), acacipetalina (8), luteolina (9), diosmetina (10), espinacetina (11), axilarina (12), bonanzina (13), cirsiliol (14), crisosplenol D (15) and artemetina (16). Los productos 11-16 se describen por primera vez en la especie. Los flavonoides de Chuju demostraron una notable actividad antioxidante en el ensayo de captación del radical 2,2- difenil-1-picrilhidrazilo (DPPH); asimismo presentaron un fuerte efecto antiinflamatorio in vitro, valorado a través de la inhibición de la producción de NO en macrófagos RAW 264.7 estimulados con LPS. Estos resultados indican que Chuju es una fuente importante de flavonoides antiinflamatorios y antioxidantes con un potencial importante para su desarrollo como agente nutracéutico.

This part inquires the presence of mental health disorders in Chinese immigrants (coolies) who ca me to Peru between 1849 and 1874 as indentured laborers to supply the lack of manpower after the abolition of slavery. Historical publications about how this population lived were reviewed. The main reports were based on historical records from labor places between 1865 and 1900 and let us outline emotional and conduct disorders related to sustained stress (traumatic draft, long and unhealthy sea voyage, life without family, enslaved work and confinement in slave quarters). It is inferred the presence of asthenic states due to somatic diseases, depression, hypochondriasis or simulation in workers called 'mañosos' (tricky). Literature refers homosexual practices in this male population in compulsory confinement and sexually transmitted diseases including syphilis; a great consume of opium, not replaced by alcohol or coca; hopelessness; resistance and revolt attitudes; suicides; homicides; group revolts and collaborationism during the Pacific War. These historical findings confronted with hospital statistics of the Manicomio del Cercado (part 1) coincidently point out high opium consume, diagnosed in the hospital as 'toxic psychosis' associated with this narcotic. There is a discrepancy between historical reports of frequent sexually transmitted diseases, including syphilis and the low hospital prevalence of general paresis, the psychiatric form of neurosyphilis. This incongruence is discussed and the possibility of an ecological malariotherapy as an influencing factor on the low hospital prevalence of general paresis is proposed.

Se indagó sobre los desórdenes de la salud mental de los inmigrantes chinos (culíes) que llegaron al Perú en el siglo XIX, entre 1849 y 1874, para suplir la carencia de mano de obra después de la abolición de la esclavitud. Se revisó las publicaciones históricas sobre las condiciones de vida de esa población; los datos se sustentan en registros provenientes de sus centros de trabajo. Los informes, que abarcan el período entre 1865 y 1900, permitieron delinear alteraciones emocionales y de conducta relacionadas con estrés prolongado (reclutamiento traumático, larga e insalubre travesía marítima, sin familia, con trabajo en condiciones de semiesclavitud y encierro en galpones). Se colige la presencia de estados asteniformes somatógenos, depresivos, hipocondríacos o de simulación en los trabajadores calificados como 'mañosos'. La literatura refiere prácticas homosexuales en el encierro obligado de varones solteros y enfermedades de transmisión sexual que incluye a la sífilis; un elevado consumo de opio que trató de ser reemplazado por alcohol y coca; actitudes de desesperanza, resistencia y rebeldía que condujeron a suicidios y homicidios así como a rebeliones grupales y a colaboracionismo durante la Guerra del Pacífico. Estos hallazgos históricos, confrontados con las estadísticas hospitalarias del Manicomio del Cercado (primera parte), coinciden en señalar el alto consumo de opio, por los cuadros diagnosticados en el hospital psiquiátrico como 'locura tóxica' asociada a ese estupefaciente. Hay una discrepancia entre los informes históricos de frecuentes enfermedades de transmisión sexual, incluyendo sífilis, y la escasa prevalencia hospitalaria de parálisis general progresiva (PGP), la forma psiquiátrica de la neurosífilis revelada en la primera parte de este estudio. En la discusión de esta incongruencia se plantea la posibilidad de una malarioterapia ecológica como factor influyente en la baja prevalencia hospitalaria de PGP.

A retrospective study was made in order to determine a psychiatric statistical profile of the hospitalized Chinese immigrants ('coolies') who came to Peru in the XIX century as indentured labourers alter the abolition of slavery. In the archives of the Manicomio del Cercado in Lima, guarded in the Hospital Victor Larco Herrera; 58 hospitalized coolies were registered between 1859 and 1917, all of them male, mainly single and elder than 40 years old. The most frequent diagnosis (57%) was 'toxic psychosis', 47% associated to opium and 10% to alcohol and coca leaves consume. The rest of the diagnosis could be equivalent to schizophrenia or dementia and bipolar disorders and fewer to paranoia and epileptic psychosis. Only one patient (1.7%) was diagnosed as 'toxic psychosis' associated to syphilis: general paresis of the insane, the psychiatric form of neurosyphilis, whose frequency was even ten times more in other countries.

Para determinar un perfil estadístico hospitalario de los desórdenes psiquiátricos de los inmigrantes chinos ('culíes') que llegaron al Perú en el siglo XIX (entre 1849 y 1874), después de la abolición de la esclavitud, se realizó un estudio retrospectivo en los archivos del Manicomio del Cercado de Lima conservados en el Hospital Víctor Larco Herrera. Se halló que, entre 1879 y 1902, estuvieron hospitalizados 58 culíes, todos varones, predominantemente solteros y mayores de 40 años. El 57% de los diagnósticos corresponde a 'psicosis tóxica' asociada, fundamentalmente, a opio, 47%, y el10% restante a consumo de alcohol y coca. Los otros diagnósticos podrían ser equivalentes a esquizofrenia o demencia y trastornos bipolares; más escasos, paranoia y psicosis epiléptica. Solo un paciente (1,7%) tuvo el diagnóstico de 'psicosis tóxica' asociada a sífilis ('parálisis general progresiva' (PGP), la forma psiquiátrica de la neurosífilis, cuya frecuencia era hasta diez veces más en otros países.

Abstract: A new sesquilignan effususin E (1) and a new long chain fatty enamide effususin F (2), together with eight known compounds were isolated from the medullae of Juncus effusus. The structures of 1 and 2 were elucidated by detailed spectroscopic analysis including 1H and 13C NMR, COSY, HSQC, HMBC, and HRMS, while the structures of the known compounds were deduced from comparison of their spectral data with those in the literature. The new compounds 1 and 2 were evaluated against five human cancer cell lines (SHSY-5Y, SMMC-7721, HepG-2, Hela, and MCF-7) by CCK-8 assay, but only 1 exhibited weak cytotoxic activity against SMMC-7721 with IC50 value of 57.5 μm. Inhibitory effects of 1 and 2 on nitric oxide (NO) production in lipopolysaccharide-induced RAW264.7 macrophages were also evaluated, and they were found to exhibit good inhibitory activities with IC50 values of 8.59 and 13.73 μm, respectively.

Resumen: Se elucidaron las estructuras de 1 y 2 mediante análisis espectrocópicos de RMN de 1H y 13C, COSY, HSQC, HMBC, y HRMS, mientras que las estructuras de los compuestos conocidos se dedujeron por comparación de sus datos espectrales con los de la literatura. Los nuevos compuestos 1 y 2 fueron evaluados contra cinco líneas celulares cancerígenas humanas (SHSY-5Y, SMMC-7721, HepG-2, Hela, and MCF-7) mediante el ensayo CCK-8. Solamente el compuesto 1 mostró una débil actividad citotóxica contra la línea SMMC-7721 con un valor de IC50 de 57.5 μM. También se evaluó el efecto inhibitorio de los compuestos 1 y 2 en la producción de óxido nítrico (NO) en macrófagos RAW264.7 inducidos por lipopolisacáridos. Ambos compuestos mostraron buena actividad inhibitoria con valores de IC50 de 8.59 y 13.73 μM, respectivamente.

OBJECTIVE: Patients with nasopharyngeal carcinoma experience highly variable outcomes despite receiving similar therapeutic regimens. Identifying biomarkers that predict survival and guide individualized therapy is urgently needed. Cystatin C has been explored as a valuable prognostic marker in several malignancies. We retrospectively assessed the relationship between serum cystatin C levels and nasopharyngeal carcinoma prognosis in a large cohort of nasopharyngeal carcinoma patients receiving long-term follow-up. METHODS: A total of 1063 consecutive patients diagnosed with nasopharyngeal carcinoma from June 2006 to December 2010 were retrospectively analyzed. The serum levels of cystatin C at the time of diagnosis were collected. Receiver operating characteristic curve analysis, the Kaplan-Meier method and multivariate analyses using a Cox regression model were performed to assess the correlation of cystatin C levels with overall survival, progression-free survival, distant metastasis-free survival and loco-regional recurrence-free survival. RESULTS: The median follow-up duration was 68.3 months. The optimal cut-off value of cystatin C levels for predicting death was 0.945 mg/L. Compared with the low cystatin C group, the high cystatin C group experienced significantly shorter overall survival (hazard ratio=1.47, p=0.050), progression-free survival (hazard ratio=1.65, p=0.004), distant metastasis-free survival (hazard ratio=2.37, p<0.001) and loco-regional recurrence-free survival (hazard ratio=2.40, p=0.002). Based on multivariate analysis, a high cystatin C level was identified as a significant and independent negative predictor of overall survival (hazard ratio=1.47, p=0.050), progression-free survival (hazard ratio=1.65, p=0.004), distant metastasis-free survival (hazard ratio=2.37, p<0.001), and loco-regional recurrence-free survival (hazard ratio=2.40, p=0.002). CONCLUSION: Cystatin C levels are associated with the prognosis of nasopharyngeal carcinoma patients. A high cystatin C level is an independent indicator of poor prognosis for nasopharyngeal carcinoma patients.

A contraparte de um espécime anteriormente descrito, contendo uma fêmea de pterossauro não-pterodactiloide com um ovo, revela a existência de um segundo ovo, este situado dentro da cavidade corporal desta fêmea prenhe. O exemplar demonstra claramente que os pterossauros possuíam dois ovidutos funcionais, e evidencia que a redução de um oviduto não foi um pré-requisito para o desenvolvimento do voo ativo pelo menos neste grupo. A análise da composição de um dos ovos sugere a ausência de uma camada externa formada por carbonato de cálcio. Seções histológicas de um fêmur não registram osso medular e demonstram que este pterossauro atingiu maturidade reprodutiva antes da maturidade óssea. O presente estudo evidencia que os pterossauros punham ovos de tamanho ainda menor do que se supunha e que desenvolveram uma estratégia reprodutiva mais similar a dos répteis basais do que a das aves. Se pterossauros eram altamente precociais ou necessitavam de cuidado parental ainda está aberto à discussão.

The counterpart of a previously described non-pterodactyloid pterosaur with an egg revealed the presence of a second egg inside the body cavity of this gravid female. It clearly shows that pterosaurs had two functional oviducts and demonstrates that the reduction of one oviduct was not a prerequisite for developing powered flight, at least in this group. Compositional analysis of one egg suggests the lack of a hard external layer of calcium carbonate. Histological sections of one femur lack medullary bone and further demonstrate that this pterosaur reached reproductive maturity before skeletal maturity. This study shows that pterosaurs laid eggs even smaller than previously thought and had a reproductive strategy more similar to basal reptiles than to birds. Whether pterosaurs were highly precocial or needed parental care is still open to debate.

Chemical investigation on the root of R. laevigata led the isolation and identification of two new flavonoids, (+)-catechin-8-acetic acid (1) and guibourtacacidine 4-methyl ether (2), one known flavonoid, guibourtacacidine (3), along with seven known triterpenoids, euscaphic acid (4), kajiichigoside F1 (5), nigaichigoside F2 (6), rubuside J (7), tomentic acid (8), rosamutin (9) and betulinic acid (10). Their structures were established on the basis of spectroscopic evidence and comparisons with literature data. Compounds 1, 2 and 3 showed considerable radical scavenging activity in the 1, 1-diphenyl-2-picrylhydrazyl (DPPH) assay.

La investigación química de las raíces de R. laevigata condujo al aislamiento de dos nuevos flavonoides, 8-(+)-catequin-ácido acético (1) y éter 4-metílico de guibourtacacidina (2); un flavonoide conocido, guibourtacacidina (3), y siete triterpenos conocidos, ácido euscáfico (4), kajichi-gósido F1 (5), nigaichigósido F2 (6), rubúsido J (7), ácido toméntico (8), rosamutina (9) y ácido betulínico (10). Las estructuras fueron establecidas de acuerdo a las evidencias espectroscópicas y comparaciones con los datos informados en la literatura. Los compuestos 1, 2 y 3 mostraron actividad atrapadora de radicales en el ensayo del 1,1-difenil-2-picrilhidracilo (DFPH).

Purpose: The percutaneous renal access (PRA) is the most critical step of percutaneous renal surgery (PRS). For the training of PRA in the lab, a novel non-biological bench model was developed and set for validation test. Materials and Methods: Experts in PRS (> 60 cases) and novices were included to perform fluoroscopy guided PRA on the model. Overall time, X-ray exposure time and puncture attempts were recorded to establish construct validity. After accomplishment, the experts rated the model using a standardized questionnaire for face and content validity based on a 5-point Likert scale, with 1 denoting very bad and 5 as excellent. Baseline and post-training data of novices were analyzed for skill acquisition. Results: 9 experts and 30 novices were finally included. The overall appraisal was 4 by the experts, and consensus of all experts was reached for the model as an excellent training tool. Significant difference between experts and novices was detected with the experts using less total time 183.11 ± 29.40 vs. 278.00 ± 50.30 seconds (P < 0.001), shorter X-ray exposure time 109.22 ± 19.93 vs. 183.13 ± 38.83 seconds (P < 0.001), and fewer attempts 1.28 ± 0.44 vs. 2.35 ± 0.65 (P < 0.001). After training, the novices demonstrated significant skill improvement in total and fluoroscopy time, and number of attempts (P < 0.001). Conclusions: Our non-biological model provides a new method for PRA training. The face, content and construct validity were demonstrated. This model allows contact with PRA skills and could be applied to the first step in the learning curve.

The following study correlates three poems of Jose Watanabe (1946-2007) -"El Ojo", "Mi Casa", and "Como el Peje-sapo"- with the existentialist notion of the body. Three bodies are delimited, a biologic, conscious, and a cultural one, in order to facilitate the phenomenological description of doctor-patient relationship.

El estudio correlaciona tres poemas de José Watanabe (1946-2007) -"El ojo", "Mi casa" y "Como el peje-sapo"-con la noción existencialista de cuerpo. Se delimita tres cuerpos: el biológico, el consciente y el cultural, que facilitan la descripción fenomenológica de la relación médico- enfermo.