[Development and Application of Deep Learning-Based Model for Quality Control of Children Pelvic X-Ray Images].

Objective: A deep learning-based method for evaluating the quality of pediatric pelvic X-ray images is proposed to construct a diagnostic model and verify its clinical feasibility. Methods: Three thousand two hundred and forty-seven children with anteroposteric pelvic radiographs are retrospectively collected and randomly divided into training datasets, validation datasets and test datasets. Artificial intelligence model is conducted to evaluate the reliability of quality control model. Results: The diagnostic accuracy, area under ROC curve, sensitivity and specificity of the model are 99.4%, 0.993, 98.6% and 100.0%, respectively. The 95% consistency limit of the pelvic tilt index of the model is -0.052-0.072. The 95% consistency threshold of pelvic rotation index is -0.088-0.055. Conclusion: This is the first attempt to apply AI algorithm to the quality assessment of children's pelvic radiographs, and has significantly improved the diagnosis and treatment status of DDH in children.

Cation-exchange fibers and silver nanoparticles-modified carbon electrodes for selective removal of hardness ions and simultaneous deactivation of microorganisms in capacitive deionization.

The hardness and microorganism contamination are common problems of water quality around the world. Capacitive deionization (CDI) is a much-discussed solution to help solve the water crisis by providing efficient water softening while killing microorganism. Carboxylic (Na) cation-exchange fiber (CCEF) is an adsorbent material with good affinity for hardness ions. Silver nanoparticles (AgNPs) is a broad-spectrum microbicide. In this paper, the CCEF modified activated carbon (CCEF-AC) was used as cathode and showed excellent hardness ion adsorption selectivity at the optimum CCEF doping level (αCa2+/Na of 15.0, αMg2+/Na of 13.5). Its electrosorption capacity of Ca2+ reached 311 µmol/g, much higher than that of the AC cathode (188 µmol/g). It also showed good regenerable performance, retaining over 85 % of Ca2+ electrosorption capacity after 50 cycles stability test. The activated carbon modified with AgNPs (AC-Ag) was used as anode. When enhanced by an electric field, it could kill bacteria and microalgae with over 99 % and 90 % inhibition rates, respectively. This work has opened up a new way to simultaneously remove multiple pollutants (organic or inorganic) from water.

A novel approach for screening standard anteroposterior pelvic radiographs in children.

Anteroposterior pelvic radiography is the first-line imaging modality for diagnosing developmental dysplasia of the hip (DDH). Nonstandard radiographs with pelvic malposition make the correct diagnosis of DDH challenging. However, as the only method available for screening standard pelvic radiographs, traditional manual assessment is relatively laborious and potentially erroneous. We retrospectively collected 3,247 pelvic radiographs. There were 2,887 radiographs randomly selected to train and optimize the AI model. Then 362 radiographs were used to test the model's diagnostic performance. Its diagnostic accuracy was assessed using receiver operating characteristic (ROC) curves and measurement consistency using Bland-Altman plots. In 362 radiographs, the AI model's area under ROC curves, accuracy, sensitivity, and specificity for quality assessment was 0.993, 99.4% (360/362), 98.6% (138/140), and 100.0% (222/222), respectively. Compared with clinicians, the 95% limits of agreement (Bland-Altman analysis) for pelvic tilt index (PTI) and pelvic rotation index (PRI), as determined by the model, were -0.052-0.072 and -0.088-0.055, respectively. CONCLUSIONS: The artificial intelligence-assisted method was more efficient and highly consistent with clinical experts. This method can be used for real-time validation of the quality of pelvic radiographs in current picture archiving and communications systems (PACS). WHAT IS KNOWN: • Nonstandard pediatric radiographs with pelvic malposition make the correct diagnosis of developmental dysplasia of the hip (DDH) challenging. • Traditional manual assessment remains the only method available for screening standard pediatric pelvic radiographs, which is relatively laborious and potentially erroneous. WHAT IS NEW: • This study proposed an artificial intelligence-assisted model to assess the quality of pediatric pelvic radiographs accurately and efficiently. • We recommend the integration of the model into current picture archiving and communications systems (PACS) for real-time screening of standard pediatric pelvic radiographs.

Risk factors for misdiagnosis in children with developmental dysplasia of the hip: a retrospective single centre study.

OBJECTIVE: To investigate risk factors of misdiagnosis at the first visit of children with developmental dysplasia of the hip (DDH) who did not participate in hip ultrasound screening. METHODS: A retrospective review was conducted on children with DDH admitted to a tertiary hospital in northwestern China between January 2010 and June 2021. We divided the patients into the diagnosis and misdiagnosis groups according to whether they were diagnosed at the first visit. The basic information, treatment process and medical information of the children were investigated. We made a line chart of the annual misdiagnosis rate to observe the trend in the annual misdiagnosis rate. Univariate and multivariate logistic regression analyses were used to identify significant risk factors for missed diagnosis. RESULTS: A total of 351 patients met the inclusion criteria, including 256 (72.9%) patients in the diagnosis group and 95 (27.1%) patients in the misdiagnosis group. The line chart of the annual rate of misdiagnoses among children with DDH from 2010 to 2020 showed no significant change trend. Multiple logistic regression analysis showed that the paediatrics department (v the paediatric orthopaedics department: OR 0.21, p<0.001), the general orthopaedics department (v the paediatric orthopaedics department: OR 0.39, p=0.006) and the senior physician (v the junior physician: OR 2.47, p=0.006) on the misdiagnosis at the first visit of children were statistically significant. CONCLUSION: Children with DDH without hip ultrasound screening are prone to be misdiagnosed at their first visit. The annual misdiagnosis rate has not been significantly reduced in recent years. The department and title of the physician are independent risk factors for misdiagnosis.

Clinical thought-based software for diagnosing developmental dysplasia of the hip on pediatric pelvic radiographs.

Background: The common methods of radiographic diagnosis of developmental dysplasia of the hip (DDH) include measuring hip parameters and quantifying the degree of hip dislocation. However, clinical thought-based analysis of hip parameters may be a more effective way to achieve expert-like diagnoses of DDH. This study aims to develop a diagnostic strategy-based software for pediatric DDH and validate its clinical feasibility. Methods: In total, 543 anteroposterior pelvic radiographs were retrospectively collected from January 2017 to December 2021. Two independent clinicians measured four diagnostic indices to compare the diagnoses made by the software and conventional manual method. The diagnostic accuracy was evaluated using the receiver operator characteristic (ROC) curves and confusion matrix, and the consistency of parametric measurements was assessed using Bland-Altman plots. Results: In 543 cases (1,086 hips), the area under the curve, accuracy, sensitivity, and specificity of the software for diagnosing DDH were 0.988-0.994, 99.08%-99.72%, 98.07%-100.00%, and 99.59%, respectively. Compared with the expert panel, the Bland-Altman 95% limits of agreement for the acetabular index, as determined by the software, were -2.09°-2.91° (junior orthopedist) and -1.98°-2.72° (intermediate orthopedist). As for the lateral center-edge angle, the 95% limits were -3.68°-5.28° (junior orthopedist) and -2.94°-4.59° (intermediate orthopedist). Conclusions: The software can provide expert-like analysis of pelvic radiographs and obtain the radiographic diagnosis of pediatric DDH with great consistency and efficiency. Its initial success lays the groundwork for developing a full-intelligent comprehensive diagnostic system of DDH.

TLR2-hif1α-mediated glycolysis contributes to pyroptosis and oxidative stress in allergic airway inflammation.

As a pattern recognition receptor which activates innate immune system, toll-like receptor 2 (TLR2) has been reportedly mediates allergic airway inflammation (AAI), yet the underlying mechanism remains elusive. Here, in a murine AAI model, TLR2-/- mice showed decreased airway inflammation, pyroptosis and oxidative stress. RNA-sequencing revealed that allergen-induced hif1 signaling pathway and glycolysis were significantly downregulated when TLR2 was deficient, which were confirmed by lung protein immunoblots. Glycolysis inhibitor 2-Deoxy-d-glucose (2-DG) inhibited allergen-induced airway inflammation, pyroptosis, oxidative stress and glycolysis in wild type (WT) mice, while hif1α stabilizer ethyl 3,4-dihydroxybenzoate (EDHB) restored theses allergen-induced changes in TLR2-/- mice, indicating TLR2-hif1α-mediated glycolysis contributes to pyroptosis and oxidative stress in AAI. Moreover, upon allergen challenge, lung macrophages were highly activated in WT mice but were less activated in TLR2-/- mice, 2-DG replicated while EDHB reversed such effect of TLR2 deficiency on lung macrophages. Likewise, both in vivo and ex vivo WT alveolar macrophages (AMs) exhibited higher TLR2/hif1α expression, glycolysis and polarization activation in response to ovalbumin (OVA), which were all inhibited in TLR2-/- AMs, suggesting AMs activation and metabolic switch are dependent on TLR2. Finally, depletion of resident AMs in TLR2-/- mice abolished while transfer of TLR2-/- resident AMs to WT mice replicated the protective effect of TLR2 deficiency on AAI when administered before allergen challenge. Collectively, we suggested that loss of TLR2-hif1α-mediated glycolysis in resident AMs ameliorates allergic airway inflammation that inhibits pyroptosis and oxidative stress, therefore the TLR2-hif1α-glycolysis axis in resident AMs may be a novel therapeutic target for AAI.

Itaconate Suppresses the Activation of Mitochondrial NLRP3 Inflammasome and Oxidative Stress in Allergic Airway Inflammation.

Itaconate has emerged as a novel anti-inflammatory and antioxidative endogenous metabolite, yet its role in allergic airway inflammation (AAI) and the underlying mechanism remains elusive. Here, the itaconate level in the lung was assessed by High Performance Liquid Chromatography (HPLC), and the effects of the Irg1/itaconate pathway on AAI and alveolar macrophage (AM) immune responses were evaluated using an ovalbumin (OVA)-induced AAI model established by wild type (WT) and Irg1-/- mice, while the mechanism of this process was investigated by metabolomics analysis, mitochondrial/cytosolic protein fractionation and transmission electron microscopy in the lung tissues. The results demonstrated that the Irg1 mRNA/protein expression and itaconate production in the lung were significantly induced by OVA. Itaconate ameliorated while Irg1 deficiency augmented AAI, and this may be attributed to the fact that itaconate suppressed mitochondrial events such as NLRP3 inflammasome activation, oxidative stress and metabolic dysfunction. Furthermore, we identified that the Irg1/itaconate pathway impacted the NLRP3 inflammasome activation and oxidative stress in AMs. Collectively, our findings provide evidence for the first time, supporting the conclusion that in the allergic lung, the itaconate level is markedly increased, which directly regulates AMs' immune responses. We therefore propose that the Irg1/itaconate pathway in AMs is a potential anti-inflammatory and anti-oxidative therapeutic target for AAI.

Quantification of femoral position on radiographic acetabular coverage in children.

An accurate assessment of the radiographic acetabular coverage is essential for clinical diagnosis or surgical decision-making in hip disorders. This study aimed to evaluate the effect of femoral position on acetabular coverage and to predict the actual acetabular coverage from nonstandard radiographs. A total of 21 children (34 hips) with normative acetabular coverage were screened in this retrospective study. The Mimics-based local-rotation fluoroscopy simulation method was used to tilt, incline, and rotate the femur in 4° increments within the range of femoral motion. The acetabular coverage, namely acetabular-head index (AHI) and center-edge angle (CEA), increased with femoral abduction but decreased with other motions. Compared to the femoral neutral position, no significant differences were identified in AHI with the rotation (range: 0°-16°) and in CEA with the tilt (range: -20°-4°), inclination (range: 0°-4°), or rotation (range: -8°-40°). The linear regression analysis showed that the CEA increased by about 0.20° for each 1° increase in femoral inclination and decreased by about 0.01°, 0.07°, 0.06°, or 0.07° for each 1° increase in internal rotation, external rotation, flexion, or extension, respectively. And a more significant change in AHI was observed. All femoral malpositions, especially the inclination, affected radiographic acetabular coverage in children. Therefore, each pelvic radiograph should assess potential femoral malpositioning before diagnosing hip disorders. This study will assist surgeons in predicting the acetabular coverage on nonstandard radiographs.

Clinical reliability and validity of a video-based markerless gait evaluation method.

Objective: To explore the reliability and validity of gait parameters obtained from gait assessment system software employing a human posture estimation algorithm based on markerless videos of children walking in clinical practice. Methods: Eighteen typical developmental (TD) children and ten children with developmental dysplasia of the hip (DDH) were recruited to walk along a designated sidewalk at a comfortable walking speed. A 3-dimensional gait analysis (3D GA) and a 2-dimensional markerless (2D ML) gait evaluation system were used to extract the gait kinematics parameters twice at an interval of 2 h. Results: The two measurements of the children's kinematic gait parameters revealed no significant differences (P > 0.05). Intra-class correlation coefficients (ICC) were generally high (ICC >0.7), showing moderate to good relative reliability. The standard error of measurement (SEM) values of all gait parameters measured by the two walks were 1.26°-2.91°. The system software had good to excellent validity compared to the 3D GA, with ICC values between 0.835 and 0.957 and SEM values of 0.87°-1.71° for the gait parameters measured by both methods. The Bland-Altman plot analysis indicated no significant systematic errors. Conclusions: The feasibility of the markerless gait assessment method using the human posture estimation-based algorithm may provide reliable and valid gait analysis results for practical clinical applications.

Closing-opening wedge osteotomy for the treatment of congenital kyphosis in children.

BACKGROUND: To evaluate the safety and effectiveness of posterior closed-open wedge osteotomy for treatment of congenital kyphosis in children. METHODS: Imaging and clinical data from January 2010 to December 2019 of posterior closed-open wedge osteotomy of congenital kyphosis with at least 2-year follow up was analyzed retrospectively. Perioperative indicators such as operation time, osteotomy site, osteotomy method and occurrence of complications, and imaging indicators were observed. The 3D printed models were used to measure the expanded distance of anterior edge vertebra and closed length of spinal canal line. The clinical effect was evaluated through SRS-22 questionnaires. RESULTS: There were 15 CK patients in this study. The osteotomy segments and details are as follows: 1 case each for T6-9 and L2, 2 cases at T11, 3 cases at T12, and 6 cases at L1. The average operation time was 314 min, the average blood loss was 970 mL, the average fusion range was 6.3 segments, and the average time of follow up was 70.5 months. The Cobb angle of local kyphosis was corrected from 65.6 ± 18.8° to 11.3 ± 7.1°(p < .001). The range of kyphosis correction was 40-90°, and average correction rate was 83.2% (67.7-95.7%). The correction was stable in follow-up, and the kyphotic angle was 11.0 ± 7.6 (p = .68). The preoperative SVA was 31.5 ± 21.8 mm, and the postoperative recovery was 18.0 ± 15.5, while the last follow-up was 9.1 ± 7.9. The p values were 0.02 and 0.07 respectively. By using 3D printed models, the expanded distance of anterior edge vertebra and closed length of spinal canal line were 14.5 ± 7.5 mm and 24.5 ± 8.0 mm respectively. Self-image and satisfaction in SRS-22 improved significantly. There was no recurrence of deformity and junctional kyphosis. CONCLUSIONS: The posterior closing-opening wedge osteotom for treatment of congenital kyphosis in children is satisfactory, if selected appropriately. During the longitudinal follow-up, the patients could achieve solid fusion and the correction could be well maintained.Evidence of Confidence: IVa.

Comparison of Transfer Learning Models in Pelvic Tilt and Rotation Measurement in Pediatric Anteroposterior Pelvic Radiographs.

The rotation and tilt of the pelvis during anteroposterior pelvic radiography can lead to misdiagnosis of developmental dysplasia of the hip (DDH) in children. At present, no method exists for accurately and conveniently measuring the precise rotation and tilt angles of pelvic on radiographs. The objective of this study was to develop several rotation and tilt measurement models using transfer learning and digital reconstructed radiographs (DRRs), and to compare their performances on pelvic radiographs. Based on the inclusion criteria, 30 of 92 children who underwent 3D hip CT scans at Xijing Hospital from 2015 to 2020 were included in the study. Using DRR techniques, radiographs were generated by rotating and tilting the pelvis in CT datasets at - 12 to 12° (projected every 3°) and were randomized to a 2:1:1 ratio of training dataset, validation dataset, and test dataset. Five pre-trained networks, including VGG16, Xception, VGG19, ResNet50 and InceptionV3 were used to develop pelvic rotation measurement models and tilt measurement models, and these models were trained with training dataset. The callback function was used during the training to slow down the learning rate when learning was stalled. Then, the validation set was used to optimize each model and compare their performances. At last, we tested the final performances of optimal rotation measurement model and optimal tilt measurement model on test dataset. The mean absolute error (MAE) was employed to assess the performance of the models. A total of 2430 pelvic DRRs were collected based on 30 CT datasets. Among 5 pre-trained transfer learning models, VGG16-Tilt achieved the best tilt prediction performance at the same BS and different LR. VGG16-Tilt model achieved its best performance on validation set at LR = 0.001 and BS = 4, and the final MAE on the test set was 0.5250°. In terms of rotation prediction, VGG16-Rotation also achieved the best performance, and it achieved its best performance on validation set at LR = 0.002 and BS = 8. The final MAE of VGG16-Rotation on the test set was 1.0731°. Pretrained transfer learning models worked well in predicting tilt and rotation angles of the pelvis on radiographs in children. Among them, VGG16-Tilt and VGG16-Rotation had the best effect in dealing with such problems despite their simple structures. These models deployed in devices can give orthopedic surgeons a powerful aid in DDH diagnosis.

CpG Oligodeoxynucleotides Attenuate OVA-Induced Allergic Airway Inflammation via Suppressing JNK-Mediated Endoplasmic Reticulum Stress.

PURPOSE: CpG-ODN has been found to attenuate allergic airway inflammation in our previous study. Here, we aimed to further investigate whether CpG-ODN exerts such effect via regulating endoplasmic reticulum (ER) stress and revealed the underlying mechanism. METHODS: Five-week-old C57BL/6 mice were randomly grouped and treated with or without CpG-ODN or/and SP600125. Meantime, RAW264.7 cells were used to investigate the effect of CpG-ODN on OVA-induced ER stress in vitro. The cellularity of bronchoalveolar lavage fluid (BALF) was classified and counted after Wright-Giemsa staining. HE and PAS staining methods were applied to analyze airway inflammation. The protein levels of IL-4, IL-5, IL-13, p-JNK, JNK, CHOP, XBP1, ATF6α and GRP78 in lung tissues were detected by Western blotting. Correspondingly, the ER stress markers were detected by Western blotting and immunofluorescence in RAW264.7 cells. RESULTS: In OVA-induced allergic airway inflammation, CpG-ODN significantly suppressed inflammatory cells infiltration, goblet cell hyperplasia and the protein expression of Th2 cytokines. Moreover, OVA exposure strongly increased the activation of ER stress with higher protein expressions of CHOP, XBP1, ATF6α and GRP78. However, these OVA-induced increase of ER stress markers were markedly suppressed by CpG-ODN treatment. In addition, exposure to OVA significantly increased the phosphorylation of JNK, which was significantly reduced by CpG-ODN treatment. Remarkably, single treatment of SP600125, an antagonist of JNK, functioned similarly as CpG-ODN in mitigating allergic airway inflammation and suppressing OVA-induced activation of ER stress; however, no significant synergistic effect was evidenced by combined treatment of SP600125 and CpG-ODN. Furthermore, in OVA-stimulated RAW264.7 cells, we also found that OVA stimulation increased the expressions of ER stress markers, and CpG-ODN significantly reduced their expression levels via suppressing the phosphorylation of JNK. CONCLUSION: These results indicated that CpG-ODN mitigates allergic airway inflammation via suppressing the activation of JNK-medicated ER stress.

Inhibitory Effect and Potential Mechanism of Lactobacillus plantarum YE4 against Dipeptidyl Peptidase-4.

This study investigated the inhibitory effect and mechanism of 12 LAB strains isolated from Chinese fermented foods on dipeptidyl peptidase-4 (DPP-4) using the Caco-2 cell model. The results showed that the inhibitory effect of cell-free extracts (CFEs) collected from each LAB strain on DPP-4 was higher than that of the cell-free excretory supernatants. The CFEs from Lactobacillus plantarum YE4 (YE4-CFE) exhibited the strongest DPP-4 inhibitory activity (24.33% inhibition). Furthermore, YE4-CFE altered the TNF and MAPK signaling pathways. Additionally, the YE4-CFE ultrafiltration fraction (<3 kDa) displayed a similar DPP-4 inhibitory activity to YE4-CFE. UHPLC-MS/MS identified 19 compounds with a relative proportion of more than 1% in the <3 kDa fraction, and adenine, acetylcholine, and L-phenylalanine were the top three substances in terms of proportion. Altogether, the inhibitory effect of YE4-CFE on DPP-4 was associated with the TNF and MAPK signaling pathways, and with the high proportion of adenine, acetylcholine, and L-phenylalanine.

One-stage Surgical Treatment of Simultaneous Osteotomy and Asymmetric Lengthening on Short Femur with Severe Deformity of Genu Valgus.

This study aimed to discuss the effects of one stage surgical treatment by simultaneous osteotomy and asymmetric lengthening by Ilizarov external fixator on short femur with severe deformity of genu valgus. A total of 12 cases with unilateral deformities treated by simultaneous osteotomy and Ilizarov asymmetric lengthening on short femur with severe deformity of genu valgus were retrospectively analyzed from January 2006 to April 2015. The affected limbs were 2.5-11 cm (5.2 cm on average) short, the femorotibial angle was 135°-158° (146.3° on average), and the ankle interval was 15-43 cm (24.7 cm on average). The Paley method was used to determine the osteotomy plane (distal femur) of genu valgus. According to this standard, the bone union results were as follows: 11 had excellent and 1 had good, where 7 patients had excellent and 5 had good functional outcomes. One stage surgical treatment by simultaneous osteotomy and asymmetric lengthening on short femur with severe deformity of genu valgus was considered to be an effective and reliable method with better osteotomy union, less trauma and fewer complications.

Deficiency of Mkrn2 causes abnormal spermiogenesis and spermiation, and impairs male fertility.

Although recent studies have shed insights on some of the potential causes of male infertility, new underlining molecular mechanisms still remain to be elucidated. Makorin-2 (Mkrn2) is an evolutionarily conserved gene whose biological functions are not fully known. We developed an Mrkn2 knockout mouse model to study the role of this gene, and found that deletion of Mkrn2 in mice led to male infertility. Mkrn2 knockout mice produced abnormal sperms characterized by low number, poor motility, and aberrant morphology. Disruption of Mkrn2 also caused failure of sperm release (spermiation failure) and misarrangement of ectoplasmic specialization (ES) in testes, thus impairing spermiogenesis and spermiation. To understand the molecular mechanism, we found that expression of Odf2, a vital protein in spermatogenesis, was significantly decreased. In addition, we found that expression levels of Odf2 were decreased in Mkrn2 knockout mice. We also found that MKRN2 was prominently expressed in the sperm of normal men, but was significantly reduced in infertile men. This result indicates that our finding is clinically relevant. The results of our study provided insights into a new mechanism of male infertility caused by the MKRN2 downregulation.

Distinct temporal requirements for autophagy and the proteasome in yeast meiosis.

Meiosis is a special type of cellular renovation that involves 2 successive cell divisions and a single round of DNA replication. Two major degradation systems, the autophagy-lysosome and the ubiquitin-proteasome, are involved in meiosis, but their roles have yet to be elucidated. Here we show that autophagy mainly affects the initiation of meiosis but not the nuclear division. Autophagy works not only by serving as a dynamic recycling system but also by eliminating some negative meiotic regulators such as Ego4 (Ynr034w-a). In a quantitative proteomics study, the proteasome was found to be significantly upregulated during meiotic divisions. We found that proteasomal activity is essential to the 2 successive meiotic nuclear divisions but not for the initiation of meiosis. Our study defines the roles of autophagy and the proteasome in meiosis: Autophagy mainly affects the initiation of meiosis, whereas the proteasome mainly affects the 2 successive meiotic divisions.

Iqcg is essential for sperm flagellum formation in mice.

Mammalian spermatogenesis comprises three successive phases: mitosis phase, meiosis phase, and spermiogenesis. During spermiogenesis, round spermatid undergoes dramatic morphogenesis to give rise to mature spermatozoon, including the condensation and elongation of nucleus, development of acrosome, formation of flagellum, and removal of excessive cytoplasm. Although these transformations are well defined at the morphological level, the mechanisms underlying these intricate processes are largely unknown. Here, we report that Iqcg, which was previously characterized to be involved in a chromosome translocation of human leukemia, is highly expressed in the spermatogenesis of mice and localized to the manchette in developing spermatids. Iqcg knockout causes male infertility, due to severe defects of spermiogenesis and resultant total immobility of spermatozoa. The axoneme in the Iqcg knockout sperm flagellum is disorganized and hardly any typical ("9+2") pattern of microtubule arrangement could be found in Iqcg knockout spermatids. Iqcg interacts with calmodulin in a calcium dependent manner in the testis, suggesting that Iqcg may play a role through calcium signaling. Furthermore, cilia structures in the trachea and oviduct, as well as histological appearances of other major tissues, remain unchanged in the Iqcg knockout mice, suggesting that Iqcg is specifically required for spermiogenesis in mammals. These results might also provide new insights into the genetic causes of human infertility.

Altered protein prenylation in Sertoli cells is associated with adult infertility resulting from childhood mumps infection.

Mumps commonly affects children 5-9 yr of age, and can lead to permanent adult sterility in certain cases. However, the etiology of this long-term effect remains unclear. Mumps infection results in progressive degeneration of the seminiferous epithelium and, occasionally, Sertoli cell-only syndrome. Thus, the remaining Sertoli cells may be critical to spermatogenesis recovery after orchitis healing. Here, we report that the protein farnesylation/geranylgeranylation balance is critical for patients' fertility. The expression of geranylgeranyl diphosphate synthase 1 (GGPPS) was decreased due to elevated promoter methylation in the testes of infertile patients with mumps infection history. When we deleted GGPPS in mouse Sertoli cells, these cells remained intact, whereas the adjacent spermatogonia significantly decreased after the fifth postnatal day. The proinflammatory MAPK and NF-κB signaling pathways were constitutively activated in GGPPS(-/-) Sertoli cells due to the enhanced farnesylation of H-Ras. GGPPS(-/-) Sertoli cells secreted an array of cytokines to stimulate spermatogonia apoptosis, and chemokines to induce macrophage invasion into the seminiferous tubules. Invaded macrophages further blocked spermatogonia development, resulting in a long-term effect through to adulthood. Notably, this defect could be rescued by GGPP administration in EMCV-challenged mice. Our results suggest a novel mechanism by which mumps infection during childhood results in adult sterility.

Integrity of the blood-testis barrier in healthy men after suppression of spermatogenesis with testosterone and levonorgestrel.

STUDY QUESTION: Do exogenous male hormonal contraceptives that suppress intratesticular testosterone and spermatogenesis interfere with the blood-testis barrier integrity in men? SUMMARY ANSWER: When spermatogenesis was suppressed by testosterone alone or combined with levonorgestrel (LNG) treatment in men, the structural appearance of Sertoli cell tight junctions remained intact in the human testis. WHAT IS ALREADY KNOWN: Testosterone promotes the integrity of the blood-testis barrier. Intratesticular androgen deprivation induced by exogenous testosterone plus a progestin to suppress spermatogenesis in a contraceptive regimen may disturb the structural and functional integrity of the blood-testis barrier. STUDY DESIGN, SIZE AND DURATION: Testicular biopsies were obtained from a sub-study of a randomized clinical trial of 36 healthy Chinese men who were treated for 18 weeks and followed for at least a 12-week recovery period. PARTICIPANTS/MATERIAL, SETTING, METHODS: Healthy Chinese male volunteers (27-48 years) were randomized to two treatment groups (n = 18/group) for 18 weeks: (1) testosterone undecanoate (TU) 1000 mg i.m. injection followed by a 500 mg injection every 6 weeks and (2) TU + LNG 250 µg orally daily. Blood samples were obtained from all participants before and during treatment and at the end of the recovery phase. Open testicular biopsies for this study were obtained from four men before treatment and from four men in each of the TU and TU + LNG groups at 2 and 9 weeks of treatment. The presence of antisperm antibodies was checked in the archived serum samples of the subjects at baseline, during treatment and at the end of the recovery period. Stored testicular biopsy samples from cynomolgus monkeys treated with either sub-cutaneous testosterone or placebo for 12 weeks were used for additional protein expression studies. MAIN RESULTS AND ROLE OF THE CHANCE: Expression of blood-testis barrier associated proteins quantified by immunohistochemistry (claudin 3, claudin 11, junctional adhesion molecule-A, zonula occludens-1) remained unchanged despite a significant decrease in the numbers of pachytene spermatocytes and round spermatids in the seminiferous tubules at 9 weeks in the TU + LNG group. This was confirmed by immunoblots showing a lack of quantitative change in these tight junction proteins in monkeys after testosterone treatment. There were no increases in serum antisperm antibodies in the volunteers during the study. LIMITATIONS/REASONS FOR CAUTION: The duration of the study was short and the long-term effects of male hormonal contraceptive treatments on the integrity of the blood-testis barrier remain to be determined. WIDER IMPLICATIONS OF THE FINDINGS: This study supports the safety of male hormonal contraceptive treatment and does not corroborate the previous findings of disturbed immunological integrity of the blood-testis barrier from animal studies such as androgen receptor knockout mice and exogenous hormonal treatment in rats. STUDY FUNDING/COMPETING INTEREST: The study was supported by grants from the Contraceptive Research and Development Program and the Mellon Foundation (MFG-02-64, MFG-03-67), Endocrine, Metabolism and Nutrition Training Grant (T32 DK007571), the Clinical and Translational Science Institute at Los Angeles Biomedical and Harbor-UCLA Medical Center (UL1RR033176 and UL1TR000124) and the Los Angeles Biomedical Research Institute Summer High School Student Program.