Normal Morphology Research Articles

Trajectory model to analyze the effect of multi-metal exposures on sperm parameters and sex hormones of the MARHCS cohort in China

BackgroundAt present, several cross-sectional studies have found that exposure to metal/metalloid elements is closely associated with male reproduction. However, the long-term effects of metal exposure on male reproduction have not been explored. MethodsIn 2013, 796 volunteers were recruited, followed by first and second follow-ups in 2014 and 2015. Urine, semen, and blood samples were collected at each stage to examine urinary metal/metalloid levels, sperm parameters, and sex hormones. Initially, the latent class trajectory model (LCTM) was utilized to analyze the trajectories of urinary metals. Subsequently, the effects of urinary metal trajectories on semen parameters and sex hormones were examined using the linear mixed model. Finally, the impact of urinary metal trajectories on the classification of semen quality (normal or abnormal) was evaluated using the generalized linear mixed model. ResultsAmong the 18 metals/metalloids studied, trajectories were formed by 6 of them (Li, Al, Fe, Zn, As, Rb). Further analysis using the linear mixed model and the generalized linear mixed model revealed that Li was negatively correlated with semen volume, and sperm motility (P < 0.05). The maximum-decreasing trajectory group had a detrimental effect on semen quality (OR = 1.75, 95%CI: 1.22, 2.53) compared to the minimum-stable trajectory group. Al showed negative associations with sperm concentration, total sperm count, and normal morphology (P < 0.05). Rb was positively associated with progressive motility (P < 0.05). The high-stable trajectory group exhibited a protective effect on semen quality (OR = 0.66, 95%CI: 0.49, 0.90) compared to the low-stable trajectory group. Additionally, Fe was observed to have a negative relationship with follicle-stimulating hormone (FSH) (P < 0.05), and Rb exhibited a negative correlation with progesterone (P) (P < 0.05). ConclusionOur three-year cohort study provides new evidence that Li and Al have a negative impact on semen quality, whereas Rb is associated with beneficial effects. Additionally, Rb and Fe are endocrine disruptors of sex hormones.

Functional analysis of the whole CYPome and Fdxome of Streptomyces venezuelae ATCC 15439

Cytochrome P450 enzymes (CYPs or P450s) and ferredoxins (Fdxs) are ubiquitously distributed in all domains of life. Bacterial P450s are capable of catalyzing various oxidative reactions with two electrons usually donated by Fdxs. Particularly in Streptomyces, there are abundant P450s that have exhibited outstanding biosynthetic capacity of bioactive metabolites and great potential for xenobiotic metabolisms. However, no systematic study has been conducted on physiological functions of the whole cytochrome P450 complement (CYPome) and ferredoxin complement (Fdxome) of any Streptomyces strain to date, leaving a significant knowledge gap in microbial functional genomics. Herein, we functionally analyze the whole CYPome and Fdxome of Streptomyces venezuelae ATCC 15439 by investigating groups of single and sequential P450 deletion mutants, single P450 overexpression mutants, and Fdx gene deletion or repression mutants. Construction of an unprecedented P450-null mutant strain indicates that none of P450 genes are essential for S. venezuelae in maintaining its survival and normal morphology. The non-housekeeping Fdx1 and housekeeping Fdx3 not only jointly support the cellular activity of the prototypic P450 enzyme PikC, but also play significant regulatory functions. These findings significantly advance the understandings of the native functionality of P450s and Fdxs as well as their cellular interactions.

Influence of Cochlear Anatomy on Intraoperative Electrically Evoked Compound Action Potentials.

Objective: The electrically evoked compound action potential (ECAP) is an objective measure to indirectly assess spiral ganglion neurons. The ECAP provides inputs about the prognoses of cochlear implant (CI) recipients. Several factors such as cochlear morphology can affect ECAP measurements. This study aims to investigate the variation effect of cochlear parameters on intraoperative ECAP thresholds. Methods: This is a retrospective study on patients who underwent CI surgery with normal inner ear morphology at our center between 2017 and 2023. Cochlear anatomical parameters, including diameter (A value), width (B value), and height (H value), as well as cochlear duct length (CDL), were measured pre-operatively using OTOPLAN software (Version 3.0). Cochlear implant intraoperative objective measures were also collected. The correlation between the cochlear parameters and intraoperative objective measures was studied. Results: A total of 45 patients underwent cochlear implantation. The mean age was 2.4 ± 0.9 years. The mean CDL and cochlear coverage values were 33.2 ± 2.0 mm and 76.0 ± 5.7%, respectively. The ECAP threshold increased toward basal electrodes, with ECAP values as follows: apical 13.1 ± 3.8; middle 14.3 ± 3.7; and basal 15.6 ± 4.8. Additionally, the A, B, and H values showed a positive correlation with ECAP thresholds in different cochlear regions. The B value showed a significant moderate correlation with ECAP thresholds in the middle and basal electrodes but not in the apical electrodes. Conclusions: Cochlear anatomical parameters correlate with intraoperative ECAP thresholds. The B value showed a significant association with ECAP thresholds in the middle and basal electrodes. These findings could delineate the impact of the B value in CI and optimize electrode selection. Further research is required to study this correlation and its impact on postoperative outcomes.

Evaluation of sperm DNA fragmentation index among infertile individuals: a comprehensive analysis of the associated factors—a cross-sectional study

BackgroundEnsuring the integrity of sperm DNA is of paramount importance for the production of healthy offspring. The process of sperm formation involves intricate molecular adjustments that condense and safeguard its genetic material. However, the risk posed by sperm DNA fragmentation (SDF) can impede natural and assisted fertility and may arise from various testicular and post-testicular factors. While there are recommendations against routine SDF testing, recent research has sparked debate by demonstrating the positive impact of such testing on fertility outcomes.ObjectiveTo identify the factors associated with SDF among infertile individuals.Methods and materialsThe medical records of infertile men referred to our hospital were comprehensively reviewed and included in the study as they met the inclusion criteria. Patients dissatisfied with participation in the study or those with incomplete medical records were excluded. The association of SDF, sperm analysis, and demographic characteristics were evaluated for further investigation.ResultsThe average age of participants was 36.1 ± 8.1 years. The average semen volume was 2.5 ± 0.3 cc with a semen pH of 7.4 ± 0.2. The average sperm count was 10.9 ± 0.7 million, sperm motility was 36.9 ± 2.4%, and normal sperm morphology was 41.7 ± 2.7%. SDF levels below 15% were seen in 27.9%, between 15 and 30% in 32.4%, and above 30% in 39.7% of patients. Significant associations were found between the SDF and various factors in sperm analysis, including sperm count (P < 0.001), semen volume (P < 0.001), semen pH (P < 0.001), sperm motility (P < 0.001), normal sperm morphology (P < 0.001), and age (P < 0.001). Moreover, alcohol consumption (P = 0.04), smoking (P = 0.01), and the presence of varicocele (P = 0.03) were significantly associated with.ConclusionThis study emphasizes the significance of SDF among individuals experiencing infertility and investigates its correlation with various sperm test parameters. It also sheds light on the vital roles played by factors such as age, smoking, alcohol consumption, and the presence of varicocele, which are highly likely to cause variations in SDF levels. The results of our research will add to the existing pool of knowledge on infertility and potentially have implications for clinical practice and the care of patients.

Semen Parameters Can Be Used As A Credible Marker with Doppler Ultrasonography in the Diagnosis of Subclinical Varicocele Cases

OBJECTIVES: The link between varicoceles and male infertility has been a problem of debate for more than half a century. A substantial amount of data about varicocelectomies’ effects has been provided, but inadequate study designs and heterogeneity of current studies make these data rarely conclusive. This article investigates whether semen and sperm analysis in subclinical varicocele patients without any clinical signs diagnosed by Doppler Ultrasonography (USG) can be used as a diagnostic tool. STUDY DESIGN: The current prospective study included infertile male patients (n=44) enrolled in the Urology Clinic of Biruni University Hospital from January 2017 to January 2018. Patients were divided into two groups: 1st group as the control group (CG) (n=22): No varicocele as determined by Doppler USG, and 2nd group as the test group (TG) (n=22): Subclinical varicocele (SCV) as determined by Doppler USG. A semen analysis was done. Sperm were dyed with aniline blue and chromomycin A3 (CMA3), acridine orange (AO), propidium iodide (PI), and Rhodamine 123 (Rh123) for the determination of sperm maturation, sperm DNA fragmentation, apoptosis, and sperm mitochondrial membrane potential, respectively. Student-t Test was used for statistical analysis. p&lt;0.05 was considered significant. RESULTS: The sperm concentration of the CG was higher than the TG (p=0.98, p&gt;0.05). Forward sperm motility in the CG was higher than in the TG (p&lt;0.001, p&lt;0.05). Sperm with normal morphology in the TG was lower than in the CG (p&lt;0.001, p&lt;0.05). Sperm neck anomalies were higher in varicocele cases (p&lt;0.001, p&lt;0.05). Sperm maturation at the TG was lower than the CG (p&lt;0.001, p&lt;0.05). A high apoptotic sperm rate was found at the TG (p&lt;0.001, p&lt;0.05). Sperm mitochondria potential at the TG was lower than the CG (p&lt;0.001, p&lt;0.05). Sperm chromatin condensation at the TG was higher than the CG (p&lt;0.001, p&lt;0.05). CONCLUSIONS: The analysis of sperm DNA and apoptosis in SCV cases can be used as a reliable diagnostic tool to confirm Doppler USG.

Effect of dual tobacco smoking of hookah and cigarettes on semen parameters of infertile men.

The research regarding the effect of hookah smoking on health is still deficient, even though it has been proven to jeopardize human health by raising the hazard of different types of cancers, infections, and cardiovascular disease. We aimed to study the effect of dual tobacco smoking (hookah and cigarettes) on semen parameters of infertile men. In this cross-sectional study, we studied the effect of different types of smoking patterns on human semen parameters among men who visited IVF laboratories to do a seminal fluid analysis (SFA). A total number of 761 participants were included, divided into the following: 108 dual smokers, 219 hookah smokers, 222 cigarette smokers, and 212 non-smokers. To analyze the effect of dual smoking on normal morphology, an interaction term between the cigarette index and hookah index was used. Multivariable regression analysis after adjustment for age, BMI, education level, children, chronic diseases, varicocele, testicular surgery history, infertility duration, and cause revealed no significant difference in the sperm concentration and the percentage of progressive motility between non-smokers, cigarette smokers, or hookah smokers. However, there was a significant difference in the log of normal morphology percentage between the three groups. Cigarette and hookah smoking were significantly associated with having lower percentages of normal morphology. There was a significant difference in the log-normal morphology %, where light and heavy dual smokers had the least exponential beta of log-normal morphology %, 0.43 (95% CI: 0.33-0.55) and 0.36 (95% CI: 0.24-0.53), respectively. Dual tobacco smoking can adversely affect sperm morphology.

The potential protective effect of propolis on diabetic nephropathy induced by streptozotocin in adult albino rats

ABSTRACT Diabetes mellitus is a common metabolic disorder. It is associated with serious life-threatening complications if not properly managed. The current study aimed at investigating the possible protective role of propolis on streptozotocin-induced diabetic nephropathy. A diabetic rat model was induced by a single intraperitoneal injection of 55 mg/kg streptozotocin. After 4 days, the diabetic rats received oral propolis (300 mg/kg/day) via gastric gavage for 28 days. Biochemical, histopathological and ultrastructural evaluations were performed. The results showed that: streptozotocin-induced diabetes was associated with a marked decrease in the serum high-density lipoproteins and antioxidant enzymes. However, a significant elevation in the levels of serum creatinine, blood urea nitrogen, uric acid, cholesterol, triglycerides and low-density lipoproteins was detected. Furthermore, streptozotocin treatment induced histopathological alterations of the renal cortex; in the form of distorted glomerular capillaries, widened Bowman’s space and signs of epithelial tubular degeneration. Ultra-structurally, thickening and irregularity of the glomerular basement membrane and podocytes foot processes effacement were observed. The tubular epithelial cells showed swollen vacuolated mitochondria, scarce basal infoldings and loss of microvilli. Conversely, propolis partially restored the normal lipid profile, antioxidant biomarkers and renal cortical morphology. Propolis exhibited a sort of renoprotection through hypoglycemic, anti-hyperlipidemic and antioxidant effects.

Variations of the extrapsoas course of the lumbar plexus with implications for the lateral transpsoas approach to the lumbar spine: a cadaveric study

BackgroundTogether with an increased interest in minimally invasive lateral transpsoas approach to the lumbar spine goes a demand for detailed anatomical descriptions of the lumbar plexus. Although definitions of safe zones and essential descriptions of topographical anatomy have been presented in several studies, the existing literature expects standard appearance of the neural structures. Therefore, the aim of this study was to investigate the variability of the extrapsoas portion of the lumbar plexus in regard to the lateral transpsoas approach.MethodsA total of 260 lumbar regions from embalmed cadavers were utilized in this study. The specimens were dissected as per protocol and all nerves from the lumbar plexus were morphologically evaluated.ResultsThe most common variation of the iliohypogastric and ilioinguinal nerves was fusion of these two nerves (9.6%). Nearly in the half of the cases (48.1%) the genitofemoral nerve left the psoas major muscle already divided into the femoral and genital branches. The lateral femoral cutaneous nerve was the least variable one as it resembled its normal morphology in 95.0% of cases. Regarding the variant origins of the femoral nerve, there was a low formation outside the psoas major muscle in 3.8% of cases. The obturator nerve was not variable at its emergence point but frequently branched (40.4%) before entering the obturator canal. In addition to the proper femoral and obturator nerves, accessory nerves were present in 12.3% and 9.2% of cases, respectively.ConclusionNerves of the lumbar plexus frequently show atypical anatomy outside the psoas major muscle. The presented study provides a compendious information source of the possibly encountered neural variations during retroperitoneal access to different segments of the lumbar spine.

The effects of olibanum on male reproductive system damage in a lipopolysaccharide induced systemic inflammation model in rat

BackgroundLipopolysaccharide (LPS) as a particle of Gram-negative bacteria is a main contributer in the pathogenesis of the male reproductive system infectious. Male infertility due to LPS is reported to be related to overproduction reactive oxygen species. This study aimed to investigate the effects of olibanum on oxidative stress and apoptosis in testes and sperm dysfunction induced by LPS. MethodsThe male (n = 28) rats were allocated in four groups: control, LPS (1 mg/kg, i.p., 14 days), LPS + Olibanum 100 (100 mg/kg, i.p., 14 days), and LPS + Olibanum 200 (200 mg/kg, i.p., 14 days). Germ cell apoptosis was determined by TUNEL assays and computed using the stereological method. Additionally, semen samples of the animals were analyzed for sperm count and morphology. Oxidative stress indicators were also determined. ResultsThe count of TUNEL-positive germ cells in LPS-treated rats was more than that in the controls. Treatment of the animals with olibanum significantly attenuated the number of apoptotic cells compared to the LPS group. The sperm count and those with a normal morphology in LPS-treated rats was lower than that in the controls. Administration of olibanum significantly improved the sperms with normal morphology and sperm count. Olibanum treatment also improved superoxide dismutase, catalase, and total thiol in testicular tissue and decreased malondialdehyde. ConclusionAdministering both doses of olibanum in LPS-treated rats had potentially a therapeutic value in reducing germ cell apoptosis, as well as improving sperm parameters.

Mitochondrial dynamics regulate cell morphology in the developing cochlea.

In multicellular tissues, the size and shape of cells are intricately linked with their physiological functions. In the vertebrate auditory organ, the neurosensory epithelium develops as a mosaic of sensory hair cells (HCs), and their glial-like supporting cells, which have distinct morphologies and functional properties at different frequency positions along its tonotopic long axis. In the chick cochlea, the basilar papilla (BP), proximal (high-frequency) HCs, are larger than their distal (low-frequency) counterparts, a morphological feature essential for sound perception. Mitochondrial dynamics, which constitute the equilibrium between fusion and fission, regulate differentiation and functional refinement across a variety of cell types. We investigate this as a potential mechanism for regulating the shape of developing HCs. Using live imaging in intact BP explants, we identify distinct remodelling of mitochondrial networks in proximal compared with distal HCs. Manipulating mitochondrial dynamics in developing HCs alters their normal morphology along the proximal-distal (tonotopic) axis. Inhibition of the mitochondrial fusion machinery decreased proximal HC surface area, whereas promotion of fusion increased the distal HC surface area. We identify mitochondrial dynamics as a key regulator of HC morphology in developing inner ear epithelia.

N-Acetylcysteine Supplementation Improves Testicular Haemodynamics, Testosterone Levels, Seminal Antioxidant Capacity and Semen Quality in Heat-Stressed Goat Bucks.

Heat stress (HS) disrupts testicular homeostasis because of oxidative stress. N-acetylcysteine (NAC) is a thiol compound with antioxidants, anti-inflammatory and anti-apoptotic properties. As a sequel, this research aimed to assess the ameliorative effects of NAC supplementation on the reproductive performance of goat bucks kept under environmental HS. Primarily, Doppler examination as well as semen collection and evaluation were conducted on 12 mature bucks for 2 weeks (W) as pre-heat stress control (W1 and W2) during winter (February 2023). The temperature-humidity index (THI) was 63.4-64.3 (winter season). Then during summer HS conditions (from the beginning of July till the end of August 2023) bucks were assessed before NAC supplementation (W0), afterwards they were arbitrarily assigned into two groups. The control group (CON; n = 6) received the basal diet while the NAC group (n = 6) received the basal diet in addition to oral NAC daily for 7 weeks (W1-W7). The THI was 78.1-81.6 (summer season). Testicular blood flow parameters, serum concentration of nitric oxide (NO) and testosterone were measured. Additionally, total antioxidant capacity (TAC) and malondialdehyde (MDA) content in seminal plasma and semen quality parameters were evaluated. There were marked reductions (p < 0.05) in the resistive index (RI; W1, W4 and W5), pulsatility index (PI; W2 and W4-W7), and systolic/diastolic ratio (S/D; W4-W7) in the NAC group compared to the CON group. Furthermore, testosterone and NO levels were higher (p < 0.01 and p < 0.05, respectively) in the NAC group (W2, W3, W5 and W3-W5, respectively). Seminal plasma TAC increased (p < 0.05) and MDA decreased (p < 0.05) in the NAC group (W2, W4 and W5) compared to the CON group. Moreover, there were marked improvements (p < 0.05) in semen quality parameters (mass motility, total motility, viability and normal morphology) in the NAC group. In conclusion, oral NAC supplementation could be used to enhance the reproductive performance of goat bucks during HS conditions which is supported by remarkable enhancement in testicular haemodynamics, NO, testosterone levels and semen quality parameters.

The effectiveness and safety of the anti gastroesophageal reflux system in the treatment of gastroesophageal reflux disease: a single center study of 23 cases

Objective: To evaluate the safety and efficacy of domestically produced magnetic sphincter augmentation (MSA) for gastroesophageal reflux disease. Method: This study is a prospective cohort study. Patients with typical heartburn and reflux symptoms (at least partial response to proton pump inhibitors), abnormal esophageal acid exposure and normal esophageal peristalsis were included, prospectively in the Department of Gastroesophageal Surgery, Rocket Force Characteristic Medical Center from June 2019 to September 2022. Patients with hiatal hernia >2 cm and severe esophagitis were excluded. The MSA was wrapped around the distal esophagus after esophageal hiatus repair by laparoscopy. A postoperative questionnaire survey was conducted to assess the relief of symptom, complications, the discontinuation rate of proton pump inhibitor, and surgical satisfaction. Gastroscopy, high-resolution esophageal pressure measurement, and pH value impedance monitoring were also reviewed. The pre- and postoperative rates were compared using the McNeinar χ2 test. Result: Currently, 23 patients with gastroesophageal reflux disease were enrolled and underwent MSA surgery. There were 20 males and 3 females, aged (M (IQR)) 48 (14) years (range: 25 to 64 years). All cases were successfully implanted with MSA. Subjective indicators were followed for 17 (18) months (range: 14 to 53 months), while objective indicators were followed for 17 (1) months (range: 12 to 23 months). The postoperative gastrointestinal and extraesophageal symptom scores showed a significant decrease compared to preoperative levels as follows: the degree of subjective relief of overall digestive symptoms was 90 (20)% (range:0~100%), the degree of subjective relief of overall respiratory symptoms was 100(10)% (range: 10%~100%), the overall satisfaction rate was 83% (19/23), the proton pump inhibitor discontinuation rate was 70% (16/23). The proportion of esophagitis has decreased from 44% (10/23) to 9% (2/23) (κ=0.169, P=0.039), The Hill grade of gastroesophageal valve morphology improved from 1 case of grade Ⅰ, 5 cases of grade Ⅱ, 10 cases of grade Ⅲ, and 7 cases of grade Ⅲ preoperative to 22, 1, 0, and 0 cases postoperative. The proportion of lower esophageal sphincter pressure below normal has decreased from 70% (16/23) to 35% (8/23) (κ=0.170, P=0.012). There were 21 patients who restored normal esophageal acid exposure. Eleven patients had mild long-term dysphagia, but it didn't affect their daily life. No postoperative device migration, erosion, or secondary surgical removal occurred. Conclusions: Laparoscopic implantation of the MSA device was safe and well tolerated. It can effectively control the symptoms of gastroesophageal reflux disease, reduce medication, restore normal cardia morphology and function, and esophageal acid exposure. The main postoperative complication was dysphagia, but it was relatively mild.

Biochemical indicators, quality and safety of domestic and wild pig slaughter products

Animal husbandry is one of the important branches of agricultural production, which plays an important role in the formation of food security of Ukraine and providing the population with important types of food. The correct methodical approach to the inspection of organs and tissues of slaughtered animals allows to avoid errors in the diagnosis of various diseases, in the sanitary assessment of slaughter products, to prevent meat losses and the release of food products that are dangerous for humans. Analyzing the requirements of the modern market and the priority directions for the development of trade relations between the countries of the European Union for our country, it should be noted that the responsibility for the export and import of products of animal origin and the control of the production of high-quality and safe products is entrusted to the specialist of the state veterinary service. Organoleptic analysis of slaughter products is an important component of inspection. In addition, it should be noted that determining the degree of freshness of meat for a long time was carried out only with the help of the organoleptic method. It has been proven that organoleptic indicators provide comprehensive information for determining the good quality of meat. The quality of meat or its nutritional value refers to the chemical composition of the fleshy part, which includes muscle, fat and connective tissue. The chemical composition, energy value, digestibility, taste, culinary and other properties of meat depend on the ratio of these tissues. The quality of meat is influenced by the breed of animals, sex, age, fatness, the nature of fattening, conditions of transportation and pre-slaughter aging, methods of stunning, degree of exsanguination, etc. The article presents materials related to the inspection of animal slaughter products in the Zhytomyr region (agro-food market in the city of Olevsk). The study was carried out as part of the research work of the department of normal and pathological morphology, hygiene and expertise “Monitoring the content of toxic substances in drinking water, feed and their impact on the body of cattle and the quality and safety of products”, state registration number – 0120U101318. The purpose of the study is to inspect the slaughter products of domestic and wild pigs in the agro-food market of the Zhytomyr region. The task of the work was to determine the organoleptic, physico-chemical and microbiological indicators of slaughter products of domestic and wild pigs. The testing laboratory of veterinary and sanitary examination in the market carries out significant work on the inspection of animal slaughter products, which ensures the release of good-quality products for the population. As a result of the inspection of the slaughter products of domestic and wild pigs, it was established that all meat samples according to organoleptic parameters corresponded to the parameters of fresh high-quality meat, the reaction to peroxidase was positive, the pH of the meat ranged from 5.9 to 6.1. No trichinella was detected during trichinoscopy of pork samples. Veterinary and sanitary examination of livestock products on the markets is carried out only by veterinary medicine specialists of state laboratories or other state institutions of veterinary medicine. The laboratory carries out mandatory quality and safety control of all controlled products sold, including food products of animal origin intended for human consumption. In this regard, it is important to carry out inspection, sanitary evaluation of slaughter products of slaughter animals and determination of morphological characteristics of meat of domestic and wild pigs.

Skeletal Stem Cell–Derived Exosomes Promote Meniscal Tear Healing and Ameliorate Secondary Osteoarthritis

Background: The self-repair ability after meniscal tears is poor, leading to the development of posttraumatic osteoarthritis. Promoting the repair of meniscal injuries remains a great challenge, especially in the avascular region. Hypothesis: Local delivery of skeletal stem cell (SSC)–derived exosomes (SSC-Exos) would promote meniscal healing and prevent secondary osteoarthritis progression. Study Design: Controlled laboratory study. Methods: SSCs were isolated from bone marrow and exosomes were extracted via ultracentrifugation. The cell migration capabilities after incubation with exosomes were validated through in vitro cell culture. Full-thickness longitudinal medial meniscal tears were performed in the avascular region of 40 male Sprague-Dawley rats and 20 male New Zealand White rabbits, which were randomly divided into 2 groups: group treated with phosphate-buffered saline (GCON) and group treated with exosomes (GExosome). The effects of these treatments on meniscal healing and secondary osteoarthritis were evaluated by gross inspection, biomechanical testing, and histological assessment. RNA sequencing of in vitro cell cultures was performed to explore the underlying mechanisms. Results: Exosomes were successfully extracted and identified. These exosomes significantly promoted cell migration capabilities in vitro (P < .01). The GExosome exhibited greater cell proliferation and tissue regeneration with type 2 collagen secretion, and a significantly higher meniscal repair score than that of the GCON at 8 weeks postoperatively (P < .05). In contrast to the degenerative changes in both the meniscus and articular cartilage of the GCON, meniscal tissue in the GExosome exhibited restoration of normal morphology with a smooth and glossy white surface and better mechanical strength at 8 weeks after meniscal repair. Both degeneration scores and synovitis scores were significantly higher in the GCON than in the GExosome (P < .05). Compared with the GCON, the expression of key genes related to cell migration, such as the chemokine family, was enhanced by exosome injection, leading to an upregulation of extracellular matrix expression while downregulating the expression of inflammation-related genes such as CD68 and the matrix metalloproteinase family. Conclusion: The administration of SSC-Exos effectively promoted meniscal healing in the avascular region and ameliorated secondary osteoarthritis. The effect might be attributed to inflammation modulation, promotion of cell migration, and secretion of extracellular matrix components. Clinical Relevance: Injection of SSC-Exos represents a promising therapeutic option for promoting meniscal healing in the avascular region.

Geometric Morphometric Study on Distinguishing Metopic Craniosynostosis from Metopic Ridging.

Craniosynostosis, a common congenital anomaly, results from premature fusion of the cranial sutures. One of the forms of craniosynostosis is premature fusion of the metopic suture, referred to as trigonocephaly, but the diagnosis of metopic suture synostosis remains controversial. The purpose of this study was to clarify, using geometric morphometric analysis, if a metopic ridge alone observed in cases of mild trigonocephaly represents a pathological phenomenon. Three different cranial morphologies were compared among patients up to 2 years old who were categorized into the true group, the mild group, and the normal group, based on the presence or absence of specific symptoms, history of cranioplasty for trigonocephaly, or lack of any abnormality on computed tomography. Using the obtained computed tomography images, 235 anatomical landmarks and semi-landmarks were plotted on the entire cranial surface for analysis of neurocranial morphology, and the cranial shapes represented by landmarks were analyzed using geometric morphometrics. Principal components of shape variations among specimens were then computed, based on the variance-covariance matrix of the Procrustes residuals of all specimens, and statistically analyzed. The principal component analyses of the variations in endocranial shape, frontal bone shape, and occipital bone shape did not show any significant differences in cranial morphology between mild trigonocephaly and normal skulls; however, true trigonocephaly was found to differ significantly from mild trigonocephaly and normal skulls. These findings suggest that in assessments of cranial morphology, the presence of a ridge alone cannot be diagnosed as fundamentally pathological, and may represent normal morphology.

Differentiating idiopathic inflammatory myopathies by automated morphometric analysis of MHC-1, MHC-2 and ICAM-1 in muscle tissue.

Diagnosis of idiopathic inflammatory myopathies (IIM) is based on morphological characteristics and the evaluation of disease-related proteins. However, although broadly applied, substantial bias is imposed by the respective methods, observers and individual staining approaches. We aimed to quantify the protein levels of major histocompatibility complex (MHC)-1, (MHC)-2 and intercellular adhesion molecule (ICAM)-1 using an automated morphometric method to mitigate bias. Double immunofluorescence staining was performed on whole muscle sections to study differences in protein expression in myofibre and endomysial vessels. We analysed all IIM subtypes including dermatomyositis (DM), anti-synthetase syndrome (ASyS), inclusion body myositis (IBM), immune-mediated-necrotising myopathy (IMNM), dysferlinopathy (DYSF), SARS-CoV-2 infection and vaccination-associated myopathy. Biopsies with neurogenic atrophy (NA) and normal morphology served as controls. Bulk RNA-Sequencing (RNA-Seq) was performed on a subset of samples. Our study highlights the significance of MHC-1, MHC-2 and ICAM-1 in diagnosing IIM subtypes and reveals distinct immunological profiles. RNASeq confirmed the precision of our method and identified specific gene pathways in the disease subtypes. Notably, ASyS, DM and SARS-CoV-2-associated myopathy showed increased ICAM-1 expression in the endomysial capillaries, indicating ICAM-1-associated vascular activation in these conditions. In addition, ICAM-1 showed high discrimination between different subgroups with high sensitivity and specificity. Automated morphometric analysis provides precise quantitative data on immune-associated proteins that can be integrated into our pathophysiological understanding of IIM. Further, ICAM-1 holds diagnostic value for the detection of IIM pathology.

Generation of a human SLC12A3 knock-in human induced pluripotent stem cell line (CMCi014-A-82) using CRISPR-Cas9 system

Gitelman’s disease is caused by a genetic mutation in the solute carrier family 12 member 3 (SLC12A3) gene, which encodes the sodium chloride cotransporter. In this study, we generated a stable human induced pluripotent stem cell (hiPSC) line, WTC11-SLC12A3 (CMCi014-A-82), by knocking in the entire SLC12A3 gene at the SHS231 locus in healthy wild-type control hiPSCs (WTC11). We verified that WTC11-SLC12A3 expressed pluripotency markers and exhibited normal stem cell morphology. Furthermore, this cell line maintains a normal karyotype and can differentiate into the three germ layers. Therefore, this cell line may provide a basis for gene therapy for Gitelman’s disease.

Magnetic resonance imaging pattern recognition of metabolic and neurodegenerative encephalopathies in dogs and cats.

Metabolic/neurodegenerative encephalopathies encompass a wide list of conditions that share similar clinical and magnetic resonance imaging (MRI) characteristics, challenging the diagnostic process and resulting in numerous tests performed in order to reach a definitive diagnosis. The aims of this multicentric, retrospective and descriptive study are: (I) to describe the MRI features of dogs and cats with metabolic/neurodegenerative encephalopathies; (II) to attempt an MRI recognition pattern classifying these conditions according to the involvement of grey matter, white matter or both; and (III) to correlate the MRI findings with previous literature. A total of 100 cases were recruited, comprising 81 dogs and 19 cats. These included hepatic encephalopathy (20 dogs and three cats), myelinolysis (five dogs), intoxications (seven dogs and one cat), thiamine deficiency (two dogs and seven cats), hypertensive encephalopathy (three dogs and two cats), neuronal ceroid lipofuscinosis (11 dogs and one cat), gangliosidosis (three dogs and two cats), fucosidosis (one dog), L-2-hydroxyglutaric aciduria (13 dogs and one cat), Lafora disease (11 dogs), spongiform leukoencephalomyelopathy (one dog) and cerebellar cortical degeneration (four dogs and two cats). None of the hepatic encephalopathies showed the previously described T1-weighted hyperintensity of the lentiform nuclei. Instead, there was involvement of the cerebellar nuclei (8/23), which is a feature not previously described. Dogs with myelinolysis showed novel involvement of a specific white matter structure, the superior longitudinal fasciculus (5/5). Thiamine deficiency affected numerous deep grey nuclei with novel involvement of the oculomotor nuclei (3/9), thalamic nuclei, subthalamus and cerebellar nuclei (1/9). Cats with hypertensive encephalopathy had a more extensive distribution of the white matter changes when compared to dogs, extending from the parietal and occipital lobes into the frontal lobes with associated mass effect and increased brain volume. Lysosomal storage disease showed white matter involvement only, with neuronal ceroid lipofuscinosis characterised by severe brain atrophy when compared to gangliosidosis and fucosidosis. All patients with L-2-hydroxyglutaric aciduria had a characteristic T2-weighted hyperintense swelling of the cerebral and cerebellar cortical grey matter, resulting in increased brain volume. Lafora disease cases showed either normal brain morphology (5/11) or mild brain atrophy (6/11). Dogs with cerebellar cortical degeneration had more marked cerebellar atrophy when compared to cats. This study shows the important role of MRI in distinguishing different metabolic/neurodegenerative encephalopathies according to specific imaging characteristics.

Pharmacological non-hormonal treatment options for male infertility: a systematic review and network meta-analysis

BackgroundMale factor infertility affect up to 50% of couples unable to conceive spontaneously. Several non-hormonal pharmacological treatments have been proposed to boost spermatogenesis and increase chances of conception in men with infertility. Still, no clear evidence exists on the most effective treatment strategy.ObjectiveWe aimed to compare the effectiveness of non-hormonal pharmacological treatment options for men with infertility using a systematic review and network meta-analysis.MethodsWe searched MEDLINE, EMBASE, and CENTRAL until October 2023 for randomised/quasi-randomised trials that evaluated any non-hormonal pharmacological treatment options for men with idiopathic semen abnormalities or those with hypogonadism. We performed pairwise and network meta-analyses using a random effect model. We assessed risk of bias, heterogeneity, and network inconsistency. We calculated the mean rank and the surface under the cumulative ranking curve (SUCRA) for each intervention the maximum likelihood to achieve each of reported outcomes. We reported primarily on sperm concentration and other important semen and biochemical outcomes using standardised mean difference (SMD) and 95% confidence-intervals(CI).ResultsWe included 14 randomised trials evaluating four treatments (Clomiphene citrate, Tamoxifen, Aromatase inhibitors, anti-oxidants) and their combinations in 1342 men. The overall quality of included trials was low. Sperm concentration improved with clomiphene compared to anti-oxidants (SMD 2.15, 95%CI 0.78–3.52), aromatase inhibitor (SMD 2.93, 95%CI 1.23–4.62), tamoxifen (SMD − 1.96, 95%CI -3.57; -0.36) but not compared to placebo (SMD − 1.53, 95%CI -3.52- 0.47). Clomiphene had the highest likelihood to achieve the maximum change in sperm concentration (SUCRA 97.4). All treatments showed similar effect for sperm motility, semen volume, and normal sperm morphology. FSH levels showed significant improvement with clomiphene vs.anti-oxidant (SMD 1.48, 95%CI 0.44–2.51) but not compared to placebo. The evidence networks for LH and testosterone suffered from significant inconsistency (p = 0.01) with similar trend of improvement with clomiphene compared to other treatments but not compared to placebo.ConclusionThere is insufficient evidence to support the routine use of Clomiphene, tamoxifen, and aromatase inhibitors to optimise semen parameters in men with infertility. Future randomised trials are needed to confirm the efficacy of clomiphene in improving fertility outcomes in men.PROSPEROCRD42023430179.

Pregnancy and perinatal outcomes in pregnancies following frozen embryo transfer (FET) after transcervical resection of adhesions (TCRA): A retrospective cohort study with propensity score matching analysis.

To compare the pregnancy and perinatal outcomes of frozen-thawed embryo transfer (FET) in patients following transcervical resection of adhesions (TCRA) versus patients with normal uterine morphology, and to investigate the factors influencing pregnancy outcomes in patients undergoing FET after TCRA. We retrospectively analyzed FET cycles from September 2014 to September 2023, comparing patients with normal uterine morphology to those with intrauterine adhesions (IUAs) treated with TCRA. Propensity score matching (PSM) adjusted for confounding factors. LASSO regression and multivariate logistic regression identified predictors of outcomes, which were visually represented in nomograms. Model performance was assessed using calibration curves, ROC curves, and DCA, with bootstrap method for internal validation. Post-PSM analysis showed higher live birth rates in patients with normal uterine morphology after clinical pregnancy (75.1% vs. 61.7%, P < 0.001). No significant differences were noted in clinical pregnancy rates and perinatal outcomes between the groups. Factors influencing clinical pregnancy in FET after TCRA included basal progesterone levels, endometrial thickness, parity, infertility cause, embryo stage at transfer, number and quality of embryos transferred, IUA severity, and TCRA surgical procedures. Body mass index, basal LH levels, and day 14 HCG levels post-embryo transfer were determinants of live birth outcome. FET cycles following TCRA showed a lower rate of successful live births, but TCRA did not increase adverse perinatal outcome risks. Our study introduces an innovative predictive model for clinical pregnancy and live birth outcomes in patients undergoing FET following TCRA, addressing a significant void in existing research.