ABSTRACT
The regulation of spermatogonial proliferation and apoptosis is of great significance for maintaining spermatogenesis. The single-cell RNA sequencing (scRNA-seq) analysis of the testis was performed to identify genes upregulated in spermatogonia. Using scRNA-seq analysis, we identified the spermatogonia upregulated gene origin recognition complex subunit 6 (Orc6), which is involved in DNA replication and cell cycle regulation; its protein expression in the human and mouse testis was detected by western blot and immunofluorescence. To explore the potential function of Orc6 in spermatogonia, the C18-4 cell line was transfected with control or Orc6 siRNA. Subsequently, 5-ethynyl-2-deoxyuridine (EdU) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays, flow cytometry, and western blot were used to evaluate its effects on proliferation and apoptosis. It was revealed that ORC6 could promote proliferation and inhibit apoptosis of C18-4 cells. Bulk RNA sequencing and bioinformatics analysis indicated that Orc6 was involved in the activation of wingless/integrated (Wnt)/ β-catenin signaling. Western blot revealed that the expression of β-catenin protein and its phosphorylation (Ser675) were significantly decreased when silencing the expression of ORC6. Our findings indicated that Orc6 was upregulated in spermatogonia, whereby it regulated proliferation and apoptosis by activating Wnt/β-catenin signaling.
ABSTRACT
Klinefelter syndrome (KS) is the most common genetic cause of human male infertility. However, the effect of the extra X chromosome on different testicular cell types remains poorly understood. Here, we profiled testicular single-cell transcriptomes from three KS patients and normal karyotype control individuals. Among the different somatic cells, Sertoli cells showed the greatest transcriptome changes in KS patients. Further analysis showed that X-inactive-specific transcript ( XIST ), a key factor that inactivates one X chromosome in female mammals, was widely expressed in each testicular somatic cell type but not in Sertoli cells. The loss of XIST in Sertoli cells leads to an increased level of X chromosome genes, and further disrupts their transcription pattern and cellular function. This phenomenon was not detected in other somatic cells such as Leydig cells and vascular endothelial cells. These results proposed a new mechanism to explain why testicular atrophy in KS patients is heterogeneous with loss of seminiferous tubules but interstitial hyperplasia. Our study provides a theoretical basis for subsequent research and related treatment of KS by identifying Sertoli cell-specific X chromosome inactivation failure.
Subject(s)
Animals , Humans , Male , Female , Sertoli Cells/metabolism , Klinefelter Syndrome/genetics , Endothelial Cells , Testis/metabolism , X Chromosome/metabolism , Mammals/geneticsABSTRACT
Non-obstructive azoospermia (NOA), which is defined as the absence of spermatozoa in the ejaculate secondary to impaired spermatogenesis within the testis, may be caused by a variety of etiologies, including varicocele-induced testicular damage, cryptorchidism, prior testicular torsion, post-pubertal mumps orchitis, gonadotoxic effects from medications, genetic abnormalities, chemotherapy/radiation, and other unknown causes currently classified as idiopathic (Cocuzza et al., 2013). The microdissection testicular sperm extraction (micro-TESE) technique involves a meticulous microsurgical exploration of the testicular parenchyma to identify and selectively extract larger seminiferous tubules that carry a higher probability of complete spermatogenesis (Schlegel, 1999). The Cornell group evaluated the efficacy of micro-TESE in 152 NOA patients with an associated history of cryptorchidism. In their series, spermatozoa were successfully retrieved in 116/181 attempts (64%), and the resulting pregnancy rate was 50% with a delivery rate of 38% (Dabaja and Schlegel, 2013). Franco et al. (2016) described a stepwise micro-TESE approach in NOA patients, which was considered to reduce the cost, time, and effort associated with the surgery. Alrabeeah et al. (2016) further reported that a mini-incision micro-TESE, carried through a 1-cm equatorial testicular incision, can be useful for micro-TESE candidates, particularly in patients with cryptozoospermia. We conducted a retrospective study of 20 consecutive NOA patients with a history of orchidopexy from May 2015 to March 2017.
Subject(s)
Adult , Humans , Male , Middle Aged , Azoospermia/surgery , Microdissection/methods , Orchiopexy , Retrospective Studies , Sperm RetrievalABSTRACT
We retrospectively reviewed data for 286 patients with varicocele who underwent microsurgical subinguinal varicocelectomy from March 2015 to May 2017 in Shanghai General Hospital (Shanghai, China). In this surgical approach, the testis was delivered, and the gubernacular and external cremasteric veins were stripped. In addition, the spermatic cord was delivered downward with continuous double traction away from the external ring. The remaining procedure was similar to the conventional approach. We followed patients for at least 3 months and evaluated postoperative semen parameters, pain symptoms, and complications. We excluded data for 32 men due to inadequate follow-up (<3 months). Of the remaining 254 patients, 73 had oligoasthenospermia, 121 had nonobstructive azoospermia, and 60 had symptomatic varicoceles. Total progressive sperm counts increased in the oligoasthenospermic patients from a median preoperative value of 9.15 × 106 ml-1 to 25.33 × 106 ml-1 (n= 34), and 35.6% (26/73) initially oligoasthenospermic men contributed to unassisted pregnancies. Sperm returned to the ejaculate in 12.4% (15/121) azoospermia patients. In patients with scrotal pain (n = 60), 43 (71.7%) reported complete resolution of pain, 16 (26.7%) reported partial resolution, and 1 (1.7%) reported no change. No patients experienced varicocele recurrence. This double-traction strategy avoids opening the external oblique aponeurosis, and results in less damage and faster recovery. In addition, the stripping strategy eliminates potential damage to the testis caused by the varicose veins. Our results showed that microsurgical subinguinal varicocelectomy using spermatic cord double traction in conjunction with testicular delivery for vein stripping is a safe and effective approach for varicocele repair.
Subject(s)
Adolescent , Adult , Humans , Male , Middle Aged , Young Adult , Microsurgery , Retrospective Studies , Spermatic Cord/surgery , Testis/blood supply , Treatment Outcome , Urogenital Surgical Procedures/methods , Varicocele/surgery , Veins/surgeryABSTRACT
In China, there are three basic clinical decision-making modes for patients, namely patients' autonomous decision-making mode, family decision-making mode and patient and family codetermination. They were produced under the unique background of Chinese medicine, Confucian philosophy and law in China. In this paper, the concepts, advantages and disadvantages of these three decision-making modes were analyzed. In addition, some suggestions were put forward for the improvement. The first is that we suggest to establish standards for choosing decision-making modes; the second is to further learn and publicize relevant laws; thirdly, the legal system needs to be further refined; and the last one is to carry out ethical ward round.
Subject(s)
Humans , China , Decision Making , Patient ParticipationABSTRACT
Objective: To observe the expression of osteopontin (OPN) in the giant cell tumor of bone (GCTB) and to discuss its clinical significance. Methods: OPN expression was examined in 77 GCTB samples by En Vision™ two-step immunohistochemical method. And the expression was compared between patients with different clinical and pathological data; the implied clinical relevance was also analyzed. Results: The positive expression rate of OPN in the GCTB was 76.6%; only weak expression was noted in the normal bone tissues. The expression in the invasive GCTB was significantly higher than that in the non-invasive ones(95.5% vs 69.1%, P=0.013). The expression rate of OPN increased with the increase of Campanacci stages of GCTB, and significant difference was found between different stages(P=O.019). The recurrence status, gender, age, surgery types, duration of illness or site of tumor were not associated with the positive rate of OPN. Conclusion: OPN is highly expressed in GCTB tissues, and the expression is associated with the invasiveness and the Campanacci stages of GCTB.
ABSTRACT
<p><b>OBJECTIVE</b>To study the clinical significance of osteopontin (OPN) and its receptor CD44v6 expression in oral squamous cell carcinoma (OSCC).</p><p><b>METHODS</b>OPN and CD44v6 expression were examined in OSCC (n=59) and normal oral mucosa (n=12) by EnVision method, the staining-grade were quantitatively studied by multiple functional image analyzer. Their expression grade of different clinical and pathological index were statistically studied.</p><p><b>RESULTS</b>OPN expression grade was significantly higher in OSCC than that in normal oral mucosa (P<0.05). Significant deviation of OPN expression grade was found between different clinical stages, as well as between the groups with or without cervical lymph node metastasis. The group with cervical lymphnode metastasis had higher expression than that of the group without lymph node metastasis (P<0.05). However, there was no significant deviation between the expression grade in well-differentiated group and moderate or poorly differentiated group. The expression of CD44v6 showed no correlation with that of OPN, nor any difference between OSCC and normal oral mucosa.</p><p><b>CONCLUSION</b>OPN over expression was found in OSCC, and the expression level has correlation with the clinical staging and with cervical lymph node metastasis status. CD44v6 expression showed no difference between OSCC and normal oral mucosa nor any correlation with that of OPN.</p>