Circular RNA circRSF1 binds to HuR to promote radiation-induced inflammatory phenotype in hepatic stellate cells / 南方医科大学学报

OBJECTIVE@#To investigate whether circular RNA circRSF1 regulates radiation-induced inflammatory phenotype of hepatic stellate cells (HSCs) by binding to HuR protein and repressing its function.@*METHODS@#Human HSC cell line LX2 with HuR overexpression or knockdown was exposed to 8 Gy X-ray irradiation, and the changes in the expression of inflammatory factors (IL-1β, IL-6 and TNF-α) were detected by qRT-PCR. The expressions of IκBα and phosphorylation of NF-κB were detected with Western blotting. The binding of circRSF1 to HuR was verified by RNA pull-down assay and RNA-binding protein immunoprecipitation (RIP). The expressions of inflammatory factors, IκBα and the phosphorylation of NF-κB were detected after modifying the interaction between circRSF1 and HuR.@*RESULTS@#Knockdown of HuR significantly up- regulated the expressions of IL-1β, IL-6 and TNF-α, decreased IκBα expression and promoted NF-κB phosphorylation in irradiated LX2 cells, whereas overexpression of HuR produced the opposite changes (P < 0.05). Overexpression or knockdown of circRSF1 did not significantly affect the expression of HuR. RNA pull-down and RIP experiments confirmed the binding between circRSF1 and HuR. Overexpression of circRSF1 significantly reduced the binding of HuR to IκBα and down-regulated the expression of IκBα (P < 0.05). Overexpression of circRSF1 combined with HuR overexpression partially reversed the up-regulation of the inflammatory factors, down-regulated IκBα expression and increased phosphorylation of NFκB in LX2 cells, while the opposite effects were observed in cells with knockdown of both circRSF1 and HuR (P < 0.05).@*CONCLUSION@#circRSF1 reduces IκBα expression by binding to HuR to promote the activation of NF-κB pathway, thereby enhancing radiation- induced inflammatory phenotype of HSCs.

Surgical management and molecular diagnosis of persistent Müllerian duct syndrome in Chinese patients / 亚洲男科学杂志(英文版)

Persistent Müllerian duct syndrome (PMDS) is a rare clinically and genetically overlapping disorder caused by mutations in the anti-Müllerian hormone (AMH) gene or the anti-Müllerian hormone receptor type 2 (AMHR2) gene. Affected individuals present uterus and tubes in normally virilized males and are discovered unexpectedly during other surgeries. Since it is rare and complex, a definitive clinical diagnosis can be missed, and there are no guidelines regarding how to deal with the uterus. In the present study, exome sequencing and Sanger verification were performed for causal variants in 12 PMDS patients. Preoperative diagnoses were made by positive exome sequencing in 8 patients. Of them, 7 patients evoked on the basis of ultrasound indicating bilateral testes on the same side of the body. Twelve different AMH variants (2 frameshift/nonsense, 1 deletion, 8 missense, and 1 in-frame) in 9 patients and 6 different AMHR2 variants (5 missense and 1 splicing) in 3 patients were identified. Seven variants were classified as "pathogenic" or "likely pathogenic", and 4 of them were novel. All but two patients with AMH defects showed low serum AMH concentrations, but all patients with AMHR2 defects showed elevated AMH levels. During surgery, an abnormal vas deferens was observed in half of the patients. Eight patients underwent orchidopexy with uterine preservation. Of them, 2 patients presented complications including irreducible cryptorchidism, and 3 patients developed Müllerian remnant cysts. Three patients underwent subtotal hysterectomy. Of them, one patient had complication of injury to the vas deferens, and one had hemorrhage after operation. This is the first report of PMDS involving a large Chinese population. The present study not only expands the variation spectrum but also provides clinical experience about the management of the uterus.

Analysis of clinical phenotype and genotype of Chinese children with disorders of sex development / 中华儿科杂志

Objective: To explore the heterogeneity and correlation of clinical phenotypes and genotypes in children with disorders of sex development (DSD). Methods: A retrospective study of 1 235 patients with clinically proposed DSD in 36 pediatric medical institutions across the country from January 2017 to May 2021. After capturing 277 DSD-related candidate genes, second-generation sequencing was performed to analyzed the heterogeneity and correlation combined with clinical phenotypes. Results: Among 1 235 children with clinically proposed DSD, 980 were males and 255 were females of social gender at the time of initial diagnosis with the age ranged from 1 day of age to 17.92 years. A total of 443 children with pathogenic variants were detected through molecular genetic studies, with a positive detection rate of 35.9%. The most common clinical phenotypes were micropenis (455 cases), hypospadias (321 cases), and cryptorchidism (172 cases) and common mutations detected were in SRD5A2 gene (80 cases), AR gene (53 cases) and CYP21A2 gene (44 cases). Among them, the SRD5A2 mutation is the most common in children with simple micropenis and simple hypospadias, while the AMH mutation is the most common in children with simple cryptorchidism. Conclusions: The SRD5A2 mutation is the most common genetic variant in Chinese children with DSD, and micropenis, cryptorchidism, and hypospadias are the most common clinical phenotypes. Molecular diagnosis can provide clues about the biological basis of DSD, and can also guide clinicians to perform specific clinical examinations. Target sequence capture probes and next-generation sequencing technology can provide effective and economical genetic diagnosis for children with DSD.

An elevated pretreatment serum globulin level predicts a poor prognosis of nasopharyngeal carcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the value of serum globulin levels before treatment in predicting the prognosis of patients with nasopharyngeal carcinoma (NPC).</p><p><b>METHODS</b>A total of 127 patients with non-disseminated NPC were recruited between January, 2009 and December, 2013 at Nanfang Hospital. The pretreatment serum globulin levels were analyzed with the receiver-operating characteristic (ROC) curve analysis to select the cut-off point for low and high pretreatment serum globulin levels. Kaplan-Meier and multivariable analyses were used to evaluate the predictive value of serum globulin levels.</p><p><b>RESULTS</b>The ROC curve analysis determined 30.05 g/L as the optimal cut-off value for pretreatment serum globulin level, which was significantly associated with gender (P=0.024) and N stage (P=0.016). Kaplan-Meier analysis showed that a high pretreatment serum globulin level (>30.05 g/L) significantly predicted poor progression-free survival (P=0.019), overall survival (P=0.034) and distant metastasis-free survival (P=0.049); multivariate analysis identified pretreatment serum globulin level as an independent prognostic factor for progression-free survival (HR=2.344, P=0.031).</p><p><b>CONCLUSION</b>Pretreatment serum globulin level may serve as a valuable marker to predict the prognosis of patients with NPC.</p>

Advances in the diagnosis and hormone replacement treatment of 46, XY disorders of sex development / 中华男科学杂志

Disorders of sex development (DSD) is defined as a congenital condition or atypical development of the chromosomal, gonadal, or anatomic sex. The diagnosis, gender assignment, and treatment of DSD require the guidance from experienced multidisciplinary teams. So far there has been no consensus about it in China. Due to dysgenetic gonads, defects in sex steroid biosynthesis or action, or gonadectomy during the prepubertal years, those with DSD suffer from hypogonadism. The hormone replacement therapy of DSD aims at general physiological health and long-term prognosis as well as the avoidance of unnecessary genital and gonadal surgery. This review focuses on the advances in the studies of the diagnosis and hormone replacement therapy of 46,XY DSD.

Correlation of casein kinase 2β overexpression to the metastatic ability of colorectal cancer cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of casein kinase 2β (ck2β) in colorectal cancer in relation to the metastatic ability of the cancer cells.</p><p><b>METHODS</b>The expression of ck2β in 46 normal colorectal mucosa, 20 colorectal adenomas and 66 colorectal cancers were detected immunohistochemically. In colorectal cancer cells, Ck2β protein expression was knockdown by RNA interference using ck2β-specific small interfering RNA (siRNA) and the interference efficiency was assessed by Western blotting. The effect of ck2β gene knockdown on the proliferation of the colorectal cancer cells was tested with colony formation assay, and the changes in the invasive ability of the cells were observed using Transwell chamber assay.</p><p><b>RESULTS</b>Negative or weak ck2β expression was detected in normal colorectal mucosa, with nuclear positivity in 8.7% and cytoplasmic positivity in 13.0% of the cases. Colorectal adenomas showed moderate ck2β expression, with 60% cases showing positivity in the cell nuclei and 40% in the cytoplasm. In colorectal cancers, significantly stronger expression of ck2β was found than that in colorectal adenomas and normal colorectal mucosa (P<0.05), and 75.8% cases showed positivity in the cell nuclei and 62.1% showed cytoplasmic positivity; the expression of ck2β protein in colorectal cancers with lymph node metastasis was even higher (P<0.05). Ck2β knockdown obviously inhibited the proliferation and invasiveness of colorectal cancer cells in vitro.</p><p><b>CONCLUSION</b>The high expression of ck2β in colorectal cancer is closely correlated to the carcinogenesis and metastasis of the tumor.</p>

MicroRNA profiling in patients with hepatocellular carcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the differential expression of microRNAs between human hepatocellular carcinoma (HCC) and liver cirrhosis (LC).</p><p><b>METHODS</b>The total RNA was extracted from 25 pairs of HCC and LC tissues. microRNA microarray was used to analyze the microRNA expression profiles, and validation of the microarray results was carried out by real-time quantitative RT-PCR (qRT-PCR).</p><p><b>RESULTS</b>Three microRNAs exhibited higher expression in the HCC samples than in the LC samples. In comparison with the LC samples, the HCC samples showed down-regulated expressions of 9 microRNAs. qRT-PCR verified that has-miR-122a, has-miR-199a, and has-miR-199b were downregulated in HCC, which was in agreement with the microarray results.</p><p><b>CONCLUSION</b>HCC and LC samples have significantly different microRNA expression profiles. These differentially expressed microRNAs may be involved in the pathogensis of HCC.</p>

Analysis of clinical characteristics of chronic rhinosinusitis in patients with postirradiated nasopharyngeal carcinoma / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the characters of chronic rhinosinusitis in patients with irradiated nasopharyngeal carcinoma.</p><p><b>METHODS</b>There were 65 cases of chronic rhinosinusitis after irradiated nasopharyngeal carcinoma (NPC, experimental group) and 65 cases of common chronic rhinosinusitis (CRS, control group) in the study. The visual analogue scale (VAS) was used to evaluate the intensity of subjective symptoms. Endoscopic finding was recorded and CT results were evaluated by Lund-Mackay scoring system.</p><p><b>RESULTS</b>As to the VAS, nasal secretion was significantly more severe in experimental group (7.86+/-1.62), compared with control group (5.12+/-1.32, t=10.541, P<0.01). As to endoscopic finding, middle nasal meatus were clean in 35 (53.8%) cases in experimental group, and 23 cases (35.4%) in control group (chi2=4.483, P<0.05). CT score was (7.03+/-4.63) in experiment group, and (11.42+/-3.32) in control group (t=-6.207, P<0.05). The main reason lays in lower CT score and lower involved rate of ostiomeatal complex, frontal sinus, maxillary sinus, anterior ethmoid sinus.</p><p><b>CONCLUSIONS</b>The characters of chronic rhinosinusitis in patients with irradiated nasopharyngeal carcinoma is quite different from the common CRS and different therapeutic measures should be taken.</p>

Effect of protein kinase CK2 gene silencing on radiosensitization in human nasopharyngeal carcinoma cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of protein kinase CK2 gene silencing on the radiosensitization in human nasopharyngeal carcinoma (NPC) cells and its possible mechanism.</p><p><b>METHODS</b>RNA interference (RNAi) technique was used to down-regulate the protein kinase CK2alpha expression in 5-8F cells, and clonogenic assay was employed to observe the changes in the radiosensitivity of the cells. DNA double-strand break was assessed by immunofluorescence staining of gamma-H2AX foci, and the cell apoptosis was examined using Annexin V-FITC/PI double-staining flow cytometry.</p><p><b>RESULTS</b>CK2alpha protein was successfully silenced by siRNA. CK2alpha knockdown significantly decreased the clonogenic activity and increased the radiosensitivity of the NPC cells. After a 15-min exposure of the cells to 1 Gy radiation, significant difference occurred in the gamma-H2AX foci between CK2alpha knockdown cells and the control cells (P<0.01). CK2alpha silencing significantly increased the cell apoptosis after the exposure (P<0.01).</p><p><b>CONCLUSIONS</b>Protein kinase CK2 plays an important role in the radiosensitivity of the NPC cells, and suppression of its expression might be a potential therapeutic approach of cancer.</p>

Fascin expression in human epithelial tumors and its clinical significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of FSCN1 in human epithelial tumors and their clinical significance.</p><p><b>METHODS</b>FSCN1 expression was examined immunohistochemically in specimens of human epithelial tumors, including 26 cases of lung cancer, 33 cervical cancer, 22 ovarian cancer, 38 esophageal cancer, 24 pancreatic cancer, 23 gastric cancer, 29 laryngocarcinoma, 17 primary hepatocellular carcinoma, 34 colorectal cancer, 33 breast cancer, 24 nasopharyngeal carcinoma and their corresponding normal tissues.</p><p><b>RESULTS</b>The positivity rates of FSCN1 expression in epithelial tissues and epithelial tumors were 6.3% (5/80) and 58.7% (178/303), respectively. FSCN1 showed higher expressions in cervical cancer, ovarian cancer, esophageal cancer, pancreatic cancer, gastric cancer, laryngocarcinoma, colorectal cancer, breast cancer and nasopharyngeal carcinoma, but lower or no expression in the corresponding normal tissues (P<0.05). In gastric cancer and nasopharyngeal carcinoma, the edges of the tumors were more strongly stained for FSCN1 than the interior of the tumor.</p><p><b>CONCLUSION</b>FSCN1 expression is significantly upregulated in human epithelial tumors in close correlation with tumor occurrence and progression.</p>