Single-cell and spatial transcriptomics reveal 5-methylcytosine RNA methylation regulators immunologically reprograms tumor microenvironment characterizations, immunotherapy response and precision treatment of clear cell renal cell carcinoma.

Clear cell Renal Cell Carcinoma (ccRCC) is a highly heterogeneous disease, making it challenging to predict prognosis and therapy efficacy. In this study, we aimed to explore the role of 5-methylcytosine (m5C) RNA modification in ccRCC and its potential as a predictor for therapy response and overall survival (OS). We established a novel 5-methylcytosine RNA modification-related gene index (M5CRMRGI) and studied its effect on the tumor microenvironment (TME) using single-cell sequencing data for in-depth analysis, and verified it using spatial sequencing data. Our results showed that M5CRMRGI is an independent predictor of OS in multiple datasets and exhibited outstanding performance in predicting the OS of ccRCC. Distinct mutation profiles, hallmark pathways, and infiltration of immune cells in TME were observed between high- and low-M5CRMRGI groups. Single-cell/spatial transcriptomics revealed that M5CRMRGI could reprogram the distribution of tumor-infiltrating immune cells. Moreover, significant differences in tumor immunogenicity and tumor immune dysfunction and exclusion (TIDE) were observed between the two risk groups, suggesting a better response to immune checkpoint blockade therapy of the high-risk group. We also predicted six potential drugs binding to the core target of the M5CRMRGI signature via molecular docking. Real-world treatment cohort data proved once again that high-risk patients were appropriate for immune checkpoint blockade therapy, while low-risk patients were appropriate for Everolimus. Our study shows that the m5C modification landscape plays a role in TME distribution. The proposed M5CRMRGI-guided strategy for predicting survival and immunotherapy efficacy, we reported here, might also be applied to more cancers other than ccRCC.

Identification of mRNA vaccines and conserved ferroptosis related immune landscape for individual precision treatment in bladder cancer.

Background: The aim of this study was to identify the ferroptosis induced tumor microenvironment (FeME) landscape in bladder cancer (BCa) for mRNA vaccine development and selecting suitable patients for precision treatment. Methods: Gene expression profiles and clinical information of 1216 BCa patients were extracted from TCGA-BLCA, three GEO databases and IMvigor210 cohort. We comprehensively established the FeME landscape of 1216 BCa samples based on 290 ferroptosis related genes (FRGs), and systematically correlated these regulation patterns with TME cell-infiltrating characteristics. Besides, we identified the patients' ferroptosis risk index (FRI) to predict the prognosis of BCa for precise treatment. Results: Six over-expressed and mutated tumor antigens associated with poor prognosis and infiltration of antigen presenting cells were identified in BCa. Furthermore, we demonstrated the evaluation of FeME within individual tumors could predict stages of tumor inflammation, subtypes, genetic variation, and patient prognosis. Then, 5-lncRNA signature was mined to produce the FRI. Low FRI was also linked to increased mutation load, better prognosis and enhanced response to anti-PD-L1 immunotherapy. Besides, an immunotherapy cohort confirmed patients with lower FRI demonstrated significant therapeutic advantages and clinical benefits. Conclusions: TFRC, SCD, G6PD, FADS2, SQLE, and SLC3A2 are potent antigens for developing anti-BCa mRNA vaccine. Establishment of FRI will contribute to enhancing our cognition of TME infiltration characterization and guiding more effective immunotherapy strategies and selecting appropriate patients for tumor vaccine therapy. Supplementary Information: The online version contains supplementary material available at 10.1186/s40537-022-00641-z.

The performance of transrectal ultrasound in the diagnosis of seminal vesicle defects: a comparison with magnetic resonance imaging.

Obstructive azoospermia (OA) is one of the most common causes of male infertility. Transrectal ultrasound (TRUS) has been used to diagnose OA for many years. From 2009 to 2013, we evaluated a prospective cohort of 1249 patients with suspected OA using TRUS. It was found that dilation of the ejaculatory duct (ED) (29.9%, 374/1249) was the most common cause of OA, followed by seminal vesicle (SV) abnormalities (28.5%, 356/1249). A total of 237 patients were diagnosed with congenital defects (agenesis and/or hypoplasia) of the SV, constituting more than half of the cases of SV disease in OA (19.0%, 237/1249). In contrast to ED, congenital defects of the SV could not be corrected with surgical treatment. Therefore, it is meaningful to compare TRUS and magnetic resonance imaging (MRI) for accurate diagnosis of SV defects. Among our patients, 30 with agenesis or/and hypoplasia of the SV on TRUS were further evaluated using pelvic MRI within 2 years, with the objective of verifying the TRUS results. The concordance rate for diagnosing congenital defects of the SV was 73.3% (22/30). We concluded that TRUS is a reliable and convenient method for diagnosing agenesis or hypoplasia of the SV in OA patients with a high concordance with MRI while MRI is useful in patients with inconclusive TRUS findings.

[Transrectal ultrasonography in the etiological diagnosis of male obstructive azoospermia: analysis of 695 cases].

OBJECTIVE: To assess the role of transrectal ultrasonography (TRUS) in the etiological diagnosis of male obstructive azoospermia. METHODS: We retrospectively analyzed the clinical data and TRUS findings of 695 patients with obstructive azoospermia from January 2007 to May 2009. RESULTS: Concerning the etiology of obstructive azoospermia, the main TRUS findings included ejaculatory duct abnormality (29.2%), seminal vesicle abnormality (25.4%) and prostate midline cyst (18.5%). TRUS revealed 203 cases of ejaculatory duct dilation, 177 cases of seminal vesicle abnormality (including 108 with absence or agenesis and 51 with dilation of the seminal vesicle), and 128 cases of prostate midline cyst (including 75 with ejaculatory duct cyst and 39 with Müllerian cyst). Calcification of the verumontanum or ejaculatory duct was suspected to be the causes of obstructive azoospermia in 34 cases. However, no significant etiological abnormality was found in 153 cases. Obvious etiology was shown by TRUS in 78.0% of the patients. CONCLUSION: TRUS can clearly display the structural abnormality of the ejaculatory duct and seminal vesicle, and provide important information on the etiology of male obstructive azoospermia.

Study on the effects of allitride and its mechanisms on renal cell carcinoma in vitro.

OBJECTIVE: To study the killing effect of allitride on human renal cell carcinoma cell line Ketr-3 and its possible mechanisms. METHODS: The Ketr-3 cells were treated with allitride and the morphological changes were observed with inverted microscope. The cytotoxicity was estimated through theamine blue tetrazolium bromide (MTT). Apoptotic cells were detected by in situ cell apoptosis detection kit, and confirmed by flow cytometry. Changes of apoptosis rate cell cycle were assessed by flow cytometry. Caspase-3 (cysteineaspartate specific proteinase) mRNA was detected by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR), and Caspase-3 protease activity was estimated with colorimetry. RESULTS: MTT assay and morphological changes confirmed the killing effect of allitride on Ketr-3 cellline. FCM also showed that S-phase and G2/M-phase arrest was induced. RT-PCR and colorimetry confirmed that there was apparently a rise of Caspase-3 mRNA and Caspase-3 protease activity. CONCLUSION: Allitride could kill Ketr-3 effectively by inducing apoptosis. Cell cycle arrest and up-regulation of Caspase-3 may play an important role in the mechanisms of killing effect.

[Value of transrectal ultrasonography in the diagnosis of midline prostatic cysts].

OBJECTIVE: To assess the value of transrectal ultrasonography (TRUS) in the diagnosis of midline prostatic cysts. METHODS: We retrospectively analyzed the TRUS manifestations of 87 cases of midline prostatic cysts. RESULTS: Of the total number, 33 cases were diagnosed as Müllerian duct cysts, 21 cases ejaculatory duct cysts and the other 33 cases undifferentiated midline prostatic cysts; 19 cases had dilated seminal vesicles, 19 seminal vesicle agenesis, 9 seminal vesiculitis and 5 dilation of the ejaculatory duct. CONCLUSION: TRUS, convenient, sensitive, safe and non-invasive, is a desirable method for the diagnosis of midline prostatic cysts.

[Individualization of transrectal ultrasonography-guided prostate biopsy for prostate cancer detection].

BACKGROUND & OBJECTIVE: Transrectal ultrasonography (TRUS)-guided sextant biopsy technique was regarded as golden standard method for the diagnosis of prostate cancer. Recently, many reports show that the detection rate of prostate cancer by sextant biopsy is not high, and suggest to take more cores to improve the detection rate. But there is no ideal protocol now. This study was to explore an appropriate prostate biopsy protocol for the detection of prostate cancer. METHODS: Clinical data of 325 consecutive men with suspected prostate cancer were analyzed. All patients underwent 12-core biopsy protocol (first biopsy) with additional 1 or 2 cores at each suspicious area detected by TRUS. The sensitivity of different combinations of biopsy cores was analyzed. RESULTS: Of the 325 patients, 126 (38.8%) were positive for prostate cancer. The detection rate by 12-core protocol was significantly higher than maximal detection rate by 6-, 8-, and 10-core protocols (38.8% vs. 27.7%, 29.8%, and 35.4%, P<0.05). In the patients with prostate volume of <40 ml, there was no significant difference in detection rate of prostate cancer between 8-, 10-, and 12-core protocols. In the patients with prostate volume of 40-60 ml, the detection rate by 10-core protocol was significantly higher than that by 8-core protocol (36.2% vs. 26.9%, P=0.046). In the patients with prostate volume of >60 ml, the detection rate by 12-core protocol was significantly higher than that by 10-core protocol (37.9% vs. 25.8%, P=0.049). CONCLUSIONS: Individual prostate biopsy protocol should be taken for the detection of prostate cancer. We recommend 8-core protocol for the patients with small prostate (<40 ml), 10-core protocol for the patients with the prostate of 40-60 ml, and 12-core protocol for the patients with the prostate of >60 ml, and add 1 or 2 cores or take focus biopsy protocol at suspicious areas detected by TRUS.

[Sonographic features and clinical significance of transrectal ultrasonography in obstructive azoospermia].

OBJECTIVE: To investigate the ultrasonographic features of obstructive azoospermia and to evaluate transrectal ultrasonography in the diagnosis of the disease. METHODS: We performed transrectal ultrasonography for 248 patients with obstructive azoospermia, observed the sonographic features of the prostate gland, seminal vesicle and ejaculatory duct. RESULTS: The average volume of the prostate gland of the studied group was 13.2 ml. A total of 111 cases showed normal sonographic features, 39 cases bilateral seminal vesicle defect, 33 cases bilateral seminal vesicle aplasia, 23 cases unilateral seminal vesicle defect and contralateral seminal vesicle aplasia, 28 cases bilateral and 14 cases unilateral seminal vesicle dilatation. Of the 42 cases of seminal vesicle dilatation, 18 had ejaculatory duct dilatation and 17 had cysts in the midline of the prostate. CONCLUSION: Obstructive azoospermia varies in kind. Transrectal ultrasonography can reveal the details of the prostate, seminal vesicle and ejaculatory duct and help to classify obstructive azoospermia and determine the location of the lesion.

[Surgical therapy for azoospermia with ejaculatory duct obstruction].

OBJECTIVE: To investigate the effect of transurethral resection of ejaculatory ducts (TURED) for azoospermia with ejaculatory duct obstruction (EDO). METHODS: From June 2003 to December 2004, 20 azoospermia with EDO were diagnosed, diagnostic criteria included a history, physical examination, semen analyses, semen fructose measurement, endocrine assessment, testicular biopsy and transrectal ultrasonography (TRUS); All 20 cases were treated by TURED. Fifteen of them were followed up more than 3 months after the treatment. The semen samples of them were analysed at 3-month intervals in post-therapy. RESULTS: Semen analyses in all 20 cases showed the typical characteristics of EDO, low semen volume (0.4-1.6 ml), azoospermia, low pH, absent or low semen fructose. TRUS showed the main etiology factor of EDO was a midline cyst in 11, lateral cystic lesions in 2, the remaining 7 cases had dilated ejaculatory duct with or without dilated seminal vesicles. Among 15 cases followed up more than 3 months after TURED, 10/15 (67%) had an improvement in semen parameters and 3/15 (20%) had pregnancies. Semen analyses had not been done in anther 5 cases. CONCLUSION: Transurethral resection of ejaculatory ducts may be a safe and effective method for the treatment of azoospermia with EDO.

[Diagnosis and treatment of renal cell carcinoma with vena cava tumor thrombi].

BACKGROUND & OBJECTIVE: Renal cell carcinoma (RCC) might involve the renal vein, and form tumor thrombus extending into the vena cava or the right atrium. Treating RCC with vena cava involvement is difficult in clinical practice. Radical nephrectomy with complete tumor thrombus removal could result in good outcomes for RCC patients with vena cava thrombi. This paper was to report our experiences on treating RCC with vena cava thrombi. METHODS: From May 1995 to Oct. 2003, radical nephrectomy plus vena cava thrombus removal was performed in 14 RCC patients. Clinical records, including preoperative diagnosis, operation pattern, and prognosis, of these 14 patients were analyzed retrospectively. RESULTS: Vena cava thrombi were detected in 9 patients, and missed in 5 patients by ultrasonography. CT scan revealed vena cava thrombi in 12 of 14 patients. MRI has been performed in 8 patients, and clearly demonstrated extent of the thrombus. Twelve cases of tumor thrombi within infrahepatic vena cava, 1 within intrahepatic subphrenic vena cava, and 1 within supraphrenic vena cava. The patients have been followed up for 6-70 months after surgery. Thirteen patients survived with disease-free, and 1 patient (stage IIIc) died of cancer 23 months after surgery. CONCLUSION: Vena cava thrombi in patients with RCC could be detected on CT scan and ultrasonography. MRI is more accurate than CT,and ultrasonography in delineating extent of the thrombus. Radical nephrectomy plus vena cava thrombus removal could achieve long-term survival for patients with localized RCC and vena cava thrombi.