Phosphorylation status of CPK28 affects its ubiquitination and protein stability.

Plant innate immunity is tightly regulated. The Arabidopsis thaliana CALCIUM-DEPENDENT PROTEIN KINASE28 (CPK28) functions as a negative immune regulator. We recently demonstrate that CPK28 undergoes ubiquitination that is mediated by two ubiquitin ligases, ARABIDOPSIS TÓXICOS EN LEVADURA31 (ATL31) and ATL6, which results in its proteasomal degradation. CPK28 undergoes both intermolecular autophosphorylation and BIK1-mediated phosphorylation. However, whether the phosphorylation status of CPK28 dictates its ubiquitination and degradation is unknown yet. We used immune response analysis, transient degradation system, ubiquitination assays, co-immunoprecipitation, and other biochemical and genetic approaches to investigate the effect of the phosphorylation status of CPK28 on its degradation mediated by ATL31/6. We found the mutation of Ser318 (a site of both intermolecular autophosphorylation and BIK1-mediated phosphorylation) or a BIK1 phosphorylation site on CPK28 leads to its compromised association with ATL31 and reduced ubiquitination by ATL31. Moreover, we confirm the previous findings that two CPK28s can interact with each other, which likely promotes the intermolecular autophosphorylation. We also show that the phosphorylation status of CPK28 in turn affects its intermolecular association. We demonstrate that the phosphorylation status of CPK28 affects its degradation mediated by ATL31. Our findings reveal a link between phosphorylation of CPK28 and its ubiquitination and degradation.

Long-term survival and prognostic factors for locally advanced renal cell carcinoma with renal vein tumor thrombus.

BACKGROUND: Previous related studies have mainly focused on renal cell carcinoma (RCC) with venous tumor thrombus, specifically inferior vena cava tumor thrombus with renal vein tumor thrombus (RVTT). However, only a few studies have focused on postoperative long-term survival of RCC patients exclusively with RVTT. Our aim was to investigate the independent prognostic factors for locally advanced RCC with RVTT in China. METHODS: Patients with locally advanced RCC with RVTT were enrolled for the study from January 2000 to December 2015. All patients underwent radical nephrectomy. Survival analysis was estimated using Kaplan-Meier. Univariable and multivariable survival analyses were performed using COX. Patients were divided into high-risk, middle-risk, and low-risk groups based on independent prognostic factors and then analyzed for survival. RESULTS: One hundred twenty-eight consecutive patients (103 men & 25 women) were enrolled with a median age of 61 years. Thrombi were all graded 0 using the Mayo system, of which 23 were friable. None of the thrombi detached during surgery. 121 patients were successfully followed up, with a median follow-up period of 47 months. Median overall survival was 127 months (95%CI: 101-153). The 5-year and 10-year cancer-specific survival (CSS) rate was 67.9 and 57.0%. 59 patients had recurrence with median time of 40 months. Friable thrombus, paraneoplastic syndrome (PNS), modified Fuhrman grade 3/4 and perirenal fat invasion were independent prognostic factors (p < 0.05). The 5-year CSS for the Low-risk group (no factors) was 100%, Middle-risk group (1-2 factors) was 68.6%, while the High-risk group (3-4 factors) was 0%. CONCLUSIONS: After radical surgery, RCC patients with RVTT had a relatively fair prognosis except for patients with friable thrombus, PNS, higher modified Fuhrman grade and perirenal fat invasion.

Comparison of intracorporeal and extracorporeal urinary diversions after laparoscopic radical cystectomy in females with bladder cancer.

PURPOSE: To compare the peri-operative outcomes of females undergoing laparoscopic intracorporeal urinary diversions (ICUD) and extracorporeal urinary diversions (ECUD) after laparoscopic radical cystectomies (LRC). PATIENTS AND METHODS: Thirty-eight females who underwent LRCs and urinary diversions from February 2008 to October 2018 were divided into two groups: the ECUD group (19 patients) and the ICUD group (19 patients). We retrospectively analysed the patients in terms of patients' demographics, peri-operative outcomes, and oncological follow-ups. RESULTS: There were significant differences in the mean operative times between ECUDs and ICUDs (364.6 vs. 297.1 min, p = 0.007), transfusion rates (37% vs. 5%, p = 0.042), time to flatus (5 vs. 3 days, p = 0.020), time to ambulation (2 vs. 1 days, p = 0.022), and duration of postoperative hospital stays (22 vs. 13 days, p = 0.002). The mean lymph node yield was 12.9 in the ECUD group and 18.6 in the ICUD group (p = 0.140). Seven out of 19 patients (37%) in the ECUD group and 6 out of 19 patients (32%) in the ICUD group had positive lymph nodes (p > 0.9). Two out of 19 ECUD patients (11%) and 4 of 19 ICUD patients (21%) had positive surgical margins (p = 0.660). Although there were no differences in major complications at 30 days and in all complications at 90 days, the Clavien grade II complications were significantly different at 30 days (ECUD 8, ICUD 2; p = 0.026). The mean follow-up times were 48.7 months (ECUD group) and 26.4 months (ICUD group). There were no statistically significant differences in estimated glomerular filtration rates postoperatively (p = 0.516). Seven patients had disease metastases (ECUD 2 out of 19, ICUD 5 out of 19; p = 0.405) and 5 died (ECUD 3 out of 19, ICUD 2 out of 19; p > 0.9). CONCLUSIONS: ICUDs benefit females by having smaller incisions, faster recoveries, and decreased complication rates.

Synthesis and biological validation of N7-(4-chlorophenoxyethyl) substituted dinucleotide cap analogs for mRNA translation.

Design, synthesis and biological validation of dinucleotide cap analogs, N(7)-(4-chlorophenoxyethyl)-G(5')ppp(5')G (5a) and N(7)-(4-chlorophenoxyethyl)-m(3'-O)G(5')ppp(5')G (5b) are reported. The effect of N(7)-(4-chlorophenoxyethyl) substitution on cap analogs has been evaluated with respect to its in vitro transcription by using T7 RNA polymerase capping efficiency, and translational activity. The gel shift assay indicates that the new cap analogs (5a, 5b) showed 77% and 76% capping efficiency respectively, whereas the standard cap analog, m(7)G(5')ppp(5')G has a capping efficiency of 63%. The capping efficiency experiment clearly demonstrates that the N(7)-modified analogs are good substrate for T7 RNA polymerase. It is noteworthy that the mRNA poly(A) capped with N(7)-(4-chlorophenoxyethyl)-m(3'-O)G(5')ppp(5')G (5b) was translated ∼1.64-fold more efficiently, while compound (5a) was translated ∼0.72-fold less efficiently than mRNA capped with standard cap analog. The observed low translation activity for (5a) could be due to stability in the form of dinucleotide cap analogs. Based on the substrate compatability of the N(7) modification in dinucleotide form, these new analogs may be used for structure function studies as well as protein production.

A MAPK signaling cascade regulates the fusaric acid-induced cell death in Arabidopsis.

Mycotoxin contamination of foods and feeds is a global problem. Fusaric acid (FA) is a mycotoxin produced by Fusarium species that are phytopathogens of many economically important plant species. FA can cause programmed cell death (PCD) in several plant species. However, the signaling mechanisms of FA-induced cell death in plants are largely unknown. Here we showed that FA induced cell death in the model plant Arabidopsis thaliana, and MPK3/6 phosphorylation was triggered by FA in Arabidopsis. Both the acid nature and the radical of FA are required for its activity in inducing MPK3/6 activation and cell death. Expression of the constitutively active MKK5DD resulted in the activation of MPK3/6 and promoted the FA-induced cell death. Our work demonstrates that the MKK5-MPK3/6 cascade positively regulates FA-induced cell death in Arabidopsis and also provides insight into the mechanisms of how cell death is induced by FA in plants.

The enzymatic hydrolysate of fucoidan from Sargassum hemiphyllum triggers immunity in plants.

Fucoidans are polysaccharides that consist predominantly of sulfated L-fucoses, from which, fucoidan oligosaccharides (FOSs) are prepared through different methods. Fucoidan has versatile physiological activities, like antiviral functions against SARS CoV-2 and bioactivitiy in enhancing immune responses. Although fucoidan or FOS has been widely used in mammals as functional foods and new drugs, its application in plants is still very limited. Moreover, whether fucoidan or its derived hydrolytic products can trigger immune responses in plants remained unknown. In this work, we demonstrate that the fucoidan enzymatic hydrolysate (FEH) prepared from Sargassum hemiphyllum triggers various immune responses, such as ROS production, MAPK activation, gene expression reprogramming, callose deposition, stomatal closure, and plant resistance to the bacterial strain Pseudomonas syringae pv. tomato (Pst) DC3000. Notably, FEH did not induce Arabidopsis root growth inhibition at the concentration used for triggering other immune responses. Our work suggests that EHF can potentially be used as a non-microbial elicitor in agricultural practices to protect plants from pathogen infection.

Copy number variation of urine exfoliated cells by low-coverage whole genome sequencing for diagnosis of prostate adenocarcinoma: a prospective cohort study.

BACKGROUND: Non-invasive, especially the urine-based diagnosis of prostate cancer (PCa) remains challenging. Although prostate cancer antigen (PSA) is widely used in prostate cancer screening, the false positives may result in unnecessary invasive procedures. PSA elevated patients are triaged to further evaluation of free/total PSA ratio (f/t PSA), to find out potential clinically significant PCa before undergoing invasive procedures. Genomic instability, especially chromosomal copy number variations (CNVs) were proved much more tumor specific. Here we performed a prospective study to evaluate the diagnostic value of CNV via urine-exfoliated cell DNA analysis in PCa. METHODS: We enrolled 28 PSA elevated patients (≥ 4 ng/ml), including 16 PCa, 9 benign prostate hypertrophy (BPH) and 3 prostatic intraepithelial neoplasia (PIN). Fresh initial portion urine was collected after hospital admission. Urine exfoliated cell DNA was analyzed by low coverage Whole Genome Sequencing, followed by CNV genotyping by the prostate cancer chromosomal aneuploidy detector (ProCAD). CNVs were quantified in absolute z-score (|Z|). Serum free/total PSA ratio (f/t PSA) was reported altogether. RESULTS: In patients with PCa, the most frequent CNV events were chr3q gain (n = 2), chr8q gain (n = 2), chr2q loss (n = 4), and chr18q loss (n = 3). CNVs were found in 81.2% (95% Confidence Interval (CI) 53.7-95.0%) PCa. No CNV was identified in BPH patients. A diagnosis model was established by incorporating all CNVs. At the optimal cutoff of |Z|≥ 2.50, the model reached an AUC of 0.91 (95% CI 0.83-0.99), a sensitivity of 81.2% and a specificity of 100%. The CNV approach significantly outperformed f/t PSA (AUC = 0.62, P = 0.012). Further analyses showed that the CNV positive rate was significantly correlated with tumor grade. CNVs were found in 90.9% (95% CI 57.1-99.5%) high grade tumors and 60.0% (95% CI 17.0-92.7%) low grade tumors. No statistical significance was found for patient age, BMI, disease history and family history. CONCLUSIONS: Urine exfoliated cells harbor enriched CNV features in PCa patients. Urine detection of CNV might be a biomarker for PCa diagnosis, especially in terms of the clinically significant high-grade tumors.

Multimodal Treatment Combining Salvage Surgery-Assisted Chemotherapy and Checkpoints Blockade Immunotherapy Achieves Complete Remission on a Recurrent Penile Cancer Patient: A Case Report.

Penile squamous cell carcinoma (pSCC) is a rare disease in developed countries. pSCC causes a severe health problem and social burden in developing countries. We reported a 49-year-old male recurrent pSCC patient with medium PD-L1 expression and low TMB. The patient obtained complete response after multimodal therapy (MMT). The clinical manifestation is a recurrence in the right groin with nearly ruptured pSCC. He had partial resection of penile cancer plus bilateral groin lymph node dissection and pelvic lymph node dissection during the first operation. Pathology of the recurrent tumor showed fibrous tissue with cancer infiltration and necrosis. We used MMT, including resection of palliative right inguinal metastases, four cycles of paclitaxel+bleomycin+cisplatin, and continuous sintilimab to treat the patient. The patient had a complete response (CR) after four cycles of therapy and sustained CR for 18 months with continuous sintilimab, showing a good tolerance and acceptable toxicity. This is the first case presenting a complete response in a relapsed pSCC patient. These results suggest that MMT is worth exploring.

Neoadjuvant Chemotherapy-Guided Bladder-Sparing Treatment for Muscle-Invasive Bladder Cancer: Results of a Pilot Phase II Study.

PURPOSE: Reduced quality of life after cystectomy has made bladder preservation a popular research topic for muscle-invasive bladder cancer (MIBC). Previous research has indicated significant tumor downstaging after neoadjuvant chemotherapy (NAC). However, maximal transurethral resection of bladder tumor (TURBT) was performed before NAC to define the pathology, impacting the real evaluation of NAC. This research aimed to assess real NAC efficacy without interference from TURBT and apply combined modality therapies guided by NAC efficacy. MATERIALS AND METHODS: Patients with cT2-4aN0M0 MIBC were confirmed by cystoscopic biopsy and imaging. NAC efficacy was assessed by imaging, urine cytology, and cystoscopy with multidisciplinary team discussion. Definite responders (≤ T1) underwent TURBT plus concurrent chemoradiotherapy. Incomplete responders underwent radical cystectomy or partial cystectomy if feasible. The primary endpoint was the bladder preservation rate. RESULTS: Fifty-nine patients were enrolled, and the median age was 63 years. Patients with cT3-4 accounted for 75%. The median number of NAC cycles was three. Definite responders were 52.5%. The complete response (CR) was 10.2%, and 59.3% of patients received bladder-sparing treatments. With a median follow-up of 44.6 months, the 3-year overall survival (OS) was 72.8%. Three-year OS and relapse-free survival were 88.4% and 60.0% in the bladder-sparing group but only 74.3% and 37.5% in the cystectomy group. The evaluations of preserved bladder function were satisfactory. CONCLUSION: After stratifying MIBC patients by NAC efficacy, definite responders achieved a satisfactory bladder-sparing rate, prognosis, and bladder function. The CR rate reflected the real NAC efficacy for MIBC. This therapy is worth verifying through multicenter research.

[Combined therapy of advanced adrenal cortical adenocarcinoma].

OBJECTIVE: To analyze the clinical efficacy of combined therapy in the treatment of advanced adrenal cortical adenocarcinoma. METHODS: The clinical data of 12 cases with advanced adrenal cortical adenocarcinoma at our hospital from 1986 - 2006 were analyzed. And the relevant literatures were reviewed. RESULTS: Eleven patients underwent operation to remove primary lesions. Only 1 case received a biopsy. All cases were treated with chemotherapy or chemotherapy pulse post-operative radiotherapy. Pathological diagnosis was all of adrenal cortical adenocarcinoma. According to the staging criteria of Jacques and Brennan, all 12 cases were of IV stage. The follow-up duration was 6 - 40 months. According to evaluation criterion of chemotherapeutic effect by WHO in 1987, the results were: CR (complete remission) (n = 0), PR (partial remission) (n = 7), SD (stable disease) (n = 3) and PD (progressive disease) (n = 2). The effective rate was 58.3% (7/12). The median survival time was 14 months and median progression-free survival time 9 months. CONCLUSION: Combined therapy of adrenal cortical adenocarcinoma is effective to prolong the patient lifespan. Radiotherapy offers partial symptomatic relief for those with osseous metastasis. Making an early diagnosis and offering a novel therapy yield a better outcome.

[Diagnosis and treatment of sarcomatoid renal cell carcinoma].

OBJECTIVE: To study the clinical diagnosis and treatment of sarcomatoid renal cell carcinoma, and to analyze the connection of prognosis with therapy. METHODS: 10 cases with sarcomatoid renal cell carcinoma from 1997 - 2007 in our hospital were analyzed, and the related literatures were reviewed. RESULTS: All patients were performed with operation, and pathologically diagnosed as sarcomatoid renal cell carcinoma. Cases with metastasis were performed with chemical therapy, biological therapy or radiotherapy after operation. Clinical stage (TNM) of 10 cases is T1N0M0 1 case, T2N0M0 3 cases, T3N0M0 1 case, T4N0M0 1 case, T2N0M1 3 cases, T4N0M1 1 case. 60% (6/10) of patients were diagnosed with metastasis or diffusion. In these 6 cases, 4 cases were diagnosed with pulmonary metastasis, 1 with brain metastasis, and 1 with osseous metastasis, and 1 with inferior vena cava tumor thrombus. The average life span of all 10 cases was 17 months, and cases with advanced stage was 8 months. Otherwise, the average survival time of cases with earlier stage was 30 months, and it was 21 months from diagnosis to metastasis. CONCLUSION: Sarcomatoid renal cell carcinoma is a special type of renal cell carcinoma with features of high malignant and rapid progress. It is clinical rare and with bad prognosis. Operation is the first choice for the therapy, the effect of radiotherapy or chemical therapy is not obviously. The lung is main position for metastasis. Early diagnosis and treatment are effective to prolong the life span of patients, and the new therapy may be more important.

Whole-genome sequencing of prostate cancer reveals novel mutation-driven processes and molecular subgroups.

Prostate cancer (PCa) is the second most frequently diagnosed cancer in men. However, its genetic characteristics in the Chinese population have not been extensively profiled. Here we screened 27 Chinese patients and preformed whole-genome sequencing to dissect their genomic patterns. We found that 18.5% (5/27) tumors harbored non-protein coding mutations on FOXA1. Besides, novel focal amplifications/deletions involving ZBTB7B, SLC4A4, TBX18, CYSLTR2 and EFNA5 were frequently present in tumors. Notably, group specificity of base substitution signature B displayed a strong link to hotspot mutations on SPOP gene. Furthermore, based on six rearrangement signatures, tumors were assigned to five subgroups that revealed different biological mechanisms. Of which, tandem duplicator subgroup harbored all CDK12 mutations, small deletor subgroup owned 75% TP53 changes, and large deletor subgroup had 66.7% SPOP mutations. Taken together, we provide a comprehensive view of genomic patterns which affect the critical cell regulators of PCa in the Chinese population. Our findings may provide valuable insights for designing specific treatments for Chinese patients with PCa.

Combination of Hematology Indicators and Oncological Characteristics as a New Promising Prognostic Factor in Localized Clear Cell Renal Cell Carcinoma.

PURPOSE: This study aimed to construct a predictive model for recurrence and metastasis in patients with localized clear cell renal cell carcinoma (ccRCC) based on multiple preoperative blood indexes and oncological characteristics. PATIENTS AND METHODS: Overall, 442 patients with localized ccRCC between 2013 and 2015 were included. Using least absolute shrinkage and selection operator (LASSO) Cox regression analysis, the top three risk factors from the peripheral blood indicators were screened to construct a risk score, and a prognostic model was established. Harrell's concordance index (C-index) was applied to evaluate the predictive accuracy of the model for predicting disease-free survival (DFS) in ccRCC. RESULTS: Out of 38 blood indexes, the top three predictors were fibrinogen (FIB), C-reactive protein (CRP) and neutrophil-lymphocyte ratio (NLR). The FIB-CRP-NLR (FCN) score (hazard ratio [HR]: 1.86, 95% confidence interval [CI]: 1.21-2.9, P = 0.005) was an independent prognostic factor in multivariate analysis. Furthermore, the FIB-CRP-NLR-T-Grade (FCNTG) risk model combining FCN score, T stage and Furhman grade achieved a higher prognostic accuracy (mean C-index, 0.728) than both the FCN score alone (mean C-index, 0.675) and the stage, size, grade, and necrosis (SSIGN) score (mean C-index, 0.686) in the validation cohort. CONCLUSION: The FCN score combining peripheral blood indicators of inflammation and coagulation is an independent prognostic marker of ccRCC. The FCNTG model, which systemically incorporates preoperative blood indexes to oncological characteristics, shows its advantages of convenience and high prediction efficiency.

Efficacy and Safety of Androgen-Deprivation Therapy Combined with Docetaxel Plus Prednisone in High-Burden Metastatic Hormone-Sensitive Prostate Cancer.

PURPOSE: The aim of this study was to evaluate the efficacy and safety of hormonal and synchronous docetaxel plus prednisone (DocP) in metastatic hormone-sensitive prostate cancer (mHSPC). METHODS: One hundred fifty-one cases with high-burden mHSPC diagnosed at 1 single center from January 2014 to August 2018 were analyzed retrospectively. Among them, 85 cases received androgen-deprivation therapy (ADT) within 3 months, along with 6 cycles of docetaxel + prednisone (treatment group), whereas 66 received ADT alone (control group). The primary end point was the median overall survival (OS), while the secondary outcomes included prostate-specific antigen (PSA) progression-free survival (PFS), radiographic PFS, and the proportion of PSA falling to 0.2 ng/mL. RESULTS: A total of 151 patients were included and followed up for a median of 34 months in this study. The median OS time in the treatment group was unavailable, but it was remarkably longer than that of the control group (P<0.001). In addition, the PFS of PSA in the treatment group and control group was 17.9 months and 9.2 months, respectively (P<0.001). Meanwhile, the radiographic PFS was 43 months in the treatment group and 19.8 months in the control group, respectively (P<0.001). The proportions of PSA falling to 0.2 ng/mL were 53.7% and 23.3%, respectively (P<0.001). However, there was no significant difference in the incidence of ≥3 toxic side effects between these 2 groups (P=0. 21). CONCLUSION: ADT combined with 6 cycles of docetaxel + prednisone chemotherapy benefits patients diagnosed with high-burden mHSPC in terms of the OS, PFS of PSA and radiographic, and the ratio of PSA falling to 0.2 ng/mL.

Comparison of Laparoscopy and Open Radical Nephrectomy of Renal Cell Cancer.

OBJECTIVE: The aim of this study was to summarize the current evidence to evaluate the effects of laparoscopic radical nephrectomy (LRN) and open radical nephrectomy (ORN) in the treatment of renal cell carcinoma. METHODS: A comprehensive literature search was performed using PubMed, Embase and Google Scholar to identify all relevant studies. 8 published studies were included in this meta-analysis. We pooled the odds ratios (OR), standardised mean difference (SMD) and conducted heterogeneity, and quality assessment. RESULTS: The outcome of treatment effects included surgical blood loss, surgical time, postoperative complications, and post-operative length of hospital stay. Comparing open radical nephrectomy for kidney cancer patients, the pooled SMD of surgical time was 0.47, (95% confidential index CI = [0.09, 0.85]), the pooled SMD of operative blood loss was -68.98, (95% CI = [-99.63, -38.34]), the pooled SMD of post-operative length of hospital stay was -4.32, (95% CI = [-4.62, -4.03]), and the pooled OR of postoperative complications was 0.52, (95% CI = [0.30, 0.91]). CONCLUSION: LRN was found to significantly decrease patients' blood loss. In addition, LRN offers less post-operative length of hospital stay.

Feasibility of prostatectomy without prostate biopsy in the era of new imaging technology and minimally invasive techniques.

BACKGROUND: Routinely, after receiving prostate specific antigen (PSA) testing and digital rectum examination, patients with suspected prostate cancer are required to undergo prostate biopsy. However, the ability of ultrasound-guided prostate biopsy to detect prostate cancer is limited. Nowadays, a variety of diagnostic methods and more sensitive diagnostic methods, such as multi-parameter prostate magnetic resonance imaging (mpMRI) and prostate-specific membrane antigen positron emission tomography/computed tomography (PSMA PET/CT) can be applied clinically. Furthermore, laparoscopic/robot-assisted prostatectomy is also a safe and effective procedure for the treatment of benign prostatic hyperplasia. So maybe it is time to reconsider the necessary to perform prostate biopsy before radical prostatectomy. AIM: To explore the feasibility of radical prostatectomy without prostate biopsy in the era of new imaging technology and minimally invasive techniques. METHODS: From June 2014 to November 2018, 11 cases of laparoscopic radical prostatectomy without prostate biopsy were performed at the three tertiary medical centers involved in this study. All patients received prostate magnetic resonance imaging and prostate cancer was suspected, including six patients with positive 68Ga-PSMA PET/CT results. Laparoscopic radical prostatectomy and pelvic lymph node dissection were performed for all patients. RESULTS: All surgeries were accomplished successfully. The mean age was 69 ± 7.7 year, the mean body mass index was 24.7 ± 1.6 kg/m2, the range of serum PSA was 4.3 to >1000 ng/mL, and the mean prostate volume was 40.9 ± 18.3 mL. The mean operative time was 96 ± 23.3 min, the mean estimated blood loss was 90 ± 90.9 mL, and the median duration of catheter placement was 14 d. The final pathology confirmed that all specimens were prostate cancer except one case of benign prostatic hyperplasia. No major complications occurred in 90 d postoperatively. CONCLUSION: The current practice of mandating a prostatic biopsy before prostatectomy should be reconsidered in the era of new imaging technology and minimally invasive techniques. Radical prostatectomy could be carried out without the evidence of malignancy. Large-sample randomized controlled trials are definitely required to confirm the feasibility of this new concept.

[Diagnosis and treatment of malignant bladder non-epithelial tumors].

OBJECTIVE: To study the clinical diagnosis, treatment, and prognosis of malignant bladder non-epithelial tumors. METHODS: The clinical data of 17 cases with malignant bladder non-epithelial tumor, 15 males and 3 females, aged 28 (3-73), were analyzed. RESULTS: Ten of the 17 cases were diagnosed as with rhabdomyosarcoma, 2 with malignant lymphoma, and 2 with malignant pheochromocytoma, 2 with leiomyosarcoma, and 1 with carcinosarcoma. All patients underwent operation, or were treated with radiotherapy or chemotherapy. Among the 10 cases with rhabdomyosarcoma, 7 were aged under 5, 5 of them survived for more than 8 years, and 2 were lost to follow-up; other 3 adult patients died in one year with metastasis all over the body. Two patients with malignant lymphoma underwent partial cystectomy and adjuvant radiotherapy or chemotherapy, and survived for more than 3 years. Of the 2 cases with malignant pheochromocytoma, one underwent lymphadenectomy and adjuvant chemotherapy and survived for more than 5 years; and another case died 2 years after operation and chemotherapy. Two patients with leiomyosarcoma underwent simple resection, one survived for more than 5 years; and another survived 1 year after operation. One case with carcinosarcoma receiving operation followed by chemotherapy died in one year. CONCLUSION: Malignant bladder non-epithelial tumors are rare clinically and most of them occur in children. Most of them are rhabdomyosarcoma. The prognosis of malignant bladder non-epithelial tumors, different in pathological types, is relatively worse in adults.

An Efficient Protection-Free One-Pot Chemical Synthesis of Modified Nucleoside-5'-Triphosphates.

A simple, reliable, and an efficient "one-pot, three step" chemical method for the synthesis of modified nucleoside triphosphates such as 5-methylcytidine-5'-triphosphate (5-MeCTP), pseudouridine-5'-triphosphate (pseudoUTP) and N(1)-methylpseudouridine-5'-triphosphate (N(1)-methylpseudoUTP) starting from the corresponding nucleoside is described. The overall reaction involves the monophosphorylation of nucleoside, followed by the reaction with pyrophosphate and subsequent hydrolysis of the cyclic intermediate to furnish the corresponding NTP in moderate yields with high purity (>99.5%).

[Expression of Survivin in transitional cell carcinoma of urinary bladder and its clinical significance].

OBJECTIVE: Survivin is a member of the inhibitors of apoptosis protein (IAP) family. Recent researches had shown that survivin plays an important role in oncogenesis. This study was designed to investigate the expression of survivin in transitional cell carcinoma (TCC) of urinary bladder and its clinical significance. METHODS: Immunohistochemical assay was used to detect the expression of survivin in 75 cases of tumor tissue and 7 cases of normal bladder tissue. RESULTS: The expression rates of survivin were 77.3% (58/75) in TCC of urinary bladder, whereas no expression of survivin was detected in the 7 cases of normal bladder tissue. CONCLUSIONS: Expression of survivin protein was observed in the tumor tissue derived from the patients with bladder TCC, indicating that this protein may play an important rule in carcinogenesis of human urinary bladder. The expression of survivin was statistically significant associated with tumor grade. Our results suggested that the expression of survivin may be considered as a prognostic factor for bladder TCC.

[Loss of heterozygosity of tumor suppressor genes at chromosome 3p in transitional cell carcinoma of urinary bladder].

OBJECTIVE: To investigate the potential effects of three tumor suppressor genes located at chromosome 3p, FHIT, hMLH1 and VHL, in carcinogenesis of transitional cell carcinoma (TCC) of urinary bladder in Chinese. METHODS: PCR was used to examine the heterozygosity and loss of heterozygosity (LOH) of the 6 microsatellite polymorphic markers inside of or flanking to the three genes tumor suppressor genes FHIT, hMLHi, and VHL in TCC tissues of 40 cases. RESULTS: The rates of heterozygosity and LOH were 55.0% (38/40) and 59.1% (22/38) for hMLH1 gene. The rates of heterozygosity and LOH were 90.0% (36/40) and 47.2% (17/36). The rate of heterozygosity of the two microsatellite polymorphic markers in the intron of FHIT gene, D2S1234 and D3S1300, were 70% (28/40) and 67.5% (27/40), The LOH rates of D2S1234 and D3S1300 were 46.4% (13/28) and 37% (10/27) respectively. The rate that at least one the these 2 microsatellite polymorphic markers was heterozygote was 95.0% (38/40), the rate that LOH occurred for at least one of them was 57.8% (22/38). The heterozygosity rate of D3S1561 and D3S1612, two microsatellite polymorphic markers linked to hMLH1 gene, were 42.5% (17/40) and 30% (12/40) respectively, the LOH rates for these two markers were 41.2% (7/17) and 75.0% (9/12) respectively, the rate that at least one of these two markers was heterozygote was 59.1% (13/22). The heterozygosity rate of D3S10368 and D3S1284, two microsatellite polymorphic markers linked to VHL gene, were 70% (28/40) and 47.5% (19/40) respectively, the LOH rates for these two markers were 42.8% (12/28) and 42.1% (8/19) respectively, the rate that at least one of these two markers was heterozygote was 90.0% (36/40). High frequencies of LOH were found in tumor suppressor genes FHIT, hMLH1 and VH. Only the LOH of D3S1284 was positively correlated with the pathological staging of TCC of urinary bladder (P < 0.05). CONCLUSION: LOH in tumor suppressor genes FHIT, hMLH1 and VHL may play an important role in carcinogenesis of human urinary bladder, and may provide new approaches for early detection of TCC. Loss of VHK gene may be a late event of carcinogenesis of TCC.