Surface ligand-regulated renal clearance of MRI/SPECT dual-modality nanoprobes for tumor imaging.

BACKGROUND: The general sluggish clearance kinetics of functional inorganic nanoparticles tend to raise potential biosafety concerns for in vivo applications. Renal clearance is a possible elimination pathway for functional inorganic nanoparticles delivered through intravenous injection, but largely depending on the surface physical chemical properties of a given particle apart from its size and shape. RESULTS: In this study, three small-molecule ligands that bear a diphosphonate (DP) group, but different terminal groups on the other side, i.e., anionic, cationic, and zwitterionic groups, were synthesized and used to modify ultrasmall Fe3O4 nanoparticles for evaluating the surface structure-dependent renal clearance behaviors. Systematic studies suggested that the variation of the surface ligands did not significantly increase the hydrodynamic diameter of ultrasmall Fe3O4 nanoparticles, nor influence their magnetic resonance imaging (MRI) contrast enhancement effects. Among the three particle samples, Fe3O4 nanoparticle coated with zwitterionic ligands, i.e., Fe3O4@DMSA, exhibited optimal renal clearance efficiency and reduced reticuloendothelial uptake. Therefore, this sample was further labeled with 99mTc through the DP moieties to achieve a renal-clearable MRI/single-photon emission computed tomography (SPECT) dual-modality imaging nanoprobe. The resulting nanoprobe showed satisfactory imaging capacities in a 4T1 xenograft tumor mouse model. Furthermore, the biocompatibility of Fe3O4@DMSA was evaluated both in vitro and in vivo through safety assessment experiments. CONCLUSIONS: We believe that the current investigations offer a simple and effective strategy for constructing renal-clearable nanoparticles for precise disease diagnosis.

Single-cell transcriptomic analysis of tumor heterogeneity and intercellular networks in human urothelial carcinoma.

BACKGROUND: Heterogeneity of tumor cells and the tumor microenvironment (TME) is significantly associated with clinical outcomes and treatment responses in patients with urothelial carcinoma (UC). Comprehensive profiling of the cellular diversity and interactions between malignant cells and TME may clarify the mechanisms underlying UC progression and guide the development of novel therapies. This study aimed to extend our understanding of intra-tumoral heterogeneity and the immunosuppressive TME in UC and provide basic support for the development of novel UC therapies. METHODS: Seven patients with UC were included who underwent curative surgery at our hospital between July 2020 and October 2020. We performed single-cell RNA sequencing (scRNA-seq) analysis in seven tumors with six matched adjacent normal tissues and integrated the results with two public scRNA-seq datasets. The functional properties and intercellular interactions between single cells were characterized, and the results were validated using multiplex immunofluorescence staining, flow cytometry, and bulk transcriptomic datasets. All statistical analyses were performed using the R package with two-sided tests. Wilcoxon-rank test, log-rank test, one-way analysis of variance test, and Pearson correlation analysis were used properly. RESULTS: Unsupervised t-distributed stochastic neighbor embedding clustering analysis identified ten main cellular subclusters in urothelial tissues. Of them, seven urothelial subtypes were noted, and malignant urothelial cells were characterized with enhanced cellular proliferation and reduced immunogenicity. CD8 + T cell subclusters exhibited enhanced cellular cytotoxicity activities along with increased exhaustion signature in UC tissues, and the recruitment of CD4 + T regulatory cells was also increased in tumor tissues. Regarding myeloid cells, coordinated reprogramming of infiltrated neutrophils, M2-type polarized macrophages, and LAMP3 + dendritic cells contribute to immunosuppressive TME in UC tissues. Tumor tissues demonstrated enhanced angiogenesis mediated by KDR + endothelial cells and RGS5 + /ACTA2 + pericytes. Through deconvolution analysis, we identified multiple cellular subtypes may influence the programmed death-ligand 1 (PD-L1) immunotherapy response in patients with UC. CONCLUSION: Our scRNA-seq analysis clarified intra-tumoral heterogeneity and delineated the pro-tumoral and immunosuppressive microenvironment in UC tissues, which may provide novel therapeutic targets.

A clinically translatable kit for MRI/NMI dual-modality nanoprobes based on anchoring group-mediated radiolabeling.

Magnetic resonance imaging (MRI)/nuclear medicine imaging (NMI) dual-modality imaging based on radiolabeled nanoparticles has been increasingly exploited for accurate diagnosis of tumor and cardiovascular diseases by virtue of high spatial resolution and high sensitivity. However, significant challenges exist in pursuing truly clinical applications, including massive preparation and rapid radiolabeling of nanoparticles. Herein, we report a clinically translatable kit for the convenient construction of MRI/NMI nanoprobes relying on the flow-synthesis and anchoring group-mediated radiolabeling (LAGMERAL) of iron oxide nanoparticles. First, homogeneous iron oxide nanoparticles with excellent performance were successfully obtained on a large scale by flow synthesis, followed by the surface anchoring of diphosphonate-polyethylene glycol (DP-PEG) to simultaneously render the underlying nanoparticles biocompatible and competent in robust labeling of radioactive metal ions. Moreover, to enable convenient and safe usage in clinics, the DP-PEG modified nanoparticle solution was freeze-dried and sterilized to make a radiolabeling kit followed by careful evaluations of its in vitro and in vivo performance and applicability. The results showed that 99mTc labeled nanoprobes are effectively obtained with a labeling yield of over 95% in 30 minutes after simply injecting Na[99mTcO4] solution into the kit. In addition, the Fe3O4 nanoparticles sealed in the kit can well stand long-term storage even for 300 days without deteriorating the colloidal stability and radiolabeling yield. Upon intravenous injection of the as-prepared radiolabeled nanoprobes, high-resolution vascular images of mice were obtained by vascular SPECT imaging and magnetic resonance angiography, demonstrating the promising clinical translational value of our radiolabeling kit.

Genome-wide comparison and in silico analysis of splicing factor SYF2/NTC31/p29 in eukaryotes: Special focus on vertebrates.

The gene SYF2-an RNA splicing factor-can interact with Cyclin D-type binding protein 1 (GICP) in many biological processes, including splicing regulation, cell cycle regulation, and DNA damage repair. In our previous study we performed genome-wide identification and functional analysis of SYF2 in plant species. The phylogenetic relationships and expression profiles of SYF2 have not been systematically studied in animals, however. To this end, the gene structure, genes, and protein conserved motifs of 102 SYF2 homologous genes from 91 different animal species were systematically analyzed, along with conserved splicing sites in 45 representative vertebrate species. A differential comparative analysis of expression patterns in humans and mice was made. Molecular bioinformatics analysis of SYF2 showed the gene was conserved and functional in different animal species. In addition, expression pattern analysis found that SYF2 was highly expressed in hematopoietic stem cells, T cells, and lymphoid progenitor cells; in ovary, lung, and spleen; and in other cells and organs. This suggests that changes in SYF2 expression may be associated with disease development in these cells, tissues, or organs. In conclusion, our study analyzes the SYF2 disease resistance genes of different animal species through bioinformatics, reveals the relationship between the SYF2 genotype and the occurrence of certain diseases, and provides a theoretical basis for follow-up study of the relationship between the SYF2 gene and animal diseases.

METTL3 promotes prostate cancer progression by regulating miR-182 maturation in m6A-dependent manner.

METTL3 was known to run through the whole cycle of RNA. It relied on m6A modification in the mRNAs of cancer-related genes to regulate tumour progression. The development of prostate cancer cells could be promoted by METTL3 via hedgehog pathway. Recent studies had shown that the effect of METTL3 on non-coding RNA was mainly dependent on the modification of m6A. However, it is still unknown whether METTL3 promotes tumour development through this mechanism in prostate cancer. The expression of METTL3 in prostate cancer tissues and cells was analysed by qRT-PCR and Western blot assays. CCK-8 assay, colony formation assay, wound-healing assay and transwell assays were conducted to detect the impact of METTL3 on cell proliferation, migration and invasion. Nude mice tumour models were built to evaluate the role of METTL3 in tumorigenesis. N6-methyladenosine (m6A) RNA immunoprecipitation assay (MeRIP) and co-immunoprecipitations assays were performed to verified that METTL3 upregulated the m6A level, interacted with microprocessor protein DGCR8, recognized the m6A modification of pre-miR-182 to regulate its maturation.METTL3 was highly expressed in prostate cancer, and knockdown of METTL3 significantly inhibited cell proliferation, migration, invasion and tumorigenesis, while overexpression of METTL3 promoted cell proliferation, migration, invasion and tumorigenesis in PCa. In addition, we found that METTL3 upregulating the level of m6A, and interacted with DGCR8 to recognize the m6A modification of pre-miR-182 to regulate its splicing and maturation and promote the high expression of miRNA. Our study suggests that METTL3 could be used in targeted therapies for PCa.

Anchoring Group-Mediated Radiolabeling of Inorganic Nanoparticles─A Universal Method for Constructing Nuclear Medicine Imaging Nanoprobes.

Nuclear medicine imaging has aroused great interest in the design and synthesis of versatile radioactive nanoprobes, while most of the methods developed for radiolabeling nanoprobes are difficult to satisfy the criteria of clinical translation, including easy operation, mild labeling conditions, high efficiency, and high radiolabeling stability. Herein, we demonstrated the universality of a simple but efficient radiolabeling method recently developed for constructing nuclear imaging nanoprobes, that is, ligand anchoring group-mediated radiolabeling (LAGMERAL). In this method, a diphosphonate-polyethylene glycol (DP-PEG) decorating on the surface of inorganic nanoparticles plays an essential role. In principle, owing to the strong binding affinity to a great variety of metal ions, it can not only endow the underlying nanoparticles containing metal ions including some main group metal ions, transition metal ions, and lanthanide metal ions with excellent colloidal stability and biocompatibility but also enable efficient radiolabeling through the diphosphonate group. Based on this assumption, inorganic nanoparticles such as Fe3O4 nanoparticles, NaGdF4:Yb,Tm nanoparticles, and Cu2-xS nanoparticles, as representatives of functional inorganic nanoparticles suitable for different imaging modalities including magnetic resonance imaging (MRI), upconversion luminescence imaging (UCL), and photoacoustic imaging (PAI), respectively, were chosen to be radiolabeled with different kinds of radionuclides such as SPECT nuclides (e.g., 99mTc), PET nuclides (e.g., 68Ga), and therapeutic SPECT nuclides (e.g., 177Lu) to demonstrate the reliability of the LAGMERAL approach. The experimental results showed that the obtained nanoprobes exhibited high radiolabeling stability, and the whole radiolabeling process had negligible impacts on the physical and chemical properties of the initial nanoparticles. Through passive targeting SPECT/MRI of glioma tumor, active targeting SPECT/UCL of colorectal cancer, and SPECT/PAI of lymphatic metastasis, the outstanding potentials of the resulting radioactive nanoprobes for sensitive tumor diagnosis were demonstrated, manifesting the feasibility and efficiency of LAGMERAL.

Rapidly liver-clearable rare-earth core-shell nanoprobe for dual-modal breast cancer imaging in the second near-infrared window.

BACKGROUND: Fluorescence imaging as the beacon for optical navigation has wildly developed in preclinical studies due to its prominent advantages, including noninvasiveness and superior temporal resolution. However, the traditional optical methods based on ultraviolet (UV, 200-400 nm) and visible light (Vis, 400-650 nm) limited by their low penetration, signal-to-noise ratio, and high background auto-fluorescence interference. Therefore, the development of near-infrared-II (NIR-II 1000-1700 nm) nanoprobe attracted significant attentions toward in vivo imaging. Regrettably, most of the NIR-II fluorescence probes, especially for inorganic NPs, were hardly excreted from the reticuloendothelial system (RES), yielding the anonymous long-term circulatory safety issue. RESULTS: Here, we develop a facile strategy for the fabrication of Nd3+-doped rare-earth core-shell nanoparticles (Nd-RENPs), NaGdF4:5%Nd@NaLuF4, with strong emission in the NIR-II window. What's more, the Nd-RENPs could be quickly eliminated from the hepatobiliary pathway, reducing the potential risk with the long-term retention in the RES. Further, the Nd-RENPs are successfully utilized for NIR-II in vivo imaging and magnetic resonance imaging (MRI) contrast agents, enabling the precise detection of breast cancer. CONCLUSIONS: The rationally designed Nd-RENPs nanoprobes manifest rapid-clearance property revealing the potential application toward the noninvasive preoperative imaging of tumor lesions and real-time intra-operative supervision.

Hydropenia may accelerates the progression of orthotopic bladder cancer induced by N-methyl-N-nitrosourea by increasing the expression levels of AQP1, AQP3, and AQP4.

BACKGROUND: Increasing evidence has demonstrated aquaporins (AQPs) to be critical players in carcinogenesis. Here, we aimed to explore the role of hydropenia in the progression of bladder cancer (BCa), as well as to assess the expression of AQP1, AQP3, and AQP4 in bladder tissues from hydropenic and N-methyl-N-nitrosourea (MNU)-treated rats. METHODS: An orthotopic BCa model was induced by administering Sprague Dawley rats with MNU. A hydropenic rat model was established by administrating rats with 2/3 of the amount of water given to the control group. At week 8, the rats were sacrificed and their bladder tissues were collected. Then, pathological alterations in the rat bladders were assessed by hematoxylin and eosin staining. The RNA and protein expression levels of AQP1, AQP3, and AQP4 were determined by using qRT-PCR and western blot assays. RESULTS: All of the rats (100%) administrated with MNU developed tumors, of which 5 were large (diameter, 0.5-1.0 cm), 10 were medium (diameter, 0.2-0.5 cm), and 5 were small (diameter, <0.2 cm) in size. The tumors were nodular and cauliflower shaped, with multiple satellite focus, and were accompanied by bleeding, ulcers, stones, and residual urine. Hematoxylin and eosin staining revealed that the bladder mucosa was incomplete, with a large amount of necrotic tissue and obvious leukocytic infiltration. The tumor volume in the MNU + hydropenia group was significantly larger than that in the MNU group. Noticeably, hydropenia exacerbated pathological changes induced by MNU administration. QRT-PCR and western blot analysis revealed that the MNU group, hydropenia group, and MNU + hydropenia group had significantly increased levels of AQP1, AQP3, and AQP4 compared to the control group, with the most dramatic increase seen in the MNU + hydropenia group. CONCLUSIONS: Hydropenia exacerbates pathological alterations induced by MNU in rats with orthotopic BCa by increasing the expression levels of AQP1, AQP3, and AQP4. This study reveals a possible mechanism of the occurrence of BCa.

Comparison of mini-percutaneous nephrolithotomy and retroperitoneal laparoscopic ureterolithotomy for treatment of impacted proximal ureteral stones greater than 15 mm.

BACKGROUND: The optimal treatment for large impacted proximal ureteral stones remains controversial. The aim of this study was to evaluate the efficacy, safety, and potential complications of mini-percutaneous nephrolithotomy (MPCNL) and retroperitoneal laparoscopic ureterolithotomy (RPLU) in the treatment of impacted proximal ureteral stones with size greater than 15 mm. METHODS: A total of 268 patients with impacted proximal ureteral stones greater than 15 mm who received MPCNL or RPLU procedures were enrolled consecutively between January 2014 and January 2019. Data on surgical outcomes and complications were collected and analyzed. RESULTS: Demographic and ureteral stone characteristics found between these two groups were not significantly different. The surgical success rate (139/142, 97.9% vs. 121/126, 96.0%, P = 0.595) and stone-free rate after 1 month (139/142, 97.9% vs. 119/126, 94.4%, P = 0.245) of RPLU group were marginally higher than that of the MPCNL group, but there was no significant difference. There was no significant difference in the drop of hemoglobin between the two groups (0.8 ±â€Š0.6 vs. 0.4 ±â€Š0. 2 g/dL, P = 0.621). The mean operative time (68.2 ±â€Š12.5 vs. 87.2 ±â€Š16.8 min, P = 0.041), post-operative analgesics usage (2/121, 1.7% vs. 13/139, 9.4%, P = 0.017), length of hospital stay after surgery (2.2 ±â€Š0.6 vs. 4.8 ±â€Š0.9 days, P < 0.001), double J stent time (3.2 ±â€Š0.5 vs. 3.9 ±â€Š0.8 days, P = 0.027), time of catheterization (1.1 ±â€Š0.3 vs. 3.5 ±â€Š0.5 days, P < 0.001), and time of drainage tube (2.3 ±â€Š0.3 vs. 4.6 ±â€Š0.6 days, P < 0.001) of MPCNL group were significantly shorter than that of the RPLU group. The complication rate was similar between the two groups (20/121, 16.5% vs. 31/139, 22.3%, P = 0.242). CONCLUSIONS: MPCNL and RPLU have similar surgical success and stone clearance in treating impacted proximal ureteral stones greater than 15 mm, while patients undergoing MPCNL had a lower post-operative pain rate and a faster recovery.

Needle Adjustment Free (NAF) running suture technique (PAN suture) in laparoscopic partial nephrectomy.

BACKGROUND: It is proposed a new running suture technique called Needle Adjustment Free (NAF) technique, or PAN suture. The efficiency and the safety were evaluated in laparoscopic partial nephrectomy. METHODS: This new running suture technique avoids the Needle Adjustment method used in traditional techniques. The new continuous suture technique (11 patients) was compared with the traditional continuous suture method (33 patients) used in both transperitoneal and retroperitoneal laparoscopic partial nephrectomy (LPN) in terms of suture time (ST), warm ischemia time (WIT), blood loss (BL), open conversion rate and post-op discharge time, post-op bleeding, post-op DVT, ΔGFR (affected side, 3 months post-op). Differences were considered significant when P < 0.05. RESULTS: ST in the PAN suture group was 30.37 ± 16.39 min, which was significant shorter (P = 0.0011) than in the traditional technique group which was 13.68 ± 3.33 min. WIT in the traditional technique group was 28.73 ± 7.89 min, while in the PAN suture group was 20.64 ± 5.04 min, P = 0.0028. The BL in entirety in the traditional technique group was 141.56 ± 155.23 mL, and in the PAN suture group was 43.18 ± 31.17 mL (P = 0.0017). BL in patients without massive bleeding in the traditional technique group was significantly greater than in the PAN suture group at 101.03 ± 68.73 mL versus 43.18 ± 31.17 mL (P = 0.0008). The open conversion rate was 0 % in both groups. There was no significant difference between the two groups in postoperative discharge time, post-op bleeding, post-op DVT, ΔGFR (affected side, 3 months post-op). CONCLUSIONS: The NAF running suture technique, or PAN suture, leading to less ST, WIT and BL, which was shown to be more effective and safer than the traditional technique used for LPN. A further expanded research with larger sample size is needed.

Identifying prognostic signatures in the microenvironment of prostate cancer.

BACKGROUND: An increasing number of studies has indicated that the tumor microenvironment (TME), an important component of tumor tissue, has clinicopathological significance in predicting disease outcome and therapeutic efficacy. However, little evidence in prostate cancer (PCa) is available. METHODS: The cohort of TCGA-PRAD (n=477) was used in this study. Based on the proportion of 22 types of immune cells calculated by CIBERSORT, the TME was classified by K-means clustering and differentially expressed genes (DEGs) were determined. The TMEscore was calculated based on cluster signature genes, which were obtained from DEGs by the random forest method, and the samples were classified into two subtypes. Analyses of somatic mutation and copy number variation (CNVs) were further conducted to identify the genetic characteristics of the two subtypes. Correlation analysis was performed to explore the correlation between TMEscore and the tumor response to immune checkpoint inhibitors (ICIs) as well as the prognosis of PCa. RESULTS: Based on the distribution of infiltrating immune cells in the TME, we constructed the TMEscore model and classified PCa samples into high and low TMEscore groups. Survival analysis indicated that the high TMEscore group had significantly better survival outcome than the low TMEscore group. Correlation analysis showed a significantly positive correlation between TMEscore and the known prognostic factors of PCa. CONCLUSIONS: Our study indicates that the TMEscore could be a potential prognostic biomarker in PCa. A comprehensive description of the characteristics of TME may help predict the response to therapies and provide new treatment strategies for PCa patients.

Factors Influencing Surgical Outcome in Retrograde Management of Parapelvic Renal Cysts.

Objective: To investigate the outcomes of retrograde flexible ureteroscopy in managing parapelvic renal cysts and speculate the factors affecting therapeutic efficacy. Methods: Thirty-eight patients with parapelvic renal cysts were recruited and underwent retrograde flexible ureteroscopy using holmium laser. Parapelvic cysts were divided into peripheral type and central type based on the position of cyst convex to the perirenal tissue. Feasibility and safety were retrospectively evaluated, and cases were analyzed to detect their distinctive characteristics. Independent-sample t-test and chi-square test were undertaken for continuous variables and categorical variables, respectively. Results: Radiologic evidence of success was achieved in 31 (81.58%) cases after a mean follow-up of 14.4 months (range 6-26 months). No significant perioperative complications were identified. There were seven cases with features of peripherally located parapelvic cyst. Four cysts shown as irregular protrusion were unable reduce to less half of previous size. Reductions were recorded in the other three patients with spherically peripheral protrusion. There was significant difference between these two types (p = 0.029). Among the 31 patients with centrally located parapelvic cyst, 28 of these have simple cysts that achieved radiologic success and 3 of the 31 patients were identified as failed cases indicated by renal pelvis enveloped by cyst on radiologic investigation. The success rate of simple cysts was significantly higher than that of the later type (p < 0.001). Conclusion: The location and shape of parapelvic cyst may play a critical role in the radiologic outcome of internal incision and patients with simple central or spherical peripheral cyst may benefit more from retrograde flexible ureteroscopy combined with laser incision.

Self-Assembled Hybrid Nanocomposites for Multimodal Imaging-Guided Photothermal Therapy of Lymph Node Metastasis.

Multimodal imaging-guided therapy holds great potential for precise theranostics of cancer metastasis. However, imaging agents enabling the convergence of complementary modalities with therapeutic functions to achieve perfect theranostics have been less exploited. This study reports the construction of a multifunctional nanoagent (FIP-99mTc) that comprises Fe3O4 for magnetic resonance imaging, radioactive 99mTc for single-photon-emission computed tomography, and IR-1061 to serve for the second near-infrared fluorescence imaging, photoacoustic imaging, and photothermal therapy treatment of cancer metastasis. The nanoagent possessed superior multimodal imaging capability with high sensitivity and resolution attributing to the complement of all the imaging modalities. Moreover, the nanoagent showed ideal photothermal conversion ability to effectively kill tumor cells at low concentration and power laser irradiation. In the in vivo study, FIP-99mTc confirmed the fast accumulation and clear delineation of metastatic lymph nodes within 1 h after administration. Attributing to the efficient uptake and photothermal conversion, FIP-99mTc could raise the temperature of metastatic lymph nodes to 54 °C within 10 min laser irradiation, so as to facilitate tumor cell ablation. More importantly, FIP-99mTc not only played an active role in suppressing cancer growth in metastatic lymph nodes with high efficiency but also could effectively prevent further lung metastasis after resection of the primary tumor. This study proposes a simple but effective theranostic approach toward lymph node metastasis.

Beforehand transection and suturing (BTS) of the dorsal vascular complex: a novel technique in laparoscopic radical prostatectomy.

BACKGROUND: Beforehand transection and suturing (BTS) of the dorsal vascular complex (DVC), a novel technique in non-neurovascular bundle sparing (NVB-sparing) extraperitoneal laparoscopic radical prostatectomy (eLRP), had been proposed; this study aimed to evaluate this technique in clinical laparoscopic procedures. METHODS: Using this new technique, the DVC was transected and sutured after dissection of the pelvic fascia and before dissection of the prostate, especially before ligation of the bilateral prostatic pedicles. This study retrospectively analyzed the data of 90 non NVB-sparing eLRP patients [traditional technique (n=60) and BTS technique (n=30)]. RESULTS: The surgical time in the BTS technique group was 121.73±24.53 min, which was significantly shorter (P=0.0015) than the traditional technique group (144.12±39.68 min). The calculated blood loss in the traditional technique group was 388.45±232.78 mL, and 264.16±130.70 mL in the BTS technique group (P=0.0016). The estimated blood loss in the traditional technique group was 350.34±311.80 mL, which was significantly greater than the BTS technique group (250.33±145.31 mL, P=0.0422). The transfusion rate in the traditional technique group was significantly greater than the BTS technique group (15.00% vs. 0.00%; P=0.0266). The biochemical recurrence rate in traditional technique group was 48.33%, which was higher than in the BTS group (30.00%) (P=0.0465). There was no significant difference between the 2 groups with respect to the pre-operative hemoglobin (Hb) concentration, pre-operative hematocrit (HCT), post-operative Hb concentration, post-operative HCT, ΔHCT, pre-operative blood volume, rectal perforation, open conversion, apical capsule residue, false suture, post-operative bleeding, urinary leakage, re-operation, surgical site infection, post-operative stay, and emission time of urinary incontinence. CONCLUSIONS: In managing the relationship between the DVC and prostate in patients undergoing non NVB-sparing eLRP, the BTS technique was shown to be more effective and safer than the traditional technique.

[Establishment of reference ranges of sex hormones for healthy children in Shenzhen, China based on chemiluminescence].

OBJECTIVE: To study the reference ranges of six sex hormones, i.e., luteinizing hormone, follicle-stimulating hormone, progesterone, prolactin, estradiol, and testosterone, for healthy children aged 0-18 years in Shenzhen, China. METHODS: Stratified cluster sampling was performed to select 2 178 healthy children aged 0-18 years in the districts of Futian, Luohu, Nanshan, Bao'an, and Longgang in Shenzhen between September 2015 and September 2016. There were 1 219 boys and 959 girls, including 81 neonates, 335 infants, 346 young children, 469 preschool children, 419 school-aged children, and 528 adolescents. The American Beckman DXI800 chemiluminescence meter was used to measure the levels of luteinizing hormone, follicle-stimulating hormone, progesterone, prolactin, estradiol, and testosterone. RESULTS: There were significant differences in the levels of luteinizing hormone, follicle-stimulating hormone, progesterone, prolactin, estradiol, and testosterone between different age groups (P<0.05). There were also significant differences in the levels of these sex hormones between boys and girls in the same age group (P<0.05). The reference ranges of six sex hormones were established for healthy children aged 0-18 years in Shenzhen based on the levels of these hormones in different age groups. CONCLUSIONS: There are significant differences in sex hormones between different age groups or sex groups. The reference ranges of six sex hormones established for different sexes or ages have great significance in the diagnosis and treatment of endocrine diseases in children.

TIGAR knockdown radiosensitizes TrxR1-overexpressing glioma in vitro and in vivo via inhibiting Trx1 nuclear transport.

The up-regulation of thioredoxin reductase-1 (TrxR1) is detected in more than half of gliomas, which is significantly associated with increased malignancy grade and recurrence rate. The biological functions of NADPH-dependent TrxR1 are mainly associated with reduced thioredoxin-1 (Trx1) which plays critical roles in cellular redox signaling and tumour radio-resistance. Our previous work has proved that TP53 induced glycolysis and apoptosis regulator (TIGAR) knockdown could notably radiosensitize glioma cells. However, whether TrxR1-overexpressing glioma cells could be re-radiosensitized by TIGAR silence is still far from clear. In the present study, TrxR1 was stably over-expressed in U-87MG and T98G glioma cells. Both in vitro and in vivo data demonstrated that the radiosensitivity of glioma cells was considerably diminished by TrxR1 overexpression. TIGAR abrogation was able to radiosensitize TrxR1-overexpressing gliomas by inhibiting IR-induced Trx1 nuclear transport. Post-radiotherapy, TIGAR low-expression predicted significant longer survival time for animals suffering from TrxR1-overexpessing xenografts, which suggested that TIGAR abrogation might be a promising strategy for radiosensitizing TrxR1-overexpressing glial tumours.

[Analysis of clinical characteristics of hypoxic hepatitis in children].

OBJECTIVE: To explore the etiology and clinical characteristics of hypoxic hepatitis (HH) in children. METHOD: Clinical data of 7 patients with HH in Shenzhen Children's Hospital from January 2011 to March 2014 were retrospectively reviewed. RESULT: Seven cases diagnosed as HH, age from 4 months to 11 years, were admitted to pediatric intensive care unit (PICU), and accounted for 0.32% of patients in PICU during the same period. The primary causes of HH were respiratory failure and cardiac shock caused by severe hand-foot-and-mouth disease, fulminant myocarditis, infant muggy syndrome . Serologic tests for hepatitis B virus, hepatitis C virus, as well as serum antibody and DNA for Epstein-Barr virus and cytomegalovirus were all negative. There was an increase of alanine aminotransferase (ALT) (≥20 time supper limit of normal (ULN), the highest ALT was more than 130 times ULN in all the patients, which was decreased to 2 times ULN from peak within 10 days. There was a significant relationship between ALT and aspartate aminotransferase(AST)in 3 cases(r=1.000, 1.000, and 0.833, respectively, P<0.05), ALT and lactate dehydrogenase (LDH)in 2 cases(r=1.000 and 0.886, respectively, P<0.05), ALT and blood urea nitrogen(BUN)in 1 case(r=1.000, P<0.05), and ALT and creatine kinase(CK)in 1 case(r=0.964, P<0.05). The ALT, AST and LDH returned to normal soon after the primary diseases were controlled. CONCLUSION: Severe heart failure, hypoxemia, shock, etc. are the leading primary diseases causing HH. The sharp increase in ALT, AST and LDH is the typical laboratory manifestion in HH after the onset, which may decline to normal shortly after the treatment, sometimes complicated with reversible change in BUN or CK.

Pathologic analysis, diagnosis and treatment of adrenal myelolipoma.

With the widespread use of non-invasive imaging modalities, the incidental detection of myelolipoma has become more common. We analyze the clinical data of 4 typical cases of unilateral adrenal myelolipomas, including 1 hormonally inactive patient and 3 hormonally active cases with hyperaldosteronemia, hypercortisolism and hyperandrogenemia. Obvious differences were found in the pathological sections. To our knowledge, this is the first article discussing the etiology of adrenal myelolipoma from pathologic analyses.

Metachronous bilateral adrenocortical functional adenomas causing adrenocorticotropic hormone-independent Cushing's syndrome.

Metachronous bilateral adrenocortical adenomas causing adrenocorticotropic hormone-independent Cushing's syndrome is a surgical case that scarcely occurs. A 38-year-old woman diagnosed with bilateral adrenocortical adenomas one by one in 2006 and 2013. Both of the two adenomas were functional and caused typical Cushingoid symptoms. The patient underwent a laparoscopic adrenolectomy in 2006 and a laparoscopic adrenocortical adenoma resection in 2013. After surgery, the symptoms of Cushing's syndrome withdrew and the woman started steroid replacement therapy as a following treatment.

Clear cell changes in mucinous tubular and spindle cell carcinoma: cytoplasmic pallor/clearing within tubules, vacuoles or hybrid conventional clear cell carcinoma of kidney?

Mucinous tubular and spindle cell carcinoma (MTSCC) is a rare and recently recognized subtype of renal cell carcinoma (RCC). Apart from the classic morphology comprising conventional three components, there exist a large number of non-classic morphological variants of MTSCC, which make it necessity to differentiate from other RCC. Herein, we report two non-classic morphological variants of MTSCC. Case 1, a 85 years old man, showed numerous vacuoles among inherent components and cytoplasmic pallor/clearing within tubules mimicking conventional clear cell RCC with a 8.5 years follow-up, while Case 2 indicated a "mucin-poor" MTSCC associated with simultaneous conventional clear cell RCC at her age of 73 years. Until now Case 1 carries the longest disease-free survival reported in literature since MTSCC was defined and ranks the oldest since reported in literature, while Case 2 is the first report of "mucin-poor" MTSCC associated with simultaneous conventional clear cell RCC. Now, since no biomarkers or imagining tools but pathological examination can confirm the diagnosis of MTSCC, the management is always following the guideline of RCC in clinical practice. Generally, most reports consider it as a good prognosis disease, but sarcomatoid variant, even classic subtype can progress rapidly to life-threatening disease.