CRISPR genome-wide screening identifies PAK1 as a critical driver of ARSI cross-resistance in prostate cancer progression.

Next-generation androgen receptor signaling inhibitors (ARSIs), such as enzalutamide (Enza) and darolutamide (Daro), are initially effective for the treatment of advanced prostate cancer (PCa) and castration-resistant prostate cancer (CRPC). However, patients often relapse and develop cross-resistance, which consequently makes drug resistance an inevitable cause of CRPC-related mortality. By conducting a comprehensive analysis of GEO datasets, CRISPR genome-wide screening results, ATAC-seq data, and RNA-seq data, we systemically identified PAK1 as a significant contributor to ARSI cross-resistance due to the activation of the PAK1/RELA/hnRNPA1/AR-V7 axis. Inhibition of PAK1 followed by suppression of NF-κB pathways and AR-V7 expression effectively overcomes ARSI cross-resistance. Our findings indicate that PAK1 represents a promising therapeutic target gene for the treatment of ARSI cross-resistant PCa patients in the clinic. STATEMENT OF SIGNIFICANCE: PAK1 drives ARSI cross-resistance in prostate cancer progression.

Tumor-associated macrophages promote migration and invasion via modulating IL-6/STAT3 signaling in renal cell carcinoma.

Tumor-associated macrophages (TAMs) promote tumor cell growth and metastasis in various human cancers. However, the role of TAMs in renal cell carcinoma (RCC) is rarely investigated. Herein, we observed that the infiltration of TAMs was obviously elevated in RCC tumor tissues, high infiltration of TAMs was closely associated with tumor progression and poor prognosis in RCC patients. In vitro assays further indicated that the conditioned medium of TAMs (TAMs CM) facilitated migration, invasion, and epithelial-mesenchymal transition (EMT) in RCC cells. Moreover, we found that IL-6 was involved in the functions of TAMs in RCC; IL-6 neutralizing antibody (IL-6NA) partly abolished the effect of TAMs on RCC cells. In addition, we demonstrated that TAMs might exert their roles by activating STAT3 signaling in RCC, and IL-6 was responsible for TAMs-induced STAT3 signaling activation. In conclusion, our results revealed that high infiltration of TAMs may promote RCC cells migration, invasion, and EMT via modulating IL-6/STAT3 signaling, further suggesting a potential of novel treatment strategies targeting TAMs or IL-6 for metastatic RCC.

Modified Prostate Health Index Density Significantly Improves Clinically Significant Prostate Cancer (csPCa) Detection.

Background: Early screening of clinically significant prostate cancer (csPCa) may offer opportunities in revolutionizing the survival benefits of this lethal disease. We sought to introduce a modified prostate health index density (mPHI) model using imaging indicators and to compare its diagnostic performance for early detection of occult onset csPCa within the prostate-specific antigen (PSA) gray zone with that of PHI and PHID. Methods and Participation: Between August 2020 and January 2022, a training cohort of 278 patients (total PSA 4.0-10.0 ng/ml) who were scheduled for a prostate biopsy were prospectively recruited. PHI and PHID were compared with mPHI ( LD TRD × APD × TPV × PHI ) for the diagnosis performance in identifying csPCa. Pathology outcomes from systematic prostate biopsies were considered the gold standard. Results: This model was tested in a training cohort consisting of 73 csPCa, 14 non-clinically significant prostate cancer(non-csPCa), and 191 benign prostatic hyperplasia (BPH) samples. In the univariate analysis for the PSA gray zone cohort, for overall PCa, the AUC of mPHI (0.856) was higher than PHI (0.774) and PHID (0.835). For csPCa, the AUC of mPHI (0.859) also surpassed PHI (0.787) and PHID (0.825). For detection of csPCa, compared with lower specificities from PHI and PHID, mPHI performed the highest specificity (76.5%), by sparing 60.0% of unnecessary biopsies at the cost of missing 11 cases of csPCa. The mPHI outperformed PHI and PHID for overall PCa detection. In terms of csPCa, mPHI exceeds diagnostic performance with a better net benefit in decision curve analysis (DCA) compared with PHI or PHID. Conclusions: We have developed a modified PHI density (mPHI) model that can sensitively distinguish early-stage csPCa patients within the PSA gray zone. Clinical Trial Registration: ClinicalTrials.gov, NCT04251546.

In Vivo Flow Cytometry.

In vivo flow cytometry (IVFC) was first designed to detect circulating cells in a mouse ear. It allows real-time monitoring of cells in peripheral blood with no need to draw blood. The IVFC field has made great progress during the last decade with the development of fluorescence, photoacoustic, and multiphoton microscopy. Moreover, the application of IVFC is no longer restricted to circulating cells. IVFC based on fluorescence and photoacoustic are most widely applied in biomedical research. Methods based on fluorescence are often used for object monitoring in superficial vessels, while methods based on photoacoustics have an advantage of label-free monitoring in deep vessels. In this chapter, we introduce technical points and key applications of IVFC. We focus on the principles, labeling strategies, sensitivity, and biomedical applications of the technology. In addition, we summarize this chapter and discuss important research directions of IVFC in the future.

Characteristics and prognostic value of potential dependency genes in clear cell renal cell carcinoma based on a large-scale CRISPR-Cas9 and RNAi screening database DepMap.

BACKGROUND: Large-scale loss-of-function screening database such as Cancer Dependency Map (Depmap) provide abundant resources. Investigation of these potential dependency genes from human cancer cell lines in the real-world patients cohort would evaluate their prognostic value thus facilitate their clinical application and guide drug development. METHODS: A few genes were selected from top clear cell renal cell carcinoma (ccRCC) lineage preferential dependency candidates from Depmap. Their characteristic including expression levels both in normal and tumor tissues and correlations with methylation or copy number, genetic alterations, functional enrichment, immune-associated interactions, prognostic value were evaluated in KIRC cohort from TCGA, GTEx, and multiple other open databases and platforms. RESULTS: 16 genes were collected from 106 ccRCC preferential candidates and further analyzed including B4GALT4, BCL2L1, CDH2, COPG1, CRB3, FERMT2, GET4, GPX4, HNF1B, ITGAV, MDM2, NFE2L2, PAX8, RUVBL1, TFRC, and TNFSF10. The normalized gene effect scores of these genes varied from different ccRCC cell lines and principal component analysis (PCA) showed their tissue specificity expression profiles. Genetic alteration rates of them were low to moderate (0.7%-13%) in KIRC cohort. CDH2, MDM2, TNFSF10 showed a statistically significant higher level in tumors than normal tissues while PAX8 and FERMT2 were significantly downregulated. Moderate positive or negative correlations were observed in several genes between their expression and relative gene copy number or methylation levels, respectively. Based on the multivariable COX regression model adjusted by critical clinical variables revealed the expression of GET4 (p=0.002, HR=1.023 95%CI 1.009-1.038) and CRB3 (p<0.001, HR=0.969 95%CI 0.960-0.980) were independent predictive factors for overall survival in KIRC cohort. CONCLUSIONS: A dependency gene validated in cell lines didn't directly represent its role in corresponding patients with same histological type and their prognostic value might be determined by multiple factors including dependency driven types, genetic alteration rates and expression levels. GET4 and CRB3 were the independent prognostic factors for ccRCC patients. CRB3 seemed like a potential broad tumor suppressor gene while GET4 might be a ccRCC preferential dependency gene with a ligandable structure.

Detection and Classification of Multi-Type Cells by Using Confocal Raman Spectroscopy.

Tumor cells circulating in the peripheral blood are the prime cause of cancer metastasis and death, thus the identification and discrimination of these rare cells are crucial in the diagnostic of cancer. As a label-free detection method without invasion, Raman spectroscopy has already been indicated as a promising method for cell identification. This study uses a confocal Raman spectrometer with 532 nm laser excitation to obtain the Raman spectrum of living cells from the kidney, liver, lung, skin, and breast. Multivariate statistical methods are applied to classify the Raman spectra of these cells. The results validate that these cells can be distinguished from each other. Among the models built to predict unknown cell types, the quadratic discriminant analysis model had the highest accuracy. The demonstrated analysis model, based on the Raman spectrum of cells, is propitious and has great potential in the field of biomedical for classifying circulating tumor cells in the future.

Resveratrol could attenuate prostatic inflammation in rats with Oestradiol-induced chronic prostatitis.

To evaluate the effect of resveratrol in rats with chronic prostatitis, 24 rats were randomly divided into the negative control, vehicle-treated and resveratrol groups. The rats in the vehicle-treated group and the resveratrol group were injected subcutaneously with 17-ß-oestradiol (0.25 mg/kg) daily for 6 weeks while the rats in the control group were injected with equivalent normal saline. From the 45th day, the rats in the resveratrol group were given resveratrol (10 mg/kg) by gavage per day while the rest rats were given normal saline. After 55 days, all the rats were sacrificed and the prostatic tissue was removed. Morphological changes were examined by light microscope after H&E staining. The expressions of IL-6, IL-8 and TNF-α were determined through ELISA and immunohistochemical staining. As a result, significant inflammatory cell infiltration and fibroblastic hyperplasia were observed in prostatic stroma in the vehicle-treated group compared with the negative control group, as well as the high expression of IL-6, IL-8 and TNF-α. After resveratrol treatment, inflammatory cell infiltration and fibroblastic hyperplasia were shown prominently reduced. Meanwhile, the expression of IL-6, IL-8 and TNF-α was significantly suppressed. For conclusion, resveratrol could attenuate the prostatic inflammation and downregulate the expression of IL-6, IL-8 and TNF-α in rat with oestradiol-induced chronic prostatitis.

A novel definition of microvessel density in renal cell carcinoma: Angiogenesis plus vasculogenic mimicry.

The present study proposed the novel concept of total microvessel density (TMVD), which is the combination of the MVD and the vasculogenic mimicry (VM) status, and evaluated its clinical significance in patients with renal cell carcinoma (RCC). For that purpose, tumor samples from 183 patients with primary RCC were examined by CD34 single or periodic acid Schiff (PAS)/CD34 dual histology staining. MVD and VM were determined according to previous literature. Clinical information (tumor stage and grade, and duration of survival) was retrieved and analyzed. Survival information and VM-associated gene expression data of patients with RCC were also retrieved from The Cancer Genome Atlas (TCGA) database and the clinical significance of each individual gene was analyzed. The results indicated that MVD exhibited obvious differences among patients with RCC; however, it was not correlated with the stage/grade or length of survival in patients with RCC. In total, 81 patients (44.3%) were CD34(-)/PAS(+) and defined as VM(+), and they had a significantly shorter survival compared with that of VM(-) patients (P=0.0002). VM was not associated with MVD. TMVD was able to distinguish between patients with high and low MVD in terms of survival, thus TMVD was better compared with MVD alone at distinguishing between patients with different survival prognoses. TCGA data analysis revealed that among the VM-associated genes, nodal growth differentiation factor, caspase-3, matrix metalloproteinase-9 and galectin-3 had a statistically significant impact on the overall/disease-free survival of patients with RCC. In conclusion, the TMVD concept may be more appropriate and sensitive compared with the MVD or VM alone in predicting tumor aggressiveness and patient survival, particularly in RCC, which is a highly vascularized, VM-rich neoplasm, and certain VM formation-associated genes are negatively associated with the survival of patients with RCC.

Compare the Clinical Application of Ureteroscopic Occluder and Stone Retrieval Basket During Holmium Laser Treatment for Upper Ureteral Calculi.

PURPOSE: To explore the clinical efficacy of ureteroscopic occluder and stone retrieval basket combined with holmium laser in the treatment of upper ureteral calculi. MATERIALS AND METHODS: This retrospective study included 103 patients treated with ureteroscopic holmium laser lithotripsy for upper ureteral stones. Patients were divided into two groups based on the device applied during lithotripsy: group 1 for the occluders (52 cases), and group 2 for the stone retrieval baskets (51 cases). The stone upward migration rate, stone-free rate, and complication rate during or after surgery were compared. RESULTS: The operation time was 45 ± 7 min in the occluder group and 43 ± 5 min in the basket group (P = .111). There was no significant difference between the stone retropulsion rate (13% vs. 16%, P = .787). The successful one-time stone-free rate was 92% vs. 94% (P = .999) respectively. Furthermore, there was no significant difference in the hospitalization time (P = .581) and postoperative complication rate (P = .715) between 2 groups. CONCLUSION: The treatment of upper ureteral calculi with ureteroscopic occluder and stone retrieval basket combined with holmium laser lithotripsy can both effectively prevent intraoperative stone retropulsion, improve the success rate of one-time lithotrips. The occluder was more cost-effective than the stone retrieval basket, yet it was a more desired choice for over dilated ureters.

Visualizing Interactions of Circulating Tumor Cell and Dendritic Cell in the Blood Circulation Using In Vivo Imaging Flow Cytometry.

OBJECTIVE: Visualizing cell interactions in blood circulation is of great importance in studies of anticancer immunotherapy or drugs. However, the lack of a suitable imaging system hampers progress in this field. METHODS: In this work, we built a dual-channel in vivo imaging flow cytometer to visualize the interactions of circulating tumor cells (CTCs) and dendritic cells (DCs) simultaneously in the bloodstream. Two artificial neural networks were trained to identify blood vessels and cells in the acquired images. RESULTS AND CONCLUSION: Using this technique, single CTCs and CTC clusters were readily distinguished by their morphology. Interactions of CTCs and DCs were identified, while their moving velocities were analyzed. The CTC-DC clusters moved at a slower velocity than that of single CTCs or DCs. This may provide new insights into tumor metastasis and blood rheology. SIGNIFICANCE: This in vivo imaging flow cytometry system holds great potential for assessing the efficiency of targeting CTCs with anticancer immune cells or drugs.

Monitoring circulating prostate cancer cells by in vivo flow cytometry assesses androgen deprivation therapy on metastasis.

It remains controversial whether surgical castration prolongs survival rate and improves therapy prospects in patients suffering from prostate cancer. We used PC3 cell line to establish prostate tumor models. In vivo flow cytometry and ultrasonic imaging were used to monitor the process of prostate cancer growth, development and metastasis. We found out that the number of circulating tumor cells (CTCs) in orthotopic tumor model was higher than that in subcutaneous tumor model. The CTC number in orthotopic tumor model was due to burst growth, while CTC number in subcutaneous tumor model showed a gradual increase with tumor size. After androgen deprivation therapy (ADT) through testicular extraction, we constructed GFP-PC3 subcutaneous tumor models and orthotopic tumor models. We found dramatically decreased CTC number, relieved symptoms caused by the tumor, and significantly prolonged survival time after testicular extraction in orthotopically transplanted prostate tumor model, while the carcinogenesis process and metastases were little influenced by ADT in subcutaneous tumor model. ADT treatment can restrict tumor growth, decrease the CTC number significantly and inhibit distant invasion through inhibition of tumor proliferation and tumor angiogenesis in orthotopical prostate tumor model. © 2018 International Society for Advancement of Cytometry.

Systemically Infused Mesenchymal Stem Cells Show Different Homing Profiles in Healthy and Tumor Mouse Models.

Bone marrow-derived mesenchymal stem cells (MSCs) can localize in injured, inflamed, and cancerous tissues after systemic infusion. However, the dynamic homing profile of MSCs in the peripheral blood is not well characterized. Here, using in vivo flow cytometry to noninvasively monitor the dynamics of fluorescence-labeled cells, we found different clearance kinetics of systemically infused MSCs between healthy and tumor mouse models. The circulation times of MSCs in healthy mice and mice with subcutaneous tumors, orthotopically transplanted liver tumors, or metastatic lung tumors were 30, 24, 18, and 12 hours, respectively, suggesting that MSCs actively home to tumor environments. MSCs infiltrated into hepatocellular carcinoma (HCC) sites and preferentially engrafted to micrometastatic regions both in vivo and in vitro. The expression of epidermal growth factor, CXCL9, CCL25, and matrix metalloproteinases-9 by HCC cells differed between primary tumor sites and metastatic regions. By characterizing the homing profiles of systemically perfused MSCs under physiological and cancerous conditions, these findings increase our understanding of the migration of MSCs from the circulation to tumor sites and constitute a basis for developing MSC-based anti-cancer therapeutic strategies. Stem Cells Translational Medicine 2017;6:1120-1131.

A possible relationship between serum sex hormones and benign prostatic hyperplasia/lower urinary tract symptoms in men who underwent transurethral prostate resection.

In this study, we examined the relationship between sex hormone levels and lower urinary tract symptoms (LUTS) in men with benign prostatic hyperplasia (BPH) who underwent transurethral surgery. The study was conducted in 158 patients who came to our hospital for surgery. Clinical conditions were assessed by body mass index (BMI), digital rectal examination, International Prostate Symptom Score (IPSS) and transrectal ultrasound (TRUS). The levels of sex hormones (including total testosterone (TT), estradiol (E 2 ), progesterone (P), luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin (PRL)) and prostate-specific antigen (PSA) were reviewed. Correlations were determined through statistical analysis. The mean age was 72.06 ± 8.68 years. The total IPSS was significantly associated with the TT level (r = -0.21, P= 0.01). Other sex hormone levels were not correlated with total IPSS. However, some ratios such as E 2 / TT (r = 0.23, P= 0.00) and FSH/LH (r = -0.17, P = 0.04) were associated with total IPSS. Further analysis showed that the nocturia was associated with age (r = 0.16, P= 0.04), BMI (r = 0.21, P = 0.01), and TT (r = -0.19, P= 0.02). Moreover, we divided the patients into two subgroups based on IPSS severity (<20 or ≥20). The mean TT level was in the normal range, but it was significantly related to the presence of severe LUTS. In summary, our study has shown that the severity of LUTS is associated with TT, E 2 / TT and FSH/LH in men who underwent prostate surgery. Increasing nocturia was observed in lower testosterone patients. Additional larger studies are needed to elucidate the potential mechanisms.

Long non­coding RNA­GAS5 acts as a tumor suppressor in bladder transitional cell carcinoma via regulation of chemokine (C­C motif) ligand 1 expression.

Long non­coding RNAs (lncRNAs) have important roles in diverse biological processes, including transcriptional regulation, cell growth and tumorigenesis. The present study aimed to investigate whether lncRNA­growth arrest­specific (GAS)5 regulated bladder cancer progression via regulation of chemokine (C­C) ligand (CCL)1 expression. The viability of BLX bladder cancer cells was detected using a Cell Counting kit­8 assay, and cell apoptosis was assessed by annexin V­propidium iodide double­staining. The expression levels of specific genes and proteins were analyzed by reverse transcription­quantitative polymerase chain reaction and western blotting, respectively. In addition, cells were transfected with small interfering (si)RNAs or recombinant GAS5 in order to silence or overexpress GAS5, respectively. The results of the present study demonstrated that knockdown of GAS5 expression promoted bladder cancer cell proliferation, whereas overexpression of GAS5 suppressed cell proliferation. Furthermore, knockdown of GAS5 resulted in an increased percentage of cells in S and G2 phase, and a decreased percentage of cells in G1 phase. In addition, the present study performed a hierarchical cluster analysis of differentially expressed lncRNAs in bladder cancer cells and detected that CCL1 overexpression resulted in an upregulation of GAS5, which may improve the ability of cells to regulate a stress response in vitro. Furthermore, knockdown of GAS5 expression increased the mRNA and protein expression of CCL1 in bladder cancer cells. Gain­of­function and loss­of­function studies demonstrated that GAS5 was able to inhibit bladder cancer cell proliferation, at least in part, by suppressing the expression of CCL1. The results of the present study demonstrated that GAS5 was able to suppress bladder cancer cell proliferation, at least partially, by suppressing the expression of CCL1. The results of the present study may provide a basis for developing novel effective treatment strategies against bladder cancer.

CXCR4-mediated Stat3 activation is essential for CXCL12-induced cell invasion in bladder cancer.

CXCL12/CXCR4 signaling plays important roles in tumor cell metastasis in many types of cancers, and CXCR4 is the key regulator of cell motility in bladder cancer. Emerging evidence suggests that transcription-3 (Stat3) activation is associated with bladder cancer cell growth and survival, while the relationship between CXCL12/CXCR4 signal and Stat3 activation remains unclear. In this study, expression analysis of bladder cancer and adjacent normal tissues showed that higher CXCR4 expression was associated with Stat3 phosphorylation. CXCR4 knockdown in bladder cancer T24 cells impaired CXCL12-induced cell invasion and Stat3 activation. Furthermore, blocking Stat3 activity with the chemical inhibitor Stattic inhibited CXCL12-triggered Stat3 phosphorylation and cell invasion in T24 cells, suggesting that Stat3 activation is required for CXCL12 function in the mobility of bladder cancer. Taken together, CXCR4 is necessary for CXCL12 signal transduction in bladder cancer, and CXCL12/CXCR4 promotes invasion of bladder cancer cells through activation of Stat3 transcriptional activity.

In vitro study on the binding of gemcitabine to bovine serum albumin.

In the present study, the interaction of gemcitabine and bovine serum albumin (BSA) has been characterized by spectroscopic methods (fluorescence, UV-vis, circular dichroism (CD) and Fourier transform infrared (FT-IR) spectroscopy) and molecular docking. Gemcitabine quenched BSA fluorescence in a static mode with binding constants of 6.61, 6.18, and 3.44 × 104M⁻¹ at 290, 300, and 310 K, respectively. Meanwhile, the number of binding site was found to be approximately 1 from fluorescence titration data. The calculated thermodynamic parameters represent a spontaneous process and electrostatic force dominated binding, which was confirmed by the docking study. Furthermore, the alterations of protein secondary structure in the presence of gemcitabine were assessed by CD UV-vis and FT-IR spectroscopy. Fluorescence resonance energy transfer (FRET) analysis proved high probability of energy transfer from Trp residue to the drug molecule.

Circulation times of prostate cancer and hepatocellular carcinoma cells by in vivo flow cytometry.

In metastasis, the cancer cells that travel through the body are capable of establishing new tumors in locations remote from the site of the original disease. To metastasize, a cancer cell must break away from its tumor and invade either the circulatory or lymphatic system, which will carry it to a new location, and establish itself in the new site. Once in the blood stream, the cancer cells now have access to every portion of the body. Here, we have used the "in vivo flow cytometer" to study if there is any relationship between metastatic potential and depletion kinetics of circulating tumor cells. The in vivo flow cytometer has the capability to detect and quantify continuously the number and flow characteristics of fluorescently labelled cells in vivo. We have improved the counting algorithm and measured the depletion kinetics of cancer cells with different metastatic potential. Interestingly, more invasive PC-3 prostate cancer cells are depleted faster from the circulation than LNCaP cells. In addition, we have measured the depletion kinetics of two related human hepatocellular carcinoma (liver cancer) cell lines, high-metastatic HCCLM3 cells, and low-metastatic HepG2 cells. More than 60% HCCLM3 cells are depleted within the first hour. Interestingly, the low-metastatic HepG2 cells possess noticeably slower depletion kinetics. In comparison, <40% HepG2 cells are depleted within the first hour. The differences in depletion kinetics might provide insights into early metastasis processes.

Management of renal cell carcinoma with tumor thrombus in renal vein and the inferior vena cava.

BACKGROUND: The article aims to sum up experience in the treatment of renal cell carcinoma complicated with tumor thrombus in renal vein and inferior vena cava. METHODS: A retrospective review was made on the diagnosis, treatment, and prognosis of 15 cases of renal carcinoma complicated with venous tumor thrombus from July 1994 to July 2006. RESULTS: The diagnosis of 93% (14/15) cases was confirmed by preoperative computed tomography or magnetic resonance imaging. Of the 15 cases, two had simple renal vein tumor thrombus of left kidney and 13 had inferior vena cava tumor thrombus; of the latter, nine were type I (pararenal type), three type II (subhepatic type), and one type III (intrahepatic type). Of the 12 patients who received surgical treatment, 11 had the renal tumors completely resected, the venous tumor thrombus removed, and lymph nodes cleared. Palliative excision was performed in one patient with a left kidney tumor because of adjacent adhesion. All the three patients who did not receive surgical treatment died, with a mean survival period of 7 months. Of the 12 surgical patients who received surgical treatment, three were lost during follow-up, and the other nine were followed up for 4-72 months; of these 9 patients, three (25%) survived tumor-free for more than 5 years, three for 1-3 years, and the other three died of metastasis within 1 year. CONCLUSION: Computerized tomography and magnetic resonance imaging are the best choice for noninvasive diagnosis of renal cell carcinoma complicated with inferior vena cava tumor thrombus. For patients without metastasis, radical resection of both the tumor and the thrombus often offers a relatively satisfactory outcome.

Percutaneous nephroscopic with holmium laser and ultrasound lithotripsy for complicated renal calculi.

The aim of this work is to validate the clinical efficacy of the high-power holmium:YAG laser with percutaneous nephrolithotripsy (PCNL) in combination with ultrasound lithotripsy for complicated renal calculi. From November 2006 to December 2007, 60 patients with complicated renal calculi were treated with PCNL, where an F24 standard renal access tract was established by percutaneous renal puncture under the guidance of B-mode ultrasound, and stones were fragmented and cleared by high-power holmium laser in combination with ultrasound under an F20.8 nephroscope. Of the 60 patients with complicated renal calculi, 20 were complete staghorn calculi and 30 were partial staghorn calculi, of which six patients were accompanied with renal insufficiency; two were solitary calculi, and eight were caliceal diverticular calculi. Calculi were removed by one attempt in 49 patients and by two attempts in 11 patients; through one tract in 50 patients and through two and three tracts in ten patients. The stone-free rate was 81.7%. No injury to the pleura and abdominal organs occurred during the intraoperative puncture. No postoperative blood transfusion was needed in any patient, nor did fever and secondary hemorrhage occur. The mean operation duration was 98 min (range, 60-150 min), and the mean lithotripsy time was 45 min (range, 30-85 min). Additional postoperative extracorporeal shock wave lithotripsy (ESWL) was performed on six patients. High-power holmium laser PCNL in combination with ultrasound lithotripsy is safe, effective, and minimally invasive, with a high stone-free rate, especially for complicated renal calculi.

[Suppression of invasion and angiogenesis in human prostate cancer PC-3 cells by adenovirus-mediated co-transfer of PTEN and P27].

OBJECTIVE: To investigate the effects of adenovirus-mediated PTEN and P27 on the invasion of PC-3 in vitro and angiogenesis, along with their synergy in the treatment of prostate cancer. METHODS: Recombinant adenovirus vectors of the human tumor suppressor genes PTEN and P27 were constructed. The replication-incompetent recombinant adenovirus was packaged and propagated in HEK293 cells. The viral titer was examined by plaque assay and the mRNA and protein expressions of PTEN and P27 in human prostate cancer cell line PC-3 infected with Ad-PTEN and Ad-P27 were determined by RT-PCR and Western blot respectively. The invasion of PC-3 cells in vitro was examined by Boyden chamber assay. MTT assay was used to testify the effect of supernatant from PC-3 infected with Ad-PTEN and Ad-P27 on the proliferation of endothelial cells ECV-304 and the CAM test was used to testify the effect of PTEN and P27 on angiogenesis. The difference between the combined therapy group and the single gene therapy group was also examined. RESULTS: The viral titers of Ad-PTEN and Ad-P27 were 1.8 x 10(7) pfu/ml and 1.2 x 10(9) pfu/ml respectively. Adenovirus infection verified that the mRNA and protein expression of PTEN and P27 were steady in human PC-3 cells. The invasion in vitro of PC-3 cells was significantly inhibited by infection with Ad-PTEN or/and Ad-P27. CAM and MTT assays of ECV-304 confirmed that the supernatant from PC-3 cells infected with Ad-PTEN or/and Ad-P27 could inhibit the angiogenesis effectively. There was a significant difference between the combined therapy group and the single gene therapy group. CONCLUSION: The combined gene therapy of Ad-PTEN and Ad-P27 plays a synergistic role in inhibiting the invasiveness of PC-3 cells and angiogenesis.