Publications by authors named "Hao-Cheng Lin"

27 Publications

  • Page 1 of 1

Room-Temperature Fabricated Multilevel Nonvolatile Lead-Free Cesium Halide Memristors for Reconfigurable In-Memory Computing.

ACS Nano 2022 Jul 11. Epub 2022 Jul 11.

Department of Materials Science and Engineering, National Tsing Hua University, Hsinchu 30013, Taiwan.

Recently, conductive-bridging memristors based on metal halides, such as halide perovskites, have been demonstrated as promising components for brain-inspired hardware-based neuromorphic computing. However, realizing devices that simultaneously fulfill all of the key merits (low operating voltage, high dynamic range, multilevel nonvolatile storage capability, and good endurance) remains a great challenge. Herein, we describe lead-free cesium halide memristors incorporating a MoO interfacial layer as a type of conductive-bridging memristor. With this design, we obtained highly uniform and reproducible memristors that exhibited all-around resistive switching characteristics: ultralow operating voltages (<0.18 V), low variations (<30 mV), long retention times (>10 s), high endurance (>10, full on/off cycles), record-high on/off ratios (>10, smaller devices having areas <5 × 10 mm), fast switching (<200 ns), and multilevel programming abilities (>64 states). With these memristors, we successfully implemented stateful logic functions in a reconfigurable architecture and accomplished a high classification accuracy ( 90%) in the simulated hand-written-digits classification task, suggesting their versatility in future in-memory computing applications. In addition, we exploited the room-temperature fabrication of the devices to construct a fully functional three-dimensional stack of memristors, which demonstrates their potential of high-density integration desired for data-intensive neuromorphic computing. High-performance, environmentally friendly cesium halide memristors provide opportunities toward next-generation electronics beyond von Neumann architectures.
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http://dx.doi.org/10.1021/acsnano.2c05436DOI Listing
July 2022

Epididymis cell atlas in a patient with a sex development disorder and a novel gene mutation.

Asian J Androl 2022 May 6. Epub 2022 May 6.

Institute of Reproductive Medicine, Medical School of Nantong University, Nantong 226000, China.

This study aims to characterize the cell atlas of the epididymis derived from a 46,XY disorders of sex development (DSD) patient with a novel heterozygous mutation of the nuclear receptor subfamily 5 group A member 1 (NR5A1) gene. Next-generation sequencing found a heterozygous c.124C>G mutation in NR5A1 that resulted in a p.Q42E missense mutation in the conserved DNA-binding domain of NR5A1. The patient demonstrated feminization of external genitalia and Tanner stage 1 breast development. The surgical procedure revealed a morphologically normal epididymis and vas deferens but a dysplastic testis. Microfluidic-based single-cell RNA sequencing (scRNA-seq) analysis found that the fibroblast cells were significantly increased (approximately 46.5%), whereas the number of main epididymal epithelial cells (approximately 9.2%), such as principal cells and basal cells, was dramatically decreased. Bioinformatics analysis of cell-cell communications and gene regulatory networks at the single-cell level inferred that epididymal epithelial cell loss and fibroblast occupation are associated with the epithelial-to-mesenchymal transition (EMT) process. The present study provides a cell atlas of the epididymis of a patient with 46,XY DSD and serves as an important resource for understanding the pathophysiology of DSD.
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http://dx.doi.org/10.4103/aja202226DOI Listing
May 2022

All-Vacuum-Deposited Perovskite X-ray Detector with a Record-High Self-Powered Sensitivity of 1.2 C Gy cm.

ACS Appl Mater Interfaces 2022 May 13;14(17):19795-19805. Epub 2022 Apr 13.

Department of Materials Science and Engineering, National Tsing Hua University, Hsinchu 30013, Taiwan.

Highly sensitive X-ray detection is crucial in, for example, medical imaging and secure inspection. Halide perovskite X-ray detectors are promising candidates for detecting highly energetic radiation. In this report, we describe vacuum-deposited Cs-based perovskite X-ray detectors possessing a p-i-n architecture. Because of the built-in potential of the p-i-n structure, these perovskite X-ray detectors were capable of efficient charge collection and displayed an exceptionally high X-ray sensitivity (1.2 C Gy cm) under self-powered, zero-bias conditions. We ascribe the outstanding X-ray sensitivity of the vacuum-deposited CsPbIBr devices to their prominent charge carrier mobility. Moreover, these devices functioned with a lowest detection limit of 25.69 nGy s and possessed excellent stability after exposure to over 3000 times the total dose of a chest X-ray image. For comparison, we also prepared traditional spin-coated CHNH-based perovskite devices having a similar device architecture. Their volume sensitivity was only one-fifth of that of the vacuum-deposited CsPbIBr devices. Thus, all-vacuum deposition appears to be a new strategy for developing perovskite X-ray detectors; with a high practical deposition rate, a balance can be reached between the thickness of the absorbing layer and the fabrication time.
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http://dx.doi.org/10.1021/acsami.2c03114DOI Listing
May 2022

Activation of PTH1R alleviates epididymitis and orchitis through Gq and β-arrestin-1 pathways.

Proc Natl Acad Sci U S A 2021 11;118(45)

Key Laboratory Experimental Teratology of the Ministry of Education and Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Cheeloo College of Medicine, Shandong University, Shandong 250012, China;

Inflammation in the epididymis and testis contributes significantly to male infertility. Alternative therapeutic avenues treating epididymitis and orchitis are expected since current therapies using antibiotics have limitations associated to side effects and are commonly ineffective for inflammation due to nonbacterial causes. Here, we demonstrated that type 1 parathyroid hormone receptor (PTH1R) and its endogenous agonists, parathyroid hormone (PTH) and PTH-related protein (PTHrP), were mainly expressed in the Leydig cells of testis as well as epididymal epithelial cells. Screening the secretin family G protein-coupled receptor identified that PTH1R in the epididymis and testis was down-regulated in mumps virus (MuV)- or lipopolysaccharide (LPS)-induced inflammation. Remarkably, activation of PTH1R by abaloparatide (ABL), a Food and Drug Administration-approved treatment for postmenopausal osteoporosis, alleviated MuV- or LPS-induced inflammatory responses in both testis and epididymis and significantly improved sperm functions in both mouse model and human samples. The anti-inflammatory effects of ABL were shown to be regulated mainly through the Gq and β-arrestin-1 pathway downstream of PTH1R as supported by the application of ABL in and mouse models. Taken together, our results identified an important immunoregulatory role for PTH1R signaling in the epididymis and testis. Targeting to PTH1R might have a therapeutic effect for the treatment of epididymitis and orchitis or other inflammatory disease in the male reproductive system.
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http://dx.doi.org/10.1073/pnas.2107363118DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8609314PMC
November 2021

Primary tumor immune score fails to predict the prognosis of colorectal cancer liver metastases after hepatectomy in Chinese populations.

Ann Transl Med 2021 Feb;9(4):310

State Key Laboratory of Oncology in South China, Sun Yat-sen University Cancer Center, Collaborative Innovation Center for Cancer Medicine, Guangzhou, China.

Background: Increasing evidence suggests that the immune score is significantly associated with cancer prognosis. However, the prognostic role of primary tumor immune score in colorectal cancer liver metastases (CRLM) after hepatectomy in Chinese patients has not been reported. The present study is designed to investigate whether the immune score of primary tumor can predict the postoperative survival of liver metastases in Chinese patients.

Methods: A total of 131 patients diagnosed with CRLM were included, and the corresponding primary tumor and liver metastasis specimens were acquired. An immune score ranging from 0 to 4 was established based on the counts and densities of CD3 and CD8 T cells in the core tumor (CT) and the invasive margin (IM). Relapse-free survival (RFS) and overall survival (OS) were analyzed by Kaplan-Meier curves to assess the prognostic role of primary tumor immune score. Furthermore, we conducted a comprehensive search of the Gene Expression Omnibus (GEO) and selected stage IV colorectal cancer (CRC) patients with liver metastasis to compare the tumor-infiltrating T cell profiles of the primary tumor and liver metastases by CIBERSORT.

Results: Patients with high immune scores in the primary tumor has no significantly better RFS and OS after hepatectomy than those with low immune scores [median RFS (95% CI): 19.13 (10.07-28.20) . 27.13 (15.97-38.29) months, P=0.604; median OS (95% CI): 64.37 (35.96-92.78) . 40.07 (32.54-47.59) months, P=0.652]. Data collected from the GEO indicates that the proportion of CD8 T cells and total T cells in the primary tumor and liver metastatic lesion are also not significantly correlated (CD8 T cells: r =0.030, P=0.468; total T cells: r =0.165, P=0.076).

Conclusions: The immune score of the primary tumor fails to predict the prognosis of CRLM after hepatectomy in Chinese patients.
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http://dx.doi.org/10.21037/atm-20-4932DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7944305PMC
February 2021

A novel prognostic nomogram for colorectal cancer liver metastasis patients with recurrence after hepatectomy.

Cancer Med 2021 03 4;10(5):1535-1544. Epub 2021 Feb 4.

State Key Laboratory of Oncology in South China, Sun Yat-sen University Cancer Center, Collaborative Innovation Center for Cancer Medicine, Guangzhou, People's Republic of China.

Purpose: We aimed to construct a nomogram to predict personalized post-recurrence survival (PRS) among colorectal cancer liver metastasis (CRLM) patients with post-hepatectomy recurrence.

Methods: Colorectal cancer liver metastasis patients who received initial hepatectomy and had subsequent recurrence between 2001 and 2019 in Sun Yat-sen University Cancer Center from China were included in the study. Patients were randomly assigned to a training cohort and a validation cohort on a ratio of 2:1. Univariable analysis was first employed to select potential predictive factors for PRS. Then, the multivariable Cox regression model was applied to recognize independent prognostic factors. According to the model, a nomogram to predict PRS was established. The nomogram's predictive capacity was further assessed utilizing concordance index (C-index) values, calibration plots, and Kaplan-Meier curves.

Results: About 376 patients were finally enrolled, with a 3-year PRS rate of 37.3% and a 5-year PRS rate of 24.6%. The following five independent predictors for PRS were determined to construct the nomogram: the largest size of liver metastases at initial hepatectomy, relapse-free survival, CEA level at recurrence, recurrent sites, and treatment for recurrence. The nomogram displayed fairly good discrimination and calibration. The C-index value was 0.742 for the training cohort and 0.773 for the validation cohort. Patients were grouped into three risk groups very well by the nomogram, with 5-year PRS rates of 45.2%, 23.3%, and 9.0%, respectively (p < 0.001) in the training cohort and 36.0%, 9.2%, and 4.6%, respectively (p < 0.001) in the validation cohort.

Conclusion: A novel nomogram was built and validated to enable the prediction of personal PRS in CRLM patients with post-hepatectomy recurrence. The nomogram may help physicians in decision making.
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http://dx.doi.org/10.1002/cam4.3697DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7940234PMC
March 2021

A Novel Ferroptosis-related Gene Signature for Overall Survival Prediction in Patients with Hepatocellular Carcinoma.

Int J Biol Sci 2020 6;16(13):2430-2441. Epub 2020 Jul 6.

State Key Laboratory of Oncology in South China, Sun Yat-sen University Cancer Center, Collaborative Innovation Center for Cancer Medicine, Guangzhou, People's Republic of China.

Hepatocellular carcinoma (HCC) is a highly heterogeneous disease, which makes the prognostic prediction challenging. Ferroptosis, an iron-dependent form of regulated cell death, can be induced by sorafenib. However, the prognostic value of ferroptosis-related genes in HCC remains to be further elucidated. In this study, the mRNA expression profiles and corresponding clinical data of HCC patients were downloaded from public databases. The least absolute shrinkage and selection operator (LASSO) Cox regression model was utilized to construct a multigene signature in the TCGA cohort. HCC patients from the ICGC cohort were used for validation. Our results showed that most of the ferroptosis-related genes (81.7%) were differentially expressed between HCC and adjacent normal tissues in the TCGA cohort. Twenty-six differentially expressed genes (DEGs) were correlated with overall survival (OS) in the univariate Cox regression analysis (all adjusted < 0.05). A 10-gene signature was constructed to stratify patients into two risk groups. Patients in the high-risk group showed significantly reduced OS compared with patients in the low-risk group ( < 0.001 in the TCGA cohort and = 0.001 in the ICGC cohort). The risk score was an independent predictor for OS in multivariate Cox regression analyses (HR> 1, < 0.01). Receiver operating characteristic (ROC) curve analysis confirmed the signature's predictive capacity. Functional analysis revealed that immune-related pathways were enriched, and immune status were different between two risk groups. In conclusion, a novel ferroptosis-related gene signature can be used for prognostic prediction in HCC. Targeting ferroptosis may be a therapeutic alternative for HCC.
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http://dx.doi.org/10.7150/ijbs.45050DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7378635PMC
November 2021

Clinical outcomes of microdissection testicular sperm extraction-intracytoplasmic sperm injection with fresh or cryopreserved sperm in patients with nonobstructive azoospermia.

Asian J Androl 2021 Mar-Apr;23(2):211-214

Department of Urology, Peking University Third Hospital, Beijing 100191, China.

We performed this study to evaluate the clinical outcomes of microdissection testicular sperm extraction-intracytoplasmic sperm injection (micro-TESE-ICSI) treatment that used fresh or cryopreserved sperm in patients with nonobstructive azoospermia (NOA). A total of 338 NOA patients with 344 consecutive cycles received treatment in the reproductive medicine center of Peking University Third Hospital in Beijing, China, from January 2014 to December 2017. Fresh oocytes and fresh sperm were used in 222 patients with 234 cycles (Group A). Fresh oocytes and cryopreserved sperm were used in 116 patients with 110 cycles (Group B). We compared patient characteristics, embryonic development, and pregnancy outcomes between Groups A and B. There was no statistical difference in the patient characteristics, and no differences were observed with fertilization or quality embryo rates between Groups A and B. The rates of clinical pregnancy and live birth were both higher for Group A than those for Group B (both P < 0.05). In conclusion, fresh testicular sperm appears to produce better ICSI outcomes than cryopreserved testicular sperm in patients with NOA.
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http://dx.doi.org/10.4103/aja.aja_38_20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7991819PMC
November 2021

Sperm retrieval rates and clinical outcomes for patients with different causes of azoospermia who undergo microdissection testicular sperm extraction-intracytoplasmic sperm injection.

Asian J Androl 2021 Jan-Feb;23(1):59-63

Department of Urology, Peking University Third Hospital, Beijing 100191, China.

The aim of our study was to compare the sperm retrieval rates (SRRs) and clinical outcomes of patients with different causes of azoospermia who underwent microdissection testicular sperm extraction-intracytoplasmic sperm injection (micro-TESE-ICSI). We conducted a retrospective study at the Reproductive Medicine Center of Peking University Third Hospital in Beijing, China, from January 2014 to December 2017. This study examined 769 patients with nonobstructive azoospermia who underwent 347 cycles of micro-TESE-ICSI. Patients with azoospermia were classified into Group A (Klinefelter syndrome, n = 284, 125 cycles), Group B (azoospermia Y chromosome factor c [AZFc] microdeletion, n = 91, 64 cycles), Group C (cryptorchidism, n = 52, 39 cycles), Group D (previous mumps and bilateral orchitis, n = 23, 23 cycles), and Group E (idiopathic azoospermia, n = 319, 96 cycles). Clinical characteristics, SRR, embryonic development, and pregnancy outcomes of the patients were compared between all groups. Patients in Group D had the highest and most successful SRR. The average SRR for all patients was 46.0%. The rates of clinical pregnancy, implantation, and live birth in Group D were 78.3%, 65.0%, and 74.0%, respectively, which were higher than those in all other groups (P < 0.05). Group B patients had the lowest clinical pregnancy, implantation, and live birth rates of all groups (P < 0.05). No differences were found in the miscarriage rate or birth defects among the groups (P > 0.05). Patients with orchitis had the highest SRR and best clinical outcomes. Although AZFc microdeletion patients had a higher SRR, their clinical outcomes were worse.
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http://dx.doi.org/10.4103/aja.aja_12_20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7831829PMC
October 2021

Altered microRNA profiles of testicular biopsies from patients with nonobstructive azoospermia.

Asian J Androl 2020 Jan-Feb;22(1):100-105

Department of Urology, Peking University Third Hospital, Beijing 100191, China.

Many studies have shown that microRNAs (miRNAs) play vital roles during the spermatogenesis. However, little is known about the altered miRNA profiles of testicular tissues in nonobstructive azoospermia (NOA). Using microarray technology, the miRNA expression profiles of testicular biopsies from patients with NOA and of normal testicular tissues were determined. Bioinformatics analyses were conducted to predict the enriched biological processes and functions of identified miRNAs. The microarray data were validated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), the results of which were then validated with a larger sample size. Correlations between the miRNA expression levels and clinical characteristics were analyzed. Receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic ability of miRNAs for azoospermia. Hierarchical clustering showed that 129 miRNAs were significantly differentially expressed between the NOA and control groups. Bioinformatics analysis indicated that the differentially expressed miRNAs were involved in spermatogenesis, cell cycle, and mitotic prometaphase. In the subsequent qRT-PCR assays, the selected miRNA expression levels were consistent with the microarray results, and similar validated results were obtained with a larger sample size. Some clinical characteristics were significantly associated with the expression of certain miRNAs. In particular, we identified a combination of two miRNAs (miR-10b-3p and miR-34b-5p) that could serve as a predictive biomarker of azoospermia. This study provides altered miRNA profiles of testicular biopsies from NOA patients and examines the roles of miRNAs in spermatogenesis. These profiles may be useful for predicting and diagnosing the presence of testicular sperm in individuals with azoospermia.
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http://dx.doi.org/10.4103/aja.aja_35_19DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6958976PMC
November 2020

Correlation between immune signature and high-density lipoprotein cholesterol level in stage II/III colorectal cancer.

Cancer Med 2019 03 7;8(3):1209-1217. Epub 2019 Feb 7.

Sate key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center, Guangzhou, P.R. China.

An increasing amount of evidence suggests that high-density lipoprotein cholesterol (HDL-C) is related to a positive prognosis in various cancers. However, the correlation between HDL-C and the immune signature and the prognostic role of HDL-C in stage II/III colorectal cancer (CRC) has not been previously reported. A total of 667 CRC patients were enrolled and divided into two groups based on the lower limit of normal HDL-C values (0.78 mmol/L). We used Kaplan-Meier curves and the Cox regression model to analyze the prognostic role of HDL in both disease-free survival (DFS) and overall survival (OS). Fifty-five pairs of tumor tissues were selected according to the variation in HDL-C levels (high or low) and the matched characterizes (ages, T stage, and N stage). Using immunohistochemistry, tumor tissues were stained with antibodies against CD3, CD8, CD163, iNOS, Forkhead box P3 (FOXP3), and CD33. We calculated the density of positively-stained infiltrating cells in the tumor center (TC) and invasive margin (IM). We then used Spearman rank correlation to further investigate the relationship between HDL-C levels and the immune signatures. Our results revealed that compared to patients with high HDL-C levels, patients with low HDL-C levels had poor 3-year DFS (68.9% vs 83.1%, P = 0.032) and 5-year OS rates (66.6% vs 85.3%, P = 0.002). We also identified a positive correlation between HDL-C and CD3 , CD8 and iNOS cells and a negative correlation between HDL-C and CD163 cells in both the TC and IM. This study reveals that a low HDL-C level in stage II/III CRC patients predicts poor prognosis. The correlation between the HDL-C level and immune signature in tissue specimens suggested that HDL-C is likely to play an inhibitory role in tumor development via affecting immune responses.
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http://dx.doi.org/10.1002/cam4.1987DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6434197PMC
March 2019

Ambient ozone pollution is associated with decreased semen quality: longitudinal analysis of 8945 semen samples from 2015 to 2018 and during pollution-control period in Beijing, China.

Asian J Androl 2019 Sep-Oct;21(5):501-507

Department of Urology, Peking University Third Hospital, Beijing 100191, China.

Previous studies suggest that air pollution has a negative effect on semen quality. However, most studies are cross-sectional and the results are controversial. This study investigated the associations between air pollutants (PM, PM, SO, NO, CO, and O) and semen quality among sperm donation candidates, especially when the air pollution was artificially controlled in Beijing, China. We analyzed 8945 semen samples in the human sperm bank of Peking University Third Hospital (Beijing, China) from October 2015 to May 2018. Air pollution data during the entire period (0-90 days prior) and key stages (0-9, 10-14, and 70-90 days prior) of sperm development were collected from the China National Environmental Monitoring Centre. The association between air pollutants and semen parameters (sperm concentration and progressive motility) was analyzed by a mixed model adjusted for age, abstinence duration, month, and average ambient temperature. Only Oduring key stages of 0-9 days and 10-14 days and the entire period was negatively associated with sperm concentration between 2015 and 2018 (P < 0.01). During the period of air pollution control from November 2017 to January 2018, except for the increase in Oconcentration, other five pollutants' concentrations decreased compared to those in previous years. In this period, the sperm concentration decreased (P < 0.001). During the pollution-control period, Oexposure 10-14 days prior was negatively associated with sperm concentration (95% CI: -0.399--0.111; P < 0.001). No significant association was found between the other five pollutants and semen quality during that period. Our study suggested that only Oexposure was harmful to semen quality. Therefore, Oshould not be neglected during pollution control operation.
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http://dx.doi.org/10.4103/aja.aja_116_18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6732886PMC
August 2020

Nanotechnology-assisted adipose-derived stem cell (ADSC) therapy for erectile dysfunction of cavernous nerve injury: cell tracking, optimized injection dosage, and functional evaluation.

Asian J Androl 2018 Sep-Oct;20(5):442-447

Department of Urology, Peking University Third Hospital, Beijing 100191, China.

Stem cell therapy is a potentially promising option for erectile dysfunction; however, its risk of tumorigenicity is a clinical hurdle and the risk is positively related to the number of injected cells. Our previous study showed that nanotechnology improved adipose-derived stem cell (ADSC) therapy for erectile dysfunction of cavernous nerve injury (CNI) by attracting cells in the corpus cavernosum. These results indicated the possibility of using a reduced dosage of ADSCs for intracavernous injection. In this exploratory study, we used lower dosage (2 × 10 cells) of ADSCs for intracavernous injection (ICI) and the nanotechnology approach. Intracavernous pressure and mean arterial pressure were measured at day 28 to assess erectile function. The low-dose ADSC therapy group showed favorable treatment effects, and nanotechnology further improved these effects. In vivo imaging of ICI cells revealed that the fluorescein signals of NanoShuttle-bound ADSCs (NanoADSCs) were much stronger than those of ADSCs at days 0, 1, and 3. Both immunofluorescence and Western blot analysis showed a significant increase in smooth muscle, endothelium, and nerve tissue in the ADSC group compared to that in the CNI group; further improvement was achieved with assisted nanotechnology. These findings demonstrate that nanotechnology can be used to further improve the effect of small dosage of ADSCs to improve erectile function. Abundant NanoADSCs remain in the corpus cavernosum in vivo for at least 3 days. The mechanism of erectile function improvement may be related to the regeneration of the smooth muscle, endothelium, and nerve tissues.
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http://dx.doi.org/10.4103/aja.aja_48_18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6116694PMC
September 2019

Evaluation of the long-term safety and effectiveness of tadalafil once daily in Chinese men with erectile dysfunction: interim results of a multicenter, randomized, open-label trial.

Asian J Androl 2018 Nov-Dec;20(6):587-592

Peking University First Hospital, Institute of Urology, Peking University, Beijing 100034, China.

Once-daily tadalafil administration has been well established; however, studies about tadalafil once-daily treatment in the Chinese population are lacking. In this phase 4, postmarketing study, we ascertained the long-term safety and effectiveness of tadalafil 2.5 mg and 5.0 mg once daily in Chinese men with erectile dysfunction (n = 635). The primary endpoint of the study was safety at 12 months as assessed by the proportion of patients experiencing at least one treatment-emergent adverse event (serious or nonserious). The secondary endpoints included safety and effectiveness, measured by the International Index of Erectile Function-Erectile Function (IIEF-EF) domain scores. Similar adverse events to the known safety profile of tadalafil, such as nasopharyngitis, upper respiratory tract infection, headache, and dizziness, were detected. No new cardiovascular safety concerns were observed. After 3 months of treatment, significant increases in IIEF-EF domain scores were detected for both 2.5-mg (least squares [LS] mean change: 6.3; 95% confidence interval [CI]: 5.4-7.1; P < 0.001) and 5.0-mg (LS mean change: 7.4; 95% CI: 6.8-7.9; P < 0.001) tadalafil doses, and significance was maintained up to 12 months. In addition, approximately 40% of patients regained normal erectile function (IIEF-EF ≥26) following 1 year of tadalafil once-daily treatment. The findings in this study provide evidence for the extended effectiveness and tolerability of tadalafil, demonstrating no new safety concerns, in a Chinese population and make once-daily tadalafil administration a viable option for improving sexual performance and satisfaction in Chinese men with erectile dysfunction.
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http://dx.doi.org/10.4103/aja.aja_47_18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6219303PMC
October 2019

A clinical trial on the consistency of bilateral testicular tissue histopathology and Johnsen score: single side or bilateral side biopsy?

Oncotarget 2018 May 8;9(35):23848-23859. Epub 2018 May 8.

Department of Urology, Peking University Third Hospital, Beijing 100191, China.

To evaluate and compare left and right testicular tissue histopathology and Johnsen score, and to investigate the necessity for bilateral testicular biopsy. We recruited180 patients with non-obstructiveazoospermia (NOA) on testicular biopsy who had undergonetesticular sperm aspiration (TESA). Pathological sections of testicular tissue were diagnosed by specially-assigned doctors, who evaluated pathological findings, determined the Johnsen score and confirmed for the presence or absence of sperm. Sperm positive rates for left and right testicular histopathology were 55.0% and 51.7% respectively, and the proportion of Johnsen scores≥8 for left and right testes were 53.3% and 50.0%, respectively. Cohen kappa values revealed that the identification of sperm in bilateral testicular samples was not consistent and was related to random effects; Optimized cut-off value for bilateral testicular volume was 11ml (Johnsen score ≥8), and optimized cut-off values of E on left and right testes were 144.5pmol/L and 133.5 pmol/L (Johnsen score≤7). However, age, serum prolactin (PRL), follicle stimulating hormone (FSH), luteinizing hormone (LH) and total testosterone (TT) levels were not accurate predictors for the existence of testicular sperm. There was nostatistical significance between left and right testicular histopathology in terms of sperm positive rates or Johnsen score; the Johnsen score were caused entirely by random effects and a score from one side could not represent the other side. Therefore, we recommend that both testes need to undergo surgery when NOA patients undergo testicular biopsy or sperm retrieval.
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http://dx.doi.org/10.18632/oncotarget.24748DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5963628PMC
May 2018

[Microdissection testicular sperm extraction for patients with nonmosaic Klinefelter syndrome: A report of 143 cases].

Zhonghua Nan Ke Xue 2017 Dec;23(12):1075-1079

Center of Reproductive Medicine, Peking University Third Hospital, Beijing 100191, China.

Objective: To investigate the clinical application and outcomes of microdissection testicular sperm extraction (micro-TESE) in patients with nonmosaic Klinefelter syndrome (KS).

Methods: A total of 143 nonmosaic KS patients underwent micro-TESE in the Center of Reproductive Medicine of Peking University Third Hospital between July 2012 and August 2016. We analyzed their clinical and follow-up data and evaluated the outcomes.

Results: Spermatozoa were successfully retrieved from the testicular tissue in 44.76% (64/143) of the patients, 84.4% (54/64) by unilateral and 15.6% (10/64) by bilateral micro-TESE. Seventy-five of the KS patients were followed up in the years of 2014 and 2015. Of the 34 patients with successful sperm retrieval, 73.52% (25/34) achieved clinical pregnancy and 8 boys and 8 girls were already born in 14 of the 25 cases.

Conclusions: The micro-TESE is a useful method for sperm retrieval in nonmosaic KS patients, with high rates of sperm retrieval, clinical pregnancy, and birth of biological offspring.
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December 2017

[Collagenase clostridium histolyticum for Peyronie's disease: A new minimally invasive and effective treatment].

Zhonghua Nan Ke Xue 2017 Sep;23(9):771-775

Department of Urology, Peking University Third Hospital, Beijing 100191, China .

The treatment of Peyronie's disease has been a challenge to urologists, as most of the current oral medications are considered hardly valid and the rest of the therapies require strict control of indications, some of which remain controversial, while the final surgical option may be associated with complications such as penile shortening or erectile dysfunction. Collagenase clostridium histolyticum has been proved to be effective in dissolving the penile cavernosal plaque and approved by the Food and Drug Administration of the USA as the first drug to be used for the treatment of Peyronie's disease. This article presents an introduction to the characteristics, safety, efficacy, and procedures of this new treatment, which may benefit urologists and the patients with Peyronie's disease.
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September 2017

Ran-binding protein M is associated with human spermatogenesis and oogenesis.

Mol Med Rep 2018 Feb 23;17(2):2257-2262. Epub 2017 Nov 23.

1Department of Urology, The Third Hospital of Peking University, Beijing 100191, P.R. China.

The aim of the present study was to explore the underlying mechanism and diagnostic potential of Ran‑binding protein M (RanBPM) in human spermatogenesis and oogenesis. RanBPM expression in human testis and ovaries was analysed using polymerase chain reaction (PCR) and western blotting, and immunofluorescence was performed on testis and ovary tissue sections during different developmental stages of spermatogenesis and oogenesis using RanBPM antibodies. Interactions with a variety of functional proteins were also investigated. RanBPM mRNA and protein expression levels were determined by PCR and western blotting in the tissue sections. Results revealed that the mRNA expression levels were highest in the testis followed by the ovary. The RanBPM protein was predominantly localized in the nucleus of germ cells, and the expression levels were highest in pachytene spermatocytes and cells surrounding spermatids in testis tissue. In ovary cells, RanBPM was localized in the nucleus and cytoplasm. In conclusion, the results suggested that RanBPM may have multiple roles in the regulation of germ cell proliferation during human spermatogenesis and oogenesis. This research may provide a novel insight into the underlying molecular mechanism of RanBPM and may have implications for the clinical diagnosis and treatment of human infertility.
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http://dx.doi.org/10.3892/mmr.2017.8147DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5783472PMC
February 2018

The Immunoscore system predicts prognosis after liver metastasectomy in colorectal cancer liver metastases.

Cancer Immunol Immunother 2018 03 4;67(3):435-444. Epub 2017 Dec 4.

State Key Laboratory of Oncology in South China, Sun Yat-sen University Cancer Center, Collaborative Innovation Center for Cancer Medicine, 651# Dongfeng Road East, Guangzhou, 510060, People's Republic of China.

Background: The Immunoscore was initially established to evaluate the prognosis of stage I/II/III colorectal cancer patients. However, the feasibility of the Immunoscore for the prognosis of colorectal cancer liver metastases (CRCLM) has not been reported.

Methods: Liver metastases in 249 CRCLM patients were retrospectively analyzed. The Immunoscore was assessed according to the counts and densities of CD3+ and CD8+ T cells in the central- and peritumoral areas by immunohistochemistry. The prognostic role of the Immunoscore for relapse-free survival (RFS) and overall survival (OS) was analyzed with Kaplan-Meier curves and Cox multivariate models, and confirmed via an internal validation. Receiver operating characteristic (ROC) curves were plotted to compare the prognostic values of the Immunoscore and the clinical risk score (CRS) system.

Results: CRCLM patients with high Immunoscores (> 2) had significantly longer RFS [median RFS (95% confidence interval; 95% CI) 21.4 (7.8-35.1) vs. 8.7 (6.8-10.5) months, P < 0.001] and OS [median OS (95% CI): not reached vs. 28.7 (23.2-34.2) months, P < 0.001] than those with low Immunoscores (≤ 2). After stratification by CRS, the Immunoscore retained a statistically significant prognostic value for OS. The areas under the ROC curves (AUROCs) of the Immunoscore and the CRS system for RFS were 0.711 [95% CI 0.642-0.781] and 0.675[95% CI 0.601-0.749] (P = 0.492), whereas the AUROC of the Immunoscore system for OS was larger than that of the CRS system [0.759 (95% CI 0.699-0.818) vs. 0.660 (95% CI 0.592-0.727); P = 0.029].

Conclusions: The Immunoscore of liver metastases can be applied to predict the prognosis of CRCLM patients following liver resection.
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http://dx.doi.org/10.1007/s00262-017-2094-8DOI Listing
March 2018

Pathologic response after preoperative therapy predicts prognosis of Chinese colorectal cancer patients with liver metastases.

Chin J Cancer 2017 Oct 2;36(1):78. Epub 2017 Oct 2.

Sun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou, 510060, Guangdong, P. R. China.

Background: Pathologic response is evaluated according to the extent of tumor regression and is used to estimate the efficacy of preoperative treatment. Several studies have reported the association between the pathologic response and clinical outcomes of colorectal cancer patients with liver metastases who underwent hepatectomy. However, to date, no data from Chinese patients have been reported. In this study, we aimed to evaluate the association between the pathologic response to pre-hepatectomy chemotherapy and prognosis in a cohort of Chinese patients.

Patients And Methods: In this retrospective study, we analyzed the data of 380 liver metastases in 159 patients. The pathologic response was evaluated according to the tumor regression grade (TRG). The prognostic role of pathologic response in recurrence-free survival (RFS) and overall survival (OS) was assessed using Kaplan-Meier curves with the log-rank test and multivariate Cox models. Factors that had potential influence on pathologic response were also analyzed using multivariate logistic regression and Kruskal-Wallis/Mann-Whitney U tests.

Results: Patients whose tumors achieved pathologic response after preoperative chemotherapy had significant longer RFS and OS than patients whose tumor had no pathologic response to chemotherapy (median RFS: 9.9 vs. 6.5 months, P = 0.009; median OS: 40.7 vs. 28.1 months, P = 0.040). Multivariate logistic regression and Kruskal-Wallis/Mann-Whitney U tests showed that metastases with small diameter, metastases from the left-side primary tumors, and metastases from patients receiving long-duration chemotherapy had higher pathologic response rates than their control metastases (all P < 0.05). A decrease in the serum carcinoembryonic antigen (CEA) level after preoperative chemotherapy predicted an increased pathologic response rate (P < 0.05). Although the application of targeted therapy did not significantly influence TRG scores of all cases of metastases, the addition of cetuximab to chemotherapy resulted in a higher pathologic response rate when combined with irinotecan-based regimens rather than with oxaliplatin-based regimens.

Conclusions: We found that the evaluation of pathologic response may predict the prognosis of Chinese colorectal cancer patients with liver metastases after preoperative chemotherapy. Small tumor diameter, long-duration chemotherapy, left primary tumor, and decreased serum CEA level after chemotherapy are associated with increased pathologic response rates.
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http://dx.doi.org/10.1186/s40880-017-0244-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5625647PMC
October 2017

Penile rehabilitation with a vacuum erectile device in an animal model is related to an antihypoxic mechanism: blood gas evidence.

Asian J Androl 2013 May 8;15(3):387-90. Epub 2013 Apr 8.

Department of Urology, Affiliated Drum Tower Hospital, School of Medicine, Nanjing University, Nanjing 210008, China.

Our previous study showed that vacuum erectile device (VED) therapy has improved erectile function in rats with bilateral cavernous nerve crush (BCNC) injuries. This study was designed to explore the mechanism of VED in penile rehabilitation by analyzing cavernous oxygen saturation (SO2) and to examine the effect of VED therapy on preventing penile shrinkage after BCNC. Thirty adult Sprague-Dawley rats were randomly assigned into three groups: group 1, sham surgery; group 2, BCNC; and group 3, BCNC+VED. Penile length and diameter were measured on a weekly basis. After 4 weeks of therapy, the penile blood was extracted by three methods for blood gas analysis (BGA): method 1, cavernous blood was aspirated at the flaccid state; method 2, cavernous blood was aspirated at the traction state; and method 3, cavernous blood was aspirated immediately after applying VED. SO2 values were tested by the blood gas analyzer. The results showed that VED therapy is effective in preventing penile shrinkage induced by BCNC (Penile shortening: BCNC group 1.9±1.1 mm; VED group 0.3±1.0 mm; P<0.01. Penile diameter reduction: BCNC group 0.28±0.14 mm; VED group 0.04±0.14 mm; P<0.01). The mean SO2±s.d. values were increased by VED application (88.25%±4.94%) compared to the flaccid (76.53%±4.16%) or traction groups (78.93%±2.56%) (P<0.05). The calculated blood constructs in the corpus cavernosum right after VED application were 62% arterial and 38% venous blood. These findings suggest that VED therapy can effectively preserve penile size in rats with BCNC injury. The beneficial effect of VED therapy is related to antihypoxia by increasing cavernous blood SO2.
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http://dx.doi.org/10.1038/aja.2013.18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3739658PMC
May 2013

The effects of long-term administration of tadalafil on STZ-induced diabetic rats with erectile dysfunction via a local antioxidative mechanism.

Asian J Androl 2012 Jul 16;14(4):616-20. Epub 2012 Apr 16.

Department of Urology, the Affiliated Drum Tower Hospital, Nanjing University Medical School, Nanjing 210008, China.

Type 5 phosphodiesterase inhibitors (PDE5Is) are well known being effective via the nitric oxide and cyclic guanosine monophosphate (NO-cGMP) pathway and are widely used in the treatment of diabetic erectile dysfunction (ED). However, it is unclear whether other pathways may be involved in the treatment of diabetic ED with PDE5Is. The purpose of this study was to clarify the role of antioxidants in diabetic ED treatment through the long-term administration of PDE5Is. Three groups of Sprague-Dawley rats were utilized: Group N, the normal control; Group D, streptozotocin (STZ)-induced diabetic rats as a control; and Group D+T, STZ-induced diabetic rats who received oral administration of tadalafil for 8 weeks. Erectile function was assessed by intracavernous pressure (ICP) and mean arterial pressure (MAP) during electrical stimulation of the cavernous nerve before euthanasia. The levels of malondialdehyde (MDA), superoxide dismutase (SOD) and mitochondrial membrane potential (MMP) of cavernous tissue were assessed by biochemical analysis. The morphology of mitochondria was observed by electron microscopy. The ICP/MAP ratio was higher in Group D+T than in Group D (P<0.05). The levels of MDA decreased and the activities of SOD increased in Group D+T in comparison with Group D (P<0.05). The mitochondrial membrane potential level of cavernous tissue in diabetic rats was partially recovered by tadalafil treatment for 8 weeks. The morphology changes of mitochondria were also remarkably ameliorated in Group D+T. Collectively, the long-term administration of tadalafil in diabetic rats partially reduced oxidative stress lesions of the penis via a local antioxidative stress pathway. Long-term dosages of tadalafil given once daily beginning soon after the onset of diabetes may aid in preventing rats from developing diabetic ED.
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http://dx.doi.org/10.1038/aja.2012.22DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3720091PMC
July 2012

Mobilisation of endothelial progenitor cells: one of the possible mechanisms involved in the chronic administration of melatonin preventing erectile dysfunction in diabetic rats.

Asian J Androl 2012 May 27;14(3):481-6. Epub 2012 Feb 27.

Department of Urology, Affiliated Drum Tower Hospital, School of Medicine, Nanjing University, Nanjing, China.

Diabetes-induced oxidative stress plays a critical role in the mobilisation of endothelial progenitor cells (EPCs) from the bone marrow to the circulation. This study was designed to explore the effects of chronic melatonin administration on the promotion of the mobilisation of EPCs and on the preservation of erectile function in type I diabetic rats. Melatonin was administered to streptozotocin-induced type I diabetic rats. EPCs levels were determined using flow cytometry. Oxidative stress in the bone marrow was indicated by the levels of superoxide dismutase and malondialdehyde. Erectile function was evaluated by measuring the intracavernous pressure during an electrostimulation of the cavernous nerve. The density of the endothelium and the proportions of smooth muscle and collagen in the corpus cavernosum were determined by immunohistochemistry. The administration of melatonin increased the superoxide dismutase level and decreased the malondialdehyde level in the bone marrow. This effect was accompanied by an increased level of circulating EPCs in the diabetic rats. The intracavernous pressure to mean arterial pressure ratio of the rats in the treatment group was significantly greater, compared with diabetic control rats. The histological analysis demonstrated an increase in the endothelial density of the corpus cavernosum after the administration of melatonin. However, melatonin treatment did not change the proportions of smooth muscle and collagen in the corpus cavernosum of diabetic rats. Chronic administration of melatonin has a beneficial effect on preventing erectile dysfunction (ED) in type I diabetic rats. Promoting the mobilisation of EPCs is one of the possible mechanisms involved in the improvement of ED.
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http://dx.doi.org/10.1038/aja.2011.161DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3720169PMC
May 2012

Should patients with erectile dysfunction be evaluated for cardiovascular disease?

Asian J Androl 2012 Jan 28;14(1):138-44. Epub 2011 Nov 28.

Division of Urology, University of Texas Medical School at Houston, 77030, USA.

The landmark Massachusetts Male Ageing Study shed new light on the prevalence of erectile dysfunction (ED) and drew attention to ED as a disease of ageing. Over the years, ED has been linked to the development of cardiovascular disease (CVD) in some patients. There is clear evidence that ED and CVD share and have a similar risk factor profile. CVD is one of the most recognizable causes of mortality and early detection coupled with prevention of mortality from CVD has been the prime interest of many researchers. Consequently, there has been a multidisciplinary curiosity regarding the proposal to use ED as a marker for future CVD. In fact, there have been several proposals to use ED as a screening tool for future CVD. We performed a comprehensive search of two main databases-PubMed and Cochrane Library using a combination of keywords such as acute myocardial infarction, coronary artery disease (CAD) and ED. Journal articles from January 2000 to June 2011 were reviewed. We included all articles discussing the relationship between ED and CVD in the English language. All the relevant randomized controlled trials, cohort and retrospective studies, and review articles were included in our overall analysis in an attempt to answer the question whether all patients with ED should be clinically evaluated for CVD. The results showed a link between ED and the development of future CVD in some patients, but ED was not shown to be an independent risk predictor that is any better than the traditional Framingham risk factors. Screening for CVD may, however, be rewarding in younger patients with severe ED and in patients with concurrent CVD risk factors.
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http://dx.doi.org/10.1038/aja.2011.137DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3735161PMC
January 2012

Combination therapy for erectile dysfunction: an update review.

Asian J Androl 2011 May 21;13(3):382-90. Epub 2011 Mar 21.

Division of Urology, University of Texas Medical School at Houston, Houston, TX 77030, USA.

The introduction of oral phosphodiesterase-5 inhibitors (PDE5Is) in the late 1990s and early 2000s revolutionized the field of sexual medicine and PDE5Is are currently first-line monotherapy for erectile dysfunction (ED). However, a significant proportion of patients with complex ED will be therapeutic non-responders to PDE5I monotherapy. Combination therapy has recently been adopted for more refractory cases of ED, but a critical evaluation of current combination therapies is lacking. A thorough PubMed and Cochrane Library search was conducted focusing on the effectiveness of combination therapies for ED in therapeutic non-responders to PDE5I therapy. Journal articles spanning the time period between January 1990 and December 2010 were reviewed. Criteria included all pertinent review articles, randomized controlled trials, cohort studies and retrospective analyses. References from retrieved articles were also manually scanned for additional relevant publications. Published combination therapies include PDE5I plus vacuum erectile device (VED), intraurethral medication, intracavernosal injection (ICI), androgen supplement, α-blocker or miscellaneous combinations. Based on this review, some of these combination treatments appeared to be quite effective in preliminary testing. Caution must be advised, however, as the majority of combination therapy articles in the last decade have numerous limitations including study biases and small subject size. Regardless of limitations, present combination therapy research provides a solid foundation for future studies in complex ED management.
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http://dx.doi.org/10.1038/aja.2011.2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3739353PMC
May 2011

The impact of minimally invasive surgeries for the treatment of symptomatic benign prostatic hyperplasia on male sexual function: a systematic review.

Asian J Androl 2010 Jul 17;12(4):500-8. Epub 2010 May 17.

Division of Urology, University of Texas Medical School at Houston, Houston, TX 77030, USA.

A systematic review of randomized controlled trials and cohort studies was conducted to evaluate data for the effects of minimally invasive procedures for treatment of symptomatic benign prostatic hyperplasia (BPH) on male sexual function. The studies searched were trials that enrolled men with symptomatic BPH who were treated with laser surgeries, transurethral microwave therapy (TUMT), transurethral needle ablation of the prostate (TUNA), transurethral ethanol ablation of the prostate (TEAP) and high-intensity frequency ultrasound (HIFU), in comparison with traditional transurethral resection of the prostate (TURP) or sham operations. A total of 72 studies were identified, of which 33 met the inclusion criteria. Of the 33 studies, 21 were concerned with laser surgeries, six with TUMT, four with TUNA and two with TEAP containing information regarding male sexual function. No study is available regarding the effect of HIFU for BPH on male sexual function. Our analysis shows that minimally invasive surgeries for BPH have comparable effects to those of TURP on male erectile function. Collectively, less than 15.4% or 15.2% of patients will have either decrease or increase, respectively, of erectile function after laser procedures, TUMT and TUNA. As observed with TURP, a high incidence of ejaculatory dysfunction (EjD) is common after treatment of BPH with holmium, potassium-titanyl-phosphate and thulium laser therapies (> 33.6%). TUMT, TUNA and neodymium:yttrium aluminum garnet visual laser ablation or interstitial laser coagulation for BPH has less incidence of EjD, but these procedures are considered less effective for BPH treatment when compared with TURP.
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http://dx.doi.org/10.1038/aja.2010.33DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3739367PMC
July 2010

Prospective and long-term evaluation of erect penile length obtained with inflatable penile prosthesis to that induced by intracavernosal injection.

Asian J Androl 2009 Jul 15;11(4):411-5. Epub 2009 Jun 15.

Division of Urology, University of Texas Medical School at Houston, TX 77030, USA.

We compared erect penile length after inflatable penile prosthesis (IPP) implantation with that induced by in-tracavernosal injection (ICI) before surgery. A total of 11 patients with full erections induced by ICI of 0.25 mL of Trimix (papaverine, phentolamine and prostaglandin E1) at the time of penile color duplex Doppler ultrasonography were enrolled in this study. The patients subsequently underwent IPP implantation as the ultimate treatment for their erectile dysfunction (ED). Erect penile length consequent to IPP implantation was measured at 6 weeks, 6 months and at 1 year after surgery. The Sexual Health Inventory for Men (SHIM) was administered before, and at 6 months and 1 year after IPP implantation. Erect penile length (mean +/- s.e.) as induced by ICI was 13.2 +/- 0.4 cm, whereas the lengths attained with IPP were 12.4 +/- 0.3, 12.5 +/- 0.3 and 12.5 +/- 0.4 cm at the sixth week, sixth month and 1-year follow-ups, respectively. There were 0.83 +/- 0.25, 0.75 +/- 0.20 and 0.74 +/- 0.15 cm decreases in erect penile length at 6 weeks, 6 months and 1 year, respectively, after IPP implantation when compared with that after ICI (P < 0.05). The SHIM scores for patients reporting shorter penises were the same as those for patients without complaints at the 6-month and 1-year follow-ups (P > 0.05). To our knowledge, this is the first study to objectively show a significant decrease in erect penile length after IPP implantation when compared with that after ICI. However, this decrease did not affect the effectiveness of IPP in treating ED.
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http://dx.doi.org/10.1038/aja.2009.35DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3735309PMC
July 2009
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