A fully validated LC­MS/MS method for quantifying bevacizumab in plasma samples from patients with NSCLC and its implications in therapeutic drug monitoring.

Given the increasing use of bevacizumab in combinatorial drug therapy for a multitude of different cancer types, there is a need for therapeutic drug monitoring to analyze the possible correlation between drug trough concentration, and therapeutic effect and adverse reactions. An ultra-performance liquid chromatography tandem-mass spectrometry method was then developed and validated to determine bevacizumab levels in human plasma samples. Chromatographic separation was achieved on a Shimadzu InertSustainBio C18 HP column, whereas subsequent mass spectrometric analysis was performed using a Shimadzu 8050CL triple quadrupole mass spectrometer equipped with an electro-spray ionization source in the positive ion mode. In total, three multiple reaction monitoring transitions of each of the surrogate peptides were chosen with 'FTFSLDTSK' applied as the quantification peptide whereas 'VLIYFTSSLHSGVPSR' and 'STAYLQMNSLR' were designated as the verification peptides using the Skyline software. This analytical method was then fully validated, with specificity, linearity, lower limit of quantitation, accuracy, precision, stability, matrix effect and recovery calculated. The linearity of this method was developed to be within the concentration range 5-400 µg/ml for bevacizumab in human plasma. Subsequently, eight patients with non-small cell lung cancer (NSCLC) were recruited and injected with bevacizumab over three periods of treatment to analyze their steady-state trough concentration and differences. To conclude, the results of the present study suggest that bevacizumab can be monitored in a therapeutic setting in patients with NSCLC.

A fully validated method for simultaneous determination of icotinib, osimertinib, gefitinib and O-desmethyl gefitinib in human plasma using UPLC-MS/MS for therapeutic drug monitoring.

BACKGROUND AND AIMS: A few researches have reported the exposure-efficacy/toxicity relationships of epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs). On account of the large interpatient pharmacokinetic variability, therapeutic drug monitoring (TDM) seems promising for optimizing dosage regimen and improving treatment efficacy and safety. Therefore, a rapid and convenient ultrahigh performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the determination of icotinib, osimertinib, gefitinib and O-demesthyl gefitinib in human plasma for TDM. MATERIALS AND METHODS: Icotinib-D4 and osimertinib-13CD3 were used as the internal standards (ISs). The samples were prepared by protein precipitation using acetonitrile. Chromatographic separation was achieved on a 40 â„ƒ Shimadzu Shim-pack Scepter C18-120 column (2.1 ×50 mm, 3.0 µm, Japan) by a Shimadzu 30 A solvent management system. Detection was carried out using a Shimadzu LC-MS 8050CL triple quadrupole mass spectrometer coupled with an electrospray ionization source in positive mode. RESULTS: This analytical method was fully validated with selectivity, carry-over, linearity, lower limit of quantification, accuracy (from 92.68% to 106.62%) and precision (intra- and inter-day coefficients of variation ranged from 0.92% to 9.85%), matrix effect, extraction recovery, stability and dilution integrity. The calibration curves were developed to be within the concentration ranges of 200-4000 ng/mL for icotinib, 50-1000 ng/mL for osimertinib, gefitinib and O-desmethyl gefitinib in human plasma which meet the needs of routine TDM. CONCLUSIONS: The proposed method was used in 100 patients with non-small cell lung cancer for monitoring plasma concentration of the mentioned EGFR-TKIs. The trough concentrations of ICO were distributed between 226.42 ng/mL and 3853.36 ng/mL, peak concentrations were between 609.20 ng/mL and 2191.54 ng/mL. The trough concentrations of OSI were distributed between 110.48 ng/mL and 1183.13 ng/mL. The trough concentrations of GEF were distributed between 117.71 ng/mL and 582.74 ng/mL, while DeGEF was distributed from 76.21 ng/mL to 1939.83 ng/mL with two less than 20 ng/mL. The results of therapeutic drug monitoring aimed to investigate exposure-efficacy/toxicity relationship and improve the efficacy and safety of targeted therapies.

Severe autoimmune hemolytic anemia complicating hereditary spherocytosis treated successfully with glucocorticoids and cyclosporine: a case report.

BACKGROUND: Severe autoimmune hemolytic anemia complicating hereditary spherocytosis is life threatening and has not been described in a case report. Here, we report a case in which this intractable disease was treated successfully with glucocorticoids and cyclosporine. CASE PRESENTATION: A 25-year-old female patient with hereditary spherocytosis developed severe autoimmune hemolytic anemia after respiratory syncytial virus infection. Her hemoglobin level was 26 g/L and various anti-red blood cell antibodies were detected in her serum, making blood matching difficult. Glucocorticoid monotherapy was ineffective. With the addition of cyclosporine (50 mg/12 h), the patient's hemoglobin level increased significantly and the symptoms associated with anemia were greatly relieved. CONCLUSION: In patients with severe autoimmune hemolytic anemia, especially when the presence of multiple anti-red blood cell antibodies and alloantibodies interferes with blood matching, a glucocorticoid-cyclosporine regimen may be tried.

Case report: First case of pemetrexed plus cisplatin-induced immune hemolytic anemia in a patient with lung adenocarcinoma.

Background: Drug-induced immune hemolytic anemia (DIIHA) is a rare but potentially life-threatening drug-related complication. There are no previous reports of pemetrexed plus cisplatin as first-line chemotherapy for non-small cell lung cancer, resulting in DIIHA. Case presentation: In this report, a patient with advanced-stage lung adenocarcinoma developed severe immune hemolytic anemia 21 days after pemetrexed plus cisplatin chemotherapy. Laboratory findings showed severe hemolysis, including a rapid decrease in hemoglobin (HGB) and an elevated level of reticulocytes (Rets), indirect bilirubin (IBIL), and lactate dehydrogenase (LDH). A workup for the possibility of DIIHA was performed, including a direct antiglobulin test (DAT), a test in the presence of the soluble drug, and a drug-treated red blood cell (RBC) test. It showed a strongly positive (3+) result for anti-C3d but not for anti-immunoglobin G (IgG) in DAT. Enzyme-treated RBCs reacted weakly with the patient's serum and pemetrexed when complement was added. In addition, the patient's serum and normal sera were reactive with cisplatin-treated RBCs. However, eluates from the patient's RBCs and diluted normal sera were non-reactive with cisplatin-coated RBCs. Untreated and enzyme-treated RBCs reacted with the patient's serum in the presence of soluble cisplatin. In vitro serological tests suggested that complement-dependent pemetrexed antibodies and cisplatin-associated non-immunologic protein adsorption (NIPA) might combine to cause immune hemolytic anemia. The patient's anemia gradually recovered when pemetrexed and cisplatin were discontinued. Conclusion: This rare case demonstrated that complement-dependent pemetrexed antibodies and cisplatin-associated NIPA might occur simultaneously in a patient with DIIHA.

Effect of Dialysis on Antiplatelet Drug Efficacy in Uremic Patients with Coronary Heart Disease.

BACKGROUND: Coronary intervention therapy is the main treatment for uremic patients with coronary heart disease. The studies on whether dialysis reduces the efficacy of dual antiplatelet drugs are limited. The aim of this study was to examine the effect of dialysis on antiplatelet drugs in uremic patients with coronary heart disease. METHODS: This study included 26 uremic patients who had undergone percutaneous coronary intervention in China-Japan Friendship Hospital from November 2015 to May 2017. We examined their thromboelastography results before and after hemodialysis. Self-paired t-tests were employed to analyze changes in the inhibition rate of platelet aggregation. RESULTS: The mean inhibition rates of arachidonic acid-induced platelet aggregation before and after hemodialysis were 82.56 ± 2.79% and 86.42 ± 3.32%, respectively (t= -1.278, P= 0.213). The mean inhibition rates of adenosine diphosphate-induced platelet aggregation before and after hemodialysis were 67.87 ± 5.10% and 61.94 ± 5.90%, respectively (t = 1.425, P= 0.167). There was no significant difference in the inhibition rates of platelet aggregation before or after hemodialysis. These results also applied to patients with different sensitivity to aspirin and clopidogrel. CONCLUSION: Dialysis did not affect the antiplatelet effects of aspirin and clopidogrel in uremic patients with coronary heart disease.

Diagnostic value of interferon-γ-induced protein of 10kDa for tuberculous pleurisy: A meta-analysis.

BACKGROUND: The accurate diagnosis of tuberculous pleurisy is still a clinical challenge. Many studies reported that interferon-γ-induced protein of 10kDa (IP-10) plays a role in diagnosing tuberculous pleurisy, but with considerable variance of results. This meta-analysis aimed to evaluate the overall diagnostic accuracy of IP-10 for tuberculous pleurisy. METHODS: PubMed, EMBASE, and other databases were searched for studies examining accuracy of pleural IP-10 for diagnosing tuberculous pleurisy. Related data were extracted and sensitivity/specificity, positive/negative likelihood ratio (PLR/NLR), and diagnostic odds ratio (DOR) were pooled. Summary receiver operating characteristic curve and area under the curve (AUC) were performed and calculated to summarize the overall test performance. RESULTS: Fourteen studies involving 1382 subjects met inclusion criteria, including 715 cases of tuberculous pleurisy and 667 controls. Summary estimates of the diagnostic performance of the IP-10 for tuberculous pleurisy were listed as follows: sensitivity, 0.84 (95%CI 0.81 to 0.87); specificity, 0.90 (95% CI 0.88 to 0.92); PLR, 7.96 (95% CI 5.59 to 11.32); NLR, 0.19 (95% CI 0.15 to 0.24); DOR, 49.82 (95% CI 28.08 to 88.38); and AUC 0.94. No publication bias was detected. CONCLUSION: Pleural IP-10 is a useful diagnostic marker for tuberculous pleurisy. Nevertheless, its result should be interpreted together with the results of conventional test and clinical information of patients.

[Urothelial-type mucinous adenocarcinoma of the prostate: A case report and review of the literature].

OBJECTIVE: To investigate the clinical manifestations, pathological characteristics, and treatments of urothelial-type mucinous adenocarcinoma of the prostate (UMAP). METHODS: We reported a case of UMAP, reviewed relevant literature, and analyzed the clinicopaothological features, diagnosis, treatment, and prognosis of the disease. RESULTS: The patient was a 60-year-old male and underwent transurethral resection of the prostate for dysuria. Postoperative pathology indicated mucinous adenocarcinoma and sigmoidoscopy revealed no primary colon cancer. Immunohistochemical staining showed the negative expressions of PSA and P504s and positive expressions of CK7, CK34 ß E12, CK20, and CDX2. Thus UMAP was confirmed and treated by intensity-modulated radiotherapy. Then the patient was followed up for 30 months, which showed desirable therapeutic result, with neither local progression nor distant metastasis. CONCLUSION: UMAP has a bad prognosis and its diagnosis depends on pathological and immunohistocchemical examinations. It responds well to radical prostatectomy but is not sensitive to endocrine therapy. Radiotherapy can be considered for those who are not fit to receive radical prostatectomy.

BDNF-hypersecreting human umbilical cord blood mesenchymal stem cells promote erectile function in a rat model of cavernous nerve electrocautery injury.

PURPOSE: Erectile dysfunction (ED) continues to be a significant problem for men following radical prostatectomy. We hypothesize that intracavernous injection of BDNF-hypersecreting human umbilical cord blood mesenchymal stem cells (hUCB-MSCs) can ameliorate ED in a rat model of cavernous nerve electrocautery injury (CNEI). METHODS: Forty-two male Sprague-Dawley rats were randomly divided into four groups: sham + PBS (n = 6), CNEI + PBS (n = 12), CNEI + hUCB-MSCs (n = 12) and CNEI + BDNF-hUCB-MSCs (n = 12). At day 28 post-surgery, erectile function was examined and specimens were harvested for histology. Immunofluorescence staining, Masson's trichrome staining and transmission electron microscopy were performed to determine the structural changes in corpus cavernosum. Cells that are injected into penis were labeled by BrdU and tracked by immunofluorescence staining. Three days post-surgery, the concentration of BDNF protein in penile tissues was measured by Western blotting. RESULTS: Rats intracavernosally injected with BDNF-hUCB-MSCs showed the most significant improvement in the ratio of maximal ICP to MAP (ICP/MAP). Histological examinations showed moderate recovery of nNOS-positive nerve fibers, ratio of smooth muscle to collagen and smooth muscle content in the CNEI + hUCB-MSCs group and remarkable recovery in the CNEI + BDNF-hUCB-MSCs group compared to the CNEI + PBS group. By TEM examination, atrophy of myelinated and non-myelinated nerve fibers was noted in CNEI + PBS group and significant recovery was observed in two treated groups. There were more BrdU-positive cells in the BDNF-hUCB-MSCs group than in the hUCB-MSCs group both in the penis and in the MPG. Three days post-surgery, the concentration of BDNF protein in penile tissues in BDNF-hUCB-MSCs group was much higher than in other groups. CONCLUSIONS: Intracavernous injection of BDNF-hypersecreting hUCB-MSCs can enhance the recovery of erectile function, promote the CNs regeneration and inhibit corpus cavernosum fibrosis after CNEI in a rat model.

[Expression of ZEB1 in the prepuce of hypospadias children and its implication].

OBJECTIVE: To investigate the expression of zinc finger E-box binding homebox 1 (ZEB1) in the prepuce of hypospadias children and its relationship to the incidence of hypospadias. METHODS: Prepuce tissues were collected from 37 children aged 6-15 months undergoing hypospadias repair and 11 age-matched controls receiving circumcision. Based on the position of the urethral meatus, the hypospadias cases were classified as severe (n = 13) and mild-moderate (n = 24). The mRNA and protein expressions of ZEB1 were determined by immunohistochemistry and RT-PCR. RESULTS: The expression of the ZEB1 protein was remarkably higher in the severe (100% [13/13]) and mild-moderate hypospadias patients (75.0% [18/24]) than in the controls (9.1% [1/11]), with statistically significant differences between any two groups (P < 0.05). RT-PCR showed the integrated density value (IDV) of the ZEB1 mRNA expression to be (0.67 ± 0.21), (0.81 ± 0.24), and (1.55 ± 0.29) in the control, mild-moderate, and severe hypospadias patients, respectively, significantly higher in the severe hypospadias than in the control and mild-moderate hypospadias groups (P < 0.05), but with no significant difference between the latter two (P = 0.64). CONCLUSION: The expression of ZEB1 is significantly increased in hypospadias patients, and its upregulation is positively correlated with the severity of hypospadias, which suggests that the overexpression of ZEB1 may contribute to the development of hypospadias.

Therapeutic potential of human umbilical cord blood mesenchymal stem cells on erectile function in rats with cavernous nerve injury.

OBJECTIVE: To evaluate the therapeutic potential of human umbilical cord blood mesenchymal stem cells (hUCBMSCs) on promoting erectile function in a rat model of bilateral cavernous nerve (CN) crush injury. RESULTS: Fifty male Sprague-Dawley rats were randomly assigned to sham + PBS group (n = 10), BCNI (bilateral cavernous nerve crush injury) + PBS group (n = 10), BCNI + hUCBMSCs group (n = 30). At day 28 (n = 10) post-surgery, erectile function was examined and histological specimens were harvested. Compared with BCNI + PBS group, hUCBMSC intracavernous injection treatment significantly increased the mean ratio of ICP/MAP, nNOS-positive nerve fibers in the dorsal penile nerve, smooth muscle content, and smooth muscle to collagen ratio in the corpus cavernousum. Electron microscopy revealed few CN and major pelvic ganglion (MPG) lesions in the BCNI + hUCBMSCs group. Injected hUCBMSCs were localized to the sinusoid endothelium of the penis and MPG on day 1, 3, 7, and 28 post-intracavernous injection. CONCLUSION: hUCBMSCs intracavernous injection treatment improves erectile function by inhibiting corpus cavernosum fibrosis and exerting neuroregenerative effects on cell bodies of injured nerves at MPG in a BCNI rat model.

[Transurethral seminal vesiculoscopy combined with finasteride for recurrent hematospermia].

OBJECTIVE: To evaluate the clinical effectiveness of transurethral seminal vesiculoscopy (TUSV) combined with finasteride in the treatment of recurrent hemospermia. METHODS: This study included 32 patients with recurrent hematospermia, with the disease course of 3 months to 4 years. After administration of finasteride at 5 mg/d for 2 weeks, the patients underwent TUSV for both exploration of the causes and treatment, followed by medication with finasteride at the same dose for another 2 weeks. Postoperative follow-up was conducted for observation of the outcomes and complications. RESULTS: TUSV was successfully accomplished in all the 32 cases, which revealed 16 cases of seminal vesiculitis, 10 seminal calculi, 1 seminal vesicle cyst, 2 seminal vesicle polyps, and 3 seminal vesicle abscess. The operative time was 20 to 51 (31.0 +/- 5.2) minutes. Postoperative complications included 1 case of acute epididymitis and 3 cases of breast discomfort within the first 4 weeks. No incontinence, urethral stricture, rectal injury, retrograde ejaculation, and sexual dysfunction occurred postoperatively. All the patients but 1 were followed up for 6 months to 2 years. Twenty-nine of the cases were cured, and 2 experienced recurrence. CONCLUSION: Transurethral seminal vesiculoscopy combined with finasteride is safe and effective for the treatment of recurrent hemospermia.

Electrocautery-induced cavernous nerve injury in rats that mimics radical prostatectomy in humans.

OBJECTIVE: To investigate the early and delayed effects of cavernous nerve electrocautery injury (CNEI) in a rat model, with the expectation that this model could be used to test rehabilitation therapies for erectile dysfunction (ED) after radical prostatectomy (RP). MATERIALS AND METHODS: In all, 30 male Sprague-Dawley rats were randomly divided equally into two groups (15 per group). The control group received CNs exposure surgery only and the experimental group received bilateral CNEI. At 1, 4 and 16 weeks after surgery (five rats at each time point), the ratio of maximal intracavernosal pressure (ICP) to mean arterial pressure (MAP) was measured in the two groups. Neurofilament expression in the dorsal penile nerves was assessed by immunofluorescent staining and Masson's trichrome staining was used to assess the smooth muscle to collagen ratio in both groups. RESULTS: At the 1-week follow-up, the mean ICP/MAP was significantly lower in the CNEI group compared with the control group, at 9.94% vs 70.06% (P < 0.05). The mean ICP/MAP in the CNEI group was substantially increased at the 4- (35.97%) and 16-week (37.11%) follow-ups compared with the 1-week follow-up (P < 0.05). At all three follow-up time points, the CNEI group had significantly decreased neurofilament staining compared with the control group (P < 0.05). Also, neurofilament expressions in the CNEI group at both 4 and 16 weeks were significantly higher than that at 1 week (P < 0.05), but there was no difference between 4 and 16 weeks (P > 0.05). The smooth muscle to collagen ratio in the CNEI group was significantly lower than in the control group at the 4- and 16-week follow-ups (P < 0.05), and the ratio at 16 weeks was further reduced compared with that at 4 weeks (P < 0.05). CONCLUSIONS: In the CNEI rat model, we found the damaging effects of CNEI were accompanied by a decline in ICP, reduced numbers of nerve fibres in the dorsal penile nerve, and exacerbated fibrosis in the corpus cavernosum. This may provide a basis for studying potential preventative measures or treatment strategies to ameliorate ED caused by CNEI during RP.

[Semen quality and sperm ultrastructure in infertile men with varicocele].

OBJECTIVE: To examine and analyze semen quality and sperm ultrastructural characteristics of infertile patients with varicocele. METHODS: This study included 118 infertile patients with varicocele (the VC group) and 76 normal semen donors (the control group). We obtained routine semen parameters, seminal plasma biochemical markers and the levels of reproductive hormones in the subjects, and observed the changes in sperm structure under the scanning electron microscope and transmission electron microscope. RESULTS: Compared with the normal control, the VC patients showed significantly decreased sperm concentration, sperm progressive motility, sperm viability (P < 0.05), but no remarkable difference in semen volume and non-progressive motility (P > 0.05). The concentrations of zinc and alpha-glycoside enzyme in the seminal plasma were markedly reduced in the VC group in comparison with the controls (P < 0.05), but there was no significant difference in the level of fructose (P > 0.05), nor in such seminal plasma biochemical markers as FSH, LH, T and E2 between the two groups (P > 0.05). The percentage of morphologically normal sperm was dramatically lower in the VC than in the control group ([56.76 +/- 15.32]% vs [12.34 +/- 6.58]%, P < 0.05), and the sperm deformities were mostly in the head and neck, mainly tapering pin head accompanied by complex abnormal differentiation. CONCLUSION: This study demonstrated that VC may lead to oligo-astheno-terato zoospermia, and hence male infertility, which may be attributed to the changes of seminal plasma microenvironment and sperm ultrastructure.

Continuous treatment with new agents for newly diagnosed multiple myeloma.

To examine the role of novel agents such as bortezomib, lenalidomide, and thalidomide as continuous therapy (induction and consolidation/maintenance) in the treatment of newly diagnosed patients with multiple myeloma, we carried out a meta-analysis of randomized-controlled trials. A comprehensive literature search (Medline, Embase, the Cochrane controlled trials register, and the Science Citation Index) was performed. The initial search yielded 849 citations, of which 11 randomized-controlled trials enrolling 4775 patients fulfilled the inclusion criteria. Continuous addition of bortezomib to conventional therapy before and after autologous stem cell transplantation prolonged overall survival significantly: the summary hazard ratio was 0.80, 95% confidence interval [0.64, 0.99] (P=0.04). Continuous therapy with novel agents consistently improved progression-free survival (PFS) compared with therapy with conventional agents alone. For those patients ineligible for a transplant, the summary hazard ratios for PFS were 0.69 [0.56, 0.85] (P<0.001) for continuous thalidomide therapy and 0.47 [0.33, 0.68] (P<0.001) for continuous lenalidomide therapy; for those patients ineligible for a transplant, the summary hazard ratios for PFS were 0.68 [0.59, 0.79] (P<0.001) for continuous thalidomide therapy and 0.72 [0.61, 0.85] (P<0.001) for continuous lenalidomide therapy. In summary, continuous therapy with novel agents improved PFS consistently, and bortezomib may improve the overall survival of patients with newly diagnosed myeloma when it is added to standard transplantation therapy continuously.

[Stem cell therapy for erectile dysfunction].

Erectile dysfunction (ED), as a pathological phenomenon, refers to repeated or sustained difficulty to achieve and maintain sufficient penile erection to complete satisfactory sexual intercourse or sexual activity in male. The erectile reflex interruption induced by cavernous nerve (CN) damage is a direct cause of ED. In addition, the apoptosis of smooth muscle cells and endothelial cells in the corpus cavernosum caused by CN injury, along with the reduction of corpus cavernosum smooth muscle fibers, can increase the incidence of ED. Therefore, early intervention of the pathological process of CN injury and promotion of CN regeneration are essential for the treatment of ED. In recent years, the stem cell therapy for ED has become a focus in clinical research. This article offers an overview on the application of embryonic stem cells, mesenchymal stem cells, muscle-derived stem cells, and adipose stem cells in the treatment of ED.

[Differential gene expressions in the stromal cells of the human prostate peripheral zone in males of different ages].

OBJECTIVE: To study the gene expressions in the stromal cells of the human prostate peripheral zone (PZ) in men of different ages. METHODS: We primarily cultured stromal cells from the normal prostate PZ of men aged 23 -32 (young group) and 56 -75 years (old group), profiled the gene signature of the PZ cells by cDNA microarray, and defined the differential gene expression patterns by hierarchical cluster analysis. Among the differential genes, we selected and confirmed up-regulated genes by quantitative real time PCR (Q-PCR), and identified their protein coding by Western blotting. RESULTS: There were significant differences in the gene expressions of the PZ cells between the old and young groups. Based on the fold change ratio of > or = 2 or < or = 0.5, 509 up-regulated and 188 down-regulated genes were selected in the PZ cells. A subset of significantly differential genes influencing the growth of adjacent epithelial cells were identified, including HGF, IGF2, IGFBP5 and MMP1 in the old males. CONCLUSION: Stromal cells in the prostate PZ were more active in older males in promoting the malignant progression of adjacent prostate epithelial cells, which might be due to the increased expression of extracellular paracrining mediators.

[Compound graft of porcine small intestinal submucosa with Schwann cells to reconstruct injured cavernous nerves and restore erectile function].

OBJECTIVE: To investigate the restoration of rat penile erection by reconstructing injured cavernous nerves (CN) with a compound graft prepared from porcine small intestinal submucosa (SIS) and Schwann cells (SC). METHODS: SCs were cultured in vitro and a compound graft was prepared from the SCs and SIS. Thirty-three healthy SD rats were randomly divided into three groups of equal number, sham-operation, CN ablation, and SIS + SC graft. Three months after the operation, all the rats underwent the apomorphine test, followed by immunohistochemical staining of the tissues from the middle part of the corpus cavernosum penis. RESULTS: Combined use of mechanical stripping, mixed-enzyme digestion, different-speed adhesion, short-term Ara-C and some other methods yielded SCs of a purity high enough for nerve tissue engineering. The SIS prepared by mechanical and chemical methods exhibited a good biocompatibility with SCs, which could adhere, grow, propagate and differentiate on its surface. The apomorphine test showed that both the rate and frequency of penile erection were significantly higher in the SIS + SC graft than in the CN ablation group (P < 0.01), but lower than in the sham operation group (P < 0.01). The number of nNOS positive nerve fibers in the SIS + SC graft group was significantly different from that of the CN ablation (P < 0.01), but both were smaller than that of the sham-operation group. CONCLUSION: The compound of SIS with SCs, as a nerve graft, can be used to reconstruct injured cavernous nerves, and to some extent, restore penile erectile function.

[Restoration of erectile function by reconstructing cavernous nerves with small intestinal submucosa grafts].

OBJECTIVE: To investigate the restoration of erectile function by reconstructing cavernous nerves (CN) with small intestinal submucosa (SIS) grafts. METHODS: We prepared SIS grafts, established rat models and divided the models into a CN ablation, a sham-operation and an SIS graft group. The CNs at both sides were severed with 1 cm ablated in the first group, and 0.5 cm removed in the third, followed by reconstruction with the SIS grafts. Three months after surgery, the apomorphine test was performed to evaluate the erectile function, and then all the rats were sacrificed to detect the expression of nNOS in the penis. RESULTS: Penile erection was observed in 72.73% (8/11) of the rats for (1.07 +/- 0.89) times within 30 min in the SIS graft group, as compared with 0% (0/11) of the rats for (0.00 +/- 0.00) times in the CN ablation group (P < 0.01), and 90.91% (10/11) of the rats for (2.19 +/- 1.17) times in the sham-operation group (P < 0.01). The number of nNOS nerve fibers was significantly larger in the SIS graft than in the CN ablation group (70.36 +/- 10.09 versus 22.09 +/- 4.76, P < 0.01), but both were significantly smaller than that of the sham-operation group (90.81 +/- 5.69, P < 0.01). CONCLUSION: The SIS grafting technique contributes to the recanalization of the severed CN and restoration of erectile function in rats after surgical injury.

Cadaveric study of nerves supplying the membranous urethra.

AIMS: To better understand the anatomy of the region of the male membranous urethra in order to preserve continence during radical cystectomy and prostatectomy. METHODS: Cadaveric dissections of 15 male specimens were undertaken to investigate the nerves to membranous urethra. The nerves were traced from both an intrapelvic approach and a perineal approach. The origin, course, and distribution of the branches to the membranous urethra region were investigated in detail. RESULTS: The membranous urethra is innervated by branches of inferior hypogastric plexus (IHP) and intrapelvic and extrapelvic branches of pudendal nerve (PN). The pelvic nerve from IHP originated from the caudal most root of the pelvic splanchnic nerve, running along the surface of the levator ani muscle (LAM) to enter the membranous urethra at the 5 and 7 O'clock positions. In 40% of specimens we found that the intrapelvic branches were supplied by the PN. Before exiting the pudendal canal, PN gives off an intrapelvic branch that traverses the LAM to course with the pelvic nerve and innervate the membranous urethra; the distance between these intrapelvic branches and prostatic apex is 5.3 +/- 1.8 mm. The branches originating from the dorsal nerve of penis innervate the membranous urethra in 53.3% of specimens; these nerve branches are located 4.2 +/- 1.1 mm from the prostatic apex. CONCLUSIONS: Dissection of the seminal vesicles and the prostatic apex during radical cystectomy and prostatectomy likely injures the nerve responsible for continence.

[Reconstruction and repair of resected cavernous nerves].

By studying the novel methods for reconstructing damaged cavernous nerves and the related literature on the regeneration of cavernous nerves, restoration of erectile function and neurohistological reconstruction engineering, a variety of grafting materials have been found applicable to cavernous nerve reconstruction, including autogenetic nerve grafts, silicone tubes, artificial biodegradable conduits and so on. Neurotrophic factors, extra cellular matrix components and Schwann cells have been shown to promote cavernous regeneration. Artificial nerve guides, especially biodegradable ones containing growth-promoting factors or cells, are a promising option for the repair of cavernous nerve lesions.