Object To establish a calli culture system for the rapid propagation of Speiraea japonica L. f.. Methods Callus and shoot induction were carried out on MS, 6, 7 V or B 5 media with different parts of the plant such as stem tip, tender leaves and petiole as explants. Results A calli culture system was established for the rapid propagation of S. japonica. Conclusion MS cultural medium was found to be most suitable for calli induction. MS with 2.0 mg/L 2,4 D+0.3 mg/L KT can induce calli when the explants were used for the induction, with stem tips being the most satisfactory explant. Clusters of seedlings can be induced on MS+2 mg/L BA+0.1 mg/L NAA and when these seedlings were transferred to 1/2 MS +0.25 mg/L BA+0.5 mg/L NAA medium, root were developed to give young seedlings.

BACKGROUND:There is few mesenchymal stem cells(MSCs) in the bone marrow,about one BMSC in 1?104-1?105 monocytes.Following in vitro isolation,purification and amplification,it is satisfactory for in vivo requirement.OBJECTIVE:To observe biological characteristics and potentiality of multi-directional differentiation of BMSCs in vitro.DESIGN,TIME AND SETTING:The cytological in vitro study was performed at the Institute of Urinary Surgery,Union Hospital Affiliated to Fujian Medical University from October 2007 to December 2008.MATERIALS:A total of 30 clean male Sprague Dawley rats of inbreeding line were supplied by Silaike,Shanghai,China.METHODS:BMSCs from rats were isolated and cultured in vitro by using the whole bone marrow adherence method.When 80%-90% confluency,BMSCs were digested by trypsin.At the third passage,BMSCs were induced to differentiate into osteoblasts and adipocytes.MAIN OUTCOME MEASURES:Cell morphology was observed under the inverted phase contrast microscope.BMSCs surface markers were detected using flow cytometry.Osteogenic ability was examined by alizarin red staining.Adipogenic ability was measured by Oil red O staining.RESULTS:The primary cells and the passage cells were mostly fusiform in shape,to be similar to fibroblasts.Cell still kept a high potential of growth and amplification following over 10 subcultures.At the third passage,BMSCs were positive for CD44,CD90,CD106,but negative for CD34,CD45,CD11b.Following 21 days of osteogenic induction,cell alizarin red staining showed that alizarin red was positive in osteoblasts.Following 2 weeks of adipogenic induction,oil red O staining showed that red lipid droplet existed in adipocytes.CONCLUSION:Whole bone marrow adherence method can isolate,purify and amplify BMSCs in vitro.The obtained cells have general biological characteristics of MSCs,and also have potentiality of multi-directional differentiation.

Objective To confirm that rat bone marrow mesenchymal stem cells (MSC) transfected with nerve growth factor (NGF) gene in the bladder tissue of diabetic rats bladder tissues can survive and stably express NGF. Methods A diabetic rat model was constructed. The BrdU-labelled MSC transfected with NGF gene were transplanted into the diabetic rats bladder tissues. BrdUlabelled immunohistochemistry was used to observe the growth of MSC transfected with NGF gene in the diabetic rats bladder tissues. The expression of NGF mRNA and protein were checked by RT-PCR and ELISA. Results A diabetic rat model was successfully built by a single intraperitoneal injectionof STZ. The blood glucose was still high after 8 weeks. NGF gene modified MSC could be detected in the bladder of diabetic rats by BrdU-labelled immunohistochemistry. The concentration of NGF in the control group, disease group and treatment group were ( 114 ± 3), ( 70 ± 2), ( 110 ± 2) pg/ml by ELISA and mRNA quantity by RT-PCR were 0. 183±0. 004, 0. 032±0. 139, 0. 130±0. 165, respectively. Compared with the control group, the expression of NGF gene was decreased (P＜0. 05) in the incidence group. The expression of NGF gene was increased (P＜0. 05) in the treatment group compared with the disease group. Conclusions The NGF gene-modified MSC could survive in diabetic rats bladder tissues. The NGF gene in MSC could stably express in diabetic rats bladder tissues.

Objective To assess the causes of sildenafil failure and the feasibility of successfully rechallenging non-responding patients. Methods A total of 126 consecutive erectile dysfunction (ED) patients from Jan 2009 to Dec 2009 who claimed poor response to sildenafil (sildenafil 100 mg on demand, at lease 4 epiodes) were enrolled into the study. All patients received sexual reeducation and were treated with sildenafil, taken on a daily dose of 50 mg for 4 weeks. The International Index of Erectile Function-5 (IIEF-5), Rigiscan, serum testosterone or penile Doppler were used to evaluate ED and the cause of on-demand sildenafil failure. End point efficacy of rechallenging was evaluated using the IIEF-5 and the sexual encounter profile (SEP) 'Were you able to insert your penis into your partner's vagina?' and 'Did your erection last long enough to achieve successful intercourse?'. Results The recruited patients comprised of 41 cases with psychological ED, 39 cases with hypogonadism ED, 28 cases with diabetes mellitus ED and 18 cases with vascular ED. Compared with pretreatment and on-demand sildenafil baseline, daily administration of sildenafil significantly enhanced all efficacy outcome variables. The IIEF-5 was significantly improved after daily sildenafil (12.3 ± 2.9 vs18. 8±4.4, P＜0.01), 78 patients responded to daily sildenafil. The overall salvage rate was 61.9%(78/126). Conclusions Sexual reeducation and daily administration of sildenafil may be able to salvage many patients with ED who were sildenafil non-responders.

Objective To estabalish a new conception, Erectile Dysfunction-no sexual life (ED-NS), so as create an evaluating questionnaire, Self-estimation Index of erectile function-No sexual life (SIEF-NS) so as to investigate its clinical reliability. Methods The conception of ED-NS was identified and the SIEF-NS questionnaire was established. Patients who complained of ED-NS and normal controls were enrolled into the research and assessed the erectile function was assessed with SIEF-NS.The SIEF-NS includes 12 questions, such as sexual libido, general erectile function, nocturnal penile erection, erectile function during foreplay, erectile function during audio-video sexual stimulation,confidence, depression, etc and each question has 5 point scales. Results Sixty-one ED-NS patients and 57 controls were enrolled into the study and assessed erectile function with SIEF-NS. The mean score of each question and integral score of SIEF-NS in ED-NS patients were significantly different from normal controls (P＜0. 05). When the integral score was 35 points according to the ROC curve of integral score, the sensitivity of SIEF-NS was 88.5% and specificity was 96. 5%. Conclusions ED-NS is a new conception to define patients who have erectile dysfunction without sexual life. SIEFNS is suggested to be a useful method for the evaluation of ED-NS patients.

To study the chemical constituents of Lysimachia patungensis Hand.-Mazz., silica gel column chromatography, reverse phase ODS column chromatography, MCI and Sephadex LH-20, were used to separate the 95% EtOH extract of the whole plant of Lysimachia patungensis Hand.-Mazz.. The structures of the isolated compounds have been established on the basis of chemical and NMR spectroscopic evidence as well as ESI-MS in some cases. Twelve phenolic compounds were obtained and identified as quercetin-3, 3'-di- O-alpha-L-rhamnoside (1), myricetrin (2), quercitrin (3), rutin (4), 2-hydroxynaringenin-4'-O-glucopyranoside (5), naringenin 7-O-glucopyranoside (6), liquiritin apioside (7), licochalcone B (8), tetrahydroxymethoxy chalcone (9), methyl-p-coumarate (10), 2, 4, 6-trihydroxy acetophenone-2-O-glucopyranoside (11) and vaccihein A (12). Among them, compound 1 is a new compound, and compounds 5, 11 and 12 are isolated from the genus Lysimachia L. for the first time, and the others are isolated from the plant for the first time.

Objective To evaluate the efficacy of microvasoepididymostomy for the treatment of obstructive azoospermia. Methods Ninety-eight patients were confirmed as obstructive azoospermia by infertility investigations.The mean (range) age was 31 (20-43) years.The mean duration of obstruetion was 4 years.All patients were seen by the surgeon for a complete history and physical examination.Semen analyses proved azoopermia,serum levels of sexual hormone were normal and testicular function for production of sperms were normal certified by biopsy.All patients with suspected epididymal or vasal obstruction were offered scrotal exploration.The decision for microvasoepididymostomy was made during surgery,based on the pateney of the vas towards the abdomen documented by saline and sperm fragments containing in the epididymal fluid on intraoperative light-microscopic examination.Microvasoepididymostomy was performed in 58 patients documented epididymal obstruction The initial semen analysis was then done after usually 3 months postoperatively.Patency was arbitrarily defined as＞10 000 sperm/mL in ejaculate in at least one semen analysis after surgery.Postoperative patency rate and postoperative impregnation rate were followed. Results Fifty patients were followed up for 3-29 months,8 cases lost.Sperm was found by semen analysis in 36 patients.Sperm density was 4×104-2×108 sperms/ml and motility was 2%-70%.Semen analysis reveals azoospermia in 6 patients and the patients were counseled to undergo further testing to determine the ultimate outcome of the procedure.Natural conception occurred in 14 patients followed for more than 12 months.The overall pateney rate was 72%(36/50).Among patients with a follow-up of＞6 months,the natural paternity rate was 28%(14/50).The median time to achieve a natural pregnancy was 6.6 (4.0-10.0)months. Conclusion A more favourable patency can be achieved using microsurgical intussusception vasoepididymostomy and a part of patients with obstructive azoospermia can be cured by this means.

Objective To compare the circumferential approach and suprapubic approach for correcting the concealed penis. Methods Thirty-four patients received circumferential approach (group A,18 cases)or suprapubic approach(group B,15 cases)randomly for the concealed penis.The length of penis without erection before and after surgery treatment,patients'satisfaction and surgical complications were evaluated.All the patients were followed at least 6 months after operation.Results The penile length before and after surgery in group A were(1.78±0.41)cm vs(3.97±0.47)cm,the length of the postoperative penis was significantly longer than that of the preoperative penis(P<0.01).83%of those in group A(15 cases)were satisfied with the cosmic results.The length of preoperative and postoperative penis in group B were(2.07±0.53)cm vs(4.05±0.81)cm respectively,the length of the postoperative penis was significantly longer(P<0.01).Satisfaction was 87%(13 cases).The postoperative penile length between the 2 groups was not different(P> 0.05).Prepuce edema was reported to Occur in 56%and 13%in group A and group B and spontaneously disappeared within 3 months.Fat synchysis occurred in 2 of group B. Conclusions The cosmic effect of the 2 surgical approaches is similar,but circumferential approach is simpler.There is no serious complication in those receiving circumferential approach.

Objective To review the clinical manifestation,pathological characteristics,treat ment and prognosis of malignant mesothelioma of the tuniea vaginalis testis.Methods A case of ma lignant mesothelioma of the tunica vaginalis testis was reported and relative literature was reviewed.A34 year old man was admitted to hospital with a history of aggravated hydroeele for 5 years.A hardtumor was palpated above the right testis,which was smooth and slight tenderness.The level of β-hCG and AFP was normal.Ultrasonography revealed a 2.05 cm× 1.97 cm,well demarcated,mixedecho tumor.Results An operation was performed to incise the tumor and tunica vaginalis undersubarachnoid space block anesthesia.The tumor was histologically diagnosed as low potential malig nant mesothelioma of the tunica vaginalis testis.Microscopic examination showed that the tumor wasmalignant with a biphasic pattern composed of epithelial and stromal components.The former compo nent was dominant.The epithelial cells proliferated obviously with atypia and mitosis.Immunohisto chemical staining showed cytokeratin and vimentin were strongly positive.The capillary,lymph ves sel,spermatic cord and testis were not infiltrated.Right radical orchiectomy and right scrotectomywas performed one month after the first operation.There was no evidence of relapse during 38 monthfollow up after operation.Conclusions The major symptom of patients with malignant mesotheliomais hydrocele,malignant mesothelioma is diagnosed by histopathology.Due to the invasive potential ofthis disease and the risk of tumor recurrence,radical orchiectomy and close follow up are strongly rec ommended.

Objective To investigate the expressions of fatty acid-binding protein 5 (FABP5)and dihydroli-poamide dehydrogenase(DLD)in skin lesions of symmetrical acrokeratoderma(SAK), and to explore their significance. Methods Biopsy specimens were obtained from skin lesions on the wrists and perilesional skin of 9 patients with SAK, and from normal skin in the wrists of 9 healthy volunteers (control group). Reverse transcription PCR (RT-PCR)and immunohistochemical staining were performed to measure the expressions of FABP5 and DLD in these specimens. Results RT-PCR showed no significant differences in the mRNA expressions of FABP5 or DLD between lesional, perilesional and normal control skin specimens(both P > 0.05). Immunohistochemically, there was a significant increase in the extent and intensity of staining for FABP5 in SAK lesions. Concretely speaking, FABP5 was strongly expressed in the stratum corneum, granular and spinous layers in SAK lesions, but weakly expressed in the stratum corneum, granular and spinous layers in perilesional skin, and only in spinous and basal layers in normal control skin. The expression of DLD decreased in SAK lesions, and was observed only in the stratum corneum and spinous layer in a few cases of SAK. However, the full-thickness epidermis stained positive for DLD in perilesional skin, with the nuclei and cytoplasm both stained deep brown. Conclusion The overexpression of FABP5 in SAK lesions may participate in dysdifferentiation of keratinocytes, while the down-regulation of DLD expression suggests an imbalance in energy metabolism.