The single needle method for urethrovesical anastomosis with strengthened posterior fixation during laparoscopic radical prostatectomy was explored. The method was initiated by performing a fixing suture with a knot at 4 o'clock of the posterior lip of bladder neck, and another suture at nearby position was performed to leave the knot outside. From 5 o'clock to 8 o'clock, sutures were performed every one o'clock to secure posterior approximation, then every two o'clock a suture. To avoid a loose anastomosis, lock sutures were performed every 3 sutures. The needle was always driven full-thickness outside-in in the bladder neck and inside-out on the urethra. After completing the full circumference, the needle was drawn near the 4 o'clock and tied at the tail end. Any leakage could be closed with additional interrupted sutures. The clinical data of 89 patients who underwent this method were retrospectively compared with those of 23 patients who underwent the single knot method. The results showed that the anastomosis, operative and catheterization time was 17.6+/-4.7 min, 134.0+/-10.7 min and 6.5+1.6 days respectively. There were 3 temporal urinary leakages identified in 89 cases requiring prolonged catheterization. No urinary leak and anastomotic stricture was confirmed, and 95.2% patients had total urinary control. It was concluded that this method was simple and safe for urethrovesical anastomosis.

Objective To describe the single needle running suture method for the urethrovesi-cal anastomosis during laparoscopic radical prostatectomy(LRP). Methods Forty-five patients of prostate cancer underwent LRP with the single needle running suture method. The technique was initi-ated by performing a fixing suture at the posterior lip of bladder neck at 4 o' clock and tying the first knot. Another suture at the nearby position of the first suture was performed to leave the first knot outside. From 5 o' clock to 8 o' clock, sutures were performed every one o' clock to secure posterior approximation, then every two o'clock a suture. To avoid a loose anastomosis, lock sutures were per-formed every 3 sutures. After completing the full circumference, the needle was drawn at the 2 o' clock for the second knot. The needle was always driven full-thickness outside-in in the bladder neck and inside-out on the urethra. Any remaining leakage could be closed with additional interrupted su-tures. Results All urethrovesical anastomosis were completed successfully. The mean anastomosis time was 16 rain(from 12 to 25 min), and mean operative time was 132 rain (112 to 185 rain). The mean catheterization time was 9 d(7 to 14 d). Three temporal urinary leaks requiring prolonged cathe-terization were identified. Forty-four patients had total urinary control in 1 year postoperatively and no other short-term or persistent complication was found with a mean follow-up of 21 months. Conclu- sion The single needle running suture method could be a simple and safe method for urethrovesical anastomosis during LRP.

OBJECTIVETo investigate the expression of microRNA-100 (miR-100) in human gastric cancer cells and its role of miR-100 in migration, invasion and proliferation in gastric cancer cell line SGC7901.

METHODSTotal RNAs were extracted from formalin-fixed and paraffin-embedded tissue samples of SGC7901 cells, gastric cancer (50 cases), non-tumor (18 cases) and lymph nodes with metastases (18 cases). The expression of miR-100 was examined by reverse transcription (RT)-qPCR. Additionally, SGC7901 cells were transfected with Pre-miR-100 and negative control constructs, and then their ability of migration, invasion and proliferation in vitro was documented after 48 hours.

RESULTSRT-qPCR showed that although miR-100 expressed in all samples, compared to non-tumour tissues, the expression was lower both in SGC7901 cells and gastric cancer tissues (P = 0.0077, P < 0.01). SGC7901 cells and primary gastric cancer tissues with lymph nodes metastasis had lower miR-100 expression than those of without lymph node metastasis (P = 0.0361, P = 0.0356). The migration ability and invasion of SGC7901 cells transfeced with pre-miR-100 decreased as compared with control cells (P = 0.0025, P = 0.0028 respectively). However, miR-100 expression had no significant effects on the cell proliferation.

CONCLUSIONSExpression of miR-100 inhibits the migration and invasion of gastric cancer cells without significant alteration of proliferation. Therefore, miR-100 may play an inhibitory role in the progression of gastric carcinoma.

Adult ; Aged ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Female ; Humans ; Lymphatic Metastasis ; Male ; MicroRNAs ; genetics ; metabolism ; Middle Aged ; Neoplasm Invasiveness ; Stomach Neoplasms ; genetics ; pathology ; Transfection

Objective To develop and evaluate a porcine model for training the single needle running suture method of laparoscopie urethrovesical anastomosis(LUA). Methods Twenty minipigs with mean weight of 30kg were general anaesthetized with Sumianxin solution 0. 1 ml/kg intramuscularly. Pneumoperitoneum was created by insufflation of carbon dioxide by a veress needle inserted through the umbilicus. One 10mm port and two 5mm ports were positioned after the establishment of pneumoperitoneum. The intestine was used as "bladder". The procedures were completed with the single needle running suture method of laparoscopic urethrovesical anastomosis. Six trainees performed the LUA procedure based on the models during a laparoscopic training course, following the technique used in the operation room. The learning curve was analyzed by operative time. Results The porcine model for laparoscopic training was established successfully and 3 LUAs could be performed on each pig. Each trainee performed 10 LUAs based on the models during the training course of laparoscopic urology. The operative time declined from (55.3±10. 4)min initially to (22.4±4.8)min (P＜0. 01) after the training course. At the end of training, all trainees could accomplish a watertight LUR procedure on the model. Conclusions The establishment of the training model is feasible. The trainees could acquire the skills necessary to perform LUA in vivo based on this model. The model provides a platform for training the basic techniques of LUA procedures.

The medicinal fungi, which are of great importance in traditional medicine, are facing the problems of wild resources scarcity and low concentration of bioactive compounds. Velvet family and LaeA global regulator play a vital role in secondary metabolism and developmental programs, which are found in a wide variety of fungi ranging from Chytridiomycota to Basidiomycota. This review elaborates the structures and functions between Velvet family and LaeA protein. The Velvet family which shares the Velvet protein domain, including VeA (Velvet), VelB (Velvet like B), VosA (viability of spores A) and VelC (Velvet like C), acts on the regulation function is secondary metabolism and developmental programs such as asexual and sexual development. Furthermore, the function is affected by environmental factors such as light and temperature. LaeA protein which owns S-adenosylmethionine-dependent methyltransferase domain, coordinately regulates development and secondary metabolism by regulating and modifying the Velvet proteins. The regulation of LaeA is mediated by light receptor proteins. Therefore, clarifying the mechanism of Velvet and LaeA proteins in medicinal fungi will pave the way for nurturing medicinal fungi and improving production of bioactive compounds.
Fungal Proteins ; metabolism ; Fungi ; chemistry ; Gene Expression Regulation, Fungal ; Genes, Regulator ; Protein Structure, Tertiary ; Secondary Metabolism ; Structure-Activity Relationship

OBJECTIVETo explore the relation between the expressions of PD-ECGF and VEGF and the evolution of capillary hemangioma, so as to provide theoretical basis for treatment.

METHODSFourty cases with capillary hemangioma, proved by pathologic method, were randomly selected and divided into proliferative (n=22) and involuted groups (n=18), according to the Mulliken standard. 8 specimens from 8 children with prepuce operation were used as control group. All the specimens were fixed, embedded and underwent HE staining. The expression of PD-ECGF, VEGF and CD34 in endothelial cells were detected by immunohistochemistry. The microvessel-density (MVD) was also calculated. The results were analyzed by SPSS12.0.

RESULTSThe positive expression rates of PD-ECGF and VEGF were 95.45% (21/22) and 86.36% (19/22) in proliferative hemangioma, 77.78% (14/18) and 66.67% (12/ 18) in involuted hemangioma, 37.50% (3/8) and 37.50% (3/8) in normal skin. MVD in proliferative and involuted hemangioma and normal skin was 93.68 +/- 20.56, 51.94 +/- 20.73 and 17.50 +/- 5.30, respectively. There was a significant difference in PD-ECGF expression and MVD between the proliferative and involuted groups, or between the hemangioma and control groups (P < 0.05). The VEGF was significantly different between the proliferative and involuted groups, or between the proliferative and control groups (P < 0.05), but not between the involuted and control groups (P > 0.05). The expression of VEGF, PD-ECGD and MVD showed a positive relationship.

CONCLUSIONSPD-ECGF and VEGF have a synergetic effect in the proliferation of micro-vessels. PD-ECGF may enhance the activity of thymidine phosphorylase. They play an important role in the proliferation and involution of hemangioma.

Child, Preschool ; Female ; Hemangioma, Capillary ; metabolism ; pathology ; Humans ; Infant ; Male ; Neoplastic Syndromes, Hereditary ; metabolism ; pathology ; Thymidine Phosphorylase ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

Transcription factor is one of the key factors in the regulation of gene expression at the transcriptional level. It plays an important role in plant growth, active components biosynthesis and response to environmental change. This paper summarized the structure and classification of bHLH transcription factors and elaborated the research progress of bHLH transcription factors which regulate the active components in plants, such as flavonoids, alkaloids, and terpenoids. In addition, the possibility of increasing the concentration of active substances by bHLH in medicinal plants was assessed. The paper emphasized great significance of model plants and multidisciplinary research fields including modern genomics, transcriptomics, metabolomics and bioinformatics, providing the contribution to improve the discovery and function characterization of bHLH transcription factors. Accelerating the research in the mechanism of bHLH transcription factors on the regulation of active components biosynthesis will promote the development of breeding and variety improvement of Chinese medicinal materials, also ease the pressure of resources exhaustion of traditional Chinese medicine home and abroad.
Alkaloids ; biosynthesis ; Basic Helix-Loop-Helix Transcription Factors ; chemistry ; classification ; genetics ; metabolism ; Flavonoids ; biosynthesis ; Plants, Medicinal ; genetics ; metabolism ; Terpenes ; metabolism

ObjectiveTo investigate the effects of over expression of miR-34a on cellular proliferation and migration in bladder cancer cell line J82 by targeting Notchl.MethodsmiR-34a was predicted as a putative gene which can target Notchl through bioinformatics analysis,qRT-PCR and Western blot were performed to measure the expression levels of Notchl and miR-34a in invasive transitional cell carcinoma of bladder (TCCB) tissues and J82 cells transfected with miR-34a.Luciferase assay was employed to determine if miR-34a could target Notchl through binding to the 3'-untranslated region (3'UTR) of Notchl mRNA.J82 cells were transfected with pcDNA3.0-miR-34a or pcDNA3.0 control plasmid.MTS colorimetry was used to evaluate the effect of miR-34a on cell proliferation.The effect of miR-34a on cell migration was assessed by transwell migration assay.ResultsThe expression level of miR-34 in invasive TCCB tissues was lower than in adjacent bladder tissues (0.016(0.018) vs 0.042 (0.059),N =16; P =0.0006).On the contrary,the average levels of Notchl mRNA and protein were higher in tumors than in adjacent bladder tissues (2.765(2.156) vs 2.312(1.365),N =16; P =0.0025 and 0.857 ±0.197 vs 0.648 ±0.171 ;P ＜0.0001 ).After the transfection of miR-34a,the expressive level of miR-34a in J82 was highly induced ( (2.408 ±0.789) × 10-4 vs(0.153 ±0.029) × 10-4; P =0.0026).However,the expressive levels of Notchl mRNA and protein were obviously decreased (3.001 ± 0.106 vs 4.998 ± 1.053 ; P =0.0308 and 0.747 ± 0.050 vs 0.988 ± 0.102 ; P =0.0215 ).The results of luciferase assay showed that firefly activity was highly dimished (0.422 ± 0.028 vs 2.392 ± 0.148 ; P ＜ 0.0001 ).Cellular proliferation was inhibited after the transfection of miR-34a in J82 (P ＜ 0.0001 ).Moreover,number of migration cells of J82 was significantly reduced after the ectopic expression of miR-34a ( 179.3 ± 21.02 vs 269.7 ± 23.71 ; P =0.0078 ).ConclusionsmiR-34a inhibits the cellular proliferation and migration of bladder cancer cell line J82 via binding to the 3UTR of Notchl mRNA.

To investigate the relationship between the expression of RASSF1A protein and promoter hypermethylation of RASSF1A gene, RASSF1A protein expression was measured by Western blotting in 10 specimens of normal bladder tissues and 23 specimens of bladder transitional cell carcinoma (BTCC). The promoter methylation in BTCC and normal bladder tissues was detected by methylation-specific PCR (MSP). The results showed that the expression level of RASSF1A protein was significantly lower in BTCC tissues than that in normal bladder tissues. However, it was not correlated with its clinical stages and pathological grades. The frequency of promoter methylation of RASSF1A gene was higher in BTCC tissues than that in normal bladder tissues. In 14 patients with the aberrant promoter methylation, 13 showed loss or low expression of RASSF1A protein. It is concluded that RASSF1A gene promoter methylation may contribute to the low level or loss of RASSF1A protein expression, the inactivation of RASSF1A gene and the genesis of BTCC. But, it may bear no correlation with its clinical stages and pathological grades.
Blotting, Western ; Carcinoma, Transitional Cell/metabolism ; DNA Methylation ; DNA Primers/chemistry ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; Promoter Regions, Genetic ; Tumor Suppressor Proteins/*biosynthesis ; Tumor Suppressor Proteins/*genetics ; Urinary Bladder Neoplasms/*metabolism

ObjectiveTo analyze characteristic changes of shoulder muscles by investigating surface electromyographic regularity changes before and after the treatment of Frozen shoulders. Methods7 cases of frozen shoulders were selected, surface electromyography (sEMG) before and after treatment were recorded by AMT-8 EMG recorder. The collected data was processed using MATLAB software integrated EMG (IEMG) value were obtained. Then the IEMG data were statistically analyzed using Stata11.0 software and compared. Results ①The differences of abduction deltoid and infraspinatus muscle IEMG values before and after treatment were statistically significant(P ＜0.05), while IEMG values changes of biceps, triceps, pectoralis major, latissimus dorsi and trapezius before and after treatment were not statistically significant (P＞0.05); changes of adduction IEMG values of all muscles were not statistically significant (P＞0.05). ②Changes of flexion IEMG values of biceps before and after treatment were statistically significant (P＜0.05), while IEMG values changes of pectoralis major, infraspinatus muscle, latissimus dorsi, trapezius, deltoid, triceps before and after treatment were not statistically significant (P＞0.05). Changes of extension IEMG values of triceps before and after treatment were statistically significant (P＜0.05), while those of pectoralis major, infraspinatus muscle, latissimus dorsi, trapezius, deltoid and biceps were not statistically significant (P＞0.05). ③IEMG values of all the muscles during external rotation, internal rotation before and after treatment IEMG were not significantly different. Conclusion①After treatment, the outreach functions of infraspinatus and deltoid muscles were improved, while adduction functions of all muscles were not improved. ②Flexion function of biceps was improved significantly after treatment as well as extension function of triceps.③Extemal rotation, internal rotation functions of all muscles were not improved significantly.