Comparison of embryo implantation potential between time-lapse incubators and standard incubators: a randomized controlled study.

RESEARCH QUESTION: What are the potential clinical benefits of embryo culture and assessment in a time-lapse incubator compared with a standard incubator using static assessment? DESIGN: This large multicentre, single-blinded, randomized controlled study included 1224 participants randomly assigned (1:1) to the time-lapse or standard incubator group. In all patients one or two embryos were transferred on day 3. The primary outcome was the implantation rate in the first embryo transfer cycle. Secondary outcomes included the cumulative implantation rate, live birth rate in the first embryo transfer cycle and cumulative live birth rate. RESULTS: Among 1224 participants recruited, 1182 underwent embryo transfer. The number of successfully implanted embryos in the first transfer cycle was significantly higher in the time-lapse incubator group (time-lapse group: 52.35%, standard incubator group: 47.11%, P = 0.014). The implantation rate in the first embryo transfer cycle was still significantly higher in the time-lapse group than the standard incubator group after adjusting for age, body mass index, medical centre and embryo status (relative risk 1.11, 95% confidence interval 1.02-1.20, P = 0.020). However, the cumulative implantation rate, live birth rate in the first embryo transfer cycle and cumulative live birth rate were not statistically different between the groups. CONCLUSIONS: The implantation rate in the first embryo transfer cycle was significantly improved in the time-lapse group, but the effect of the time-lapse system on the cumulative implantation rate or cumulative live birth rate was not significant. The embryo assessment method offered by time-lapse systems rather than an undisturbed environment may play an important role in improving the implantation rate in the first embryo transfer cycle. These results are only applicable to young patients.

Nuclear receptor coactivator 6 promotes HTR-8/SVneo cell invasion and migration by activating NF-κB-mediated MMP9 transcription.

OBJECTIVES: NCOA6 is a transcription coactivator; its deletion in mice results in growth retardation and lethality between 8.5 and 12.5 dpc with defects in the placenta. However, the transcription factor(s) and the mechanism(s) involved in the function of NCOA6 in placentation have not been elucidated. Here, the roles of NCOA6 in human cytotrophoblast invasion and migration were studied. MATERIALS AND METHODS: Human placenta tissues were collected from normal pregnancies and pregnancies complicated by early-onset severe preeclampsia (sPE). Immunofluorescence, RT-qPCR and Western blotting were used to determine NCOA6 expression. Transwell invasion/migration assays were performed to explore whether NCOA6 knockdown affected human placenta-derived HTR-8/SVneo cell invasion/migration. Gelatin zymography was performed to examine the change in the gelatinolytic activities of secreted MMP2 and MMP9. Luciferase reporter assays were used to explore whether NCOA6 coactivated NF-κB-mediated MMP9 transcription. RESULTS: NCOA6 is mainly expressed in the human placental trophoblast column, as well as in the EVTs. HTR-8/SVneo cell invasion and migration were significantly attenuated after NCOA6 knockdown, and the secretion of MMP9 was decreased due to transcriptional suppression. NCOA6 was further found to coactivate NF-κB-mediated MMP9 transcription. Moreover, expression of NCOA6 was impaired in placentas of patients complicated by early-onset sPE. CONCLUSIONS: Thus, we demonstrated that NCOA6 is important for cytotrophoblast invasion/migration, at least partially, by activating NF-κB-mediated MMP9 transcription; the downregulation of NCOA6 may contribute to the pathogenesis of early-onset sPE.

Placensin is a glucogenic hormone secreted by human placenta.

FBN1 encodes asprosin, a glucogenic hormone, following furin cleavage of the C-terminus of profibrillin 1. Based on evolutionary conservation between FBN1 and FBN2, together with conserved furin cleavage sites, we identified a peptide hormone placensin encoded by FBN2 based on its high expression in trophoblasts of human placenta. In primary and immortalized murine hepatocytes, placensin stimulates cAMP production, protein kinase A (PKA) activity, and glucose secretion, accompanied by increased expression of gluconeogenesis enzymes. In situ perfusion of liver and in vivo injection with placensin also stimulate glucose secretion. Placensin is secreted by immortalized human trophoblastic HTR-8/SVneo cells, whereas placensin treatment stimulates cAMP-PKA signaling in these cells, accompanied by increases in MMP9 transcripts and activities, thereby promoting cell invasion. In pregnant women, levels of serum placensin increase in a stage-dependent manner. During third trimester, serum placensin levels of patients with gestational diabetes mellitus are increased to a bigger extent compared to healthy pregnant women. Thus, placensin represents a placenta-derived hormone, capable of stimulating glucose secretion and trophoblast invasion.

Factors predicting clinical pregnancy rate of in vitro fertilization-embryo transfer (a STROBE-compliant article).

The aim of this study was to investigate the factors predicting clinical pregnancy rate of in vitro fertilization-embryo transfer (IVF-ET).The data of 9960 patients receiving IVF-ET fresh cycle at our Reproductive Center from January 2009 to December 2017 were first divided into pregnant group and non-pregnant group to find the clinical pregnancy rate-related factors. According to the serum HCG levels at 36 hours and 12 hours after HCG trigger, all patients were divided into 4 groups including <50 mIU/ml, ≥50 and <100 mIU/ml, ≥100 and <200 mIU/ml, and ≥200 mIU/ml groups to know whether the HCG levels at 36 hours and 12 hours affect the pregnancy rate. According to the serum HCG ratio at 36 hours to 12 hours (36 h/12 h) after HCG trigger, all patients were divided into three groups including <0.88, 0.88-1.06 and >1.06 groups to observe whether the serum HCG ratio (36 h/12 h) affects the clinical pregnancy rate. According to different assisted pregnancy modes, all patients were divided into 3 groups including IVF, ICSI, and IVF/ICSI groups to observe whether the assisted pregnancy mode affects the clinical pregnancy rate. The correlation of the clinical pregnancy rate with pregnancy rate-related factors obtained above was analyzed using logistic regression analysis model.The clinical pregnancy rate significantly increased (P < .01) in the HCG ratio (36 h/12 h) >1.06 group as compared with the HCG ratio (36 h/12 h) < 0.88 and 0.88-1.06 groups. The serum estrogen (E2) level at 36 hours was significantly lower and the number of retrieved oocytes was significantly higher in the HCG ratio (36 h/12 h) >1.06 group than in the HCG ratio (36 h/12 h) <0.88 and 0.88-1.06 groups (P = .000).The serum HCG ratio (36 h/12 h) may be used as a predictor of IVF-ET clinical pregnancy rate. High clinical pregnancy rate is probably associated with E2 down-regulation in the HCG ratio (36 h/12 h) >1.06 group.

Impact of Female Obesity on Cumulative Live Birth Rates in the First Complete Ovarian Stimulation Cycle.

Background: Female overweight/obesity has been reported to be associated with compromised pregnancy outcomes in fresh embryo transfer cycles. It is unclear whether the cumulative live birth rate (CLBR) is adversely affected after all viable embryos are transferred from the first ovarian stimulation cycle. Objectives: To investigate whether the CLBR was compromised in obese women. Method: A total of 9,772 young women underwent their first IVF/ICSI cycles from January 2012 to October 2017. Pregnancy outcomes were compared according to female BMI. Results: Among 1,671 women with polycystic ovary syndrome (PCOS), those with a BMI ≥ 28 kg/m2 had a lower cumulative clinical pregnancy rate (CCPR) and CLBR during the first complete ovarian stimulation cycle. Additionally, the pregnancy loss rate was increased in this group, although the difference was not significant. Among the 8,101 women without PCOS, the CCPR and CLBR of obese patients was also significantly decreased, and this group also showed increased pregnancy loss rates. Moreover, overweight women also had a decreased CLBR. Conclusions: Female obesity adversely affected the CLBR after utilizing the viable embryos from first oocytes retrieval.

miR-181a-5p suppresses invasion and migration of HTR-8/SVneo cells by directly targeting IGF2BP2.

Pre-eclampsia is a pregnancy-related disease that may cause maternal, neonatal and fetal morbidity and mortality and exists in 3-5% of pregnancies worldwide. The discovery of dysregulated microRNAs and their roles in placental development has provided a new avenue for elucidating the mechanism involved in this pregnancy-specific disorder. Here, the roles of human miR-181a-5p, a microRNA that is increased in both the plasma and placenta of severe pre-eclamptic patients, in invasion and migration of trophoblasts were investigated. Ectopic-expression of miR-181a-5p impaired the invasion and migration of HTR-8/SVneo cells, whereas miR-181a-5p inhibition had the opposite effects. IGF2BP2, which harbors a highly conserved miR-181a-5p-binding site within its 3'-UTR, was identified to be directly inhibited by miR-181a-5p. Moreover, siRNAs targeting IGF2BP2 imitated the effects of overexpressed miR-181a-5p on HTR-8/SVneo cell invasion and migration, whereas restoring IGF2BP2 expression by overexpressing a plasmid encoding IGF2BP2 partially reversed the studied inhibitory functions of miR-181a-5p. Thus, we demonstrated here that miR-181a-5p suppresses the invasion and migration of cytotrophoblasts, and its inhibitory effects were at least partially mediated by the suppression of IGF2BP2 expression, thus shedding new light on the roles of miR-181a-5p in the pathogenesis of severe pre-eclampsia.

Influence of endometrial thickness on treatment outcomes following in vitro fertilization/intracytoplasmic sperm injection.

BACKGROUND: The study was designed to investigate the roles of endometrial thickness (EMT) at the day of human chorionic gonadotropin (hCG) administration on pregnancy outcomes in a large patient population. METHODS: This retrospective cohort study included 9,952 patients undergoing their first IVF/ICSI with autologous oocytes from January 2011 to January 2015. Patients were divided into three groups based on the EMT (group A:≤8 mm; group B: 9-14 mm and group C:≥15 mm). Live birth rate (LBR), clinical pregnancy rate (CPR), early miscarriage rate (EMR), and ectopic pregnancy rate (EPR) were analyzed. Additionally, the live birth rate was analyzed for patients with single or double gestational sacs. RESULTS: Significant differences (p < 0.05) were detected in the LBRs (30.38%, 45.73% and 54.55% for groups A, B, and C, respectively), CPRs (38.57%, 55.04% and 64.32%, respectively), and EPRs (5.58%, 3.48% and 2.19%, respectively), with thicker endometrial thickness favoring all three parameters. However, no differences were found in the EMRs among the three groups (15.64%, 13.44% and 13.05%, respectively, p > 0.05). After adjusting for female age, body mass index (BMI) and endometrial pattern, the multivariate logistic regression analysis demonstrated that the associations between EMT and LBR (adjusted OR: 2.645; 95% CI 2.020-3.464; p < 0.01), CPR (adjusted OR 2.693 95% CI 2.012-3.605 p < 0.01), and EPR (adjusted OR: 0.298 95% CI 0.101-0.713; p < 0.05) were significant. Additionally, live birth rates in the double gestational sac group were different (p < 0.05) among patients with different EMT (72.73%, 87.28%, and 87.36%, respectively), whereas no difference was found in the single gestational sac group. In the double gestational sac group, LBR was positively correlated with increasing endometrial thickness only in patients with twin pregnancies but not in patients with singletons. CONCLUSIONS: Our study shows that endometrial thickness at the day of hCG administration has an effect on LBR, CPR and EPR, with all three parameters increasing with the EMT. Furthermore, successful twin pregnancies are associated with a thicker endometrium.

[Effects of Ca2+ on nitric oxide-induced adventitious rooting in cucumber under drought stress]. / å¹²æ—±æ¡ä»¶ä¸‹é’™ç¦»å­å¯¹ä¸€æ°§åŒ–æ°®è¯±å¯¼é»„ç“œä¸å®šæ ¹å‘ç”Ÿçš„å½±å“.

Cucumber (Cucumis sativus L. 'Xinchun 4') was used to explore the relationship between nitric oxide (NO) and calcium (Ca2+) during adventitious rooting under drought stress. Rooting parameters, endogenous Ca2+ fluorescent intensity and the antioxidant enzymes activity (SOD, CAT and APX) in cucumber explants under drought stress were investigated. The results showed that treatment with 200 µmol·L-1 CaCl2 and 0.05% PEG significantly improved the number and length of adventitious root in cucumber explants under drought stress, while the application of Ca2+ chelating agent (EGTA) and channel inhibitor (BAPTA/AM) significantly decreased NO-induced number and length of adventitious root under drought stress. Under drought stress, the fluorescence intensity of Ca2+ in hypocotyls treated with NO and CaCl2 was improved, however, the Ca2+ fluorescence intensity in the hypocotyls treated with NO scavenger (cPTIO) was significantly lower than that in the hypocotyls treated with NO. Under drought stress, the activities of antioxidant enzymes in the cucumber explants were significantly promoted by the treatments with NO and CaCl2, however, Ca2+ chelating agent and channel inhibitor significantly decreased the activity of antioxidant enzymes induced by NO. In conclusion, Ca2+ might be involved in the process of NO-adjusted antioxidant enzymes activity during adventitious rooting under drought stress, which alleviated the negative effects of drought on the adventitious rooting and promoted the formation of adventitious roots.

[Effects of exogenous salicylic acid and brassinolide on photosynthesis of cucumber seedlings under low temperature stress.] / æ°´æ¨é ¸å’Œæ²¹èœç´ å† é ¯å¯¹ä½Žæ¸©èƒè¿«ä¸‹é»„ç“œå¹¼è‹—å ‰åˆä½œç”¨çš„å½±å“.

An experiment was conducted to investigate the mechanism of the regulation of salicylic acid (SA) and 2,4-epibrassionolide (EBR) on photosynthesis in cucumber (Cucumis sativa) seedlings under low temperature stress. Taking cucumber cultivar 'Youbo1-5' as material, the seedlings were pre-treated with 1 mmol·L-1 SA or 0.1 µmol·L-1EBR (sprayed once a day), and then were exposed to chilling temperature (10 ℃/5 ℃, PFD 80 µmol·m-2·s-1) after being pre-treated 2 days. The results showed that the growth and net photosynthetic rate (Pn) of cucumber seedlings were decreased under low temperature stress. However, the Pn, maximal photochemical efficiency of PS2 (Fv/Fm), actual photochemical efficiency of PS2 (ΦPS2) and photochemical quenching coefficient (qP) were significantly improved in SA- and EBR-pretreated seedlings, and the increase range of non-photochemical quenching coefficient (NPQ) was decreased. Moreover, the activities of ribulose 1,5-biphosphate carboxylase/oxygenase (Rubisco), sedoheptulose-1,7-bisphosphatase (SBPase), fructose-1,6-bisphosphate aldolase (FBA) and transketolase (TK) were signi-ficantly increased. These findings suggested that SA and EBR improved photosynthesis of cucumber seedlings by promoting the activities of key enzymes and increased low temperature tolerance.

Molecular karyotype single nucleotide polymorphism analysis of early fetal demise.

We explored the application of single nucleotide polymorphism microarray (SNP array) in molecular karyotype analysis for early spontaneous abortion detection in assisted reproductive technology (ART). SNP array was performed in 81 cases. Of the 81 cases, 16 experienced natural conception (NC) and 65 were pregnant by ART. Of the 65 cases, 4 underwent artificial insemination (AI), 32 fresh in vitro fertilization-embryo transfer (IVF-ET), 9 fresh intracytoplasmic sperm injection (ICSI), and 20 thawed embryo transfer. In the 81 cases examined 69.1% displayed an abnormal molecular karyotype. In the subjects greater than 35 years of age, the abnormal molecular karyotype rate was 87.5% higher compared to 61.4% in younger individuals (P < 0.05). There was no significant difference in the abnormal molecular karyotype rate or type between ART (64.6%) and NC (87.5%). Compared with traditional cytogenetic diagnosis, the SNP array can identify a greater number of abnormal karyotypes.

[MicroRNA expression and its role in the cell cycle regulation in decidualized endometrial stromal cells in vitro].

OBJECTIVE: To study microRNA (miRNA) expression and role of cell cycle regulation in decidualized endometrial stormal cells (ESC) in vitro. METHODS: ESC was induced decasualization in vitro and matched with non-decidualized cells as controls. The expression repertoire of miRNA was measured by microarray chip and was validated by real-time PCR. Flow cytometry was used to identify ESC cycle during decidual reaction in vitro and after miRNA222 inhibitor was transfected into it. RESULTS: (1) Between decidualized and undecidualized stromal cells, there were 49 miRNAs significantly different expression by microarray chip, including 16 miRNA up-regulation and 33 miRNA down-regulation.hsa-miR-27b, 30c, 143, 101, 181b, 29b, 30d, 507, 23a, 222, 221 exhibited significantly differential expression between decicualized and undecidualized stromal cells by real-time PCR (P < 0.05). (2) After miRNA222 inhibitor (NC-FAM) transfection to decidual ESC, ESC were cultured by FBS medium for 24 hours, the rate of transfection was 70%. ESC were transfected with miRNA 222 inhibitor and cultured for 48 hours, the percentage of ESC at S-phase of (6.2 ± 0.7)% were significantly lower than (10.9 ± 0.8)% in control group (P < 0.05);the percentage of ESC at G(0)/G(1) phase increased at transfection group [(77.5 ± 1.3)% vs. (73.0 ± 1.6)% at control group], but there was no significant difference (P > 0.05). Decasualization ESC were transfected with miRNA 222 inhibitor and cultured for 48 h, the percentage of ESC at S-phase was (3.3 ± 0.6)% in transfection group, which were significantly lower than (7.8 ± 0.9)% in control group (P < 0.05). The percentage of ESC at G(0)/G(1) phase was (80.7 ± 1.6)% in transfection group and (74.9 ± 1.1)%. In control group, which did not reached statistical difference (P > 0.05). CONCLUSION: miRNA was involved in ESC decidual process in vitro by regulating cell cycle.

Relationship between processed total motile sperm count of husband or donor semen and pregnancy outcome following intrauterine insemination.

We retrospectively analyzed 6,360 artificial insemination cycles of husband's semen through intrauterine insemination (AIH-IUI) or artificial insemination with donor semen through intrauterine insemination (AID-IUI) in patients with infertility between August, 1998 and August, 2010. The relationship between processed total motile sperm count (PTMS) and pregnancy outcome was determined. The study was divided into 6 groups according to PTMS. Group 1: ≤ 2.0 million, Group 2: 2.1-4.0 million, Group 3: 4.1-6.0 million, Group 4: 6.1-8.0 million, Group 5: 8.1-10.0 million, and Group 6: >10.0 million. There was no statistically significant difference in age, duration of infertility, unilateral tubal patency, induced ovulation, and single IUI or double IUI between the 6 groups in both AIH-IUI and AID-IUI. The total clinical pregnancy rate of AIH-IUI was 10.81 % and AID-IUI was 27.52 %. Among the 6 groups, the clinical pregnancy rate was the lowest in Group 1 (P < 0.05) in both AIH-IUI and AID-IUI. With the increased PTMS, the clinical pregnancy rate of IUI was improved. However, a statistical difference between groups was only observed for Group 1. When PTMS is ≤ 2 × 10(6) the clinical pregnancy rate of IUI is significantly decreased. In this case in vitro fertilization (IVF) should be adopted.

Percutaneous nephrolithotomy through the upper pole calix access for complicated renal calculi: report of 581 cases / 南方医科大学学报

<p><b>OBJECTIVE</b>To discuss the optimal approach of percutaneous nephrolithotomy (PCNL) for treatment of complicated renal calculi.</p><p><b>METHODS</b>A total of 581 patients with complicated renal calculus were treated by PCNL through the upper pole calix access. Of the 581 patients, 55 had multiple upper pole calculi, 136 had staghorn stones, 145 had partial staghorn stones, and 245 had multiple renal calculi.</p><p><b>RESULTS</b>PCNL through the upper pole calix access was completed successfully in all the cases. Of these patients, 90.3% (525/581) were stone-free after a single access, with a total stone-free rate of 94.6% (550/581). Thirty-five patients needed two accesses, 10 needed 3 accesses, 2 required 4 accesses, and 1 patients had 5 accesses. The operative time ranged from 30 to 150 min (mean 45 min). The successful rate of puncture was 100% without occurrence of severe injury of the pleura, intestine, peritoneum or other adjacent organs.</p><p><b>CONCLUSIONS</b>Percutaneous nephrolithotomy through the upper pole calix access allows greater stone clearance rate due to its easy access into the intrarenal collecting system and can be an ideal approach for PCNL for complicated renal calculi.</p>

Laparoendoscopic single-site surgery for male pseudohermaphroditism: the initial report / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the therapeutic effect of laparoendoscopic single-site surgery (LESS) for treatment of male pseudohermaphroditism.</p><p><b>METHODS</b>A 17-year-old patient with male pseudohermaphroditism and a female social sex was admitted. According to the request by the patient and the relatives for a female gender, LESS vaginoplasty and cryptorchidectomy were performed using a single multilumen port inserted through a 2.5 cm incision below the umbilicus, followed by reconstruction of the perineal region by open surgery.</p><p><b>RESULTS</b>The total operative time was 7 h, and the LESS procedure lasted for about 3.5 h. No other port incision was needed. The estimated intraoperative blood loss was 400 ml. No electrolyte or metabolic acid-base balance disorders were observed perioperatively. In the follow-up examination at 6 months after the operation, the reconstructed vagina healed smoothly without obvious contraction or fixation failure, and the perineal region showed good appearance.</p><p><b>CONCLUSION</b>With minimal invasiveness, LESS surgery produces good cosmetic effect and allows rapid postoperative recovery, thus may become a promising alternative to the management of pseudohermaphroditism.</p>

Laparoendoscopic single-site surgery radical cystectomy with orthotopic taenia myectomy sigmoid neobladder: initial report / 南方医科大学学报

<p><b>OBJECTIVE</b>To report the first case and detailed techniques of laparoendoscopic single-site surgery (LESS) radical cystectomy with orthotopic taenia myectomy sigmoid neobladder for organ-confined bladder cancer.</p><p><b>METHODS</b>A 74-year-old man presented with gross hematuria for 2 months and biopsy revealed bladder cancer. LESS radical cystectomy and bilateral pelvic lymphadenectomies were performed using a single multilumen port inserted through a solitary 3.5 cm lower abdominal incision with conventional laparoscopic instruments. The taenia myectomy sigmoid pouch was then constructed by open procedure.</p><p><b>RESULTS</b>The total operative time was 9.5 h, and the LESS procedure lasted for about 5.5 h. No other port incision was added. The final pathology revealed urothelial carcinoma. The estimated intraoperative blood loss was 600 ml with blood transfusion of 400 ml. The pelvic lymph nodes and the surgical margins of the ureters and urethra were all free of tumor invasion. No water electrolyte and metabolic acid-base balance disorders were observed perioperatively. The neobladder capacity was about 280 ml, with a residual urine volume of 10 ml and peak flow rate of 11.1 ml/s 3 months postoperatively.</p><p><b>CONCLUSION</b>Although with a steep learning curve, LESS surgery can be a less invasive and promising alternative to muscle-invasive bladder carcinoma.</p>

Laparoscopic radical cystectomy and detenial sigmoid colon orthotopic neobladder reconstruction for bladder tumor in a 3-year-old boy / 南方医科大学学报

<p><b>OBJECTIVE</b>To present a case of laparoscopic radical cystectomy and detenial sigmoid colon orthotopic neobladder reconstruction for bladder tumor in a child.</p><p><b>METHODS</b>A 3-year-old boy with bladder rhabdomyosarcoma underwent laparoscopic radical cystectomy and detenial sigmoid colon orthotopic neobladder reconstruction. The bilateral pelvic lymphadenectomy and cystectomy were performed laparoscopically, and removal of the mobilized specimens and urinary diversion operation were managed through enlarged abdomen incision. The urinary diversion procedure included construction of the detenial sigmoid pouch, bilateral stented antiflux implantation of the ureters in the pouch and orthotopic anastomosis of the neobladder with the urethra.</p><p><b>RESULTS</b>The total operative time was 6 h, and the laparoscopic procedure lasted for about 3.5 h. The intraoperative blood loss was 50 ml, and 200 ml concentrated red blood cell transfusion was used for the safety of the patient. Six dissected lymph nodes in each pelvic side and the surgical margins of the ureter and urethra were all free of tumor invasion. Bowel peristalsis recovered 3 days after the operation, and the pelvic drainage and the neobladder drainage tubes were removed on day 7 and 14, respectively. The urethral catheter and ureteral stents were removed 25 days after the operation. The daytime urine control and micturition recovered 1 week after the operation. The neobladder capacity was about 110 ml, with residual urine volume of 10 ml and peak flow rate of 12 ml/s after 5 months. No perioperative complications occurred such as water-electrolyte and metabolic acid-base balance disorders, urinary leakage, reflux or bowel obstruction.</p><p><b>CONCLUSION</b>Laparoscopic radical cystectomy is minimally invasive, reduces intraoperative blood loss and allows rapid postoperative recovery, and can be a promising approach to management of bladder rhabdomyosarcoma in children.</p>

Reversion transcriptional expression of DAPK in bladder cancer T24 cells 5-aza-2'-deoxycytidine / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the methylation status of the promoter of resion death associated protein kinase (DAPK) gene in bladder cancer cell (T24), and study the effect of 5-aza-2'-deoxycytidine (5-aza-dc) on DAPK gene reactive expression in T24 and its inhibitory effect on T24.</p><p><b>METHODS</b>The bladder cancer cell T24 was treated with different doses of 5-aza-dc. The inhibitory effect and apoptosis rate were detected by MTT and flow cytometry, and the changes of DAPK mRNA and protein expression and the methylation status of DAPK promoter were assessed by RT-PCR, Western blotting, and methylation specific PCR, respectively.</p><p><b>RESULTS</b>The growth of bladder cancer cell was inhibited significantly and the maximal apoptosis rate detected by flow cytometry was (24.12-/+1.4)%. DAPK mRNA was not expressed in bladder cancer cell T24 in normal conditions. DAPK mRNA and protein re-expressed after 5-aza-dc (12.5 micromol/L) treatment in cell line T24 for 24 h, and DAPK promoter became unmethylated.</p><p><b>CONCLUSIONS</b>The promoter methylation can be an important factor for silencing the expression of DAPK in bladder cancer cell. 5-aza-dc can inhibit the growth and induce apoptosis of bladder cancer cells through reversing unmethylation status of DAPK promoter.</p>

Preparation of whole-kidney acellular matrix in rats by perfusion / 南方医科大学学报

<p><b>OBJECTIVE</b>To prepare rat whole-kidney acellular matrix (ACM) scaffolds using fluid perfusion method.</p><p><b>METHODS</b>The kidneys with ureters and renal vessels were harvested from 12-week-old Wistar rats. Intravenous catheters were inserted through the renal arteries to establish channels for whole-kidney retrograde perfusion successively with heparinized PBS, 1% SDS, deionized water, 1% TritonX-100 and antibiotic-containing PBS under a pressure of 100 cmH2O. After decellularization, the scaffolds were observed under microscope with HE staining, scanning electron microscope, and fluorescence microscope with DAPI fluorescence staining.</p><p><b>RESULTS</b>No cell residue was found in the scaffolds under microscope. Scanning electron microscope identified reticular structures consisting of basilar membrane and collagen without normal cellular structures in the scaffolds, and no strong fluorescence due to the binding of DAPI to the cell nuclei was observed under fluorescence microscope.</p><p><b>CONCLUSION</b>Fluid perfusion is simple and reliable to prepare rat whole-kidney acellular matrix, which may serve as an ideal cell-free scaffold.</p>

Environment-friendly determination of low concentration azobenzene beta-cyclodextrin-modified electrode / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To study environment-friendly determination of azobenzene in trace amounts using beta-cyclodextrin (beta-CD)-modified Au electrode.</p><p><b>METHODS</b>beta-CD-modified Au electrode was fabricated with a two-step approach, and then a gold electrode modified with beta-CD was used to detect azobenzene by employing Osteryoung square wave voltammetry.</p><p><b>RESULTS</b>The modified electrode could detect azobenzene, showing a good linearity between the electrochemical current and concentration.</p><p><b>CONCLUSION</b>Although the electrochemical current is related with concentration, the detection limit is around 1.0 x 10(-10) mol/L. This study may provide a new environment-friendly approach for monitoring water quality.</p>

Cytocompatibility of silk fibroin film with rabbit urinary bladder transitional epithelial cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To obtain large quantities of well differentiated urinary bladder transitional epithelial cells for used as the seed cells in bladder tissue engineering, and evaluate the cytocompatibility of silk fibroin film with the transitional cells in vitro to assess the possibility of tissue-engineered urinary organ construction.</p><p><b>METHODS</b>The urinary bladder transitional epithelial cells were isolated from the bladders of New Zealand rabbits and cultured in vitro as the seed cells, whose morphology was observed and the specific protein (cytokeratin) expression identified by immunofluorescence assay. The cells were seeded in 96-well plates at 1 x 10(>4)/ml and incubated with silk fibroin film leaching solution or culture medium (negative control). MTT assay was performed to determine the cell proliferation rates of the wells and evaluate the cytotoxicity and cytocompatibility of the silk fibroin film.</p><p><b>RESULTS</b>The isolated urinary bladder transitional epithelial cells reached confluence after 9-10 days of culture, which showed positive staining for immunocytochemistry with monoclonal antibody against cytokeratin. The absorbance of the cells culture in the presence of silk fibroin film leaching solution averaged 0.424-/+0.020, 0.996-/+0.118 and 1.285-/+0.048 after at 24, 72 and 120 h of cell culture, and that of the negative control group at the time points was 0.419-/+0.030, 1.105-/+0.098 and 1.228-/+0.052, respectively, showing no significant difference between the two groups (P>0.05).</p><p><b>CONCLUSION</b>Silk fibroin film has good cytocompatibility with rabbit urinary bladder transitional epithelial cells, and may serve as good scaffold material for urologic tissue engineering.</p>