Objective To investigate factors influencing islet isolation and purification in dogs and to explore how to obtain massive purified viable islets from canine pancreas.To provide experimental evidence for pancreatic islet transplantation in clinical practice.Methods Islets were isolated from pancreas of 22 dogs by modified automated techniques,followed by purification using continuous density gradients.The purity of islets was determined by dithizone (DTZ) staining,and the function of islets was evaluated by glucose stimulating insulin release test.Results The crude islets yield was ( 155 040 ±310) IEQ/pancreas while the yield was (74 200 ±185) IEQ/pancreas after purification.The average purity was (89.4 ±2.6) ％.The recovery rate was(47.8 ±1.3) ％.Islet viability was(93.0 ± 1.7)％.The modified techniques improved both the yield and the quality in islet isolation and purification.The islets were morphologically intact after purification.Glucose stimulating insulin release test showed that the difference of the secreted insulin concentration had statistical significance ( P ＜0.001 ) between low concentration glucose group and high concentration glucose group.Conclusion It is of great value for clinical research to obtain enough islets through islet isolation and purification in dogs.

Objective To determine any relationship among detrusor wall thickness (DWT),bladder capacity (BC) and neurogenic lower urinary tract dysfunction (NLUTD),and to study the clinical implications of using DWT to predict the risk of renal injury after spinal cord injury (SCI).Methods Forty-eight adults with NLUTD due to SCI were recruited for the SCI group,and another 41 healthy adults formed a control group.Both groups underwent urodynamic evaluation and maximum bladder capacity (MBC) detection.The DWT of all subjects was measured by ultrasound imaging of the anterior bladder wall.Thereafter,the SCI group was divided into low and high risk subgroups based on the results of the urodynamic tests.Patients with detrusor leak point pressure ＜40 cmH2O were classified as low risk.Results At MBC,the average DWT in the SCI group was (0.97 ± 0.31)mm and (0.59 ± 0.08)mm in the control group,demonstrating a significant difference in DWT between patients with NLUTD and normal adults.The average DWT among the patients with type A detrusor sphincter dyssynergia was (1.10 ± 0.34)mm,which was significantly higher than among the patients with external urethral sphincter contraction without detrusor contraction (type C).DWT was significantly correlated with detrusor leak point pressure.A DWT of 0.87 mm (sensitivity 89.5％,specificity 58.6％) could be used as a critical point for predicting risk of renal injury in patients with NLUTD.Conclusions DWT is significantly greater among SCI patients with NLUTD and it correlates positively with detrusor leak point pressure.DWT could be used as a risk predictor for renal injury in patients with NLUTD due to SCI.

Objective To analyse the breast conserving surgery feasibility in patients with advanced breast cancer after neoadjuvant chemotherapy.Methods Sixty patients with advanced breast cancer were collect-ed from June 2010 to June 2011, and were divided into breast conserving surgery after neoadjuvant chemotherapy group and modified radical mastectomy group according to the intention,30 cases in each group.They were given breast conserving surgery after neoadjuvant chemotherapy and modified radical mastectomy respectively,with fol-lowing-up for 3 years.The treatment effect and contrast analysis of two groups of breast cancer CTCs positive of the two groups were compared,two groups of late follow-up of patients with local recurrence and distant metasta-sis rate,overall survival and disease free survival rate were recorded.Results Compared with the modified radi-cal mastectomy group,CTCs detection rate in breast conserving surgery group had no statistical differences(P>0.05).Two groups of patients in clinical complete remission rate,partial remission rate,disease stability factor and local recurrence and distant metastasis rate,overall survival and disease free survival rate had no statistical significances(P>0.05).Conclusion The application of breast conserving surgery after neoadjuvant chemother-apy for patients with advanced breast cancer can achieve similar effect as modified radical mastectomy surgery treatment,and can be used as a effective treatment for advanced breast cancer.

Objective To evaluate one-stage procedure for the treatment of neonatal intermediate and high imperforated anus. Methods Clinical data of 21 neonates with intermediate or high imperforated anus undergoing one-stage surgical procedure were summarized. Results Postoperative complication included mucous overlap and soiling in 1 case, and occasional soiling in 2 cases. No postoperative constipation developed in any case. Conclusions Neonatal one-stage procedure avoids short comings often seen in staged procedures and needs no colostomy, promoting hyperplasia and prolongs sphincter during early stage, preventing secondary megacolon.

Objective To investigate the effectiveness of botulinum toxin A (BTX-A) injection to the external urethral sphincter guided by ultrasound in the treatment of various types of lower urinary tract dysfunction.Methods Twenty-two patients with urinary retention caused by neurological dysfunction (neurogenic bladder) were recruited,of which 18 were men and 4 women.They had various types of lower urinary tract dysfunction including detrusor external sphincter dyssynergia (DESD) in 7,nonrelaxing urethral sphincters in 5,and detrusor underactivity in 10.BTX-A (100 U) was injected into the external urethral sphincter at three different points guided by ultrasound.Clinical effects and urodynamic parameters were compared at baseline and after treatment.Results After treatment the urinary function and urodynamic parameters in each group improved significantly compared with the baseline.Maximum efficacy was observed at the 4th week.In the DESD group,3 patients (43％) had an excellent result and 3 (43％) had significant improvement; in the nonrelaxing sphincter group 3 (60％) had excellent results and 2 (40％) showed significant improvement; in the detrusor underactivity group 5 (50％) had excellent results and 3 (30％) had significant improvement.There were no obvious side effects.Conclusions BTX-A injections guided by ultrasound can accurately locate the external urethral sphincter.It is a simple and repeatable method for the treatment of lower urinary tract dysfunction.

OBJECTIVETo investigate the effects of fresh and preserved amniotic membrane on polymorphonuclear neutrophils (PMNs) so as to understand the anti-inflammatory mechanism of amniotic membrane transplantation.

METHODSConditioned medium was collected 48 hours after fresh or preserved amnions were cultured in DMEM and 5% CO(2) at 37 degrees C. Then, polymorphonuclear cells were cultured in conditioned culture or DMEM. Fluorescent microscopy with 4',6-diamidino-2-phenylindole (DAPI) staining and cytometry were performed 6, 9, 12, and 15 hours later.

RESULTSApoptotic neutrophils were found in each group at different time points. The percentage of apoptotic cells at 6, 9, 12, and 15 hours after culture in the fresh and preserved amnion groups and the control group was 17.3%, 24.4%, 29.8%, 37.1%, and 16.2%, 20.1%, 23.7%, 27.7%, and 10.2%, 13.7%, 21.1%, 26.4%, respectively (t test, P(1) < 0.01, P(2) < 0.01 and P(3) < 0.01).

CONCLUSIONAmniotic membrane can accelerate apoptosis of polymorphonuclear neutrophils, reduce inflammation, and prevent ocular surface collagen from resolution, indicating that fresh amnion might have a stronger effect than preserved amnion.

The cloned cDNA sequence of rice (Oryza sativa L. Cpslo17) chitinase gene Oschi was cloned, (which was registered in GenBank, the accession number: EU045451) ligated with the expression vector pGEX-4T-1, and transformed into E. coli BL21(DE3). The expression of Oschi was induced by IPTG, and the conditions were optimized. After purification the in vitro activity of Oschi chitinase was analyzed, and the results indicated that it could efficiently degrade chitin.
Chitin ; metabolism ; Chitinases ; biosynthesis ; genetics ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Molecular Sequence Data ; Oryza ; enzymology ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; metabolism

Objective:To explore the efficacy and safety of TiRobot combined with three-dimensional imaging in the minimally invasive surgery for pelvic fractures.Methods:A retrospective analysis was conducted of the 40 patients with pelvic fracture who had been treated by fixation with S1 and S2 sacroiliac screws at Department of Orthopaedics and Traumatology, Yangjiang People's Hospital from January 2019 to May 2021. They were divided into 2 groups according to their treatment methods. In the TiRobot group of 20 cases subjected to percutaneous sacroiliac screw fixation assisted by TiRobot combined with three-dimensional imaging, there were 13 males and 7 females with an age of (38.2 ± 8.8) years. In the manual group of 20 cases subjected to fixation with manual placement of sacroiliac screws under conventional C-arm fluoroscopy, there were 11 males and 9 females with an age of (37.3 ± 9.2) years. The 2 groups were compared in terms of fluoroscopy time for screw placement, guide needle adjustment, operation time, intraoperative blood loss, visual analogue scale (VAS) 72 hours after operation, postoperative hospital stay, time to ambulation, excellent to good rate of screw placement, complication rate, fracture union time, Majeed score at 6 months after operation, and excellent to good rate of functional evaluation.Results:There was no significant difference between the 2 groups in their preoperative general data, showing they were comparable ( P > 0.05). In the TiRobot group, fluoroscopy time for screw placement [(8.2 ± 2.9) s], guide needle adjustment [(0.4 ± 0.2) times], operation time [(67.4 ± 5.5) min], and intraoperative blood loss [(36.5 ± 8.0) mL] were significantly less than those in the manual group [(40.4 ± 4.5) s, (8.6 ± 0.7) times, (78.4 ± 7.2) min, and (41.6 ± 7.8) mL], postoperative VAS [3.0 (4.0, 5.0) points] was significantly lower than that in the manual group [4.0 (5.0, 6.0) points], the excellent to good rate of screw placement (100%, 40/40) was significantly higher than that in the manual group (85.0%, 34/40), and the complication rate (5.0%,1/20) was significantly lower than that in the manual group (35.0%, 7/20) (all P < 0.05). There was no significant difference between the 2 groups in postoperative hospital stay, time to ambulation, fracture union time, Majeed score, or excellent to good rate of functional evaluation ( P > 0.05). Conclusion:In the minimally invasive surgery for pelvic fractures, TiRobot combined with three-dimensional imaging leads to positive outcomes, because it can reduce operation time and radiation exposure, improve accuracy of screw placement, and increase safety.

Objective To analyze the differential protein expression of three negative breast cancer tissue before and after menopause application of iTRAQ technology, to explore the effects of menopause on three negative breast cancer. Methods Selected premenopausal and postmenopausal women with 8 cases in each by negative pathological diagnosis, differential protein analysis on significant function, difference of protein pathway and difference of protein verify were analyzed by the application iTRAQ technology. Results (1﹚Differential protein interactome was relatively centralized premenopausal, and postmenopausal differences of protein was much, and the distribution of the dispersed. (2﹚In pre menopausal cancer tissue protein differences existed in 5 of the significant difference in differences approach; post-menopausal cancer tissue protein existsed in 15 significantly different pathways. (3﹚Tumor adjacent tissues compared with premenopausal had a total of 214 significant difference proteins, postmenopausal had a total of 360 significant differ-ence proteins. The upregul ated proteins in 81 kinds of premenopausal three negative breast cancer tissues, 133 down regμl ated proteins, postmenopausal up-regμl ated protein 157 types, down 203. Conclusion Using iTRAQ technology to found that the expression has a certain particμl arity differences in three negative breast cancer before and after menopause, indicate three negative breast cancer may have different pathogenesis in the different estrogen environment, may be a new target for treatment of TNBC.