1.Laparoscopic surgery for high-risk prostate cancer:urinary and oncologic outcomes of vesicourethral anastomosis with maximal urethral length and bladder neck preservation
Kun ZHENG ; Xiaoyong HU ; Qiang FU ; Wang LI ; Ying WANG ; Nailong CAO ; Jiasheng CHEN ; Ranxing YANG
Journal of Modern Urology 2024;29(7):612-616
Objective To explore the application value of vesicourethral anastomosis with maximal urethral length preservation(MULP)and bladder neck preservation(BNP)in laparoscopic radical prostatectomy(LRP)or robot-assisted laparoscopic radical prostatectomy(RALP)for high-risk prostate cancer(HRPC)in terms of early urinary continence and oncology.Methods Clinical data of 23 HRPC patients who underwent LRP(including RALP)with MULP and BNP in our hospital during May 2022 and Jan.2024 were retrospectively analyzed.Patients'basic information,surgical parameters,postoperative complications,oncological outcomes and urinary incontinence were collected and analyzed.Results All operations were completed successfully without conversion to open surgery.The operation time was(108±31)min,average blood loss(112±45)mL,hospital stay(5.5±1.5)days,urethral catheterization time(12.6±1.8)days,and no patient received blood transfusion during operation.The urinary continence rates at the time of catheter removal,and at 1,3,and 6 months after surgery were 39.1%,65.2%,73.9%,and 91.3%,respectively.Two patients had positive margins,both of which were at the neurovascular bundle.No patient developed surgery-related complications,urinary obstruction or fistula after surgery.Conclusion Vesicourethral anastomosis with MULP and BNP in LRP for HRPC can effectively improve patients'early urinary continence rate and postoperative quality of life without increasing the oncological risk.
2.Comparison of three kinds of mesenchymal stem cells differentiating into nerve cells under co-culture induction
Lili XU ; Hongyuan WANG ; Xueda LI ; Bing LIU ; Fangfang ZHENG ; Nailong YANG
Chinese Journal of Tissue Engineering Research 2017;21(17):2714-2721
BACKGROUND: Scholars have been trying to create a microenvironment similar to the human body, which can induce the directional differentiation of mesenchymal stem cells from human bone marrow, placenta and umbilical cord blood. OBJECTIVE: To compare the neuronal differentiation of human bone marrow mesenchymal stem cells, human placental mesenchymal stem cells and human umbilical cord blood mesenchymal stem cells induced by co-culture with nerve cells. METHODS: Human bone marrow mesenchymal stem cells, human placental mesenchymal stem cells and human umbilical cord blood mesenchymal stem cells cultured in vitro were co-cultured with nerve cells using the Transwell system. The morphological changes of three kinds of cells in the co-culture system were detected. After co-culture for4-5 days, immunofluorescence staining was used to measure the expression of neuron-specific enolase in cells. Mesenchymal stem cells only cultured in low glucose DMEM medium were used as controls. RESULTS AND CONCLUSION: These three kinds of mesenchymal stem cells were extended, and interconnected processes were detective. The positive expression of neuron-specific enolase was highest in the human umbilical cord blood mesenchymal stem cells followed by human placental mesenchymal stem cells and human bone marrow mesenchymal stem cells in order. In the control group, none of the three kinds of mesenchymal stem cells have neuronal morphology, and the expression of neuron specific enolase was negative for the immunofluorescence staining. To conclude, microenvironment provided by nerve cells can induce these three kinds of mesenchymal stem cells todifferentiate into neurons.
3.One case of nonketotic hyperglycemia hemichorea
Lili XI ; Liyan SHEN ; Qing WANG ; Ping WANG ; Liping LIU ; Nailong YANG
Chinese Journal of Endocrine Surgery 2016;10(6):525-526
4.Uric acid effect on Wnt signaling pathways during osteogenic differentiation of human bone marrow mesenchymal stem cells
Xiaoli WANG ; Lili XU ; Nailong YANG
Chinese Journal of Tissue Engineering Research 2015;(28):4472-4477
BACKGROUND:Uric acid, as an endogenous antioxidant, exhibits anti-oxidative and anti-DNA damage effects, promotes osteogenic differentiation, and therefore has been paid more attentions.
OBJECTIVE:To investigate the effects of different concentrations of uric acid on the expression of genes related to Wnt/β-catenin signaling pathways during osteogenic differentiation of human bone marrow mesenchymal stem cel s.
METHODS:Healthy adult human bone marrow mesenchymal stem cel s were in vitro isolated by adherent culture of whole bone marrow. Passage 3 bone marrow mesenchymal stem cel s were induced to differentiate into osteoblasts using different concentrations of uric acid (0, 140, 280, 560μmol/L). Alkaline phosphatase activity, cel proliferation capacity, and Wnt-3αandβ-catenin mRNA expression in the Wnt signaling pathways were detected.
RESULTS AND CONCLUSION:Alkaline phosphatase staining was positive. After treatment with uric acid, alkaline phosphatase activity and cel proliferation capacity were increased, the expression of Wnt signaling pathway-related genes Wnt-3a andβ-catenin was up-regulated in a dose-dependent manner. There were significant differences in the abovementioned indices between experimental groups and between each experimental group and control group (P<0.05). These findings suggest that uric acid up-regulates Wnt-3a/β-catenin signaling pathway and thereby promotes the osteogenic differentiation and proliferation of human bone marrow mesenchymal stem cel s in a dose-dependent manner.
5.Osteogenic induction of human bone marrow mesenchymal stem cells cultured in complex medium
Lili XU ; Xiaojuan SUN ; Xiuxian HAO ; Tingting XIE ; Nailong YANG
Chinese Journal of Tissue Engineering Research 2015;(10):1501-1505
BACKGROUND:Studies have shown that the number of osteoblasts is often decreased after osteoporosis, and osteoblast replacement therapy becomes a new target for the treatment of osteoporosis. OBJECTIVE:To observe the osteogenic differentiation of human bone marrow mesenchymal stem cels cultured in dexamethasone, vitamin C and beta-glycerophosphate. METHODS:Mesenchymal stem cels were isolated and purified from adult bone marrow using human lymphocyte separation medium. The expression of cel surface markers was detected by flow cytometry. Cel ultrastructure was observed by transmission electron microscope. Then, the bone marrow mesenchymal stem cels were cultured in osteogenic induction medium containing dexamethasone, vitamin C andβ-glycerophosphate, and RT-PCR was used to detect the bone morphogenetic protein-2 mRNA expression after osteogenic induction. RESULTS AND CONCLUSION:A large number of adherent cels were visible as fibrous growth at 2 weeks after culture and strongly expressed CD44, CD29, but did not express CD34, CD45. These cels could be induced to differentiate into osteoblasts, and express bone morphogenetic protein-2 mRNA. Alizarin red staining and alkaline phosphatase staining were positive for the cels. These findings suggest that human bone marrow mesenchymal stem cels cultured in dexamethasone, vitamin C and beta-glycerophosphate can differentiate into osteoblasts, and has a potential for the treatment of osteoporosis.
6.Effect of different concentrations of uric acid on the neural differentiation of human bone marrow mesenchymal stem cells
Qingqing SONG ; Lingfan YU ; Lili XU ; Nailong YANG
Chinese Journal of Tissue Engineering Research 2014;(6):847-852
BACKGROUND:Uric acid as an endogenous antioxidant has garnered increasing attentions because of its anti-oxidation, anti-DNA damage and neuroprotective effects.
OBJECTIVE:To observe the effect of uric acid at different concentrations on the neural differentiation of bone marrow mesenchymal stem cells.
METHODS:Bone marrow mesenchymal stem cells were isolated, purified and cultured in vitro. The morphology change was observed. The third passages of bone marrow mesenchymal stem cells were induced to differentiate to neuron-like cells by induced liquid containing four different concentrations of uric acid (0 mmol/L as control group, 0.2 mmol/L, 0.4 mmol/L, 0.8 mmol/L) for 24 hours. Then, after second intervention for 1 hour, cells were detected by Nissl staining and specific markers were detected by immunohistochemistry method.
RESULTS AND CONCLUSION:After induction, the cellbody shrank, forming processes and connections. Nissl body was found in the cytoplasm. The positive rates of neuron-specific enolase were significantly higher in uric acid groups of different concentrations compared to the control group (P<0.05);moreover, the positive rates of neuron-specific enolase were increased as the increase in concentrations of uric acid (P<0.05). The positive rates of Nestin were decreased in uric acid groups of different concentrations compared to the control group (P<0.05). After 4 hours of induction, cells fel off significantly. In a certain period of time, uric acid can promote differentiation of bone marrow mesenchymal stem cells into neuron-like cells in a certain concentration-dependent manner in vitro.
7.The investigation and evaluation of bone mineral density in type 2 diabetic women
Nailong YANG ; Jun WANG ; Ning QU
Chinese Journal of Diabetes 2008;16(1):26-28
Objective To observe the bone mineral density(BMD)in type 2 diabetic women and discuss the methods for evaluation of BMD.Methods BMD was measured by dual-energy X-ray absorptiometry(DEXA)at skeletal regions of supine and lateral spine,left femur neck and left hip in 484 type 2 diabetic women and 868 healthy women.The relation of BMD of healthy women in skeletal regions with age changes was found to fit a cubic regression model.Results The peak BMD of lateral spine and hip appeared at 25-34 and 35-44 years old respectively.BMD in diabetic women was not different from age-matched healthy women.The BMD values of the different age stages in diabetic women showed no difference(P>0.05)as compared with healthy women,and decreased with the age increase.The numbers of damaged bone sites were increased with the age increase.The BMD difference values between peak value and values of different age stages showed no difference between diabetic and healthy women groups(P>0.05).Conclusion The changes of BMD in type 2 diabetic women show no difference from age-matched healthy women.And in both diabetic and healthy women,BMD decreases with age increasing.
8.Sources, induced differentiation and molecular markers of pancreatic stem cells
Chinese Journal of Tissue Engineering Research 2007;0(07):-
OBJECTIVE: Pancreatic stem cell transplantation is a new approach for diabetes treatment. It may overcome the shortage of human donor islets and avoid the long-term applying of immunity depressors. In this article, we review the recent progress of the sources, induced differentiation and the molecular markers of pancreatic stem cells. DATA SOURCES: A computer-based online search of PUBMED was undertaken to identify relative articles published in English from June 1997 to June 2006 with the keywords of "pancreatic, stem cell, differentiate, marker". Meanwhile, we searched Wanfang database for the related Chinese articles published between June 1997 and June 2006 with the same key words in Chinese. STUDY SELECTION: The articles were firstly selected and the quotations cited by every article were looked over. Inclusive criteria: The articles focusing on the source, induced differentiation and molecular markers of pancreatic stem cells. Exclusive criteria: Repeated experiments or Meta analysis articles. DATA EXTRACTION: According to the inclusive criteria, 31 articles were finally selected among the 82 ones collected from databases.Fifty-one old data or duplicated researches were excluded. Among the 31 selected articles, 6 presented the main conceptions, 15 were related to the sources of pancreatic stem cell and its induced differentiation, 9 were focused on the surface molecular markers and 2 discussed the major obstacles at present. DATA SYNTHESIS: Stem cells are pluripotential cells with the ability to self-renew; they are classified into two major classes: embryonic stem cell and adult stem cell. Pancreatic stem cells, belonging to the adult stem cells, could differentiate into tube cells, insulin-secreting cells, exocrine cells and other specific pancreatic tissue cells, which possess the fundamental properties of long-term proliferation and self-renewal. The studies on the sources, induced differentiation and molecular markers of pancreatic stem cells are extremely useful for the treatment of diabetes. Recent studies have shown that besides gene engineering, insulin-secreting cells are mainly generated from the differentiation of embryonic stem cells and adult stem cells such as pancreatic duct epithelial cells, nestin-expressing pancreatic stem/precursor cells, bone marrow mesenchymal stem cells and haematopoietic stem cells. The investigation on the molecular markers of pancreatic stem cell surface is essential to the basic and clinical research. PDX-1, nestin, CK-19, CK-20, Ngn3 and PGP9.5 are the major markers used in the identification, isolation and purification of pancreatic stem cells. CONCLUSION: The investigations on islet cells and pancreatic stem cells transplantations are promoting. Significant advancements in search for the sources and identification of pancreatic stem cells have been achieved. However, many obstacles should be overcome before any clinical breakthroughs can be expected. With the significant improvement in stem cell research and techniques, it is possible to obtain enough pancreatic stem cells from in vitro culture for transplantation.
9.Comparison between two isolation methods of human mesenchymal stem cells in vitro
Chinese Journal of Tissue Engineering Research 2007;0(03):-
AIM:There is no standard isolation method of mesenchymal stem cells.Percoll density gradient centrifugation is treated as the traditional method among numerous methods but it is so complicated.This study compares whole bone marrow culture to traditional Percoll density gradient centrifugation in order to establish a better method in vitro.METHODS:Experiments were conducted at Central Laboratory of Affiliated Hospital of Qingdao University Medical College from September 2006 to June 2007.Bone marrow was supported by patients who had autologous stem cell transplantation by endocrinology department for diabetes treatment.The experiments had known by the patients and authorized by the hospital ethic committee.The two different methods,whole bone marrow culture and Percoll density gradient centrifugation,were compared in the number of cell clone,cell morphology,cell superficial mark,the differentiation into fat cell.RESULTS:① The number of clone cell by whole bone marrow culture was more than by Percoll density gradient centrifugation(P
10.Differentiation of adult bone marrow mesenchymal stem cells into islet-like cells in vitro
Chinese Journal of Tissue Engineering Research 2007;0(08):-
BACKGROUND: In recent years, there are some effects on using pancreatic islet cell or pancreas transplantation in the treatment of diabetes. But there are still two major problems: lack of donor and immune rejection. Mesenchymal stem cells can be induced to islet-like cells in vitro. OBJECTIVE: To study the progress in the differentiation of insulin secretion cells induced by mesenchymal stem cells. RETRIEVAL STRATEGY: A computer-based online search of Pubmed was undertaken for the related English articles dated from August 1997 to August 2007 with the of "mesenchymal stem cells,insulin secreting cell". At the same time, we searched China Journal Full-Text Database for the related Chinese articles published between August 1997 and August 2007 with the same key words. Total retrieved 108 literatures which are firstly selected into the inclusive criteria: ①The articles focusing on the mesenchymal stem cells induced differentiation into insulin secreting cells. ②choose articles at the same field or published in authoritative magazine published in the recent time. Exclusive criteria: ①Repeated experiments or ②Meta analysis articles. LITERATURE EVALUATION: All the literatures are about the differentiation of insulin secretion cells induced by bone marrow mesenchymal stem cells. Of the 30 selected literatures, 2 are reviews and the others are clinical or basic experiment study. DATA SYNTHESIS: ①Because the pancreas is similar to the nervous system in the mechanism of development and control, now people think that nerve growth factor may be the key signal to the pancreas development. Mesenchymal stem cells have an ability of multi-differentiation; it can be induced into nestin positive cells by using epidermal growth factor and basic fibroblast growth factor. Mesenchymal stem cells will be further induced to pancreatic cells if we add conditioned medium as the appropriate micro-environment for islet cells. ②Now identification and functional study of insulin-like cells are for the following areas: observe whether there is islet-like cells group gathered by cell morphology; test insulin, glucagons and somatostatin expression after induction; test insulin and glucagons gene expression using RT-PCR method after induction; to study the function of induced pancreatic cells by using glucose-stimulated insulin release test. CONCLUSION: As the research about mesenchymal stem cells can be induced islet-like cell in vitro becoming ever-changing technology, it becomes a new program of cell therapy to diabetes in face of all problems. According to all the present experimental results, the insulin secretion of induced pancreatic islet cells is much smaller than the normal amount of insulin secretion. Therefore, how to induce efficiently with human physiology of islet cell in vitro and then transplant safely into the human body will bring a new leap of diabetes treatment.

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