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.

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.

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.

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.

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.

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.

BACKGROUND: There are no studies on differentiation of bone marrow mesenchymal stem cells (BMSCs) into neurons directly induced by neural cells. OBJECTIVE: To establish the co-culture system between BMSCs and neural cells in vitro, and to study the influence of neural cells on the differentiation of BMSCs into neuron in the co-culture system. DESIGN, TIME AND SETTING: The in vitro cytology control experiment was performed at the Central Laboratory of Hospital Affiliated to Medical College of Qingdao University from December 2006 to December 2007. MATERIALS: BMSCS were harvested from bone marrow of patients with diabetes meliuts, who underwent autologous stem cell transplantation at the Hospital Affiliated to Medical College of Qingdao University. Neural cells were collected from brain tissues of infants die of asphyxia during delivery. The third passage of neural cells was used in this study. Transwell double-deck culture dish was purchased from Corning Costar, with a pore diameter of less than 3.0 ?m. Cells could not traverse, but the medium could traverse. METHODS: Neural cells were incubated in Transwell double-deck culture dish at a density of 1?106 in the co-culture group. BMSCs were incubated in the upper layer in LG-DMEM medium for 4-5 days. BMSCs were incubated in both layers in the control group. MAIN OUTCOME MEASURES: The morphological changes of BMSCs were observed and the special markers of neurons cells in BMSCs were examined by immunofluorescence. RESULTS: BMSCs in the co-culture group grew slowly, showing radial processes and connected each other. Positive rate of neuron specific enolase was (32.7?11.5)%. BMSCs in the control group were flat and wide, and negative for neuron specific enolase. CONCLUSION: Microenvironment provided by neural cells promotes the differentiation of BMSCs into neurons.

BACKGROUND: Cells grew slowly in a low number after conventional induction. Growth factor can accelerate induction speed, but growth factor is expensive. OBJECTIVE: To verify the efficiency of differentiation of human bone marrow mesenchymal stem cells (BMSCs) into osteoblasts in vitro in a inductor without growth factor. DESIGN, TIME AND SETTING: The control cytology observation experiment was conducted from July 2007 to February 2008 at the Center Laboratory of Hospital Affiliated to Medical College of Qingdao University. MATERIALS: Bone marrow was collected from diabetic patients undergoing autologous stem cell transplantation. Informed consent was obtained from patients. METHODS: Using density gradient method, BMSCs were separate and primary cultured. At the third passage, BMSCs were incubated in 0.1 ?mol/L dexamethasone, 10 mmol/L ?-glycerine sodium, and 50 ?mol/L vitamin C. BMSCs in the control group were left intact. MAIN OUTCOME MEASURES: Morphology of BMSCs was observed under a phase contrast microscope. Morphological changes were measured by alkaline phosphatase staining and calcified nodules staining after induction. RESULTS: Under the phase contrast microscope, the third passage of BMSCs was uniform, spindle. At 7 days, cells were confluence. Some cells were overlapped over time. Mesenchymal tissues gradually piled up, with mineralization, multiple nodes. At 14 days, brown mineralized nodes were found under an inverted microscope. At 21 days, mineralization in fragmentis were formed. Von Kossa method demonstrated black calcified nodules and positive alkaline phosphatase. CONCLUSION: BMSCs can be induced into osteoblasts easily, with high and rapid proliferation.

BACKGROUND: There is no ideal method about adult bone marrow mesenchymal stem cells (BMSCs) differentiate into islet-secreting cells and appreciation in vitro at present. Transgene requests strict conditions and complex program. The induction using chemicals is a present-used method. OBJECTIVE: To investigate the culture conditions for inducing adult BMSCs into islet-like cells in vitro. DESIGN, TIME AND SETTING: The cytology in vitro experiment was performed at the Central Laboratory of Affiliated Hospital of Medical College of Qingdao University from January to October 2007. MATERIALS: Bone marrow was collected from diabetic patients undergoing autologous stem cell transplantation at Department of Endocrinology, Affiliated Hospital of Medical College of Qingdao University after signing the informed consent. Epidermal growth factor and basic fibroblast growth factor were obtained from Peprotech Asia. B27 adjunct was purchased from Gibco. Nicotinamide, 2-mercaptoethanol, glutamine and dithizone were bought from Sigma, USA. METHODS: The adult BMSCs were isolated by Percoll from adult bone marrow aspirates and cultured in LG-DMEM, were suspended by Trypsin and passaged for subsequent passages. At the third to fifth passages, BMSCs were incubated at a density of 1?108 L-1 and were induced differentiation into islet-like cells through three developmental stages. In the first stage, BMSCs were incubated in HG-DMEM supplemented with 2-mercaptoethanol, glutamine for 2 days. In the second stage, BMSCs were incubated in HG-DMEM supplemented with epidermal growth factor, basic fibroblast growth factor, B27 and glutamine for 6 days. In the third stage, BMSCs were incubated in HG-DMEM supplemented with nicotinamide and 2-mercaptoethanol for 6 days. BMSCS in the control group were only incubated in the HG-DMEM. MAIN OUTCOME MEASURES: Morphological changes in BMSCs were analyzed under a phase contrast microscopy. Duodenal Homeobox 1 (PDX-1) expression was detected during the second and differentiation stage by immunofluorescence assay. Islet ?-like cell clusters from the 3rd stage were identified by positive dithizone staining. The insulin secretions under the stimulation of high or low glucose were detected by electrochemiluminescence immunoassay. RESULTS: The undifferentiated BMSCs exhibited adherent long spindle-shaped cells. After induction, cells gradually became round and formed clusters. Cells expressed the PDX-1 gene at 8 days and formed islet-like cell clusters that exhibited positive dithizone staining at 14 days. After high and low glucose treatment, no insulin was detected in the control group; insulin content significantly increased at 14 days (t=3.638-9.387, P