1.In vitro chondrocyte differentiation of canine bone marrow stromal stem cells
Qiyou WANG ; Yichun XU ; Daozhang CAI
Chinese Journal of Tissue Engineering Research 2009;13(46):9037-9040
BACKGROUND:Bone marrow stromal stem cells(BMSCs)do not allow for single differentiation of chondrocytes due to their multi-directional differentiation,bone morphogenetic protein secreted from osteoblasts affect the non-differentiated precursor cells and promote their osteoblast differentiations,while those differentiated cells are bound to form tissues.OBJECTIVE:To in vitro induce canine BMSCs differentiate into chondrocytes,and to investigate the method and conditions of chondrocyte differentiation in vitro.DESIGN,TIME AND SETTING:Single sample observation was performed in the Laboratory of Tissue Engineering,Sun Yat-sen University between March 2005 and January 2006.MATERIALS:One male dog,aged 4 months,was involved to harvest BMSCs from the rib.METHODS:Rib BMSCs extracted from bone marrow of 2.0-3.0 mL were cultured in vitro. When cells reached a confluence at 8-11 days,trypsinization was conducted and then halted with L-DMEM synthesis culture solution containing 10% fetal bovine serum. Cellular suspension was collected and centrifuged,cells were rssuspended and incubated at a ratio of 1:3. The third generation of cells were cultured and amplified,10 μg/L basic fibroblast growth factor 2 mL was added to replenish culture medium twice,then 1 mg/L transforming growth factor β1 of 2 mL was applied to induce BMSCs differentiation into chondrocytes.MAIN OUTCOME MEASURES:Toluidine blue and alcian blue stains were applied to determine cartilage matdx secretion,immunohistochemistTy was used for the detection of cartilage specific Ⅱ collagen expression.RESULTS:After BMSCs were primarily cultured and subcultured in vitro,they were shown to grow well at the fourth generation,those induced by basic fibroblast growth factor and transforming growth factor β1 were positive for toluidine blue and aician blue staining;immunohistochemistry showed a positive outcome for type Ⅱ collagen,indicating the induced BMSCs exhibited chondrocyte's characteristics.CONCLUSION:Utilizing basic fibroblast growth factor and transforming growth factor β1,the induced canine BMSCs could differentiate into chondrocytes,which is considered as an ideal seed cells for cartilage tissue engineering.
2.A study on the natural history of hematuria associated with benign prostatic hyperplasia and the effect of finasteride
Yuanli WANG ; Qiyou CAO ; Huai YANG
Chinese Journal of Urology 2001;0(10):-
Objective To study the effect of finasteride on hematuria associated with benign prostate hyperplasia. Methods We evaluated 61 patients with intermittent hematuria who were randomized to a finasteride treated group or control group.Routine urinoscopy was carried out once a week in all the 61 patients. Results In the untreated control group hematuria recurred in 20/31 within a year but in only 7/28 in the finasteride group,which was a statistically significant difference (P
3.Adrenal incidentalomas:clinical analysis of 52 cases
Yuanli WANG ; Weilie HU ; Qiyou CAO ; Wei WANG ; Xiaoming ZHANG
Chinese Journal of Urology 2001;0(07):-
Objective To improve the diagnosis and differential diagnosis of adrenal incidentalomas for the optimal management of adrenal masses. Methods The clinical data of 52 cases (35 men and 17 women;mean age,42 years;age range,23-65 years) of adrenal incidentalomas was reviewed.Among them, 33 cases had tumors ≥ 3.0 cm in diameter.None of 52 cases had clinical symptoms and signs of hypertension and endocrine dysfunction.All of the tumors were incidentally found by ultrasonography or CT on regular physical examination.A series of blood chemical examinations (including electrolytes,adrenal cortical and medullary metabolic biochemistry),abdominal ultrasound and CT were performed in all 52 cases;and MRI,PET and 131 I-MIBG imaging were performed in 6,2 and 1 cases,respectively.Surgery and postoperative pathological study were performed in all. Results Postoperative pathology showed adrenocortical adenoma in 27 cases,adrenal metastatic carcinoma in 7,adrenal pheochromocytoma in 5,adrenal cyst in 4,primary aldosteronoma in 3,adrenal neurinoma in 2,adrenocortical carcinoma in 1,retroperitoneal neuroblastoma in 1,retroperitoneal primary lymphoma in 1 and adrenal ganglionic fibroma in 1.Diagnoses based on the results of imaging examinations and laboratory tests were correct in 14 cases (27%).Of the 10 patients with malignancies,5 with primary diseases died 6-10 months after chemotherapy and 5 were lost to follow-up.The remaining 42 patients were followed up for 6 months to 9 years with no evidence of recurrence. Conclusions Multidisciplinary approaches are required to identify and differentiate adrenal incidentalomas, and early surgical treatment is suggested.
4.Biocompatibility of poly-3-hydroxybutyrate-co-3-hydroxyvalerate and canine bone marrow stem cells in vitro
Qiyou WANG ; Yichun XU ; Limin RONG ; Daozhang CAI
Chinese Journal of Tissue Engineering Research 2009;13(16):3045-3048
BACKGROUND: Poly-3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV) is a novel scaffold made by solvent casting/particulate leaching procedure, composed of polyhydroxybutyrate and polyhyclroxyvalerate at certain ratio, which has good biocompatibility as well as high intensity and modulus. It has three-dimensional porous net structure and good biodegradation. OBJECTIVE: To evaluate the biocompatibility between copolymers of PHBV and canine bone marrow mesenchymal stem cells(BMSCs).DESIGN, TIME AND SETTING: In vitro comparative observation. The study was performed at the Laboratory of Histology and Embryology, Sun Yat-sen University between June 2003 and March 2004.MATERIALS: PHBV scaffold, film porosity > 85% and 100-350 μ m aperture size.METHODS: Canine BMSCs were isolated and cultured. The 3-4 passage cells were seeded onto the PHBV films and three-dimensional foam scaffold. Cells cultured alone served as control.MAIN OUTCOME MEASURES: The seeded cells were observed under inverted microscope; at 1, 2, 3 weeks after seeding, the BMSCs were treated with 4% paraformaldehyde and stained with hematoxylin-eosin (HE); The protein content in seeded cells was determined by bicinchoninic acid assay (BCA), and the content of DNA was quantified using Hoechst33258 assay at 5, 10, 14 days after culture.RESULTS: Inverted microscopic observation showed that the PHBV fibers were fairly thick with weak lucency, and the fibers were hardly detectable under contrast phase microscope. Majority of cells attached onto the PHBV films 2 hours after seeding, and extended well in a spindle shape at 3 days. One week after culture, 2 PHBV were fixed, and BMSCs proliferation was observed after HE staining. At two weeks, cells continued to proliferate and densely covered the PHBV film. The cells grew in the three-dimensional pores, connected at 1 week, extended at 3 weeks, secreting a large amount of material around cells. Cell proliferation did not change much at 3 weeks compared with 2 weeks, and there was no significant difference in DNA and protein contents between control and PHBV groups (P > 0.05).CONCLUSION: As a kind of tissue-engineered scaffold material for BMSCs, PHBV displays good biocompatibility.
5.Clinical characteristic and perioperative management of colonic obstruction caused by colonic carcinoma in elderly
Qiyou SHEN ; Bin WANG ; Chunxin ZHANG ; Qinzhi LIU
Chinese Journal of Postgraduates of Medicine 2012;35(2):17-19
ObjectiveTo discuss the clinical characteristic and perioperative management of colonic obstruction caused by colonic carcinoma in elderly.MethodsThe clinical characteristic and perioperative management of 30 patients with colonic obstruction caused by colonic carcinoma from January 2000 to November 2010 were collected and analyzed retrospectively.Among 30 cases,11 cases were accepted emergency operation,19 cases were accepted limited operation.Resection and anastomosis at stage Ⅰ was in 26 cases (86.7%),shortcut surgery and simple fistulation was in 4 cases ( 13.5% ).ResultsThe length of stay was 22-56 days.Postoperative complication including wound infection in 5 cases,wound liquescence in 3 cases,wound dehisce in 2 cases,anastomotic fistula in 2 cases,pulmonary infection in 3 cases.Two cases died during perioperative period.Conclusions Besides diagnosis as early as possible,appropriate perioperative management should be taken to the elderly patients,including reasonable operative time and surgical approaches and these are the keys to improve clinical curative effect.
6.Clinical study of laparoscopic cholecystectomy combined with endoscopic retrograde cholangiopancreatography/endoscopic Oddi sphincterotomy on the treatment of cholecystolithiasis complicated by choledocholithiasis
Qiyou SHEN ; Chenggang JI ; Bin WANG ; Chunxin ZHANG
Chinese Journal of Postgraduates of Medicine 2011;34(14):24-26
Objective To discuss the clinical value of laparoscopic cholecystectomy (LC) combined with endoscopic retrograde cholangiopancreatography (ERCP)/endoscopic Oddi sphincterotomy (EST) on treating choledocholithiasis with cholecystolithiasis. Method The clinical data of 48 patients with choledocholithiasis complicated by cholecystolithiasis treated with LC combined with ERCP/EST from January 2005 to August 2010 was collected and analyzed retrospectively. Results Forty-six patients achieyed success by ERCP,and 45 patients finished LC,all patients underwent LC combined with ERCP/EST were recovered more rapidly,with shorter hospital stays. No severe complications or residual stone and refluent cholangitis in the follow-up of 3-12 months of 46 patients. Conclusions Combining the advantages of LC with ERCP/EST treating patients with choledocholithiasis complicated by cholecystolithiasis,according to the theoretics of minimally invasive surgery, with less invasive and the advantages of shorter hospital stays and rapid recovery. It is the comparatively ideal choice for the treatment of choledocholithiasis complicated by cholecystolithiasis at present.
7.Clinical use of fast-track surgery in colorectal surgery
Qinzhi LIU ; Qiyou SHEN ; Chunxin ZHANG ; Bin WANG
Chinese Journal of Postgraduates of Medicine 2011;34(3):17-19
Objective To investigate the feasibility of fast-track surgery in colorectal surgery.Methods Eighty consecutive patients with colorectal cancer admitted for colorectal surgery were divided into two groups by random digits table with 40 cases each. Group A was treated with the new concept of fasttrack surgery and group B was treated with the traditional methods of operation. The time of postoperative bowel venting and defecation,hospital stay time, the rate of complication, the rate of readmission and the total cost during hospitalization were compared. Results The time of postoperative bowel venting and defecation,hospital stay time were shorter in group A [(2.7 ± 0.9), (2.9 ± 0.1 ), (5.8 ± 1.0) d,respectively]than those in group B [( 3.9 ± 0.5 ), (4.2 ± 0.3 ), ( 8.3 ± 1.2) d, respectively] and the rate of complication and the total cost during hospitalization in group A [7.5%(3/40), ( 1.83 ± 0.22) ten thousand yuan] were lower than those in group B[27.5%(11/40), (2.35 ± 0.36) ten thousand yuan](P< 0.05). Conclusion The new concept of fast-track surgery can accelerate recovery after colorectal resection,reduce the rate of overall complications and total cost during hospitalization.
8.Establishment of finite element model of lumbar motion segment implanted with artificial lumbar intervertebral disc and its stress analysis
Yichun XU ; Shangli LIU ; Meichao ZHANG ; Daozhang CAI ; Qiyou WANG
Chinese Journal of Tissue Engineering Research 2005;9(26):240-242
BACKGROUND: At present, there are very big differences in structure,material character and biological property between artificial intervertebral disc (AID) and normal physiological intervertebral disc.OBJECTIVE: Three-dimensional finite element method was used to observe and analysis the stress conduction of artificial lumbar intervertebral disc in lumbar motion segment.DESIGN: Single sample observation was designed.SETTING: Department of Orthopaedics, Third Affiliated Hospital, Sun Yat-sen University; Department of Orthopaedics, Second Affiliated Hospital, Sun Yat-sen University; Laboratory of Mechanics, Southern Medical UniversityPARTICIPANTS: It was to employ a vertebral sample without any spinal disorder of a healthy male died due to accidence and a finite element model of AID implantation in vertebral motion segment established with SB Charite Ⅲ AID.METHODS: According to industrial design chart of AID, finite element software MSC.MARK was utilized to establish three-dimensional model of artificial lumbar intervertebral disc. The corpus sample of motion segment of healthy lumbar vertebrae was collected and scanned with spiral CT machine and imaging documents were input in computer to preserve.Geometric model of L4-5 segment was established in three-dimensional coordinate system in ASC.MARK software. The intervertebral disc in L4-5 motion segment model was replaced by AID. It was to ensure the fixation of lower terminal lamina of L5 in the model. 4 Nm moment of force was exerted in anterior flexion, posterior extension, lateral bending and torsion on the sample successively. Finally, force of internodes representing AID was calculated and stress distribution was recorded.MAIN OUTCOME MEASURES: To observe stress distribution of anterior flexion, posterior extension, compression, lateral bending and rotation of AID.RESULTS: Finite element model of artificial lumbar intervertebral disc implanted lumbar motion segment that is in conformity with clinical practice was established. Stress distribution of AID was characterized as:er lamina was the maximum and that in the lower inclined part of slide of slide core and cover lamina was two or three times as same as that of sion, the stress in the center of slide core and cover lamina was the maximum.CONCLUSION: The finite element model of artificial lumbar intervertebral disc implanted lumbar motion segment established is in conformity with the structural character of practical artificial intervertebral disc in morphology, size and motion property, based on which, it is feasible to carry on the experiment on stress distribution of artificial intervertebral disc.
9.Comparison of surgery and conservative treatment for senile dens fractures of Anderson-D Alonzo type Ⅱ or Ⅲ: a Meta-analysis
Huajun LING ; Lei FAN ; Jianwen DONG ; Weiwen LIN ; Qiyou WANG
Chinese Journal of Orthopaedic Trauma 2017;19(3):225-233
Objective To compare the operative and conservative treatments for senile dens fractures of Anderson-D Alonzo type Ⅱ or Ⅲ using Meta-analysis.Methods A literature search was conducted in PubMed,Embase,Web of Science,Cochrane Library,Wanfang Data and CNKI for studies on senile dens fractures of Anderson-D Alonzo type Ⅱ or Ⅲ from the earliest records through June 2016.The relative studies identified were screened by 2 independent authors.The quality of these articles was evaluated using modified Newcastle-Ottawa scale,and the meta-analysis was conducted using Review Manager 5.3.Results A total of 22 articles were brought into this Meta-analysis.The union rate was significantly higher in the operative group than in the conservative group[OR =0.30,95% CI(0.20,0.44),P < 0.001];the mortality in the operative group was significantly lower than in the conservative group[OR =0.61,95% CI (0.39,0.96),P=0.03];the complication rate was similar in both groups[OR=1.09,95% CI(0.76,1.57),P < 0.46].The heterogeneity of all the 3 indexes was low.In the subgroup analysis,the union rate was significantly higher in posterior operations than in anterior operations or conservative treatments (P < 0.05).Conclusion For the elderly patients with dens fracture of type Ⅱ or Ⅲ who can tolerate surgery,operative treatment may be more suitable because it can lead to much better prognosis.
10.In vitro cartilage tissue engineering with bone marrow stromal cells and photografting modified copolymers of 3-hydroxybutyrate and 3-hydroxyvalerate
Qichun ZHAO ; Daozhang CAI ; Qiyou WANG ; Bin LIU ; Zhusheng CHEN
Chinese Journal of Microsurgery 2008;31(4):-
Objective To explore the feasibility of building tissue engineered cartilage by bone marrow stromal cells and pbotografting modified copolymers of 3-hydroxybutymte and 3-hydroxyvalerate.Methods Sheep BMSCs were seeded in three-dimensional photografting modified PHBV scaffoids.Twenty-four hours later.composites were cultured with ehondrogenically inductive medium(DMEM)containing TGF-B(10 ng/m1),IGF-1(150 ng/m1)and 20% fetal bovine serum.Three weeks later,the constructs were evaluated by scanning electron microscopy(SEM)and light microscopy with alcian blue,safrine 0 and type Ⅱ collage immunohistochemical staining.GAG contents of constructs were determined by DMB(1,9-dimethylmethylene blue)binding assay at weekly intervals up to 3 weeks.The composites were implanted subcutaneously in sheep abedoml and were evaluated macroscopically and bistologically at 4 weeks postoperatively.Results SEM photograph showed.after one week culture,cell morphology changed from fibroblast-like elongated spindle to the flat rounded like chondrocytes,and the extra cellular matrix also increased obviousl~.Furthmore,with the culture time extension,this change were more evident.HE staining showed that cells filled all the inter-connected pores in the constructs.And more cells were observed in the outer layer of the constructs.ECM(extraeellular matrix)Was strongly positive by Aleian blue,Safrine O staining and type Ⅱ collage immunohistechemical staining.DMB binding assay revealed that the induced BMSCs GAG secretion(1306.7±192.3)wag significantly higher than BMSCs(205.0±26.2)(P<0.001),but it was significantly lower than passage 2 ehondrocytes(1969.2±235.3)(P<0.001).Saltine O and type Ⅱ collage immunohistochemical staining were positive in constructs implanted subcutaneously.Conclusion Tissue engineered cartilage could be obtained using BMSCs and photografting modified PHBV,but there are still gaps physiologically between the constructs and the nature cartilage.