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Background:Serum pepsinogens (PGs),as a serologic marker for gastric mucosal lesions,can reflect the functional status of gastric mucosa. Helicobacter pylori (Hp)infection can cause pathological changes in gastric mucosa,and has been reported to influence the serum level of PGs. Aims:To explore the influence of Hp infection on diagnostic performance of serum PGs for gastric mucosal lesions. Methods:The endoscopic findings,biopsy pathology (including Giemsa staining) and serum PGs test in 1216 cases of patients from July 2014 to June 2015 at the First Affiliated Hospital of Jiaxing University were collected. Patients were categorized according to the pathological diagnosis and Hp status,and the results of serum PGs test were analyzed between different groups. Results:When patients were classified by gastric mucosal lesion,no significant differences were found in serum levels of PGⅠ,PGⅡ,ratio for PGⅠ/ Ⅱ (PGR)and proportion of PG-positive (PGⅠ≤70 μg/ L and PGR < 3. 0)patients of different mucosal lesion groups with Hp-positive status (P >0. 05),whereas significant differences were observed in serum levels of PGⅠ,PGⅡ,PGR and proportions of patients with PGⅠ≤70 μg/ L or PGR < 3. 0 of different mucosal lesion groups with Hp-negative status (P < 0. 05). When patients were classified by Hp status,PGⅠ level and PGR were lower and PGⅡ level and proportion of PG-positive patients were higher in Hp-positive patients than in Hp-negative patients in any of the gastric mucosal lesion groups (P < 0. 05 in part of the comparisons). Conclusions:Hp infection is strongly associated with the alterations in serum PGs test,which narrows the differences in PGs between groups with different gastric mucosal lesions and expands that within the same mucosal lesion, subsequently decreasing PGR and increasing the proportion of PG-positive patients. Patients negative for Hp infection may need new cut-off value of serum PGs test to improve the sensitivity for diagnosis of gastric mucosal lesion.
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ObjectiveTo investigate the effects of using spiral pedunculated bladder muscle flap ureteroplasty in the treatment of long ureteral segment defects ( ＞ 20 cm).MethodsA retrospective analysis was conducted on the clinical effects of five patients who encountered long ureteral segment defects caused during ureteroscopic lithotripsy.The five patients included three males and two females with an age range from 37 to 59 yrs ( average age 48 ).Four of the cases had defects on the left and one case on the right.Two cases had whole ureteral mucosal avulsion and three cases had whole ureteral ruptur from the pelvis to the bladder junction.Defect lengths measured from 21 to 25 cm( mean length 22.5 cm).All five patients underwent emergency surgery using spiral pedunculated bladder muscle flap ureteroplasty and 7 F double J stent placement in the repaired ureters which was fixed on psoas muscles.The average length of the new ureters using spiral pedunculated bladder muscle flap was 22.5 cm.ResultsAll the operations were successful and the operation time was 1 -2 hrs (average 1.5 hrs).Drainage tubes for four patients were removed three days after operation.IN the remaining case the drainage tube was removed 10 days after surgery due to urine leakage.All wounds healed uneventfully.Serum creatinine and blood urea nitrogen were normal two weeks after surgery.Double-J tubes were removed safely under cystoscope eight weeks after surgery.In following-up,one case was found to have mild hydronephrosis and ipsilateral ureter slight expansion six months after surgery,but renal function was normal.There was no abnomality found in the remaining four patients after 2 -4 years of follow-up.The IVU showed normal morphology and good developments in the ipsilateral ureter.ConclusionsSpiral pedunculated bladder muscle flap ureteroplasty is an ideal treatment method in repairing long ureteral segment defects.
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Objective To explore the underlying clinical factors and precautionary measures of fluid extravasation in patients with calyceal calculi treated by ureteroscopic holmium laser lithotripsy.Methods A retrospective review was made on clinical records of 138 patients with calyceal calculi receiving retrograde ureteroscopic holmium laser lithotripsy from May 2005 to March 2009. The relevance was studied between the occurance of fluid extravasation complications and various clinical factors using x2 test and binary Logistic regression. The clinical factors included patients' sexes, age groups (＜30 years, 30-50 years, ＞50 years), history of treatment (ESWL or open surgery) for upper urinary tract calculi, preoperative upper urinary tract infection, intraoperative placement of ureteral catheter and the length of procedure duration (＜ 50 min, 50-80 min, ＞ 80 min). Results Fluid extravasation complications occurred in 24 patients. The sexes and age groups were irrelevant to the occurance of fluid extravasation complications; while history of ESWL or open surgery and preoperative infection in upper urinary tract, without intraoperative ureteral catheter placement and long duration of procedure were responsible for the higher rates of the fluid extravasation complications.Conclusion Reasonable selection of patients and timing of operation, regular intraoperative ureteral catheter placement and control the length of procedure duration help to reduce fluid extravasation during retrograde ureteroscopic lithotripsy.
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Objective To study the effect and histocompatibility of urethral extracellular matrix in repair of traumatic urethra defects in rabbits. Methods Model of traumatic urethral defects was made by resecting 1.0-1.5 cm segment of the urethra in 20 rabbits. Then, the defects were repaired by a tube of extracellular matrix of the same length. The dynamic changes of CD4+, CD8+ T cell and CD4+/CD8+ in peripheral blood were detected by flow cytometry at 1,2, 3 and 4 weeks after operation. In the meantime, the immunity response of rabbits was evaluated by lymphocyte transformation test. The repaired segments stained with hematoxylin-eosin (HE) and Van Giesen were studied by histologic and pathologic observations at 10 days, 3, 6 and 24 weeks postoperatively. The urodynamics, urethroscopy and urethrography were performed at 24 weeks postoperatively. Results There was no significant difference in aspects of stimulative index of lymphocyte transformation, T lymphocyte subsets CD4+, CD8+ T cell and CD4+/CD8+ between experimental group and negative control group. Urothelium covered the whole surface of the matrix tube three weeks after operation. The smooth muscle cells increased nearly to normal urethral wall at 24 weeks. Urethrosoopy and urethrography showed glossy matrix tube. There was no statistical difference upon urodynamies between experimental group and control group. Conclusion The urethral extracellular matrix has good histocompatibility and may be a safe and effective material for repairing urethra defects.
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Objective To detect the mouse testicular gene expression pattern differences be-tween spermatogonial stem cell (SSC) proliferative and differential stages and study the molecular reg-ulation mechanism in SSC proliferation and differentiation. Methods With the interval of 24 days, male Kunming mice were injected intraperitoneally with two doses of busulfan (10 mg/kg) to establish spermatogenesis regeneration models. 36 k Mouse Genome Array was used to detect the differential gene expression profiles between the stages of SSC proliferation and differentiation. Bioinforrnsties analysis was conducted in GO (gene ontology) and KEGG (Kyoto Encyclopedia of Genes and Ge-nomes) pathway to describe the potential roles that may play in spermatogonial stern cells behavior regulation. Results Nine hundred and eleven differential expression genes were identified by gene arrays in mice testes, consisting of 608 up-regulated and 303 down-regulated in SSC proliferation stage and SSC differentiation stage. The differential expression genes were classified by their biological process, molecular function and cellular component, respectively. Alterations with statistical signifi-cance (P<0.05)appeared in 84 KEG(;signal pathways, including Notch and Wnt signaling pathways which had been proved to be important for stem cell maintenance. Fifty-six differential expression genes were selected as genes related to stem cells, among which 40 genes were up-regulated, including some stem cell biomarkers(such as Cd9, StraS, hgbl-, Oct4 and Thyl)and some growth factors(such as Fgf2, Pdgfa and Csfl). Conclustion The regulation of SSC proliferation and differentiation involves inmany differentially expressed genes in various signal pathways. This study provides a molecular basis for the elucidation of the molecular mechanism behind self-renewal and differentiation of spermatogonial stem cells.
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Objective To establish a long-term culture system for mouse spermatogonial stem cells(SSCs). Methods Testis cells from 4-6 days postpartum male transgenic BALB/C mice were collected by a modified two-step enzymatic digestion method.After three differential adherence selections,the enriched germ cells were finally suspended in StemPro-34 SFM medium supplemented with other nutrients factors and plated on mouse embryonic fibroblast(MEF)feeder layer.20 ng/ml Glial cell line-derived neurotrophic factor,10 ng/ml basic fibroblast growth factor and 200 ng/ml GDNF-family receptor al were added to the serum-free medium to promote SSCs proliferation.Aduh male BALB/C mice,4-5 weeks old,underwent intraperitoneal injection of 40 mg/kg busulfan as recipient mice.Cultured SSCs were also injected into the seminiferous tubules of the left recipient testis through micromanipulator and right testis as self-control.Testes of recipient mice were observed by a fluorescence stereomicroscope and HE stains at 2 months after transplantation. Results By improved digestion method,the vitality of isolated testis cells was more than 98%and the stem cells was enriched about 18.5 fold. 1-2 days after transferred to MEF feeder, the round germ cells started to proliferate and had the shape of paired or aligned undifferentiated spermatogonia connected by cytoplasmic bridges. After 3-4 days, SSCs proliferated continuously and formed typical colonies. SSCs from BALB/c mice could be cultured and passaged in a steady state for 3 months. Cryostat section through the transplanted testis showed that most of seminiferous tubules were filled with germ cells expressing EGFP.HE staining further showed clearly that seminiferous tubules contained complete spermatogenesis.Conclusions SSCs from BALB/c mice could be cultured in an improved culture system for 3 months.The culture system could facilitate understanding the regulatory mechanism that governs SSCs and might provide an opportunity for the cure of infertility.
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BACKGROUND: Vessel extracellular matrix (VECM) is a natural scaffold material obtained from vascular tissues, which can stimulate angiogenesis and accelerate vascularization of tissue-engineered graft, however, the mechanism is poorly understood.OBJECTIVE: To explore the vascularization effects of release of vascular endothelial growth factor (VEGF) from VECM in ureteral reconstitution.DESIGN, TIME AND SETTING: An in vitro cytology observation. The experiment was performed at the Biomedical Engineering Laboratory of First Clinical Medical Science College, Wuhan University, between April and August in 2009.MATERIALS: Abdominal aorta was obtained from 5 rabbits to prepare VECM.METHODS: The VEGF released from VECM in vitro was evaluated by ELISA, the effects of cell proliferation by the released VEGF was detected by MTT colorimetric assay. The defected ureters of rabbits were repaired by homologous VECM in vivo.Then the recovery of the defected ureters and the situation of vasculogenesis were detected at different time point.MAIN OUTCOME MEASURES: The detection of VEGF contents in VECM; and the effects of VECM on vascular endothelial cell proliferation and ureteral reconstitution.RESULTS: In vitro experiment presented that the peak amplitude concentration of VEGF released from VECM in PBS solution was (124.10±1.42) ng/L, which showed proliferative effect on vascular endothelial cells. In vivo, there were some blood vessels on the VECM at 2 weeks after implantation. Epithelial coverage was evident in the lumen of the marginal part of the VECM grafts and the smooth muscle extended from the transition zone. After 8 weeks, the quantity of the blood vessel was increased and the caliber of the blood vessels became wide. There was thickness epithelial lamina in the graft, and the muscle fibers had an organized spatial alignment, forming variably sized bundles. After 16 weeks, there were no significant differences between the regenerative tissue and the normal tissue in morphology.CONCLUSION: The homologous VECM can release VEGF when implanted as tissue engineer biomaterial and might be an ideal replacement biomaterial for ureteral reconstitution.
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BACKGROUND:It is accepted that the best method for spermatogonial stem cells separation is using artificial cryptorchism model combined with surface makers.OBJECTIVE:To explore the feasibility of separation spermatogonial stem cells with ?6-integrin and c-kit as specific surface markers.DESIGN,TIME AND SETTING:The randomized control experiment was performed at the Renmin Hospital of Wuhan University from May to December 2006.MATERIALS:Forty adult,white Kunming mice with 6 weeks old were randomly divided into cryptorchidism and control groups,with 20 animals in each group.METHODS:Artificial cryptorchidism model was prepared by made an incision at the median of abdomen,and testis was pulled into abdominal cavity,which was fixed at the each side of lateral abdominal wall.There was no treatment in the control group.The single cell suspension of seminiferous epithelium was obtained by traditional two step enzyme digestion at 2-3 months after operation.FITC-conjugated anti-?6-intergrin antibody and PE-conjugated anti-c-kit antibodies were added.Then the cells with low side scatter light-scattering properties were sorted and positively stained for ?6-intergrin and negative c-kit expression.Meanwhile,the viability of the isolated cells was assessed by trypan blue staining.MAIN OUTCOME MEASURES:The morphological changes of cryptorchidism,and the sorting results of spermatogonial stem cells.RESULTS:Cell distribution in seminiferous tubule was disorder with reduced numbers.The layer and lumens were disappeared,and cell division phase could be seen in the center of tubules.Compared to the control group,the testicular cells in the cryptorchidism group were increased in the side scatterlow,Forward scatterhi areas,with figure left-upward displacement.The distribution of ?6-integrin+ and c-kit cells were deviated each other,it named that most ?6-integrin+ cell were not spermatogonial stem cells,so do the c-kit-cells.Only 2.8% of testicular cells exhibited side scatterlow,?6-integrin+,and c-kit-,which were spermatogonial stem cells in the cryptorchidism group.And trypan blue staining showed that over 95% of them were viable.CONCLUSION:Using the two surface markers to sort spermatogonial stem cells can advance the purity of the spermatogonial stem cells in cell suspension,but the specificity is insufficient.
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BACKGROUND: Skeletal muscle-derived stem cells (MDSCs), another adult pluripotent stem cells, have become a hot topic in the field of gene therapy and cell-based tissue engineering. It has wide prospect in treating stress incontinence by periurethral injection of patients' MDSCs.OBJECTIVE: To investigate the method of culturing MDSCs in vitro so as to provide experimental basis for treating stress incotinence by injecting MDSCs.DESIGN: Repeated observation.SETTING: Department of Reproductive Medical Center and Department of Urology, Renmin Hospital of Wuhan University.MATERIALS: This experiment was conducted at the Laboratory of Department of Urology , Renmin Hospital , wuhan University from December 2003 to May 2004. Totally 20 female SD rats , aged 4 to 6 weeks , were involved . Type- Ⅺ pancreatin, collgenase , pancreatin ,polylysine (Sigma), dispase enzyme (Gibco) , chick embryo extract (self made).METHODS: ① Rats were anesthetized intraperitoneally with 10 g/Lpentobarbital sodium (30 g/kg). Under aspetic condition, gastroeneminus were isolated and immediately put into pre-cooled DMEM media (Gibco)containing antibiotics and then removed into the hood. After washed with D-hank's solution three times, muscle biopsies were removed of fascia,tendon, nerve and blood vessels and minced into small pieces about 1-3 mm3,and then transferred into a centrifugation tube. 0.2% collgenase-type Ⅺ(Sigma) and 0.1% pancreatin were added to the left tissue . Skeletal muscle cells of the rats were isolated with collagenase. ② Differential attachment was used to purify the skeletal muscle cells of the rats. After screened with cell screen cloth , cell suspension was transferred into a polylysine-coated flask and cultured at 37 ℃ in a humid atmosphere with 0.05 CO2 in air for 1 hour. All the cells that did not adhere to the flask were then transferred to another flask with new culture medium and cultured for approximately 1 hour at 37 ℃ .Again,the non-adhering cells were transferred to another flask and were incubated at 37 ℃ overnight.The technique was carried out for an additional 4-5 days. This operation was repeated every 24 hours until the fifth day. Those cells attached on days 5-6 were MDSCs.③ After they grew with 70% confluence , the cells were digested with trypsin and passaged at the ratio of 1:2. After digestion, the generative cells were inoculated to the culture plate with 6-well cover glass. Growing culture medium was added and cell slide was prepaared 24 hours later. The expression of specific protein antigen and stem cell antigen-1 of the cultured cells were identified with immunohistochemical staining.MAIN OUTCOME MEASURES: Morphology and identification of MDSCs.RESULTS: ① Morphology of MDSCs: Cells isolated from skeletal muscle tissue took the form of spheres and had high refraction. When subcultured,they began to adhere at the 12th hour, at that time they were still round,and complete their attachment at the 48th hour. Then they began to expand and ultimately became fusiform-shaped or spindle-shaped, having two poles and small size. As time going, they fused with each other to form mature poly-nuclear myotubes. ② Identification results of muscle-derived stem cells: MDSCs were desmin and stem cell antigen-1 (Sca-1) positive specific for some stem cells. Red fluorescence effluenced from cytoplasm was found under the fluorescence microscope and the positive rate reached 90%.CONCLUSION: MDSC belongs to adult stem cells and has the advantages of rich source, low immunogenicity and long survival after transplantation. High-purity muscle-derived stem cells can be obtained through primary culture , and immunohistochemical technique can identify muscle-derived and specific characteristics of stem cells, so it has a broad application perspective in tissue engineering and gene therapy.
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This is an experiment on rabbits to evaluate the possibility of ureteral replacement by extracellular matrix. We adopted a biochemical method for preparing a new tissue engineering material named Extracellular Matrix (ECM), and the ECMs were used as homologous grafts to replace the defect in the ureters. Light microscopy, scanning electron microscopy, immunohistochemical technique and intravenous urography were used. The routine blood and biochemical laboratory tests were made before and after operation, and the measured values of pressure in the ureter of experiment and control groups were compared. The ureteral ECM was found in the experiment to promote the regeneration of all ureteral wall components. There were no significant differentces between the regenerative tissue and the normal tissue in morphology and function 16 weeks after replacement. The homologous ECM might be an ideal replacement material for ureteral defect.
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Animals , Female , Male , Rabbits , Biomedical Engineering , Methods , Bioprosthesis , Epithelium , Physiology , Extracellular Matrix , Physiology , Transplantation , Random Allocation , Transplantation, Autologous , Ureter , Wounds and Injuries , Pathology , General SurgeryABSTRACT
Objective To investigate the protective mechanism of cyclosporine A to renal autografts in situ.Methods Thirty rabbits were divided into three groups at random (n=10 in each group), subjected to left renal in situ perfusion with normal saline (control group), HC-A solution (HC-A group) and HC-A solution containing CsA (30 ml/L, experimental group) respectively. By ~using self-made in situ cryopreservation device, the left kidney was cryopreserved in situ at 0-4 ℃ for 2 h. Then the right kidney was dissected and the left renal blood flow was opened. At 6th h and 24th h after reperfusion, blood BUN and Cr levels were determined. At 24th h, the left kidneys were ~dissected for the detection of the expression of HSP70 and NF-?B in left renal tissues. The apoptotic rate in left renal tissues was assayed by in situ end-labeling method. Results In the experimental group, the expression level of HSP70 was significantly higher, while the expression of NF-?B and ~apoptotic rate significantly lower than in the other two groups (all P
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Objective To obtain a simple method to isolate and purify spermatogonia stem cells. Methods The testis of newborn mouse were digested by modified two step digest method, and the type A spermatogonia cells were separated by different adhesive time with Sertoli cells. Results High concentration (95%) purified type A spermatogonia cells were acquired, and the spermatogonia transplanting experiment demonstrated the efficiency of the method.Conclusions This method is easy and efficient to isolation and purification of spermatogonial stem cells.
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Objective To investigate the feasibility of an ileal reservoir with self controllable urination. Methods A segment of ileum was under taken,the middle portion of which was made into a N shaped reservoir and the proximal portion used as the afferent limb.The distal ileum was pulled out and the two skin flaps were taken and encircled the efferent ileum to constitute the urinary outflow.A urinary controller similar to penile clamp was applied to control the urination with a balloon catheter. Results Ten dogs and five patients underwent this procedure and the patients were followed up for 3 to 14 months.Urodynamic examination showed that on the third month,the reservoir in animals reached a maximum capacity of (150?40)ml and a full filling pressure of (24.4?5.3)cmH 2O.The reservoir in the patients reached a maximum capacity of (290?80)ml,a full filling pressure of (36.3?8.2)cmH 2O,a maximum urinary flow rate of (20.3?4.7)ml/s with no residual urine.X ray examination showed good excretory function of the kidney without hydronephrosis.All patients can control urination themselves through the use of the urinary controller. Conclusions The new procedure has the advantages of self control lable of urination,simple surgical performance and few complications.
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Objective To improve the surgical technique for the removal of huge staghorn stones. Methods 202 cases treated with an incision of the intrarenal sinus supplemented by a postrenal lower pole segmental incision for the removal of huge kidney stones were studied. Results The biggest stone was 9.8 cm?5.0 cm?3.2 cm and in one case the stones amouned to 74. The average postoperative hospitalization days was 12.8. Postoperative examination with KUB+IVU or B ultrasound revealed residual stone in 9 cases, the diameters being less than 0.8 cm. Conclusions The procedure needs no interruption of renal pedical and with the advantages of simple,less bleeding,complete removal of calculus,and preservation of renal function.
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0.05). Compared with preoperation, serum LH levels were significantly higher after operation in both groups(P0.05). DSA and color Doppler ultrasound examination showed that autografts had good blood supply. Conclusion The modified testicular autotransplantation operation could improve the successful rate of transplantation, and had not obvious influence on testicular function. It can be further applied in homologous testicular transplantation experimental and clinical research.
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Objective To evaluate the effect of incision of the intrarenal sinus plus postrenal low pole segmental incision for removal of calculus on the renal hemodynamics. Methods The hemodynamic changes of intrarenal and incisional sites in 98 cases with complex calculi were consecutively monitored by color Doppler ultrasonograph before operation and also 1 month,3 months,6 months after operation.The ratio(S/D) of the peak flow-rate in systole(S),the flow-rate at the end of diastole(D),and the resistant index(RI) of the intrarenal chief renal arteries and intersegmental arteries and interlobar arteries were analyzed.The signals of blood stream in segmental incision were divided into four classes(Ⅰ to Ⅳ) and were observed. Results The changes of the intrarenal hemodynamics were related to the degrees of hydronephrosis(P0.05),but it caused the blood stream to be obstructed in the area of incision where the signals of blood stream showed classes Ⅲ to Ⅳ.The patients generally recovered after 3 to 6 months,and the time of recovery was associated with the degrees of hydronephrosis Conclusions It is concluded that the incision of the intrarenal sinus plus postrenal low pole segmental incision for removal of calculus is an effective and safe method for treat-ment of complex renal calculus.
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Objective To investigate the value of the clinical application of fetal testis for the treatment of male hypogonadism.Method We have performed the testicular transplantation to cure 6 patients suffered from hypogonadism with the fetal testis as donor.Results All patients had a significantly increased level of serum testosterone and the male secondary sexual characteristics and the sexual desire were improved.The size of testis was larger than before operation.Conclusion Testis transplantation with fetal testis as donor is an effective method to cure male hypogonadism and has great value of clinical application for its weak immunogenicity.
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Objective To study a new procedure of surgical repairment of rectourethral fistula.Method By the perineum approach one stage urethroplasty using island skin tubes from perineum and scrotum has been adopted after the resection of fistular urethra and repairment of the defect of the anterior wall of rectum to substitute the defect of urethra in 9 cases during 1992-1998.Result The 9 cases were all cured and have been followed-up for 2 6 years without recurrence of rectourethral fistula.Conclusion This procedure was regarded as an ideal technique for the closure of rectourethral fistula especially for those complicated with urethra stricture.
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Objective To investigate the value of the clinical application of fetal testis transplantation with main vessel segment. Methods The testicular transplantation were performed to cure 9 patients suffered from male hyponadism with the fetal testis with main vessel segment as donor tissue. Results Except one, 8 patients had a significantly increased level of serum testosterone, the male secondary sexual characteristics and the sexual desire were improved. The size of testis was larger than before operation. Conclusion Fetal testis transplantation with main vessel segment is a effective method to cure male hyponadism and has great value of clinical application.
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Objective To evaluate the sickle renal parenchyma incision for the removal of complex staghorn renal calculi. Methods Sickle parenchyma incision was used to remove stones in 37 patients with complex staghorn renal calculi.The procedure was as follows:the kidney was disected free and the pelvis within sinus renalis was isolated.Two rows of botton style sutures were made on the renal parenchyma with 2-0 plain catgut along mid-lower 1/3 of the dorsal surface of kidney free of vessels (Brodie's line) from the renal posterior lib to the plane of lower major calyx.The renal parenchyma was opened.Then,the incision was developed from the plane of lower major calyx through the middle major calyx to the plane of upper major calyx.The shape of this incision appeared like a sickle.The renal parenchyma and each calyx along this incision were opened and so was all the stones could then be easily removed.The calyces could well be observed. Results The calculi were completely removed in all the 37 cases.21 needed intraoperative blood transfusion and the mean amount of blood was 120 ml.KUB+IVU were normal 4 weeks postoperatively with improved hydronephrosis,no intrarenal stricture and void of residual calculi. Conclusions This procedure has the advantages of little intraoperative bleeding,slight impairment of renal function,high clearence rate and is indicated for the removal of any intrarenal pelvis complex staghorn calculi.