0.05), but were highly significant at doses of 100 ?mol/L or above (P

BACKGROUND:Rapid development of spermatogonial stem cells is the new hope for assisting reproductive technologies,and the stability of the number and structure of the chromosome cultured in vitro is one of the important factors in usage.OBJECTIVE: To study the influential factors on G-band exhibition in mouse spermatogonial stem cells, so as to offer a technology to identify karyotype of stem cells in culture.DESIGN: Observational experiment.SETTING: Luzhou Medical College.MATERIALS: The experiment was conducted in the Laboratory of Medical Molecular Biology, Luzhou Medical College from March 2004 to April 2005. 10 days old male and female Kunming mice were provided by Department of Animal of Luzhou Medical College (number of license No. 17 of experimental animal quality administration in Sichuan province).There were low-sugar DMEM medium, 10 mg/L Mitomycin-C, IMDM medium, 1 ×10-5 mol/L colchicines (PBS allocation),7.5 mmol/L KCL, fixative (mixture glacial acetic acid with methanol in 1:3), Giemsa staining solution and 0.25% zymine.METHODS: Bone marrow was aspirated from the thigh bone of mouse for feeder layer cells preparation. Cells from the male mice testis of 7-8 days after birth were prepared and made into cell suspension. After adjusting the cell density to 3×105 L-1, they were inoculated into the feeder layer of bone marrow stromal cells. Cells were cultured at 37 ℃ in CO2 incubator containing CO2 of 0.05 volume fraction and 70% humidity. The proliferation groups of stem cells cultured 15-20 days were selected, stirred and spread, and then treated with colchicine for 4-6 hours. Cell suspension was collected,and then stained after hypotonic treatment. The cells whose chromosomes were dispersed well and in metaphase were selected, and number of chromosomes was counted, and then the morphology of chromosomes was observed.MAIN OUTCOME MEASURES: Culture of bone marrow stromal cells and spermatogonial stem cells and coloration and count of chromosomes.RESULTS: The karyotype of spermatogonial stem cells was the same as the body cells of normal mouse, which showed granule or rod-shape. Karyotype was 20 pairs, 40 bars. Three kinds of chromosome morphology could be observed under oil immersion lens. The first type was condensed, which could be counted in total, but the band could not be seen.The second type was chromatosome that spread completely in the center of equatorial plate and were in metaphase. In this phase, total numbers and band were seen clearly. Last type-chromosomes had already folded and moved towards two poles and concentrated gradually, the total number of chromosomes could be counted, but bending could not be seen clearly.CONCLUSION:Many factors can affect the karyotype of spermatogonial stem cells, including the phase of cell division,effect of hypotonic solution, diffusion of the cell when dropping slides, concentration of trypsin and digestion time, etc.

Objective To summarize the experience in endovascular repair of De Bakey type Ⅲ aortic dissection in recent years and summarize the prevention and management of the related perioperative complications.Methods From January 2009 to January 2011,49 cases of endovascular repair for De Bakey type Ⅲ aortic dissection were performed under general anesthesia in our department.There were 45 male and 4 female.The follow-up was performed in the outpatient department or by telephone.Results There was no inhospital death and no paraplegia events.Severe complication included:coma,2 cases ( 4.1％ ) ; endoleak,2 cases (4.1％ ) ; upper limb ischemia,2 cases (4.1％ ).Recurrent proximal aortic dissection,1 case.Fever was occurred in most of those cases.Conclusion Endovascular repair of aortic dissection improves the outcome of aortic dissection patients.But more attention should be pay to prevent the severe complications,It will help to improve the prognosis and life quality by reducing the risk of retrograde dissection,acute brain ischemia and endoleak.

Objective:To explore the personnel arrangement in the outpatient pharmacy by calculating working hour to provide ref-erence for the rational staffing in hospital. Methods:The daily work content and working hour of 18 pharmacists in the outpatient phar-macy of a large general hospital from January to March in 2013 were following-up observed and recorded using the working hour meas-urement. The data were input the EXcellsoftware to establish the database, and the workload in various positions was collected and sorted. The obtained relative parameters were used to calculate the needed worker number on the basis of manpower planning model. Results:The research confirmed the mean operation time for 9 work programs in the outpatient pharmacy, and the time for drug dispen-sing and distributing was detailed. The needed number of pharmacists was 13. 29 according to the calculation, plus the officer-in-charge and sanitation workers, the total number was 15. 29(approx. 16). Conclusion:The working hour measurement can scientifically de-termine the time for each job, and the workload should be used as the foundation for configuring personnel qualification and the number in outpatient pharmacy.

Objective To investigate and evaluate the curative effect of radioimmunological targeting drug on nude mice bearing breast cancer. Methods The anti-CEA monoclonal antibody C50 was combined with ~ 131 I to produce radioimmunological targeting drug. 16 nude mice inoculated subcutaneously with breast cancer cell MCF-7 with tumor diameter about 0.5 cm were randomly into 4 groups(n=4): group Ⅰ, injected in part with ~ 131 I-C50 18.5 MBq; group Ⅱ, injected in part with ~ 131 I-C50 3.7 MBq; group Ⅲ, injected in part with ~ 131 I-mIgG 18.5 MBq; group Ⅳ, injected in part with C50 0.75 ?g. The size of tumor volume and inhibitory rate (IR) after treatment for six weeks were calculated and compared with the control group. Results The tumor volume and curves for tumor growth and tumor weight had significant differences between group Ⅰ and the group Ⅲ as well as group Ⅱ (P0.05). Conclusion Anti-CEA monoclonal antibody C50 labeled with radionuclide ~ 131 I could inhibit the growth of the tumor when given locally. ~ 131 I-C50 has a potential value of clinical application

Objective To explore the related risk factors of renal function after partial nephrectomy,and provide the reference for clin-ic. Methods The data of 31 patients with small renal cancer in our hospital from January 2010 to June 2014 were analyzed. Before and after the treatment,the relationship of renal glomerular filtration rate ( GFR) with the change and clinical pathological factors were analyzed,the single factor analysis used t test,multiple factors analysis adopted multi-factor unconditioned logistic regression analysis. Results Single fac-tor analysis results showed that renal function after partial nephrectomy was closely related to the age,diabetes mellitus,blocking time,the size of the tumor,renal volume reduction ratio,the difference was statistically significant (P<0. 05). Many factors of logistic regression showed that it was closely related to the age,blocking time,renal volume reduction ratio(P<0. 05). Conclusion The age,blocking time,renal vol-ume reduction ratio are risk factors for renal function after partial nephrectomy.

Objective To explore the clinical effect of the treatment for fresh achilles tendon rupture by modified double Kessler suture with S type incision .Methods Totally 21 cases of fresh achilles tendon rupture were admitted into our hospital from January 2012 to Decem-ber 2015, and they were treated by S type incision modified double Kessler suture within 7 days (averagely 4.1 days).Observed the postop-erative complications and evaluated the efficacy with Arner-Lindholm.Results All the 21 patients were followed for 6 to 24 months ( aver-agely 12 months).There was no incision infection ,necrosis, re-fracture of achilles tendon and other complications .There were 16 cases of excellent and 5 cases of good , with the excellent and good rate of 100% through the Arner-Lindholm evaluation criteria .Conclusion Modified double Kessler suture with S incision for the treatment of fresh achilles tendon rupture is conducive to early functional exercise ,and it can obtain satisfactory clinical efficacy .

Objective To comfirm the anti-fibrotic effect of spironolactone and investigate the influence of spironolactone on concentration of angiotensinⅡ(AngⅡ) in rat hepatic fibrosis. Methods Ninety male Sprague-Dawly rats were randomly divided into three groups: control group (8 rats, subcutaneous injection of pure peanut oil, normal diet), model group (42 rats, subcutaneous injection of 40% CCl 4, 0.3 mL/100 g, twice a week, high cholesterol and low protein diet, 5% alcohol as the drink), and spironolactone group (40 rats, same dosage of CCl 4, high cholesterol and low protein diet, gastropufusion of spironolactone at the dose of 100 mg?kg -1?d -1). The animals were sacrificed at 2, 4, 6 and 8 weeks of administration. AngⅡ was assayed by radioimmunoassay. The liver tissues were stained, coded and examined. Results ① The general conditions of the rats of spironolactone group were better than those of the model group. The degree of fibrosis and the area of collagen in the liver of spironolactone group were less than those of the model group after the 4th week (P

Objective To summarize our experience in endovascular repair of 83 cases with type B aortic dissection.Methods A retrospective analysis was performed in 83 cases of type B aortic dissections who were treated in our hospital.Results The surgical procedure was successful for all the patients.Two patients died peri-operatively.One case encountered a coma,but no post-operative paraplegia occurred.Conclusion Endovascular repair for type B aortic dissection is a micro-invasive,safe and effective technique.Long-term follow-up is required to give a comprehensive evaluation.

Objective To determine whether store-operated Ca2+ entry (SOCE) is involved in chronic hypoxia-induced alteration of intracellular Ca2 + concentration ([Ca2+] i) and proliferation in pulmonary arterial smooth muscle cells (PASMC).Methods Rat PASMCs were cultured and treated in normoxia (21％O2) or hypoxia (4％ O2) condition.The proliferation of PASMC was detected by cell counting kit-8 (CCK-8) assay.[Ca2 +] i,SOCE and the effects of store-operated Ca2 + channel (SOCC) inhibitors,SKF96365 and NiCl2,on SOCE in hypoxic PASMCs were tested by InCyte [Ca2 +] i measurement system.Results Hypoxia for 24-60 h augmented PASMC proliferation (1.12 ± 0.09 vs 0.71 ± 0.05,P ＜ 0.05) and [Ca2 +] i [(214.8 ± 20.4) nmol/L vs (115.2 ± 13.2) nmol/L,P ＜ 0.05] in a time-dependent manner with the maximum effect at 60 h.Perfusion of Ca2+-free Krebs solution containing nifedipine (5 μ mol/L),cyclopiazonic acid (CPA,10 μmol/L) in PASMCs caused a small transient increase of [Ca2+]i with peak [Ca2+]i (113.3 ± 49.3) nmol/L.Chronic hypoxia (4％ O2,60 h) enhanced [Ca2+]i level with peak value of (193.2 ± 22.7) nmol/L (P ＜ 0.05) in PASMC.After restoration of extracellular Ca2+,CPA caused marked increase of [Ca2+]i with peak value of (328.0 ± 56.7) nmol/L.Chronic hypoxia strengthened CPA-induced increase of [Ca2 +] i with peak value of (526.0 ± 33.7) nmol/L (P ＜ 0.05) in PASMCs.Either SKF96365 50 μmol/L or NiCl2 500 μmol/L distinctly attenuated CPA-induced enhancement of [Ca2 +] i,the peak value of which dropped from (526.0 ± 33.7) nmol/L to (170.4 ± 26.4) nmol/L (P＜0.05) or (177.4±45.9) nmol/L (P＜0.05) respectively.Conclusion Chronic hypoxia boosts the release of Ca2+ from sarcoplasmic reticulum and promotes the activity of SOCC and SOCE,leading to [Ca2 +] i elevation and proliferation of rat PASMCs.