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Objective To test and analyze the genetic background of highly immunodeficient mice from different sources.Methods Four highly immunodeficient mouse strains from different sources of NOD background were collected. 30 microsatellite DNA sites were detected, and the genotype can be displayed by gel electrophoresis and STR scanning. Results 17 microsatellite sites exhibit polymorphism in 20 mice of the four groups.There were 30 homozygous loci in the mice of groups A and B, and heterozygous in the other two groups.The genetic distance is minimum between groups A and B, showing a higher genetic similarity.Conclusions The genetic backgrounds are different in highly immunodeficient mice from different sources.
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Objective To improve the gene targeting efficiency with C57BL/6 embryonic stem ( ES) cells.Meth-ods Three different genetically modified C57BL/6 ES cell lines, named TLX3, Ai3K and SL, were microinjected into ICR, B6( Cg)-Tyrc-2J and BALB/c mouse blastocysts, respectively.The efficiency was statistically evaluated according to three aspects:blastocyst collection, chimera production and germline transmission.Results None of the three ES cell lines was germline transmitted with B6(Cg)-Tyrc-2J mice as blastocyst donors, while it was achieved with both BALB/c and ICR mouse blastocysts.Compared in the aspect of blastocysts collection, ICR mouse was much better than BALB/c mouse (P<0.05), and the chimera production efficiency of ICR mouse was comparable to that of BALB/c mouse (P =0.115). As to the germline transmission efficiency, that of BALB/c mice is significantly higher than that of the ICR mice ( P<0.01).Conclusions The germline transmission efficiency of BALB/c mouse is highest among these three mouse strains. However, it has the disadvantages of blastocyst collection, developmental delay and zona pellucida fragility, compared with ICR mouse.Therefore, ICR mouse is also a good candidate as blastocyst donor for embryonic stem cell microinjection.
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Objective To obtain the basic growth parameters of a self-established F1 hybrid CB6F1 C57-ras transgenic mouse model, and to provide basic information for commercialization of this mouse model. Methods F1 hybrid mice (CB6F1) were produced by crossing C57-ras heterozygous transgenic (c-Ha-ras+/-) male mice and wild-type BALB/cJ female mice.The average litter size, weaning rate, sex ratio, growth performance and C57-ras transgenic positive rate were recorded and analyzed.Results The average litter size was eight, weaning rate was 90%, and sex ratio was approximately in accordance with prediction.The average body weight of newborn mice was 1.73 ±0.05 g.The average body weight of 10-week old c-Ha-ras transgenic female and male mice in CB6F1 background was 24.38 ±1.74 g and 29.42 ±1.72g, respectively, which had a significant difference (P<0.01).The c-Ha-ras transgenic positive rate was 46.9%. which was in accordance with genetic rules.Conclusions The F1 hybrid mice (CB6F1) produced by crossing C57-ras heterozygous transgenic ( c-Ha-ras +/-) male mice and wild-type BALB/cJ female mice show normal growth performance and development characteristics, and it can be used for large-scale commercial supply.
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Objective To discuss the effect of in vitro fertilization ( IVF) and mouse sperm cryopreservation , to establish a simple and economic frozen system for the genetically engineering mice preservation .Methods Sperm from genetically engineering mice were cryopreserved , IVF was performed using post-thawed sperm, then embryo transfer, to compare the effects of cryopreservation medium、age of male mice and sperm preincubation medium .Results Using CPA as sperm cryopreservation medium , when PM was used thawed-sperm preincubation in IVF , the fertility rates were from 82.49%to 91.43%, when HTF was used thawed-sperm preincubation in IVF , the fertility rates were from14.46%to 27.38%, there was a signification difference between PM and HTF sperm preincubation medium;10 to 35 weeks male genetically engineering mice sperm were succeed cryopreservation , and positive mice were procreated after 2-cell embryos were transferred;R18S3、CPM and CPA was used to freeze sperm , the fertility rates were 75.85%、88.89%to 94.27%, positive mice were procreated after 2-cell embryos were transferred;2-cell embryos after IVF were freezed , then thawed and positive mice were procreated after 2-cell embryos were transferred .Conclusion Using CPA as sperm cryopreservation medium , when PM was used thawed-sperm preincubation in IVF , genetically engineering mice sperm were succeed cryopreservation .
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Vitrification provides a rapid cooling to induce glass-like solidification inside cells and protect cell membrane and cytoskeleton system free of injury by ice crystals.The main factors that can influence the effect of vitrification are the size of ovarian tissue, the kind of cryoprotectants, the method of permeation, carrier system and so on. Ovarian tissue structure is very complex, and there is no uniform standardized protocol of vitrification yet.This paper presents also the current problems of ovarian tissue vitrification.
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Objective To establish a simple , fast and economic total DNA extraction method to serve the rapid identification of model mouse genotype in large number of mice .Methods Three methods, i.e.phenol extraction, isopropyl alcohol precipitation and mouse ear boiling methods were used to extract the total DNA from ten C 57-rasmodel mice.The purity and yield of DNA obtained by the three methods were compared , and polymerase chain reaction ( PCR) assay was used to compare the efficacy of the three extraction methods .Results Among the three methods , phenol extraction was the best and isopropyl alcohol precipitation was the poorest in DNA yield .In terms of DNA purity , the phenol extraction was the best and the mouse ear boiling method was the poorest .All the three methods could be used to extract the total DNA from mice serving as template of PCR reaction for the mouse genotype identification .The time consumption of three methods are 12.5 hr ,13 hr and 0.18 hr.Mouse ear boiling method was significantly lower than that of the other two methods ( P <0.01 ) ,.The obtained total DNA can be stored at conventional -20℃for 7 days and 30 days later still can be used as a template for PCR reaction .Conclusions Among the three methods studied , the mouse ear boiling method is simple and with the lowest cost , so it is feasible for total DNA extraction in scaled genotyping experiments .
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@#Objective To explore the diagnosis and treatment of supinator syndrome.Methods 11 patients with supinator syndrome definitely diagnosed were treated with conservation treatment, 4 ineffective cases with conservation treatment were treated with operation. The following-up was performed for 6～12 months to evaluate the therapeutic effect.Results Among the total 7 patients received conservation treatment, 5 cases recovered completely, the other 2 cases recovered partially; among the total 4 patients who received operations, 3 cases recovered completely, and the other one recovered partially.Conclusion At present, supinator syndrome can be diagnosed definitely according to the clinical symptom, and the examination with electromyogram is beneficial to diagnose this disease, but it has not specificity. Most patients of supinator syndrome often get satisfactory effects after conservation treatment, and the ones who are failed can get the improvement from surgical treatment timely.
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@#Objective To analyze the diagnosis and treatment of cervical spinal cord injury without fracture or dislocation.Methods 38 patients were analyzed retrospectively.Results and Conclusion According to the manifestation of MRI and the symptoms,6 cases accepted conservation treatment,and the other 32 cases accepted surgery with different styles.All the patients recovered in some degrees.
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This paper analyzed the effects of prosthesis selection and surgical technique in total knee arthroplasty (TKA) on patellofemoral joint biomechanics, patella movement track and prosthesis rotational alignment to explore the mechanics-related factors of knee joint function and prosthesis service life after TKA. Traditional anatomic landmark line lacks of accuracy due to various factors, so there is no uniform linear localization for tibial prosthesis. Femoral epicondylar axis functions as the reference axis of prosthesis rotational alignment in TKA, which can achieve ideal patellofemoral joint biomechanics, and reduce postoperative complications. 1/3 line of medial border to moderate medial border of the tibial tubercle is regarded as the ideal rotational interval o the tibial prosthesis, and introversion and extroversion degrees of the knee joint are determinant of linear localization. Knee symptoms and function are significantly improved after fixed and mobile-bearing arthroplasties. These two kinds of prosthesis do no significantly vary in knee joint, pain, function, and patella scores as well as radiography. However, the knee motions during maximum flexion in the mobile-bearing arthroplasty are smaller than in the fixed one. It has been proven by experimental studie that mobile-bearing arthroplasty shows wearing resistance and joint movement superior over the fixed one, but there is no agreemen in clinical studies.