Microcell mediated chromosome transfer (MMCT) is a challenging technique for introducing exogenous chromosomes into interested mammalian cells. Combined with the somatic cell nuclear transfer technique, MMCT has been employed for producing transchromosomic animals of medical and agricultural value. Producing high quality of microcells is a key step in the success of MMCT. Eamamined by fluorescin staining and Giemsa staining, 0.2 mg/L colcemid was considered suitable for inducing high percentage of micronuclei in A9 (neo12) cells, without causing death of a mass of cells. Microcells were produced by centrifugation of micronucleated A9 (neo12) cells in Percoll density gradient containing 20 mg/L Cytochalasin B at 39 000 g. The resulting mixture of microcells, whole cells, karyoplasts and cytoplast fragments was filtered through 8 μm and 5 μm size membrane pores sequentially to obtain pure preparation of microcells. Microcells were then characterized by Giemsa staining and microcell PCR was first applied for examination of the quality of microcell preparation. The result showed that microcells containing our interest chromosomes-human chromosome 12 were equally distributed in the preparation, the preparation was suitable for use in generation of transchromosomic animals.

Stroke is one of the most devastating complications after cardiac surgery,and contributes to both mortality and morbidity.Coronary artery bypass graft (CABG) and percutaneous coronary intervention (PCI) are the common procedures for the treatment of coronary artery disease.This article reviews the pathophysiologic mechanisms,risk factors,treatment and prognosis of stroke after CABG and PCI.

Objective To investigate the correlation of serum interleukin-6 (IL-6) and C-reactive protien (CRP) lev?els with left atrial thrombus and severe spontaneous echocontrast (SEC) in patients with atrial fibrillation (AF). Methods A case-control study of patients with atrial fibrillation (n=76) was carried out. All patients were divided into control group (n=45) and study group (n=31) according to their conventional echocardiography performance. Serum IL-6 and CRP were exam?ined in both groups. Results The levels of serum IL-6 in patients with thrombus and severe SEC were (324.13±42.86) ng/L and (332.29±53.17) ng/L, respectivly which is higher than that in patient without thrombus or without severe SEC (108.75± 25.43) ng/L and (93.59 ± 27.82) ng/L respectively. In parallel, CRP levels in patients of thrombus and severe SEC were (66.97 ± 17.65) mg/L and (71.81 ± 20.19) mg/L respectively which is higher than that in patients without thrombus or without severe SEC (17.28±6.52) mg/L and (16.76±8.73) mg/L respectively. All differences were of statistically significant(P<0.05). Conclusion Increase of serum IL-6 and CRP as well as high systemic inflammatory state correlate with left atrial thombus and severe SEC in patients with AF.

ObjectiveTo evaluate the value of the measurement with a scale in arthroscopic anatomic anterior cruciate ligament (ACL) reconstruction.Methods Thirty-six patients with unilateral ACL-deficient knees were divided into single bundle group and double bundle group based on the length of the major axis of the native ACL footprint measured with a scale.Consistent with the International Knee Documentation Committee (IKDC) scoring systems,Lysholm knee scoring scale and Tegner activity rating were used to evaluate the clinical results,and data from 23 to 26 months follow-up were gathered and analyzed statistically.ResultsIn terms of IKDC evaluation,19 patients( 90.4％ ) in the double band group and 13 patients( 86.7％ )in the single band group were graded as normal or nearly normal ( Z =7.82,6.33 ; P ＜ 0.05 ).The mean Lysholm scores were 93.8 ± 9.9 and 94.2 ± 9.6 and the mean Tegner scores were 6.6 ± 1.9 and 6.5 ± 1.7 for the double and single band groups,respectively( t =2.31,5.60,3.55,7.09 ;P ＜0.05 ).ConclusionArthroscopic measurement with a scale combined with other traditional methods to determine the tunnel location is easy to conduct and intuitional to understand.It could be useful in anatomic ACL reconstruction.

Purpose　The aim is to isolate and purify the erythropoietin(EPO)and thrombomodulin(TM) from human urine.Methods　Purifying and isolating human urine with means of filtration and concentration,ion-exchange chromatography, affinity chromatography and so on.Results　4.47 mg EPO and 9.92 mg TM were obtained from 2 000 kg human urine.Their molecular weights were(38.6±1.0)kD and (60±1.4)kD erespectively, the isoelectric point, 3.60±0.02 and 3.70±0.02, yield, 31.9% and 25.0%.The purified EPO and TM was shown to be highly homogeneous by capillary zone-electrophoresis(CZE),abundant Glu，Ala and Gly include on TM. Conclusion　The high purity EPO and TM was obtained.

OBJECTIVE:To establish a method for the determination of related substances in bisacodyl raw material and enteric-coated tablet. METHODS:HPLC was performed on the column of Hibar C18 with mobile phase of acetonitrile-20 mmol/L ammonium acetate (acetic acid adjust pH to 5.0)(55∶45,V/V),detection wavelength was 265 nm,flow rate was 1.0 ml/min, column temperature was 30℃,and the injection volume was 20 μl. RESULTS:The linear range of bisacodyl was 0.25-5.0 mg/ml (r=0.999 9);the limits of detection and quantification were 19-25 ng and 61-68 ng for bisacodyl and impurity A,B,C,D and E;RSDs of precision,stability and reproducibility tests were lower than 2%;recovery was 99.50%-101.00%(RSD=0.5%,n=9). CONCLUSIONS:The method is specific, sensitive and reproducible, and can be used for the determination of related substance in bisacodyl raw material and enteric-coated tablet.

Objective To explore the differences in clinical and pathological characteristics of idiopathic membranous nephropathy between elderly and non elderly patients.Methods The clinical and pathological data,which were used for biopsy diagnosing idiopathic membranous nephropathy in elderly patients (aged ≥65 years) versus non elderly patients (aged ＜65 years) in our hospital during the recently 10 years,were retrospectively analyzed and compared.Results The elderly patients with idiopathic membranous nephropathy accounted for 26.29％ and patients aged less than 45 years with idiopathic membranous nephropathy accounted for 25.26％.The proportions of idiopathic membranous nephropathy in the two different age groups were similar.The occurrence rate of hypertension and microscopic hematuria as well as levels of blood serum creatinine and urinary N-acetyl-β-D-glucosaminidase were higher,while the urinary osmotic pressure,estimated glomerular filtration rate (eGFR) and blood albumin were lower in elderly group than in non-elderly group (all P＜0.05).The proportions of glomerulosclerosis,tubulointerstitial lesions and vascular lesions were higher in elderly group than in non-elderly patients (all P＜ 0.05).There were no significant differences in the immunofluorescence intensity of IgG,positive rates of IgA,C3,C4 and Fi between the two groups,while the positive rate of C1q was lower in elderly group than in non elderly.Among 51 patients receiving IgG4 immunohistochemical staining,the positive rate of IgG4 staining and the intensity were higher in elderly group than in non-elderly [100.00％ vs.81.25％,(2.11±0.46) vs.(1.59 ± 0.95),both P ＜ 0.05].Conclusions The proportion of idiopathic membranous nephropathy is increased in non-elderly patients.Elderly patients are more susceptible to hypertension,hematuria and renal dysfunction.The positive rate and the intensity of IgG4 are higher and the positive rate of C1q is lower in elderly group than in non-elderly group,which indicate that patients with negative IgG4 and positive C1q have secondary causes which have not yet been demonstrated,and they should be closely observed during the follow-up.

Objective To compare the effect of ISO-C3D navigation and conventional C-arm fluoroscopy in iliosacral screws insertion for treatment of posterior pelvic ring injuries.Methods Sixty-five patients with posterior pelvic ring injuries managed by minimally invasive percutaneous iliosacral screws from June 2006 to January 2012 were reviewed.There were 37 males and 28 females,at age range of 18-63 years (mean 35.9 years).Pelvic fracture classification based on Tile system was type B1 in 10 cases,type B2 in 15,type B3 in nine,type C1 in 18 and type C2 in 13.Patients were divided into ISO-C3D navigation group (Group A,n =35) and C-arm fluoroscopy group (Group B,n =30) according to the difference in intraoperative fluoroscopy methods.Intraoperative fluoroscopy time,time cost in inserting a screw,patient satisfaction rate for bone reduction,bone union time and excellent-good rate of postoperative function were recorded.Results Eighty cannulated screws were inserted for the 65 patients.Average fluoroscopy time and time cost in inserting a screw were shorter in Group A than in Group B (P ＜0.01),but there was no statistical difference between the two gronps in patient satisfaction rate for bone reduction.No patient presented with infection,vascular nerve injury or other complications.Follow-up was 6-24 months (mean 12.7 months) for all the patients.Functional recovery showed no statistical difference between the two groups at postoperative 6 months.All fractures were healed and no delayed union or nonunion happened.Conclusion As compared with conventional C-arm fluoroscopy,computer-navigated surgery can reduce fluoroscopy time and improve screw insertion accuracy.

Objective To understand the enterovirus infection in family close contacts of patients with hand,foot and mouth disease (HFMD) and the contamination of environment.Methods Forty-one HFMD cases confirmed by laboratory from web-based surveillance system during July to August 2010 in Guangdong Province were selected.All members of the cases′ family were investigated by collecting their information on demography,habit of domestic hygiene and hygiene status in household.The stool samples of all members and the smear samples from the surface of family belongings from 16 families were collected and the enterovirus was detected by real time quantitative polymerase chain reaction.The data were analyzed by chi square teat and t test.ResultsForty-one HFMD cases′ families and 135 close contacts were included in this survey.The infection rate of the enterovirus was 39.2％ (53/135) in all close contacts.Of all the investigated families,the infectionrate was 58.5％ (24/41) in family with one or more close contacts and 9.8％ (4/41) in family with all close contacts.The differences of infection rates of enterovirus among the members of parents (32.5％,25/77),grandparents/aunts/ uncles (43.3％,13/30) and cousins (53.6％,15/28) didn′t show statistical significance (χ2 =4.07,P=0.131).The infection rate of enterovirus in close contacts from family with more than 5 members was higher than that from family with 4 or less members (OR=1.4,95％CI 1.1-1.9).Among 135 close contacts,27.4％ (37/135) were infected with the same types of entervirus as that of HFMD case in the family and 11.9％ (16/135) were infected with the different virus types.In 33 family belongings samples from 16 families,the positive rate of enterovirus detection was 6.1％ (2/33).Between 17 families with enterovirus testing negative and 23 families with enterovirus testing positive in close contacts,there were no statistical differences of the family hygiene status,hand-washing of babysitter,disinfection of tableware and drinking,sharing towels,airing bedding articles and toy cleaning (P＞0.05).ConclusionsThe infection rate of enterovirus in close contacts of HFMD cases is high and the enterovirus contamination exists in case family environment.Management of close contacts of HFMD cases and disinfection of the family environment are important in HFMD controls.

Objective:To investigate the effects of aerobic combined stretching exercise during dialysis on the severity of restless legs syndrome(RLS), quality of life and sleep quality in hemodialysis (HD) patients.Methods:A total of 63 HD patients with RLS who were admitted to the Hemodialysis Center, Renmin Hospital of Wuhan University from March 2018 to March 2019 were selected as research objects. According to the random number table method, they were divided into the observation group (32 cases) and the control group (31 cases). Patients in the control group were given conventional nursing methods while patients in the observation group were given the aerobic combined stretching exercise during dialysis. The International Restless Legs Syndrome Rating Scale (IRLS) Kidney Disease Quality of Life, (KDQOL-36 TM), Pittsburgh Sleep Quality Index(PSQI) and blood chemistry were used to evaluate the effect of exercise on HD patients with RLS. Results:After intervention, the observation group ′s IRLS scores was 12.19±4.48, lower than the control group (17.87±3.91), the difference was statistically significant ( t value was 3.021, P<0.05). The scores of physical health, mental health, kidney burden, symptoms and discomfort, effects of kidney disease in KDQOL-36 TM of the observation group were 44.56±6.64, 46.12±4.95, 19.92±15.91, 87.59±5.65, 64.55±13.20, higher than the control group (37.43±4.81, 41.81±4.87,12.10±14.87, 78.83±7.96, 55.45±15.52), the differences were statistically significant ( t values were -5.027--2.015, P<0.05). The PSQI total score was 8.94±2.54, lower than that of the control group (13.13±1.31), the difference was statistically significant ( t value was 8.271, P<0.05). The blood phosphorus was (1.80±0.48) mmol/L in the observation group, lower than (2.04±0.34) mmol/L in the control group, and hemoglobin, blood calcium [(105.31±13.58) g/L, (2.26±0.20) mg/L] were higher than those in the control group[(99.52±8.21) g/L, (2.05±0.29) mg/L], the differences were statistically significant ( t values were 2.275, -2.042, -3.325, P<0.05). Conclusions:The aerobic combined stretching exercise during dialysis relieves the symptom of RLS, improves the sleeping quality and quality of life.