0.05).The median therapy fees in the open group was nine thousand yuan vs.eleven thousand yuan in the laparoscopic group,and the difference was significant(P

BackgroundPosterior capsule opacification(PCO) is the main cause inducing low vision after extacapsular cataract extraction. Our previous study determined that polylysine-ethylene diamine tetraacetic acid (EDTA) (PLE) can suppress the incidence of PCO. ObjectiveThe goal of this experiment was to investigate the inhibition of polylysine-EDTA on rabbit lens epithelial cells (LECs)proliferation in vitro and the effective concentrations of polylysine-EDTA. MethodsThe anterior capsular membranes from 10 3-month-old clean New Zealand white rabbits were digested and then cultured to obtain the LECs. The second and third generation of LECs were inoculated on the 96-hole culture plate with the cell density of the 1 × 105/ml. 12.5,25.0,50. 0,100. 0 μmol/Lof PLE were added into the culture medium for 48 hours respectively,and the DMSO medium was used at the same way as the control group. The proliferation of the LECs was then detected by MTT method and the inhibitory rate of PLE on LECs growth was calculated. ResultsLECs grew at a near normal state in ≤25.0 μmol/L PLE groups,however,cultured LECs were out of shape and the numbers decreased with the weakened adhesion ability in ≥50.0 μ mol/L PLE groups. The A490 values of LECs were 0. 278±0. 013,0. 266±0. 028,0. 260±0. 022 and 0. 247±0. 012 in 12. 5,25.0,50. 0, 100. 0 μmol/L polylysine-EDTA groups respectively and were lower than 0. 311 ±0. 038 of DMSO control group( P=0. 035,0. 011,0. 009,0.013 ). The inhibitory rates of 12. 5,25.0,50. 0, 100.0 μmoL/L PLE on LECs proliferation were 10.61％ , 14.47％ , 16.40％ and 20. 58％ respectively. ConclusionsPolylysine-EDTA can inhibit the growth and proliferation of LECs in vitro at a dose-dependent manner.

Objective To investigate the feasibility and clinical effect of cannulated screws plus separate vertical wirings technique for acute fracture of the inferior pole of the patella.Methods From May 2012 to September 2013,14 patients with fresh closed unilateral fracture of the inferior pole of the patella were treated with the cannulated screws plus separate vertical wirings.Eight patients were injured in traffic collisions and 6 in fall accidents.Fracture AO classification was type 34A1 in 8 patients and type 34A2 in 6 patients.Time from injury to operation was 1-7 days (mean,2.5 days).Number of tie wires was determined according to the degree of fracture comminution.Fracture healing,fixed position and patellar length were evaluated by radiographic examination postoperatively.Knee mobility and Bostman evaluation system were investigated to analyze the clinical effect.Results All the patients obtained average 15-month follow-up (range,12 to 29 months).At postoperative 2 months,the fracture healed with good alignment of the broken bone and proper place of the internal fixation device noted on the X-ray films.At postoperative 6 and 12 months,X-ray films revealed fracture bony healing,good location of the wire internal fixation,and no apparent shortening of the patella.At the 12 months,range of knee motion was (126.0 ± 4.5) ° for flexion and (2.0 ± 1.7) ° for extension.Bostman functional score for patella fracture was (28.1 ± 1.9) points.And 12 patients were rated as excellent and 2 good,with excellence rate of 100％.Conclusion Cannulated screw fixation plus separate vertical wiring is effective to stabilize patella inferior pole fracture and has good results,indicating a recommended surgical method.

Objective To investigate the clinical features,karyotype,and the prenatal diagnosis for his sibling of a Chinese patient with rare ring chromosome 20 syndrome induced intractable epilepsy.Methods The clinical data of the patient diagnosed in Peking University People's Hospital were collected.The clinical manifestations,chromosome karyotype were summarized.Results The proband,a boy,started to show intermittent tonic seizures or atypical absence seizures and psychomotor retardation from the age of 11 months.Several anti-epilepsy drugs and globulin had been tried without effect.Common karyotype analysis and epilepsy-related genes analysis revealed no abnormality.However,abnormal karyotype 46,XY,r(20)(p13q13.3) in his peripheral blood lymphocytes was found by high resolution chromosome karyotype analysis with 550 G-banding,and the diagnosis of ring chromosome 20 syndrome,type Ⅱ was confirmed.The mother of the patient underwent amniocentesis at the midterm of the second pregnancy.The cultured amniocytes karyotypes were normal.The second child(a boy) of the family was 1 year old without epilepsy and the psychomotor development was normal.Conclusions Ring chromosome 20 syndrome is a rare human chromosome abnormality.The syndrome is associated with epileptic seizures,behavior disorders and mental retardation.Since karyotype testing is not a routine investigation for the patient with epilepsy,the diagnosis of ring chromosome 20 syndrome is usually delayed or misdiagnosed.The karyotype analysis should be considered for the etiological study of the patients with intractable epilepsy with unknown origin.

OBJECTIVETo establish a long-term proliferation culture system for mouse spermatogonial stem cells.

METHODSTestis tissues were obtained from 30 newborn male ICR mice on postnatal day 2-6. Testis cell suspension was collected by two-step enzymatic digestion prior to culture. The dissociated cells were aliquoted into tissue culture plates and cultivated with a modified system composed of serum-free defined medium on mouse embryonic fibroblasts (MEF) feeders. Their proliferation was determined by the BrdU incorporation test and the cultured cells identified by alkaline phosphatase (AP) activity, immunofluorescence staining and RT-PCR assay.

RESULTSThe cultures remained in a steady state and continued to generate germ cell colonies. The undifferentiated state was confirmed by strong positivity for AP activity, immunofluorescent staining of GFRalpha-1+ /Oct-4+ /VASA+ /SCP3- and GFRalpha-1+ /Oct-4+/SCP3- at the gene expression levels.

CONCLUSIONMouse spermatogonial stem cells could be expanded in our defined culture system and passaged steadily in vitro. The harvested cells remained in an undifferentiated state, which has provided a good platform for the study of spermatogenesis in vitro.

Alkaline Phosphatase ; genetics ; metabolism ; Animals ; Cell Culture Techniques ; Cell Proliferation ; Cells, Cultured ; Fluorescent Antibody Technique ; Male ; Mice ; Mice, Inbred ICR ; Reverse Transcriptase Polymerase Chain Reaction ; Spermatogonia ; cytology ; metabolism ; Stem Cells ; cytology ; metabolism ; Time Factors

objective To compare three risk prediction system,the physiological and operative severity score for the enumeration of mortality and morbidity(POSSUM),the Portsmouth POSSUM (P-POSSUM)and the colorectal POSSUM(Cr-POSSUM)for the accuracy in predicting operative mortality of patients of colorectal cancer in a single Chinese referral hospital setting. Methods Data of 903 patients,who undergone surgery for colon and rectal cancers from 1992 to 2005 at Peking University Third Hospital,were enrolled in the study.POSSUM,P-POSSUM and Cr-POSSUM was used respectively to predict the mortality rate.ROC curve was applied to judge the differentiation ability of each score.Model goodness-or-fit was tested by the Hosmer-Lemeshow statistic and subgroup analysis was performed by the ratio of observed to expected deaths(O∶E ratio). Results The actual inhospital mortality in our series was 1.0%(9/903).The oredicted mortality rate by POSSUM,P-POSSUM and Cr-POSSUM were 5.6%,2.8% and 4.8%respectively.These predicted mortality rate were significantly higher than actual mortality of our patients.The O∶E ratio was 0.18,0.35 and 0.2 respectively. Conclusion The predicted mortality rate of POSSUM,P-POSSUM and Cr-POSSUM were significantly higher than actual observed mortality rate in a single Chinese referral hospital for patients of colorectal cancer.

Objective To evaluate a score system(Association of Coloproctology of Great Britain and Ireland ACPGBI)in prediction of postoperative mortality from colorectal cancer patients in a Chinese hospital. Methods We analyzed retrospectively 904 patients with histologically confirmed colorectal cancer who had colorectal surgery from 1992 to 2005.There were 525 colonic cancer patients and 379 rectal cancer patients.We divided patients into several groups according to operative urgency(elective or emergency);surgeons(colorectal specialists or other surgeons);cancer location(colon or rectal).According to ACPGBI score we got the prediction.This prediction was compared with the actual mortality;Chi-square test,receiver operator characteristic curve(ROC),Hosmer-Lemeshow goodness-of-fit test were used.Results Observed overall mortality within 30 days after surgery was 1.0%(9/904),and the predicted mortality was 8.3%(75/904).In all the subgroups the predicted momdity wag higher than observed mortality.We found that the actual mortality was higher in an individual subgroup in which the predicted mortality was higher. Conclusions For colorectal cancer patients undergoing a surgery the predicted mortality of ACPGBI score system was higher than the actual mortality in a Chinese hospital.

A strain of grass carp reovirus was isolated from sick grass carp with symptoms of haemorrhage in Guangdong province in 2009. The strain was tentatively named as GCRV-GD108 because it was isolated from grass carp and possessed 11 segments of dsRNA. Complete genome sequence analysis showed that significant differences existed between GCRV-GD108 and GCRV, as well as other known species of aquareovirus. In this study, molecular characteristics of diseased grass carp collected from different farms in Guangdong, Fujian and Hunan provinces were assayed. Based on the sequences of the 11 segments of GCRV-GD108, PCR primers corresponding to each of the segments were designed and synthesized. Total RNA of the diseased fish was extracted and used as templates of RT-PCR reaction. Specific amplification bands were obtained from all of the samples whereas no band was produced from GCRV standard strain. While using the primers specific to GCRV produced specific band in GCRV standard strain rather than in these collected samples. Sequencing of the amplification products showed that these samples displayed high similarities with each other (95.2%-99.4%), and they also shared high sequence similarities with that of GCRV-GD108 (95.0%-99.8%), suggesting that these samples shared similar molecular characteristics with those of GCRV-GD108, and were quite different from GCRV as well as the known species of genus aquareovirus. The results indicated that there are different molecular types of reovirus existed in the pond-cultured grass carp in China, and GCRV-GD108 is a representative strain in southern China, therefore great attention should be paid in order to control the disease efficiently, especially in vaccine preparation. Two pairs of primers were chosen to establish a duplex PCR assay method by combining each pair of the primers specific to GCRV-GD108 with the GCRV primer pair respectively. The duplex PCR assay method will enable the identification of GCRV-GD108 or GCRV by only a single PCR reaction.
Animals ; Carps ; virology ; China ; Fish Diseases ; diagnosis ; virology ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Reoviridae ; genetics ; isolation & purification ; Reoviridae Infections ; veterinary ; virology

Abstract: Objective　To develop a real-time fluorescent quantitative RT-PCR (qRT-PCR) method for qualitative and quantitative Chikungunya virus (CHIKV) analysis. Methods Based on the systematic analysis of the genomic sequences of Chikungunya and its related arboviruses, the specific nucleic acid sequences for Chikungunya virus were screened and identified, and then the primers and TaqMan probe were designed. Meanwhile, the human GAPDH gene was used as an internal reference. The reaction system for qRT-PCR was systematically optimized by L9(34) orthogonal design, and a rapid detection method for Chikungunya by qRT-PCR based on TaqMan probe methods was established. The sensitivity, specificity, reproducibility, and coverage of the established method were analyzed in detail. The standard curve was made, and the absolute quantitative method was established using the cloned nucleic acid fragments as positive samples. Results　A real-time fluorescent quantitative RT-PCR assay was developed for the qualitative and quantitative analysis of Chikungunya virus. The reaction system included Chikungunya virus and reference internal gene specific primers and probe, RT/Taq enzyme mixture, reaction buffer, and negative and positive reference. The established method obtained positive results with the ROSS strain of ECSA subtype, LR2006 strain of IOL branch, 181/25 strain of Asian type and Dongguan 2010 epidemic strains of Chikungunya virus, but there was no cross-reaction with other 18 arboviruses belonging to Flaviviruses, Alphaviruses and Bunyavirus. The minimum detection limit of the established method was 5.80 copies/mL, and a linear relationship was observed between the amount of input plasmid DNA and fluorescence signal value over a range of 5.80×102 copies/mL to 5.80×1010 copies/mL, and the correlation coefficient was 0.999 5. The qRT-PCR amplification efficiency was 91%, and the intra-assay variations and inter-assay variations were 0.01-0.07 and 0.03-0.11, respectively. Conclusions　The TaqMan qRT-PCR method developed in this study can qualitatively and quantitatively detect Chikungunya virus rapidly with specificity and sensitivity, providing a technical method for the prevention and control of this viral disease.

OBJECTIVETo analyze the genetic polymorphism of 5 short tandem repeat(STR) loci in Hui population in Ningxia area.

METHODSThe genetic polymorphisms of five selected STR loci(D11S1984, D14S306, D14S617, D17S1290 and D19S433) in chromosomes 11, 14, 17 and 19 in 144 unrelated individuals in Hui population in Ningxia area were analyzed by PCR amplification, denaturing polyacrylamide gel electrophoresis(PAGE) and silver staining.

RESULTS10, 8, 11, 13 and 8 alleles, 30, 25, 33, 40 and 23 genotypes of the 5 STR loci in Hui population in Ningxia were detected. The measured values of the heterozygosity of the 5 STR loci were 0.8413, 0.8033, 0.8331, 0.8369 and 0.7703; of the polymorphism information content were 0.8217, 0.7746, 0.8121, 0.8174 and 0.7332; of the discrimination power (DP) were 0.9516, 0.9257, 0.9611, 0.9660 and 0.9135. The calculated discrimination power was 0.9999995. The measured values of paternity exclusion were 0.7046, 0.6367, 0.6911, 0.7012 and 0.5801; the calculated paternity exclusion was 0.9958. The genotype distributions were in accordance with Hardy-Weinberg equilibrium.

CONCLUSIONThe 5 STR loci have better polymorphism in Hui population in the Ningxia area, and thus could serve as useful markers for population genetics research and for individual identification and paternity test in forensic medicine.

Asian Continental Ancestry Group ; genetics ; China ; Female ; Gene Frequency ; Genotype ; Heterozygote ; Humans ; Male ; Microsatellite Repeats ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic