Objective To evaluate the clinical effect of tibial plateau bone defects repair using osteotomy and bone grafting with autogenous bone gain way in total knee arthroplasty.Methods From Mar.2011 to Mar.2015,28 knees of 20 patients with bone defects in medial tibial plateau were in total knee arthroplasty,the bone defects were repaired into horizontal steps,then with autogenous bone grafting by osteotomy.Results The bone defects could be repaired effectively to support the knee joint prosthesis,all the patients were followed-up through outpatient from 1 year to 4 years with the average of 2.6 years,the HSS kness score increased from (20.9 ± 1.2) before opearation to (87.6 ± 1.6) after operation.Hss score were paired t test,there was statistically significant difference (P ＜ 0.05).The location was good and there was no bone defect image,no bone resorption and no joint dislocation under X-ray.Conclusions To repair the bone defects in total nkee arthroplasty,it can use the autogenous bone by osteotomy reasonably,achieve good repair defect site,reserve the bone mass maxium,reconstrust the balanced mechanical support of tibia.

Objective To formulate a five-level pediatric emergency triage standard and evaluate the efficacy and reliability of it in determining severity of emergency pediatric patients.Methods According to the assessment methods in Pediatric Advanced Life Support recommended by American Heart Association and American Academy of Pediatrics,we formulated a five-level pediatric emergency triage standard based on the situation of our hospital and analyzed the data one year before and after the application of it.Results Before and after the application of the triage standard,the average satisfaction rate of emergency patients were (81.28 ± 3.97) ％ and (94.13 ± 4.62) ％,and there was significant difference (P ＜ 0.01) ; the proportion of whom became worse during waiting time were 1.83％ (628/34275) and 0.04％ (16/36 187),and there was significant difference (P ＜0.01) ;the average waiting time of emergency admission patients were (12.71 ± 2.32) min and (3.34 ± 1.95) min,and there was significant difference (P ＜0.01) ;the misjudgment rate of severity were 3.78％ (1 296/34 275) and 0.57％ (205/36 187),and there was significant difference (P ＜ 0.05).Conclusion The five-level triage standard is objective,easy to master,and suitable for pediatric triage.It can quickly sort out critical cases from emergency pediatric patients,which can improve the effectiveness of emergency service,make use of medical resources rationally and somewhat solve the problem of overcrowding.

Anticonvulsants ; adverse effects ; Humans ; Seizures ; chemically induced

Background Laser peripheral iridoplasty (LPI) is widely used in the treatment of glaucoma by flattening the iris and widening angle of anterior chamber (AA).However,no evidence suggests the optimal site of LPI in iris.Objective This study was to compare the therapeutic effects of LPI at different sites of iris for glaucoma.Methods Glaucoma models were established in the right eyes of 40 healthy adult male pigment rabbits by intrachamber injection of 0.1 ml compound carbomer solution with 0.3％ carbomer and 0.025％ dexamethasone.The models were randomly divided into model control group,corneoscleral limbus group,one spot from corneoscleral limbus group and two spots from corneoscleral limbus group.LPI was performed at corresponding site of iris by 532 nm argon laser with the spot diameter 500 μm,energy 300 mW,exposure time 0.3 seconds and laser number 24 spots,and the rabbits in the model control group did not receive LPI.Intraocular pressure (IOP),coefficient of outflow facility (C value) were measured and calculated with Schi(o)tz tonometer before LPI and 2,4,7,14 and 30 days after LPI,and anterior chamber depth (ACD),AA,anterior chamber angle opening distance within 500 μm radius from scleral spur (AOD500) were measured with ultrasound biomicroscope (UBM).The eyeballs were extracted 30 days after LPI,and the chamber angle were observed under the optical microscope after hematoxylin and eosin staining.The use and care of the animals complied with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health.Results UBM showed that compared with the model control group,the anterior chamber angle was evidently widened in all the LPI groups,with the best effectiveness in the one spot from corneoscleral limbus group and the worst one in the two spots from corneoscleral limbus group.Compared with the model control group,the IOP was evidently reduced,and C values,AA and AOD500 were significantly increased in the corneoscleral limbus group,one spot from corneoscleral limbus group and two spots from corneoscleral limbus group after LPI,showing significant differences among the four groups (IOP:Fgroup =16.848,P ＜ 0.01;C value:Fgroup =9.629,P ＜ 0.01;AA:Fgroup =62.336,P＜0.01;AOD500:Fgroup =77.779,P ＜ 0.01).IOP was reduced and C value,AA and AOD500 were increased in 2,4,7,14 and 30 days after LPI as compared with before LPI,with significant differences over time (IOP:Ftime =3.041,P =0.011;C value:Ftime =4.311,P＜0.01;AA:Ftime =14.627,P＜0.01;AOD500:Ftime =20.378,P＜0.01).Compared with the model control group,the ACD was significantly increased in the corneoscleral limbus group and one spot from corneoscleral limbus group,and that in the two spots from corneoscleral limbus group was significantly reduced,and the ACD was insignificantly increased over time after LPI (Fgroup =18.017,P＜0.01;Ftime =0.022,P =1.000).Hematoxylin and eosin staining showed that the trabecular meshwork and adhesion of tissure were reopened and the anterior chamber angle was widened after LPI.Conclusions LPI can widen anterior chamber angle and lower the IOP.The best therapeutic outcome for glaucoma is displayed when LPI is performed at the iris site corresponding to one spot from the corneoscleral limbus.

OBJECTIVETo explore the mechanism of Alpha-fetoprotein (AFP) effects on hepatocellular carcinoma cells (HCC) resistances apoptosis induced by tumor necrosis factor-related apoptosis inducing-ligand (TRAIL).

METHODSThe expressed alteration of TRAIL receptor-2 (DR5) after the human hepatoma cells line Bel 7402 (AFP-producing) and HLE cells (non-AFP producing) were treated with all trans retinoic acid (ATRA) were determined by Western blot; Interaction of AFP with RAR-beta was analyzed by co-immunoprecipitation (Co-IP); Laser confocal microscopy was used to observe co-localization of AFP and RAR-beta; Short small RNA interfering (RNAi) was applied to knock down the expression of AFP in Bel 7402 cells; The full AFP gene cDNA was inserted into pcDNA3.1 vector and constructed the expressed vector of AFP (named pcDNA3.1-afp); The growth of hepatoma cells was analyzed by MTT.

RESULTSBel 7402 and HLE cells expressed DR5, lowed dosage of ATRA (40mumol/L) had no influence on the expression of DR5 in Bel 7402 cells, but ATRA (160mumol/L) could inhibit the expression of AFP and promote the expression of DR5 significantly; Co-IP indicated that AFP had a property for interacting with RAR-beta; The results also demonstrated AFP co-localization with RAR-beta in cytoplasm of Bel 7202 cells; The expression of DR5 was enhanced while the expression of AFP was knocked down by RNAi. pcDNA3.1-afp vector was transfected into HLE cells, the growth of HLE cells were stimulated and TRAIL cytotoxicity of HLE cells were reduced. But when the expression of AFP was knocked down the sensitivity of Bel 7402 cells to TRAIL was enhanced.

CONCLUSIONSThese data provided that AFP had a capability to interact with RAR-beta and suppressed the expression of DR5. AFP could play pivotal role on hepatoma cells resistance-induced apoptosis by TRAIL.

Apoptosis ; Cell Line, Tumor ; metabolism ; Humans ; Receptors, Retinoic Acid ; metabolism ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; metabolism ; TNF-Related Apoptosis-Inducing Ligand ; metabolism ; Tretinoin ; pharmacology ; alpha-Fetoproteins ; metabolism

OBJECTIVETo investigate single nucleotide polymorphisms (SNPs) and the distribution of their haplotypes in caspase-8, -10 genes in Zhejiang Han nationality in China.

METHODSPCR, denaturing high-performance liquid chromatography (DHPLC) and DNA sequencing were used to detect the SNPs in the 2nd-5th exons of caspase-10 gene, the 8th-10th exons of caspase-8 and their flanking sequences. Expectation Maximization (EM) algorithm was used for haplotype frequencies analysis and pairwise linkage disequilibrium (LD) test.

RESULTS(1) Two SNPs, A2823G and A12799G, were identified in caspase-10 gene, located in exon 2 and exon 5 respectively. A12799G was newly found with low informativeness. Three SNPs were identified in caspase-8 gene; A43466G, G51484A and G52951A were located in exon 8, exon 9 and intron 9, respectively. They do not change the primary structure of the encoded protein. (2) Linkage equilibrium was observed between A2823G in caspase-10 gene and the three sites in caspase-8 gene. A43466G and G52951A, and G51484A and G52951A in caspase-8 gene were also in linkage equilibrium. Their coefficients of disequilibrium were near 0. Whereas strong linkage disequilibrium was observed between A43466G and G51484A, because its coefficient of disequilibrium was near 1. (3) A total of 11 haplotypes were estimated within A2823G in caspase-10 gene and three sites in caspase-8 gene. A-2823/A-43466/G-51484/G-52951 was the main haplotype with a frequency of 0.3811. A-2823/A-43466/G-51484/A-52951 was the second haplotype with a frequency of 0.2536. The polymorphism information content of their haplotypes was 0.7106.

CONCLUSIONThe SNPs of caspase-8, -10 genes in Han Chinese of Zhejiang could be parsed into at least three different haplotype blocks. The polymorphism information content can be improved by using haplotype analysis of several SNPs.

Alleles ; Base Sequence ; Caspase 10 ; genetics ; Caspase 8 ; genetics ; China ; ethnology ; DNA ; analysis ; Ethnic Groups ; genetics ; Gene Frequency ; Haplotypes ; Humans ; Polymorphism, Single Nucleotide

OBJECTIVETo design and synthesize ribozymes targeting 138 and 218 sites of the mRNA nucleotide of mouse caspase-12, a key intermedium of ER stress mediated apoptosis, and to identify their activities through in vitro transcription and cleavage.

METHODSThe mouse caspase-12 gene fragment was obtained by RT-PCR and cloned into the PGEM-T vector under the control of T7 RNA polymerase promoter. The transcription product of the target was labeled with a-32P UTP, while ribozymes were not labeled. Ribozyme and target RNA were incubated for 90 min at 37 degree C in a reaction buffer to perform the cleavage reaction.

RESULTSIt was found that under a condition of 37 degree C, pH 7.5 and with Mg2+ in a concentration of 10 mmol/L, Rz138 and Rz218 both cleaved targets at predicted sites, and the cleavage efficiency of Rz138 was 100%.

CONCLUSIONRz138 and Rz218 prepared in vitro possess the perfect specific catalytic cleavage activity. Rz138 has excellent cleavage efficiency. It may be a promising tool to prevent ER stress induced apoptosis through catalytic cleavage of caspase-12 mRNA in vivo. It also can be used to verify whether caspase-12 is necessary in ER stress induced apoptosis.

Animals ; Base Sequence ; Caspase 12 ; genetics ; Endoplasmic Reticulum ; metabolism ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Oxidative Stress ; genetics ; RNA, Catalytic ; chemistry ; genetics ; RNA, Messenger ; genetics

OBJECTIVETo design hammerhead ribozymes against mouse caspase-7 and to study their expression and cleavage activity in vitro.

METHODSThe secondary structures of ribozyme and caspase-7 genes were analyzed and simulated by computer. Ribozymes DNA sequences were synthesized by automatic synthetic apparatus. Caspase-7 DNA sequence was acquired by reverse transcription PCR. Ribozymes and caspase-7 DNA sequences were separately cloned into pBSKneo U6 and pGEM-T vectors. Ribozymes and caspase-7 mRNA were obtained by transcription in vitro, and ribozymes cleavage activity was identified by cleavage experiment in vitro.

RESULTSTwo ribozymes named Rz333 and Rz394 targeting 333 and 394 sites in caspase-7 mRNA were designed by computer software, and their DNA sequences were synthesized. The expression vector of caspase-7 and plasmids containing Rz333 and Rz394 were reconstructed successfully. Ribozymes and caspase-7 mRNA were expressed by in vitro transcription. In vitro cleavage experiments showed that Rz333 cleaved caspase-7 mRNA and produced 243nt and 744nt segments. The cleavage efficiency is 67.98%, while Rz394 cannot cleave caspase-7 mRNA.

CONCLUSIONSRz333 can site-specifically cleave caspase-7 mRNA.

Animals ; Base Sequence ; Caspase 7 ; Caspase Inhibitors ; Caspases ; genetics ; Cloning, Molecular ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; RNA, Catalytic ; biosynthesis ; genetics ; metabolism ; RNA, Messenger ; biosynthesis ; genetics

Objective To investigate the effects of physical exercises on cardiac function and plasma N-terminal pro-brain natriuretic peptide (NT-proBNP) levels in hypertensive patients combined with diastolic cardiac dysfunction.MethodsA total of 66 essential hypertension patients who had abnormal left ventricular relaxation and normal systolic function were assigned to the intervention group ( n =33 ; doing physical exercises once a day,5 days a week) or control group (n =33 ).All the patients received standard treatment.At 6 months,body weight,blood pressure,heart rate,NT-proBNP,and echocardiography were measured.ResultsAt 6 months,body weight [ (68 ± 7 ) kg vs (72 ± 8 ) kg ],systolic blood pressure [ (135.4 ±5.1) mm Hg (1 mm Hg =0.133 kPa) vs (141.9 ±5.2) mm Hg ],diastolic blood pressure [ (81.1 ±4.0) mm Hg vs (84.7 ±4.6) mm Hg],New York Heart Association class (1.4 ±0.3 vs 1.8 ±0.4),NT-proBNP level [ (526 ± 126 ) ng/L vs (741 ± 189 ) ng/L] were significantly decreased in the intervention group when compared with the control group ( all P ＜ 0.05 ) although left ventricular ejection fraction (LVEF) (62.9 ±6.7 vs 59.0 ±5.6) and E/A ratio ( 1.1 ±0.3 vs 0.9 ±0.3) were significantly increased ( both P ＜ 0.05).ConclusionPhysical exercises could play a role in reduced blood pressure and body weight and improved cardiac function in hypertensive patients with diastolic cardiac dysfunction.

Objective To evaluate the role of nutritional parameters in prognosis,especially in the early and late mortality of hospital-acquired acute kidney injury (AKI) patients.Methods This study was a prospective cohort study conducted in a hospital comprising 1500 beds in Shanghai, China. One hundred ninety-four patients with hospital-acquired AKI, as determined using the RIFLE staging criteria,were enrolled as subjects after obtaining informed consent.Patients with AKI caused by postrenal obstruction,glomerulonephritis,interstitial nephritis or vasculitis were excluded.Nutritional evaluation,including subjective global assessment (SGA),anthropometric and laboratory examination,was conducted. Other laboratory measurements and clinical data were recorded.The primary outcome was early mortality (≤ 7 days) and late mortality (8-28 days) after enrolling into the study. Results AKI patients at enrollment were characterized by a high prevalence of malnutrition as determined by SGA, anthropometric and laboratory examination.Univariate analysis showed that the SGA,the serum levels of prealbumin,cholesterol and total lymphatic cells, and the Maastricht index were significantly different among early mortality,late mortality and survival groups.The serum prealbumin and cholesterol levels in the early death group were significantly lower than those in the survival and late death groups (P＜0.05).Multivariate analysis revealed that SGA,albumin,prealbumin and cholesterol remained independently and significantly associated with early mortality after adjusting for age,sex,dialysis,ventilation,hemoglobin,platelets,bilirubin,and Glasgow coma score.The areas under the receiver operating characteristic curve to predict early mortality for albumin,prealbumin and cholesterol were 0.591,0.736 and 0.603,respectively,with that of prealbumin significantly higher than others (P＜0.05). Conclusion Low levels of serum prealbumin,albumin and cholesterol at enrollment are independtly associated with increased early mortality in hospital-acquired AKI patients.