Adult ; Albinism ; genetics ; Female ; Humans ; Male ; Middle Aged

[Objective]To discuss the results of the GIEBEL system in high tibial osteotomy to treat varus knee.[Method]Nineteen patients with medium varus knee were selected to be treated with osteotomy.Preoperatively,the mean varus angle was 12.29?,without deformity in other department of the knee joint.The main symptom was the pain in medial department.Accurate high tibial osteotomy with GIEBEL system fixation was performed in 31 knees of these patients.Weight-bearing extremity X-ray testing,varus angle,mechanical axle and anatomical axle were marked preoperatively and postoperatively.Joint space discrepancy between lateral and medial joint,the range of motion,Lysholm assessment and objective satisfactory survey were also performed to determine the outcome of the treatment.[Result]The mean follow-up of nineteen patients was 22 months.During this period,one case of temporary common fibular nerve paralysis occurred and healed at several days.All of the osteomy interface achieved bony healing.The corrected abnormality was 12.32?.The mechanism axle of the extremity maintained as the level just after operation during the follow-up and no degeneration of the joint space aggravated.The excellent and good rate of the treatment was 89.5%,and 84.2% of the patients expressed satisfactory for the operation.No difference of the range of motion was found before and after operation,but the Lysholm score,joint space discrepancy between lateral and medial joint and varus angle were improved after the osteotomy.[Conclusion]The GIEBEL system can fixate the bony interface of the high tibial osteotomy effectively,and possess the advantages of minimal invasion,stronger intensity and higher healing rate.

OBJECTIVE: To provide reference for reducing disadvantageous drug-drug interaction(DDI)and avoiding adverse drug event(ADE). METHODS: The patients aged more than 65 were selected from cardiovascular department in our hospital dur-ing Jun. 2015-Mar. 2016. The influential factors for potential DDI(PDDI)-induced ADE were analyzed. The relationship of related factors with PDDI-induced ADE was analyzed by multivariate Logistic regression analysis. RESULTS:A total of 328 patients were included,involving 257 PDDI patients,and totally 452 cases of PDDI (including 247 cases of mild PDDI,149 cases of general PDDI and 56 cases of severe PDDI). The age,the number of drugs used simultaneously,Ccr and liver function (Child-Pugh score)were related to the occurrence of PDDI-induced ADE(P<0.01). CONCLUSIONS:For PDDI-induced ADE,the risk evalua-tion can be conducted for a series of factors,including age,the number of drugs used simultaneously,Ccr and liver function. For high-risk patients,intervene should be conducted in advance to reduce the risk of ADE.

Objective To establish a model of full-thickness avascular meniscal defect to assess outcome of bone-marrow mesenchymal stem cells (BMSCs) modified with human insulin-like growth factor Ⅰ (hIGF-Ⅰ) gene and compounded with injectable calcium alginate gel in repair of meniscal defect.Methods Models of full-thickness defect were created in the anterior comer of meniscus in goats,an area lacking of blood supply.The trial categorized the models to four groups:gene-ehanced tissue engineering (GETE) group (hIGF-Ⅰ transfected BMSCs were mixed with calcium alginate gel),BMSCs group (BMSCs were mixed with calcium alginate gel),empty group (calcium alginate gel was used alone) and control group (the defect was excluded from repair).Macroscopy was done at 4,8,and 16 weeks after operation.Variation of repair tissue was observed by light and scanning electric microscopy and aggrecan in repair tissue was determined as well.Results Meniscal defect was on the mend at 4-16 weeks after operation in GETE group,with the defect area being thoroughly filled with the white,elastic and tight repair tissue similar to normal meniscal tissue.Macroscopic examination showed a better result in GETE group than that in other groups.Light microscopy showed that repair tissue which was mainly fibrochondrocytes was arranged in line with calcium alginate fibers and that space between the fibers was mostly crammed with the matrix secreted by those cells.At the same time,those cells were tightly arranged and the matrix secreted by those cells was equally distributed according to light microscopy.Electroscopy demonstrated neat and tight arrangement of fibers and tight extracellular matrix in fiber space in GETE group.Aggrecan concentration in GETE group was relatively higher than in other groups,but still had difference from the normal meniscus.Conclusion hIGF-Ⅰ gene-transfected BMSCs combined with injectable calcium alginate gel can improve the effect in repair of full-thickness meniscal defect.

BACKGROUND: Bone morphogenetic protein (BMP) and its derived scaffold or vector here been widely used in experiment and gradually applied in the clinic. However, the tracing method challenges the therapeutic effect, The discovery and application of enhanced green fluorescent protein (EGFP) can solve this problem.OBJECTIVE: To construct the bicistronic eukaryotic vector pIRES2-EGFP-hBMP-2 and to observe its expression in human embryo kidney (HEK) 293 cells.METHODS: hMBP-2 gene was extracted from human osteosarcoma by RT-PCR and inserted into PMD18-T vector. Following the DNA sequence verification, it was then sub-cloned into the eukaryotic vector pIRES2-EGFP. After restriction enzyme analysis the pIRES2-EGFP-BMP-2 was transfected into HEK 293 cells. Then its expression was observed using fluorescence microscopy and Western blot.RESULTS AND CONCLUSION: hBMP-2 gene was amplified and the eukaryotic vector plRES2-EGFP-hBMP-2 was constructed successfully. At 48 hours after tranfection of pIRES2-EGFP-hBMP-2 into HEK 239 cells, approximately 30% of cells emitted green fluorescence under a fluorescence microscope. Western blot results demonstrated that there was a specific BMP strap at Mr46×10s side, which indicated that the target gene could be expressed in constructed pIRES2-EGFP-hBMP-2 vector.

BACKGROUND: The existed repair method for cartilage defects has shortcomings of insufficient repairing tissue numbers, poor biomechanical properties, as well as donor site complication. Thus it is deficient to repair large-sized osteochondral defects using one method.OBJECTIVE: To investigate the therapeutic effect of tissue engineering modified Mosaicplasty on repairing large-sized osteochondral defects.DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed at the Center Laboratory of Qingdao University Medical College from January to September 2009.MATERIALS: The hircine bone mesenchymal stem cells (BMSCs) were in vitro cultured, and resuspended with algin solution to obtain BMSCs-calcium alginate gel.METHODS: Totally 12 goats were prepared for osteochondral defects models and were divided into 3 groups.BMSCs-Mosaicplasty group, BMSCs compound with injectable alginate calcium gel was then applied to fill the "dead space" after Mosaicplasty. In the Mosaicplasty group, the defects were repaired by Mosaicplasty. There was no treatment in the control group.From 4 to 16 weeks postoperatively, the animals were sacrificed and the in gross and under electromicroscopy.MAIN OUTCOME MEASURES: ①Gross observation: the joint was exposed to observe the repair effect at weeks 4, 8,16 after operation. ②Histological examination: specimens were harvested at 16 weeks after operation and observed by haematoxylin-eosin staining, toluidine blue staining under light microscopy. ③Transmission electron microscope was used at 16 weeks after operation.RESULTS: The transplanted subchondral bone and superficial cartilage was integrated hardly with each other or with recipient sites in tissue engineering modified Mosaicplasty groups at 16 weeks after operation. The quality and appearance of the transplanted and regenerated cartilage was similar to normal hyaline cartilage. Under microscopy, the regenerated cartilage was integrated with neighbor tightly in regular arrange. ECM distributed evenly and deeply stained by alcian blue. There was no obviously repaired in the control group.CONCLUSION: Tissue engineering can ameliorate the outcome of Mosaicplasty to repair the osteochondral defects.

Objective To investigate the effects of aggrecanase-2 knockdown in chondrocytes from rheumatoid arthritis patient by shRNA infection.Methods Cartilage harvested from rheumatoid arthritis patients who underwent total knee arthroplasty was digested by pancreatin and type Ⅱ collagen enzyme to obtain chondrocytes.Then chondrocytes were cultured and passaged to second or third generation.After aggrecanase-2 shRNA5,aggrecanase-2 shRNA6,aggrecanase-2 shRNA7,aggrecanase-2 shRNA8 infection,growth and morphological changes of the chondrocytes were examined.To select the best target sequence,mRNA expression of aggrecanase-2 and aggrecan was detected by RT-qPCR assay on day 2,5,10,represented with Ct (Cycle threshold) value.Expression of aggrecan protein was detected by immunocytochemistry,represented with IOD (integral optical density) value.Results aggrecanase-2 knockdown had no obvious effects on the morphology and growth of the chondrocytes.MOI (multiplicity of infection) was 100,50,25,and infection efficiency was 90％,60％,30％ with the corresponding viral load of 200 μl,100 μl,50 μl after transfection.mRNA expression of aggrecanase-2 was suppressed significantly,especially the group of shRNA5 which suppressed aggrecanase-2 expression from 0.876 3±0.115 6 to 0.069 9±0.015 1 (P ＜ 0.05).mRNA and protein expression of aggrecan were significantly upregulated after infection.mRNA expression of aggrecan increased from 0.992 1±0.201 3 to 3.049 2±0.278 2 (P ＜ 0.05) and protein expression of aggrecan increased from 496.160 5± 225.673 7 to 4 525.433 0±1 131.813 0 (P ＜ 0.05).Conclusion aggrecanase-2 suppression in chondrocytes by lentivirius infection is an effective method to protect the expression of aggrecan.

BACKGROUND: Cyclooxygenase 2, aggrecanase 1, and insulin-like growth factor 1 are involved in pathological injury of the articular cartilage. OBJECTIVE:To observe the expression of shRNA vectors carrying cyclooxygenase 2, aggrecanase 1 and overexpression vectors carrying insulin-like growth factor 1 in bone marrow mesenchymal stem cels. METHODS:Lentiviral vectors carrying the silencing gene cyclooxygenase 2, aggrecanase 1, the over-expressing gene insulin-like growth factor 1 and binding green fluorescent protein were constructed with recombinant lentiviral technology, and then the recombinant lentiviral vectors were used to transfect passage 3 human bone marrow mesenchymal stem cels culturedin vitro (experimental group). The human bone marrow mesenchymal stem cels transfected with no target gene lentivirals were used as negative control group. The human bone marrow mesenchymal stem cels transfected with no treatment served as blank group. RESULTS AND CONCLUSION:Cyclooxygenase 2 and aggrecanase 1 transfected in human bone marrow mesenchymal stem cels were significantly inhibited at gene and protein levels, while the expression of insulin-like growth factor 1 was increased significantly at gene and protein levels. We confirmed that cyclooxygenase 2 and aggrecanase 1 were successfuly silenced while insulin-like growth factor 1 overexpressed by using lentiviral vectors in human bone marrow mesenchymal stem cels, which brings a new hope for the systemic gene treatment of arthritis.

Objective To develop a method for detecting the genotype of Plasmodium vivax merozoite surface protein 1 (PvMSP-1) alleles. Methods According to the sequence characteristic of PvMSP-1, nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used to amplify the polymorphic region of ICB5-ICB6 which contains Q repeats and PvuII restriction site (Sal-1 type). The PCR product was digested by PvuII restriction endonuclease and the digested fragments were observed by 2% agarose gel electrophoresis. The allelic type was determined according to the banding pattern. Results Bands in size of 400 bp (Belem type ) and/or 470 bp (Sal-1 type ) appeared in all 98 P. vivax isolates, no band was found in negative control. After PvuII digestion, two Sal-1 type fragments (120 bp and 350 bp) were obtained from 45 samples of 470 bp. Single-band of 400 bp appeared in 3 of 40 samples with 400 bp as Belem type, two bands of 120 bp and 280 bp appeared from other 35 samples as recombination type III, and another 2 bands with 120 bp and 240 bp as Korean isolate. Conclusion The result showed that the nested PCR-RFLP may be applied in the detection and identification of the three PvMSP-1 allelic types in China.

Objective To evaluate the role of p38 mitogen-activated protein kinase (p38MAPK)-heat shock protein 27 (HSP27) signaling pathway in attenuation of lipopolysaccharide (LPS)-induced acute lung injury (ALl) by dexmedetomidine in mice.Methods Forty male Kunming mice,aged 2 months,weighing 20-25 g,were equally and randomly divided into 4 groups using a random number table:control group (group C),LPS group,low-dose dexmedetomidine + LPS group (group D1),and high-dose dexmedetomidine + LPS group (group D2).Dexmedetomidine 25 and 50 μg/kg were injected intraperitoneally in D1and D2 groups,respectively,and 1 h later LPS 5 mg/kg was injected intraperitoneally.At 6 h after LPS injection,the left lung was lavaged,and broncho-alveolar lavage fluid (BALF) was collected for determination of concentrations of protein,tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β).The right lung was removed for examination of the pathological changes (under the light microscope) and for detection of expression of phosphorylation of p38MAPK (p-p38MAPK),p38MAPK,phosphorylation of MAPK-activated protein kinase 2 (p-MAPKAPK-2),MAPKAPK-2,phosphorylation of HSP27 (p-HSP27) and HSP27 in lung tissues.The wet to dry lung weight (W/D) ratio was calculated.The ratios of p-p38MAPK/p38MAPK,p-MAPKAPK-2/MAPKAPK-2 and p-HSP27/HSP27 were calculated.Results Compared with group C,the W/D ratio,concentrations of protein,TNF-α and IL-1β in the BALF,and ratios of p-p38MAPK/p38MAPK,p-MAPKAPK-2/MAPKAPK-2 and p-HSP27/HSP27 were significantly increased in group LPS.Compared with group LPS,the W/D ratio,concentrations of protein,TNF-α and IL-1β in the BALF,and ratios of p-p38MAPK/p38MAPK,p-MAPKAPK-2/MAPKAPK-2 and p-HSP27/ HSP27 were significantly decreased in D1 and D2groups.The pathological changes of the lung were significantly reduced in D1 and D2 groups as compared to LPS group.Conclusion Dexmedetomidine attenuates LPS-induced ALI in mice possibly through inhibiting p38MAPK-HSP27 signaling pathway.