Knee Osteoarthritis is one of the most common degenerative diseases.Although the morbidity of knee osteoarthritis increases dramatically,there is no effective medication that can repair the damaged articular cartilage and restore the joint function.Recently,with the technical development of cell transplantation and regulation,more attentions have been paid in this area.The techniques included osteochondral autograft transplantation,mosaicplasty,chondrocytes based matrix-induced autologous chondrocyte implantation,and mesenchyma stem cells based cell therapy.The study of cell differentiation is also more in-depth.It has been confirmed that all of these methods can alleviate the symptoms and improve the function of knee osteoarthritis to some extent.Stem cell therapy seems have greater advantage than other intervention.Massive chondrocytes can be gained through induction and culture of mesenchyma stem cells in vitro,and regenerated cartilage could be found on the surface after delivering stem cells into knee joint through a certain way.These technologies are hopeful to prevent from knee osteoarthritis or even reverse the progress of it at early stage.However,there are a variety of problems about in vitro cell transplantation,such as the unknowing regulation of mesenchyma stem cells,the difficulty of keeping the characteristic stability of new born chondrocytes,the integrity of new cartilage tissue and the way chondrocytes delivered.Cell therapy has been applied both in basic research and in clinic trials.The present review is focused in the current progress of cell therapy in knee osteoarthritis and discuss the challenges and troubles.Furthermore,we also summarize a series of ongoing clinical trials,and try to find out the exact clinical effects of cartilage repair by using cell therapy.

BACKGROUND: Multipotent differentiation ability enables mesenchymal stem cells from autologous bone marrow to differentiate into osteoblasts and chondroblasts, thereby promoting the formation of bones and cartilage. However, the osteogenic ability differs from each other, and whose osteogenic ability is the best still needs to be studied further. OBJECTIVE: To compare the osteogenic ability of fascia- and muscle-derived stem cells in rats. METHODS: Fascia- and muscle-derived cells were isolated from 20 rats, followed by flow cytometry sorting, and were then cultured. FDC-LacZ cells were transfected with retro-BMP4 virus twice. Afterwards, the transfection efficiency of fascia-derived cells was detected through LacZ and alkaline phosphatase staining. RESULTS AND CONCLUSION: Compared with fascia-derived cells, muscle-derived cells showed stronger chondrogenic ability and produced more calcium deposition. These findings indicate that the osteogenic ability of muscle-derived cells is superior to that of fascia-derived cells in rats.

Objective To produce bioactive human bone morphogenetic protein-4 by Escherichia coli genetic engineering and investigate the effect of the product, recombinant BMP-4, on the bone marrow stem cells. Methods cDNA of human morphogenetic protein-4 mature peptide was obtained by RT PCR from tissue of human placenta. The gene was constructed in the pET-22b(+) expression vector and expressed in Escherichia coli BL-21 after transformation and induction by IPTG. The harvested protein was proved to be bioactive by inducing ectopic bone information in mouse thigh. The protein was applied to induce the cultured bone marrow stem cell. Shape change of the cell, ability of ALP (alkaline phosphatase) and concentration of OC (osteocalcin) were investigated. Results SDS-PAGE revealed a new protein band that located in position of 14?103 after 4 hours induction, the new protein made 15% of total bacteria protein, the rhBMP-4 could induce the cultured bone marrow stem cell of New Zealand rabbit to differentiate into osteoblasts and form calcified node. The ability of ALP and concentration of OC of tested group increased significantly than that of the control group. Conclusion The bioactive rhBMP-4 can be produced by Escherichia coli genic engineering, this protein can induce bone marrow stem cells to differentiate into osteoblast cells.

BACKGROUND:Mesenchymal stem cells from human skeletal muscle exhibit multi-directional differentiation potential under the influence of osteogenic proteins such as bone morphogenetic protein 4 (BMP4). But the differentiation of a specific cell subpopulation is not yet clear.OBJECTIVE:To characterize the multi-directional differentiation potential of mesenchymal stem cells from human skeletal muscle based on the expression of different surface markers.METHODS:Four different subpopulations were isolated from the human skeletal muscle by fluorescence-activated cell sorting based on their expression of the myogenic-specific marker CD56 and the endothelial-specific markers CD34 and CD144, including CD56+, CD56+CD34+CD144+, CD34+CD144+, and unsorted groups. Osteogenic differentiation of the four groups of the cells was displayed by Von Kossa staining after the treatment with BMP4 alone or BMP4 plus transforming growth factor β3. Chondrogenic differentiation of these cells was displayed by Alcian blue staining. Bone metabolism was assessed by alkaline phosphatase staining.RESULTS AND CONCLUSION:No significant difference in the bone metabolism was found among four groups after the treatment with BMP4 (P > 0.05). Osteogenic and chondrogenic potentials of the four cell subpopulations were significantly different. Under the same osteogenic induction, the CD56+ cells exhibited strongest potential for osteogenic differentiation; and under the same chondrogenic induction, the CD56+CD34+CD144+ cells exhibited better potential for chondrogenic differentiation than the CD56+ cells. These findings indicate that the osteogenic and chondrogenic potentials are intimately associated with the type of mesenchymal stem cells from human skeletal muscle:the CD56+ cells are closely related to the osteogenic potential, while the CD56+CD34+CD144+ cells have stronger chondrogenic potential.

Objective To explore whether anoxia can induce expression changes in connective tissue growth factor(CTGF)in renal tubular epithelial cells(TECs)and epithelial-mesenchymal transition of TECs.Methods Rat renal TECs(NRK-52E)anoxia models were established.NRK-52E cells were exposed to anoxia for 4 h.The real-time RT-PCR,Western blotting,immunohistochemical staining were used to detect the expression of CTGF at 6,12,24,48,and 72 h in NRK-52E cells.Morphological changes and cytoskeleton remodeling in NRK-52E cells under anoxia were examined by a laser confocal microscope and BODIPYFL staining respectively.Results Under anoxia,NRK-52E cells became round,enlarged and cytoskeleton was remodeled.The expression levels of CTGF mRNA and protein were up-regulated at 6 h,reached their peak at 48 h:the expression of CTGF mRNA protein was 29.33±0.21 and 1.30±0.02 respectively.Under anoxia,NRK-52E cells underwent an epithelial-mesenchymal transition process,including cytoskeleton remodeling,and morphological changes.Conclusion Anoxia can change the expression of CTGF and other fibrosis-associated genes in NRK-52E cells,and CTGF played an important role in fibrosis process and epithelial-mesenchymal transition development in NRK-52E cells.

Objective To assess the value of functional magnetic resonance urography (fMRU)for the unilateral renal function in children with hydronephrosis.Methods Fourteen children with congenital hydronephrosis (unilateral hydronephrosis in 1 2 cases,bilateral hydronephrosis in 2 cases)examined by fMRU in Beijing Children′s Hospital,Capital Medical University,were enrolled.In 7 patients of them,diuretic renal scintigraphy (DRS)was per-formed within 1 0 days before fMRU examination.The following parameters in fMRU,as renal parenchymal volume,volu-metric differential renal function (vDRF),Patlak,Patlak differential renal function (pDRF),index of glomerular filtra-tion rate (GFR)and differential renal function based on index of GFR (gDRF),were calculated and analyzed.Statisti-cal analysis was performed by using SPSS 1 3.0.Results In 7 cases whose fMRU and DRS were examined,the indexes of GFR obtained from fMRU and GFR from DRS were well correlated (r =0.892,P <0.001 )in 1 4 kidneys.The gDRF determined by 2 methods on the left kidneys[the average was(46.80 ±1 9.20)% and(45.1 8 ±20.29)%,respective-ly]had no significant difference (t =0.051 6,P =0.624),which was also highly correlated (r =0.91 2,P =0.004). In 1 2 cases with unilateral hydronephrosis,vDRF,pDRF,index of GFR and gDRF in hydronephrotic side[(43.54 ± 9.61 )%,(42.80 ±1 0.83)%,(38.56 ±29.23)mL/min,(38.37 ±1 3.61 )%]were all less than those in the con-tralateral side[(56.46 ±9.61 )%,(57.1 9 ±1 0.83)%,(57.02 ±26.22)mL/min,(61 .63 ±1 3.61 )%](t =2.326, 2.300,2.422,2.960;P =0.040,0.042,0.034,0.01 3).However,there was no statistical difference in both renal pa-renchymal volume and Patlak between the hydronephrotic and the contralateral side kidneys(t =1 .765,1 .450;P =0.1 05,0.1 75).Conclusions fMRU is a very valuable examination method in evaluating single kidney function in children with congenital hydronephrosis,and able to demonstrate that gDRF,indexes of GFR,vDRF and pDRF decrease in the hydronephrotic kidney.

Post competency is a kind of necessary ability for professional working.In order to train the students' excellent professional skills,we have conducted exploration and reform in post competency oriented experiment teaching,through reforming experimental courses,changing teaching methods and means of examination,constructing the laboratory etc.,so that we have built a new professional training model in pathological diagnosis and technology specialty experimental teaching.The experimental teaching reform results have improved the experimental teaching quality and the talents capability.

Objective To investigate the diagnostic value of MRI in reversible splenial lesion syndrome (RESLES) in children.Methods The clinical and MRI imaging data of 14 cases of RESLES in children were retrospectively reviewed.There were 4 males and 10 females,aging 11 to 35 months.Average age was (20±3) months.MRI studies were conducted in all the cases.Two experienced doctors analyzed independently the images and reached consensus.Results Fourteen cases showed the single abnormal signal in the splenial of corpus callosum on the initial cerebral MRI.Lesions in 10 of 14 cases appeared as isointensity on T1WI,hypo-intensity in 4 cases.Lesions in 14 cases were appeared as hyper-intensity on T2WI,hyper-/slightly hyper-intensity on FLAIR T2WI.The lesions were round,oval or irregular,with fuzzy boundary.DWI showed round or oval high signal with clear boundary.No edema around the lesion and no occupying effect were detected.After treatment,8 of 14 cases were reviewed after 7-14 d,and the abnormal signal in the corpus callosum disappeared in 5 cases,while the range was significantly reduced in 3 cases.MRI were reexamined after 1-2 months in 5 cases,and the lesions disappeared in the corpus callosum.One case didn't have MRI examination again after treatment and the clinical symptoms disappeared.Conclusions The clinical manifestations of RESLES in children are lack of specificity.And the cerebral MRI imaging is characterized by the reversible solitary lesion of the splenium,which can provide a reliable basis for clinical diagnosis and prognosis.

Objective To evaluated the clinical value of laparoscopic techniques in radical cystectomy surgery for the treatment of bladder cancer. Methods Clinical data of 49 patients underwent radical cystectomy with Bricker ileal conduit diversion were retrospectively analyzed from October 2009 to August 2014, which laparoscopic radical cystectomy with Bricker ileal conduit 20 cases (Group A), open radical cystectomy with Bricker ileal conduit 29 cas-es (Group B). The blood loss during operation, operating time, gastrointestinal function recovery after operation, hos-pital stay after operation and complications were observed between the two groups. Results The blood loss during operation was significantly lower in Group A (416.66 ± 232.73) ml than in Group B (964.16 ± 445.73) ml ( <0.05), and hospital stay after operation was significantly lower in Group A (14.93 ± 2.72) days than in Group B (19.50 ± 3.16) days ( < 0.05), complication after operation was significantly lower in Group A than in Group B ( < 0.05). The operating time and gastrointestinal function recovery has no significantly difference between the two groups. Conclusions Laparoscopic radical cystectomy have advantages of minimal invasion, less blood loss, rapid recovery and less postoperative complications. It is a safe and effective surgical method. Long term effect need evaluated by follow up.

BACKGROUND: Osteoarthritis is a common cause of pain and disability in the elderly. The underlying cause is the combination of inappropriate mechanical stress, inflammatory mediators and biochemical factors. Curcumin has been shown to have anti-oxidant, anti-inflammatory, anti-bacterial, anti-tumor, neuroprotective, and cardioprotective effects, radiation protection and therapeutic effects on osteoarthritis. Similarly, magnesium sulfate can relieve joint pain and inhibit joint destruction. OBJECTIVE: To investigate the mechanism by which the combined use of curcumin and magnesium sulfate exerts synergistic effect to promote inflammatory chondrocyte reverse differentiation.METHODS: Primary cultured inflammatory chondrocytes were subjected to monolayer culture in vivo. Cell proliferation assay (MTS) was used to detect the proliferation of inflammatory chondrocytes under in vitro monolayer culture conditions. The experimental cells were divided into four groups and underwent 3D induced reverse differentiation culture for 18 days: single culture of chondrocytes (chondrocyte group) , inflammatory chondrocyte cultured with curcumin (curcumin group) , inflammatory chondrocytes cultured with magnesium sulfate (magnesium sulfate group) , and inflammatory chondrocytes cultured with curcumin and magnesium sulfate (combination group). RESULTS AND CONCLUSION: MTS proliferation experiments showed that inflammatory chondrocytes at passage 3 had a higher rate of early proliferation and a lower degree of differentiation. Quantitative PCR results showed that the mRNA levels of type II collagen, proteoglycan and SOX9 were significantly higher in the combination group than in the curcumin group or magnesium sulfate group (P < 0.01). The size of the gross specimens and the positive area of chondrocyte reverse differentiation for alcian blue staining and safranin O staining in the combination group were significantly larger than those in the other three groups (P < 0.01). TUNEL staining results indicated that the positive area of apoptosis-specific staining in the combination group and magnesium sulfate group was significantly smaller than that in the other two groups (P < 0.01). Therefore, the combined use of curcumin and magnesium sulfate has the synergistic effect to promote the reverse differentiation of inflammatory chondrocytes.