Objective:To investigate effects of bone-resorptive lesion on stress distribution of femoral head and on progression in patients with osteonecrosis of the femoral head (ONFH).Methods:From April 2014 to September 2018, a total of 155 femoral heads from 94 patients diagnosed with ARCO stage II and III ONFH were retrospectively reviewed, including 77 males and 17 females with aged 39.90±10.45 years old (ranged from 18-64 years). The hips were divided into two groups according to whether there were bone-resorptive lesions. Further, we compared whether there was statistical difference between the two groups in staging. Then, a case of ARCO II hip joint without bone-resorptive lesion was selected from the included patients. Six femoral head with different diameters of spherical bone-resorptive lesion of 5 mm, 7 mm, 10 mm, 14 mm, 18 mm, and 23 mm were simulated. The influence of bone-resorptive lesion on the stress distribution of necrotic area and a spherical shell extending 1 mm radially around the bone-resorptive lesion was investigated by finite element method in slow walking conditions.Results:Of the 155 ONFH hips, 67 hips are complicated by bone-resorptive lesions, of which 17 were ARCO II, 50 were ARCO III. A total of 88 hips did not contain bone-resorptive lesions, of which 58 were ARCO II, ARCO III 30 cases. The proportion of ARCO stage II in the group with bone-resorptive lesions was significantly higher than that in the group without bone-resorptive lesions (χ 2=25.03, P=0.000). The finite element stress distribution cloud diagram showed that there was a stress concentration area around the bone-resorptive lesions. The maximum von Mises stress around bone-resorptive lesions in the models that contained a synthetic bone-resorptive lesions were significantly higher than those reported in the matched, non-synthetic bone-resorptive lesions finite element models ( t=3.139, P=0.026). The values for maximum von Mises stress around bone-resorptive lesions were 6.94±1.78 MPa and 5.01±0.35 MPa for the group with synthetic bone-resorptive lesions and the group non-synthetic bone-resorptive lesions, respectively. There was a positive correlation between the diameter of bone-resorptive lesions and the maximum and mean von Mises stress of necrotic areas as well as the maximum von Mises stress around bone-resorptive lesions. Conclusion:Bone-resorptive lesions can increase the maximum stress and average stress in the necrotic area. The larger the bone-resorptive lesion, the more the stress increases. There is a stress concentration area around the bone-resorptive lesions, which may accelerate the collapse of the femoral head.

Objevtive To investigate the efficacy of Xuebijing injection combined with Ulinastatin for acute pancreatitis.Methods Databases were searched,like Pubmed,Embase,Cochrane Central Register of Controlled Trials (CENTRAL),China Biology Medicine disc (CBM),China National Knowledge Infrastructure (CNKI),Cochrane library,and Wangfang for randomized controlled trial (RCT) about the treatment of Xuebijing injection combined with Ulinastatin for acute pancreatitis.After evaluating the quality of literatures objectively,data were analyzed by RevMan 5.0 software.we evaluated abdominal pain relief time,recovery time of blood amylase,recovery time of white blood cell (WBC),concentration of interleukin (IL)-6,IL-8,tumor necrosis factor α (TNF-o) and total effective rate.Results Eleven studies and 893 patients were accepted into this article.Meta-analysis showed that abdominal pain relief time [weighted mean difference (WMD) =-1.71,95 ％ CI:-2.21,-1.21,P ＜ 0.01],recovery time of blood amylase (WMD =-1.82,95 ％ CI:-2.39,-1.25,P ＜ 0.01),recovery time of WBC (WMD =-2.75,95 ％ CI:-3.19,-2.31,P ＜ 0.01),and hospital stay time (WMD =-5.99,95 ％ CI:-7.73,-4.26,P ＜ 0.01)in experimental group was better than control group.Compared to control group,on the seventh day after treatment,inflammatory cytokines,including IL-6 [standardized mean difference (SMD) =-1.09,95％ CI:-2.66,0.48,P=0.17],IL-8 (SMD=-1.02,95％ CI:-1.66,-0.38,P＜0.01),andTNF-α (SMD =-1.10,95％ CI:-1.68,-0.53,P ＜ 0.01) were lower.In experimemal group,total effective rate was better than the control group (RR =1.16,95％ CI:1.07,1.25,P=0.0002).Conclusions Xuebijing injection combined with Ulinastatin for acute pancreatitis was more effective than traditional basal treatment or using Ulinastatin alone.However,the literature quality were mediocre,we need more large,random,double blind,and polycentric clinical study to prove further.

BACKGROUND:Accumulative evidence supports that co-culture technology can be applied to construct the tissue-engineered cartilage with excellent biological characters. OBJECTIVE:To elaborate the co-culture concept and conclude and analyze seed cell sources, cel mixed ratio, spatial y-defined co-culture models and biomaterials in co-culture systems to conclude and analyze the biological characters of tissue-engineered cartilage, and to prospect progression of co-culture systems in cartilage tissue engineering. METHODS:The first author retrieved the databases of PubMed, Web of Science, and CNKI for relative papers published from January 1976 to May 2016 using the keywords ofco-culture, co-culture systems;articular cartilage, chondrocytes, mesenchymal stem cells;tissue engineering, articular cartilage tissue engineeringin English and Chinese, respectively. Finally 60 literatures were included in result analysis, including 1 Chinese and 59 English articles. RESULTS AND CONCLUSION:Co-culture technology emphasizes the role of microenvironment in terms of various physical, chemical and biological factors in the cell processing. In cartilage tissue engineering, co-culture systems contribute to maintain the viability and natural cell phenotype of chondrocytes and induce cartilage differentiation of mesenchymal stem cells. In addition, co-culture technology provides a novel way for cartilage tissue engineering to overcome the shortage of chondrocytes and repair injury to the cartilage-subchondral bone. However, the mechanisms of cell-cell interaction in co-culture systems still need to be explored in depth, so as to optimize the co-culturing conditions and construct perfect tissue-engineered cartilage.

Objective To determine the effect of NEL-like type 1 gene (NELL-1) transfection in vivo in the repair of traumatic femoral head necrosis.Methods Twenty-four SD rats were randomly divided into three groups (8 rats per group) according to the lottery method,ie,sham group (served as normal control),NELL-1 treatment group (injected NELL-1 gene by recombinant adenovirus vectors around the hip one week after osteonecrosis model induced surgically) and placebo group (given an equal volume of saline solution at the same time after the induction of osteonecrosis).Femurs were taken from the animals 5 weeks after surgery.Gross observation was performed for morphology changes,X-ray assessment for femoral head height and length ratio (H/L),Micro-CT measure for bone parameters of femoral head including total volume (TV),bone volume (BV),total mineralized content (TMC),trabecular thickness (Tb.Th) and trabecular space (Tb.SP),and histological study for osteocytes,osteoblasts and osteoclasts.Results Preserved femoral head shape was noted in NELL-1 treatment group compared to the obvious flattening of the femoral head in placebo group.No heterotopic osteogenesis was observed in any group.Femoral head H/L ratio for 0.753 2 ± 0.040 2 in NELL-1 treatment group was higher than 0.598 4 ± 0.037 0 in placebo group (P ＜ 0.05),but lower than 0.920 2 ± 0.037 0 in sham group (P＜0.05).TV,BV,TMC and BMD between NELL-1 treatment and sham groups did not differ significantly (P ＞ 0.05),but all were increased compared to placebo group (P ＜ 0.05).There was no significant differences in Tb.Th and Tb.SP among three groups (P ＞ 0.05).Most osteocytes were alive in NELL-1 treatment group.More active osteoblasts and osteoclasts were noted in NELL-1 treatment group than those in placebo group.Conclusion NELL-1 gene transfection can preserve femoral head shape and bone content,promote osteoblast activity and neovascularization and hence is an effective treatment for rat traumatic osteonecrosis.However,the activity of osteoclasts is stimulated simultaneously.

OBJECTIVE:To establish a method for the residues determination of Pb,Cd,Cu,As and Hg in Codonopsis pilo-sula,and evaluate the quality evaluation of C. pilosula of Pingshun County in Shanxi province. METHODS:Microwave diges-tion-inductively coupled plasma mass spectrometry was adopted with KED scanning model,RF power was 1 550 W,sampling depth was 5.0 mm,plasma gas(argon)flow rate was 16.0 L/min,helium partial pressure was 0.1 mbar,argon gas was 0.6 mbar, the vacuum degree of 5×10-8 mbar,branch turbopump speed was 1 000 hz,sampling cone aperture was 1.0 mm,skimmer aperture was 0.5 mm,the spray chamber temperature was 2.7 ℃,the data collection was repeated 3 times. RESULTS:The linear range was 0-20 ng/ml for Pb(r=0.999 3),0-10 ng/ml for Cd(r=0.998 5),0-250 ng/ml for Cu(r=0.998 8),0-20 ng/ml for As(r=0.999 0) and 0-1.0 ng/ml for Hg(r=0.997 9);RSDs of precision,stability and reproducibility tests were lower than 3.0%;recoveries were 95.80%-100.20%(RSD=1.85%,n=6),94.50%-98.00%(RSD=1.26%,n=6),98.52%-102.43%(RSD=1.60%,n=6), 94.90%-98.70%(RSD=2.29%,n=6)and 96.00%-101.00%(RSD=1.84%,n=6);the limits of detection were 0.021 0,0.003 4, 0.043 7,0.115 6 and 0.005 6 ng/kg,respectively. Pb,Cd,Cu,and As were detetcted,and Hg was not detected,the range of total contents was 7.185 2~12.558 0 mg/kg. CONCLUSIONS:The method is simple with good precision,stability and reproducibility, and can be used for the residues determination of Pb,Cd,Cu,As and Hg in C. pilosula;heavy metal residues in C. pilosula in Shanxi Pingshun county does not exceed limit values of national and industry standards.

BACKGROUND:Cartilage extracelular matrix with a large number of signaling molecule proteins and factors is likely to be an ideal material for tissue engineering cartilage.
OBJECTIVE: To investigate the possibility of calcium alginate and cartilage extracelular matrix combined with microencapsulated stem cels derived from human umbilical cord Wharton’s jely to construct ectopic tissue-engineered cartilage in nude mice.
METHODS: Microfilament suspension of the cartilage extracelular matrix was prepared. Human stem cels derived from Wharton’s jely of the umbilical cord were inoculated in to calcium alginate and cartilage extracelular matrix gel microspheres as experimental group. Stem cels derived from human umbilical cord Wharton’s jely were incubated in simple alginate gel microspheres as control group. After in vitro culture, the microspheres wereimplanted into the dorsal subcutaneous tissue of nude mice. Samples were taken after 4 weeks, respectively, for gross and histological observation.
RESULTS AND CONCLUSION:The stem cels exhibited paralel-chondrocyte morphology in microspheres, which grew and proliferated quite wel during in vitro culture. A new paralel-cartilaginous tissue was found in the subcutaneous tissue 4 weeks after surgery in the experimental group, and the tissue was positive for hematoxylin-eosin, safranine O, toluidine blue and colagen II. A large number of paralel-chondrocytes and cartilage lacuna-like structures were observed under a microscope with no obvious inflammatory reaction around the microspheres. The control group showed the partial degradation of microspheres, surrounded by only a smal number of inflammatory cels and lymphocytes. Calcium alginate and cartilage extracelular matrix microspheres have a rather good histocompatibility which can be used to construct paralel-cartilaginous tissues by implanting stem cel-microspheric compound into the subcutaneous tissue of nude mice.

Objective To explore the feasibility of fabricating a novel cartilage acellular matrix/chitosan hybrid scaffold for cartilage tissue engineering. Methods Human cartilage microfilaments about 100 nm-5 μm were prepared after pulverization and made into 1% suspension after decellularization. The suspension was mixed with 2% chitosan acetic acid solution, and then hybrid scaffolds were fabricated using a simple freeze-drying method. The scaffolds were cross-linked and were investigated by histological staining,SEM observation, porosity measurement, water absorption rate, biomechanical properties, and biocompatibility analysis. MTT test was also done to assess the cytotoxicity of scaffold leaching liquor. Canine chondrocytes were isolated and seeded into the scaffold. Cell proliferation and differentiation were analyzed using inverted microscope and SEM. Results The histological staining showed no chondrocyte fragments remained in the scaffolds, and anti-col Ⅱ immunohistochemistry staining were positive. SEM observation show the scaffold has good pore interconnectivity with pore diameter (136.2±34.9) μm, 81.4%±3.5% porosity and 1525.7%±129.3% water absorption rate. The longitudinal elastic modulus of the scaffold was (1.940±0.335) MPa. MTT test showed that the scaffold leaching liquor did not exert any cytotoxic effect on BMSCs. Inverted microscope and SEM micrographs indicatod that cells covered the scaffolds uniformly, and majority of the cells showed the round or elliptic morphology with much matrix secretion. Conclusion Novel cartilage acellular matrix/chitosan hybrid scaffold had similar extracellular matrix as cartilage, good pore diameter and porosity,appropriate biomechanical character, non-toxicity and good biocompatibility, which make it a suitable candidate as an alternative cell-carrier for cartilage tissue engineering.

Objective To probe the immunological traits of mesenchymal stem cells derived from umbilical cord Wharton's jelly (WJMSCs). Methods The diced Wharton's jelly which was from healthy fullterm birth human umbilical cord was cultured. The mesenchymal stem cells were identified with mesenchymal stem cells markers expression by flow cytometry and multiple differentiation ability. The expression of MHC- Ⅰ / Ⅱ, costimulatory molecules (CD40, CD80 and CD86) was detected with flow cytomctry, immunocytochemistry, and RT-PCR. The expression of immune inhibitors like HLA-G, IDO, and PGE2 was detected by immunocytochemistry and RT-PCR. The expression of immune-related molecules as IL-10, TGF-β, FGF and VEGF was detected with antibody microarray and western blot. Further more, to clarify the in vivo immune reaction of hWJMSCs, we fabricated the hWJMSC-scaffold constructs and implanted them into the rabbit backs. The lymphocyte infiltration and implanted cell survival observed with immunofluorescence. Results After culturinge of diced Wharton's jelly tissue, we obtained spindle-shaped cells. With differentiation medium, the cells can differentiate into osteoblasts, chongdrocytes, adipose cells and schwann cells. Expression of MHC, costimulatory molecules, and a series of immune suppressive-related molecules was found. Immune inhibitors as HLA-G, 1DO, PGE2, and immune suppressive related molecules as HGF, VEGF, TGFand IL-10 were positively expressed. But the cells did not express MHC-Ⅱ. No immune rejection was observed in vivo after implantation of hWJMSC-scaffold constructs. Conclusion It can be concluded that hWJMSCs have very low immunogenicity, which means the cells have potential to induce immune tolerance.The hWJMSCs do not provoke immune rejection in vivo.

Objective To investigate the effects of the novel scaffold on repairing large,high-loadbearing osteochondral defects of femoral head in a canine model.Methods The biphasic scaffolds were fabricated using cartilage extracellular matrix (ECM)-derived scaffold (cartilage layer) and acellular bone matrix (bone layer) by phase separation technique.Articular high-load-bearing osteochondral defects with a diameter of 11-mm and the depth of 10-mm were created in femoral heads.The defects were treated with constructs of a biphasic scaffold seeded with chondrogenically induced bone marrow-derived mesenehymal stem cells (BMSCs).The outcomes were evaluated for gross morphology,histological,biomechanical and micro-CT analysis at the third and sixth month after implantation.Results The gross and X-ray results showed femoral head slightly collapsed at the third month and severely collapse at the sixth month.Histological analysis showed cartilage defects were repaired with fibrous tissue or fibrocartilage with severe osteoarthritis and the varied degrees of the collapse of femoral heads were presented.Micro-CT showed that the values of bone volume fraction in defect area were always lower than those of the normal area in the femoral heads.Biomechanical analysis showed rigidity of the subchondral bone in defect area was significantly lower than that in normal area in the femoral heads at the sixth month.Conclusion The ECM-derived,integrated biphasic scaffold seeded with chondrogenically induced BMSCs could not successfully repair the large high-load-bearing osteochondral defects of the femoral head.

Objective To fabricate cartilage extracellular matrix (ECM) oriented scaffolds and investigate the attachment, proliferation, distribution and orientation of bone marrow mesenchymal stem cells (BMSCs) cultured within the scaffolds in vitro. Methods Cartilage slices were shattered in sterile phosphate-buffered saline (PBS) and the suspension were differentially centrifugated untill the micro- fiber of the cartilage extracellular matrix was disassociated from the residue cartilage fragments. At last the supernatant were centrifugated, the precipitation were collected and were made into 2%-3% suspension. Using unidirectional solidification as a freezing process and freeze-dried method, the cartilage extracellular matrix derived oriented scaffolds was fabricated. The scaffolds were then cross-linked by exposure to ultraviolet radiation and immersion in a carbodiimide solution. By light microscope and scan electron microscope (SEM) observation, histological staining, and biomechanical test, the traits of scaffolds were studied. After being labelled with PKH26 fluorescent dye, rabbit BMSCs were seeded onto the scaffolds. The attachment, proliferation and differentiation of the cells were analyzed using inverted fluorescent microscope. Results The histological staining showed that toluidine blue, safranin O, alcian blue and anti-collagen Ⅱ immunohistochemistry staining of the scaffolds were positive. A perpendicular pore-channel structures which has a diameter of 100 μm were verified by light microscope and SEM analysis. The cell-free scaffolds showed the compression moduli were (2.02±0.02) MPa in the mechanical testing. Inverted fluorescent microscope showed that most of the cells attached to the scaffold. Cells were found to be widely distributed within the scaffold, which acted as a columnar arrangement. The formation of a surface cells layer was found on the surface of the scaffolds which resembled natural cartilage. Coclusion The cartilage extracellular matrix derived oriented scaffolds have promising biological, structural, and mechanical properties.