Objective:To analysis the correlation of Wnt/β-catenin signaling pathway,OPN and MMP-13 in rabbit model with different degree of osteoarthritis (OA).Methods:Forty New Zealand white rabbits were randomly divided into four groups of 10 each,then we constructed the rabbit model with different degree of osteoarthritis (OA) by different concentrations of papain.The mRNA and protein levels of β-catenin,OPN,MMP-13,type Ⅱ collagen and proteoglycan were detected by Real-Time PCR and ELISA,respectively.Results:We have successfully established OA rabbit model by different concentrations of papain,and these OA rabbits model could be divided into mild,moderate and severe three level by Mankin scoring system,chondrocytes.Compared with the normal control group,the protein levels of β-catenin,OPN,and MMP-13 in the osteoarthritis were significantly increased,while the protein levels of type Ⅱ collagen and proteoglycan were significantly decreased (P＜0.05).With the increasing severity of OA,the results were consistent (P＜0.05).Conclusions:These results indicated that Wnt/β-catenin signaling pathway might regulate the expression of OPN,thereby affecting the expression of MMP-13,and ultimately have an impact on the occurrence ofosteoarthritis.

BACKGROUND:Seed cells from different sources have different ability in cell adhesion,proliferation,and differentiation,which can led to bioactive diversity in constructed tissue engineered products. OBJECTIVE:To explore the differentiation ability of different original fetal osteoblasts during constructing tissue engineered periosteum at molecular level. DESIGN,TIME AND SETTING:The contrast observation was performed at the Central Laboratory of Shaanxi Provincial Peoples’ Hospital between July 2007 and July 2008. MATERIALS:The human amnion cells(consent was obtained from the puerperant) were prepared human acellular amniotic membrane(HAAM) . METHODS:Periosteum-origin osteoblasts(POB) and cranium-origin osteoblasts(COB) were seed on HAAM,cultured for 2,4,6,8,and 10 days,and then their total RNA was extracted,which were reversely transcripted to cDNA. The real-time PCR analysis was used to reveal core binding factor ?l(Cbfa1) ,Osterix,and the cycle threshold was also measured. MAIN OUTCOME MEASURES:The expression of Cbfa1,Osterix,as well as osteocalcin. RESULTS:On tissue engineered periosteum,the expression of Cbfa1 in POB was lower than it that in COB(P

BACKGROUND:There are many researches on the finite element analysis of total hip arthroplasty, but the biomechanical distribution after hip surface replacement is little reported.OBJECTIVE: To analyze the biomechanical characteristics after hip surface replacement based on three- dimensional finite element analysis.METHODS: The three-dimensional finite element model after metal-to-metal hip surface replacement was established.The load of 350 N paralleling to the gravity line was loaded on the acetabulum. The stress distribution of the prosthesis and femur and the stress of the different regions of the femoral head were observed.RESULTS AND CONCLUSION: (1) The stress value of the acetabular cup was significantly higher than that of the acetabular body ((0.63± 0.34) vs. (1.89±0.67) MPa, P < 0.05). (2) The stress was concentrated on the femoral neck. The femoral head and femoral neck junction appeared with stress occlusion. The stress around the femoral prosthesis stem was (158.24±28.79) MPa, and the stress value of the femoral prosthesis stem was (186.46±22.37) MPa, showing no significant difference (P > 0.05). (3) The stress value showed significant difference among femoral head regions (P <0.05), and the stress of the regions 6 and 8 was significantly higher than that of the other regions (P < 0.05). (4) These results show that after metal-to-metal hip surface replacement, femoral neck stress is concentrated, the stress of the femoral prosthesis is occluded, and the stress at the medial femoral neck as well as the junction of femoral neck prosthesis and bone is larger.

Infection is a catastrophiccomplication in total knee arthroplasty.Reported a case of Nocardiafarcinica infection which appeared after application of cemented total knee replacement.A 64-year-old male patient was admitted to hospital 1 months after knee surgery,with a 2-week history of pain,swelling,and restricted mobility.As no improvement could be a-chieved after synovectomy and antibiotherapy,the prosthesis were removed from him.Although improvement could not be a-chieved in the knee of the patient at the end of 10-month therapy,the case has still being followed-up.

Human trophoblast cells were isolated and cultured in vitro in order to investigate possible pathogenesis of intrauterine infection caused by HCMV. Trophoblast cells were obtained by compound enzymes digestion and discontinuous percoll gradient. Cells and purity were identified by using immunocytochemistry assay with anti-CK7, Vim and beta-hCG antibodies. HCMV AD169 strain replication in isolated trophoblast cells and cell apoptosis were detected at different time points post infection (p.i.). The results showed that highly purified trophoblast cells were obtained. Specific virus replication was increased dramatically at the 24th h p.i., and then increased slowly during 48 h and 72 h. Apoptosis rate of trophoblast cells infected with HCMV was (34.68+/-3.14)% at 24th h p.i., while that in control group was (15.32+/-2.34)% (P<0.05). It was suggested that highly purified trophoblast cells can be isolated by the simplified cell purification method. HCMV can infect human trophoblast cells, and be quickly replicated, resulting in the accelerated apoptosis of human trophoblast cells during early time.

This study examined the impacts of intrauterine murine cytomegalovirus (MCMV) infection on the long-term learning and memory of offspring. Sexually matured male and female BALB/C mice without MCMV infection were identified by ELISA and then mated. Seventy pregnant mice were randomly divided into the virus group (n=40) and the control group (n=30), in which the pregnant mice were subjected to placenta inoculation of MCMV suspension (1 μL, 1×106 PFU) or the same amount of cell culture medium, respectively, at gestational age of 12.5 days. Some pregnant mice [virus group (n=20), control group (n=15)] were sacrificed by cervical dislocation at gestational age of 18.5 days, and the head circumference and brain weight of the mouse fetuses were measured, and the MCMV infection in their brain tissues was detected by PCR. The other pregnant mice [virus group (n=20), control group (n=15)] delivered naturally, and the learning and memory capability of the offspring at 70-day-old was analyzed by Morris water maze test. The results showed that 28.57% mouse fetuses in the virus group developed viral infection in the brain. Their head circumference and brain weight were significantly reduced as compared with those in the control group (P<0.01). The Morris water maze test revealed that the mouse offspring in the control group found the platform with straight-line trajectories after training. In contrast, the counterparts in the virus group intended to enter the central area, but looked for the platform with a circular trajectory. And the infected mice exhibited prolonged swimming distance and swimming latency (P<0.01). It was concluded that: (1) placenta inoculation of MCMV can cause fetal brain infection and intrauterine development retardation; (2) the offspring of MCMV placenta inoculation mice showed a long-term decline in learning and memory capability.

The present study examined von Willebrand factor (vWF) levels and ADAMTS13 activity in pregnant and severe preeclamptic women in order to shed light on the prothrombotic state in severe preeclampsia. Thirty healthy women of childbearing age, 22 second trimester pregnant women, 30 third trimester pregnant women and 10 severe preeclamptic patients were recruited in this study. ADAMTS13 activity was determined by the FRETS-vWF73 assay and vWF antigen (vWF:Ag) levels by an enzyme-linked immunosorbent assay. The results showed that there were statistically significant differences in plasma vWF antigen levels between the severe preeclamptic and third trimester pregnant women, between third and second trimester pregnant women (P<0.05). The third trimester pregnant women had significantly lower plasma ADAMTS13 activity than second trimester pregnant women (P<0.05). Nevertheless, no significant differences in plasma ADAMTS13 activity were found between severe preeclamptic patients and the third trimester pregnant women (P>0.05). In conclusion, plasma ADAMTS13 activity is normal in severe preeclampsia despite the increased vWF:Ag levels. Prothrombotic state is involved in the pathogenesis of severe preeclampsia, as a result of endothelial injury.

The effect of human cytomegalovirus (HCMV) on invasive capability of early pregnant extravillous cytotrophoblasts (EVTs) was investigated in vitro. Primary EVTs were obtained by complex phosphoesterasum digestion and gradient centrifugation from villous tissue aseptically taken from healthy pregnant women. Cytokeratin7 (CK7), vimentin (Vim) and c-erbB-2 were immunocytochemically detected to identify source of cells, and HCMVpp65 antigen was assayed to determine the infection state of primary EVTs by immunocytochemical staining. The EVTs were divided into two groups: control group and HCMV group, and the expression of c-erbB-2, matrix metalloproteinase-2 (MMP-2) and MMP-9 proteins was detected in two groups by immunocytochemistry and Western blotting. Enzymic activity changes of MMP-2 and MMP-9 were tested by gelatin zymography in primary EVTs infected with HCMV. The invasion of primary EVTs was detected by cell invasion assay in vitro after they were infected by HCMV. The cell source identification showed that the cells obtained were highly-pure primary EVTs, and primary EVTs could be infected by HCMV. Primary EVTs could express c-erbB-2, MMP-2 and MMP-9 proteins, and as compared with control group, the protein expression was decreased significantly in HCMV groups (P<0.05). Primary EVTs could secrete active MMP-2 and MMP-9 in vitro, and the activity of two MMPs was decreased significantly in HCMV groups (P<0.05). The in vitro cell invasion assay showed that the number of primary EVTs permeating Matrigel in HCMV group was decreased (P<0.05). We are led to conclude that HCMV can infect primary EVTs and inhibit their invasion capability, suggesting that the impaired EVT's invasion capability might be related to the abnormal expression of c-erbB-2, MMP-2 and MMP-9 proteins.

The present study examined von Willebrand factor (vWF) levels and ADAMTS13 activity in pregnant and severe preeclamptic women in order to shed light on the prothrombotic state in severe preeclampsia. Thirty healthy women of childbearing age, 22 second trimester pregnant women, 30 third trimester pregnant women and 10 severe preeclamptic patients were recruited in this study. ADAMTS13 activity was determined by the FRETS-vWF73 assay and vWF antigen (vWF:Ag) levels by an enzyme-linked immunosorbent assay. The results showed that there were statistically significant differences in plasma vWF antigen levels between the severe preeclamptic and third trimester pregnant women, between third and second trimester pregnant women (P<0.05). The third trimester pregnant women had significantly lower plasma ADAMTS13 activity than second trimester pregnant women (P<0.05). Nevertheless, no significant differences in plasma ADAMTS13 activity were found between severe preeclamptic patients and the third trimester pregnant women (P>0.05). In conclusion, plasma ADAMTS13 activity is normal in severe preeclampsia despite the increased vWF:Ag levels. Prothrombotic state is involved in the pathogenesis of severe preeclampsia, as a result of endothelial injury.
ADAM Proteins ; blood ; metabolism ; ADAMTS13 Protein ; Adult ; Blood Coagulation ; physiology ; Case-Control Studies ; Female ; Humans ; Pre-Eclampsia ; blood ; enzymology ; physiopathology ; Pregnancy ; Pregnancy Trimester, Third ; Young Adult ; von Willebrand Factor ; metabolism

Objective:To understand the current research status of master of professional degree students in clinical medicine in provincial teaching hospitals with moderate scientific research level under the " dual-track" training system.Methods:Sojump online survey was conducted to investigate the cognition, current situation and self-evaluation of all clinical master degree students in three grades in a provincial teaching hospital, and the research status of professional degree and scientific degree students was compared and analyzed respectively.Results:The proportion of scientific degree students participating in scientific research projects was significantly higher than that of professional degree students. The proportion of professional degree students participating in scientific research projects was still not high even in the third year of graduate students. However, there was no difference between scientific degree and professional degree students in the publication of scientific research papers. The scientific degree of scientific research knowledge is significantly higher than that of professional degree students. Although scientific degree students receive more scientific research guidance from their supervisors, professional degree students communicate more with their supervisors, and the results show that professional degree students are significantly more satisfied with their supervisors, graduate policies and scientific research policies than scientific degree students. In addition, the results of graduate students found that the degree of research stress of both scientific and professional degrees exceeded 50%.Conclusions:Scientific degree is better than professional degree in research status because of professional characteristics and more research guidance from their supervisors. However, through the reform of professional degree training mode based on scientific research project management in recent years, professional degree students have been able to communicate more with their supervisors, and their satisfaction with their supervisors and colleges has significantly increased. In addition, sufficient attention should be paid to high scientific research pressure of medical graduate students.