Objective To explore the value of high frequency and color doppler ultrasonography on diagnosis of deep venous thrombus in lower extremitiy.Method 32 cases with lower extremitiy deep venous thrombi diagnosed by angiography were further examined by CDFI,36 healthy cases were randomly selected as the normal group (NOR).Results High frequency and CDFI displayed the venous image in normal lower limbs rapidly and clearly as well as the echographic features of different shapes and types of the obstructed vessels.Conclusion High frequency and CDFI as a non-invasive and realtime method is better than angiography in diagnosis of the disease.

BACKGROUND:Nitric oxide can interfere with the function of mitochondria, and accelerate the intervertebral disc damage and degeneration by interfering with the release of inflammatory cytokines. Nitric oxide is an important inflammatory cel medium leading to degeneration of intervertebral disc induced by pressure and other external factors.
OBJECTIVE:To investigate the regulatory effect of nitric oxide and nitric oxide synthase inhibitor niacinamide on mitochondrial function and its association with biological behavior of rabbit nucleus pulposus.
METHODS:Cultured nucleus pulposus cel s of rabbit lumbar intervertebral disc were randomly divided into six groups:normal blank control group, 10μmol/L sodium nitroprusside group, 100μmol/L sodium nitroprusside group, 200μmol/L sodium nitroprusside group, 0.05 g/L nicotinamide group (100μmol/L sodium nitroprusside+0.05g/L nicotinamide), and 0.5 g/L nicotinamide group (100μmol/L sodium nitroprusside and 0.5 g/L nicotinamide). Different doses of nitric oxide donor sodium nitroprusside and nicotinamide were added in the medium of each group. Three days after intervention, cel proliferation activity, intracel ular ATP concentration, cel nitric oxide synthase activity, cel ular reactive oxygen species level, and mitochondrial membrane potential were detected respectively.
RESULTS AND CONCLUSION:(1) After 3 days of rabbit nucleus pulposus cel s intervened by different concentrations of sodium nitroprusside, intracel ular nitric oxide synthase content increased with sodium nitroprusside volume increase, and ATP concentration decreased along with sodium nitroprusside volume increase;there were significantly differences between the normal control group and sodium nitroprusside groups (P<0.01). (2) Reactive oxygen species could be increased in the sodium nitroprusside group. Niacinamide groups indicated a dose-dependent manner to improve the increase of cel ular reactive oxygen species levels with sodium nitroprusside intervention (P<0.01). (3) In the sodium nitroprusside groups, nucleus pulposus cel membrane potential decreased. In the niacinamide groups, sodium nitroprusside-induced decline in mitochondrial membrane potential was reduced (P<0.01). (4) Niacinamide groups also indicated a dose-dependent manner to improve the proliferative activity of nucleus pulposus cel s as compared with sodium nitroprusside groups (P<0.01). Significant differences were determined between the two groups (P<0.01). (5) Results suggest that the excess nitric oxide can damage mitochondrial metabolic function of rabbit nucleus pulposus cel s and cause cel energy metabolism. Niacinamide can reverse these damages by inhibiting nitric oxide synthesis, thereby contributing to the prevention against intervertebral disc degeneration.

Intervertebral disc (IVD) degeneration is considered to be the main cause of low back pain (LBP),however,there are lack of long-lasting and effective methods of clinical treatment.Tissue engineering technique based on stem cells becomes an essential research direction on repair of IVD degeneration at present,and its effectiveness and feasibility have been confirmed,but it is difficult to maintain the sufficiency and vitality of stem cells in IVD.Previous studies showed that stem cells existed naturally in IVD,and stem cells from stem cell niche could migrate to IVD physiologically to maintain the IVD environment balance under the adverse microenvironment.Unfortunately,these behaviors cannot preclude IVD degeneration.Therefore,theoretical basis for the regeneration of nucleus pulposus (NP) in situ can be obtained from studying the mechanism that the endogenous repair failure during IVD degeneration,the cell death and the migration of stem cells in IVD,and the key regulatory targets to sustain the quantity and quality of the stem cells.Although there have been few researches to study the mechanism of the cell death and the migration of stem cells in IVD so far,studies demonstrated that the major inducing factors of IVD degeneration (pressure and hypoxia) could decrease the number of NP cells by autophagy,apoptosis and necroptosis,and chemokines and their receptors played a critical role in the migration of mesenchymal stem cells.These researches provide a clue for studying the mechanism of endogenous repair failure during IVD degeneration.We reviewed the current research situation and progress of the mechanism that endogenous repair failure during IVD degeneration in the following articles.First,we exhibited the potential of IVD stem cells in IVD degeneration repair.Second,the effect of the adverse microenvironment (pressure,hypoxia,etc) on the migration of IVD stem cells was discussed.Third,the mechanism of the stem cell death,autophagy,apoptosis and necroptosis under the adverse (pressure,hypoxia,etc.) microenvironment,and the correlation between the IVD stem cells migration and autophagy,apoptosis and necroptosis was studied.And then tissue engineering of NP was also discussed to achieve the endogenous repair of IVD degeneration.These studies will provide an innovative research direction on endogenous repair and a new strategy of early therapy for IVD degeneration.

BACKGROUND:Intramedulary nail with rigid biomechanics was firstly selected for proximal femoral fracture. Proximal femoral intramedulary nail can effectively control the shortening and rotation of the fracture, alow patients to do early exercise and weight bearing, and promote the improvement of joint function. OBJECTIVE:To investigate the effects of proximal femoral intramedulary nail for treating intertrochanteric fractures on the recovery of joint function in young patients. METHODS:80 young patients with intertrochanteric fractures were randomly divided into the treatment group and the control group, with 40 cases in each group. The control group received dynamic hip screw fixation, and the treatment group received proximal femoral nail fixation. Perioperative index, complications and functional recovery of knee joint after 3 months of treatment were compared between the two groups. RESULTS AND CONCLUSION:Compared with the control group, length of incision, intraoperative blood loss, operative time, fracture healing time and postoperative ambulation time were significantly reduced in the treatment group (P< 0.05). Complications such as postoperative nonunion, hip varus, limb shortening and deep vein thrombosis were significantly less in the treatment group than in the control group (P < 0.05). After folow-up at 3 months post treatment, the excelent and good rate of hip function was significantly higher in the treatment group than in the control group (93%, 80%;P < 0.05). These results suggest that proximal femoral nail for treating intertrochanteric fractures in young patients folows the principle of biomechanics, reflects the characteristics of minimal invasion, and has few complications, and promotes the recovery of hip function.

Objective: To identify corticotropin releasing hormone(CRH) positive cells from the human placenta. Methods: Using immunohistochemistry to show the cellular expression of placental CRH. Results: During human pregnancy, placental CRH level rose markedly from undetectable amounts prior to 20 weeks gestation to nearly the peak level. Conclusion: The presence of CRH in maternal plasma is attributed to the placental production and subsequent release of this hormone into the maternal circulation. [

BACKGROUND: It has reported that nicotinamide is capable of protecting intervertebral disc (IVD) against interleukin-1β (IL-1β) or tumor necrosis factor-alpha (TNF-α) induced degeneration. However, the protective mechanism of nicotinamide on IVD cells apoptosis and proliferation remains unclear.OBJECTIVE: To investigate regulatory effects of nicotinamide on rabbit nucleus pulposus cell apoptosis and proliferation in vitro.DESIGN, TIME AND SETTING: Randomized control grouping design, which was carried out in the Laboratory of Orthopaedics and Stem Cell Center, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology from April to October 2007.MATERIALS: Ten Japanese white rabbits (aged 2-3 months weighing 1.5-2.0 kg) were used in this study. Furthermore, nucleus pulposus cells obtained from L1-6 lumbar spine were harvested and cultured for further experiments.METHODS: The NP cells were divided into 6 groups, including control group (without any drug as control), nicotinamide group (0.5 g/L nicotinamide), IL-1β group (10 μg/L IL-1β), IL-1β + caspase group (10 μg/L IL-1β and non-specific caspase inhibitor Z-VAD-FMK), IL-1β + small-dose nicotinamide group (10 μg/L IL-1β and 0.05 g/L nicotinamide), and IL-1β + large-dose nicotinamide group (10 μg/L IL-1β and 0.5 g/L nicotinamide). After 3 days of culture, the cells were examined with Annexin V-PI staining, caspase-3, 8 and 9 activity staining and MTT assay.MAIN OUTCOME MEASURES: The apoptotic rates, the positive rates of caspase-3, 8 and 9 activity staining and the absorbance of MTT assay of each group.RESULTS: ① As compared to IL-1β group, the apoptotic rates were decreased in the IL-1β + caspase group and IL-1β + large-dose nicotinamide group (P < 0.01). ②As compared to IL-1β group, the positive rates of caspase-3, 8 and 9 activity staining were decreased in the IL-1β + caspase group, IL-1β + large-dose and small-dose nicotinamide groups (P < 0.01 or P < 0.01). ③As compared to IL-1β group, the absorbance was increased in the IL-1β + caspase group and IL-1β + large-dose nicotinamide group (P < 0.01).CONCLUSION: Nicotinamide is capable of promoting cell proliferation and inhibiting IL-1β induced apoptosis of nucleus pulposus cells in vitro. The inhibition of apoptosis mainly acts via inhibition of the mitochondrial pathway.

Objective To generate an human interleukin-17 receptor-like molecule (IL-17RLM) recombinant plasmid with 6?myc tag and detect its specific expression in eukaryotic cells. Methods Design two specific primers(including the enzyme sites of EcoRⅠand XhoⅠ), reextract hIL-17RLM-L DNA fragment after PCR and insert it into the 6?myc tagged pcDNA3.0 vector, then detect its expression by Western blot after transfecting COS7 cells. Results The 6?myc tagged recombinant plasmid pcDNA3.0- 6?myc /hIL-17RLM-L was generated successfully and its expression can be detected by Western blot in eukaryotic cells. Conclusion The eukaryotic expressing plasmid pcDNA3.0-6?myc /hIL-17RLM-L was generated successfully and its specific expression was realized, which may provide the basis for further research of its biological function.

Objective To explore the effect of pulse irrigation combined with autologous blood transfusion on inflammatory markers and thrombosis index, such as C-reactive protein (CPR), erythrocyte sedimentation rate (ESR), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in patients undergoing total knee arthroplasty. Methods 96 patients due to knee osteoarthritis underwent unilateral total knee arthroplasty were randomly divided into experimental group (pulse irrigation combined with autologous blood transfusion after surgery ) and control group (alone autologous blood transfusion after surgery), PT, APTT, TNF-α, IL-6, D-Dimer of two groups at different time points were detected and compared. All patients underwent lower extremity vascular ultrasound examination after operation. Results PT, APTT values were no significant different between two groups (P > 0.05), TNF-α, IL-6 values of day1 and day7 after surgery in the experimental group were lower than the control group(P < 0.05), D-Dimer values of day4 after surgery were lower than the control group (P <0.05), the incidence of deep vein thrombosis (DVT) in the experimental group was lower than the control group (P < 0.05). Conclusion Pulse irrigation combined with postoperative autologous blood transfusion in total knee arthroplasty had no influence in blood coagulation , and can reduce inflammation and the incidence of DVT than alone autologous blood transfusion,thus is beneficial to the patient recover.

Objective To set up the optimized conditions,the amplification of eord blood CDCD+34 cells in vitro were analyzed by comparing the conditions such as different feeder-layers, stimulating-factors or purity/contents of those cells. Methods The cord blood CDCD+34 cells proliferation was analyzed by the methods of MTT, cell counting, and flow eytometer. The amplification and clone-forming ability of cord blood CDCD+34 cells were detected under optimized condition. Resulits The growth rates of cord blood CD+34 cell under optimized conditions(10 times) were significantly higher than that of the control(2.8 times) (P ＜0.01), and the cloneforming ability of cord blood CDCD+34 cells under optimized conditions(CFU-C 36.67±6.11) were also better than that of the control(CFU-C 16.33±1.53) (P ＜0.01). Conclusion The cord blood CD+34 cells proliferation can be promoted in the co-cultured system, and the character of the stem cells were kept well in that system.

To investigate the influence of recombinant adenovirus carrying tissue inhibitor of metalloproteinase-3 (RAdTIMP-3) on the main compositions of rabbits intervertebral discs and to assess its potential in treatment for intervertebral disc degeneration.[Method]RadTIMP-3 and empty adenovims vector with Lac-Z gene (Rad66) was propagated in 293 Cells and was purified, identified and tittered. Thirty Japanese white rabbits were randomly divided into 5 groups. And 25 μl of various reagents were injected to the L4、5 and L5、6 intervertebral discs of the rabbits as follows:normal saline in group 1, 1.0×1010 OPU/ml of RAd66 in Group 2, and 1.0×1010 OPU/ml of RAdTIMP-3 in group 3, 4 and 5. The intervertebral discs of each group were collected after 2, 2, 1, 2 and 4 weeks after injection respectively.Then X-gal staining, And Group 1, RT-PCR for TIMP-3 and aggrecan core protein,TUNEL staining, immunohistochemical staining for TIMP-3 and type I! Collagen and Safranin O-Fast green staining was carried out to assess the effects of RadTIMP-3 transfection.[Result](1)concentration of RAdTIMP-3 reached 1.9×1012 OPU/ml after propagation and purification. (2)RT-PCR shows that the expression of TIMP-3 was significantly raised in group 3, 4, 5, as compared with group 1 or 2. And the expression of core protein gene in group 3, 4, 5 increased slightly than in group 1 and 2. (3) TUNEL staining revealed that there was not significant difference between the positive-staining rates of any two of the groups. (4)TIMP-3 staining exhibited an obvious increase of positive-staining rates in group 3, 4 and 5 as compared with groupi or 2. The staining density of Safranin O-Fast Green staining and immunohistochemical staining for type II collagen of group 5 was obviously higher than that of group 1 or 2.[Conclusion]RAdTIMP-3 can express widely and safely in rabbit intervertebral discs, and improve the quantity and quality of matrix. It has the potential to be used in treatment for intervertabral disc degeneration.