Objective To explore the mechanism and way for CT10 regulator of kinase like (CRKL) involving in drug resistance in leukemia cells. Methods The four major proteins included Ras protein, signal transducer and activator of transcripton 5 (STAT5) protein, phosphoinositide 3-kinase (PI3K) protein and paxillin protein in leukemia which involved in signal transduction pathway of CRKL. The expressions of those proteins were detected by Western-blot and immunofluorescent staining and confocal laser scanning microscopy. Results Compared with K562/S cells, the expressions of Ras(41.52±15.47 vs. 23.74±8.67) and PI3K (35.60±12.48 vs. 10.09±0.005) protein were up-regulated in K562/ADM cells (t=3.01,6.13;both P<0.05), while there were no significant changes in the expressions of paxillin (20.10±11.89 vs. 23.11±12.40) and STAT5 protein (25.72±14.46 vs. 17.58±9.21) between K562/S cells and K562/ADM cells(t=0. 18,1.43;both P>0. 0S). Conclusions Ras and PI3K protein may play a role in the multidrug resistance of K562 cell line, while paxillin and STAT5 protein may be not involved in the formation of resistant in K562 cells.

Objective The tissue engineering technique and the microsurgery technology is combined to construct the uncellular tissue engineering complex with vascularization and membrane guided dual effect. Through comparing study of using the simple biomembrane guided bone regeneration technique to construct the uncellular tissue engineering complex to repair the large segment bone defect in the animal body,the bone reparative effect of the tissue engineering bone wrapped by pedical fascial flap with vessels and that wrapped by the simple biomembrane was compared, thus to provide experimental evidence for the clinical application. Methods Twenty-four Newzland 5-month-old rabbits were used to build the bilateral periosteumincluded bone defect modelsin the middle piece of the ulna and the length of the defect was 1 cm. Autologous red bone marrow was implanted in the tissue engineering bone which was prepared by osteoinductive absorbing material including BMP. The prepared tissue engineering bone was implanted in the bone defect area. The right side was wrapped by the simple absorbable biomembrane, whereas the left side was wrapped by pedical fascial flap with blood supply. At the fourth, eighth, twelfth and sixteenth week after the operation each group was examined by the radiograph (x-ray), the light density measurement, gross morphology and histological inspection,bone shape measurement analysis in the repairing area and the biomechanics measurement at the twelfth week. The data was analyzed to test the difference of the bond defect repair. Results The radiograph, gross morphology and histological inspection showed the growth of vessels in the implant area, the quantity and the forming speed of the bone trabecula and, the cartilaginous tissue, the formation of the mature bone structure,remodeling of the diaphysis, recanalization of the cavum ossis and the absorption and the degradation of the implant of the group of pedical fascial flap with blood supply was superior to that of the group of the simple absorbable biomembrane. At the fourth, eighth, twelfth and sixteenth week after the operation the bone trabecula area were( 20. 35 ± 2. 41 ) %, ( 40. 21 ± 1.97 ) %, (66. 67 ± 3.44 ) % and ( 86. 47 ± 3.99) % respectively in the group of pedical fascial flap with blood supply, and were ( 7. 46 ± 2.64 ) %, ( 20. 66 ± 2. 28 ) % , ( 40. 22 ±1.84)% and(58. 18 ± 1.79) respectively in the group of the simple absorbable biomembrane. At the same time point after the operation the light density were 0. 636 ± 0. 012,0. 596 ± 0. 062,0. 552 ± 0. 009 and 0. 451 ±0. 008 respectively in the group of pedical fascial flap with blood supply, and 0. 742 ± 0. 032,0. 713 ± 0. 022,0. 655 ±0. 018 and 0. 606 ±0. 015 respectively in the group of the simple absorbable biomembrane. The units of blood vessel reproductive area in the bone repair junctional zone were ( 18.75 ± 2. 09 ) %, ( 37.41 ± 3.22 ) %,(53. 06 ±2. 18)% and (36.72 ±4. 73)% respectively in the group of pedical fascial flap with blood supply,and (5. 34 ± 1.17 ) %, (9. 48 ± 2.96) %, ( 22.43 ± 2. 21 ) % and ( 26. 27 ± 3. 14 ) % respectively in the group of the simple absorbable biomembrane. The biomechanics intension was 26.62 ± 3.96 in the group of pedical fascial flap with blood supply and 18. 38 ±0. 71 in the group of the simple absorbable biomembrane at the twelfth week after the operation. All of the differences were significant( P ＜0. 05 ). Conclusion The pedical fascial flap with blood supply has significant effect in promoting the tissue engineering bone to vascularize and promoting the bone formation by vascularization. The membrane guided bone regeneration technique restricted not only the growth of the fibrous connective tissue in the reparative process of the large segment bone defect effectively, but also the ability of fast and effective vascularization, thus the chronic creep and substitution process would be needed. Simple application of the biomembrane can compensate the shortcoming of chronic creep of the implanted material by the growth of the external callus.

BACKGROUND: Combined use of multiple interventions for different targets play superimposed or synergistic effects,which has become the current idea for spinal cord injury treatment.OBJECTIVE: To investigate the synergistic effects of low doses of 17-β estradiol combined with bone marrow mesenchymal stem cells (BMSCs) transplantation on the recovery of motor function and inflammatory reactions after spinal cord injury in rats.METHODS: The 10 of 70 male Sprague-Dawley rats served as sham group in which the spinal cord was only exposed but with no treatment, and the rest 60 rats were used to make animal models of spinal cord injury using modified Allen's method and then randomized into four groups (n=15 per group): model, estrogen, stem cell and combined treatment groups. Rats in the stem cell and combined treatment groups were given BMSCs transplantation at injured side; rats in the estrogen and combined treatment groups were given intramuscular injection of 17-β estradiol at 1 and 24 hours after modeling. At 1, 3, 5 and 7 days after modeling, rat functional recovery was evaluated by the Basso, Beatlie, Bresnahan score. The expressions of interleukin-1β and tumor necrosis factor-α in the injured spinal cord were detected by ELISA at 6, 12, 24, and 72 hours after modeling. Apoptosis in nerve cells was observed using TUNEL staining. RESULTS AND CONCLUSION: The Basso, Beatlie, Bresnahan scores were declined significantly after modeling,increased at 5 and 7 days after stem cell transplantation, estrogen treatment or their combined treatment (P < 0.05),especially in the combined treatment group (P < 0.05). The levels of interleukin-1β and tumor necrosis factor-α were elevated gradually after spinal cord injury (P < 0.05), but the levels decreased significantly at 12 and 24 hours in stem cell,estrogen and combined treatment groups (P < 0.05), and this decrease trend was more significant in the combined treatment group compared with the stem cell and estrogen groups (P < 0.05). At 72 hours after modeling, the rate of TUNEL positive cells was highest in the model group (P < 0.05) and lowest in the combined treatment group (P < 0.05).To conclude, the combined use of low doses of 17-β estradiol and BMSCs transplantation can facilitate the recovery of motor function after spinal cord injury by effectively inhibiting apoptosis in nerve cells.

Objective To investigate the role of CRKL activity in leukemia cells with muhidrug resistance and find new factor related to multidrug resistance. Methods By flow cytometry, CRKL activity was compared in K562, HL-60 cells and its resistance cells. The change of CRKL activity was observed in sensitive cells treated with and withdrawal daunorubicin. Results With the comparison of K562, HL-60 sensitive cells, in K562, HL-60 resistant cell lines, the level of CRKL phosphorylation in K562, HL-60 resistance cells treated with daunombicin 72 hours increased markedly. The level of CRKL phosphorylation was time-dependent with chemotherapy drugs, not change of CRKL activity was found in Jurkat ceils.Conclusion The level of CRKL activity is new factor related to muhidrug resistance in leukemia cells.

Objective: To study the relation among inducible nitric oxide synthase(iNOS), endothelial nitric oxide synthase(eNOS) and vascular endothelial growth factor (VEGF)in oral squamous cell carcinoma(OSCC) and their relation to clinical pathology and angiogenesis.Methods:OSCC tissue specimens were surgically obtained from 64 patients,the expression of iNOS,eNOS and VEGF in the samples were studied by immunohistochemistry technique. Microvessel density(MVD) was measured by counting the endothelial cells stained with anti-FⅧRAg antibody. Results: ①Positive VEGF and iNOS and eNOS immunostaining was detected in 39(60.94%),42(65.63%)and 45( 70.31%)of the cases respectively.②Expression of VEGF was significantly related to that of iNOS(P05).③MVD in OSCC tissue was positively correlated with the expression of VEGF and iNOS(P05).④VEGF expression was positively correlated to the pathological grade of OSCC(P0.05).Conclusion:iNOS,rather than eNOS, plays a positive role in OSCC development.iNOS and VEGF have a positive correlation in angiogenesis of OSCC.

Objective To investigate the inhibitory effect of hydroxy safflor yellow A ( HSYA ) on angiogenesis of H22 tumor-bearing mice and it's effects on the protein expression of MMP-3 .Methods After establishing the hep-atoma model for 24 h, the mice were randomly divided into control group , sorafenib group and HSYA group , the dose HSYA group received intraperitoneal injection at different dosages (1.125 and 2.25 mg/kg).The pathologi-cal changes were examined with HE staining , immunohistochemical staining and Western blot were applied to meas-ure the expression of angiogenesis related factor ( MMP-3 ) and we also detected the microvessel density with CD 34 . Results Compared with control group , the tumor cells proliferation and the new angiogenesis in HSYA group were suppressed .The expression of MMP-3 in HSYA group was significant reduced .Especially the dose of 2.25 mg/kg HSYA group ( P<0.01 ) , and tumor MVD-CD34 was also significantly reduced ( P<0.01 ) .But the effect is not better than sorafenib group .Conclusions HSYA may inhibit angiogenesis of tumor tissue in a certain concentration range and the anti-angiogenesis effect of HSYA may be related to inhibition of the protein expression of matrix met-alloproteinase-3 .

BACKGROUND:Studies have suggested that interleukin-6 is crucial for inducing cel apoptosis after acute spinal cord injury. OBJECTIVE:To observe the effect of interleukin-6 receptor monoclonal antibody combined with bone marrow mesenchymal stem cel s to treat acute spinal cord injury in rats. METHODS:Thirty Sprague-Dawley rats were randomly divided into sham group, model group (spinal cord injury group), treatment group 1 (interleukin-6 receptor monoclonal antibody transplantation group), treatment group 2 (bone marrow mesenchymal stem cel transplantation group), treatment group 3 (bone marrow mesenchymal stem cel+interleukin-6 receptor monoclonal antibody group), with six rats in each group. In the sham group, the spinal cord was only exposed with no injury, and in the other four groups, rat models of acute spinal cord injury were made using modified Al en’s method. Local injection treatment was performed in al the groups at 28 days after modeling. Basso, Beattie and Bresnahan (BBB) scoring and improved Tarlov scoring were used at 1 day before treatment and 1, 3, 7, 14, 28 days after treatment to test the hindlimb function. At 28 days after treatment, TUNEL method was used to detect cel apoptosis in the spinal cord. RESULTS AND CONCLUSION:Compared with the sham group, BBB scores and improved Tarlov scores were decreased significantly in the other four groups (P<0.05). At 7 days after treatment, the BBB scores and improved Tarlov scores in the treatment group 3 were significantly higher than those in the model group (P<0.05). At 14 days after treatment, the BBB scores and improved Tarlov scores in the treatment groups 1 and 2 were significantly higher than those in the model group (P<0.05);compared with the treatment group 2, the BBB score and improved Tarlov score were significantly increased in the treatment group 3 (P<0.05). Compared with the sham group, the number of apoptotic cel s was significantly increased in the other four groups (P<0.05);compared with the model group, the number of apoptotic cel s was significantly decreased in the three treatment groups (P<0.05);compared with the treatment group 2, the number of apoptotic cel s was significantly lower in the treatment group 3 (P<0.05). These findings indicate that the combined use of interleukin-6 receptor monoclonal antibody and bone marrow mesenchymal stem cel transplantation is better than bone marrow mesenchymal stem cel transplantation alone in the treatment of spinal cord injury, and interleukin-6 receptor monoclonal antibody reduces cel apoptosis in spinal cord injury, which is of positive significance for preventing against acute spinal cord injury.

Objective To discuss MRI diagnositic value and evaluation of the treatment of cervical Brucella spondylitis(BS).Meth-ods MRI data of 39 cases with cervical BS in our hospital were collected.Non-operative treatment,minimal invasive surgery and the open surgery were selected according to the MRI manifestations.The correlation was analyzed by consistensy check,imaging score and clinical effect evaluation.The data were analyzed by SPSS15.0.Results 39 cases of patients were examined by MRI before the treatment.37 cases were reviewed after 6 months treatment and 33 cases were reviewed after 12 months treatment.All of the Kappa values were more than 0.75 by imaging consistency check.MRI of cervical BS possessed characteristic expressions.The treatment effect according to MRI classification before the treatment:(1)drug therapy was performed in 14 cases (Group A);(2)minimal inva-sive surgery was performed in 7 cases (Group B);(3)focus clearance and bone graft were performed in 18 cases (Group C);(4)the clinical effects showed the healing rate in every group at different time point was different and the difference had statistical signifi-cance(P<0.05).The healing rate in Group C was higher than it in other group at 6 months after treatment.There was no significant difference among 3 groups or among different method group(Group C1,C2 and C3)in Group C at 12 months after treatment(P>0.05). Conclusion MRI classification for cervical BS has important value and guiding significance in the selection of clinical conservative treatment,minimal invasive surgery or the open surgery.

Objective To study the prevalence of retinopathy of prematurity (ROP) in preterm infants in Xiamen, Fujian. Design Retrospective case series. Participants From Oct. 2004 to Dec. 2008, 375 infants with birth weights ≤2500g or gestational age ≤32w were screened in Xiamen the First Hospital. Methods Initial examinations with the binocular indirect ophthalmoscopy and the scleral depression began at 4～6 weeks after birth or at 32 weeks postconception. All infants were followed-up until a stable retinal situation was reached. Main Outcome Measures The prevalence of ROP. Results Retinopathy of prematurity was detected in 10.93% of 375 neonates, including 18 eyes of stage 1, 48 eyes in stage 2, 14 eyes in stage 3, 2 eyes in AP-ROP. The prevalence of ROP in the infants with the birth weight ≤1000g, 1001-1500g and 1501-2000g was 45%, 17.07%, 7.01%, respectively; the occurrence rate of ROP in the infants with the gestational age ≤28 weeks, 29-30 weeks, 31-32 weeks, 33-34 weeks was 34.48%, 2.07%, 12.71%, 8.25%, respectively. Conclusions Earlier screening for ROP is very important. Low birth weight and young gestational age are the most important risk factors in the development of ROP.

BACKGROUND:Foreign scholars have obtained a success to cure fracture by implanting the complex of red bone marrow and formation factor.Due to the in vitro culture process is not necessary,the complex of red bona marrow and scaffold formation factor is only required to be implant immediately,called uncellular tissue engineered bone.OBJECTIVE:This study innovatively constructs uncellular tissue engineered bone with autologous red bone marrow wrapped by fascial flap with pedicle,and validates the superiority of repairing bone defects.DESIGN,TIME AND SETTING:Homebody controlled animal experiment was performed in the Hebei North University and the Experiment Center of the Affiliated Hospital to Hebei North University from December 2007 to February 2009.MATERIALS:A total of 24 News Zealand albino rabbits,aged 4-5 months; uncellular tissue engineered bone was a mixture of autologous red bone marrow and osteoinductive absorbing materials containing bone morphogenetic proteins.METHODS:Bone defect models were induced on adult New Zealand rabbits' right radial bone,left side served as control group,only implanted with osteoinductive absorbing materials complex,while right side served as experiment group,which contained fascial flap with pedicle.A fascial flap prepared with capillary network containing nameless blood vessel pedicle was located to be adjacent to the bone defect using micro-surgical technique,to wrap the tissue engineered bone and to fill the bone defect.MAIN OUTCOME MEASURES:At 4,8,12,16 weeks postoperation,six rabbits were tested by radiograph,spectrodensitometry,gross morphology observation,histological inspection,quantitative analysis of bone morphometry in bone defect area and analysis of vessels image in the junctional zone.RESULTS:①X-ray determination:At 16 weeks,the implant surrounding bone defects formed bone shaft structure in the control group,cortical bone was not continuous and medullary cavity was obstructed; in the experiment group,normal bone shaft structure was formed and recanalization of medullary cavity was observed.②Histological observation:At 16 weeks,few vessels grew into implant in the control group,mature bone trabecular were observed,and medullary cavity was obstructed; in the experiment group,the implanting materials were completely degraded and substituted by new bone,mature bone structure formed and recanalization of medullary cavity was observed.③Quantitative analysis of bone morphometry in bone defect area:At 4,8,12,16 weeks postoperation,the volume of bone trabecula in the experiment group was more than that in control group (P ＜ 0.05).④Analysis of vessels image in the junctional zone:The area of vessels in the unit area in the experiment group was greater than that in control group in every time stage (P＜0.05).CONCLUSION:The uncellular tissue engineering complex constructed by autologous red bone marrow wrapped by fascial flap with pedicle shows double effects of hymeno-inducing regeneration of bone and the vascularization.It is feasible to repair large-segment bone defects.It has obvious therapeutic effect in the aspects such as reducing the bone defect reparation time and advancing the quantity and quality of the bone generation.