1.Therapeutic effect of anti-infective reconstituted bone xenograft on osteomyelit is in proximal tibia of rabbits
Zhi YUAN ; Yunyu HU ; Mingquan LI
Chinese Journal of Orthopaedics 1999;0(04):-
Objective To investigate the therapeutic effect of anti-infecti ve reconstituted bone xenograft(ARBX)on osteomyelitis. Methods A proximal tibia os teomyelitis rabbit model was used. Twenty animals were randomly assigned to 4 gr oups and all of them were injected with staphylococcus aureus through a bone win dow. Two weeks postoperatively the animals underwent clearance of the focal lesi on, followed by implant of 3 pellets of ARBX containing 30 mg of gentamicin in g roup Ⅰ, 3 pellets of reconstituted bone xenograft (RBX) in conjunction with 30 mg of intramuscular gentamicin for 5 days in group Ⅱ, 3 pellets of RBX without antibiotic in group Ⅲ, and those in group Ⅳ were left without bone grafting. S pecimens were harvested 8 weeks after the above procedures and were then subject ed to gross observation, radiological, histological and bacteriological examinat ions to compare their therapeutic effect on osteomyelitis. Results 1) In group Ⅰ the bacteria counting and modified Norden scoring were by far the smallest am ong all 4 groups (P
2.Bone inducing activity and antigenicity of native bone morphogenetic protein after various storage times
Hongxun SANG ; Dan LI ; Yunyu HU
Chinese Journal of Orthopaedics 1996;0(09):-
0.05). 3)ALP assay showed that the 7th,14th and 28th day of bBMP implantation, the ALP activities of bBMP in implantation groups were relatively higher than that of the control group (P0.05). 4)Histologically,after the 7th day implantation, a large quantity of mesenchymal cells differentiated with active chondrogenesis. The 14th day later, a large amount of cartilage and woven bone formed, and laminar bone and bone marrow were seen on day 28 after implantation. Conclusion The bBMP possesses active bone induction properties even after 10 years storage in 4℃ . The low-level antibodies of the recipient mice can be detected after implanted with bBMP, but the bone inducing activities are not affected. This may be a reference data for purification and clinical application of native BMP.
3.A study of nutritional assessment for the long term hospitalized elderly patients
Lu WANG ; Yunyu LI ; Wei JIANG ;
Chinese Journal of Geriatrics 2003;0(08):-
2 months were selected. Anthropometric measurements, laboratory examinations and dietary surveys were used to assess the nutritional status. ResultsBody weight(53.714.6)kg and body mass index (BMI) (19.34.6)kg/m~2were in low normal range. Total lymphocyte counts (2.000.71)10~9/L and serum transferrin contents (1.800.44)g/L were decreased and the creatinine-height index (42.4321.82)% was obviously lower than normal range of adult. ConclusionsThese patients have energy deficiency. The changes of the mentioned parameters could be the reflection of the nutritional status of these patients and also closely related to the changes of their body composition. It is necessary to evaluate nutritional state of the elderly patients. However, the characteristics of the body composition of the elderly patients should not be ignored and a global assessment is needed.
4.Clinical evaluation of nutrition status for 100 elderly patients with type 2 diabetes
Ziyan YANG ; Buo CHENG ; Yunyu LI ; Changping LI
Chinese Journal of Geriatrics 2001;0(05):-
Objective To evaluate the nutrition status of the elderly patients with type 2 diabetes. Methods The nutrition status of 100 elderly patients〔male 52, female 48, aged 60-83 yrs, with average of (69.2?0.7)yrs〕 with type 2 diabetes was investigated by analyzing nutrient content in daily diet, body measurement and laboratory examinations. Results Compared with the ideal body weight, total calorie intake was suitable for the elderly patients〔male: (1592?216) kCal, female: (1320?172) kCal〕, fat intake 〔male:(67?12) g, female:(65?13)g〕 was higher than the standard for diabetics(P
5.Comparison of risk of tumor invasion and metastasis under paravertebral block combined with general anesthesia versus general anesthesia in the patients undergoing radical lung cancer resection performed via video-assisted thoracoscope:plasma VEGF and M
Jiheng CHEN ; Zhiyi FAN ; Yunxiao ZHANG ; Yunyu JIN ; Ping LI
Chinese Journal of Anesthesiology 2015;35(1):23-26
Objective To compare the risk of tumor invasion and metastasis under paravertebral block (PVB) combined with general anesthesia versus general anesthesia in the patients undergoing radical resection for lung cancer performed via video-assisted thoracoscope in terms of plasma concentrations of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9).Methods Forty ASA physical status Ⅰ or Ⅱ patients of both sexes,aged 30-64 yr,with body mass index of 18-25 kg/m2,scheduled for elective radical resection for lung cancer performed via video-assisted thoracoscope,were randomly divided into 2 groups (n =20 each) using a random number table:general anesthesia group (group G) and PVB combined with general anesthesia (group PG).PVB of T4-7 was performed successfully with local injection of 0.375% ropivacaine 5 ml before induction of anesthesia.Double-lumen endotracheal tube was placed after induction of anesthesia,and the patients were mechanically ventilated.Anesthesia was maintained with inhalation of sevoflurane (end-tidal concentration 1%-2%),and intravenous infusion of remifentanil 0.2-0.3 μg · kg-1 · min-1,and intermittent intravenous boluses of atracurium.Before anesthesia and at 24 h after surgery,the venous blood samples were collected for measurement of plasma concentrations of VEGF and MMP-9.Results The plasma VEGF and MMP-9 concentrations were significantly lower after surgery in group PG than in group G.Conclusion PVB combined with general anesthesia significantly decreases the risk of tumor invasion and metastasis in the patients undergoing radical lung cancer resection performed via video-assisted thoracoscope in comparison to general anesthesia.
6.Expression and purification of rat brain NT-3 fusion protein and its antibody preparation
Zhiquan LI ; Yunyu HU ; Qingsheng ZHU ; Jinyu ZHU ; Fang LIU
Basic & Clinical Medicine 2006;0(03):-
Objective To clone NT-3 gene from normal rat brain and to purify its fusion protein and to prepare specific high titer antibody so that to provide a foundation for further study for peripheral nerve injury.MethodsWe amplified target gene by RT-PCR and cloned it into the vector of pMD-18T,then analyzed its sequence and compared it with the sequence from GenBank.We subcloned it into pRSET-A vector and introduced it into Escherichia coli BL21.The expression was induced by IPTG,and identified by SDS-PAGE.The fusion protein was purified by niccolum purify kit.We immuned rabbits with immunological adjuvant for specificity antibody preparation.Results We got a 777 bp gene segment by RT-PCR.The DNA sequence was identical to rat NT-3 gene sequence in GenBank.It proved that the target gene was correctly inserted into the vector.A new protein band of about 34 ku appeared on SDS-PAGE after induction of IPTG.A specific high titer antibody of 1∶64000 was gained by immunizing rabbits with adjuvant.
7.Mouse bone marrow derived mesenchymal stem cells suppress lymphocyte proliferation through co-Inhibitor B7-H1 upregulated by IFN-γ
Huiming SHENG ; Hongyu HUANG ; Yunyu WEI ; Ying WANG ; Li WANG ; Baihua SHEN ; Ningli LI
Chinese Journal of Microbiology and Immunology 2009;29(4):345-350
Objective To study the mechanism of mesenchymal stem cells immunosupression lym- phocyte proliferation via B7-H1/PD-1 pathway upregnlated by IFN-γ. Methods Bone marrow derived mes-enchymal stem cells (MSC) were isolated and purified by repeat adherent passage and detected them multi-potential differentiation in conditioned culture medium. Then MSC were cocuhured with lymphocyte prolifera-tion and assayed the level using γ H-thymidine incorporation. Meanwhile, ELISA measured IFN-γ, TGF-β, TNF-α and IL-10 in the cocuhured supernaatant and analyzed variation of B7-H1 molecular profile in MSC by flow cytometry. At last siRNA technology was deploied to interfere B7-H1 expression and analyzed MSC im-munosuppression on lymphocyte proliferation. Results In vitro the isolated MSC become homogeneous spi-die-shaped adherent cells after five passages, and in conditioned culture medium they could differentiate into adipocytes, osteocytes and chondrocytes. In the eocuhure of MSC with mixed lymphocyte, lymphocyte prolif- eration stimulated by Con A or by anti-CD3/CD28 antibody. The cpm value of the proliferation detected by 3H thymidine incorporation showed MSC suppressed the proliferation significantly (P = 0. 0167, 0. 0081,<0.0001 ) and the suppressive potential in a dose-dependent fashion. In the coeuhured supernatant cyto-kine IFN-T and TNF-α were detected in high concentration, but TGF-β, IL-10 were undetected. Simultane- ously MSC in the coeuhure upregulated B7-H1 expression from basic expression 7% to higher than 70% ( P < 0.05 ). After interfere B7-H1 expression in MSC by specific siRNA, we detected lymphocyte proliferation and got higher cpm by 3H thymidine incorporation ( P < 0. 05 ). Conclusion MSC upregulated B7-H1 mo-lecular expression upon the stimuli of IFN-γ, and through the B7-H1/PD-1 pathway mediated immunosu-pression on lymphocyte proliferation.
8.Longitudinal stress-strain relation of human peripheral vessels ex vivo
Qiang JIE ; Liu YANG ; Li ZHAO ; Yaotian HUANG ; Yunyu HU ; Jue LI
Chinese Journal of Tissue Engineering Research 2007;11(31):6318-6320
BACKGROUND: Due to the difference of species, the data of vessel in human are particularly useful for the clinical practice.OBJECTIVE: To analyze the longitudinal residue strain and the relationship between stress and strain of human limb arteries and veins, and explore the influence of different biomechanical properties on the repairs of limb injury.DESIGN: Observational trials.SETTING: Institute of Orthopaedics, Xijing Hospital of the Fourth Military Medical University of Chinese PLA.PARTICIPANTS: The experiment was carried out in the Xijing Hospital Affiliated to the Fourth Military Medical University of Chinese PLA from September 2005 to September 2006. The specimens were taken from 13 male amputee donors(who treated for accident injury), aged 18 to 30 years. Those tissue samples were used with the approval from the donors and offered by Xijing Hospital Affiliated to the Fourth Military Medical University of Chinese PLA.METHODS: ①Harvest and preservation of samples: The samples were obtained within 2 hours after death. The vessels were calibrated and harvested without any large branch to avoid the influence on the mechanical property of vessel wall,and then token on major vessels of limbs with Methylene Blue. The distance between the points token on vessel was measured by vernier caliper. The token vessels were cut and taken into Kreb's liquid in ice casement, then were kept into freezer (0-5 ℃). ②Longitudinal stretch ratio measurement: The vessels were taken into Kreb's liquid and the distance between the points token on vessel was measured by vernier caliper. The longitudinal residue strain was expressed by longitudinal stretch ratio. Lab temperature was 20-25 ℃, experiment was finished in 2 hours after sampling.③Stretch test: The vessel cut 1.0 cm was set into the instrument with Kreb's liquid for uniaxial tension test. The change length of each vascular specimen with or without the load and each load was measured three times and was averaged, lab temperature was 20-25 ℃, and experiments were finished in 5 hours after sampling. The curve of stress-strain was fitted by the measured data.MAIN OUTCOME MEASURES: Longitudinal stretching ratio, residue strain and stress-strain relationship of normal limb arteries and veins.RESULTS: The longitudinal stretch ratio of each artery decreased along vascular branch from proximal heart part to distal heart part, and that of each vein was contrast; There were significant difference in the longitudinal stretch ratios of major artery compared with those of saphena megna vein and branchiocephalicae vein (P < 0.001). The curve of artery shifting right showed the stiffness of vessels decreased along vascular branch from proximal heart part to distal heart part. That of vein shifting left showed the stiffness of vessels increased along vascular branch.CONCLUSION: With the major artery of human limbs from proximal end to distal end, both the longitudinal residue strain and the vascular stiffness gradually decreases, as for the vein, the condition is contrast. It suggests that the longitudinal biomechanical property should be involved into the consideration of repairing the artery and vein injuries of different sites.
10.Transforming growth factor beta as a regulator of the regional expression of the immune cytokines in xenogeneic bone transplantation
Hu XU ; Chunli ZHANG ; Mingquan LI ; Jianghao CHEN ; Yunyu HU ; Xinzhi XU
Chinese Journal of Tissue Engineering Research 2006;10(5):152-154
BACKGROUND: For xenogeneic bone transplantation, immune rejection is the major problem that affects the prognosis. However, the understanding about the expression and regulation of the immune factors in heterogenic bone transplantation is limited. Interleukin-1 (IL-1), interleukin-6 (IL-6)and tumor necrosis factor alpha (TNF-α) are important immune factors, and are closely related with post-transplantation rejection.OBJECTIVE: To observe the regional expressions of mRNA and protein of IL-1α, IL-6 and TNF-α in xenogeneic bone transplantation, and to in vestigate the effect of transforming growth factor (TGF)-β on these immune factors.DESIGN: A randomized controlled experiment. SETTING: The Orthopaedic Institute of Xijing Hospital of the Fourth Military Medical University of Chinese PLA MATERIALS: Totally 72 male Balb/c mice, with a body mass of 20 to 25 g, were randomly divided into 3 groups: combined bovine cancellous bone (bCB) granule group (Group A), simple bCB granule group (Group B) and blank control group (Group C) with 24 mice in each group.INTERVENTIONS: This experiment was conducted at the Institute of Orthopaedics, Xijing Hospital, Fourth Military Medical Univessity of Chinese PLA from June 2003 to June 2004. In Group A, one bCB combined TGF-β was implanted into the muscles of left thigh of each mouse. In Group B, one bCB alone was implanted, and in Group C, no bCB was implanted. The number of proliferation of cells in bone implantation area or adjacent tissues of the operated area was observed at postoperative 4, 7, 14 and 21 days; and the regional expressions of several immune factors in im plants were detected with in situ hybridization and immunocytochemistry methods.MAIN OUTCOME MEASURES: Histological observation and detection of regional cellular density of the implants of the mice in each group; the regional expressions of mRNA and protein of IL-1α,IL-6 and TNF-α in the implants RESULTS: ①) Histological observation and detection of regional cellular density of the implants of the mice in each group: on day 7, the cellular density of the proliferated tissues was significantly higher in the Group A than in the Group B [(470.63±132.89), (311.46±93.69)/field,P < 0.01];But on days 14 and 21, there was no significant difference. ②The regional expressions of mRNA and protein of IL-lα, IL-6 and TNF-α of the im plants of the mice in each group. On days 7 and 14 after xenografts were implanted, the regional expressions of IL-1α, IL-6 was respectively lower the xenografts combined with TGF-βthan in the simple xenogeneic bone (day 7: IL-1α 42.55±9.65 vs 67.95±17.82,IL-6 48.26±11.17 vs 77.21±15,16;day 14: IL-1α mRNA 84.77 ±7.42 vs 112.94±7.02,78.1 ±17.22 vs 121.18±15.44,P < 0.01) ,but for the TNF-α, there was no significant difference (P > 0.05).CONCLUSION: In the region of bone xenograft, a variety of cells express mRNAs and proteins of IL-1α, IL-6 and TNF-α, and their expressions are regulated by TGF-β. It may imply a kind of regulation of the immunity of bone xenograft by TGF-β.