Objective To investigate the role of chemokine ligand 21 (CCL21) in the spinal cord in tibia bone cancer pain (BCP) in rats.Methods Forty adult female SD rats weighing 160-180 g were randomly divided into 5 groups (n=8 each):sham operation group (group Ⅰ ); sham operation + CCL21 neutralizing antibody group (groupⅡ); BCP group (group [); BCP + PBS group (group Ⅳ); BCP + control IgG group (groupⅤ)and BCP + CCL21 neutralizing antibody group (group Ⅵ).BCP was induced by inoculating Walker-256 mammary gland carcinoma cells into the rat tibia medullary cavity in groups Ⅲ-Ⅵ.PBS 15 μl,IgG 10 μg and CCL21 neutralizing antibody 10 μg were injected intrathecally (IT) at 14 days after intra-tibial injection of Walker-256 mammary gland cancer cells in groups Ⅳ- Ⅵ respectively.Mechanical withdrawal threshold to yon Frey filament stimulation (MWT) was measured at 1 day before (To,baseline) ; 7 and 14 d after Walker-256 cell injection (T1,T2)and at 0.5,1,2,4,8,12,24 and 48 h after intrathecal injection (T3-10 ).Results Intra-tibial injection of Walker-256 mammary gland cancer cells significantly decreased MWT as compared with the baseline values in administration of CCL21 neutralizing antibody at T5-8 as compared with MWT before intrathecal administration at T2 in group Ⅵ.MWT was significantly lower in groups Ⅲ- Ⅳ than in groups Ⅰ and Ⅱ.MWT was significantly higher at T5-8 in group Ⅵ than in groups Ⅲ - Ⅴ.Conclu]sion CCL21 in the spinal cord is involved in the maintenance of tibia BCP in rats.

Objective To evaluate the role of spinal IKK2/NF-κB pathway in the maintenance of bone cancer pain (BCP) in rats.Methods Twenty-eight unmated adult female Sprague-Dawley rats,weighing 160-200 g,were randomly divided into 4 groups (n =7 each) using a random number table:sham operation group (group S),BCP group (group BP),BCP + normal saline group (group BN),and BCP + BMS345541 group (group BB).BCP was induced by injecting Walker 256 mammary gland cancer cell suspension 5 μl (4 × 105 cells/ml) into the bone marrow of the right tibia of rats anesthetized with chloral hydrate in BP,BN and BB groups,while the equal volume of normal saline was injected in group S.On 10-12 days after operation,selective IKK2 inhibitor BMS345541 (50 μg/10 μl) was intrathecally injected once a day in group BB,and the equal volume of normal saline (10μl) was given once a day in group BN.The mechanical paw withdrawal threshold (MWT) was measured before intra-tibia injection (T0),on 7 days after intra-tibia injection (T1),at 1 h before drug administration and 1,2,4,12 and 24 h after drug administration on day 10 after operation,and at 4 h after drug administration on day 12 after operation (T2-8).The rats were sacrificed after MWT was measured at Ts and L4-6 segments of the spinal cord were removed for determination of phosphorylated NF-κB (p-NF-κB) expression (using Western blot analysis).Results Compared with group S,MWT was significantly decreased at T1-2,and the expression of p-NF-κB was up-regulated in BP,BN,and BB groups.Compared with group BP,MWT was significantly increased at T4-6,and the expression of p-NF-κB in the spinal cord was down-regulated in group BB.Conclusion Spinal IKK2/NF-κB signaling pathway is involved in the maintenance of bone cancer pain in rats.

Objective To investigate effects of intrathecal methotrexate on mechanical allodynia in rats with tibial bone cancer pain.Methods Forty-eight female SD rats weighing 150-180 g were randomly divided into 6 groups ( n =8 each):group Ⅰ sham operation + artificial cerebrospinal fluid(SA group),group Ⅱ sham operation + methotrexate 200 μg(SM group),group Ⅲ bone cancer pain + artificial cerebrospinal fluid(CA group),group Ⅳ-Ⅵ bone cancer pain + different doses of methotrexate (CM1-3 groups).The model of tibial bone cancer pain was induced by injecting Walker-256 cell into the tibial marrow cavity.CA and CM1-3 groups were intrathecal injected artificial cerebrospinal fluid,methotrexate 50,100 and 200 μg.SA and SM200 groups were intrathecal injected artificial cerebrospinal fluid and methotrexate 200 μg.The mechanical withdrawl threshold (MWT) was measured at day 1 before Walker-256 injection (baseline),7 day after injection (T0 ) and 2,4,8,24 hour and 1,3,5,7 days after intrathecal injection ( T1-8 ).Results Compered with the baseline,MWT was decrease in CA and CM1-s groups.Competed with To,MWT was decreased at T5-8 in CA group,MWT was increased at T3-5 in CM1 group,at T2-6 in CM2 group and at T2-7 in CM3 groups.MWT was decrease in CA and CM1-3 groups as compered with SA group; MWT was increased at T4-7 in CM1 group and at T3-7 in CM2 and CM3 groups.Conclusion Intrathecal injection of methotrexate can reduce tibial bone cancer pain in rats.

Objective: To explore the way of innovation and development of the pattern and means in nursing management in military hospital.Methods:Through various innovative practice from the following aspects,such as nursing service philosophy and measures for soldiers,the methods of nursing management and the elite-cultivating,nursing security management,which demonstrates the availability of innovation and development of the pattern and means in nursing management in military hospital.Results:The satisfactory rate of nursing service reached up to 99% in the examination of serving for soldiers of the whole army.The instruments which are manufactured for battle-scarred wounds and injuries use the flat of the Wound Nursing Centre,have reached the stage of clinical trial,we have completed five items of serial nursing research.The control rate of nursing quality of the whole hospital increased from 93.12% in 2003 to 98.9% in 2006,the satisfactory ratio of serious in-patient increased from 90.2% in 2003 to 96.7% in 2005.The ratio of health education achieves 100%.The satisfactory ratio of in-patient was 98.4% in September 2005,compared with 89.3% in February 2004.Conclusion:Only innovation can make nursing management develop and improve in military hospital.

Objective To investigate the association of serum C peptide concentration with the severity and the outcome of diabetic foot ulcer (DFU). Methods The clinical data of 257 inpatients with DFU were collected, including fasting and postprandial 2h C peptide levels and C peptide area under curve (AUCCP ). The patients were followed up on the outcomes of ulcers and death. The associations of serum C peptide concentration with the Wagner degree, infection severity, and healing rate were analyzed. Results The medians of fasting and 2h postprandial serum C peptide as well as AUCCP were 1. 37(0. 02 ~ 9. 00) nmol/ L, 3. 22(0. 02 ~ 29. 61) nmol/ L, and 511. 65 (3. 60 ~ 2 691. 30)nmol·min-1 ·L-1 respectively, which were lower than general levels. The time of follow-up in our study was 2. 8 (1. 0 ~ 5. 1) years. By the end of study, the wound of 75. 88% patients was healed, 3. 5%undergone major amputation, and 23. 74% died. After adjusting for relative factors, there were no significant associations of serum fasting and postprandial C peptide levels and AUCCP with Wagner degree and infection severity (P>0. 05). Cox regression analysis showed that the fasting plasma C peptide and hemoglobin were the independent protective factors for the healing of ulcers; old age, male, higher infection degree, and diabetes family history were their independent risk factors ( all P < 0. 05). Conclusions The lower plasma fasting C peptide concentration in patients with DFU is not correlated with Wagner degree and infection severity, but closely related with healing rate.

ObjectiveTo explore the effects of gabapentin on mechanical allodynia in rats with tibial bone cancer pain (BCP).MethodsForty-two female SD rats were randomized into 7 groups ( n=6):naive group (group N ),sham operation + NS control group (group SN),sham operation + GBP200mg/( kg · d) group (group SG200),BCP + NS control group (group BN),BCP + GBP50mg/( kg · d) group ( group BGS0),BCP +GBP100mg/(kg · d) group (group BG100),and BCP + GBP200mg/(kg · d) group (group BG200).The rats in group N,SN and BN received 5 ml normal saline and the rats in group SG200,BG50,BG100 and BG200 received 200,50,100 and 200 mg/( kg · d) dose of GBP via feeding from day 7 to 13 after operation,respectively.Mechanical withdrawal threshold(MWT) of the right paw and behavioral assays for ambulatory pain were measured just before operation and on days 1,3,5,7,8,10,12 and 14 after operation.ResultsMWT( (3.78 ± 0.38)g) in rats with BCP decreased and behavioral assays for ambulatory pain (0.76 ± 0.44) increased on day 7 after operation,as compared with those in group N ( ( 14.50 ± 1.38 ) g,(0.00 ± 0.00 ) ) and group sham ( ( 10.21 ± 0.88 ) g,( 0.00 ±0.00) ) (P ＜ 0.05 ).There was no apparent praxiological difference between group SN and group SG200 in a week of continuous application of gabapentin(P＞ 0.05 ).Compared with those in group BN,there was no change on MWT in group BG50 (P ＞ 0.05 ),and however,behavioral assays for ambulatory pain decreased (P ＜ 0.05 ).MWT in group BG100( (5.35 ±0.85)g) and BG200( (5.71 ±0.72) g) increased in day 10 after operation,as compared with those in group BN ( ( 2.61 ± 0.40) g) and group BG50 ( ( 3.28 ± 1.15 ) g) (P ＜ 0.05 ),and the difference was still statistically significant until day 14 (P ＜ 0.05 ).Behavioral assays for ambulatory pain in group BG100 and BG200 decreased from day 8 after operation,as compared with those in group BN and group BG50 (P ＜ 0.05 ).ConclusionGabapentin,in medium to large dosage,can inhibit pain reaction of rats with bone cancer pain.Nevertheless,with the development of cancer,the effect of gabapentin decreases.

Objective To evaluate the effects of intrathecal methotrexate on the activation of microglia in spinal cord in a rat model of tibial cancer pain (TCP).Methods Thirty female Sprague-Dawley rats,aged 5-7 weeks,weighing 150-180 g,were randomly divided into 3 groups (n =10 each):sham operation + artificial cerebrospinal fluid (group SA),TCP + artificial cerebrospinal fluid (group CA),and TCP + methotrexate (group CM).TCP was induced by injecting Walker-256 cancer cells into the medullary cavity of tibia.Artificial cerebrospinal fluid or methotrexate 100μg (15μl) was injected intrathecally over 10 min on 7th day after TCP.Mechanical pain threshold (MPT) was measured before TCP,at 1,3,5 and 7 days after TCP and 2,4,8 and 24 h after administration (T0-8).The rats were sacrificed after measurement of the pain threshold at T8 and the spinal cord was isolated for detection of the activation of microglia (by immunofluorescence) and content of tumor necrosis factor-α (TNF-α) and IL-1β (by ELISA).Results Compared with group SA,MPT was significantly decreased,and the number of activated microglia cells in the spinal cord was increased,and the contents of TNF-α and IL-1β were increased in groups CA and CM (P ＜ 0.05).Compared with group CA,MPT was significantly increased,and the number of activated microglia cells in the spinal cord was decreased,and the contents of TNF-α and IL-1β were decreased in group CM (P ＜ 0.05).Conclusion The mechanism by which intrathecal methotrexate reduces TCP in rats is related to inhibition of the activation of microglia and reduction of the secretion of proinflammatory cytokines TNF-α and IL-1β in the spinal cord.

Objective To evaluate the role of chemokine receptor CXCR4 in spinal dorsal horn in the development of morphine tolerance in rats with bone cancer pain (BCP).Methods Forty female Sprague-Dawley rats,weighing 180-220 g,were equally randomized into 5 groups using a random number table:sham operation group (group S),BCP group (group B),BCP + AMD3100 (specific CXCR4 antagonist) group (group BA),BCP + morphine group (group BM),BCP + morphine + AMD3100 group (group BMA).BCP was induced by injecting Walker 256 mammary gland cancer cell suspension (4 × 105 cells/ml) 5 μl into the bone marrow of the right tibia of rats anesthetized with chloral hydrate.On 6 days after injection of mammary gland cancer cells,AMD3100 2 mg/kg was intraperitoneally injected twice a day for 9 days in BA group,and morphine 10 mg/kg was subcutaneously injected twice a day for 9 days in BM group.AMD3100 was intraperitoneally injected and morphine was subcutaneously injected as previously described at the corresponding time point in BMA group.Before injection of mammary gland cancer cells (T0) and on 4,6,8,10,12 and 14 days after injection of mammary gland cancer cells (T1-6),paw withdrawal threshold to von Frey hair mechanical stimulation (PWMT) was measured.The rats were then sacrificed and L3-5 segments of the spinal cord were removed for determination of c-fos expression in spinal dorsal horn using immunofluorescence.Results Compared with S group,PMWT was significantly decreased at T2-6 in B and BA groups and at T4-6 in BM group,and c-fos expression was up-regulated at T6 in BM group (P ＜0.01).PMWT was significantly higher at T3-5 in BM group and at T3-6 in BMA group than in group B (P ＜ 0.01).Compared with BM group,PMWT was significantly increased at T5,6 and c-fos expression was down-regulated at T6 in BMA group (P ＜ 0.01).Conclusion Chemokine receptor CXCR4 in spinal dorsal horn is involved in the development of morphine tolerance in rats with BCP and the mechanism may be related to activation of c-fos.

Objective To evaluate the role of chemokine CXCL12 in the spinal cord in the development of bone cancer pain (BCP) in rats and the relationship with microglial activation.Methods Thirty-two female Sprague-Dawley rats,weighing 180-220 g,were equally randomized into 4 groups (n =8 each) using a random number table:sham operation group (group S),BCP group (group B),BCP + CXCL12 neutralizing antibody group (group BC),and BCP + IgG control antibody group (group BI).BCP was induced by injecting Walker 256 mammary gland cancer cell suspension (4 × 105 cells/ml) 5 μl into the bone marrow of the right tibia of rats anesthetized with chloral hydrate in B,BC and BI groups,while the equal volume of normal saline was injected instead in group S.On 12,13 and 14 days after injection of mammary gland cancer cells,CXCL12 neutralizing antibody 10 μg/15 μl was intrathecally injected once a day in group BC,while IgG control antibody 10 μg/15 μl was intrathecally injected once a day in group BI.Before injection of mammary gland cancer cells (T0) and on 3,5,7,10,12 and 14 days after injection of mammary gland cancer cells (T16),paw withdrawal threshold to mechanical stimulation (PWMT) was measured.The rats were then sacrificed and L4,5 segments of the spinal cord were removed for determination of Iba-1 (pan-microglial marker) expression in spinal dorsal horn using immunofluorescence after PWMT measurement at T6.Results Compared with S group,PMWT was significantly decreased at T2-6,and Iba-1 expression was up-regulated at T6 in B,BC and BI groups (P ＜ 0.01).Compared with B group,PMWT was significantly increased at T5,6 and Iba-1 expression was down-regulated at T6 in BC group (P ＜ 0.01).Conclusion Chemokine CXCL12 in the spinal cord is involved in the development of BCP,and microglial activation is involved in the mechanism.

Objective To explore the distribution of pathogenic micro-organisms in the diabetic patients with foot infection,and to investigate the characteristics of the pathogen distribution and its relationship with the severity of diabetic foot ulcer( DFU ).Methods Ninety diabetic foot patients with positive culture of micro-organisms were classified into Wagner 1-5 grades according to the severity of DFU,which was ranked as mild,moderate,and severe infection based on the gradient of infection.The family,genus,and strain of micro-organisms were analyzed,and associated factors were discussed in regard to the kinds and severity of DFU and infection.Results With rising Wagner's grades and aggravating infection,the bacterial floras were transformed from gram-positive cocci( 61.1％ in Wagner 1 ) to gram-negative rods( 80.0％ in Wagner 5),at the same time,composite( 10.0％ to 28.6％ ) and opportunist ( 83.3％ to 100％ ) infections were increased.Wagner's grade,severity of infection,previous treatment,patients' age,control of blood glucose,course of DFU,etc,were all related to types of infection and opportunist infection ( P＜0.05 ).Conclusions Proportion of gram-negative bacilli with opportunist composite infection seems to increase in patients with severe DFU and severe infection requiring appropriate application of antibiotics.