Objective To explore the mechanism of long chain noncoding RNA RORin regulating prolifer-ation and apoptosis of pancreatic cancer cell. Methods Pancreatic cancer cell line BxPC-3 was selected. The RNA level of lncRNA ROR and notch1 was detected by RT-PCR.Notch1 protein level was detected by Western blot. The regulating relationship between lncRNA ROR and notch1 was analyzedby RNAhybird and luciferase re-porter assay. At last ,CCK-8 and TUNEL were applied to detectthe proliferation and apoptosis of cell line. Re-sults lncRNA ROR and notch1 were highly expressed in pancreatic cancer tissue ,compared with normal tis-sues. There was positive correlation between them. lncRNA ROR was over-expressed in BxPC-3,cell proliferation activity was increased and the percentagesof DNA damaged positive cells was decreased ,accompanied by in-creased levels of notch1 mRNA and protein. Luciferase assay confirmed that ROR could bind to notch1and inhibit its activity by miR-137. Compared with control group ,the proliferation of pcDNA-ROR + si-notch1 cells reduced and the proportion of TUNEL positive cells increased. The differences were statistically significant. Conclusionl ncRNA ROR regulated the proliferation and apoptosis of pancreatic cancer cells by promoting the expression of notch1.

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.

ObjectiveTo investigate the effect of operation treating spinal multi-level tuberculosis.Methods45 patients with multi-level spine tuberculosis were treated with debridement completely and anterior or lateral-anterior intervertebrae autograft. Of them, 5 patients added to anterior instrumentation.Results45 patients had been followed up for 12 to 40 months.The back pain of 40 cases were relieved within 5 weeks after operation, erythrocyte sedimentation rate decreased 3 weeks after operation. 38 patients who involved in kyphosis decreased their kyphosis mostly and only one patient lost 3 degrees in follow-up.15 patients who involved in neurological deficits improved one or two grades (Frankel). Grafts fused in 44 patients and there were no recurrent in follow-up.Conclusions Operative treatment is efficacious to multi-level body spine tuberculosis.

BACKGROUND:Recent studies have demonstrated that Niacinamide is capable of promoting the proliferation of intervertebral cells and improving intervertebral disc degeneration.Overloading is thought to the main cause of intervertebral disc degeneration.However,the protective effects of Niacinamide in loading induced intervertebral disc degeneration remains uncertain,OBJECTIVE:To investigate the protective effects of Niacinamide against axial loading induced degeneration of rabbit lumbar disc.DESIGN,TIME AND SETTING:A randomized controlled experiment was carded out in the Central Laboratory and the Laboratory of Department of Orthopaedics,Union Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology from November 2008 to April 2009.MATERIALS:Twenty-four Japanese white rabbits (4 months old,weighing 2.0 kg).Niacinamide was supplied by Tianjin Damao Chemical Reagent Factory.METHODS:Twenty-four Japanese white rabbits were randomly divided into 6 groups.The controllable axial loading induced rabbit lumbar disc degeneration model was adopted to impose 98N pressure on the rabbit discs to induce degeneration.Various doses of Niacinamide were given intragastrically to the rabbits in different groups:2 rabbits in group 1,the loading device was installed without pressing,and no Niacinamide was given;2 rabbits in group 2,given 50 mg/kg Niacinamide for 1 week;5 rabbits in group 3,loaded with 98N for 1 week;5 rabbits in group 4,loaded with 98N for 1 week,then the pressure was released for another week's recovery;5 rabbits in group 5,loaded with 98N and given 50 mg/kg Niacinamide for 1 week;5 rabbits in group 6,loaded with 98N for 1 week and then the pressure was released for another week's recovery,50 mg/kg Niacinamide was continually given during the 2 weeks.MAIN OUTCOME MEASURES:Magnetic resonance image and Thompson's grading system were used to assess degeneration degree of the discs;hematoxylin and eosin staining,immunohistochemical staining for type Ⅱ collagen,and Safranin O staining were used to evaluate histological changes;immunohistochemical staining for P161NK4A was used to evaluate cell proliferation and senescence.RESULTS：①According to the Thompson's grading system，there was no disc exhibited degeneration in group 2；5 rabbits graded Ⅱ in group 3；4 rabbits graded Ⅱ and 1 rabbit graded Ⅲ in group 4；2 rabbits graded Ⅰ and 3 rabbits graded Ⅱ in group 5；3 rabbits graded Ⅰ and 2 rabbits graded Ⅱ in group 6.MRI results revealed the alleviated degeneration in Niacinamide given groups.②The content of type Ⅱ collagen of annulus fibrosus of group 6 was 53.2% higher than that of group 4 (P＜0.01).③Safranin O-Fast Green staining density of group 2 was higher than that of group 1；The staining density of nucleus pulposus and annulus fibrosus of groups 5 and 6 was higher than the corresponding parts of group 4，especially that of nucleus pulposus (P＜0.01，P＜0.01)，and group 6 exhibited slightly increased levels than group 5.④P16INK4A positive staining rates decreased with the extension of Niacinamide administration time.CONCLUSION：Niacinamide can help to alleviate overloading-caused damage to intervertebral disc，and can benefit the recovery of damaged intervertebral disc.

OBJECTIVETo investigate the effect of Tiangui Gengnian soft capsule (TGC), which mainly consists of seabuckthorn fatty acid, on serum estrogen and estrogen receptor (ER) in multiple target tissues as uterus, liver and bone, in aged female rats, in order to explore its mechanism from the aspects of receptor and cytokines.

METHODSLow (0.72 g/kg), middle (1.80 g/kg) and high (4.50 g/kg) dose of TGC were administered by gastrogavage to young and aged (22 months old) female rats with osteoporosis for 45 days, and diethylstilbestrol (0.02 mg/kg) was used as a positive control. The levels of serum estradiol (E2), transforming growth factor beta1 (TGF-beta1), insulin-like growth factor-1 (IGF-1) were measured by radioimmunoassay and ELISA method, the protein expression of their receptor in bone, uterus and liver was detected by SABC immunohistochemistry, and the mRNA expression of E2 in uterus and liver detected by in situ hybridization with digoxin probe.

RESULTSIntervention of TGC could cause increase of serum E2, IGF-1 and TGF-beta1 levels, the TGF-beta1 reached 90.63 +/- 18.71 pg/L in the group administered with high dose, which was significantly different to that in the aged group (P < 0.01). There was no obvious effect of the mRNA expression of E2 in uterus and liver, and no effect of TGF-beta1 and IGF-1 in liver in rats.

CONCLUSIONTGC could improve the postmenopausal bone metabolism, alleviate and correct the bone loss, it is possibly realized by way of side-secreting/auto-secreting of E2 receptor and cytokines (TGF-beta1 and IGF-1) to improve the osteogenesis and inhibit the destruction of bone.

Animals ; Bone Density ; drug effects ; Bone and Bones ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Estradiol ; blood ; Female ; Insulin-Like Growth Factor Binding Protein 1 ; blood ; Osteoporosis ; drug therapy ; Phytotherapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Estrogen ; metabolism ; Transforming Growth Factor beta1 ; blood

Objective To investigate the role of negative-regulatory factors of toll-like receptors (TLRs)signal pathways in immunological pathogenesis of Kawasaki disease(KD).Methods Thirty-two chil- dren with Kawasaki disease and 16 age-matched healthy children were studied.Reverse-transcription PCR (RT-PCR)and real-time PCR were used to evaluate the mRNA expression levels of toll-like receptor 4(TLR4), MD-2,MyD88,IRAK-4,TRAF6,T1/ST2,IRAK-M,Triad 3A,and proinflammatory factors such as IL-1?, IL-6,IL-8 and TNF-?,in peripheral blood monocytes/macrophages(MC).The expression of TLR4 protein in MC was analyzed by flow cytometry.Results①Compared with the control group,the mRNA levels of TLR4, MD-2,MyD88,IRAK-4 and TRAF6 in KD group were up-regulated significantly(P＜0.01),and the expression level of TLR4 protein was also found to be up-regulated in KD group during acute phase.It was detected that expression levels of TLR4 protein in KD with coronary artery lesion(KD-CAL~+)was significantly higher than that of KD without coronary artery lesion(KD-CAL-)[flow cytometry:(6.5?1.7)% vs(11.9_+2.4)%,P＜0.01].②The expression level of negative-regulatory factors such as IRAK-M and Triad3A were significantly up-regulat- ed in acute phase of Kawasaki disease,while the mRNA levels of IRAK-M and Triad3A in KD-CAL~+ group was found to be significantly lower than those of KD-CAL~- group(P＜0.01).No difference of T1/ST2 mRNA expres sion level was detected among all groups(P＞0.05).③The expressions of proinflammatory eytokines such as IL-1?, IL-6,IL-8 and TNF-?in monoeytes/macrophages during acute phase of Kawasaki disease were higher than those of the control group(P＜0.01),and expression of proinflammatory cytokines in KD-CAL~+ group was significantly higher than that of KD-CAL~- group.Conclusion Relative insufficient expression of negative-regulatory factors, such as IRAK-M and Triad3A,maybe correlate with immunological pathogenesis of Kawasaki disease.

Objective To investigate the features of chronic prostatitis during puberty(CPP)and the effects of pelvic floor biofeedback therapy.Methods Totally,25 CPP children (mean age,16 years) and 15 children (mean age,16 years) with normal lower urinary tract as controls were included.In CPP group,NIH-CPSI scores were evaluated,expressed prostatic secretions (EPS) were examined,and bacterial culture was done;and CPP patients were categorized based on the definitions of NIH types.In both groups, urodynamic examination was performed,including evaluation of uroflow curve,maximum flow rate (Q_(max)), post-voiding residual urine (PVR),detrusor-sphincter dyssynergia (DSD),maximum detrusor pressure (P_(det,max))and maximum urethral closure pressure (MUCP).CPP patients underwent biofeedback therapy, and 10 weeks later the effects were assessed.Results In CPP group,NIH typing showedⅡ,ⅢA andⅢB in 1,3 and 21 cases,respectively.Before treatment in CPP and control groups,the incidence of staccato voiding (20 cases vs 1 case),DSD (22 cases vs 1 case),Q_(max)(10.7?3.7 vs 15.0?4.3ml/s),PVR (7.7?4.1vs 3.2?2.6ml),P_(det,max)(115.1?33.6vs 76.8?16.6cm H_2O)and MUCP(176.5?45.7 vs 86.2?28.5cm H_2O)all showed significant differences between the 2 groups(P＜0.05).In CPP group,the differences in pain(4.6?2.2 vs 2.1?1.6),urination (7.9?2.0vs 2.2?1.7),life impact (9.4?2.2vs 2.6?2.1)and total scores(22.0?5.2vs 7.0?4.2) of NIH-CPSI and Q_(max)(10.7?3.7 vs 14.9?5.6) between pre-and post-biofeedback were significant (P＜0.05).Conclusions The main type of CPP is categoryⅢB.The primary symptom is voiding disorder,which leads to greater psychological stress in patients.Children with CPP have pelvic floor dysfunctions and multiple abnormal urodynamic param- eters.The short-term effect of biofeedback strategies for CPP is satisfactory.

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.

BACKGROUND: Studies have reported that Niacinamide is capable of promoting the proliferation of intervertebral cell and protecting intervertebral disc (IVD) against interleukin-1 β-induced degeneration. However,the mechanism of Niacinamide underlying protecting IVD degeneration remains uncertain. OBJECTIVE: To investigate the regulatory effect of Niacinamide on interleukin-1 β-induced cell apoptosis and energy metabolism related gene in IVD in vitro. DESIGN, TIME AND SETTING: Experiments were performed in the Central Laboratory of Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology from November 2007 to May 2008. MATERIALS: Fifty-six IVDs from L16 lumbar spine often Japanese white rabbits,aged 3-4 months and weighing 1.5-2.0 kg,were harvested and cultured in alginate gel for further experiments. METHODS: IVD cultured models were randomly divided into 4 groups: normal control group (with the absence of drags), Niacinamide group (administrating 0.5 g/L Niacinamide), degeneration group (administrating 10 μg/L interlenkin-1β),treatment group (administrating 10 μg/L interleukin-1β and 0.5 g/L Niacinamide).After 2 weeks of culture,TUNEL staining and immunohistochcmical staining for FAS,Bcl-2, Caspase-3, hypoxia induced factor 1a, glucose transporter-1 and vascular endothelial cell growth factor were used to detect alternated cell apoptosis and expression of energy metabolism related genes. MAIN OUTCOME MEASURES: The positive cell rates of TUNEL staining and immunohistochemical staining in each group. RESULTS: The rate of TUNEL positive-staining cells of degeneration group was higher than normal control group (P=0.001). The rate of FAS positive-staining cells of degeneration group was obviously higher than normal control group (P＜0.01). The rate of Bcl-2 positive-staining cells of Niacinamide group was higher than normal control group (P=0.004). The rate of Caspase-3 positive-staining cells of treatment group was lower than degeneration group significantly (P=0,024).The rate of hypoxia induced factor- 1α positive-staining cells of Niacinamide group was lower than normal control group (P＜0.01),and the rate of degeneration group was higher than normal control group (P＜0.01 ). The rate of glucose transporter-1 positive-staining cells of treatment group was higher than normal control group(P＜0.01). CONCLUSION: Niacinamide can inhibit interleukin-1β-induced IVD cell apoptosis and alleviates interleukin-1β induced disturbance of energy metabolism.

BACKGROUND: The formation of bacterial biofilm on the material surface is the core problem of catheter-related urinary tract infection. Many researches have focused on the mechanism and prevention of such category of infection under static or simple hydrodynamic stimulation. The construction of dynamic model of bacterial biofilm of bladder urine flow close to real human diseases is the key to study the pathological mechanism and develop new technology of anti-biofilm infection. OBJECTIVE: To put forward the concept of turbulent flow shear stress of human bladder urine flow, construct this turbulent shear stress system based on the bacterial biofilm reactor of in vitro bionic human bladder, and explore the formation of E. coli biofilm stimulated by different stresses. METHODS: An in vitro dynamic bionic bladder urine flow model was designed. E. coli standard strain ATCC25922 was used as research object, and the medical silica gel was used as bacterial biofilm forming carrier. Four artificial urine flow stresses were simulated: hydrostatic pressure, constant turbulent flow shear stress, physiological turbulent flow shear stress and pathological turbulent flow shear stress (simulated urine retention environment). A bacterial biofilm reactor loaded with turbulent flow shear stress was established. Optical density value, colony count, and biofilm surface area of bacterial biofilm suspension were detected 24, 72, 120, and 168 hours. RESULTS AND CONCLUSION: (1) Optical density value of bacterial membrane suspension: there was significant difference between different urinary stress groups and different test time points (F=110.84, 187.96, all P < 0.000 1), and there was interaction effect between time and stress (F=50.05, P < 0.000 1). From hydrostatic pressure, constant turbulent flow shear stress, physiological turbulent flow shear stress, to pathological turbulent flow shear stress, the number of biofilm bacterial colonies increased. (2) Colony count of biofilm bacterial suspension smear: there was significant difference between different time (F=6.30, P=0.002 9); no difference was found between different urinary stress groups (F=1.11, P=0.400 1); and there was no interaction effect between time and stress (F=0.85, P=0.581 4). However, with the time extension of stress action, the colony count of complex stress group showed an increasing tendency, especially in the pathological turbulent shear stress. (3) Scanning electron microscopic characterization of biofilm bacteria: qualitative comparison between each group and different time points showed that the formation of bacterial biofilm was different from sparse fragments, lumps to large lumps. There were significant differences in the bacterial biofilm surface area between different urinary stress groups and at different times (F=505.72, 1 201.84, all P < 0.000 1), and there was interaction effect between time and stress (F=78.14, P < 0.000 1). From hydrostatic pressure, constant turbulent flow shear stress, physiological turbulent flow shear stress, to pathological turbulent flow shear stress, the biofilm formation increased significantly. (4) The results showed that this turbulent flow shear stress of human bladder urine flow can obviously stimulate E. coli biofilm formation in vitro. Its functional changes and pathogenic mechanism need to be further explored.