In this study, microcalorimetry was adopted to establish a novel method for detecting the hemagglutination process of Radix Isatidis (Banlangen in Chinese, BLG), and to evaluate the hemagglutination activity diversity of BLG from various habitats. The hemagglutination biothermokinetics curves of positive reagent (phytohemagglutinin, PHA) and 8 batches BLG from different regions of the hemagglutination with 20% rabbit erythrocyte were recorded by microcalorimetry, then biothermokinetics parameters were abstracted, the hemagglutination utility of samples were calculated and analyzed by principal component analysis (PCA) and cluster analysis (CA), meanwhile the results were authenticated by micro-plate agglutination. It showed that the hemagglutination was an exothermic reaction, the reaction rate constant (k: 0.039-73.6 min(-1)), maximum reaction power (Pmax: -1 140.2 - 988.2 microW) and reaction enthalpy (Hi: -529.9 - 717.9 microJ) had good linear correlation with BLG extraction concentration (0.2-1.0 g mL(-1), r > 0.97), and PCA showed Pmax (531-1 335 microW) and Ht (585.2-989.2 microJ) could represent the hemagglutination activity diversity of BLG samples, just confirming with the results of micro-plate agglutination (the agglutination dilution was 3-11 respectively). According to the hemagglutination utility, the BLG samples from Good Agriculture Practice (GAP) regions, main producing area and general regions could be clustered correctly; meanwhile, the biothermokinetics curves with perfect distinctive fingerprint and specificity could give out more information for the quality control and evaluation for BLG. In conclusion, the microcalorimetry method established for detecting the hemagglutination activity of BLG samples on rabbit erythrocyte is sensitive and reliable, and could be adopted as an effective technique in detection aggulatination precisely, quantitatively and consecutively; and provide a novel approach for examining and evaluating quality for Chinese herbal medicine with aggulatinative activity such as BLG.

Objective To investigate the effect of minocycline on spinal CX3 C chemokine receptor 1(CX3 CR1)mRNA expression in morphine-tolerant rats with bone cancer pain.Methods Sixty female SD rats weighing 180-200 g in which intrathecal(IT)catheter was successfully placed at L3,4 interspace without complications were randomly divided into 4 groups:control group(group C,n=10);minocycline group(group M,n=10);bone cancer pain + morphine tolerance group(group BM,n=20)and bone cancer pain+morphine tolerance+ minocycline group(group BM+M,n=20).Bone cancer pain was induced by injection of breast cancer cells(Walker256)10μl(400/μl)into upper segment of bone marrow of right tibia.Morphine tolerance was induced by IT injection of morphine 20 μg/kg twice a day for 7 consecutive days starting from the 10th day after intratibia injection in BM and BM + M groups. Minocycline 0.25 mg/kg was injected IT once a day for 3 consecutive days in group M and after the model of bone cancer pain and morphine tolerance was established in group BM + M. Mechanical withdrawal threshold (MWT) and mechanical withdrawal duration (MWD) were determined before (T0, baseline) and at3, 6 and 9 days after operation (T1-3) and at 4, 7, 10 and 12 days after IT morphine injection was started (T4-7).The animals were sacrificed at T6 and T7 respectively in BM and BM + M groups and at T7 in C and M groups.The lumbar segment of the spinal cord (L4-6) was removed for determination of CX3 CR1 mRNA (by RT-PCR) and OX-42 expression (by immuno-histochemistry) .Results There was no significant difference in MWT and MWD at all time points between group C and group M. MWT was significantly decreased while MWD prolonged in morphine tolerant rats with cancer pain in group BM as compared with C and M groups. The hyperalgesia was significantly attenuated by IT minocycline in group BM + M. Spinal CX3 CR1 mRNA and OX-42 expression was significantly increased in group BM than in C and M groups. IT minocycline attenuated the increase in spinal CX3 CR, mRNA and OX-42 expression induced by bone cancer. Conclusion IT minocycline can inhibit spinal CX3CR1 mRNA expression, thereby antagonizing morphine tolerance in morphine-tolerant rats with bone cancer pain.

Objective To investigate the effects of intrathecal(IT)CX3 CR1 neutralizing antibody(antiFKR)on morphine tolerance in rats with bone cancer pain(BCP)and the unlerlying mechanism.Methods Forty-eight adult female SD rats aged 3 months weighing 180-200 g were randomized into 4 groups(n =12 each):group I sham operation(S); group Ⅱ BCP + normal saline(NS); group Ⅲ BCP + IgG(IgG)and group ⅣBCP + anti-FKR.Bone cancer pain(BCP)was induced by injecting Walker 256 cancer cells 10 μl(400 cells/ μl)into the medullary cavity of right tibia in groups Ⅱ,Ⅲ and Ⅳ.Ten days later morphine 20 μg/kg was administered IT twice a day for 7 consecutive days.Starting from the 8th day NS,IgG and anti-KFR 10 μl was administered IT once a day for 3 consecutive days in groups Ⅱ,Ⅲ and ⅣⅣ respectively.Paw withdrawal threshold to yon Frey filament stimulation(MWT)and paw withdrawal duration(MWD)were determined bcfore(To,baseline)and at 3,6,9 day after intra-tibial cancer cell inoculation(T1.2,3),on the 3rd and 7th day of IT morphine(T4.5)and on the 3rd day of IT NS/lgG/anti-KFR(T6).The animals were killed at T6 after last pain behavior assessment.The lumbar segment(L4-6)of the spinal cord was removed for determination of the expression of CX3 CR1 protein(by Western blot),μ-opioid receptor and TRPV1 receptor(by immuno-histochemistry)in the dorsal horn of spinal cord.Results IT morphine significantly eased BCP at T4,but morphine analgesia was significantly reduced on the 7th day of IT morphine in the 3 groups indicating morphine tolerance which was significantly relieved by anti-KFR in group Ⅳ.IT anti-KFR significantly down-regulated CX3CR1 prolein and TRPVI receptor expression and up-regulated μ-opioid receptor in group Ⅳ as compared with IT NS and lgG in groups Ⅱ and Ⅲ.Conctusion IT anti-KFR can relieve morphine tolerance in the rats with bone cancer pain by up-regulating μ-opioid receptor and down-regulating CX3 CR1 protein and TRPVI receptor expression.

Objective To evaluate the role of phosphatidylinositol 3-kinase (PI3K) p110β in spinal dorsal horn neurons in the development of arthritic pain (AP) in rats and the relationship with transient receptor potential vanilloid 1 (TRPV1) and acid-sensing ion channel (ASIC)1 a.Methods Forty adult female Sprague-Dawley rats in which intrathecal catheters were successfully placed,aged 3 months,weighing 250-300 g,were randomly divided into 4 groups (n =10 each):control group (group C),group AP,AP + PI3K p110β missense oligo-deoxynucleotide group (group MS) and AP + PI3K p110β antisense oligo-deoxynucleotide group (group AS).AP was induced by injecting complete Freund's adjuvant into the ankle joint cavity of right hindpaw.Normal saline 20 μl,missense oligo-deoxynucleotide 15 μg (20μl) and antisense oligo-deoxynucleotide 15 μg (20 μl) were administered intrathecally once a day for 6 consecutive days starting from the time immediately after arthritis was induced in groups AP,MS and AS,respectively.Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured 1 day before operation (T0) and on days 4,7,10 after operation (T1-3).The rats were then sacrificed after the last measurement of pain threshold at T3.L4-6 segment of the spinal cord was removed for detection of expression of PI3K p110β (by Western blot),and TRPV1 and ASICla (by immunohistochemistry)in spinal dorsal horn neurons.Results Compared with group C,MWT and TWL were significantly decreased atT1-3,and the expression of PI3K p110β,TRPV1 and ASIC1a was up-regulated in the other 3 groups(P＜ 0.01).MWT and TWL were significantly higher at T1-3,and the expression of PI3K p110β,TRPV1 and ASIC1a was lower in group AS than in groups AP and MS (P ＜ 0.01).Conclusion PI3K p110β in spinal dorsal horn neurons is involved in the development of AP in rats,and the mechanism is related to up-regulation of TRPV1 and ASIC1a expression in spinal dorsal horn neurons.

Objective To analyze the views and perceptions of practicing physicians on the practicing environment in the province,in order to provide policy reference for improvement.Methods By means of equi-probability and multi-stage sampling, 3 570 valid questionnaires were collected and analyzed from site survey which consist of the subj ective evaluation,and measures to identify and prevent medical professional risk.Results Overall assessment of the practice environment of physicians in Shanxi province is low,as those holding the environment asniceandacceptableaccounting for 4.8% and 21.7%,only 47.0% of physicians in tertiary hospitals hold the environment as very bad,while only 6.1% of those in level-1 hospitals share this view.92.4% of the physicians surveyed hold compliance of laws and regulations and technical specifications as key to prevention of medical dispute,yet most of them do not expect the media to be obj ective,fair and accurate in their coverage of events.Conclusions It is recommended to strengthen communication with the media for obj ective coverage of the limits of medicine,and explore the mechanism to take care of medical disputes by physicians,and improve their medical risk control capabilities,for a better practicing environment.

Objective To establish a rat model of bone cancer pain-chronic morphine tolerance. Methods Thirty-six adult female Sprague-Dawley rats weighing 180-200 g in which intrathecal catheters were successfully placed without complications were randomly divided into 3 groups ( n = 12 each) :group sham operation (group S),group chronic morphine tolerance (group M) and group bone cancer pain + chronic morphine tolerance (group BM). Bone cancer pain was induced by intra-tibia inoculation of Walker256 mammary gland carcinoma cells (4 ×102 cells/μl) in group BM, while in group M heat-inactivated Walker256 mammary gland carcinoma cells were given instead, and then 10 days later, intrathecal morphine 20 μg,/kg was administered twice a day for 9 consecutive days. The mechanical paw withdrawal threshold (MWT) and mechanical paw withdrawal duration (MWD) were measured before inoculation, at day 1, 3, 6 and 9 after inoculation, and at day 1, 3, 5, 7 and 9 of morphine administration. The degree of bone destruction was assessed by radiological analysis at day 9 after inoculation. After the last measurement of pain threshold, the rats were given innoxious touch-stimulus. The rats were sacrificed 3 h after stopping the stimulus, and L4-6 segment of the spinal cord was isolated to determine the expression of Fos protein in the spinal dorsal horn. Results Compared with group S, MWT was significantly decreased, MWD was significantly prolonged and the expression of Fos protein was up-regulated in group M ( P ＜ 0.05 or 0.01 ). MWT was significantly decreased, MWD was significantly prolonged, bone destruction scores were significantly increased,and the expression of Fos protein was up-regulated in group BM compared with group M ( P ＜ 0.05 or 0.01 ). Conclusion A rat model of bone cancer pain-chronic morphine tolerance is successfully established.

To establish a bioassay method and quality standard of Banlangen granula, agglutinated activity assay was used in the analysis of the traditional Chinese medicine, Banlangen granula. It showed that masculined effect could be picked up effectively and the products quality of different pharmaceutical factories and different batch numbers from the same factory could be revealed conveniently, accurately, quickly and directly with this method (valence value was between 2 and 11). The established bioassay method had a good reproducibility with RSD = 2%. The dependablity of the activity of red cell agglutination and restrainting influenza virus NA was conspicuous (r2 = 0.878 3). In conclusion, this bioassay method is suitable to control and evaluate the quality of Banlangen granula. Thus the method may provide a simple and effective technique in supervising and examining the quality of other traditional Chinese medicine.

ObjectiveTo investigate the correlation between any two of Capsule Endoscopy ScroringIndex (Lewis score),simplified Crohn Disease Activity Index (CDAI) and C-reactive protein (CRP) in small bowel Crohn disease (CD).MethodsA total of 58 consecutive patients with known small bowel CD were enrolled. We evaluated disease activity with Lewis score and simplified CDAI. Correlations among CRP,simplified CDAI and Lewis score were calculated with Spearman's rank order correlation coefficient.The optimal CRP cut-off value was calculated using the ROC curve.ResultsThe Lewis score showed inactive,mild and moderate-severe patients were 13,21 and 24,respectively.CRP of moderate-severe group was significantly higher than that in mild and inactive groups ( P ＜ 0.05 ).The optimal CRP cut-off value that differentiated patients with moderate to severe disease from the others was 13.50 mg/L with sensitivity of 87.5％ and specificity of 82.4％.The area under the ROC curve to analyze the cut-off was 0.849.Lewis score was moderately correlated with CRP (r =0.58,P ＜ 0.01 ),and weakly correlated with the simplified CDAI (r =0.40,P ＜ 0.01 ).ConclusionSerum CRP and the simplified CDAI cannot replace Lewis score for capsule endoscopy in the assessment of disease activity in small bowel CD.However,CRP may be considered as an inflammatory marker for evaluating the moderate to severe capsule endoscopic activity.

Objective To synthesize H.pylori bacterial ghosts (BG) and loaded with adriamycin.The cytotoxic effects in gastric cancer cell line were also observed.MethodsThe lysis plasmid was introduced into H.pylori by bacterial conjugation. H.pylori BG were produced by inducing H.pylori lysis at 42 ℃.After suspension and centrifuge, H.pylori BG were loaded with adriamycin.The adriamycin loading quantity was measured with spectrophotometry.The cytotoxic effects of H.pylori BG-adriamycin in gastric cancer cell line SGC7901 were evaluated with MTT assay.ResultsH.pylori BG were successfully synthesized and loaded with adriamycin.The loading quantity of adriamycin was 70.4 μg/mg.H.pylori BG were seen to be adsorbed and internalized by gastric cancer cells under confocal microscope, which distributed on the surface or cytoplasmic of SGC7901 cell line. Carried Adriamycin was delivered into gastric cancer cell line and mainly accumulated in the nucleus.IC50 of SGC7901 to H.pylori BG-adriamycin was 0.32 ± 0.15 by MTT assay, which was significantly lower than that to free adriamycin (0.44 ±0.15, P＜0.05).Conclusions The proliferation of gastric cancer cells were effectively inhibited by H.pylori BG-adriamycin.H.pylori BG are expected to be ideal carrier for anti-gastric cancer medicine.

Objective To evaluate the effect of intrathecal rapamycin on diabetic neuropathic pain in rats.Methods Thirty adult male Sprague-Dawley rats,weighing 180-220 g,in which IT catheters were successfully implanted,were randomly divided into 5 groups (n =6 each) using a random number table:control group (group C),diabetic neuropathic pain group (group DN),rapamycin 1 μg group (group R1),rapamycin 3 μg group (group R3) and rapamycin 10 μg (group R10).Diabetes mellitus was induced by intraperitoneal streptozotocin (STZ) 60 mg/kg on 5 days after IT catheters were implanted in DN,R1,R3 and R10 groups.Citric acid-sodium citrate buffer 6 ml/kg was injected intraperitoneally in group C.In R1,R3 and R10 groups,rapamycin (dissolved in 10 μl 4％ dimethyl sulfoxide) 1,3 and 10 μg were intrathecally injected,respectively,once a day for 7 consecutive days starting from day 21 after STZ injection,while the equal volume of 4％ dimethyl sulfoxide was given instead in C and DN groups.Paw withdrawal threshold (PWT) to yon Frey filament stimulation was measured before IT catheters were implanted,before STZ injection,on 7,14 and 21 days after STZ injection,and on 1,3,5 and 7 days after rapamycin administration.After measurement of PWT,the rats were sacrificed and L2-5 segments of the spinal cord were removed for determination of the expression of mTOR and phosphorylated mTOR (p-mTOR),S6K and phosphorylated S6K (p-S6K) (by Western blot) and expression of mTOR mRNA and S6K mRNA (by RT-PCR).Results Compared with group C,MWT was significantly decreased at 14 and 21 days after STZ injection in DN,R1,R3 and R10 groups,and the expression of mTOR,p-mTOR,S6K,p-S6K,mTOR mRNA and S6K mRNA was up-regulated in group DN (P ＜ 0.01).Compared with group DN,MWT was significantly increased at 5 and 7 days after rapamycin administration in group R1,at 3,5 and 7 days after rapamycin administration in group R3,and at 1,3,5 and 7 days after rapamycin administration in group R10,and the expression of mTOR,p-mTOR,S6K,p-S6K,mTOR mRNA and S6K mRNA was down-regulated in R1,R3 and R10 groups (P ＜ 0.01).Conclusion Intrathecal rapamycin can alleviate diabetic neuropathic pain in rats.