Objective To explore the difference of gene profile between Cold syndrome and Heat syndrome in traditional Chinese medicine in stable and active stage of rheumatoid arthritis (RA). Methods Peripheral CD4+ lymphocytes from RA cases with defined Cold or Heat syndrome in TCM and healthy persons were obtained. Genechip analytical techniques were applied for analysis. Results There are 63 genes found different between active stage and stable stage of RA patients, and they refer to immune responses and signal transduction mainly. 48 genes were found different between Cold syndrome and Heat syndrome in stable stage of RA patients, and only 4 gene was found in the 63 genes for stable and active stage comparison, they are mainly referring to functional metabolism. 59 genes were found to be different among Cold syndrome and Heat syndrome in active stage of RA patients, and among them, no gene was same as in the 48 genes for Cold and Heat syndrome comparison in stable stage of RA, nor in the 63 genes for stable and active stage comparison. Conclusion The genes referred with Cold or Heat syndrome in the stable and active stage of RA are different with those genes referring to the difference of gene profile between active stage and active stage of RA, and further suggest that differentiation theory in traditional Chinese medicine have solid foundation in gene expression profile.

Objective To investigate the effects of Xuebijing injection on expression of myeloid cell triggering receptor-1(TREM-1)and the plasma levels of soluble TREM-1(sTREM-1),tumor necrosis factor-α(TNF-α), interleukin-6(IL-6)in patients with severe sepsis. Methods Twenty patients with severe sepsis admitted into Critical Care Medicine Department,Affiliated Hospital of Nantong University were given comprehensive treatments according to the guidelines for management of severe sepsis and septic shock(2004),and they were divided into Xuebijing group and control group(each 20 cases). The Xuebijing injection group was given Xuebijing injection 50 mL,3 times daily for 5-7 days followed by regular treatments. The changes in blood TREM-1 mRNA expression and plasma concentrations of sTREM-1,TNF-α,and IL-6 were detected before and after treatments on the 3rd and 5th day,and the above indexes were compared between the two groups. Results Before treatments,there were no significant differences in TREM-1 mRNA expression and levels of sTREM-1,TNF-α,IL-6 between two groups (all P>0.05). The TREM-1 expression and plasma concentrations of sTREM-1,TNF-α,and IL-6 of two groups were declined after treatments compared to their baselines,the degree of decline being more prominent in Xuebijing group〔TREM-1 mRNA 3 days:1.065±0.277 vs. 1.217±0.301,t=-3.267,P=0.047;5 days:0.912±0.239 vs. 1.071±0.254,t=-5.072,P=0.032;sTREM-1(ng/L):146.93±13.76 vs. 176.22±19.46,t=-5.442,P=0.033;TNF-α(ng/L):77.51±11.28 vs. 107.72±13.17,t=-4.355,P=0.032;IL-6(ng/L):288.35±14.59 vs. 323.89± 24.51, t=-3.941,P=0.028〕. Conclusion Early implication of Xuebijing injection is of great significance in patients with severe sepsis,it may reduce the expression level of TREM-1 and serum levels of downstream inflammatory mediators,that is beneficial to the control of inflammatory responses and improvement of systemic inflammatory response syndrome in such patients.

Objective To explore the value of procalcitonin (PCT) in the prediction of the prognosis and severity of bacterial infection in critically ill patients.Methods A total of 116 eligible patients with bacterial infection admitted in the intensive care unit were enrolled in this prospective study from February,2012 through November,2012.Within 24 hours after admission,the serum PCT was determined with immune-chromatography and acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score of patients was calculated.Based on the 28-day clinical outcome of patients,the patients were divided into fatal group (n =36) and survival group (n =80).The differences in PCT and APACHE Ⅱ score between the two groups were compared with t test or rank-sum test.The correlation between PCT and APACHE Ⅱ score was determined with Spearman's correlation analysis.Both PCT and APACHE Ⅱ score were analyzed separately and jointly with area under receiver operator characteristic curve (ROC curve,AUC) to predict 28-day survival.Comparison of prediction performance for predicting 28-day survival of patients with bacterial infection between PCT and APACHE Ⅱ was made with U test.Results PCT concentration was significantly higher in fatal group than that in survival group [5.36 (2.07,25) vs.0.24 (1.00,2.14)] (Z =5.596,P ＜0.01).APACHE Ⅱ score within 24 hour after admission was significantly higher in fatal group than that in survival group (24.30 ± 6.71) vs.(16.03 ± 7.23),t =6.147,P ＜ 0.01.Positive correlation between PCT and APACHE Ⅱscore was found to be statistical significance (r =0.388,P＜ 0.01).When rates of 28-day survival in patients were predicted by using PCT and APACHE Ⅱ score,the areas of under curve were 0.804 and 0.792,respectively.AUC of PCT was tenuously larger than that of APACHE Ⅱ score (U =0.2073,P =0.802).Using PCT and APACHE Ⅱ score together to predict 28-day survival,AUC (0.817) was increased.The joint prediction performance was higher than that of either alone,increasing the sensitivity to 90.7％ and the specificity to 75.2％.Conclusions Serum PCT can reflect the severity of the illness and prognosis of infectious disease in the intensive care unit.It can serve as a sensitive marker of predicting 28-day survival.Combining PCT and APACHE Ⅱ score together can increase the prognostic value.

Objective:To investigate the anti-inflammatory and analgesic effects of Rupixiao granules. Methods:Activity of Rupixi-ao granules against xylene-induced ear edema in mice was measured to evaluate the anti-inflammatory effect, while the analgesic effect was evaluated by acetic acid induced writhing test and hot plate test in mice. Results:Compared with the control group, Rupixiao gran-ules groups (1. 0, 2. 0 and 4. 0 g·kg·d-1) could significantly inhibit ear swelling in mice(P<0. 05 or 0. 001), and reduce the acetic acid induced writhing times(P<0. 01 or 0. 001). Rupixiao granules at medium and high dose could significantly increase the pain thresh-old of mice in the hot plate test. Conclusion:Rupixiao granules exhibit significant anti-inflammatory and analgesic activities.

Objective To study effects of Rupixiao granules on contraction of uterus of mice in vitro. Methods Mouse model with isolated uterine contraction was established. The mice pretreated with estradiol benzoate were sacrificed and then the uteri were taken out. The normal contractions were recorded by the biological function system. Effects of 10, 20, 30, 40, 50, 60 mg.mL-1 Rupixiao granules on spontaneous and oxytocin-induced (5 U.L-1) uterine contractions were recorded,inhibition of Rupixiao granules on mean constriction amplitude,constriction frequency and contraction activity of pitocin-pretreated uterus was observed and the inhibition rates were calculated. Results Compared with the baselines, the amplitude, frequency, activity of spontaneous uterine contraction were significantly down-regulated by Rupixiao solution at a range of 10-60 mg.mL-1 ,as well as it could inhibit oxytocin-induced spasmodic contraction of isolated uterus at 10-50 mg . mL-1 ( P<0. 01 ) . Conclusion The Rupixiao granules inhibits spontaneous contraction of isolated mouse uteri and oxytocin-induced spasm of uterine smooth muscle in vitro.

Objective To investigate the changes and clinical significance of the levels of matrix metallopro-teinase -9(MMP -9)in the serum and bronchoalveolar lavage fluid (BALF)in children with asthma whose had differ-ent levels of 25 -hydroxyl -vitamin D3 [25 (OH)D3 ]in the serum.Methods Fifty children with asthma between January 201 3 and January 201 5 were enrolled as the asthma group,based on the disease severity,and the patients were divided into the moderate to severe group (37 cases)and the mild group (1 3 cases),while 20 children with abnormal airway or tracheal foreign body were as a control group.The levels of 25 (OH)D3 and MMP -9 in the serum,levels of MMP -9 in BALF were measured and compared by using enzyme linked immunosorbent assay (ELISA)me-thod.Re-sults The level of serum 25 (OH)D3 in the asthma group[(1 9.5 ±6.4)μg/L]was lower than that in the control group[(39.3 ±7.1 )μg/L ],and there was a statistical difference between 2 groups(P <0.05).The level of serum 25(OH)D3 in moderate to severe group[(1 6.6 ±4.1 )μg/L]was lower than that in the mild group[(27.9 ±4.5)μg/L],there were statistically significant difference between 2 groups(P <0.05).There was no significant difference in the total number of cells and the percentage of macrophages among different serum 25(OH)D3 levels of children with asthma and the control group(P >0.05).The percentages of neutrophils,eosinophils and epithelial cells in BALF were significantly higher in different serum 25(OH)D3 levels of children with asthma than those of the control group(all P <0.05).There were significant difference levels of MMP -9 in the serum and BALF among different serum 25(OH)D3 levels of children with asthma and the control group(all P <0.05).In children with asthma,the levels of 25(OH)D3 in serum were significantly negatively correlated with the percentages of neutrophils,eosinophils and epithelial cells in BALF (r =-0.683,-0.795,-0.670,all P <0.05 ),exiting a significantly negatively correlation also seen between the serum levels of 25(OH)D3 and MMP -9 (r =-0.796,P <0.05).Conclusions Children with asthma often have low levels of serum 25 -(OH)D3 ,25(OH)D3 and MMP -9 may be involved in airway inflammation and airway remo-deling in children with asthma,and they may involve in the occurrence and development of asthma.

To confirm the etiology of a dead case for a 6 year-old female Ailurus fulgens,one strain of the predominant bacteria from pathologic tissues(heart,liver,spleen,lung and other samples) of the dead Ailurus fulgens were examined and isolated.The isolate was named R1 and no other bacteria were isolated.The bacterial etiological examination(morphological characteristics,biochemical characteristics and 16S rDNA gene detection)of R1 showed that it was identifed as K.peneumoniae.Artificial infection to mice about R1 was also conducted in this study.R1 had strong pathogenicity to mice and the LD50 is 6.5 × 104 CFU/mL.Moreover,the clinical and pathological features of the dead mice were consistent with that of the Ailurus fulgens.To find effective therapeutic drugs of curing other Ailurus fulgens,antibiotic sensitivity test of R1 was conducted,and the results revealed that R1 was highly sensitive to cefotaxime et al,moderately sensitive to amikacin and resistant to penicillin.These data showed that K.peneumoniae was bacterial pathogen leading to death of the Ailurus fulgens and it had strong resistance to penicillins,macrolides and virginiamycin and it had broad drug resistance spectrum.However,R1 is sensitive to cephalosporins and aminoglycoside antibiotics.

Objective To assess the clinical significance of detecting the immune markers in idiopathic thrombocytopenic purpura (ITP). Methods The frequencies of circulating B cells secreting platelet-specific antibody, platelet-specific antibody, the percentage of T lymphocyte subsets, the percentage of reticulated platelet and the level of thrombopoietin in 64 ITP patients and 31 healthy controls were measured with enzyme-linked immunospot assay (ELISPOT),modified monoclonal antibody immunobilization of platelet antigens assay (MAIPA), flow cytometry and sandwich enzyme-linked innnunosorbent assay respectively. Results Compared with the controls[1.3 ± 0. 5/105 peripheral blood mononuclear cell (PBMC), (0.33±0.06,0.41±0.03), (22.08±4.54)% and (8.19±2.46)%], the frequencies of circulating B cells secreting platelet-specific antibody (7.6±4.6/105 PBMC in acute ITP group, 5.3±3.0/105 PBMC in chronic ITP group), platelet-specific antibody (including the anti-GP Ⅱ b/Ⅲa antibody, anti-GP Ⅰ b/X antibody) (0.51 ±0.11, 0.48±0.06 in acute ITP group; 0.49±0.10,0.46±0.09 in chronic ITP group), the percentage of CD8+ T Lymphocyte (27.09±9.86 ) %, the percentage of reticulated platelet in ITP patients[the megakaryocyte cytosis group (24. 85 ± 19. 18)%, the normal megakaryocyte group (23.89±18.90)%]were significantly increased ( all P＜0.05).The frequencies of circulating B cells secreting platelet-specific antibody in acute ITP patients were notably increased (P＜0.05) compared to the chronic ITP patients. In T lymphocyte subsets, the percentage of CD3+T lymphocyte and CD4+ T lymphocyte and the ratio of CD4+/CD8+ in the patients with ITP[(60.88±14.59)%, (28.41±10.55)%, 1.18±0.59]were notably decreased than those in the healthy controls [(69.89±6.43)%, (35.38±5.05) %, 1.64±0.29, P＜0.05]. There was no apparent difference of the level of thrombopoietin between ITP patients with megakaryocyte cytosis (72. 09 ± 41.64 ) and health controls (75.37± 26. 32, P ＞ 0. 05 ), however, the level of thrombopoietin of ITP patients with normal megakaryocyte apparently increased (118.60±70.72, P＜0.05). Conclusion Detecting the frequencies of circulating B cells secreting platelet-specific antibody, platelet-specific antibody, the percentage of T lymphocyte subsets, the percentage of reticulated platelet and the level of thrombepoietin in the patients with ITP may improve the diagnosis and guide clinical therapy.

Objective: To establish a mouse hepatocellular carcinoma cell line that can produce mMIP-1α and to evaluate the possibility of cancer gene therapy by mMIP-1α. Methods: mMIP-1α cDNA was cloned into retrovirus vector pBabe puro and pBabe puro-mMIP-1α was constructed, then pBabe puro-mMIP-1α was used to transfect packaging cells, anti-puromycin cells was proliferated, the supernatant was used to infect hepa1-6, the anti-puromycin clone (hepa1-6 mMIP-1α) and hepa1-6 were analysed for the expression of mMIP-1α mRNA and protein by RT-PCR and immunohistochemistry respectively. The growth curve of hepa1-6 and hepa1-6 mMIP-1α was drawn. The chemotaxis of mMIP-1α produced by hepa1-6 mMIP-1α to mouse spleen cells was observed on agarose gel. C57B/L mouse was inoculated with the tumor cell and the tumorigenicity was studied. Results: Recombinant retrovirus vector pBabe puro-mMIP-1α with mMIP-1α cDNA was constructed. Hepa1-6 did not produce mMIP-1α mRNA and protein, while hepa1-6 mMIP-1α could produce mMIP-1α mRNA and protein. The growth curve of hepa1-6 and hepa1-6 mMIP-1α showed no difference. The chemotaxis of mMIP-1α produced by hepa1-6 mMIP-1α to mouse spleen cells was observed. The tumorigenicity was reduced. Conclusion: A mouse hepatocellular carcinoma Hepa1-6 mMIP-1α is established and mMIP-1α can affect the tumorigenecity of hepa1-6.

Objective: To establish a mouse hepatocellular carcinoma cell line that can produce mMIP 1? and to evaluate the possibility of cancer gene therapy by mMIP 1?. Methods: mMIP 1? cDNA was cloned into retrovirus vector pBabe puro and pBabe puro mMIP 1? was constructed, then pBabe puro mMIP 1? was used to transfect packaging cells, anti puromycin cells was proliferated, the supernatant was used to infect hepa1 6, the anti puromycin clone (hepa1 6 mMIP 1?) and hepa1 6 were analysed for the expression of mMIP 1? mRNA and protein by RT PCR and immunohistochemistry respectively. The growth curve of hepa1 6 and hepa1 6 mMIP 1? was drawn. The chemotaxis of mMIP 1? produced by hepa1 6 mMIP 1? to mouse spleen cells was observed on agarose gel. C57B/L mouse was inoculated with the tumor cell and the tumorigenicity was studied. Results: Recombinant retrovirus vector pBabe puro mMIP 1? with mMIP 1? cDNA was constructed. Hepa1 6 did not produce mMIP 1? mRNA and protein, while hepa1 6 mMIP 1? could produce mMIP 1? mRNA and protein. The growth curve of hepa1 6 and hepa1 6 mMIP 1? showed no difference. The chemotaxis of mMIP 1? produced by hepa1 6 mMIP 1? to mouse spleen cells was observed. The tumorigenicity was reduced. Conclusion: A mouse hepatocellular carcinoma Hepa1 6 mMIP 1? is established and mMIP 1? can affect the tumorigenecity of hepa1 6.