Objective To explore the correlation between the change of intestinal flora and insulin resistance in pre-diabetes population,and the role of intestinal flora in the occurrence and development in the pre-diabetes population.Methods two hundreds and fifty cases of pre-diabetes in our hospital were selected and divided into the group A(pre-diabetes intervention group)and B(prediabetes non-treatment group).Fifty cases of diabetes(positive control group)served as the group C.The cohort study was adopted.The follow-up intervention lasted for two years.The intestinal flora and blood biochemical indicators were detected in different groups and at different time periods.The insulin resistance index was calculated.Results The total bacterial count before intervention in the group A and B was significantly higher than that in the group C;Enterococcus genus in the group C was significantly higher than that in the group A and B;the proteus and lactobacillus content in the group A and B was higher than that in the group C,but the difference was not significant;Bifidobacterium genus in the group C was higher than that in the group A and B.After intervention,the total bacterial amount in the group Awas significantly higher than the group B and C,but there was no statistical difference between the group B and C;enterococcus genus in the group C was significantly higher than that in the group A and B,enterococcus genus after intervention in the group A was reduced and which in the group B was increased,the difference had statistical significance;the proteus and lactobacillus content in the group A and B was higher than that in the group C,but the difference was not significant;bifidobacterium genus in the group C was higher than that in the group A and B,moreover which after intervention in the group B was increased,the difference was statistically significant.The insulin resistance before processing had no statistical difference between the groups A and B,which in the group C was higher than the group A and B,the insulin resistance after intervention in the group A was decreased,the difference was statistically significant.The total bacterial count,enterococcus and bifidobacterium were positively correlated with insulin resistance,and bacteroides,proteus and lactobacillus had no significant correlation with insulin resistance.Conclusion Intestinal flora has correlation with insulin resistance,and serves as a marker of pre-diabetes development,implementing the probiotics adjustment combined the quantization aerobic muscular movement can realize the adjustment of insulin sensitivity and improves the diabetic development in the prediabetic population.

Objective To study the potential role of PKC inhibitor Rottlerin in the treatment of asthma. Methods Balb/c mice were sensitized and challenged with ovalbumin(OVA) protein to construct murine experimental model of asthma. Then the mice were treated by means of injection of Rottlerin or phosphate buffered saline (PBS) into the abdominal cavity. Changes of the total serum IgE, pulmonary eosinophils, cytokines and pulmonary inflammation were investigated. Results Injection of Rottlerin at the dose 0.3 mg/kg into the murine abdominal cavity could inhibit the infiltration of pulmonary eosinophils and pulmonary inflammation and significantly decrease the production of total serum IgE and Th 2 cytokines as well. Conclusion Rottlerin can inhibit murine experimental asthma.

This study was aimed to establish the TLC identification method of Radix Mirab ilis himalaic a. The β-sitosterol and daucosterol were used as the reference substances. The single-factor test was used. A variety of factors which affected TLC were systematically investigated to filter out the best TLC conditions for identification of different batches of medicines. The results showed that the best TLC conditions were as follows: silica gel G plates, extraction solvent (methanol), reagent (5% sulfuric acid in ethanol), extraction method (ultrasonic extraction with methanol), ex-tracted time (30 min), the agent (petroleum ether-ethyl acetate-acetone (5:2:1)) and sample volume (6 μL). It was concluded that the method, which had high separation degree, was reproducible and simple. It can be used as the quality control of Radix Mirab ilis himalaic a.

Objective To compare differences and similarities of the content of trigonelline in Himalaica mirabilis of either wild or cultivated materials from different places. Methods An HPLC method was established to determine the content of trigonelline in Himalaica mirabilis. ZORBAX XDB-CN column (4.6 mm×250 mm, 5 μm) was used, with mobile phase of acetonitrile-0.03%acetic acid (85∶15), flow rate of 0.8 mL/min, detection wavelength of 265 nm, determination wavelength of 360 nm, and column temperature of 30 ℃. Results Regression calculation was made on peak area with the reference solution concentration, and then got the regression equation A=23.409C-26.398, r=0.999 8. Trigonelline showed good linear relation with peak area among the range of 2.004-200.400 μg/mL. The average recovery of trigonelline was 99.57%, RSD=1.11%. Conclusion There was no significant difference in the content of trigonelline of either wild or cultivated materials from different places. This study laid the foundation of application of the cultivated Himalaica mirabilis.

Objective To investigate the distribution of congenital cardiovascular malformations in fetuses with chromosomal abnormalities.Method Congenital cardiovascular malformations of fetuses were diagnosed by prenatal ultrasonic cardiography from Jan 2011 to Sep 2013,and whose chromosomal karotype were tested by amniocentesis or cordocentesis.The association between chromosomal karyotypes and distribution of congenital cardiovascular malformations was analyzed.Result In 173 Fetuses with chromosomal abnormalities,20(11.56％) cases had congenital cardiovascular malformations,including seven 21-trisomies,eight 18-trisomies,three 13-trisomies and two 45,X.64％ (16/25) fetuses with congenital cardiovascular malformations accompanied with other malformations had chromosomal abnormalities.Only 1.87％ (52/4379) fetuses with normal karotype had congenital cardiovascular malformations.Conclusion Chromosomal abnormality is the most reason of complicate CHD.Chromosomal karotype test should be detected in fetus with complicate CHD.

Objective To explore the function of colonic mucosal barrier of patients with colonic slow transit constipation (STC).Methods From June 2008 to June 2012,a total of 136 patients with STC were enrolled.Among them,course of disease of 55 cases was between one and six years,of 43 cases was between six and 10 years,and of 38 cases was over 10 years.The colonic transit time of 66 cases was between three and five days,of 42 cases was between five and seven days,and of 28 cases was over seven days.Altogether,35 cases received subtotal resection of the colon.At same time,individuals who received partial resection of the colon because of single polypus were set as control group.Fasting blood and urine samples of all subjects were collected.The colonic specimens of STC patients who received surgery and control group were harvested.The urinary lactulose and mannitol ratio (L/M) was detected by high performance liquid chromatography (HPLC).The level of blood Dlactic acid (D-LAC) was tested by enzymatic spectrophotometric.The level of blood diamine oxidase (DAO) was determined by speetrophotometry.The level of endotoxin (ET) was detected by azo chromogenic substrate limulus test.The colonic epithelial cells membrane resistance (TER) and paracellular mannitol permeability (PMP) were measured with Ussing perfusion chamber.t-test was performed for comparison between groups.Results Urinary L/M of STC group and control group was 0.16±0.03 and 0.10±0.02,respectively.The level of blood D-LAC was (1.81±0.19) and (1.04±0.13) mmol/L.The level of blood DAO was (17.07±1.81) and (9.78±1.14) U/L.The level of blood ET was (64.20±6.85) and (51.30±5.90) EU/L.The TER of colonic epithelia cell was (61.23±7.76) and (75.87±9.65) Ω/cm2.The PMP of colonic epithelia cell was (3.17±0.35) ％ and (2.14 ±0.22)％.All the differences were statistically significant (t =3.185,3.378,3.863,3.201,3.125 and 3.543,all P＜0.05).Among patients with disease course between one and six years,six to 10 years and over 10 years,colonic transit time of STC between three and five days,five to seven days and over seven days,urinary L/M,blood D-LAC level and blood DAO level increased along with the disease course and colonic transit time and the differences were statistical significantly compared with control group (urinary L/M:t=1.993,2.311,2.356,2.204,2.347 and 3.673; blood D-LAC level:t=2.023,2.886,4.124,1.999,2.998 and 3.465; blood DAO level:t=1.994,2.995,4.423,2.203,3.673 and 5.211; all P＜0.05).Compared with control group,there were significant differences in blood ET level of course of STC between six and 10 years,over 10 years,colonic transit time of STC between five and seven days and over seven days (t=2.121,4.245,3.241 and 4.657,all P＜0.05).Conclusion The permeability of colonic mucosal barrier increased and which was more significant in longer colonic transit time and long course of disease.

BACKGROUND:Studies have reported that the superfamily of transforming growth factors exert a role in the mineralization of various stem cells, but the combination effects of transforming growth factorβ3 and heparin on proliferation and mineralization ability of stem cells from human exfoliated deciduous teeth remains to be studied. OBJECTIVE:To explore the effect of transforming growth factorβ3 on odontoblast-like differentiation of stem cells from human exfoliated deciduous teeth. METHODS:Human deciduous teeth were col ected using enzyme digestion. The 3rd cells were incubated with 25μg/L recombinant human transforming growth factorβ3, 10 U/mL heparin or their combination. The dentin sialophosphoprotein mRNA and dentinsialoprotein expressions were detected by Q-PCR and western blot assay. Alkaline phosphatase activity was determined using alkaline phosphatase kit. RESULTS AND CONCLUSION:Stem cells from human exfoliated deciduous teeth grew wel after induction. The activity of alkaline phosphatase in the combination group was significantly higher than that in the transforming growth factorβ3, heparin and control groups (P<0.01). After combination induction, the cells were strongly positive for alizarin red staining. Results fromα-PCR and western blot assay showed that the expressions of dentin sialophosphoprotein were both remarkably increased at mRNA and protein levels. In summary, stem cells from human exfoliated deciduous teeth can differentiate into odontoblast-like cells under the induction of transforming growth factorβ3 plus heparin.

BACKGROUND:The role of transforming growth factorβsuperfamily has been reported in bone mineralization of various types of stem cells, but the effects of transforming growth factorβ3 (TGF-β3) combined with heparin on proliferation and mineralization of dental pulp stem cells from human deciduous teeth remains to be studied. OBJECTIVE:To evaluate the effects of TGF-β3 on the proliferation and mineralization of dental pulp stem cells from human deciduous teeth. METHODS:The enzyme digestion method was utilized to separately culture dental pulp stem cells from human deciduous teeth. The cellcolony forming efficiency was determined. Flow cytometry was utilized to identify cellsurface marker CD146. Immunochemistry for Vimentin and STRO1 was performed to measure dental pulp stem cells from human deciduous teeth. The third passage dental pulp stem cells from human deciduous teeth cultured in vitro were intervened with heparin and TGF-β3 of 1, 5, 25μg/L mass concentration. The MTS method was applied to measure cellgrowth curves. Alizarin red staining was carried out. The changes in alkaline phosphatase activity were determined with alkaline phosphatase kit. RESULTS AND CONCLUSION:The cellcolony forming efficiency was high. cells were positive for CD146, and strongly positive for Vimentin and STRO1. Dental pulp stem cells from human deciduous teeth were identified. MTS assay indicated that there was no obvious effect on promoting proliferation of dental pulp stem cells from human deciduous teeth after stimulation of TGF-β3. Detection results of alkaline phosphatase activity demonstrated that the combination of TGF-β3 and heparin could strengthen the alkaline phosphatase activity of dental pulp stem cells from human deciduous teeth with increased concentration. Alkaline phosphatase activities were significantly higher in the TGF-β3+heparin group, TGF-β3 group and heparin group than in the control group (P<0.01). Alizarin red staining was positive in the TGF-β3+heparin group, and the staining was strongest in the 5μg/L TGF-β3+heparin group. Results indicated that TGF-β3 combined with heparin promoted mineralization of dental pulp stem cells from human deciduous teeth.

Objective To investigate the significance of orthotopic liver transplantation in the management of primary hepatocellular carcinoma (PHC) ; To evaluate the survival and prognostic factors of liver transplantation for PHC.Method The clinical data from 194 consecutive PHC patients who underwent OLT were retrospectively analyzed.Result The 1-,3-and 5-year cumulative survival rate of the 194 patients was 92.78％,78.07％ and 76.80％ respectively.There were 37 cases of recurrence or metastasis of the tumor with the incidence rate being 19.1％ (37/194).The tumor TNM stage (P=0.026 667),tumor diameter (P =0.0045),tumor number (P =0.0318),vascular invasion (P =0.006 059),and AFP level before transplantation (P =0.003 426) were significantly related with tumor recurrence.Multivariate analysis revealed that tumor diameter (P =0.0081,OR =0.789,95％ CI:2.1233-6.124),vascular invasion (P＜0.001,OR =5.928,95 ％ CI:1.173-86.789),and AFP level before transplantation (P =0.019,OR =2.601,95％ CI:2.196-5.658) were independent risk factors for tumor recurrence.There was statistically significant difference in survival rate and tumor-free survival rate between Milan criteria group and beyond-UCSF criteria group (P＜0.01),and between UCSF criteria group and beyond-UCSF criteria group (P＜0.01).There was no significant difference in survival rate and tumor-free survival rate between Milan criteria group and UCSF criteria group (P ＞0.05).Conclusion Liver transplantation is an effective treatment for PHC.The tumor diameter ＞8 cm,vascular invasion and AFP level significantly affect the survival of PHC patients.

Objective To determine the difference of macrophage RAW264.7 apoptosis induced by Brucella melitensis virulent strain 16M and attenuated strain M5-90 and elucidate the regulatory role of caspases 3,8 and 9.Methods The best multiplicity of infection (MOI) was determined through kinetic analysis of Brucella melitensis strain 16M and M5-90 induced mouse macrophages apoptosis(bacterium ∶ cell =100 ∶ 1,50 ∶ 1,10 ∶1).The infection model was established using the best MOI =50 ∶ 1.The numbers of in vivo bacteria by colony formation units were calculated after macrophages were infected for different times,including 2,4,8,12,24 and 48 h,and the infected cells were collected.The ratios of apoptosis were detected and the regulation of caspases 3,8 and 9 in apoptosis pathway was elucidated by flow cytometry.Results The numbers of 16M in vivo bacteria were 105.4,104.8,105.8,106.5,108.0 and 109.0,respectively and of M5-90 were 106.1,106.2,106.4,106.3,106.1 and 105.0,respectively.The number of in vivo bacteria of 16M was significantly increased than that of M5-90 after infected for 24 h to 48 h.The ratios of apoptosis induced by 16M after infected for 2,4,8,12,24 and 48 h was (2.67 ± 0.09)％,(13.13 ± 0.30)％,(6.56 ± 0.42)％,(6.49 ± 0.28)％,(16.07 ± 0.86)％ and (24.23 ± 1.67)％,respectively,and by M5-90 was (3.62 ± 0.02)％,(32.01 ± 2.59)％,(17.58 ± 0.44)％,(16.09 ± 0.10)％,(62.53 ± 2.70)％ and (85.53 ± 0.15)％,respectively,and by control group was [(1.90 ± 0.20)％,(1.92 ±0.16)％,(1.99 ± 0.03)％,(2.48 ± 0.11)％,(3.56 ± 0.07)％,(5.26 ± 0.33)％].The differences were statistically between groups in same time.The Brucella melitensis vaccine strain M5-90 was more powerful than virulent strain 16M in respect of inducing macrophage apoptosis after infected for 24 to 48 h.Twenty-four hours after infection,the expression of caspases 3,8 and 9 was (1.47 ± 0.05)％,(1.52 ± 0.02)％ and (2.47 ± 0.12)％,respectively,in control group and the expression was (9.70 ± 0.46)％,(6.08 ± 0.56)％ and (35.08 ± 1.64)％,respectively,after infected for 24 h induced by M5-90.The expression of caspases 3,8 and 9 was significantly higher than that control group (P ＜ 0.01).Twenty-four hours after given caspases 3,8 and 9 inhibitor,apoptosis rate in control group was (66.72 ± 1.28)％,in M5-90 group was (22.58 ± 0.55)％,(53.15 ± 1.85)％ and (29.18 ± 0.23)％,respectively,and compared with control group,apoptosis rate of caspases 3,8 and 9 was significantly lower(P ＜ 0.01).Conclusions Apoptosis of macrophage can be induced by Brucella melitensis virulent vaccine strain 16M and attenuated strain M5-90.M5-90 is stronger than that of strain 16M.Caspases 3,8 and 9 can regulate macrophage apoptosis after M5-90 infection.