This study aimed to investigate the anti-apoptotic effects of Tangnaikang (TNK) on islet β cells in Zucker diabetic fatty (ZDF) rats.Six male fa/+ ZDF rats were took as the control group,while other thirty male fa/fa ZDF rats were divided into five groups at random:the model group,the metformin group,the high-,mediumand low-dose TNK groups,depending on their body weight and random blood glucose.Prior to the administration,fasting blood glucose and fasting insulin were measured by drawing blood with inner canthusl.Materials were prepared when administered for six weeks.Fasting blood glucose and fasting insulin were detected again.When the sections of the rat pancreatic tissue were embedded,the morphological changes of the islet were observed via HE staining,and the apoptosis of islet β cell were observed using TUNEL.Positive expression of Caspase-3,the transduction enzyme of cell death signal,was tested by immunohistochemical method.It was found that the fasting blood glucose of the (fa/fa) ZDF rats in the high-,medium-and low-dose TNK group was significantly improved after administration (P ＜ 0.01).The serum insulin of rats in the high-,medium-and low-dose TNK group arised compared with the model group,while the high-and low-dose TNK group showed differences in a statistical sense.Compared with the model group,the HOMA-IR of all the treatment group decreased,while significant difference was presented between the high-dose TNK group and the metformin group.HE staining showed that the morphology of the islet β cell of the rats in all the treatment group was improved.The results of TUNEL showed significantly apoptotic changes on islet β cell of the fa/fa ZDF rats.Compared with the model group,the positive expression of TUNEL in the metformin group and the high-dose TNK group were significantly reduced (P ＜ 0.05).The result of immunohistochemistry method showed that the protein levels of Caspase-3 in the metformin group and the high-dose TNK group decreased (P ＜ 0.05).In conclusion,it was demonstrated that TNK effectively reduced the apoptosis of islet β cells in fa/fa ZDF rats,which presented a protective effect.

The contents of coumarins in the sulfur fumigated Angelicae Dahuricae Radix (Baizhi, ADR) were reduced significantly. To achieve the quality control of ADR, the qualitative identification of sulfur fumigated ADR and quantitative model of imperatorin content should be established. The near-infrared (NIR) spectrograms of non-sulfur and sulfur fumigated ADR were collected by NIR diffuse reflectance spectroscopy technology and pretreated by the method of first derivative derivation and vector normalization. The Ward's Algorithm method was used for the cluster analysis. The non-sulfur and sulfur fumigated ADR can be quickly identified in the range of 8,806. 0-3 811.0 cm(-1) based on the cluster analysis. The NIR quantitative model of imperatorin was established by the contents of imperatorin determined by HPLC in combination with partial least squares regression analysis. According to the calibration model established in this study, correlation coefficients (R2), the root-mean-square error of cross-validation (RMSECV), and the root-mean-square error of prediction (RMSEP) for imperatorin were 0.982 8, 0.006 8, 0.011 8, respectively. The quantitative model of imperatorin can be applied to determine the content of imperatorin in ADR accurately.
Angelica ; chemistry ; classification ; China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Quality Control ; Spectroscopy, Near-Infrared ; methods ; Sulfur ; chemistry

Objective To study whether sulphur fumigation can cause changes to the content of imperatorin in Angelicae Dahuricae Radix (Baizhi).Methods The fresh samples of Angelicae Dahuricae Radix were collected from some cultivation bases. Half of each batch root was fumigated with sulfur according to the conventional method, and another half was cut into slices and dried naturally (i.e. without sulfur fumigation). The content of imperatorin was determined by HPLC on C18 column (4.6 mm×250 mm, 5μm) with a gradient mobile phase of acetonitrile-water solution system at the follow rate of 1.0 mL/min, 35℃ of the column temperature, and the max plot in 210-800 nm of the detective wavelength.ResultsTotally 29 batches of fresh roots ofAngelica dahurica were collected. The average contents of imperatorin were 0.202% in the Angelicae Dahuricae Radix with sulfur fumigation and 0.120% in the Angelicae Dahuricae Radix without sulfur fumigation. Compared with the corresponding samples without sulfur fumigation, the content of imperatorin in every Angelicae Dahuricae Radix with sulfur fumigation decreased by 6.77%-77.56% with an average decrease of 39.86%.Conclusion The content of imperatorin decreased significantly in every batch of Angelicae Dahuricae Radix with sulphur fumigation. It shows that the sulphur fumigation method was not suitable for producing and processing Angelicae Dahuricae Radix from fresh roots ofA. dahurica.

Objective To investigate an optimum extracting technology for total glycosides from dried barks of Ilex rotunda Thunb. Methods The yields of pedunculoside and syringin by high performance liquid chromatography (HPLC) determination were taken as the indexes. Some parameters of the extraction technology were evaluated with an L9(3)4 orthogonal design. The optimum extraction parameters were used to extract total glycosides in laboratory. The feasibility was checked by determining the chemical constituents by means of HPLC method. Results The optimum extracting conditions were 12 times volumes of 50% ethanol as the solvent and refluxing 90 min each time for 3 times. An extract yield of total glycosides was 19.5% from barks of Ilex rotunda. Pedunculoside (292 mg/g), syringin (59.5 mg/g), sinapaldehyde glucoside, syringaresinol 4'-O-β-D-glucopyranoside, and syringaresinol 4',4'-di-O-β-D-glucopyranoside were detected in the extract. Conclusion Some glycosides were found in the extract with the optimum extraction technology in laboratory. The extraction technology is practicable and valid.

Objective To construct the serine protease gene(sprE)mutant and to study the pathogenicity of sprE gene of Enterococcus faecalis.Methods Recombinant suicide vector pCQ001 of Enterococcus faecalis with pTX4577,was constructed.Then,created isogenic sprE-deficient mu- tant(*sprE)by allelic replacement was constructed.Moreover,the growth ability and the virulence of the mutant were compared with those of the wide type in vitro and in vivo respectively.A mouse peritonitis model and a rabbit endocarditis model were utilized in the study.Results The *sprE was selected by kanamycin and identified by polymerase chain reaction(PCR),pulsed field gel electropho- resis(PFGE)and Southern blot.The evidences showed that the sprE gene had a major role in helping bacteria to resist the elevated temperature and oxidative stress.The virulence of mutant decreased af- ter sprE gene was knocked out.Conclusions The *sprE of Enterococcus faecalis is constructed suc- cessfully,sprE gene is important in the pathogenesis of Enterococcus faecalis,which probably is a major virulence factor of Enterococcus faecali.

Objective To investigate the effect of diabetes on clinical efficacy of transcatheter arterial chemoembolization (TACE) in the treatment of non-viral hepatitis hepatocellular carcinoma (HCC). Methods Retrospectively analyzed the clinical data of 367 non-hepatitis virus HCC patients treated by TACE, included 153 diabetes mellitus cases (test group) and blood glucose of 214 patients was normal (control group). To assess the treatment effect after 1 month of TACE based on response evaluation criteria in solid tumors, include complete response (CR), partial response (PR), stable disease (SD), progressive disease (PD), and calculate the disease control rate. Through 6 to 75 months follow-up to observed long-term efficacy, record the time to progression (TTP) and overall survival (OS) time. Survival rate were analyzed using Kaplan-Meier method and Log-rank analysis by SPSS 16.0. The single-factor analysis was used to analyze variables which variables that differed were analyzed by Cox regression. Results The disease control rate of test group was 69.9%(107/153) and control group was 74.3%(159/214), the difference was no statistically significant (P=0.125). The median time to progression (mTTP) and median overall survival (mOS) of test group were 10.0 and 15.0 months;and the mTTP and mOS of control group were 14.0 and 19.0 months, the difference were statistically significant (P=0.023 and P= 0.026). Tumor diameter ≥4.5 cm, numbers of tumor ≥3, invasion of blood vessels, α-fetoprotein≥200 μg/L, Eastern Cooperative Oncology Group score and diabetes were risk factors for OS of HCC patients. Conclusion Diabetes is unfavorable factors for overall survival of non-hepatitis HCC tread by TACE.

Objective To investigate the effect of acupuncture combined with minimally invasive hematoma aspiration on a rabbit model of brain injury in the acute stage of intracerebral hemorrhage (ICH). Methods A total of 65 healthy New Zealand white rabbits were randomly divided into a sham operation (SO) group, an ICH group,a hematoma aspiration (HA) group, and an acupuncture and hematoma aspiration (AHA) group. Models of ICH were established in the latter three groups. The HA group was treated with minimally invasive hematoma aspiration and the AHA group was treated with both acupuncture and minimally invasive hematoma aspiration. At 6 hours and 1, 3 and 7 days after the ICH models were established, brain water content (BWC) was measured, and the expression of matrix metalloproteinase-9 ( MMP-9 ) in the rabbits' brains was detected by immunohistochemistry. MMP-9activity was detected by gelatin zymography. Results The BWCs of the ICH group, HA group and AHA group rabbits were significantly higher than those of the SO group. The BWCs of the HA group and AHA group animals descended significantly more than those in the ICH group as time went on, especially on the 3rd day in the AHA group. Immunohistochemistry and gelatin zymography showed that the expression and activity of MMP-9 in these test groups decreased with time, especially on the 3rd day in the AHA group. Conclusions Acupuncture combined with hematoma aspiration can reduce injury in the acute stage of ICH, and inhibition of the expression of MMP-9 may be the mechanism.

Objective To study the distribution patterns of bacterial flora in sigmoid colon tissues and stools in normal population.Method Bacterial flora were identified and analyzed by using 16sDNA sequencing technology in fresh stool samples (n =13) and colon mucosa samples (n =10).Results The diversity and abundance of bacterial flora were significantly larger in the stool samples than in the sigmoid colon samples (P ＜ 0.001,P ＜ 0.001,P =0.042,P =0.006).The consititution of phylum flora between the two groups were same,including flrmicutes,bacteroides,proteobacteria,and actinomycetes.However,the proportions of firmicutes and bacteroides in stool samples were significantly higher than in the sigmoid colon samples,whereas the proportion of proteobacteria was significantly lower (P ＜ 0.001,P =0.025,P ＜ 0.001).At the genus level,faecalibacterium and bacteroides were the dominant flora in feces,whereas pseudomonas,lactococcus,acinetobacter,and flavobacterium were the most common flora in sigmoid colon mucosa.The amounts of bifidobacterium and lactobacillus were low in both two groups.Conclusion The distribution of bacterial flora remarkably differ in stools and sigmoid colon mucosa.

Objective To investigate the nuclear factor-erythroid 2-related factor (Nrf-2),and heme oxidase 1 (HO-1) expression in acute lung injury induced by paraquat poisoning in rats and explore the mechanism of lipoic acid acting on protection of lung from paraquat poisoning.Methods Seventy-two adult healthy male Sprague Dawley (SD) rats were randomly divided into three groups with different treatments designated as:control group (control group,n =12),paraquat group (PQ group,n =30) and paraquat + lipoic acid group (LA group,n =30).PQ group and LA group were randomly divided into five subgroups (n =6 in each) according to 6 h,12 h,24 h,48 h and 72 h after modeling and treatment.The rats in PQ group and PQ + LA group were treated with intra-peritoneal injection (ip) of PQ (25 mg/kg),while the rats in control group were treated with the equal volume of saline instead.Half an hours after intra-peritoneal injection of PQ,lipoic acid (100 mg/kg) was injected into caudal vein of rats once a day until they were sacrificed.The body weight was measured everyday.The rats of each group were sacrificed at the given intervals,and lung tissues were harvested to measure lung coefficient of rats.The same part of left lung of rats in each group was taken for HE staining and immunohistochemistry in order to detect the expressions of Nrf2 and HO-1.The right lung of rats in each group was taken for the detection of GSH-Px and SOD activity.All data were analyzed by using the One-way analysis of variance (ANOVA) and SNK-q test.Results The body weight reduction in LA group (191.02 ± 0.82) g,(183.37 ± 7.74) g was significantly less than that in PQ group (183.85 ± 2.07) g,(173.13 ± 4.34) g at 48 h and 72 h after PQ poisoning,respectively (P ＜ 0.01,P ＜ 0.05).The lung coefficient in LA group (6.83 ± 0.48) mg/g,(7.61 ±0.28) mg/g,(8.29 ±0.36) mg/g was less compared with PQ group (7.39 ±0.53) mg/g,(8.48±0.23) mg/g,(9.06±0.10) mg/g at 24 h,48 h,and 72 h,respectively (P＜0.01,P＜ 0.05).The immunohistochemical expressions of Nrf-2 in LA group (3.99 ±0.50),(3.51 ±0.12) were higher than those in PQ group (1.33 ±0.22),(1.62 ±0.41) at 48 h and 72 h.The immunohistochemical expression of HO-1 in LA group (1.76 ±0.17) was higher than that in PQ group (1.31 ±0.15) at 72 h.The levels of GSH-Px activity in LA group were significantly higher in comparison with PQ group at 24h,48h,and 72h (P ＜0.01,P ＜0.05).The levels of SOD activity in the LA group were significantly higher in comparison with PQ group at 6 h,12 h,24 h,48 h,and 72 h after PQ administration (P ＜ 0.01).Conclusions Nrf2-ARE (antioxidant response element) signaling pathway is involved in the pathogenesis of acute lung injury induced by paraquat poisoning,and lipoic acid may protect acute lung injury in rats induced by paraquat poisoning through Nrf2-ARE signaling pathway.

Objective:To investigate the characteristics of short-term prognostic factors in very elderly patients with acute coronary syndrome (ACS).Methods:A total of 2 912 ACS patients admitted to Shengjing Hospital of China Medical University from January 1, 2010 to October 31, 2014 and treated with percutaneous coronary intervention (PCI) were enrolled and divided into two groups according to age: very elderly group (≥75 years, 480 cases) and control group (< 75 years, 2 432 cases). The clinical data and coronary artery lesions of the included patients were detected. Major cardiovascular adverse events (MACE) occurred within 30 d after discharge were followed up and recorded. The short-term prognostic factors in very elderly patients with ACS were analyzed by Logistic regression.Results:Compared with control group, the percentage of hypertension, global registry of acute coronary events (GRACE) score, high density lipoprotein cholesterol, N-terminal pro-brain natriuretic peptide, the left main lesion ratio and Gensini score in very elderly group were higher, while the percentage of men, number of smoking, hyperlipidemia proportion, red blood cell count, white blood cell count, blood platelet count, albumin, and long term oral administration of aspirin, clopidogrel, statins, angiotensin receptor inhibitor after discharge were lower, and the differences were statistically significant ( P<0.01 or<0.05). During the follow-up period, the all-cause mortality in very elderly group was higher than that in control group: 2.5%(12/480) vs. 0.9% (21/2 432), and the difference was statistically significant ( P<0.01). Multivariate analysis showed that oral angiotensin converting enzyme inhibitor was a protective factor for elderly ACS patients after discharge ( OR = 0.046, 95% CI 0.006 to 0.383, P = 0.004). The receiver operating characteristic curve analysis showed that Gensini score ≥ 87.75 scores was a threshold value for all-cause mortality. The all-cause mortality ratio in high Gensini score (≥ 87.75 scores) group was higher than that in low Gensini score (<87.75 scores) group: 6.6% (9/137) vs. 0.9% (3/343), and the difference was statistically significant ( P<0.01). Conclusions:Very elderly patients with ACS have their own characteristics from both clinical history and prognostic factors. Patients with Gensini score of ≥ 87.75 scores should be closely observed, and drug treatment during hospitalization should be intensified if necessary. Follow-up should be strengthened for such patients, and oral drug treatment should be continued after discharge.