In this study ,the objective is to establish a nested‐PCR assay for the detection of H .bilis with high sensitivity and specificity .The nested primers were designed based on sequences of 16S rRNA gene of seventeen subtypes of H .bilis .Af‐ter optimizing reaction condition ,the sensitivity and specificity of the assay were examined via the detection of feces simulated samples ,mice infection model samples and clinic patients’ samples .The detection sensitivity of H .bilis strain for feces simu‐lated samples was 10 CFU/100 μL .H .bilis was successfully detected in the liver ,caecum and feces of experimentally infected mice .Moreover ,H .bilis was successfully detected in the bile ,cholecyst mucous membrane and feces samples from two of ten patients with cholelithiasis .Due to the PCR assay’s high sensitivity and specificity ,the method may be used to detect the infec‐tion of H .bilis .

Objective To discuss the method, safety and clinical value of biliary stenting combined with 125I seed implantation intracavitary irradiation in treating malignant obstructive jaundice. Methods A total of 36 patients with malignant obstructive jaundice were enrolled in this study. PTCD was carried out in all patients, which was followed by biliary stenting combined with 125I seed implantation intracavitary irradiation treatment. The results were analyzed. Results During the interventional management, displacement of the stent and 125I seeds were observed in two cases, and the displaced stent and 125I seeds were replaced to the right position with the help of biliary biopsy forceps. The technical success rate was 100%, and the remission rate of the jaundice was 100%. All the patients were followed up for 1-23 months. No radioactive particles leaking or complications such as radiation enteritis occurred. No in-stent obstruction due to tumor recurrence was observed although slight dilatation of intrahepatic bile duct was detected in 25%of patients, which was resulted from intimal hyperplasia at the stent mesh and/or biliary stone formation. The median survival time was 10.9 months. Conclusion For the treatment of malignant obstructive jaundice, biliary stenting combined with 125I seed implantation intracavitary irradiation is safe, reliable and effective. This technique can prolong stent patency time as well as the patient’s survival time.

Objective To study the iodine nutritional status of population living in Tibetan pastoral areas,in order to provide a scientific basis for prevention and control of iodine deficiency disorders.Methods Drinking water samples were collected to test iodine content in agricultural town(Kajiaman) and pastoral area(Zuogaiduoma town) of Hezuo in Gannan Tibetan autonomous prefecture.Thirty of child-bearing age,pregnant and breastfeeding women were selected,respectively,and 90 male adults aged 20-50 from these families(1 from each family) and 90 children aged 8-10 (30 people in each age group) from local schools were randomly sampled at the same time,and urinary iodine (UI) was measured randomly.Edible salt and main food samples were collected to test iodine content from the 10 families of the three types of women,respectively,and they were asked to recall its family intake of food species in the past 24 h excluding spices.The water iodine was determined using arseniccerium redox method (GB/T 5750.1-2006) ; UI with ammonium persulfate digestion-arsenic cerium catalytic spectrophotometric method (WS/T 107-2006) ; salt iodine used direct determination method(GB/T 13025.7-1999); and food iodine with alkali the gray arsenic cerium contact colorimetry.All these work were done in May,2011.Results The average of water iodine was (1.63 ± 0.14)μg/L in agricultural areas and (2.08 ±1.90)μg/Lin pastoral areas of the 10 water samples tested,respectively.The median urinary iodine(MUI) among women of pregnant,lactating and child-bearing age,male adults and children was 141.99,126.65,253.33,258.07,191.0μg/L,respectively,in agricultural areas and 137.26,97.36,126.16,159.48,285.07 μg/L,respectively,in pastoral areas.The difference of MUI in lactating,male adults and children between pastoral and agricultural areas was statistically significant.The proportion of UI ＜ 50 μg/L was less than 20％,and ＜ 100 μg/L was less than 50％ among all population except lactating woman and pregnant women in pastoral areas.The iodized salt coverage rate was 100％(30/30) in agricultural areas and 90％(27/30) in pastoral areas,and the salt iodine was (32.1 ± 7.8)mg/kg in agricultural areas and (32.3 ± 6.0)mg/kg in pastoral areas,respectively.The food structure in agricultural areas was mainly potato,naked oat fruit,cabbage and so on,the average dietary iodine content was 285.7 μg/kg,and in pastoral areas was mainly chow mein,wheat flour,ghee,yogurt,barley and so on,the average dietary iodine content was 51.1 μg/kg.Conclusions There is no iodine deficiency in general in the population in Tibetan areas with low water iodine.However,iodine nutrition of pregnant women can not be guaranteed.It is recommended that classified guidance measures be taken to ensure the sustainable elimination of iodine deficiency disorders in the Tibetan minority areas.

Objective To further investigated the effect of minocycline on the inhibition of microglial activation and subsequent protection of nigral DA neuron.Methods 20 rats injected with LPS in the substantia nigra (SN) were randomly divided into two groups (LPS group and LPS+Minocycline group).The behavior was observed on the 7~(th) d and 14~(th) d.The immunohistoehemistry,in situ hybridization and Western-blot were used to detect the levels of positive neuron,mRNA,protein of TH and OX-42. Results The slightly rotational behavior was observed in LPS+Minoeyeline group.The majority of mieroglias were activated in the two groups.Some microglia in the SNpc remained ramified in LPS+ Minocycline group.The numbers of hypertophie microglia in LPS+Minoeyeline group were less than that in LPS group.Western-blot showed that the protein of OX-42 in two LPS groups was higher than in normal group(P

BACKGROUND:As a noticeable tissue engineering material of polyhy droxyalka noates family, poly (3-hydroxybutyrate-co-4-hydroxybutyrate)(P3HB4HB) exhibitsgood biocompatibility, adhesion and mechanicalproperties, presenting aextensive application future in tissue-engineered research.
OBJECTIVE:To investigate the biocompat ibilityin vitroand ectopic osteogenic differentiationin vivoof P3HB4HB and human bone marrow mesenchymal stem cels.
METHODS:Passage 5human bone marrow mesenchymal stem cels transplanted ontothe three-dimensional P3HB4HB scaffoldwereincubated with osteogenic induction medium (test group)or with no osteogenic induction(control group), respectively. After 5-day incubation, thecelgrowth was assessed by acridine orange staining and scanning electron microscopy; after14-day incubation, both kinds of cel-scaffold composites were subcutaneously implanted into the nude mice. At 16 weeks after implantation, the cel-scaffold composites were removed to observeectopic osteogenic differentiationin vivousing hematoxylin-eosin staining, von Kossa staining and colagen type I immunohistochemical staining.
RESULTS AND CONCLUSION:Acridine orange staining showed that cels adhered wel on the surface of the scaffold;under thescanning electron microscope, induced celsgrew wel on the P3HB4HB scaffold and produced abundant extracelular matrixes. In addition, at 16 weeks after implantation, there were osteoidtissues in the test group, positive for von Kossa staining as wel as colagen type I immunohistochemical staining;furthermore, hematoxylin-eosin staining showednumerous osteoblasts and bone lacunas. In contrast, no bone tissues appeared in the control group. To conclude, P3HB4HB is a suitable material for bone tissue engineering.

Background Corneal neovascularization (CNV) is an important cause of visual impairment and graft rejection after allograft corneal transplantation in inflammatory corneal diseases. The mechanisms and therapy relating to CNV are intensely investigated at all times. Objective This study was to evaluate the effect of eicosapentaenoic acid (EPA) on CNV induced by alkali cauterization and its mechanism. Methods The animal models of corneal neovasculation were induced in the right eyes in 72 Sprayue-Dawley rats by putting a piece of 3 mmfilter paper with 1 mol/L NaOH at the center of the cornea for 30 seconds. The rats were then divided randomly into the 0.02 mg EPA treatment group (24 rats) ,0.03 mg EPA treatment group (24 rats) ,model group (24 rats) and normal group (6 rats). EPA of 0.04 ml with doses of 0.02 mg or 0. 03 mg or saline solution of 0. 04 ml was injected subconjunctivally in model rats and immediately after cauterization. The presence of CNV and corneal edema were observed daily by slit lamp biomicroscope. 1,4,7 and 14 days after operation, corneal histopathological examination was performed by hematoxylin and eosin staining. The vascular endothelial cells were stained with CD34 by immunohistochemistry,and the expression of IL-1α,IL-6 mRNA and the nuclear factor-κBp65 ( NF-κBp65 ) proteins was measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. The use of animals complied with the Regulations for the Administration of Affair Concerning Experimental Animals by Hebei Province( version 1998 ). Results Under the slit lamp, CNV grew slowly from days 2-4 with obvious corneal edema and defect of epithelium. Larger CNV area and less edema were seen from days 7-10. Maximal vessel growth was observed 14 days after injury with thinner vessels in the model group. Histological examination showed that part of the corneal epithelium was damaged;serious corneal edema, more inflammatory cells and a lot of CNV in the stroma were presented in the model group. However, repairing of the corneal epithelium without CNV ,light corneal edema and less inflammatory cells were found in both the 0. 02 mg EPA and 0. 03 mg EPA treatment groups 7 days after alkali cauterization. The relative area of CNV in the 0. 02 mg EPA treatment group was ( 15.80±6.43 )% and ( 11.06±2. 14)% ,and that in the 0. 03 mg EPA treatment group was (16. 10±7.41 )% and (11.06±2. 51 )%, showing significant reduction in comparison with the model group [ (84. 74±7.77)% and (89.63±7.50) % ] 7 days and 14 days after operation ( P＜0. 05 ). Stronger expression of CD34 in the vascular endothelial cells of the cornea stroma was observed in the model group and an absence of CD34 was observed in the EPA-treated groups on the 7th day. RT-PCR revealed that the expression of IL-1α mRNA and IL-6 mRNA was lower in the EPA treatment groups than the model group ( P＜0. 05 ), and Western blot analysis showed that the expression of NF-κB/p65 in the corneas in the EPA treatment groups was significantly lower than that in the model group on the 4th day after operation (P＜0.05).Conclusion Topical application of EPA suppresses CNV induced by alkali burn possibly by inhibiting the expression of NF-κB,IL-1α and IL-6.

To establish a method for the determination of ketamine and MDA and their main metabolites in urine by solid phase microextraction-liquid chromatography-mass spectrometry. In a urine sample supplemented with quantitative ketamine, norketamine, MDMA and MDA control. The solution was adjusted pH 11, added solid Na2CO3, heated and stirred at 60 °C. Then, the sample was extracted by SPME with 60 μm polydimethylsiloxane-vinylbenzene copolymer(PDMS/DVB ), a middle-polar coated fiber for 15 minutes and then analyzed by HPLC-MS. The result showed good linearity in the range of 0. 03-1. 0 μg/mL, r≥0. 999 2, and LOD was 0. 01 μg/mL, the value of the average recovery rate was varying from 97. 19%-105. 44%, and RSD was within 10%. The method is simple, safe and accurate, and can be used to determine ketamine, MDMA and their main metabolites in urine.

Objective: To study the mechanism of inhibitory ef fect of ginsenoside Rg3 on tumor growth. Methods: The chick chor ioallantoic membrane(CAM) test and Lewis lung carcinoma model were used to inves tigate the inhibitory effect of Rg3 on tumor angiogenesis. Results: Rg3(0.1 or 0.5 mmol/L) obviously inhibited angiogenesis in the CAM. Treatmen t with Rg3 in vivo obviously inhibited Lewis lung carcinoma growth with the inhibition ratio from 23% to about 47%. We also observed that the angiogenesis in implanted Lewis lung carcinoma tissue decreased obviously after treated wit h Rg3 (5, 10, 20 mg/kg). Conclusion: Rg3 can obviously inhibit t he growth of Lewis lung cancer, the inhibitory effect partially due to the effec t of Rg3 inhibiting neovascularization induced by malignant tumor.

Abdominal Pain ; etiology ; Acute Disease ; Adult ; Atropine ; therapeutic use ; Chemical and Drug Induced Liver Injury ; etiology ; therapy ; Diarrhea ; etiology ; Fatal Outcome ; Female ; Humans ; Male ; Middle Aged ; Multiple Organ Failure ; etiology ; Muscarinic Antagonists ; therapeutic use ; Mushroom Poisoning ; complications ; therapy

To evaluate the efficacy of an ultrasound-based quantitative method to diagnose liver fibrosis using a rat model. Ultrasonography was performed on the livers of 90 Sprague-Dawley rats with or without thioacetamide-induced fibrosis. The liver capsule thickness and 13 texture parameters of gray level co-occurrence matrix were extracted from the standard sonograms. After sacrifice, severity of liver fibrosis (S0-S4 classification) was diagnosed by histopathology. Analysis of variance and correlation statistical tests were used to analyze the differences between groups and determine the relationships between each of the 14 quantitative ultrasound index points and the histological results, respectively. Discriminant analysis models were developed for quantitative diagnosis of liver fibrosis, and the leave-one-case-out method was used to verify the efficiency of models. All 14 indices were significantly correlated with the histological stages of fibrosis (P less than 0.05). The accuracy of the discriminant model for S0, S1, S2, S3 and S4 was 83.3%, 84.2%, 70.0%, 50.0% and 88.2%, respectively. In addition, 73.3% of cross-validated rats were accurately classified. Grouping S0 as no fibrosis, S1 as mild fibrosis, S2 with S3 as moderate to severe fibrosis and S4 as early cirrhosis increased the accuracy of the discriminant model for these four groups (respectively, 91.7%, 84.2%, 69.0% and 88.2%) and allowed for 78.9% of cross-validated rats to be correctly identified. Ultrasonography combined with texture analysis was a novel and accurate method to diagnose liver fibrosis in a rat model; further studies may provide insights into its applicability for quantitating liver fibrosis in other animal models or in clinic.
Animals ; Liver ; diagnostic imaging ; pathology ; Liver Cirrhosis, Experimental ; diagnostic imaging ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Ultrasonography