Objective To study the role of kallikrein 8 (KLK 8) in the pathogenesis of cerebral white matter injury induced by intrauterine infection.Method The pregnant Sprague Dawley rats were randomly assigned into two groups:the observation group and the control group.The rats in the observation group received intraperitoneal injection of lipopolysaccharide (500 μg/kg) by at day 18 and 19 of pregnancy,while the control group received the same dose of saline.The morphology of white matter of the newborn rats were observed at 1 d,3 d,7 d and 14 d after birth.The expression of KLK 8 in the hippocampus was examined using Western blot and reverse transcription-polymerase chain reaction (RTPCR);the concentration of KLK 8 in the serum was measured using enzyme linked immunosorbent assay (ELISA) method at the same time.Result In the observation group,the brain tissue was loose and edematous,the cerebral white matter was weakly stained,and the number of cells reduced.The expressions of KLK 8 in hippocampus in the observation group were higher than the control group [1 d:(0.24 ±0.01) vs.(0.23±0.01),3 d:(0.72±0.02) vs.(0.55±0.04),7 d:(1.08±0.04) vs.(0.84±0.04)],the differences were statistically significant (P ＜ 0.05).The expressions of KLK 8 mRNA in hippocampus of the observation group [1 d:(0.013 ±0.003),3 d:(0.032 ±0.002),7 d:(0.060 ±0.005)] were higher than the control group [1 d:(0.008 ±0.002),3 d:(0.016 ±0.002),7 d:(0.026 ±0.002)],the differences were also statistically significant (P ＜ 0.05).The serum KLK 8 concentration at 1 d,3 d,and 7 d were (5.13 ±0.24) μg/L,(6.46 ±0.24) μg/L,and (7.77 ±0.30) Iμg/L in the observation group,higher than the control group (4.73 ±0.25) μg/L,(5.65 ±0.29) μg/L,and (6.66 ±0.46) μg/L),the differences were also statistically significant (P ＜ 0.05).Conclusion KLK 8 may be involved in the pathogenesis of white matter injury induced by intrauterine infection.

With the development of imaging technique and molecular biological technique,researchers all over the world have made a great many of studies on the mechanism and treatment of lymphatic metastasis.The discovery of lymphatic endothelial cell-specific markers make it clear to distinguish lymphatic endotheliocyte.The application of new techniques such as immunohistochemical method and reverse transcription polymerase chain reaction (RT-PCR) can study the lymphatic metastasis from the genetic and molecular level.The application of molecular imaging and nanoparticles could provide new detection methods and therapies.The establishment of animal models offers a solid foundation for experimental study.

Objective To analyze the reasons causing false positive results of white blood cells (WBC) detected by AVE‐763B automatic urine analyzer .Methods WBC positive samples ,detected by AVE‐763B automatic urine analyzer ,were re‐inspected by microscope .Results In the 740 positive samples ,224 cases were false positive ,the false‐positive rate was 30 .2% ,among which 145 cases could be correctly classified by using image analysis ,and the false‐positive rate reduced to 10 .7% .Conclusion Many fac‐tors could cause false positive results .To improve the accuracy of AVE‐763B urine analyzer ,re‐inspect should be performed .

Objective To investigate the content of intestinal fatty acid binding protein (IFABP) and its clinical significance in patients with severe sepsis.Methods A prospective observational study was conducted.Fifty patients with severe sepsis admitted to intensive care unit (ICU) of the First Affiliated Hospital of China Medical University from July to December 2012 were enrolled,and 20 healthy patients served as control group.The concentrations of serum IFABP,interleukin-6 (IL-6),and tumor necrosis factor-α (TNF-α) were determined with enzyme-linked immunosorbent assay (ELISA) on days 0,1 and 3 after ICU admission.Acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) and sequential organ failure assessment (SOFA) score,28-day prognosis,acute gastrointestinal injury (AGI) grade were recorded at the same time.Furthermore,the contents of IFABP were compared between control group and the severe sepsis group,abdominal infection group and non-abdominal infection group,the survival group and the death group,as well as among different AGI-grade groups.Correlation analysis of IFABP and inflammatory factors,IFABP and two scores,and IFABP and time of stay in ICU and mechanical ventilation were studied.Multivariate logistic regression and analysis of 28-day outcome of the patients were also studied.Results IFABP levels were increased in severe sepsis patients on days 0,1 and 3 compared with those of healthy control group (mg/L:731.90 ±53.91,592.07 ±41.94,511.85 ±47.97 vs.439.88 ±23.68,all P =0.000).There was no statistical significance of IFABP levels between abdominal infection group and non-abdominal infection group,the survival group and the death group,or among different AGI-grade groups.The correlation analysis showed that IFABP was statistically related with IL-6 (r=0.794,P=0.000),TNF-α (r=0.878,P=0.010),APACHE Ⅱ score (r=0.428,P=0.000) in patients with severe sepsis.Significant correlations were also found between IFABP and IL-6 (r=0.812,P=0.000),TNF-α (r=0.885,P=0.000) in abdominal infection group,as well as in non-abdominal infection group (IL-6:r=0.739,P=0.000; TNF-α:r=0.828,P=0.000).As shown by multivariate logistic regression analysis,SOFA scores on days 0,1,3 were the independent risk factors for death [odds ratio (OR) was 1.624 (P=0.004),1.411 (P=0.027),1.740 (P=0.012),respectively],but IFABP level,AGI grade,and APACHE Ⅱ score had no influence on death rate.Conclusion IFABP concentrations in patients with severe sepsis were significantly increased,and it is correlated well to IL-6,TNF-α and APACHE Ⅱ score,but did not related obviously with AGI grade and the prognosis of the patients.

Objective To investigate whether heparin has a beneficial effect on lipopolysaccharide(LPS)-induced acute lung injury(ALI)in rats,and to explore the possible underlying mechanisms. Methods Thirty-two adult Sprague-Dawley(SD)rats were randomly assigned into the control,heparin control,model,and heparin treatment groups,with 8 in each group. ALI rat model was reproduced by intratracheal instillation of LPS at a dose of 1 mg/kg. The rats in the control and heparin control groups received an equal volume of normal saline at the same times. The rats in the heparin control and heparin treatment groups were intravenously received 50 U/kg heparin every 1 hour after the induction of ALI. Animals were sacrificed 24 hours after LPS challenge. Bronchoalveolar lavage fluid(BALF) and lung tissue samples were collected. Histopathological evaluation,lung wet/dry(W/D)ratio,malondialdehyde (MDA),nitric oxide(NO)and myeloperoxidase(MPO)were analyzed. Enzyme-linked immunosorbent assay(ELISA) was used to measure the concentration of inflammatory factor in BALF. Expression of inducible nitric oxide synthase (iNOS)mRNA in the lung of rats was measured by reverse transcription-polymerase chain reaction(RT-PCR). Western Blot was used to determine the expression of transforming growth factor-β1(TGF-β1)and phosphorylation of Smad in the lung tissues. The expression of iNOS in lung was determined by immunohistochemistry. Results In the control and heparin control groups,lung tissue showed a normal structure and clear pulmonary alveoli under a light microscope. In the model group,ALI characters such as extensive thickening of the alveolar wall,significant infiltration of inflammatory cells,demolished structure of pulmonary alveoli,and hemorrhage were found. In the heparin treatment group,heparin treatment markedly alleviated LPS-induced these pathological changes in lung. Compared with control and heparin control groups,lung W/D ratio,lung MDA,NO and MPO levels,and tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6)in BALF in the model group were increased significantly. Compared with the model group, lung W/D ratio,lung MDA,NO and MPO levels,and TNF-αand IL-6 in BALF in the heparin treatment group were significantly decreased〔W/D ratio:7.54±0.17 vs. 10.69±0.15,MDA(mmol/mg):2.01±0.30 vs. 2.51±0.25,NO (μmol/L):3.07±0.21 vs. 3.89±0.14,MPO(U/g):1.94±0.09 vs. 2.74±0.20,TNF-α(μg/L):201.80±0.27 vs. 297.53±0.34,IL-6(μg/L):38.41±0.25 vs. 46.31±0.31,all P<0.05〕. RT-PCR showed that the expression of iNOS mRNA in the heparin treatment group was significantly lower than that in the model group(2-ΔΔCt:3.04±0.18 vs. 4.37±0.15,P<0.05). Western Blot showed that compared with control group,the protein expressions of iNOS and TGF-β1,and phosphorylation of Smad2 and Smad3 were significantly increased,and the heparin could inhibit the protein expressions compared with model group. Immunohistochemistry showed that positive expressions of iNOS in alveolar epithelial cell and capillary endothelial cell in the heparin treatment group were significantly lower than those in the model group. Conclusion Heparin significantly ameliorated the lung injury induced by LPS in rats via the inhibition of nitric oxide synthase expression and the TGF-β/Smad pathway.

BACKGROUND:Chondrocytes may dedifferentiate when they are cultured in vitro,and the capacity of synthetizing glycosaminoglycan (GAG)is also reduced,how to delay the dedifferentiation of chondrocytes is a crucial topic in the field of tissue engineering.OBJECTIVE:To observe the performance of chondrocytes synthetizing GAG at different inoculum densities.DESIGN,TIME AND SETTING:A controlled cellular experiment was performed at the laboratory of Department of Orthopaedics in the Second Hospital of Shanxi Medical University between January 2007 and May 2007.MATERIALS:Five New Zealand rabbits of one month old were used in this study.METHODS:Articular chondrocytes were isolated from both knees and digested using 0.4% pronase enzyme and 0.025% Ⅱ type collagenase.The chondrocytes harvested from the same rabbit were divided into two sets,one was seeded at a constant density of 2×104/cm2 in primary and subculture,the other was cultured at a reduced density of 2×103/cm2 following cellular adhesion.Cellular morphology and proliferation were observed under inverted microscope.The culture media were renewed after the primary cells and passage 1 cells were confluent.MAIN OUTCOME MEASURES:GAG concentration was determined using the modified precipitation method with Alcian blue at 12,24,36,48 and 60 hours following the renewal of culture media.RESULTS:Articular chondrocytes in the primary high-density culture group were polygonal with clear boundaries,they have shown to form colony at 3-4 days.Cells around colonies were more slender than those in the center of colonies,shaping as long polygon.There was no obvious change observed in the morphology of passage 1 cells.In the low-density culture group,cells scattered at early stage and formed colonies at 7 days,cellular morphology showed no significant differences in comparison with high-density culture group.The time of primary cells becoming confluent in the low-density culture group was prolonged compared with high-density culture group.The GAG concentration in supernatants in the primary cells of low-density culture group was significantly lower than that in primary cells and passage 1 cells of high-density culture group (P＜0.001,P＜0.05).The GAG concentration showed a greater difference along with the prolonging of culture time.CONCLUSION:High-density culture is better then low-density culture to enhance the performance of chondrocytes synthetizing GAG and to retard the velocity of chondrocytes dedifferentiation,which suggests high-density culture contributes to maintain the chondrocytes phenotype and can be considered as a good way of plate culture.

Objective: To establish an evaluation index system for health education of hand,foot and mouth disease（HFMD）in nursery children by Delphi method. Methods: After referring to the relevant literature,an initial health education index system for HFMD of nursery children was established,including four first-level indicators,twelve second-level indicators and forty-six third-level indicators. Two rounds of expert consultation were conducted according to Delphi method. The enthusiasm of experts was evaluated by response rate,and the authority of experts was evaluated by authority and variation coefficient. The consultation questionnaire in the second round was based on the results of the first round and was scored again in the same way. Then the evaluation index system of HFMD health education for nursery children was finally determined. Results: Thirteen experts participated in two rounds of consultation,including four aged 40-49 years and nine aged 50-59 years;two of deputy senior title and eleven of senior title;one worked in the health administration department,six in the CDC,five in health education institutions and one in kindergarten. The two rounds of expert consultation were carried out effectively within the time set and the response rate reached 100%. In the first round,the judgment coefficient,familiarity coefficient and authority coefficient were 0.92±0.06,0.85±0.12 and 0.88±0.06,respectively. One first-level indicator,two second-level indicators and twelve third-level indicators were added,seven third-level indicators were deleted,and the contents of one second-level indicator and ten third-level indicators were improved in the first round. In the second round,the judgment coefficient,familiarity coefficient and authority coefficient were 0.95±0.07,0.88±0.10 and 0.91±0.06,respectively. The coordination coefficients of the first-,second- and third-level indicators in the second round were 0.170,0.166 and 0.283,respectively,and the coefficients of variation were all less than 0.25. After two rounds of discussion,five first-level indicators,fourteen second-level indicators and fifty-two third-level indicators were finally established as the evaluation index system of HFMD health education for nursery children. Conclusion The evaluation index system of HFMD health education for nursery children established by Delphi method has high authority and practicability,and it can be used to comprehensively evaluate the effects of HFMD health education on nursery children.

Objective: To observe the role of Qudu medicinal granules (祛毒冲剂) on enterogenous endotoxemia . Methods: Sixty-one cases with enterogenous endotoxemia were randomly divided into two groups:Qudu medicinal granule group (n=30) that was treated with Qudu medicinal granules combined with western medicine, and smecta group which was treated with smecta and western medicine (n=31). Changes of symptoms and signs were observed before treatment and 1, 3, 7 days after treatment. Blood samples were collected in the morning to measure the white blood cell (WBC), plasma lipopolysaccharide (LPS) and tumor necrosis factor (TNF) levels. Results: Recovery speed of WBC count in Qudu medicinal granule group was faster than that of the smecta group, there was significant difference on the third day after treatment (P

BACKGROUND: The direct ratio is formed between the size of osseous defect around implant and the required time of complete repair. It is viewed that bone implantation is suggested if osseous defect is larger than 1 mm, which benefits the bone regeneration and early fixation of implant.OBJECTIVE: To compare the effects between coralline granules and hydroxyapatite (HA) during union after immediate implantation.DESIGN: Group observation and controlled experiment was designed.SETTING: Department of Stomatology Renmin Hospital of Wuhan University.MATERIALS: HA coated implant, HA granule, coralline granule and 3adult mongrel dogs.METHODS: The experiment was performed in Department of Stomatology, Renmin Hospital of Wuhan University from August 2002 to April 2003.Under anesthesia, 6 foramens were drilled on femurs of 3 dogs (3 foramens on each side) to result in osseous defect. Coralline granules were embedded in the osseous defect around the implant in all of proximal ends (coralline granule group, CG group), HA granules were embedded in the osseous defect around the implant in all of distal ends (HA group) and nothing was embedded in the defect around the implant in the center (the control). One dog was sacrificed under anesthesia on the 2nd, 3rd and 4th months after operation successively and the samples were collected from the implanted section in each group for X-ray examination and scanning electronic microscopic observation.MAIN OUTCOME MEASURES: X-ray examination on implanted sections and scanning electronic microscopic observation on samples in each the 4th month, it was observed that implants and osseous tissue were closely integrated in CG group and HA group and there was still partial osseous on samples in each group: on the 4th month, it was discovered that the regenerated osseous tissue was matured completely in CG group and few coralline granule residue was left. In HA group, the regenerated osseous tissue was matured completely, but there was still a large amount of HA granules left and the granules had not been absorbed obviously. In the control group, the space appeared partially in the implanted cervical region.CONCLUSION: The implantation of artificial bone is suggested if the osseous defect around immediate implant is larger than 1 mm. As the implanting materials, coralline granules and HA granules act on the most advantageous guide activity. Coralline granules can be degraded and substituted with osseous tissue,but HA granules cannot be absorbed, which affects osseous reconstruction.