Objective To determine the ability of biofilm formation of Staphylococcus ephtermidis isolates and analyze the correlation between the icaA gene and its expression and biofilm formation. Methods Collecting 205 Staphylococcus epidermidis isolates identified with normal laboratory tests (coagulase-negative, biochemical identification, polymyxin-resistant and novobiocin-sensitive ), the suspected isolates were con-formed with API-Staph. Biofilm production was assessed by incubating the strains on Congo Red Agar (CRA) plates and quantitative biofilm production determined by a 96-well tissue culture plate and biofilm morphous were detected by scanning electron microscope ( SEM ) ; Amplifying partial fragments of icaA genes with PCR; Analyzing the expression levels of icaA gene with RT-PCR through Bio-Rad system and Quantity One software. Results 24 isolates showed positive in CRA tests, 22 isolates were positive in semiquantita- tive adhesion assays and 28 isolates existed icaA gene among 205 isolates of Staphylococcus epidermidis. The icaA-positive strains demonstrated biofilm formation (microcolonies on silica films ) while icaA-negative strains only adhered as individual cells under scanning electron microscope. All 22 strains which showed positive in semiquantitative adhesion assays harbored the icaA gene. The expression levels of icaA gene with RT-PCR in 6 Staphylococcus epidermidis isolates showed a higher tendency in 4 strains which demonstrated positive in semiquantitative adhesion assays than 2 negative strains in semiquantitative adhesion assays. Conclusion The isolations of Staphylococcus epidermidis have the abilities of forming biofilm, and the icaA gene and its normal expression is the important molecular biology foundation of biofilm formation. Other fac-tors maybe involve in the expression of icaA gene in Staphylococcus epidermidis isolates.

ObjectiveTo determine the ability of biofilm formation of Staphylococcus epidermidis isolates and study the influence of different extracellular DNA(eDNA) levels in S.epidermidis isolates on the ability of biofilm formation.MethodsDetect the biofilm-formation ability of 227 S.epidermidis isolates with adhesion assays,amplify the icaA gene fragment with PCR.The S.epidermidis isolates were divided into biofilm formation(BF) group and non-biofilm formation (NBF) group according to adhesion assays and icaA amplification.Detect eDNA levels of S.epidermidis in planktonic culture and microtitre plate static culture.The eDNA in S.epidermidis biofilms stained by AO-PI was observed by CLSM.Results26 isolates were positive in adhesion assays and 32 isolates existed icaA gene among 227 S.epidermidis isolates.Select 20 isolates with positive adhesion assays and positive icaA amplification for BF group.Select 19 isolates with negative adhesion assays and negative icaA amplification for NBF group.The eDNA levels were (32.2±10.1)μg/ml,(33.6±11.9) μg/ml,(34.3±10.0) μg/ml in BF group when cultured in planktonic condition for 2,4,6 h,while the eDNA levels in NBF group were (28.7±8.9) μg/ml,(31.5±11.7) μg/ml,(31.8±12.7) μg/ml respectively.There were no significant differences between the two groups for these three phases(P＞0.05),though the eDNA levels of BF group were higher than that of NBF group.The eDNA levels were (740.0±264.4) ng/A600 in BF group when cultured in static microtitre plate,higher than that of NBF group,(80.1 ±31.1) ng/A600,and the difference between these two groups was significant.The eDNA in BF isolate Y36 biofilms could be visualized by staining with AO and PI when observed by CLSM,while neither biofilm structure nor eDNA appeard when NBF isolate Y26 was cultured for 24 h.ConclusionS.epidermidis isolates have the ability of biofilm formation.eDNA is one of the important matrix components in the S.epidermidis biofilm-forming process.The eDNA of static culture in microtitre plate was more efficient than planktonic culture in the case of estimating the ability of biofilm formation of S.epidermidis.

Objective To investigate the relationships between the expressions of cidA and lrgA and the biofilm formation of Staphylococcus epidermidis clinical isolates. Methods Thirty-nine S. epidermidis clinical isolates were divided into biofilm formation group and non-biofilm formation group according to adhesion assays and icaA amplification, 20 strains for biofilm formation group and 19 strains for another group respectively. Amplify cidA and lrgA fragment with PCR, detect mRNA levels of cidA and lrgA with RT-PCR and then calculate the ratios of cid/lrg mRNA. Results All of the 39 isolates of S. epidermidis existed cidA and lrgA gene.Select 2 strains of S. epidermidis, Y36 and Y26, at random for detecting the cidA and lrgA mRNA levels of different phases, the highest level of cidA mRNA shows on 4 h's culture while lrgA shows on6 h. The relative expression levels of cidA of 39 isolates of S. epidermidis were 0.340 + 0.250 in biofilm formation group and 0. 406 +0. 408( P =0. 541 ) in non-biofilm formation group for 4 h culture, while the relative expression levels of lrgA were 0.325 ± 0.218 and 0.253 ± 0.211 respectively ( P = 0. 299). There were no significant differences between these two groups. The ratios of cid/lrg mRNA were 1.067 ±0.529 and 1.958 ±1.877 respectively, the difference of population distribution was significant( P = 0.001 ). Conclusion The expressions of cidA and lrgA of S. epidermidis clinical isolates may be time-limited. There were no significant differeces of expressions of cidA and lrgA between biofilm formation group and non-biofilm formation group. The ratios of cid/lrg mRNA maybe have some biological significance.

Electron microscopic changes about the effect of gossypol acetic acid ongastrocnemius muscles of male monkeys,rats and guinea pigs were studied.The resultsare as follows:The ultrastructure of gastrocnemius muscles of adult male Rhesus monkeys afteradministration of gossypol orally with a daily dose of 4mg/kg for 2 years showedapparently normal in appearance,except some nuclear condensation and dissociationof nuclear membrance seen in individual animal.After oral aministration of gossypol in adult male Wistar rats with an antifertilitydosage,i.e 20mg or 30mg/kg/day for 30 days,no obvious difference was seen betweenthe experimented and control groups.Except mild distension of the smooth endoplasmicreticulum and dissociation of nuclear membrane sometimes seen in some of theexperiment animals.Similar results was obtained in the male guinea pigs.After a heavy dose(60mg/kg/day)of gossypol given to rats for 37 days,mostof the myofibrils still keep their normal periodic structures,each fiber is maintainingtheir transversly arranged pattern of dark A-bands and light I-bands alternativelyalong the whole length of the fiber.But some myofibrils had lost their normal striation.The width of the I-band increased and the breaking down or lysis precessesin few or even an individual myofibril had begun.There were no obvious pathologicalchanges seen in the mitochondria.Persistent oral administration of gossypol in ratewith a dosage of(60mg/kg/day)for 55 days,the pathological changes became moreobvious.The mitochondria became disordered in arrangement.Concomitant with thesschanges,the myofibrils appeared dystropic and fragmented.It indicated the presenceof myofibrillar degeneration.If the daily dose are increased to 60mg/kg for 25 days in guinea pigs.Theremight be no significant pathological changes in the myofibrils in them.Nevertheless,some mitochondria were swollen in various extents and the cristae became fragmentedor dissolved,which was known as mitochondrial necrosis.The relationship between hypokalemic paralysis and the present results inducedby toxic effect of gossypol were analysed and discussed.

Objective To determine the clinical value of comcombined detection of serum FT 3 ,FT4 ,TSH ,TRAb ,TPOAb on outcome of women who have hyperthyroidism during pregnancy .Methods 100 patients with hyperthyroidism during pregnancy and 102 healthy pregnant women was selected in this research .Results The difference between the disease group and control group was statistically significant(P<0 .05) .Conclusion The combined detection of serum FT3 ,FT4 ,TSH ,TRAb ,TPOAb may have special impact on outcome of pregnant women .

Objective To analyze the correlation between the expression of Abl interactor 1 (ABI1) and the clinicopathologic characteristics in colorectal adenocarcinoma tissues.Methods Immunohistochemistry was used to determine ABI1 expression in human colorectal adenocarcinoma tissues and matched adjacent tissues.A statistical analysis was used to determine the potential correlation between ABI1 expression and clinicopathological characteristics and prognosis.Results ABI1 is up-regulated in colorectal adenocarcinoma tissues compared with matched adjacent tissues (P =0.000),ABI1 expression is significantly correlated with infiltration (P =0.043) and differentiation (P =0.040),but not with sex,age,tumor location,clinical stage,lymph node metastasis and distant metastasis (P ＞0.05);It was also shown that ABI1 expression had a significant influence on prognosis (x2 =11.090,P =0.001).Multivariate analyses showed that high ABI1 expression is not an independent poor prognostic factor for overall survival (P =0.119).Conclusion ABI1 overexpression might act as pro-oncogene for patients with colorectal adenocarcinoma.

Objective:To study the factors influencing the happiness of dementia patients and their caregivers,provide guidance for improving their well-being.Methods:A total of 94 pairs of patients and their caregivers who were admitted to the neurology department of Peking Union Medical College Hospital from April 2015 to April 2016 were selected, the demographics of each patient and their caregivers were recorded. The Mini-Mental State Examination(MMSE) of patients with dementia, Role of Overload Scale(ROS) of caregivers, Dyadic Relationship Strain(DRS), Quality of Life for Dementia(QOL-D), Self-Evaluation Scale-Depression(CES-D) were recorded. Layered linear model was used to make regression analysis between the influencing factors and the scores of QOL-D and CES-D.Results:The results of the multi-layer linear model of uncontrolled variables in the fixed effect model: the results of QOL-D suggested that the score of patients with dementia was β 1j= 31.01±0.77, and the score of caregivers was β 2j= 35.15±0.88; the results of CES-D suggested that the scores of dementia patients and caregivers were β 1j = 14.55 ± 1.03 and β 2j = 13.11 ± 1.44, respectively. The random effects model suggested that there were statistical differences in the heterogeneity of the QOL-D score and the CES-D score variance component for dementia patients and caregivers (χ 2 values were 98.94-168.06, P<0.01). It indicated that the data was heterogeneous, adjusting the level 2 model, and the final results in the adjusted regression analysis suggested: caregiver relationship pressure (DRS), dementia patient self-awareness assessment (MMSE), caregiver care-related stress (ROS), dementia patient relationship stress (DRS) significantly affected the quality of life score (QOL-D) in both well-being ( β values were -3.22-0.43, P<0.05). Dementia patient relationship stress (DRS), caregiver-related stress (ROS), and caregiver relationship stress (DRS) significantly affected depressive symptoms in both well-being ( β values were 5.34, 3.26, 1.62, P<0.05). Conclusions:A comprehensive assessment of dementia patients and caregivers is needed. The combined family relationship is tense and the pressure associated with caregivers needs to be psychologically counseled.

Objective To study the relationship of Helicobacter pylori(Hp)infection degree with serum pepsinogen(PG)and the tumor markers related with gastric cancer .Methods Totally 342 cases of health physical check‐up in our hospital from January to June 2015 were selected .13 C‐UBT was performed to evaluate the Hp infection and infection severitys .The level of serum PG was detected by ELSIA and the levels of tumor markers were detected by luminescence immunoassay .Then the results were statistically analyzed with the SPSS statistical software .Results The positive rate of Hp in 342 research subjects was 49 .42% ,and there was no difference between the male and the female groups(P>0 .05) .The level of serum pepsinogen Ⅰ (PGⅠ )and pepsinogen Ⅱ (PGⅡ)in the Hp(+ + )and Hp(+ + + )groups was significantly higher than that in the Hp negative group ,while the PGⅠ /PGⅡ lev‐el was significantly lower than that in the negative group (P<0 .05) .The level of carbohydrate antigen 724(CA724)and carcino‐em‐bryonic antigen(CEA)had the statistical difference between the Hp(+ + + ) group and the Hp negative group(P= 0 .040 ,P=0 .010) .The Pearson correlation analysis displayed that Hp infection was related with PG Ⅰ ,PGⅡ ,PGⅠ /PGⅡ ,CA724 and CEA . There was a positive correlation between PG Ⅱ and CA50)(r=0 .116 ,P=0 .032);there was a negative correlation between PG Ⅰ /PGⅡ and CA50(r= -0 .193 ,P=0 .000) .Conclusion The combined detection of Hp ,PG and tumor markers could be used as one of methods for screening benign and malignant gastric diseases in the healthy physical check‐up population ,which has an important value for the prevention and intervention of related disease occurrence and development .

BACKGROUND:Accumulative evidence supports that co-culture technology can be applied to construct the tissue-engineered cartilage with excellent biological characters. OBJECTIVE:To elaborate the co-culture concept and conclude and analyze seed cell sources, cel mixed ratio, spatial y-defined co-culture models and biomaterials in co-culture systems to conclude and analyze the biological characters of tissue-engineered cartilage, and to prospect progression of co-culture systems in cartilage tissue engineering. METHODS:The first author retrieved the databases of PubMed, Web of Science, and CNKI for relative papers published from January 1976 to May 2016 using the keywords ofco-culture, co-culture systems;articular cartilage, chondrocytes, mesenchymal stem cells;tissue engineering, articular cartilage tissue engineeringin English and Chinese, respectively. Finally 60 literatures were included in result analysis, including 1 Chinese and 59 English articles. RESULTS AND CONCLUSION:Co-culture technology emphasizes the role of microenvironment in terms of various physical, chemical and biological factors in the cell processing. In cartilage tissue engineering, co-culture systems contribute to maintain the viability and natural cell phenotype of chondrocytes and induce cartilage differentiation of mesenchymal stem cells. In addition, co-culture technology provides a novel way for cartilage tissue engineering to overcome the shortage of chondrocytes and repair injury to the cartilage-subchondral bone. However, the mechanisms of cell-cell interaction in co-culture systems still need to be explored in depth, so as to optimize the co-culturing conditions and construct perfect tissue-engineered cartilage.

Objective To establish a mouse model of biliogenic severe acute pancreatitis (SAP) by using a self-made device for retrograde injection of sodium taurocholate into common bile duct,and to investigate the improvement of the device on retrograde injection of sodium taurocholate into common bile duct and its safety.Methods Thirty-six adult male ICR mice were randomly divided into biliogenic SAP model group and sham group,with 18 mice in each group.A 40 U disposable insulin syringe,a 200 μL tips and a 25 μL micro-syringer were used as basic materials for making the mouse common bile duct injection device [National Utility Model Patent (ZL 2014 2 0694365.4)].In model group,3.5％ sodium taurocholate (1 mL/kg) was injected retrogradely into the common bile duct of mice,whilst in sham group,the mice underwent the injection of equal amount of normal saline instead.Six mice in each group were sacrificed at 6,24 and 48 hours after operation,and the abdominal aortic blood was collected.Serum amylase (AMY),alanine aminotransferase (ALT),creatine kinase-MB (CK-MB),serum creatinine (SCr),oxygenation index (PaO2/FiO2) as well as serum Ca2+ were.determined.Pathological change in pancreas was observed under conventional light microscopy after hematoxylin and eosiu (HE) staining,and the impairment was evaluated by a widely used score system.Results The injection device was easily placed into mouse common bile duct under macroscopic observation.Six hours after operation,the levels of serum AMY,ALT and SCr in model group were significantly higher than those in sham group,and peaked at 24 hours,and they slightly decreased at 48 hours,which were still significantly higher than those of the sham group [24-hour AMY (U/L):7 325 ± 1 154 vs.1 737 ± 197,24-hour ALT (U/L):176.0±5.0 vs.38.3 ± 2.0,24-hour SCr (tmol/L):46.3 ± 1.5 vs.17.8 ±0.6,all P ＜ 0.01].The level of CK-MB at 6 hours in the model group was significantly higher than that of the sham group,and peaked at 48 hours (U/L:749.8±42.2 vs.383.3±35.5 at 6 hours,3 340.1 ± 203.6 vs.704.6 ± 63.5 at 48 hours,both P ＜ 0.01).PaO2/FiO2 at 6 hours after the operation in model group was significantly lower than that of sham group,then it began to rise at the similar level in sham group at 48 hours [mmHg (1 mmHg =0.133 kPa):327.5±33.8 vs.424.8±31.0 at 6 hours,P ＜ 0.01;429.8 ±41.8 vs.464.7±43.3 at 48 hours,P ＞ 0.05].Ca2+ level in model group was continuously decreased after operation,and it was significantly lower than that of sham group at 48 hours (mmol/L:1.58 ± 0.14 vs.2.45 ± 0.21,P ＜ 0.01).The pancreatic edema was obvious after operation in sham group,with the observation time prolongation,the changes were gradually improved;pancreatic focal necrosis was found at 6 hours after operation in model group,and it was secondary aggravated,and pancreatic lobule structure disappearance and inflammatory cells extensive infiltration was found at 48 hours.Pathological score of the model group was significantly higher than that of sham group at each time point,and peaked at 48 hours (13.3 ±0.3 vs.3.0±0.1,P ＜ 0.01).Conclusion It is a highly efficient and low-cost way to induce biliogenic SAP in mice by retrograde injection of 3.5％ sodium deoxycholate into common bile duct via the self-made injection device,and the model conformed to the clinical characteristics of biliogenic SAP.