Gut microbial metabolite trimethylamine-N-oxide (TMAO) is associated with type 2 diabetes (T2DM). Decreased insulin sensitivity is a significant etiological factor of T2DM. Adipocytes, myocytes, and hepatocytes are the three major target cells for insulin. This study aims to investigate the effects and mechanisms of TMAO on the insulin sensitivity of these target cells. Research results indicate that in different ages of db/db diabetic mice, plasma TMAO levels were increased. TMAO significantly inhibits the insulin signaling pathways in these three major insulin target cells, reduces glucose uptake in 3T3-L1 adipocytes and L6 myocytes and downregulates genes related to gluconeogenesis in primary mouse hepatocytes. Furthermore, in mice with normal insulin sensitivity, elevating plasma TMAO levels to those seen in db/db mice using a minipump results in impaired glucose tolerance and hyperinsulinemia. All animal experiments were carried out with approval of the Experimental Animal Welfare Ethics Committee of the Institute of Materia Medica (Chinese Academy of Medical Sciences and Peking Union Medical College). Mechanistic studies suggest that TMAO exposure increases the levels of endoplasmic reticulum stress-related proteins in these three major insulin target cells. In summary, TMAO directly attenuates insulin sensitivity in insulin target cells, and its mechanism of action may involve enhancing endoplasmic reticulum stress.

Objective To investigate the methods of isolation and culture in vitro and detect the surface markers of human umbilical cord mesenchymal stem cells.Methods Human umbilical cord Wharton' s jelly was separated and cut up as small as possible,and then cultured with α-MEM.Human umbilical cord mesenchymal stem cells could be obtained by culturing the tissue block adhered the bottle wall.And the cells were passaged at a certain density.The surface markers of human umbilical cord mesenchymal stem cells were detected by FACS when the cells were in Generation Three.Results Human umbilical cord mesenchymal stem cells were obtained from Wharton' s jelly conveniently,with fibroblast shape and stable proliferation and passage.CD29,CD44,CD105 were strongly expressed on human umbilical cord mesenchymal stem cells.But CD45,CD34,HLA-DR,HLA-G,CD80,CDs6 were not expressed.Conclusion Human umbilical cord mesenchymal stem cells can be obtained effectively from the culture of the tissue block,which provides a rich source of cells for tissue engineering.

Objective To investigate the distribution characteristics of serum small dense low density lipoprotein-cholesterol (sdLDL-C) levels in healthy normolipidemia and hyperlipidemia and analyze the correlation between sdLDL-C and other serum lipids.Methods Totally 1012 normolipidemic subjects (18 93 years old,503 male,509 females) were grouped according to gender and age (18 -29,30 -69 and ≥70 years old).And 433 hyperlipidemic subjects (23 -90 years) were divided into the following 3 groups based on fasting triglyceride (TG) and low density lipoprotein-cholesterol (LDL-C)levels:hypertriglyceridemia (n =165 ),high-LDL-C ( n =129) and combined hyperlipidemia ( n =139 ).The sdLDL-C and other serum lipids were measured by Olympus AU2700 automatic analyzer.Analysis of Variance and Kruskal-Wallis H test was used for statistical analysis.Results The distribution of the sdLDL-C levels in normolipidemic subjects was near normal distribution.The sdLDL-C levels had differences in gender and age.In the 18 -29 and 30 -69 years old group,the mean values of sdLDL-C were significantly higher in males than in females [ (0.55 -0.21 ) mmol/L vs (0.47 ±0.22) mmol/L,t =2.212,P =0.028 and (0.66±0.28) mmol/L vs (0.62±0.25) mmol/L,t =2.121,P=0.034].In the ≥70 years old group,the difference of sdLDL-C levels in gender was not statistically significant [ male ( 0.54 ± 0.21 )mmol/L vs female (0.54 ± 0.22 ) mmol/L,t =0.022,P =0.982] ; the mean value was ( 0.54 ± 0.22 ) mmol/L The hyperlipidemic subjects had conspicuous higher levels of sdLDL-C compared with normolipidemia [ ( 1.25 ±0.44) mmol/L vs (0.60 ±0.26) mmol/L,t =29.306,P ＜0.001 ].Among all of groups,the combined hyperlipidemia group had the highest sdLDL-C level [ ( 1.52 ± 0.49) mmol/L,F =525.66,P ＜0.001 ] ; the hypertriglyceridemia group had the highest sdLDL-C/LDL -C level (0.47 ±0.12,F =287.93,P ＜0.001 ) and the high-LDL-C group had the highest level of non-sdLDL-C [LDL-C subtract sdLDL-C,(2.71 ± 0.52) mmol/L.F =336.32,P ＜ 0.001 ].The sdLDL-C showed a good positive correlation with TC,TG,LDL-C,ApoB and BMI ( rs =0.66,0.68,0.65,0.79 and 0.27,P ＜ 0.001 ),and negative correlation with HDL-C and ApoA1 ( rt =- 0.42 and - 0.37,P ＜ 0.001 ).Based on partial correlative analysis,sdLDL-C showed a different correlation with TG,LDL-C and TC ( r =0.42,0.28 and 0.15,P ＜ 0.001 ).Conclusions LDL-C and TG are the important factors influencing the sdLDL-C levels.However,TG has greater effect than LDL-C.The sdLDL-C is an appropriate and good index to evaluate the small dense low density lipoprotein (sdLDL) mass and the overall situation of lipid metabolism.In order to make full use of sdLDL in the clinical treatment and health assessment,it is necessary to establish sdLDL-C reference intervals through the survey of distribution of sdLDL-C levels in different geographic areas.

Objective To study the serum small dense low density lipoprotein-cholesterol (sdLDL-C)concentration in patients with clinical and sub-clinical primary hypothyroidism,and to explore the clinical significance of serum sdLDL-C determination in pa-tients with hypothyroidism.Methods 60 patients with clinical hypothyroidism(clinical hypothyroidism group),96 cases with sub-clinical hypothyroidism(sub-clinical hypothyroidism group)and 132 normal subjects (normal group)were enrolled in the study. FT3,FT4,thyroid stimulating hormone (TSH),TG,TC,HDL-C,LDL-C,apolipoprotein AⅠ(ApoAⅠ),apolipoprotein B (ApoB), and sdLDL-C concentrations were measured.Analysis of Variance,Kruskal-Wallis H test and Spearman rank correlation analysis were used for statistical analysis.Results The concentration of sdLDL-C in sub-clinical hypothyroidism group was significantly higher than that in normal group (t=5.78,P <0.01),the concentration of sdLDL-C in clinical hypothyroidism group was signifi-cantly higher than that in sub-clinical hypothyroidism group(t=-2.88,P <0.01)and in normal group (t=6.68,P <0.01).There was statistically significant difference among groups (F =37.66,P <0.01).Correlation analysis showed that sdLDL-C in primary hypothyroidism patients was positively correlated with LgTSH (r =0.203,P =0.011 ),and negatively correlated with FT4 (r =-0.169,P =0.035).Conclusion The high concentration of sdLDL-C is a manifestation of dyslipidemia in primary hypothyroidism patients,and an important risk factor for atherosclerosis in primary hypothyroidism patients.

OBJECTIVE: To study the effect of continuous renal replacement therapy on acute renal failure and multiple organ dysfunction syndrome following simultaneous pancreas-kidney transplantation. METHODS: A patient was complicated with acute renal failure, severe acute pancreatitis, lung infection, bleeding in anastomosisbetween duodenum and jejunum, and peritonitis following simultaneous pancreas-kidney transplantation. He was treated with immunosuppressor, antibiotics, amylopsin inhibitor, haemostatic and alimentation; at the same time, he was treated with continuous renal replacement therapy for 22 days. The Baxter system was used for continuous venovenous hemofiltration. RESULTS: The vital signs and hemodynamic indicators were stable during continuous renal replacement therapy. Pulmonary edema was well controlled, and acid-base equilibrium of water electrolyte was maintained. The function of vital organs was stableand graft function was normal following continuous renal replacement therapy for 22 days. He was completely cured and out of hospital on day 40. CONCLUSION: Continuous renal replacement therapy plays an important role in treating acute renal failure and multiple organ dysfunction syndrome following simultaneous pancreas-kidney transplantation. Thus, it is a well kidney support for ultaneous pancreas-kidney transplantation.

To assess the impact of the operational simulation training system on the cultivation of ophthalmic students.
●METHODS: Four hundred and eighty ophthalmic students in China Medical University from 2009 to 2011 were chosen as objects and divided into two groups at random. The students in the experimental group used operation simulation training system combined with traditional teaching methods, while the students in the control group only used traditional teaching methods. We examined the operation skills and the confidence of all the students half a year later.
●RESULTS: The improvement of the operation skills during the suture technique examination, and the confidence for operation technology were significantly different in two groups. The basic skills of the students in the examination groups improved apparently during the operational simulation training and they were full of confidence to their operational technique.
●CONCLUSlON: The operational simulation training is useful for promoting the clinical ability for ophthalmic students and should be widely popularized.

To understand the structure of the soluble region of Enterovirus 68 3A protein, we construct a prokaryotic expression vector expressing the soluble region of EV-D68 3A protein, and identify the forms of expression product after purification. The EV-D68 3A(1-61) gene was amplified by PCR and then cloned into the expression vector pET-28a-His-SUMO. The recombinant plasmid was transformed into Escherichia coli BL21 induced by IPTG to express the fusion protein His-SUMO-3A(1-61). The recombinant protein was purified by Ni-NTA Agarose and cleaved by ULP Protease to remove His-SUMO tag. After that, the target protein 3A(1-61) was purified by a series of purification methods such as Ni-NTA, anion exchange chromatography and gel filtration chromato- graphy. Chemical cross-linking reaction assay was taken to determine the multiple polymerization state of the 3A soluble region. A prokaryotic expression vector pET28a-His-SUMO-3A(1-61) expressing the solution region of EV-D68 3A was successfully constructed and plenty of highly pure target proteins were obtained by multiple purification steps . The total protein amount was about 5 mg obtained from 1L Escherichia coli BL21 with purity > 95%. At the same time, those results determined the homomultimer form of soluble 3A construct. These data demonstrated that the expression and purification system of the soluble region of 3A were successfully set up and provide some basic konwledge for the research about 3A crystal structure and the development of antiviral drugs targeted at 3A to block viral replication.
Amino Acid Sequence ; Enterovirus D, Human ; chemistry ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Models, Molecular ; Molecular Sequence Data ; Protein Structure, Tertiary ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics ; metabolism

Objective To discuss the clinical value of the treatment of intercondylar fractures of the distal femur using open reduction and internal fi xation. Methods From July 1995 to December 2001, a total of 32 intercondylar fra ctures of the distal femur were treated with open reduction and internal fixatio n with a plate, 95? blade plate, a dynamic condylar screw (DCS) or a condyla r buttress plate. There were 13 cases of type-C1, 10 type-C2 and 9 type-C3 ac cording to the AO/ASIF standard. The outcomes of internal fixation were evaluate d according to preoperative and postoperative radiographs and postoperative func tions of the knee. Results 27 patients were followed-up from 8 months to 8 year s. The results were excellent in 14 cases, good in 9, poor in 4 according to San ders standard. Conclusion Open reduction and internal fixation is an ideal ch oice for intercondylar fractures of the distal femur.

Purpose To compare the CTPA image quality by using contrast agent with different concentration at different injection rate so as to provide suitable contrast agent injection for patients.Materials and Methods A total of 346 patients with suspected acute pulmonary embolism who required to undergo CTPA examination were randomly assigned to high (370 mgI/ml) and low (320 mgI/ml) concentration groups,and each group was further divided into six subgroups with different velocity (3.0,3.2,3.4,3.6,3.8 and 4.0 ml/s).The CT value of the main pulmonary artery,right pulmonary upper lobe artery and right lung under leaf posterior basal segmental artery was measured.Results In the high concentration group,there were no significant differences in pulmonary artery average CT value,noise,single to noise ratio (SNR) and contrast to noise ratio (CNR) among the subgroups with different velocity (P＞0.05).In the low concentration group,the difference was not statistically significant in pulmonary artery average CT value (P＞0.05) among the subgroups with different velocity;however,the noise,SNR and CNR of 3.0 ml/s subgroup had significant differences compared with other subgroups (P＜0.05).There was no significant difference in average CT value of pulmonary artery between the subgroups with the same velocity in the two concentration groups (P＞0.05).In addition,except that the noise,SNR and CNR of 3.0 ml/s subgroup showed significant differences with other subgroups either in high concentration group or in low concentration group (P＜0.05),there were no significant differences in the above-mentioned parameters among other subgroups with the same velocity in both groups (P＞0.05).Conclusion Compared with high concentration contrast agent,the image obtained by using low concentration contrast agent shows no difference in pulmonary artery average CT value but with low iodine flow and iodine flow rate,which can reduce the risks of contrast media induced nephropathy (CIN) and contrast agent extravasation.

In the present study, the specifically knockdown models of P-gp or MRP2 were constructed by using a series of chemically synthesized small interfering RNA (siRNA) in vitro. The expression of P-gp and MRP2 was measured by real-time PCR and Western blot, and the function was evaluated by applying P-gp and MRP2 substrate, rhodamine and methotrexate. The results showed that MRP2 siRNA-3 or P-gp siRNA-2 significantly decreased the mRNA expression of MRP2 or P-gp, the inhibition ratio was 68% or 84%; MRP2 siRNA-3 or P-gp siRNA-2 at a dose of 80 nmol x L(-1) significantly reduced the protein expression of MRP2 or P-gp at 48 h after treatment, the inhibition ratio was 62% or 70%. Meanwhile, other transporters were not influenced by siRNA. When pretreatment with MRP2 siRNA-3 or P-gp siRNA-2, the efflux of methotrexate or rhodamine decreased significantly and the intra-cellular concentration increased. The results suggested that chemically synthesized siRNA could significantly inhibit the expression and function of MRP2 and P-gp, and the model of RNAi in vitro could be used to evaluate the role of efflux transporters in transportation of drugs.
ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Gene Knockdown Techniques ; Multidrug Resistance-Associated Proteins ; genetics ; RNA Interference ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction