AIM:To investigate whether specific small interfering RNA ( siRNA) targeting Toll-like receptor 4 ( TLR4) gene inhibits the proliferation of cervical cancer cell line HeLa through NF-κB signaling pathway and down-regula-tion of Bcl-2 expression.METHODS:Three specific TLR4 siRNAs and 1 negative siRNA were transfected respectively in-to HeLa cells.The expression of TLR4 at mRNA and protein levels was measured by reverse transcription-polymerase chain reaction ( RT-PCR) and Western blotting , respectively .The protein expression of p 65 and Bcl-2 was detected by Western blotting.The cell proliferation was determined by MTT assay and plate colony formation assay .The apoptotic rate of the cells and the cell cycle were analyzed by flow cytometry .RESULTS:In comparison with the other 2 kinds of TLR4 siR-NAs, TLR4-siRNA-003 demonstrated the strongest silencing effect on TLR4 gene expression in the HeLa cells at 48 h after transfection.The expression of TLR4 at mRNA and protein levels was reduced by 62% and 89%, respectively.The pro-tein levels of p65 and Bcl-2 significantly decreased .The growth rate of HeLa cells transfected with TLR 4-siRNA-003 was significantly inhibited .Moreover , the cell apoptotic rate increased significantly and the cell cycle was arrested at G 1 phase as the HeLa cells were transfected with TLR 4-siRNA-003 for 48 h.CONCLUSION:Specific TLR4 siRNA effectively in-hibits the expression of TLR4 and inhibits the proliferation of cervical cancer HeLa cells through NF-κB signaling pathway and down-regulation of Bcl-2 expression, indicating that TLR4 may be a new target for the treatment of cervical cancer .

ObjectiveTo determine endometrial thickness, endometrial volumeand, endometrial and subendometrial blood flows, by using transvaginal two-dimensional (2D) and three-dimensional (3D) ultrasound examination and to compare the value of all the parameters in the prediction of endometrial receptivity during in vitro fertilization-embryo transfer (IVF-ET) cycles.MethodsOne hundred and twenty patients undergoing IVF cycles were recruited for this study. Transvaginal 2D and 3D ultrasound examination was performed to determine endometrial thickness, endometrial volume, pulsatility index (PI), resisitance index (RI) , systolic/diastolic ratio (S/D) of endometrial, subendometrial blood flows , vascularization index (VI), flow index (FI) and vascularization flow index (VFI) of endometrial and subendometrial regions by experienced sonographers on the day of human chorionic gonadotropin (HCG) injection. Statistical comparison was carried out by Mann-Whitney test. The receiver operator characteristic (ROC) curve analysis was applied and the area under the curve (AUC) was calculated. Statistic analyses were performed using SPSS 13.0. The two-tailed value of was considered statistically significant.ResultsThere were no significant differences between the pregnant group and non-pregnant group in endometrial thickness, endometrial volume (P>0.05). Patients in the pregnant group had lower PI, RI and S/D of endometrial and subendometrial blood flows than those in the non-pregnant group (Z=-3.811,-3.097,-2.071, P=0.001, 0.001, 0.038), with statisticalsignificant difference. Patients in the pregnant group had higher VI, FI and VFI of endometrial and subendometrial regions than those in the non-pregnant group (Z=-1.970,-2.698,-1.981, P=0.490, 0.007, 0.048;Z=-2.098,-2.090,-2.115,P=0.036, 0.037, 0.034), with statistical significant difference. ConclusionsEndometrial and subendometria blood flows measured by transvaginal 2D and 3D ultrasound can be as a reliable indicator to assess endometrial receptivity and predict the pregnancy outcome. Among all the endometrial and subendometrial blood flow parameters, VI of endometrial and subendometrial are better predicators.

Objective To investigate the changes of cardiac structure and function in patients with hepatolenticular degeneration (HLD).Methods From March 2010 to February 2011,90 HLD patients who did not receive formal treatment (observation group) and 30 healthy people (control group) were collected and analyzed with color Doppler echocardiography in the Institute of Neurology,Affiliated Hospital of Anhui University of Traditional Chinese Medicine.Based on the clinical manifestations,cases were divided into hepatic type group (n =31),brain type group (n =42) and brain-visceral type group (n =17).According to the Child-Pugh classification,patients were divided into Child A group (n =71),Child B group (n =10) and Child C group (n =9).Results (1) The left atrial diameter ((28.00 ± 3.11),(29.62 ± 3.44) mm) and left ventricular diameter ((45.69 ± 3.75),(47.10-± 4.73) mm) of HLD patients in brain-visceral type group and hepatic type group were larger than that of the control group ((24.86 ± 2.63),(41.93 ±3.56) mm;t =3.143,4.761,P=0.018,0.000;t=3.764,5.167,P=0.018,0.000).The left atrial diameter and left ventricular diameter of hepatic type group were significantly larger than the brain type group (26.06 ± 3.68,43.34 ± 3.88;t =3.557,P =0.000;t =3.751,P =0.001).The value of E/A in the hepatic type group (1.57 ± 0.37) was significantly lower than the control group (1.93 ± 0.20;t =-0.352,P =0.006).(2) The left atrial diameter ((31.29 ± 1.70),(34.67 ± 1.97) mm) and left ventricular diameter ((48.29 ± 2.81),(53.67 ± 2.67) mm) of Child B and C groups were significantly larger than the control group ((24.86 ± 2.63),(41.93 ± 3.56) rm;t =6.429,9.810,P =0.000,0.000;t =6.357,10.738,P =0.000,0.000),and the Child A group ((26.42 ± 3.05),(43.89 ± 3.76) rm;t=4.871,8.252,P=0.000,0.000;t =4.399,8.780,P=0.003,0.000).The value of E/A of Child B and C groups (1.58 ± 0.32,1.26 ± 0.39) was lower than that of the control group (t =-0.347,0.662,P=0.020,0.000);At the same time,the value of E/A of Child C group was significantly lower than that of Child A group (1.80 ± 0.33;t =-0.530,P =0.000).Conclusions The HLD patients may have cardiac structural and functional changes,mainly manifested as left atrial and left ventricular enlargement and cardiac diastolic dysfunction,whereas a serious impact has not yet been found in systolic function.And the changes of cardiac structure and function were related to the degree of liver cirrhosis in patients with HLD.

Objective To observe the effects of ultrasound medium frequency electrotherapy on pain and content of enkephalins. Methods The pain threshold of the rat was measured before and after the application of ultrasound medium frequency electrotherapy. The content of met enhephalin(MEK) and leu enkephalin(LEK) in the glandula hypophysis and hypothalamus was also measured. A linear regression analysis based on the pain threshold and the content of enkaphalin was conducted to reveal the pain relieving mechanism of the ultrasound medium frequency electrotherapy. Results The level of MEK and LEK in the glandula hypophysis and hypothalamus was significantly increased after a single use action ultrasound medium frequency electrotherapy as compared with that before the treatment ( P

ObjectiveTo determine whether the β2-agonist, clenbuterol, can improve recovery of locomotor function following spinal cord injury.MethodsThe spinal cords of rats were contused with a weight-drop apparatus similar to the NYU impactor at the lever of T10. Locomotor function was determined with the Basso, Beattie, Bresnahan (BBB) Scale. Following 6 weeks of behavioral evaluation, the spinal cords were received histological examination.ResultsClenbuterol improved the recovery of locomotor function and increased the amount of spared spinal cord tissue.ConclusionClenbuterol caused the enhancement of recovery of locomotor function and spared spinal cord tissue following contusion.

Objective:To investigate the effects of Helicobacter pylori on NLRP3 inflammasomes activation in THP-1 ( human monocytic cell line) -derived macrophages and evaluate the role of ROS.Methods:H.pylori strain SS1 was co-cultured with the THP-1-derived macrophages at a multiplicity of infection (MOI) of 1∶100 based on trial results with different MOIs (ratios of THP-1 cells to bacteria ranging from 1∶25 to 1∶200).The co-culture supernatants and THP-1 cells were collected at various time points (3 h,6 h,12 h,24 h) and cytokine production was quantitated using ELISA analysis.The generation of intracellular ROS was detected by FCM,and the mRNA transcript levels of NLRP3 and caspase-1 were measured by Real-time PCR.Western blot was employed to analyze the expression of active caspase-1 subunit ( p10).Then we observed the inhibitory effects of NAC and siRNA specific for NLRP3 on the ex-pression of NLRP3 inflammasome-related components and the secretion of cytokines induced by H.pylori.Results:We found that H.pylori SS1 induced IL-1βand IL-18 production in human acute monocytic leukemia cell line THP-1 in a time-and dose-dependent manner.We further showed that H.pylori could induce the mRNA expressions of NLRP3 and caspase-1 in THP-1 cells.Moreover, release of IL-1βand IL-18 from H.pylori-infected THP-1 cells was suppressed by the ROS scavenger NAC,which was an agent known to inhibit NLRP3 inflammasome formation.NAC administration also resulted in a significant decrease in the level of H.pylori-induced caspase-1 protein expression in THP-1 cells.Additionally,secretion of IL-1βand IL-18 in response to H.pylori infection was remarkably reduced by NLRP3-siRNA.Conclusion:The induction of IL-1βand IL-18 secretion by H.pylori strain SS1 in THP-1 cells could be mediated through activation of NLRP3 inflammasome via ROS signaling pathway, which may be involved in the host innate immune defence and the pathogenesis of the bacteria.

Objective:To analyze the changes in T lymphocyte subsets, B lymphocytes and NK cells in children with active tuberculosis (TB) and their clinical significance.Methods:T lymphocyte subsets, B lymphocytes and NK cells in peripheral blood samples of 106 patients with acute TB (TB group) and 106 healthy children (healthy control group) were detected by flow cytometry and compared between different groups.Results:The percentages of CD3 + T, CD4 + T and NK cells as well as the CD4 +/CD8 + T cell ratio were significantly lower in the TB group than in the healthy control group ( Z=-3.783, P=0.000; Z=-5.401, P=0.000; Z=-3.434, P=0.001; Z=-2.014, P=0.044). The percentages of double negative T (DNT) and B cells in the TB group were significantly higher than those in the healthy control group ( Z=2.765, P=0.006; Z=6.880, P=0.000). No significant difference in the percentage of CD8 + T or double positive T (DPT) cells was observed between the two groups ( P>0.05). The expression of peripheral lymphocyte subsets varied in TB children of different age groups (0-<3, 3-<6, 6-<10 and 10-<16 years old). There were significant differences in CD3 + T, DNT and B cells among the four age groups ( H=10.081, P=0.018; H=14.583, P=0.002; H=8.498, P=0.037). The percentage of CD4 + T cells was significantly lower in children with extrapulmonary TB than in those with pulmonary TB ( Z=-3.068, P=0.002). No statistically significant difference in other lymphocyte subsets was found between children with extrapulmonary and pulmonary TB ( P>0.05). Conclusions:Tuberculosis could lead to immune dysfunction in children. Dynamic monitoring of the changes in peripheral lymphocyte subsets in children with TB could be conducive to better assessment of immune status and providing personalized treatment.

Objective To investigate the role of endometrial and subendometrial blood flows measured by transvaginal three-dimensional(3D) power Doppler ultrasound in the prediction of endometrial receptivity during in vitro fertilization-embryo transfer (IVF-ET) cycles. Methods One hundred and twenty patients undergoing IVF-ET cycles were enrolled in this study. Transvaginal 3D power Doppler ultrasound examination was performed on the day of human chorionic gonadotropin injection to determine endometrial thickness,endometrial volume, vascularization index(VI), flow index(FI) and vascularization flow index (VFI) of endometrial and subendometrial regions by experienced sonograpbers. The patients were divided into pregnant group and non-pregnant group according to pregnant outcome. The data between the two groups were compared and analysed. Results There were no significant differences( P ＞0.05) between the pregnant group and non-pregnant group in endometrial thickness, endometrial volume. Endometrial and subendometrial VI,FI, VFI were higher in the pregnant group than those in the non-pregnant group, and statistical comparison had significant difference( P ＜0.05). Conclusions Endometrial and subendometrial blood flows measured by transvaginal 3D power Doppler ultrasound can be as a reliable indicator to assess endometrial receptivity and to predict the pregnancy outcome.

Objective To understand the epidemic status quo and influencing factors of Kashin-Beck disease among children in mountain areas of Jilin Province.Methods Two hundred eighty-two severe endemic areas in 18 counties were selected and stratified by random cluster sampling method,and the status quo of KashinBeck disease prevalence was investigated among 7-12 year-old children according to the Diagnostic Criteria of Kashin-Beck Disease (WS/T 207-2010).In the meantime,the annual household income and the proportion of economic crops replanted,grain out-sourced,and returning farmlands to forests and grass were surveyed in the disease affected areas.Results A total of 14 162 children were investigated who had no clinical symptoms.Among them,28 cases were detected positive using X-ray with a detection rate of 1.98‰,while most of the cases were metaphysis positive.The annual household income (≥5 000 Yuan vs.＜ 5 000 Yuan) in the year 2009-2011 had a significant impact on the incidence of Kashin-Beck disease (1.47‰ vs.3.67‰,x2 =6.179,P ＜ 0.05),while the areas of returning farmland to forests and grass which accounted ＞ 1％ had no significant influence on the incidence compared with that ≤ 1％ (3.30‰ vs.1.57‰,x2 =3.876,P ＞ 0.05);the areas of economic crops replanting which accounted ＞ 10％ had no significant influence on the incidence compared with that ≤ 10％ (3.07‰ vs.1.65‰,x2 =2.565,P ＞ 0.05);the proportion of grain out-sourcing which accounted ＞ 50％ had no significant influence on the incidence compared with that ≤50％ (3.07‰ vs.1.65‰,x2 =2.565,P ＞ 0.05).Conehision Up to 2012,the disease among 7-12 year-old children of the mountain areas of Jilin Province have basically met the standard of Kashin-Beck disease elimination and the situation remains stable;furthermore,the household income has a significant impact on the incidence of Kashin-Beck disease.

This study was aimed to apply the electronic nose (E-nose) in the research of traditional Chinese medicine (TCM). The discussion was made on difficulties of using E-nose. The solution plan was proposed and the discrimination model was established. It provided a simple, rapid and effective analysi method in the identification of TCM. It also provided new ideas for the research and application of gas sensor arrays. E-nose was used in the ex-traction of TCM scent characteristics. Based on ion mobility spectrometry of MOS sensor, the fingerprint of TCM scent was established. The maximum response value of the sensor was used as analysis index. According to the diffi-culties of identification, two solution plans were proposed. Firstly, different detectors were employed to complete the classification. Secondly, radial basis function (RBF) and random forests (RF) were combined and then a cascade classifier was constructed in order to achieve the maximum of information obtained in conditions where the number of measurements, metal oxide semiconductor sensors in E-nose was limited. The results showed that both plans were accurate and practical with relatively high upper correct judge rate and better cross-validation (The highest upper correct judge rates were 95% and 100%, 96% and 80%, respectively). It was concluded that this study firstly ap-plied cascade classifier in the establishment of TCM identification by E-nose. With limited amount of sensors, the maximum information was received through data mining. Using E-nose in the identification of TCM was rapid and accurate. The established pattern recognition method was maneuverable with accurate identification rate and stability compared to conventional sensory identification method. It provided a simple and rapid analysis method for the iden-tification of TCM.