Aim ToinvestigatetheeffectsofmiRNA-181 a on breast cancer resistance protein ( BCRP ) . Methods Bioinformaticspredictedbindingsitesof BCRP mRNA-3ˊUTR region and miR-181 a;further lu-ciferase reporter gene analysis confirmed that miR-181 a could combine with BCRP mRNA-3ˊUTR; qRT-PCR and Western blot detected related mRNA and protein expressionlevels.Results Comparedwithnegative transfection group, after the miR-181a mimic and PGL3-BCRP 3ˊUTR were co-transfected, luciferase ac-tivity was significantly decreased ( P <0 . 05 ) . After miR-181a mimic transfected MCF7/MX cells for 48h, compared to the negative group, the expression of miR-181 a in MCF-7/MX cells was increased ( P<0. 05 ) , BCRP mRNA, BCRP protein expression were signifi-cantly decreased ( P<0 . 05 ) , while mRNA expression and protein levels of MRP, P-gp, LRP did not change significantly ( P >0. 05 ); after transfecting miR-181 a inhibitor for 48h, compared to the negative group, the expression of miR-181 a in MCF-7 cells was reduced (P<0. 05). Meanwhile,BCRP mRNA expression and BCRP protein expression were also significantly in-creased ( P<0. 05 ) . The mRNA expression and pro-tein levels of MRP, P-gp, LRP did not change signifi-cantly(P>0.05).Conclusion miR-181acanregu-late BCRP expression by targeting the BCRP mRNA-3ˊUTR region.

Objective To quantitatively evaluate the left ventricular systolic synchrony in patients with chronic renal failure (CRF) by real-time three-dimensional echocardiography (RT-3DE). Methods Thirty patients with CRF and twenty-five normal subjects were enrolled in this study.The M-mode ejection fraction (M-EF), left ventricular end diastolic internal diameter (LVIDd), interventricular septum diameter (IVSd), left ventricular posterior wall diameter (LVPWd) were obtained on two-dimension. The global and regional volume-time curves were obtained on three -dimensional. The end diastolic volume (LVEDV),end systolic volume (LVESV) and ejection fraction (LVEF) of left ventricule, the time to minimal systolic volume (Tmsv) of 16,12,6-segmental standard deviation (Tmsv16, 12, 6-SD), maximal difference (Tmsv16, 12, 6-Dif) were derived from Qlab software . The above parameters as a percentage of the cardiac cycle with different heart rates between patients were also calculated from the Qlab software, which were Tmsv16, 12, 6-SD% and Tmsv16, 12, 6-Dif %, rspectively. Results The indices of HR, LVIDd, IVSd, LVPWd and LVESV were significantly higher while LVEF was significantly lower in the CRF group than those of the control group (P<0.05 or P<0.01); The indices of systolic asynchrony of Tmsv16,12-SD%, Tmsv16,12-Dif, Tmsv16 ,12 ,6-SD/R-R%,Tmsv16,12,6-Dif/R-R% were significantly larger in the CRF group than those of the control group(P<0.05 or P<0.01). Conclusions RT-3DE provides a simple, intuitional and noninvasive approach to assess the systolic synchrony of all the LV segments simultaneously and LVEF in patients with chronic renal failure.

AIM: To evaluate the changes of corneal high - order aberration (including Coma, Spab, RMSh) after laser in situ keratomileusis (LASIK) with femtosecond laser, sub- Bowman keratomileusis ( SBK ) and laser epithelial keratomileusis (LASEK). METHODS: Of 82 myopic patients ( 164 eyes ), 31 patients (62 eyes) were treated by FS-LASIK, 31 patients (62 eyes) were treated by SBK, 20 patients (40 eyes) were treated by LASEK. Sirius system was used for measuring the coma aberration, spherical aberration, and high order aberration at 1, 15d,1, 3mo after surgery. RESULTS: 1) Vision: The uncorrected visual acuity of the three groups had no differences (P>0. 05). 2) Corneal aberrations: Three kinds of surgical procedure for patients with corneal aberration had significant impact. The C7, C8, C12 and RMSh of three groups were increased significantly (P<0. 05). The C7, C8, C12 and RMSh were not recovered to preoperative levels after 3mo. But the increase of patients after FS- LASIK was smaller than the other two groups, with statistical significance (P<0. 05). CONCLUSION: Compared with SBK and LASEK, FS - LASIK has better visual acuity in the early postoperative and corneal higher-order aberrations increase is relatively small.

[Summary] Graves'ophthalmopathy ( GO) is the most common extrathyroidal manifestation of Graves disease ( GD) . It is an organ-specific autoimmune disorder. The estimated incidence of GO in the general population is 16 women and 3 men per 100,000 population every year. Approximately 3%-5% of patients with GO are exposed to the potential of losing their sight and requiring optimized medical intervention. For the clinicians it is helpful to assess the degree of activity and severity of GO;both are important in deciding whether a patient requires intervention and which type of intervention is indicated. These interventions include management of hyperthyroidism, measures to relieve local symptoms, immunosuppressive therapies for active moderate and severe GO, and finally rehabilitative surgery for inactive patients.

OBJECTIVETo investigate the accumulation of advanced glycation end products (AGE) and the inflammatory response of skin and wound in diabetic patients, and to analyze their relationship in vitro.

METHODSHistological staining and immunohistochemical staining was respectively performed on skin and wound tissue specimens collected from 10 patients with Type II diabetes mellitus (diabetes group) and 12 non-diabetic patients with skin injury (control group) to observe the arrangement of collagen and the distribution of inflammatory cells, and to determine the expression levels of AGE and its receptor (RAGE). Malondialdehyde (MDA) levels in skin and wound tissue homogenates were assayed by enzyme-linked immunosorbent assay. In vitro, human neutrophils were isolated and treated with RPMI-1640 culture medium or that containing AGE-human serum albumin in the concentration of 0.315, 0.625, 1.250 mg/mL, and they were identified as normal control (NC) group, low concentration (L) group, moderate concentration (M) group, and high concentration (H) group. Cell viability in each group was determined by MTT colorimetric assay, and the reactive oxygen species (ROS) in cell was measured with 2', 7'-dichlorofluorescein-diacetate. Data were processed with t test.

RESULTSCompared with those of skin in control group, collagens of skin tissues in diabetes group atrophied and disorderly arranged. Inflammatory cells in wounds in diabetes group were dispersed, in which collagens arranged loosely and irregularly, as compared with those of wounds in control group. Expression levels of AGE and RAGE of skin in diabetes group were higher than those in control group. In diabetes and control groups, especially in diabetes group, the numbers of RAGE-positive cells in wound tissue were more than those in skin tissue. Large amount of inflammatory cells with positive expression of RAGE were observed in diabetes group. MDA level of skin and wound tissue in diabetes group was respectively (6.3 ± 1.0), (7.1 ± 2.4) nmol per milligram protein, which were obviously higher than those in control group [(2.9 ± 1.0), (3.6 ± 1.4) nmol per milligram protein, with t value respectively 8.017, 4.349, P < 0.05 or P < 0.01]. Cell viability and ROS levels in neutrophils were increased in L, M, and H groups [(59 ± 8)%, (77 ± 5)%, (67 ± 6)% and 1.67 ± 0.14, 2.13 ± 0.17, 3.48 ± 0.48] as compared with those in NC group [(34 ± 5)% and 0.58 ± 0.06, with t value respectively 7.195, 14.890, 11.130 and 20.195, 24.905, 16.864, P < 0.05 or P < 0.01].

CONCLUSIONSAbnormal oxidative stress in diabetic skin leads to an atypical origin of wound repair. AGE-RAGE effect is a critical mediator for oxidative stress in diabetic wound tissue during wound healing.

Aged ; Case-Control Studies ; Diabetes Mellitus, Type 2 ; metabolism ; pathology ; Female ; Glycation End Products, Advanced ; metabolism ; Humans ; Male ; Middle Aged ; Oxidative Stress ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Serum Albumin ; metabolism ; Serum Albumin, Human ; Skin ; metabolism ; pathology ; Wound Healing

The combination of Danggui and Honghua (GH) is a popular herb pair commonly used in clinic for the treatment of blood stasis syndrome in China. To evaluate the activating blood circulation and dissipating blood stasis effects of the combination of different proportions of Danggui and Honghua on acute blood stasis rats, and optimize the proportion of GH to have the best activating blood circulation and dissipating blood stasis effect. Acute blood stasis rat model was induced by subcutaneous injection of adrenaline and ice water bath. The blood stasis rats were administrated intragastrically with GH (1 : 0, 4 : 1, 2 : 1, 3 : 2, 1 : 1, 2 : 3, 1: 2, 1 : 4 and 0 : 1) extracts. The whole blood viscosity (WBV), plasma viscosity (PV), and high shear whole blood relative index (HSWBRI), low shear whole blood relative index (LSWBRI), and erythrocyte aggregation index (EAI) were tested to observe the effects of GH on hemorheology of blood stasis rats. And the maximum aggregation induced by adenosine diphosphate (ADP) was tested to observe the effect of GH on platelet aggregation index of blood stasis rats. In addition, the prothrombin time (PT), thrombin time (TT), activated partial thromboplastin time (APTT) and plasma fibrinogen (FIB) were tested to observe the effects of GH on blood coagulation function of blood stasis rats. Then principal component analysis and multi-attribute comprehensive index methods were both used to comprehensively evaluate the total activating blood circulation and dissipating blood stasis effects of GH. The results showed that the hemorheological indexes and coagulation parameters of model group both had significant differences with normal group. Compared with model group, GH (1 : 0, 4 : 1, 2: 1, 3 : 2, 1 : 1, 2 : 3, 1 : 2, 1 : 4 and 0 : 1) could improve all the blood hemorheology indexes and regulate part indexes of blood coagulation function and platelet aggregation in acute blood stasis rats. Based on principal component analysis and multi-attribute comprehensive index methods, GH 1 : 1 and GH 3 : 2 both had the best effect of blood circulation and dissipating blood stasis, and the effect of GH 1 : 1 was slightly better than GH 3 : 2. These results suggest that GH could obviously ameliorate the abnormality of hemorheology and blood coagulation function in acute blood stasis rats. The optimized proportion of GH was consistent with regulations of medicine usage that GH 1 : 1 had the highest frequency used in traditional Chinese formulae. It could provide scientific basis for more effective application of the compatibility between Danggui and Honghua in modern clinic medicine.
Animals ; Blood Coagulation ; Blood Viscosity ; Carthamus tinctorius ; China ; Drugs, Chinese Herbal ; pharmacology ; Erythrocyte Aggregation ; Hemorheology ; Partial Thromboplastin Time ; Platelet Aggregation ; Principal Component Analysis ; Prothrombin Time ; Rats ; Rats, Sprague-Dawley ; Thrombin Time

The different biological functions were studied in mouse bone marrow-derived endothelial progenitor cells isolated by differential time attachment to obtain the optimal adherent time in this study. Density gradient centrifugation-isolated bone marrow mononuclear cells were seeded on the fibronectin-coated dish. The 1-day cultured unattached cells were seeded on the second dish for 2 more days. Then unattached cells in the second dish were seeded on the third dish. The cells on 3 dishes were defined as 1-day adherent cells, 3-day adherent cells and 3-day unattached cells, respectively. After 20-day culture, the biological functions, such as the percentage of biomarkers, the ability of adhesion, and the ability of forming tubes in vitro were analyzed. The results showed that the percentages of positive CD34, FLK-1, and CD34/FLK-1 expressions in 1-day attached cells were significantly increased compared to those in the 3-day adherent or unattached cells (P < 0.01), which showed the strongest adhesion ability. The expression of eNOS in 1- or 3-day adherent cells was significantly higher than that in 3-day unattached cells (P < 0.01). The expression of VEGF in 3-day adherent cells was significantly higher than that in 1-day adherent cells or 3-day unattached cells (P < 0.01). These results suggest the biological functions of 1-day adherent cells are significantly stronger than that of 3-day adherent or unattached cells. VEGF expression in 3-day adherent cells is higher than that in 1-day adherent cells or 3-day unattached cells. The expression of eNOS in 1-day adherent cells or 3-day adherent cells is higher than that in 3-day unattached cells. The optimal adherent time to obtain mouse bone marrow-derived endothelial progenitor cells is 1-3 d.
Animals ; Bone Marrow Cells ; cytology ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Separation ; methods ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Leukocytes, Mononuclear ; cytology ; Male ; Mice ; Mice, Inbred C57BL ; Nitric Oxide Synthase Type III ; metabolism ; Stem Cells ; cytology ; metabolism ; Time Factors ; Vascular Endothelial Growth Factor A ; metabolism

Sciatic nerve injury is a common disease of peripheral nerve in clinic. After nerve injury, there are many dysfunctions in motoneurons and muscles following regeneration. Previous studies mostly investigated the aspects related to the injured nerve, and the effect on the recurrent inhibition (RI) pathway of spine following regeneration was not fully understood. Following reinnervation after temporary sciatic nerve crush, the functional alteration of RI was studied. In adult rats, RI between lateral gastrocnemius-soleus (LG-S) and medial gastrocnemius (MG) motor pools was assessed by conditioning monosynaptic reflexes (MSRs) elicited from the cut dorsal roots and recorded from either the LG-S or MG nerves by antidromic stimulation of the synergist muscle nerve. The following results were obtained. (1) The RI of MSRs in rats was almost lost (<5 weeks) after sciatic nerve crush. Although the RI partially recovered following reinnervation (6 weeks), it remained permanently depressed (up to 14 weeks). (2) Sciatic nerve crush on one side did not affect the contralateral RI. (3) Sciatic nerve crush did not induce any motoneuron loss revealed by immunohistochemistry. Peripheral nerve temporary disconnection causes long term alterations in RI pathway which make up motoneuron's function enhance for the alteration of muscle power and suggests that peripheral nerve injury induces long term plastic changes in the spinal motoneuron circuitry.
Animals ; Long-Term Synaptic Depression ; physiology ; Male ; Motor Neurons ; physiology ; Nerve Crush ; Nerve Regeneration ; physiology ; Neuronal Plasticity ; physiology ; Neurons, Afferent ; physiology ; Rats ; Rats, Wistar ; Reflex, Monosynaptic ; physiology ; Sciatic Nerve ; injuries ; physiopathology ; Spinal Cord ; physiopathology ; Spinal Nerve Roots ; physiopathology

OBJECTIVETo investigate the repeatability of three-dimensional (3-D) cephalometric measurements for the clinical application of 3-D cephalometry.

METHODSForty-nine measurements that widely used in traditional cephalometric analyses were defined in 3-D cone-beam CT (CBCT) images. Three examiners identified landmarks on CBCT images of 17 subjects with normal occlusion, respectively, and 3-D measurements were exported automatically by software SimPlant. Inter-examiner reliability correlation coefficients (ICC) were obtained for all measurements.

RESULTSRepeatability of 36 measurements was high (ICC value greater than 0.9), including SNA, SNB. Repeatability of 11 measurements was moderate (ICC value between 0.8 and 0.9), including CoL-GoL, CoL-MSP. Repeatability of 2 measurements was low (ICC value lower than 0.8), including Gn-MSP and MPR-MSP.

CONCLUSIONSMost 3-D cephalometric measurements based on CBCT had high repeatability. However, some 3-D cephalometric measurements had limited repeatability.

Cephalometry ; Cone-Beam Computed Tomography ; Humans ; Imaging, Three-Dimensional ; Observer Variation ; Reproducibility of Results

OBJECTIVETo explore the time law of electroacupuncture in regulation of circadian rhythms of the organism.

METHODSEffects of electroacupuncture at "Shenshu" (BL 23) at Zi, Wu, Mao and You periods on circadian rhythms of locomotor activity and core body temperature in hamsters were observed with chronobiological research methods.

RESULTSElectroacupuncture at Wu period could decrease the amplitude of locomotor activity rhythm (P < 0.05), at Mao period could delay the peak phase of circadian rhythm and at You period could advance the peak phase of circadian rhythm (both P < 0.05); and electroacupuncture at Mao period could delay 22.36 degrees and at You period advance 39.32 degrees for the rhythm peak of the circadian rhythm of core body temperature.

CONCLUSIONAcupuncture has a certain effect on circadian rhythm of locomotor activity and core body temperature.

Acupuncture Therapy ; Animals ; Body Temperature ; Body Temperature Regulation ; Circadian Rhythm ; Cricetinae ; Motor Activity