Objective To construct a luciferase reporter plasmid containing interferon regulatory factor 3(IRF-3)human gene promoter and to evaluate promoter activity in human embryonic kidney(HEK)-293 cells.Methods The 1 000 bp fragment was amplified by PCR with human genomic DNA as a template and was directionally cloned into pGL3-basic multiple cloning sites to construct the luciferase repor-ter plasmid pGL3-pIRF-3.Transfection of HEK-293 cells with the promoter-driven lucife-rase construct was performed to induce lucife-rase gene expression and calculate the relative luciferase activity unit(RLU).Promoter sequence of 1 000 bp upstream of transcription initiation site of IRF-3 was analyzed by using Promoter 2.0 Prediction software.Results DNA sequencing and restriction endonuclease analysis verified the successful construction of the plasmid pGL3-pIRF-3.This IRF-3 promoter exhibited a strong promoter activity with an increase of 42.2-fold of RLU in HEK-293 cells when compared with pGL-3 basic vector.The transfection experiment confirmed that the levels of its activation were significantly higher than that in controls in HEK-293 cells.Function analysis of IRF-3 promoter disclosed seve-ral GATA-1 and specific protein 1(Sp1) sites and E2F in minimal promoter region.Conclusion The plasmid pGL3-pIRF-3 promoter is successfully constructed and has a strong basal promoter activity in HEK-293 cells.

Objective: This study aims to investigate the correlation between the expression of FoxP3, TGF-β1, and epitheli-al-mesenchymal transition (EMT) in breast cancer and to determine the clinical significance of FoxP3. Methods: The expression of FoxP3, TGF-β1, E-Cadherin, N-Cadherin, Vimentin, and Fibronectin protein were detected in the cancer cells of 74 cases with breast carcinoma via immunohistochemistry. The correlation of FoxP3 protein with clinico-pathologic features of breast carcinoma and the re-lationships among the expressions of FoxP3, TGF-β1, and epithelial-mesenchymal transition (EMT) were analyzed. Results:The ex-pression rates of FoxP3, TGF-β1, and EMT are 36.5%(27/74), 39.2%(29/74), and 40.5%(30/74), respectively. The FoxP3 protein ex-pression in breast cancer is correlated with lymph node metastasis (P<0.05) but not with other clinico-pathological features (P>0.05). The expression of FoxP3 is also correlated with the expression of TGF-β1. Furthermore, TGF-β1 can induce EMT (P<0.05). Conclu-sion:The expression of FoxP3 is correlated with lymph node metastasis and EMT in breast cancer. Therefore, FoxP3 may be a marker for predicting metastasis.

One hundred and sixteen patients with lower limb fracture were screened by Doppler ultrasound for deep vein thrombosis(DVT) of bilateral lower extremities within the first 72 h, d7 and d21 after fracture. Results showed that DVT was detected in 31 (26. 7% ) out of 116 cases within 72 h; at d7 and d21 DVT was detected in 3 and 1 patient respectively with a cumulated DVT rate of 30. 2% in 3 weeks.Serial Doppler ultrasonography is of value in screening for DVT of the lower extremities in patients with lower limb fracture at early stage.

Objective:To observe the change of BDNF level expression in experimental in hippocampal and habenular nucles in experimental depression rats.Methods:The depression model of rats was established by chronic unpredictable mild stress. Openfield test was performed to detect the behavior of rats. Immunohistochemistry was used to observe the changes of BDNF level.Results:Compared to the control group,the number of depressed animals was much less than that of normal animals.Conclusion:The level of BDNF decrease significantly in experimental depressive model of rats.

OBJECTIVETo analyze the data of angle variation on traction based on a finite element model of complete cervical spine with straight physiological curvature, and try to give experimental reference and suggestion in treating cervical spondylosis.

METHODSA 43-year-old female patient with straight cervical spine was chosen and the CT scan data were collected. By using specially designed modeling system, a high quality finite element model of complete cervical spine with straight physiological curvature is generated,which included ligament and muscle according to anatomy. After the model was confirmed, traction was loaded with angle 0 degree, anterior 5 degrees, 10 degrees, 15 degrees, 20 degrees, 25 degrees, to observe the data of distance change on between adjacent intervertebral foramen, processus articularis, uncovertedral joint, intervertebral discs, and stress of anulus fibrosus and nucleus pulposus.

RESULTSWhen the angle was 0 degrees-15 degrees, the distance between intervertebral foramen, Luschka joint and processus articularis posterioris was enlarged, the tensile stress was adequate and compressive stress was small. It met the clinical requests.

CONCLUSION0 degree-15 degrees anterior position is suggested for the treatment of cervical spondylosis.

Adult ; Biomechanical Phenomena ; Cervical Vertebrae ; anatomy & histology ; surgery ; Female ; Finite Element Analysis ; Humans ; Spondylosis ; surgery ; Traction ; methods

OBJECTIVETo explore the effect of combination of Epimedii Folium and Ligustri Lucidi Fructus on osteoporosis rats induced by retinoic acid.

METHODSixty three-month-old male Wistar rats were randomly divided into the normal control group, the model group, the Epimedii Folium group, the Ligustri Lucidi Fructus group, the combination group of Epimedii Folium and Ligustri Lucidi Fructus and the raloxifene group. The osteoporosis model was established through oral administration with retinoic acid for two weeks. Meanwhile, all of treatment groups were administered with corresponding drugs for three weeks. The contents of serum calcium (Ca), phosphorus (P), alkaline phosphatase (AKP) and tartrate-resistant acid phosphatase (StrACP) were detected, and the pathomorphological changes of femurs were observed.

RESULTThe model control group showed much lower contents of serum Ca and P than the normal control group, but with significantly higher AKP and StrACP activity than the normal control group. The femoral head area showed reduced, narrow and sparse trabecular bones, with typical osteoporosis-like changes. Compared with the model control group, all of treated groups showed significant increase in Ca and P contents in serum, and down-regulate AKP and StrACP levels, while trabecular bones became more and wider, and densely interweaved as a reticular formation. Among them, the combination group showed the most significant effect.

CONCLUSIONEpimedii Folium and Ligustri Lucidi Fructus could effectively correct the abnormal bone metabolism and improve pathological conditions of bone tissues, so as to show the anti-osteoporosis effect. The combined application of the two drugs showed a better efficacy.

Acid Phosphatase ; blood ; Alkaline Phosphatase ; blood ; Animals ; Body Weight ; drug effects ; Calcium ; blood ; Drug Interactions ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Epimedium ; chemistry ; Femur ; drug effects ; pathology ; Isoenzymes ; blood ; Ligustrum ; chemistry ; Male ; Osteoporosis ; blood ; chemically induced ; drug therapy ; pathology ; Phosphorus ; blood ; Rats ; Rats, Wistar ; Tartrate-Resistant Acid Phosphatase ; Tretinoin ; adverse effects

AIM:To evaluate the application and effect of bandage contact lens in pterygium excision combined with conjunctival transplantation (CAT).METHODS:In a prospective, randomized, controlled clinical study, 110 patients (110 eyes) diagnosed with primary pterygium were collected in PLA NO.474 Hospital from January 2015 to January 2016.The 110 patients enrolled in the study in turn, and divided into two groups by the odd and even number.The odd number divided into bandage contact lens group (CAT + bandage contact lens, n=55), while the even number divided into control group (CAT, n=55).Visual analog scale (VAS) and corneal irritation were evaluated on 1, 3 and 7d after operation.Cornea fluorescent staining testing was carried out on 3d after operation.Following-up all the patients with 1a at least observed the recurrence of pterygium.RESULTS:The score of VAS in bandage contact lens group less than that in control group on 1d (4.13±2.06 vs 5.80±1.93, t=4.391, P<0.001) and 3d (2.09±1.36 vs 3.65±1.65, t=5.422, P<0.001) after operation, while there was no significant difference between two groups on 7d (t=1.295, P=0.198) after operation.The corneal irritation in bandage contact lens group less than that in control group on 1d and 3d after operation (P<0.05), while there was no significant difference between two groups on 7d after operation (P=0.052).Cornea fluorescent staining testing area in bandage contact lens group was less than that in control group on 7d after operation (0.33±0.37mm2 vs 2.73±2.21mm2, t=7.921, P<0.01).There was no significant difference in recurrence rate between two groups after 1a operation (P=1.000).CONCLUSION:Bandage contact lens could significantly release pain and corneal irritation, promote the healing of the corneal epitheliums in the defected area, and increase the postoperative comfort level in patients after operation.

The study was using the orthogonal test and making the extraction rates of icariin, ferulic acid, epimedin A, epimedin B, epimedin C, baohuoside I and ligustilide determinated by HPLC multiwavelength switch, gradient elution and multi-index comprehensive weighted scoring method (weight coefficient was 0.47: 0.16: 0.07: 0.07: 0.08: 0.06: 0.09) as evaluation index, combine with SPSS 16.0 software to optimizing the best extraction. It was Yinpian soak 1 h, 12 times more than the volumn of 50% ethanol solution, by heating reflux extraction for 60 min. The compliance test indicates that the optimized compatibility extraction technology is stable and practical, and it has provided an experimental basis for compound preparation technology research of Epimedium brevicornu and Ligusticum chuanxiong.
Chemical Fractionation ; methods ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Epimedium ; chemistry ; Ligusticum ; chemistry

ABSTRACT:Objective To investigate the effect and mechanism of Hedgehog signaling pathway on the invasion of breast cancer cells in vitro.Methods The SHH,SMO and Gli-1 expression levels of breast cancer cell line MDA-231 and normal mammary epithelial cell line MCF-10A were detected by Western blot at protein level and by Real-time RT-PCR at mRNA level.Next,shRNA vector was transfected into the MDA-2 3 1 cells with highly expressed SMO,and the stable transfected cells were selected by G4 1 8 .Western blot and Real-time RT-PCR were performed to observe the inhibitory effect of RNAi on SMO expression.MTT assay was used to assess the influence of SMO siRNA on cell proliferation.Transwell assay was applied to observe cell invasion ability.The expressions of Gli-1,Snail,MMP-9,E-cadherin and Vimentin protein were determined by Western blot.Results Compared with those of normal mammary epithelial cell line MCF-10A,the expressions of SHH,SMO and Gli-1 were significantly increased.The invasion of MDA-2 3 1 cells was inhibited significantly after SMO silencing.Additionally, the protein expressions of Gli-1 , Snail, MMP-9 and Vimentin were obviously inhibited, and E-cadherin was significantly increased.Conclusion Mutative activation of Hedgehog signaling pathway in breast cancer cells promotes cell invasion probably through induction of epithelial-mesenchymal transition of the tumor cells.

Objective To observe the improvement effects of Qingshen Granule on the life quality of chronic kidney disease (CKD) patients with damp-heat syndrome. Methods The CKD patients with damp-heat syndrome were randomly divided into experimental group and control group. The control group was treated with Western medicine, and the treatment group was treated with Qingshen Granules additionally for 12 weeks. SCr and eGFR were observed before and after treatment, and the life quality was investigated by KDQOL-SFTM1.3. Results Actually 156 cases were completed, including 77 cases in the treatment group and 79 cases in the control group. The total rate of curative effect was 81.82% (63/77) in teatment group, and (63.29%, 50/79) in control group, with significent difference (P<0.05). Multivariate linear regression analysis indicated that gender, education level, TCM syndrome score, CKD stage, eGFR and Hb level could affect the life quality. The scores of KDQOL-SFTM and its containing the MOS item short from health survey (SF-36) and kidney disease targeted areas (KDTA) were significantly improved after treatment in experimental group (P<0.05), while the scores in control group were with no significant improvement. The improving effects of PCS, and SPL, SLEEP, PS in KDTA in treatment group were significantly superior to those in control group (P<0.05). The scores of life quality in effective and stable cases increased significantly after treatment compared with before treatment (P<0.05). Conclusion Qingshen Granules can effectively improve the life quality of CKD patients with damp-heat syndrome, and the effect is not dependent on the improvement of laboratory indexes (renal function).