Aim To investigate the effect of Rhein derivative 4a containing amide structure on migration and invasion in ovarian cancer SKOV3 cells and its possible mechanism. Methods Ovarian cancer SK0V3 cells were used as target cells. Molecular docking and West-em blot were used to detect the regulatory effect of derivative 4a on Racl protein. CCK8, HE staining, Scratch and Transwell assay were used to detect the effects of derivative 4a on the proliferation, morphology , migration and invasion of SK0V3 cells, respectively. Western blot was employed to determine the expression of matrix metalloproteinases and EMT-related proteins. Results Derivative 4a could effectively bind to Racl protein, and the binding energy was-29. 10 kcal • mol"1, which was significantly lower than that of Rhein; it also could down-regulate the expression of Racl protein in SK0V3 cells. Derivative 4a could significantly inhibit the proliferation, invasion and migra tion of SKOV3 cells, and induce a large amount of cellular vacuolation; derivative 4a could also down-regu-\ late the expression of MMP-2 and MMP-9, up-regulate the expression of EMT epithelial marker protein E-ca-derin but down-regulate the expression of vimentin and j3-cantenin. Conclusions Derivative 4 a can inhibit the proliferation, migration and invasion of ovarian cancer SK0V3 cells. The mechanism may relate to its targeted regulation of Racl, thereby inhibiting the secretion of matrix metalloproteinases, up-regulating the expression of key molecule E-caderin and down-regula-ting the expression of Vimentin and (3-cantenin in EMT i process.•.

Objective To investigate the DNA methylation of skeletal muscle in spastic paralysis rats and correlation with the muscle fiber configuration. Methods 100 5-day old Wistar rats were randomly divided into model group and control group. The former was established the spastic paralysis modle and reared for 30 days. Then, tissues from the gastrocnemius of all the rats were observed with triplicate DNA methylation, myosin heavy chain-I (MHC-I) mRNA with RT-PCR and transmission electron microscopy. Results The DNA methylation was (4.95±0.83)×10% in the model group, significantly less than (6.59±0.75)×10% in the control group (P<0.001); while the MHC-I mRNA was (1.23±0.31), significantly more than (0.44±0.29) in the control group (P<0.001). The Z-line was disordered, and the mitochondria near the Z-line increased, with edema and partially broken in cristae. The balance between the thick and thin filaments was broken, and myofibrils envelope fused. Conclusion Hypomethylation and hyperexpression of MHC-I mRNA have been found in skeletal muscle of spastic paralysis rats, which may result in type I fibers increase. However, there was no sufficient evidence to support the correlation between the DNA methylation and the secondary pathological changes.

Objective To investigate the role of CD4~+CD25~+ Treg cells on the pathogenesis of ankylos- ing spondylitis(AS), and to study the machanism of tumor necrosis factor(TNF)-?blocker on the treatment of AS by detecting the number of CD4~+D25~+ Treg cells before and after the treatment. Methods The diagno- sis of 10 AS patients was made based on the 1984 modified New York criteria. The patients received subcuta- neou injection of recombinant human tumor necrosis factor receptor-Fc fusion protein(rhTNFR-Fc)(etaner- cept)50 mg weekly for 8 weeks and 10 heathy subjects were enrolled for control. The mononuclear cells were isolated from peripheral blood in beth patients and controls. The number of CD4~+CD25~+ T cells and CD4~+ CD25~(high)T cells and the expression of CTLA-4. were detected by flow cytometry. Results The proportion of CD4~+CD25~+ T cells(24?19)% in total CD4~+ T lymphocytes of peripheral blood and CD4~+CD25~(high)T/CD4~+ T (6?6)% from AS patients before treated with rhTNFR-Fc was higher than that in healthy volunteers and AS patients after treatment(P

Background Neovascular glaucoma (NVG) is a serious ocular disease which may cause blindness.The primary pathogenesis of NVG is ischemic retinopathy derived by central retinal vein occlusion (CRVO) and diabetic retinopathy (DR).Clinical characteristics of NVG are variable based on the difference of primary diseases,such as CRVO and DR.However,there is a few studies regarding the diffcrcnces of NVG initiated by CRVO and DR.Objective This study was to compare the clinical characteristics in NVG patients secondary to CRVO and DR.Methods A series case observational study was carried out in Hiserve Hospital of Qingdao University from January 2009 to June 2012.Twenty-nine eyes of 27 patients with NVG caused by CRVO (10 eyes of 10 patients) and DR (19 eyes of 17 patients) were included.The history of underlying diseases,course of NVG,intraocular pressure(IOP),fundus findings and complications after treatment were analyzed and compared between the CRVO-derived NVG and DR-derived NVG.All patients underwent panretinal photocoagulation,improving microcirculation therapy,anti-glaucoma (drug or surgery) and causative disease treatment,and some of them received vitrectomy or/and cataract surgery.Two eyes from each group received intravitreal injection of ranibizumab.The follow-up time in both groups was (14.00±10.13) months and (17.89±12.52) months,respectively.Results The median time of underlying disease was 3.3 months (2 weeks to 6 months) in the CRVO patients and 11.1 months (4 to 36 mouths) in the DR patients,with a significant difference between them (Z =-2.40,P＜0.05).CRVO-derived NVG progress was much faster than that of DR-derived NVG.The number of the eyes with visual acuity improvement after treatment was 2 in the CRVO-derived NVG and 15 in the DR-derived NVG;while the number of the eyes with unchanged or worse visual acuity was 8 and 4 in the CRVO-derived NVG eyes and the DR-derived NVG eyes (x2 =9.38,P＜0.01).The difference of IOP in pre-and post-treatment was (37.00±9.91)mmHg in the CRVOderived NVG eyes and (8.92±12.05)mmHg in the DR-derived NVG eyes,showing a significant difference between them (t =6.30,P＜0.01).In the CRVO-derived NVG eyes,optic disc edema,retinal hemorrhage,and vein dilatation were seen in 6 eyes,and mild optic disc edema and retinal hemorrhage were observed in 4 eyes.After treatment,fundus could not be seen in 4 eyes,in other 2 eyes optic disc and retinal laser spots were unclearly observed.In addition,pale optic disc and retinal vessel occlusion appeared in 2 eyes,and silver wire-like arteries exhibited in 2 eyes.In pre-treated DR-derived NVG eyes,fundus could not be seen in 8 eyes and Ⅲ-Ⅳv stages of DR findings appeared in 11 eyes.After treatment,retinopathy was stabilized in 16 eyes of 15 cases.Advanced retinopathy(V-Ⅵ stages of DR findings) was revealed in 3 eyes of 3 cases.The incidence of the complication after treatment was 100.0％ in the CRVO-derived NVG eyes and 21.1％ in the DR-derived NVG eyes (x2=5.18,P＜0.05).Conclusions The clinical characteristics of NVG secondary to CRVO and DR are variable,an appropriate treatment option should be selected according to different features of NVG.

OBJECTIVETo observe the differences of therapeutic effects on common cold of wind-cold type at early stage treated by different cupping duration.

METHODSSixty cases of common cold of wind-cold type were randomly divided into an observation group and a control group by fifty-fifty. Flash cupping was applied at Dazhui (GV 14), Fengmen (BL 12), Feishu (BL 13), Xinshu (BL 15) and Geshu (BL 16), etc., and the cupping was retained for 25-30 min in the observation group and 15 min in the control group. The therapeutic effects were compared after twice treatments.

RESULTSThe total effect rate was 93.3% (28/30) in observation group, superior to that 66.7% (20/30) in control group (P < 0.01).

CONCLUSIONThe therapeutic effect of common cold of wind-cold type at early stage treated with cupping for 25-30 min is obvious, superior to that for 15 min.

Acupuncture Points ; Adolescent ; Adult ; Common Cold ; therapy ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Treatment Outcome ; Young Adult

BACKGROUNDParkinson's disease (PD) is an autosomally inherited neurodegenerative disease in elderly people. The etiology of PD has long been thought to be associated with both genetic and environmental factors. To explore potential genetic risk factors for PD in the northern Han Chinese population, we investigated three single nucleotide polymorphisms (SNPs) (rs4538475, rs11107 and rs12564040) in the BST1, PARK15 and PARK9 genes.

METHODSGenomic DNA from 215 PD patients and 212 matched controls was amplified in two independent PCR systems and subsequently genotyped by digestion with the endonuclease PstI. Genetic parameter and association studies were carried out with SPSS 13.0 and PLINK 1.07 software.

RESULTSWe could accurately detect all genotypes in the three loci with the PCR-RFLP or mismatched PCR-RFLP techniques. The observed heterozygosities of the rs4538475 and rs11107 loci in PD and control groups ranged from 0.460 - 0.481 and 0.410 - 0.441, in BST1, PARK15 respectively, while we detected no heterozygosity at the rs12564040 locus in PARK9. The similar distributions of genotypic frequency between both groups suggest that the three SNPs investigated in this study are unlikely to play roles as common risk factors or pathogenic mutations for PD in northern Han Chinese.

CONCLUSIONThe SNPs investigated in the BST1, PARK15 and PARK9 genes associated with PD susceptibility are not associated with PD in the northern Han Chinese population.

ADP-ribosyl Cyclase ; genetics ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; genetics ; Asian Continental Ancestry Group ; genetics ; F-Box Proteins ; genetics ; Female ; GPI-Linked Proteins ; genetics ; Genetic Predisposition to Disease ; genetics ; Humans ; Male ; Middle Aged ; Parkinson Disease ; genetics ; Parkinsonian Disorders ; genetics ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; genetics ; Young Adult

OBJECTIVETo observe the clinical effect of Dihuang Zhixue Capsule (DZC, a Chinese preparation for cooling blood and dispelling toxic substances) in the treatment of childhood refractory idiopathic thrombocytopenic purpura (RITP), with cyclosporin A (CsA) used as the control.

METHODSForty-one children of RITP were randomized into the treated group and the control group. The 21 patients in the treated group were orally given 2 to 3 DZC capsules each time, thrice a day and the 20 in the control group were given 3 mg/kg CsA per day, with 3 months as one therapeutic course. The therapeutic efficacy, platelet count and adverse reaction in the two groups were compared at the end of the course.

RESULTS(1) In the treated group, 1 (4.8%) patient was evaluated as cured, 3 (14.3%) as markedly effective, 5 (23.8%) as effective, 5 (23.8%) as improved, 7 (33.3%) as ineffective, with the total effective rate being 66.7%; while in the control group, the corresponding numbers were 0, 2 (10.0%), 2 (10.0%), 3 (15.0%), 13 (65.0%) and 35.0%, respectively, showing statistical significance in difference between the total effective rates of the two groups (xi(2)=4.11, P=0.0426). (2) As compared with the baseline, the platelet count increased in both groups after 2 months' treatment (P<0.05). After 3 months' treatment, the platelet count was higher in the treated group than in the control group (P<0.05). (3) The improvement of hemorrhage in the treated group after 8 weeks' treatment was better than that in the control group (P<0.05). (4) No apparent adverse reaction was observed in the treated group, while in the control group, hirsutism was shown in 15 cases; gingival hyperplasia in 10; digestive reaction in 5, liver function impairment in 5, hypertension in 2 and renal impairment in 2.

CONCLUSIONThe therapeutic efficacy of DZC is better than that of CsA, and DZC shows good compliance but brings no obvious adverse reaction.

Adolescent ; Capsules ; Child ; Child, Preschool ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Hemorrhage ; drug therapy ; Humans ; Male ; Platelet Count ; Purpura, Thrombocytopenic, Idiopathic ; drug therapy ; Treatment Outcome

Objective:To study the expression of IL-21/BCL-6/Blimp-1 in CE patients and discusse the mechanism of the pathogenesis of the echinococcosis.Methods:27 patients and 30 health persons were collected from the first affiliated hospital of Xinjiang medical university in the same period.IL-21 was detected by ELISA and the expression of IL-21/BCL-6 /Blimp-1 mRNA was detected by Real-time fluorescence quantitative PCR (qRT-PCR) in CE patients.At the same time,17 patients were followed up in the group of patients,and the expression of IL-21/BCL-6/Blimp-1 was detected before and after treatment.Results:(1) The results of PCR showed that the levels of IL-21/BCL-6 mRNA were significantly increased in the peripheral blood mononuclear cells of the CE patients compared with the healthy control group (P<0.05).The expression of IL-21/BCL-6 /Blimp-1 mRNA in patients before the treatment was higher than that of patients after treatment(P<0.05).(2)The level of IL-21 in peripheral blood of CE patients was sig-nificantly higher than that in the healthy control group and basically returned to normal after the cure (P<0.05).IL-21 was positively correlated with BCL-6(r=0.733, P<0.01).Conclusion:BCL-6 and Blimp-1 May promote the human immune system to resist parasitic infection in the course of the development of the disease.IL-21, BCL-6 and Blimp-1 are significantly reduced after effective treatment,indicating that these factors are involved in the immune mechanism of the development of the disease.

Human bone marrow is the major source of mesenchymal stem cells (MSC). It was reported that the standard density gradient centrifugation method was not efficient in isolating MSC and it may be caused by the existing of bone marrow particles. In previous study, a lot of MSC were obtained by culturing bone marrow particles alone combined with standard method. However, it is time- and labor-consuming to obtain bone marrow particles by filtering and to isolate MNC by density gradient centrifugation. This study was purposed to explore the more simple and efficient method to isolate MSC from bone marrow. Seven normal bone marrow aspirates were collected and centrifugated. The bone marrow particles floated on surface layers were cultured by modified primary explant culture, whereas the bone marrow aspirates deposited were cultured by direct plating method, then the immun phenotype and differentiation capability of isolated cells were analyzed. The results showed that in 3 of 7 aspirates, bone marrow particles were floated on surface layers, whereas the other bone marrow cells and some particles were deposited after centrifugation. The MSC were reliably isolated from the floating layers or deposited aspirates by modified primary explant culture and direct plating method separately. After 3 passages the isolated MSC did not express CD45 and CD34, but expressed CD105, CD73, CD44, CD90, CD49e and they could differentiate into chondrocytes and adipocytes. It is concluded that normal human bone marrow MSC can be isolated simply and efficiently by direct plating method in combination with modified primary explant culture.
Bone Marrow Cells ; cytology ; Cell Culture Techniques ; methods ; Cell Separation ; methods ; Cells, Cultured ; Humans ; Mesenchymal Stromal Cells ; cytology

OBJECTIVETo observe the difference in the efficacy on lumbar intervertebral disc protrusion (LIDP) between Santong tuina therapy and conventional tuina therapy.

METHODSMulti-central, random and controlled clinical trial was carried out. One hundred and twenty cases of LIDP were randomized into an observation group and a control group, 60 cases in each one. In observation group, Santong tuina therapy was used. In control group, conventional tuina therapy was adopted. The clinical efficacy was observed in two groups, and lumbar vertebral function was scaled before and after treatment in two groups.

RESULTSThe total effective rate was 95.0% (57/60) in observation group and was 96.7% (58/60) in control group, without significant statistical difference in comparison (P > 0.05). The cured and markedly effective rate was 81.7% (49/60) in observation group and was 63.3% (38/60) in control group, indicating significant statistical difference in comparison (P < 0.05). The self-comparison of lumbar vertebral function before and after treatment presented statistical significance in either observation group (12.25 +/- 3.15, 25.56 +/- 5.27) or control group (13.32 +/- 3.26, 20.46 +/- 4.25, both P < 0.05); additionally, there was significant difference in the comparison between groups after treatment (P < 0.05). The therapy adopted in observation group improved lumbar vertebral function much significantly.

CONCLUSIONIn the treatment of LIDP, Santong tuina therapy achieves much better clinical efficacy as compared with conventional tuina therapy.

Adolescent ; Adult ; Aged ; Female ; Humans ; Intervertebral Disc Displacement ; therapy ; Lumbar Vertebrae ; Male ; Manipulation, Orthopedic ; methods ; Medicine, Chinese Traditional ; Middle Aged