Objective To study the regulation of microRNA 199a (miR-199a) on adhesion,migration and invasion ability of human eutopic endometrial stromal cells (ESC) from patients with endometriosis.Methods ESC were transfected with miR-199a mimics or negative control (NC) RNA by lipofectamine 2000.The adhesion,migration and invasion ability of ESC were detected by cell adhesion assay,scratch assay,cell migration assay and matrigel invasion assay,respectively.Luciferase reporter assay was used to evaluate whether IKKβ was the target gene of miR-199a.The expression of ikappa B kinase beta (IKKβ),inhibitory kappa B alpha (IκB-α),phospho-IκB-α (p-IκB-α) and nuclear factor-kappa B (NF κB) protein were measured by western blot.Results ( 1 ) Adhesion potential:the adhesion inhibitory rates were ( 14 ± 4 )％ in miR-199a group and 0 in control group,which showed significant difference (P＜0.01 ).(2) Migration and invasion:in the scratch assay,ESC transfected with miR-199a exhibited a lower scratch closure rate than that of controls.In migration and invasion assays,the migration and invasion ability of miR-199a group were significantly decreased compared with those of NC group [ 130 ± 31 vs.247±36 (P＜0.01); 63 ± 15 vs.133 ± 17 (P＜0.01),respectively].(3) The luciferase activity of miR-199a group was significantly lowered than that of control group [ 0.160 ± 0.006 vs.0.383 ± 0.083 ( P ＜0.01 ) ].The protein levels of IKKβ,p-IκB-α,IκB-α and NF-κB of 0.350 ±0.195,0.443 ±0.076,1.970 ±0.486 and 0.454 ± 0.147 in miR-199a group were significantly different compared with the NC group in which the protein levels were set at 1.000 ( P ＜ 0.01 ).Conclusions miR-199a can inhibit the adhesion,migration and invasion of the ESC.IKKβ is the target gene of miR-199a in ESC.One of the mechanisms of the inhibition effect is probably that miR-199a inhibits the activation of NF-κB signaling pathway by targeting IKKβ gene.

Literatures on knee osteoarthritis treated by acupuncture both in China and abroad published in the mainstay periodicals in recent 10 years were selected, and analyses were done in the following aspects: (1) Randomization, (2) Control group, (3) Sample size, (4) Intervention measurements, (5) Intervention periods, (6) Evaluation on therapeutic effects, (7) Follow-up assessment, (8) Adverse effects, (9) Ratio of the lost case. The result indicates that differences can still be found on the trial designation in China and abroad. The domestic research design should be more comprehensively and strictly.
Acupuncture Therapy ; adverse effects ; China ; Clinical Trials as Topic ; Humans ; Osteoarthritis, Knee ; therapy

OBJECTIVETo explore the synergic effect of heparan sulfate (HS) and cytokines on the growth of human umbilical cord blood (UCB) hematopoietic cells.

METHODSHematopoietic cells from human UCB were cultured with (1) cytokines (rhIL-3, rhIL-1beta, rhIL-6 and SCF) or (2) SN (supernatant from cultured human marrow stromal cells) and cytokines, or (3) SN, or (4) HS and cytokines. Cellular proliferation, CFU-GM yields and changes of cell immunophenotype were observed.

RESULTSHematopoietic cell proliferation reached peak at the 14th day. The number of total nucleated cells increased 134.5-, 171.3-, 81.5- and 167.2-fold in (1), (2), (3) and (4) groups, respectively, at the 21th day, the (3) group significantly decreased. CD(34)(+) cells increased at the 7th day and reached peak, at the 14th day with a percentage of 68.4%, 82.5%, 69.8% and 79.3%, and at the 21th day 56.2%, 71.7.%, 12.3% and 73.3%, respectively. CD(33)(+) cells reached peak at the 14th day and increased by 80.2%, 68.6%, 81.6% and 70.3%, respectively, and remained these levels at the 21th day. CD(38)(+) cells increased by 66.6%, 73.8%, 70.4% and 71.9% at the 7th day and remained this level at the 14th and the 21th day. From the first week of culture, the percentage of CFU-GM increased in all of the four groups, at the second week of culture, it increased by 250%, 279%, 217% and 273%, and at the third week still increased by 151%, 240%, 145% and 231%, respectively.

CONCLUSIONThe combination of SN and cytokines have a synergic effect in promoting proliferation of hematopoietic cell from human UCB. The synergic effect remained the same when SN was replaced by heparan sulfate.

ADP-ribosyl Cyclase ; analysis ; ADP-ribosyl Cyclase 1 ; Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Cell Division ; drug effects ; Cells, Cultured ; Cytokines ; pharmacology ; Drug Synergism ; Fetal Blood ; cytology ; drug effects ; immunology ; Heparitin Sulfate ; pharmacology ; Humans ; Infant, Newborn ; Interleukin-1 ; pharmacology ; Interleukin-3 ; pharmacology ; Leukocytes, Mononuclear ; cytology ; drug effects ; immunology ; Membrane Glycoproteins ; Recombinant Proteins ; pharmacology ; Sialic Acid Binding Ig-like Lectin 3 ; Time Factors

Abstract?AIM: To evaluate the visual acuity, visual function, q uality of lfi e, and cost/effectiveness of cataract surgery in patients with low soc ioeconomics tatus ( LSES ) who received government subsidies for free cataract surgery.?METHODS: Retrospectively, 334 eyes of 334 patient s were reviewed. Visual function and quality of l ife questionnaires we re available in 83 patie nts. The cost/effectiveness of cataract surgery was analyzed in 225 patients with LSES and 12 0 patients who rec eived standard cataract surgery. Mutl iple linear regression anal ysis and logistic regression analysis were used to assess the factors affecting the postoperative visual acuity.?RESULTS: At 1wk postoperatively, uncorrected visual acuity ( UCVA ) was improved significantly compared to these preoperait vely (P<0.50 ), 235 eyes (70.4%) had UCVA of 20/40 or better, and 16 eyes (4.8%) had UC VA wor se than 20/400. Multiple linear regresis on ana lysis revealed that other ocular diseases and cataract classification wer e factors contributing to po stop erative UC VA. Logistic regression analysis showed that low preoperative visual ac uti y and other ocular diseases were risk factors of postopera tive low vision, and the odds ratio (95%confidence int erval) values were 6.5 (2.8-14.8) and 1.3 (1.0-1.6), respe ctively.Visual fun ction and quality of life were significantly improved after the surgery. Th e cost/effetc iveness of c ataract su rgery in patients with LSES was lower than that in standard cataract patients.?CONCLUSION: Visual acuity and visual function were significantly better after the surgery in catara ct patients wti h LSES, and the operation cost was low.Quality of life in these patients was significantly improved.

OBJECTIVETo evaluate the clinical efficacy and efficacy sustainable time of acupuncture in knee osteoarthritis (KOA).

METHODSThe non-randomized concurrent control trial was adopted. One hundred and ninety-three cases of KOA were divided into an immediate acupuncture group (group A, 97 cases) and a delayed acupunc-weeks at the end of treatment. In group B, the same acupuncture therapy was applied after waiting 4 weeks. The acupoints in the two groups were Liangqiu (ST 34), Dubi (ST 35), Zusanli (ST 36), Yanglingquan (GB 34), Yinlingquan (SP 9), Xuehai (SP 10), Xiyan (EX-LE 4), Xiyangguan (GB 33). WOMAC (Western Ontario and McMasters Universities Osteoarthritis) was used for the assessment of the primary index and VAS (visual analogue scale) was for the secondary index. The evaluation was accomplished by the patients at the beginning of trial, on the 4th and 8th weeks. In each group, 72 patients finished the trial and the data of the lost cases were included in the final data analysis.

RESULTSIn the 4th week of trial, WOMAC score was (25. 8+/-22.0) in group A difference (P<0. 001). VAS scorewas (31. 8+/-24. and was (43.8+/-22.2) in group B, indicating the significant 6) in group A and was (56. 6 +/-25. 8) in group B, indicating very significant difference (P<0. 001). In the 8th week, the efficacy was reduced slightly in the follow-up of group A, but it was improved apparently as compared Acupuncture relieves joint pain and improves joint function obviously.by th patiĩeffr,a Mtaetfti-?an tf ri-with that before treatment.

CONCLUSIONAcupuncture relieves joint pain and improves joint function obviously.The effect of acupuncture is still sustainable in 4 weeks after terminating the treatment.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Arthralgia ; therapy ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; therapy ; Treatment Outcome

Rett syndrome (RTT) is a disorder characterized by regression of spoken language and hand use,distinctive hand stereotypies,accompanying with severe psychomotor developmental retardation and retrogression.RTT becomes recognizable at 6-18 months and female are absolutely susceptive.MECP2 mutations are closely related to the development of RTT.Revised diagnostic criteria for RTT (2010) ensure a high degree of homogeneity in cases enrolled in treatment and clinical studies throughout the world.As for the treatment,no crucial advancement has been clinically applied recently,but some valuable basic research is in progress.This paper reviews the genetic research,clinical diagnosis and treatment of RTT,and promotes understanding of the new diagnostic criteria and basic research.

OBJECTIVETo investigate the effect of p65 gene inhibited by siRNA on neuronic differentiation in the marrow mesenchymal stem cells (MSCs).

METHODSThe MSCs were transfected with Rn-p65-siRNA. Fasudil hydrochloride induced MSCs differentiating into neurons. The non-transfected group and negative control group (transfected with negative control siRNA marked by Cy3) were used as controls. The fluorescence expressed by transfected MSCs were observed under inverted fluorescence microscope at 24 h,48 h and 72 h after transfected with negative control siRNA. The viability of MSCs was detected by MTT at 24 h, 48 h and 72 h after transfected with Rn-p65-siRNA. The expressions of p65 mRNA and protein in MSCs were detected by RT-PCR and Western blot respectively. The expressions of p65 protein, NSE, MAP-2 and glial fibrillary acidic protein (GFAP) were detected by immunocytochemical method after transfection for 6 h.

RESULTSThe fluorescence of MSCs was mostly displayed after transfection of 72 hours and the efficiency of transfection was up to 83.3% ± 3.8%. Meanwhile, the p65 mRNA and p65 protein expressed by MSCs of transfected group were significantly decreased (P < 0.05); MTT displayed that the viability of MSCs was also significantly reduced (P < 0.05). The best efficiency of induction was observed in the transfected group. There were higher expressions of NSE and MAP-2 than the other group (P < 0.05).

CONCLUSIONThe p65 gene inhibited by siRNA can promote the marrow mesenchymal stem cells to differentiate into neurons.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine ; analogs & derivatives ; Animals ; Cell Differentiation ; Glial Fibrillary Acidic Protein ; metabolism ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; RNA, Messenger ; RNA, Small Interfering ; Rats ; Transcription Factor RelA ; antagonists & inhibitors ; metabolism ; Transfection

OBJECTIVETo investigate the effects of dexamethasone on expression of matrix metalloproteinase-9 (MMP-9) in rats with acute lung injury induced by phosgene.

METHODSThe rats were randomly divided into 3 groups: normal control group that consists of the rats with air exposure, phosgene group that consists of the rats with phosgene exposure and dexamethasone group that consists of the rats with phosgene exposure after 2.5 mg/kg dexamethasone being injected. Wet and dry ratio of the lung (W/D) was calculated, and leukocyte count and total protein content of bronchoalveolar lavage fluid (BALF) were recorded at 2 h after exposure. The concentrations of MMP-9 in the serum and BALF were measured by enzyme-linked immunosorbent assay. The pathologic changes of lung tissues were observed under light microscopy. The immunohistochemistry and the RT-PCR were used to detect the contents of MMP-9 in the lung tissue.

RESULTSCompared with phosgene group, the lung W/D, protein content and WBC count in of BALF dexamethasone group was significantly decreased (P < 0.01). MMP-9 levels of the serum and BALF in dexamethasone group were (4.799 +/- 0.043) microg/L and (15.052 +/- 0.029) microg/L, respectively, which were significantly lower than those [(9.439 +/- 0.100) and (20.640 +/- 0.446) microg/L] in phosgene group (P < 0.01). Compared with phosgene group (2.789 +/- 0.282),the expression level (1.183 +/- 0.260) of lung MMP-9 mRNA in dexamethasone group was significantly lower than that in phosgene group (P < 0.01). Histological experimental results showed the marked hyperemia and thickening of alveolar walls and stroma cells infiltrating and more visible alveolar structure damage of alveolar walls in phosgene group while the alveolar structure and the alveolar walls were clear and slightly thickened with inflammatory cells in dexamethasone group. Immunohistochemical results showed that MMP-9 protein expression levels of lung and bronchus tissues in normal control group and dexamethasone group were weakly positive, which in phosgene group were strongly positive.

CONCLUSIONDexamethasone has a beneficial effects on acute lung injury induced by phosgene in rats due to the inhibiting MMP-9.

Acute Lung Injury ; chemically induced ; drug therapy ; metabolism ; Animals ; Bronchoalveolar Lavage Fluid ; Dexamethasone ; therapeutic use ; Disease Models, Animal ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Phosgene ; toxicity ; Rats ; Rats, Sprague-Dawley

A new sisomicin resistance gene sisR was cloned from sisomicin-producing Micromonospora inyoensis. The sisR fragment was obtained by PCR amplification. The primer pairs were designed based on grm gene sequence from gentamicin-producing Micromonospora purpurea. The template DNA was isolated from Micromonospora inyoensis. A series of different DNA fragments were amplified by PCR, which were sub-cloned to vector pUC19 for further identification. It was found that five specific transformants containing target DNA fragments could resist high concentrations of sisomicin (over 1000 microg/mL sisomicin). One of them designated as sisR, was then sequenced and the alignment among sisR and other related genes showed that sisR gene differs from any known genes. It was concluded that sisR gene is a sequence that has not been reported so far.
Anti-Bacterial Agents ; pharmacology ; Bacterial Proteins ; genetics ; Base Sequence ; Cloning, Molecular ; Drug Resistance, Microbial ; genetics ; Genes, Bacterial ; Micromonospora ; genetics ; Molecular Sequence Data ; Sisomicin ; pharmacology

OBJECTIVETo investigate the clinical features of Epstein-Barr virus-related hemophagocytic lymphohistiocytosis (EBV-HLH), to analysis the outcome of HLH-2004 protocol, and to explore the prognostic factors in EBV-HLH patients.

METHODSThe clinical features at onset and outcome of HLH-2004 protocol from 83 pediatric patients with EBV-HLH enrolled from January 2006 to December 2009 in our hospital were analyzed retrospectively. Univariate and multivariate COX regression analysis were used to identify statistically significant prognostic factors.

RESULTS(1) Among the 83 patients, 45 were males and 38 were females. The age of onset ranged from 6 months to 14 years 4 months. 44 patients were treated with HLH-2004, and 3-year overall survival (OS) was (55.8 ± 7.9)%. (2) The most common clinical features of EBV-HLH included high fever, cytopenia, hepatosplenomegaly, and coagulopathy; The respiratory symptoms, angina phlogistic, skin rashes, neurologic abnormality were rare. 97.3% of patients showed an elevation of serum ferritin, liver dysfunction and lipid metabolism disorders was found in most of EBV-HLH patients. 89.0% of patient had hemophagocytosis in bone marrow at diagnosis of EBV-HLH. (3) COX regression analysis revealed that anemia degree, serum albumin < 30 g/L, CD4:CD8 abnormity, NK cell < 3%, treatment protocol were related with the prognosis significantly (P < 0.05).

CONCLUSIONEBV-HLH in pediatric patients has severe clinical feature and poor prognosis. HLH-2004 protocol is an effective treatment for patients with EBV-HLH. Symptomatic treatment can't rescue the patients of EBV-HLH.

Adolescent ; Child ; Child, Preschool ; Epstein-Barr Virus Infections ; drug therapy ; Female ; Herpesvirus 4, Human ; Humans ; Infant ; Killer Cells, Natural ; Lymphohistiocytosis, Hemophagocytic ; drug therapy ; virology ; Male ; Prognosis ; Retrospective Studies ; Treatment Outcome