AIM: To investigate the influence and mechanism of blue light on the proliferation of human retinal pigment epithelial cells.METHODS: Cells were divided into two groups,including blue light group and control group.The 35W white light lamp with blue filter was used to establish damaged RPE cell model in vitro.Blue ray wavelength ranged between 470nm and 520nm.And the light intensity was about 2000Lx.After exposure to blue light,we tested the proliferation of human retinal pigment epithelial cells by CCK-8 kit.And then expression of miR-103 was measured by the real-time PCR.RESULTS: Exposure to blue light inhibited the proliferation of human retinal pigment epithelial cells and increased the expression of miR-103.Moreover,up-regulation of miR-103 inhibited the proliferation of human retinal pigment epithelial cells,and down-regulates miR-103 promoted the proliferation of human retinal pigment epithelial cells.CONCLUSION: Blue light inhibits the proliferation of human retinal pigment epithelial cells by the up-regulation of miR-103.

The authors helpfully discuss the design idea,experimental module design,examination methods,and experiment textbook construction in experimental teaching of microbiology,and conduct further researches on the basic skill training,verifying experiment,integrative experiment,and investigative experiment in the course. This study aims to enhance effects of the experimental teaching,to cultivate high potential talents who can master essential knowledge and skills,and creatively carry out scientific research.

OBJECTIVETo explore the functional brain localization with magnetic resonance imaging (MRI) after acupuncturing the Yuan-Source and He-Sea acupoints of Stomach Meridian of Foot-Yangming (ST).

METHODSThe study was performed in 30 healthy volunteers who underwent acupuncture at Yuan-Source acupoint (Chongyang, ST42) and He-Sea acupoint (Zusanli, ST36) (ST group). Ten of these were also underwent acupuncture at the non-acupoints as the control group. Blood oxygenation level dependent functional MRI was performed.

RESULTSIn the ST group, signal increasing areas were demonstrated in bilateral superior temporal gyri (Broadmann 22), bilateral supramarginal gyri (Broadmann 40), bilateral cerebellar hemispheres, bilateral cingulate gyri and isthmus of cingulate gyri (Broadmann 32, 30), bilateral superior parietal lobules (Broadmann 7); signal decreasing areas were shown in bilateral orbital gyri (Broadmann 11), bilateral temporal pole (Broadmann 38), right inferior frontal gyrus (Broadmann 47) and right medial occipitotemporal gyrus (Broadmann 36). In the control group, signal increases areas were demonstrated in superior temporal gyri, precentral gyri, cingulate gyri, thalamus, insula and cerebellum. The size, signal intensity and number of increasing areas in control group are less than in ST group.

CONCLUSIONCombined acupuncture of Yuan-Source and He-Sea acupoints of ST can activate and decrease the multiple brain regions of "splanchnic brain" and thus reach a new functional balance to relieve pain.

Acupuncture Points ; Adult ; Brain ; physiology ; Electroacupuncture ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Meridians ; Young Adult

Objective · To investigate the expression levels of estrogen receptor GPR30, elastin, collagen Ⅰ (Col Ⅰ ), and Col Ⅲ in the uterus round ligament and the cardial ligament in patients with pelvic organ prolapse (POP) and main components in extracellular matrix (ECM). Methods · 22 patients with POP (the POP group) and 10 patients with cervical intraepithelial neoplasia (CIN) Ⅲ (the control group) were enrolled. Hysterectomy specimens were collected and expressions of GPR30, elastin, Col Ⅰ and Col Ⅲ in the uterine round ligament and the cardial ligament tissues were detected with Western blotting and immunohistochemistry. Results · Western blotting and immunohistochemistry results showed that the expressions of GPR30, elastin, Col Ⅰ and Col Ⅲ in the uterine round ligament and the cardial ligament tissues were significantly lower in the POP group than in the control group. The differences were statistically significant (P<0.05). Conclusion · The occurrence of POP may be associated with estrogen receptor deficiency and decreased contents of collagen and elastin in relevant tissues. Whether GPR30 is involved in the synthesis, secretion and degradation of ECM major components in pelvic support tissues needs further investigation.

Objective To mvesugate the Tate of YMDD mutation accompamed with pre-core(region and core promotor region mutation and the clinical significance.Methods YMDD mutation and pre-core(at 1896 nu- cleotide)region and core promotor region(at 1762.1764 nucleotide)mutation were detected from the 122 patients with chronic hepatitis B virus after receiving lamivudine treatment above 6 months.Results 40 cases were tested for YMDI)mutations in 122 HBV patients with lamivudine treatment,and the positive rate of YMDD mutation was 32.8 %.After YMDD mutation,ALT,AST and HBV DNA of the patients significantly increased(P0.05).Conclusion The patients with YMDD mutation had higher rate of pre-core region(at 1896 nucleotide)and basal core promotor region(at 1762, 1764 nucleotide)mutation than those without YMDD mutation,but there was no correlation between the mutation and the deterioration of disease condition and the bad prognosis.

OBJECTIVETissue factor (TF) is an important factor in extrinsic coagulation. Tissue factor pathway inhibitor (TFPI) is a negative regulator of coagulation mediated by TF. Studies on TF and TFPI focus mainly on adult objects, seldom have been done on newborns, especially on sick newborns. The aim of this study was to observe the changes of TF and TFPI in plasma of newborns with infection jaundice and to research the effect of jaundice and infection on the balance of TF and TFPI in newborns.

METHODSThe content of TF and TFPI in plasma of 21 jaundiced newborns with infection and 8 jaundiced newborns without infection as control was determined quantitatively with the enzyme-linked immunosorbent assay (ELISA).

RESULTSThe content of TFPI and TF in plasma of jaundiced newborn with infection was significantly higher than that of controls [TFPI (21.0 +/- 4.3) vs. (16.2 +/- 1.9) microg/L, P < 0.01; TF (177 +/- 79) vs. (51 +/- 24) ng/L, P < 0.01]. The ratio of TFPI/TF was significantly lower in newborn with infection jaundice than the controls (137 +/- 61 vs. 319 +/- 67, P < 0.01). The 21 jaundiced newborns with infection were divided into the severe hyperbilirubinemia group (serum bilirubin > or = 205.2 micromol/L, n = 10) and the mild hyperbilirubinemia group (serum bilirubin < 205.2 micromol/L, n = 11). There was no significant difference of TFPI level between the severe hyperbilirubinemia group and mild hyperbilirubinemia group (P > 0.05). The TF content in the severe hyperbilirubinemia group was higher than that in the mild hyperbilirubinemia group (216 +/- 79 vs.141 +/- 63, P < 0.01), while the ration of TFPI/TF was lower in the severe hyperbilirubinemia group than in the mild hyperbilirubinemia group (100 +/- 30 vs. 171 +/- 74, P < 0.01).

CONCLUSIONInfection might induce imbalance between the coagulation inhibition and activation in newborns. Hyperbilirubinemia can aggravate the imbalance induced by the infection through increasing plasma TF level.

Bacterial Infections ; blood ; complications ; Case-Control Studies ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Infant, Newborn ; Jaundice, Neonatal ; blood ; etiology ; Lipoproteins ; blood ; Male ; Thromboplastin ; analysis

Although ethylene glycol (EG) has been widely used for embryo cryopreservation in domestic animals, few attempts were made to use this molecule to freeze mouse and human embryos. In the few studies that used EG for slow-freezing of mouse and human embryos, complicated protocols for human embryos were used, and the protocols need to be simplified. Besides, freezing mouse morula with EG as a cryoprotectant has not been reported. In this paper, we studied the effects of embryo stages, EG concentration, duration and procedure of equilibration, sucrose supplementation and EG removal after thawing on the development of thawed mouse embryos, using the simple freezing and thawing procedures for bovine embryos. The blastulation and hatching rates (81.92% +/- 2.24% and 68.56% +/- 2.43%, respectively) of the thawed late compact morulae were significantly (P < 0.05) higher than those of embryos frozen-thawed at other stages. When mouse late compact morulae were frozen with different concentrations of EG, the highest rates of blastocyst formation and hatching were obtained with 1.8mol/L EG. The blastulation rate was significantly higher when late morulae were equilibrated in 1.8 mol/L EG for 10 min prior to freezing than when they were equilibrated for 30 min, and the hatching rate of embryos exposed to EG for 10 min was significantly higher than that of embryos exposed for 20 and 30 min. Both rates of blastocyst formation and hatching obtained with two-step equilibration were higher (P < 0.05) than with one-step equilibration in 1.8 mol/L EG. Addition of sucrose to the EG-based solution had no beneficial effects. On the contrary, an increased sucrose level (0.4 mol/L) in the solution impaired the development of the frozen-thawed embryos. In contrast, addition of 0.1 mol/L sucrose to the propylene glycol (PG)-based solution significantly improved the development of the frozen-thawed embryos. Elimination of the cryoprotectant after thawing did not improve the development of the thawed embryos. The cell numbers were less (P < 0.05) in blastocysts developed from the thawed morulae than in the in vivo derived ones. In summary, embryo stage, EG concentration, duration and procedure of equilibration and sucrose supplementation had marked effects on development of the thawed mouse embryos, and a protocol for cryopreservation of mouse embryos is recommended in which the late morulae are frozen in 1.8 mol/L EG using the simple freezing and thawing procedures of bovine embryos after a two-step equilibration and the embryos can be cultured or transferred without EG removal after thawing.
Animals ; Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Dose-Response Relationship, Drug ; Embryo, Mammalian ; drug effects ; physiology ; Embryonic Development ; physiology ; Ethylene Glycol ; pharmacology ; Female ; Mice ; Morula ; physiology ; Pregnancy ; Sucrose ; pharmacology

OBJECTIVETo screen the gene expression profiles of IFN-alpha antiviral proteins based on a low-density cDNA Macroarray, and to explore the relationship between the expression of antiviral protein and the HBV replication.

METHODSThe HepG2 and HepG2.2.15 cells were treated with various concentrations of IFN-alpha (0 IU/ml, 100 IU/ml, 1000 IU/ml) of IFN-alpha for 6 h, and then the low-density cDNA Macroarray was used for analysing the expression profiles of antiviral genes and screening differential expressions of antiviral proteins. Meanwhile, the HepG2 cells were transiently transfected with HBV core protein-expressed plasmid pHBc-EGFP, and the expressions of antiviral proteins were analysed by RT-PCR assay. Moreover, the HepG2.2.15 cells were also transfected with the antiviral protein-expressed plasmid pcDNA3.1-Flag-MxA. ELISA was used for analysing the secreted HBV antigens, while dot blot and Southern blot were applied for analysing the extracellular HBV DNA and intracellular replicative intermediate HBV DNA in HepG2.2.15 cells. All data were presented as mean+/-SD and analyzed using the t-test and one-way analysis of variance (ANOVA) in the experiments.

RESULTSThe Macroarray results suggested that the expression of IFN-alpha antiviral genes like 6-16, IFITM1, IFITM2, IFITM3 and RING4 in HepG2.2.15 cells were partially inhibited. More importantly, it was found, in this research, the expression of antiviral protein MxA in HepG2.2.15 cells was completely suppressed. RT-PCR analysis indicated that the expression of MxA was also significantly decreased in HepG2 cells transfected with pHBc-EGFP plasmid. Although HepG2.2.15 cells transfected with pcDNA3.1-Flag-MxA plasmid could not inhibit extracellular HBV DNA and intracellular replicative intermediate HBV DNA, the MxA exerted some antiviral activities as it effectively suppressed the secretion of HBsAg and HBeAg in HepG2.2.15 cells.

CONCLUSIONSHBV and its antigen components probably influence the expression of antiviral proteins. IFN- resistance may be related to the down-regulation of antiviral proteins expression.

Antiviral Agents ; pharmacology ; Gene Expression Profiling ; Gene Expression Regulation, Viral ; Hep G2 Cells ; Hepatitis B virus ; drug effects ; physiology ; Humans ; Interferon-alpha ; pharmacology ; Plasmids

OBJECTIVETo investigate the effects of preweaning enrichment on the expression of activity-regulated cytoskeletal protein (Arc), an immediate early gene, and on the long-term memory in rats.

METHODSForty neonatal Sprague-Dawley rats were randomly assigned to control group (standard environment, n=20) and experimental group (enriched environment, n=20). The experimental group received enriched environment exposure from postnatal day 10 until weaning (2 weeks, 20 minutes per day). The open field and novel object recognition tests were performed at postnatal day 28. Arc expression was detected by Western blotting and immunohistochemistry.

RESULTSThere was no significant difference in the open field test between the two groups. However, in the novel object recognition test, the experimental group rats performed significantly better than the control rats after 1 and 24-hr retention. The preference index in the experimental group after 1-hr (59.61%+/-9.61% vs 50.46%+/-9.34%; P<0.05) and 24-hr retention (62.72%+/-14.12% vs 52.39%+/-9.16%; P<0.05 ) was significantly higher than that in the control group. Arc expression in both areas CA1 and DG of hippocampus in the experimental group increased significantly compared with that in the control group (P<0.01).

CONCLUSIONSPreweaning enrichment can up-regulate the expression of immediate early gene, Arc, in the hippocampus of the rats, and promote their long-term memory.

Animals ; Cytoskeletal Proteins ; analysis ; Female ; Hippocampus ; chemistry ; physiology ; Immunohistochemistry ; Male ; Memory ; Nerve Tissue Proteins ; analysis ; Rats ; Rats, Sprague-Dawley

BACKGROUNDNon-adhesive liquid embolic agents are increasingly gaining importance in the embolization of cerebral arteriovenous malformations (AVMs). We investigated the use of poly (N-isopropylacrylamide) (PNIPAM) as a non-adhesive embolic agent in swine rete mirabile.

METHODSThe PNIPAM hydrogel was mixed with iohexol and embolization was performed in swine rete mirabile in 30 animals. The microcatheter was examined after embolization. Follow-up angiography was performed for embolic efficacy after embolization. Embolized retia were examined histopathologically, and the alterations of inside rete and surrounding tissue were observed.

RESULTSThe copolymer hydrogel was used for rete embolization in 30 swine, 28 swine survived the procedure, 2 swine died, 1 swine died of cerebrum infarction and the other died of embolic agent reflux into the occipital artery. The inside wall of the microcatheter was smooth, without copolymer adhering to it. Follow-up angiography was performed in 22 swine, there was no rete recanalization in 20 swine and partial rete recanalization in 2 swine because of the trunk embolization of ascending pharyngeal arteries. Histopathologically, the copolymer was found diffused into vessels of 100 - 150 microm in diameter. In acute group, neutrophils scattered surrounding the copolymer and endothelial integrity was observed, without endothelial denuding and necrosis. In subacute and chronic groups, the copolymer was found inside retia, a few mononuclear cells and eosinocytes scattered inside and surrounding it. The muscular layer was loosened with most muscular nuclei degraded.

CONCLUSIONExperimental rete embolization with PNIPAM, made radiopaque with iohexol, is technically feasible in swine. Because of its properties, PNIPAM has great potential as a therapeutic non-adhesive embolic agent.

Acrylic Resins ; Animals ; Embolization, Therapeutic ; methods ; Intracranial Arteriovenous Malformations ; therapy ; Swine