AIM:To observe the efficacy of intravitreal conbercept injection for chronic central serous chorioretinopathy (CSC).METHODS: Nine eyes of 9 patients diagnosed as chronic CSC between October 2015 to May 2016 were treated with an intravitreal injection of conbercept (0.5mg/0.05mL) (six patients were given the same does of intravitreal injection again at 1mo after the first injection).Follow-up observation was at 1, 2, and 6mo after injection.Observed indicators included best-corrected visual acuity (BCVA), intraocular pressure, optical coherence tomography (OCT), fundus fluorescein angiography (FFA), choroidal indocyanine green angiography (ICGA), macular fovea thickness (CMT), subfoveal choroidal thickness (SFCT).RESULTS:Seven of the 9 patients responded significantly to the drug, while 2 patients had no response.The CMT was 373.12±72.43μm at baseline, which decreased significantly to 332.05±67.13μm, 282.24±62.30μm and 225.56±71.08μm at 1, 2 and 6mo after the intravitreal injection.The mean thickness of SFCT was 422.11±64.82μm before treatment.The choroidal thickness of non-responsive patients before treatment was below average, respectively 353μm and 365μm.The SFCT of 1, 2, and 6mo after treatment was 391.45±75.24μm, 365.53±63.07μm, 355.40±66.65μm.Before treatment and 1mo after, there was no significant difference (P=0.074), but there was statistically significant (P<0.01) between those of before and 2mo and 6mo after.The mean BCVA of the prior treatment was 0.53±0.32, the after treatment was 0.65±0.20, there was no different between the two(P>0.05).CONCLUSION: Intravitreal conbercept injection in chronic CSC may have some effect in accelerating subertinal fluid resolution and decreasing the CMT.The SFCT within 6mo after treatment was significantly lower than pretreatment.The SFCT may be an indicator of whether patients respond.

Objective To evaluate the functional changes of visual cortex (V1) in patients with primary open angle glaucoma (POAG) by fMRI retinotopic mapping technology. Methods Fifteen POAG patients and 15 healthy volunteers underwent stimulations with fMRI retinotopic mapping stimulus and contrast-reversing checkerboard patterns stimulus on a Siemens Trio 3.0 T MRI whole-body scanner for functional data collection. Comparisons of V1 fMRI responses between the glaucomatous eyes and the healthy eyes of the patients were carried out using paired samples t-test, while independent samples t-test was used to compare V1 fMRI responses and activations between the healthy eyes of patients and the age-, gender- and side- matched eyes of normal people. Differences of V1 cortical functions and visual functions were analyzed by linear correlation analysis when the glaucomatous and the healthy eyes were simulated individually. Results (1) V1 fMRI responses of the individually stimulated glaucomatous eyes[(1.24±0.72)%]were weaker than those of the healthy eyes[(2.18±0.93)%](t=4.757,P<0.01). Comparisons of V1 fMRI responses between the glaucomatous eyes and matched eyes of normal people, as well as between the healthy eyes of patients and the matched eyes of normal people, were performed respectively: the responses in the glaucomatous eyes[(1.24±0.72)%]were weaker than those in the matched eyes of normal people[(2.01±0.65)%](t=-3.011,P<0.01). There was no statistical difference of the responses between the healthy eyes from patients[(2.18±0.93)%]and the matched eyes of normal people[(1.95±0.75)%](t=0.742,P＞0.05). (2) Differences of V1 cortical functions were negatively correlated with those of visual functions in the individually stimulated glaucomatous and healthy eyes (r=-0.887, P<0.01). (3) The activated area indexes of V1 cortexes in the healthy eyes from patients (0.72±0.12) were lower than those in the matched eyes of normal people (0.85±0.09) (t=-3.801, P<0.01). Conclusion Cortical function impairment was in accordance with visual function impairment in glaucoma. Located and quantified measurement with fMRI retinotopic mapping was a useful method for clinical follow-up and evaluation of functional alteration of glaucomatous visual cortex, and a potentially useful means of studying trans-synaptic degeneration of visual pathways of in vivo glaucoma.

Background TLR-4 is a natural immunity receptors in immunity,and it plays an important role in the repair of central nervous system damage.But its effect in glaucoma optic nerve injury is unclear.Objective This study was to investigate the expression of TLR-4 in retina with high intraocular pressure(IOP)in genetic and Drotein level and therefore explore the mechanism of TLR-4 on retinal ganglion cells(RGCs)injury. Methods Chronic ocular hypertension models were established in the right eyes of 150 clean purebred Sprague-Dawley rats by cauterizing the 3 sallow sclera veins.IOP was measured before and after 2 h,1 day,3,7,14,28,56 days after operation by PEN Ⅱ TONO-type pen tonometer.The expression of TLR-4 protein in rat retina was detected by immunohistochemistry and Western blot,and expression of TLR-4 mRNA was assayed by real time-PCR.This experimental procedure foliowed the Statement of Association for Research in Vision and Ophthalmology. Results The IOP was elevated in various time points after operation in experimental group,showing significant differences in comparison with control group(P<0.01).The immunohistochemistry revealed that the expression of TLR-4 protein in rat retina with chronic hypertension in 2 h,1 day,3,7,14,28,56 days after operation with the high A298 values in comparison with control eyes(P<0.05-0.01).Increased levels of TLR-4 mRNA in rat retinas were detected by RTPCR in high IOP eyes compared with control eyes in all time points after operation,presenting statistically significant differences between two groups(P<0.05-0.01).Western blot detection displayed the high expression of TLR-4 in retina in high IOP eyes early after operation with statistically significant results between model group and control group (P<0.05-0. 01). Conclusion TLR-4 is up-regulated in rat retina with chronic high IOP,suggesting that TLR-4 plays an immunoregulatory effect in glaucomatous eye.

Objective To evaluate the effects of the chronic Keshan disease's self-management in Liangshan Prefecture of Sichuan Province. Methods According to "The Serf Management Program of Chronic Keshan Disease in Liangshen Prefecture of Sichuan Province", 56 chronic Keshan disease patients were selected in the personalized self-manngemant evaluation under the instructions by endemic disease specialists and the rural doctors. Evaluation was based on changes of indexes such as the clinical symptoms, general health conditions, the electrocardiogram, X-ray, the heart function, etc, before treatment and 3 and 6 months following the treatment. Results Clinical signs and symptoms of the patients were significantly improved 3 and 6 months after the treatment, and the improvement was more obvious 6 months than 3 months following the treatment(P<0.05 or< 0.01). After treatment for 3 months, the patients'electrocardingram and heart function did not show obvious change (X2=0.05,039, P0.05); hut obvious improvements Eexcept X ray results(X2=0.61 ,P0.05)] were seen 6 months after treatment (X2=4.36,16.84, P<0.05 or<0.01). Altogether, among the 56 patients evaluated after treatment for 6 months, none achieved the clinical cure standard, 26 cases(46.3%) showed significant improvement, 17 cases (30.4%) were stable, 5 cases (8.9%) were aggravated,one case (1.8%) lost contact, and 6 cases (10.6%) died. Conclusion The project of the chronic Keshan disease's self-management is suitable for the present situati,on of the endemic regions and can he introduced to many places in our country.

OBJECTIVETo study the effect of heme oxygenase-1(HO-1) on proteins related to apoptosis in INS-1 cells with exposure to intermittent high glucose.

METHODSINS-1 cells cultured in vitro were divided into control group, persistent high glucose group (PHG), intermittent high glucose group (IHG), CoPP + intermittent high glucose group (CoPP+IHG), and ZnPP+ intermittent high glucose group (ZnPP+IHG). After 72 h of treatment with the corresponding protocols, the cells were examined for expressions of HO-1 protein by Western blotting and for expressions of Bax and Bcl-2 by immunocytochemistry.

RESULTSIn comparison with the control group, the cells in both PHG group and IHG group showed significantly increased expressions of HO-1 (P<0.01) and decreased Bcl-2/Bax ratios (P<0.05). The cells in CoPP+ IHG group exhibited a greater HO-1 protein expression but a lower Bcl-2/Bax ratio than those in IHG group (P<0.05) The ZnPP+IHG group demonstrated opposite changes in terms of HO-1, Bax and Bcl-2 expressions compared with the CoPP+IHG group.

CONCLUSIONIntermittent high glucose can lower Bcl-2/Bax ratio in INS-1 cells, and HO-1 may protect INS-1 cells against apoptosis possibly by up-regulating the Bcl-2/Bax ratio.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line ; Glucose ; administration & dosage ; adverse effects ; Heme Oxygenase (Decyclizing) ; metabolism ; Islets of Langerhans ; cytology ; drug effects ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo study of the role of nuclear transcription factor-κB (NF-κB) in high glucose-induced apoptosis in INS-1 cells.

METHODSRat insulinoma (INS-1) cells cultured in RPMI 1640 medium were treated with 11.1 mmol/L glucose, 33.3 mmol/L glucose, or 33.3 mmol/L glucose plus 5 µmol/L NF-κB inhibitors for 48 h. The expression of NF-κB subunit P65 protein in the cell nuclei was detected by Western blotting, IKK belta mRNA level by quantitative RT-PCR, and cell apoptosis by Annexin V-PI double staining.

RESULTSCompared with the control levels, IKK belta mRNA levels of the cells significantly increased in response to 33.3 mmol/L glucose exposure (P<0.01), which also resulted in significantly increased P65 protein expression in the cell nuclei (P<0.01) and cell apoptosis rate (P<0.05). Compared with those in the high glucose group, the expression of IKK belta mRNA and P65 protein and cell apoptosis rate decreased significantly after treatment with 33.3 mmol/L glucose plus 5 µmol/L NF-κB inhibitors (P<0.05).

CONCLUSIONHigh glucose induces NF-κB activation in INS-1 cells, and inhibition of NF-κB activation may protect INS-1 cells from high glucose-induced cell apoptosis.

Animals ; Apoptosis ; drug effects ; Cell Line, Tumor ; Gene Expression Regulation ; Glucose ; adverse effects ; metabolism ; Insulinoma ; pathology ; Pancreatic Neoplasms ; pathology ; Rats ; Transcription Factor RelA ; metabolism

OBJECTIVETo summarize the workflow, strategy and experience of prenatal genetic test for deafness based on the 6-year clinical practice.

METHODSThere were 213 families who received prenatal test from 2005 to 2011. Among the 213 families, 205 families had had one deaf child, including 204 couples with normal hearing and one couple of the deaf husband and normal wife, 8 families including 6 couples with normal hearing and 2 deaf couples, had no child before test. Genomic and mitochondrial DNA of each subject was extracted from whole blood. The etiology and recurrent risks in 212 families were confirmed by means of the genetic test of GJB2, SLC26A4 and mtDNA 12sRNA, but one family carried POU3F4 c.647G > A heterozygous mutation causing X-linked hereditary hearing impairment confirmed by pedigree study. The prenatal test was carried out during the pregnancy of all mothers from 11 to 30 weeks, and the following genetic information and counseling were supplied based on the results.

RESULTSThe recurrent risk was 25% in 209 families, including 204 families with one deaf child and 5 families without child, among which all couples were GJB2 or SLC26A4 mutation carriers and deaf children were caused by homozygous or compound GJB2/SLC26A4 mutations; The recurrent risk was 50% in 3 families, the father and his child in one family had compound SLC26A4 mutations and the mother with heterozygous SLC26A4 mutation, the wife had POU3F4 c.647G > A heterozygous mutation in another one family, and the husband with compound SLC26A4 mutations and the wife with mtDNA A1555G mutation and heterozygous SLC26A4 mutation simultaneously happened in the rest one family; The recurrent risk was 100% in one family of the deaf couple who were both found to carry homozygous or compound GJB2 mutations, and the deaf wife got pregnant by artificial insemination with the sperm from the local Human Sperm Bank. 226 times of prenatal test were applied in all 213 families that 11 families of them received prenatal test twice, and one family received three times. 46 times of prenatal testing showed that the fetuses carried parental mutations simultaneously or the same mutations with probands; while 180 times of prenatal test showed that the fetuses carried only one parental mutation or did not carry any mutation from parents. The following visit showed that all of these 180 families had given birth to babies who were all revealed to have normal hearing by new born hearing screening test.

CONCLUSIONSPrenatal diagnosis for deafness assisted by genetic test can provide efficient information about offspring's hearing condition, and the normative workflow and precise strategy highly guarantee the safe and favorable implementation of prenatal diagnosis.

Connexins ; genetics ; DNA Mutational Analysis ; DNA, Mitochondrial ; Deafness ; diagnosis ; genetics ; prevention & control ; Female ; Genetic Testing ; Heterozygote ; Humans ; Infant ; Pedigree ; Pregnancy ; Prenatal Diagnosis

BACKGROUNDPreviously we had successfully tracked adult human neural stem cells (NSCs) labeled with superparamagnetic iron oxide particles (SPIOs) in host human brain after transplantation in vivo non-invasively by magnetic resonance imaging (MRI). However, the function of the transplanted NSCs could not be evaluated by the method. In the study, we applied manganese-enhanced MRI (ME-MRI) to detect NSCs function after implantation in brain of rats with traumatic brain injury (TBI) in vivo.

METHODSTotally 40 TBI rats were randomly divided into 4 groups with 10 rats in each group. In group 1, the TBI rats did not receive NSCs transplantation. MnCl2·4H2O was intravenously injected, hyperosmolar mannitol was delivered to disrupt rightside blood brain barrier, and its contralateral forepaw was electrically stimulated. In group 2, the TBI rats received NSCs (labeled with SPIO) transplantation, and the ME-MRI procedure was same to group 1. In group 3, the TBI rats received NSCs (labeled with SPIO) transplantation, and the ME-MRI procedure was same to group 1, but diltiazem was introduced during the electrical stimulation period. In group 4, the TBI rats received phosphate buffered saline (PBS) injection, and the ME-MRI procedure was same to group 1.

RESULTSHyperintense signals were detected by ME-MRI in the cortex areas associated with somatosensory in TBI rats of group 2. These signals, which could not be induced in TBI rats of groups 1 and 4, disappeared when diltiazem was introduced in TBI rats of group 3.

CONCLUSIONIn this initial study, we mapped implanted NSCs activity and its functional participation within local brain area in TBI rats by ME-MRI technique, paving the way for further pre-clinical research.

Animals ; Brain Injuries ; physiopathology ; surgery ; Cell Movement ; Image Enhancement ; Magnetic Resonance Imaging ; methods ; Manganese ; Neural Stem Cells ; physiology ; transplantation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the functional changes of dendritic cells (DCs) after infection by recombinant retrovirus carrying human telomerase reverse transcriptase (hTERT) gene fragment.

METHODSInterleukin-12 (IL-12) levels in DC culture supernatant was determined by enzyme-linked immunosorbent assay (ELISA). The abilities of DCs infected with recombinant retrovirus carrying hTERT gene (hTERT-DCs) and non-infected DCs (N-DCs) to stimulate allogeneic lymphocyte proliferation were evaluated with mixed leukocytes reaction (MLR), and the surface markers of DCs including CD80, CD83, CD86 and HLA-DR were detected by flow cytometry. Cytotoxic T lymphocyte (CTL) assay was performed with CytoTox 96 non-radioactive cytoxicity assay.

RESULTSCompared with N-DCs, hTERT-DCs showed no significant changes in IL-12 secretion and capacity to stimulate allogeneic lymphocytes reaction, but had significantly lower CD83 expression. Specific CTLs induced by hTERT-DCs resulted in higher cytotoxicity against telomerase-positive target cells than that against the negative target cells.

CONCLUSIONInfection with the recombinant retrovirus carrying hTERT fragment may jeopardize the maturation of DCs, which, however, still retain their capacity to activate and stimulate lymphocyte proliferation and to prime autologous T lymphocytes to generate specific CTL against hTERT.

Cells, Cultured ; Dendritic Cells ; cytology ; immunology ; virology ; Genetic Vectors ; Humans ; Interleukin-12 ; biosynthesis ; Recombination, Genetic ; Retroviridae ; genetics ; metabolism ; T-Lymphocytes, Cytotoxic ; immunology ; Telomerase ; biosynthesis ; genetics

OBJECTIVETo study the relationship between the expression of serum amyloid A (SAA) and insulin resistance in 3T3-L1 adipocytes.

METHODS3T3-L1 adipocytes were incubated with different concentrations of dexamethasone (10, 100, and 1000 nmol/L) for 48 h to establish cell models of insulin resistance at different resistant levels (models 1, 2, and 3, respectively). The degree of insulin resistance of 3T3-L1 adipocytes was assayed by 2-deoxy-[(3)H]-D-glucose uptake. Semi- quantitative RT-PCR was performed for quantification of SAA mRNA expression. SAA concentrations in the culture medium were determined by ELISA.

RESULTDexamethasone did not affect the basal glucose transport (P>0.05). Insulin-stimulated glucose uptake was significantly decreased by 15% (P<0.05), 40% (P<0.01), and 55% (P<0.01) in models 1, 2, and 3 in comparison with the untreated group, respectively; the expressions of SAA mRNA were upregulated by 2.5 (P<0.01), 3.33 (P<0.01), and 4.08 folds (P<0.01) and SAA concentrations increased by 2.05, 3.13, and 4.23 folds, respectively. The expressions of SAA mRNA were positively correlated to the degree of insulin resistance (r=0.773, P<0.01) and SAA concentration (r=0.832, P<0.01).

CONCLUSIONA cell model of insulin resistance has been established in 3T3-L1 adipocytes by dexamethasone exposure. SAA is closely associated with insulin resistance and may serve as a marker of insulin resistance.

3T3-L1 Cells ; Adipocytes ; metabolism ; Animals ; Deoxyglucose ; metabolism ; Dexamethasone ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Insulin Resistance ; Mice ; RNA, Messenger ; genetics ; metabolism ; Serum Amyloid A Protein ; genetics ; metabolism