Plasma metabolomic profiling of proliferative diabetic retinopathy.

BACKGROUND: Proliferative diabetic retinopathy (PDR), a sight-threatening retinopathy, is the leading cause of irreversible blindness in adults. Despite strict control of systemic risk factors, a fraction of patients with diabetes develop PDR, suggesting the existence of other potential pathogenic factors underlying PDR. This study aimed to investigate the plasma metabotype of patients with PDR and to identify novel metabolite markers for PDR. Biomarkers identified from this study will provide scientific insight and new strategies for the early diagnosis and intervention of diabetic retinopathy. METHODS: A total of 1024 patients with type 2 diabetes were screened. To match clinical parameters between case and control subjects, patients with PDR (PDR, n = 21) or those with a duration of diabetes of ≥10 years but without diabetic retinopathy (NDR, n = 21) were assigned to the present case-control study. Distinct metabolite profiles of serum were examined using liquid chromatography-mass spectrometry (LC-MS). RESULTS: The distinct metabolites between PDR and NDR groups were significantly enriched in 9 KEGG pathways (P < 0.05, impact > 0.1), namely, alanine, aspartate and glutamate metabolism, caffeine metabolism, beta-alanine metabolism, purine metabolism, cysteine and methionine metabolism, sulfur metabolism, sphingosine metabolism, and arginine and proline metabolism. A total of 63 altered metabolites played important roles in these pathways. Finally, 4 metabolites were selected as candidate biomarkers for PDR, namely, fumaric acid, uridine, acetic acid, and cytidine. The area under the curve for these biomarkers were 0.96, 0.95, 1.0, and 0.95, respectively. CONCLUSIONS: This study suggested that impairment in the metabolism of pyrimidines, arginine and proline were identified as metabolic dysregulation associated with PDR. And fumaric acid, uridine, acetic acid, and cytidine might be potential biomarkers for PDR. Fumaric acid was firstly reported as a novel metabolite marker with no prior reports of association with diabetes or diabetic retinopathy, which might provide insights into potential new pathogenic pathways for diabetic retinopathy.

[Expression and significance of Shh and Wnt5a genes in Cornelia de Lange syndrome].

OBJECTIVE: To study the expression of Shh and Wnt5a genes in the limb buds of NIPBL+/- fetal rats and the association of these two genes with Cornelia de Lange syndrome (CdLS). METHODS: A total of 72 NIPBL+/- fetal rats were divided into an experimental group and a control group, with 36 rats in each group. The limb buds were collected from 12 fetal rats each on embryonic days 10, 11 and 12 (E10, E11 and E12) respectively. Real-time PCR and Western blot were used to measure the mRNA and protein expression of Shh and Wnt5a. RESULTS: The mRNA and protein expression of Shh and Wnt5a was detected in the limb buds on E10, E11 and E12, and the experimental group had significantly lower expression than the control group (P<0.01). The mRNA and protein expression of Shh and Wnt5a in limb buds was at a low level on E10, followed by an increase on E11 and a reduction on E12, and the expression on E12 was still lower than that on E10 (P<0.01). CONCLUSIONS: The mRNA and protein expression of Shh and Wnt5a are consistent. The pathogenesis of CdLS may be associated with the low mRNA and protein expression of Shh and Wnt5a inhibited by the low expression of NIPBL gene.

[Relationship between soil bacterial community and environmental factors in the degraded alpine grassland of eastern Qilian Mountains, China]. / ä¸œç¥è¿žå±±é€€åŒ–é«˜å¯’è‰åœ°åœŸå£¤ç»†èŒç¾¤è½ä¸ŽåœŸå£¤çŽ¯å¢ƒå› å­é—´çš„ç›¸äº’å ³ç³».

To explore the distribution of bacterial community and its relationship with soil environmental factors in degraded alpine grasslands in the eastern Qilian Mountains, we analyzed the changes of bacterial community structure and diversity across lightly, moderately and severely degraded grasslands by using high-throughput sequencing technology. Redundancy analysis (RDA) was performed to analyze the relationship between soil bacterial communities and soil environmental factors by using CANOCO 4.5 software. The results showed that there were significant differences in soil physicochemical properties among different degraded alpine grasslands. There were 257125 effective sequences, 180826 high-quality sequences and 4790 OTUs. The Chao1 index was lightly degraded grassland > moderately degraded grassland > severely degraded grassland; Shannon index was lightly degraded grassland > severely degraded grassland > moderately degraded grassland. Phylogenetic analysis indicated that the soil bacterial groups of each plot belonged to 33 phyla, with Actinomycetes, Proteobacteria and Firmicutes being the dominant groups in the three grasslands. From analysis of the proportion of soil bacteria in different degraded grassland, we found that the Actinomycetes, Acidobacteria and Proteobacteria increased first and then decreased with the degree of degradation, and the Firmicutes showed an opposite trend. The results of RDA analysis showed that the dominant groups of bacteria were significantly correlated with invertase, cellulase, phosphatas, pH, electronic conductivity, available nitrogen and available potassium. It was concluded that there were significant differences in soil bacterial communities among different degraded alpine grasslands in the eastern Qilian Mountains, and the soil environmental factors were the important factors driving the distribution of soil bacterial communities.

[Fabrication and accuracy research on 3D printing dental model based on cone beam computed tomography digital modeling].

OBJECTIVE: The aim of this study is to build a digital dental model with cone beam computed tomography (CBCT), to fabricate a virtual model via 3D printing, and to determine the accuracy of 3D printing dental model by comparing the result with a traditional dental cast. METHODS: CBCT of orthodontic patients was obtained to build a digital dental model by using Mimics 10.01 and Geomagic studio software. The 3D virtual models were fabricated via fused deposition modeling technique (FDM). The 3D virtual models were compared with the traditional cast models by using a Vernier caliper. The measurements used for comparison included the width of each tooth, the length and width of the maxillary and mandibular arches, and the length of the posterior dental crest. RESULTS: 3D printing models had higher accuracy compared with the traditional cast models. The results of the paired t-test of all data showed that no statistically significant difference was observed between the two groups (P>0.05). CONCLUSIONS: Dental digital models built with CBCT realize the digital storage of patients' dental condition. The virtual dental model fabricated via 3D printing avoids traditional impression and simplifies the clinical examination process. The 3D printing dental models produced via FDM show a high degree of accuracy. Thus, these models are appropriate for clinical practice.

[The receptor-dependent and non-receptor-dependent mechanism of melatonin activated BKCa channels in middle cerebral artery myocyte].

OBJECTIVE: To investigate the mechanisms through which myocyte large-conductance Ca2+-activated K+ (BKCa) channels mediate the vasodilation effects of melatonin on cerebral arteries (CAs). METHODS: Middle cerebral arteries (MCA) were obtained from 8-week-old male Wistar rats after anaesthetized. Middle cerebral arterial smooth muscle cells were enzymatically isolated. Whole cell recording mode of patch clamp technique was used to measure the current density of BKCa channel and voltage-gated potassium (KV) channel before and after adding melatonin. Currents density of melatonin on BKCa channels with melatonin receptor inhibitor 2-phenyl-N-acetyl (luzindole) was recorded using whole cell recording mode and open probability (Po) was recorded using single-channel attached recording mode. The conductance (G) and average open time (To) and off time (Tc) of the BKCa channel were detected before and after the addition of melatonin in the internal-outward mode. RESULTS: ① Melatonin markedly increased the whole-cell BKCa channel current density but not the voltage-gated potassium (KV) channel current density. ② Luzindole (1 µmol/L) greatly suppressed melatonin-induced increase of BKCa channel current density. ③ The Po of BKCa channel was significantly increased by melatonin (100 µmol/L) under cell attached recording mode, which was markedly inhibited by luzindole (1 µmol/L). ④ In inside-outside recording mode, melatonin (1 µmol/L, 100 µmol/L) reduced both To and Tc of BKCa channel, and Tc was reduced much more than To. CONCLUSIONS: Melatonin mediates vasodilation of MCA through the activation of BKCa channels via both melatonin receptor dependent and independent mode.

[Association between high-mobility group box 1 and neonatal respiratory distress syndrome].

OBJECTIVE: To study the association between serum level of high-mobility group box 1(HMGB1) and neonatal respiratory distress syndrome (NRDS). METHODS: A total of 35 infants with NRDS and 35 normal neonates (control group) were enrolled. Peripheral venous blood samples were collected with 12-24 hours after birth. ELISA was used to measure the serum level of HMGB1. RESULTS: The infants with mild and severe NRDS had a significantly higher serum level of HMGB1 than the control group (P<0.05). The infants with severe NRDS had a significantly higher serum level of HMGB1 than those with mild NRDS (P<0.05). The infants with NRDS who died had a significantly higher serum level of HMGB1 than those who survived (P<0.05). The receiver operating characteristic (ROC) curve showed that the optimal cut-off value for serum level of HMGB1 to predict NRDS was 625.3 pg/mL with an area under the ROC curve (AUC) of 0.846 (95%CI: 0.755-0.936), and the optimal cut-off value for serum level of HMGB1 to predict the death of infants with NRDS was 772.2 pg/mL with an AUC of 0.916 (95%CI: 0.813-1.000). CONCLUSIONS: Infants with NRDS have a significant increase in the serum level of HMGB1, and the serum level of HMGB1 can well predict the development and prognosis of NRDS.

[Diagnostic values of urinary netrin-1 and kidney injury molecule-1 for acute kidney injury induced by neonatal asphyxia].

OBJECTIVE: To investigate the values of urinary netrin-1 and kidney injury molecule-1 (KIM-1) in the early diagnosis of acute kidney injury (AKI) induced by neonatal asphyxia. METHODS: A total of 80 full-term neonates with asphyxia were enrolled (mild asphyxia: 34 neonates; severe asphyxia: 46 neonates). Forty normal full-term neonates were selected as the control group. Urinary samples were collected from the neonates in the three groups within 12 hours and 13-48 hours after birth. ELISA was applied to measure urinary levels of netrin-1 and KIM-1. Peripheral venous blood samples were also collected to measure serum creatinine (Scr) level. RESULTS: Compared with the control group, the asphyxia group had significantly higher urinary levels of netrin-1 and KIM-1 within 48 hours after birth and a significantly higher Scr level within 13-48 hours after birth (P<0.05). The neonates in the AKI group had significantly higher urinary levels of netrin-1 and KIM-1 and Scr level within 48 hours after birth than those in the non-AKI group (P<0.05). The areas under the receiver operating characteristic curve for urinary netrin-1 and KIM-1 levels within 12 hours after birth to predict AKI after asphyxia were 0.878 (95% CI: 0.775-0.981; P<0.01) and 0.899 (95% CI: 0.829-0.969; P<0.01), respectively. Any two indicators of urinary netrin-1 level, urinary KIM-1 level, and Scr level within 12 hours after neonatal asphyxia had a positive correlation (P<0.05). CONCLUSIONS: Urinary netrin-1 and KIM-1 levels increase significantly when neonates with asphyxia develop AKI. Urinary netrin-1 and KIM-1 can be used as indicators for the early diagnosis of AKI after asphyxia.

Identification of Salt-Stress-Induced Genes from the RNA-Seq Data of Reaumuria trigyna Using Differential-Display Reverse Transcription PCR.

Next generation sequencing (NGS) technologies have been used to generate huge amounts of sequencing data from many organisms. However, the correct choice of candidate genes and prevention of false-positive results computed from digital gene expression (DGE) of RNA-seq data are vital when using these genetic resources. We indirectly identified 18 salt-stress-induced Reaumuria trigyna transcripts from the transcriptome sequencing data using differential-display reverse transcription PCR (DDRT-PCR) combined with local BLAST searches. Highly consistent with the DGE results, the quantitative real-time PCR expression patterns of these transcripts showed strong upregulation by salt stress, suggesting that these genes may play important roles in R. trigyna's survival under high-salt environments. The method presented here successfully identified responsive genes from the massive amount of RNA-seq data. Thus, we suggest that DDRT-PCR could be employed to mine NGS data in a wide range of applications in transcriptomic studies. In addition, the genes identified in the present study are promising candidates for further elucidation of the salt tolerance mechanisms in R. trigyna.

Cloning and expression of a novel antifreeze protein AFP72 from the beetle Tenebrio molitor.

A novel antifreeze protein AFP72 cDNA (GenBbank accession No. AY929389) was obtained by RT-PCR from Tenebrio molitor. The 216 bp fragment encodes a protein of 72 amino acid residues. Sequence analysis revealed that the cDNA displays a high degree of homology with T. molitor antifreeze proteins, ranging up to 90.78%. Recombinant plasmids pMAL-p2X-afp72 and pMAL-c2X-afp72 were transferred into E. coil TBI to induce a MBP fusion protein by IPTG. The target fusion protein was released from the periplasm and cytoplasm by the cold osmotic shock procedure and sonication respectively. The content of the fusion protein came up to 38.9 and 41.5% of the total dissolved protein, respectively. The fusion protein was purified through an amylose affinity column, and incised by factor Xa. Molecular sieve chromatography was used to achieve a high state of purity of the target protein. The purified target protein displayed a single band in SDS-PAGE. The fusion protein was shown to increase resistance to low temperatures in bacteria. This finding could help in further investigations of the properties and function of antifreeze proteins.

[Clinical features and carotid artery color Doppler imaging in patients with ocular ischemic syndrome].

OBJECTIVE: To report the clinical manifestations and results of carotid artery color Doppler imaging (CDI) of ocular ischemic syndrome (OIS). METHODS: It was a retrospective case series study. Consecutive patients suspected with OIS were retrospectively studied from 1995 to 2009. We evaluated all patients with medical history, visual acuity, slit lamp, direct and indirect funduscopy, Fundus fluorescence angiography. CDI for internal carotid artery (ICA), Diagnostic Cerebral Angiography and magnetic resonance imaging were also performed. RESULTS: 17 patients (20 eyes) with OIS were studied, 13 males and 4 females. The age range of patients was 46 to 83 years old (median 70). 13 right eyes and 7 left eyes (3 bilateral) were involved. (1) SYMPTOMS: 70.6% (12/17) of the patients complained of visual decrease. 17.6% (3/17) had history of repeated amaurosis fugax (transient monocular visual loss); 11.8% (2/17) patients presented with floaters. The course before the first presentation was 2 days to 6 years (median 3 months). (2) Visual Acuity (VA): among the 20 involved eyes, the best corrected VA < 0.05 in 3 eyes (15.0%); 0.05 to < 0.3 in 7 eyes (35.0%); 0.3 to < 0.7 in 7 eyes (35.0%); ≥ 0.7 in 3 eyes (15.0%). (3) Fundus manifestations: 15.0% (3/20) eyes had cotton-wool spots; 25.0% (5/20) had retinal hemorrhages; retinal arterioles and vein became narrow; venous beading could be seen in severe cases. 30.0% (6/20) had microaneurysms in mid-peripheral area. The pressure of central retinal artery decreased; 25.0% (5/20) had optic disc or retinal neovascularization; 10.0% (2/20) had iris neovascularization. 5.0% (1/20) had neovascular glaucoma; 5.0% (1/20) had optic atrophy. (4) Fundus fluorescence angiography: arm-retinal circulation time prolonged to 14.5 to 39.9 seconds, (median 20.9 seconds). The retinal and choroidal circulation time was also prolonged. The wall of artery and venous could be staining. Hyperfluorescence dots indicating microaneurysms could be seen in the mid-peripheral area. Arterio-venous shunts, non-perfusion area and the leakage of new vessels were observed in some cases. (5) Internal carotid artery CDI examination: The ipsilateral ICA of the 19 involved eyes (another patient with pulseless disease had abnormal subclavian arterial flow) showed completely occluded in 5 cases (26.4%), > 80.0% to 95.0% stenosis in 7 cases (36.8%), 52.0% to 80.0% stenosis in 7 cases (36.8%). Among the 14 contralateral eyes of the same group of patients, CDI showed stenosis of 80.0%, 70.0%, 60.0%, 50.0% and 40.0% in 5 eyes, whereas no fundus abnormal was found. Peak of systolic velocity (PSV): Among the 20 involved eyes of the 17 patients, PSV of the ipsilateral ICA was undetectable in 8 cases which occluded completely or stenosis severely. Among the ipsilateral ICA of the other 12 eyes, 11 cases showed abnormal PSV; PSV increased to 92 to 524 cm/s in 9 cases and decreased to 41 cm/s in 1 case. The blood flow of subclavian artery reduced to 20 cm/s in one patient with pulseless disease. The blood flow of subclavian artery reduced to 20 cm/s in one patient with pulseless disease. CONCLUSIONS: The patients with OIS are usually older than 60 years of age. It is a chronic disease with unnoticed symptoms. The clinical manifestations may include: amaurosis fugax (transient monocular visual loss), hypoperfusion retinopathy and ischemic optic neuropathy. Internal carotid artery severe stenosis or occlusion are the most frequent cause of this disease. CDI is the best safe, reliable and non-invaded method for carotid artery examination.

[Clinical characteristics of choroidal metastasis].

OBJECTIVE: To study the clinical characteristics of choroidal metastasis (CM) to promote the early diagnosis and differentiate from other choroidal tumors. METHODS: Retrospective clinical observational cases. All patients with choroidal metastasis underwent ophthalmologic examination including best corrected visual acuity (VA), slit-lamp examination, binocular indirect funduscopy, color photography, fundus fluorescein angiography (FFA), indocyanine-green angiography (ICGA), optical coherence tomography (OCT), A and B scan ultrasound examination, magnetic resonance image (MRI) as well as CT of the thorax, etc. RESULTS: Nine eyes of 5 patients with CM were examined. Unilateral choroidal involvement was present in 1 patient, bilateral in 4 cases. There were 1 case male and 4 case females. The age of these patients ranged from 31 to 56 years, median 45 years. Ocular symptoms included reduced vision in 4 patients and visual distortion in 1 patient. Visual acuity was 20/400- < 20/63 in four eyes; 20/63- < 20/30 in two eyes and >or= 20/30 in three eyes. The primary cancer site was in the lung in 3 patients, in the breast in 1 patient and in the stomach in 1 patient. Fundus characteristics: Typical CM was more often in the plateau-shaped than in the dome-shaped; yellow-white or mottled in color and associated with subretinal fluid and retinal detachment. The tumor was found in the macular area and juxtapapillary area in 6 eyes, in the area between the macula and the equator in 3 eyes. CM was solitary in 5 eyes and showed multiple lesions in 4 eyes. By FA the lesions showed mottled hyperfluorescence in early stage and leakage in late stage. By ICGA the lesion showed blocked fluorescence and hypofluorescence. Choroidal mass showed moderate irregular internal reflectivity in A-scan ultrasound. B-scan showed a plateau-shaped solid mass. MRI examination of the lesion revealed moderate short T1W and T2W signals. The cancer antigen increased to 16.28 and 4.95 microg/L in two cases. CA125 increased to 160.5 kU/L in one case. CONCLUSIONS: The choroid is the most common site for metastases. CM may precede the diagnosis of primary tumor. Evaluation of A, B scan ultrasound, CT of thorax and cancer antigen test may be important to exclude primary carcinoma from lung and breast in patients with yellow-white in color, plateau-shaped choroidal lesions, especially in both eyes, and without known metastatic diseases.

[The prevalence and risk factors for coronary stenosis in patients with cerebral infarction].

OBJECTIVE: To study the morbidity rate of and relevance to coronary stenosis in cerebral infarction patients. METHODS: CT coronary angiography was performed in 112 cases of cerebral infarction after CT cerebral angiography. Multivariate logistic regression analysis was carried out between the clinical data and coronary stenosis. RESULTS: In 112 cases receiving CT cerebral angiography, the morbidity rate of coronary stenosis was 46.4%. In 95 cerebral infarction patients, the morbidity rate of coronary stenosis was 51.6%. Multivariate logistic analysis showed that age, hypertension, hyperlipidemia, significant narrowing of cerebral artery were identified as independent predictors for coronary stenosis. CONCLUSIONS: Heart examination with 64 row CT should be routinely performed after cerebral angiography in cerebral infarction patients, especially in those with age greater than 65 years, hypertension, hyperlipidemia and significant narrowing of cerebral artery so as to detect coronary stenosis early.

[Clinical manifestations of retinal artery macroaneurysms].

OBJECTIVE: To summarize the clinical characteristics of retinal arterial macroaneurysm and to improve the diagnosis accuracy in order to differentiate it from other fundus diseases. METHODS: It was a retrospective case series. RESULTS: of fluorescein angiography or indocyanine angiography, associated with ultrasound, OCT and several other examinations were analyzed in 22 patients (22 eyes) with retinal arterial macroaneurysm. RESULTS: There were 6 men and 16 women in the 22 patients. The age of all the patients was from 54 to 82 years, and the mean age was 70 years. In the 22 patients, the patient with a single macroaneurysm was 19, and with two macroaneurysms was 3. All 22 patients were affected unilaterally. Macroaneurysms were found only in the temporal artery branches (16 eyes in superior branch and 6 eyes in inferior branch). All macroaneurysms were located in the branches graded 1 to 3. All patients showed pre-retinal hemorrhage and/or vitreous hemorrhage. Sixteen patients suffered from hypertension and five patients had diabetes mellitus. The patient with best corrected visual acuity < 0.05 was 10, 0.05 to 0.2 was 6, 0.3 to 0.6 was 5 and > or = 0.7 was 1. All patients showed retinal arteriosclerosis. After the treatment, 11 patients were pursued,and the patient with best corrected visual acuity < 0.05 was 1, 0.05 to 0.2 was 4 and > or = 0.7 was 1. CONCLUSIONS: Clinical features of retinal arterial macroaneurysm include focal dilation of retinal artery with hemorrhages and exudates, associated with retinal arteriosclerosis. These clinical characteristics can be used to differentiate retinal arterial macroaneurysm from other fundus diseases.

(E)-Methyl 3-{3-[(4-bromo-phen-oxy)-methyl]phen-yl}acrylate.

In the mol-ecule of the title compound, C(17)H(15)BrO(3), the rings make a dihedral angle of 75.54 (17)°. In the crystal structure, inter-molecular C-H⋯O hydrogen bonds link the mol-ecules into centrosymmetric dimers, and the π-stacked dimers inter-act with neighbouring dimers via C-H⋯π stacking inter-actions.

[Discussion of several problems in ophthalmic research in China].

Based on the manuscripts received by several ophthalmologic journals, major issues regarding current ophthalmic research aspects in China were analyzed and discussed, including the selection of project, the design of research, the choice of statistical treatments and the techniques of writing research manuscripts. This article provides suggestions for Chinese ophthalmologists to improve the quality of their research work.

[Measurement of macular edema in retinal vein occlusion using optical coherence tomography and its visual prognosis].

OBJECTIVE: To examine the type of macular edema in patients with retinal vein occlusion by using optical coherence tomography (OCT). To compare the sensitivity and specificity between OCT and fundus fluorescein angiography (FFA). To investigate the visual prognosis and risk factors in patients with cystoid macular edema (CME). METHODS: Ninety-one eyes of 90 patients with various types of retinal vein occlusion were examined by OCT and FFA. There were 54 cases male and 36 cases female. The age of these patients ranged from 20 to 74 years old and averaged (57.8 +/- 13.8) years old. Right eye was affected in 45 cases, and left eye in 46 cases (both eye in 1 case). Central retinal vein, hemicentral retinal vein and branch retinal vein were affected in 54, 9 and 28 eyes, respectively. The average follow-up period was 6.10 months. The sensitivity and specificity of OCT and FFA were measured. The visual prognosis and risk factors were analyzed. Macular central retinal thickness of 54 opposite normal eye was measured for comparison. RESULTS: (1) The classification of macular edema by OCT was cystoid macular edema in 71 eyes (78.0%); subretinal fluid in 14 eyes and laminar macular hole in 1 eye. The minimal and maximal height of intraretinal cystoid space was 94 microm and 1317 microm, respectively and averaged (668.18 +/- 245.58) microm. The minimal and maximal height of macular central retinal thickness of 54 opposite normal eye was 110 microm and 236 microm, respectively and averaged (154.09 +/- 21.85) microm. The maximal height of subretinal fluid space was 1377 microm or even beyond the detective limit of OCT. (2) The sensitivity of OCT for detection of CME was 98.6% and the specificity was 100%. The sensitivity of FFA was 86.1% and the specificity was 100.0%. (3) The visual prognosis of 61 eyes with CME follow-up over 3 months: The difference between the initial and final VA in branch retinal vein occlusion was statistically significant (P < 0.01), while no difference between initial and final VA in central and hemicentral retinal vein occlusion (P > 0.05). The factors affected visual prognosis of CME included the duration, the presence of hemorrhage in cystoid space and the severity of occlusion, etc. In CME under 3 months, the visual prognosis after therapy is better than that of before the therapy (P < 0.01); while in eyes with duration more than 3 months, no difference of vision could be detected before and after the therapy (P > 0.05). CONCLUSIONS: OCT is a safe, high-resolution, non-invasive, reliable and reproducible new examine method for detecting CME. The visual prognosis of CME is poor. It is possible that the visual prognosis can be improved by early detecting CME using OCT.

[Inhibition of lens epithelial cell cycle and PCNA mRNA expression by an antisense oligonucleotides complementary to the messenger RNA of proliferating cell nuclear antigen].

OBJECTIVE: To determine whether antisense oligonucleotides complementary to the messenger RNA of proliferating cell nuclear antigen (PCNA ASODN) inhibit the proliferation of bovine lens epithelial cell (BLEC) by changing the cell cycle and down-regulating the expression of PCNA mRNA and PCNA protein. METHODS: BLECs were cultured in vitro, and the second passage cells were used in this experiment. PCNA ASODN (30 micro mol/L), PCNA SODN (sense oligonucleotides, 30 micro mol/L), basic fibroblast growth factor (bFGF, 10 micro g/L), bFGF (10 ng/ml) + ASODN (30 micro mol/L), bFGF (10 micro g/L) + SODN (30 micro mol/L) were introduced respectively into the medium, and the same amount of PBS was added into the medium as a control. After 24 hours, the cell cycle and the PCNA expression were counted by flow cytometry, and the expression of PCNA mRNA was indicated by Northern ELISA hybridization. RESULTS: PCNA ASODN could decrease the rate of the cells of S phase and down-regulate the expression of PCNA mRNA and PCNA protein. Comparing with the control group, after 24 hours, the rate of cells of S phase was decreased from 15.67% to 7.96%, the expression of PCNA mRNA from 0.266 to 0.176 and the expression of PCNA protein from 55.27% to 12.32%. PCNA ASODN could also inhibit the proliferation of BLEC induced by bFGF, comparing with the bFGF group, the rate of cells of S phase was decreased from 23.4% to 19.9%, the expression of PCNA mRNA from 0.576 to 0.357 and the expression of PCNA protein from 76.4% to 35.48%. CONCLUSIONS: Our results demonstrate that the PCNA ASODN decreases expression of PCNA mRNA and PCNA protein, and stops the cell to enter and progress through S phase. These results provide an important impetus to initiate in vivo studies to determine the feasibility of antisense strategies in the prevention of posterior capsular opacification.

[The occurrence of actual F factor transfer during the stationary-phase mutation in Escherichia coli FC40].

The experiment of adaptive mutation was performed by using Escherichia coli GM133 rif(r) as test cells and HB214 str(r) as scavenger cells. Transfer frequency between GM133 and HB214 was estimated,based on the number of revertants appeared on the selective plates when GM133 were killed by addition of M9 selective medium containing 100 microg/mL of streptomycin at different time. After 30 minutes the cells of GM133 and HB214 were mixed,the estimated transfer frequency was about 0.07%, and two days, 7.47%. After selection of 7 days,some HB214 cells with F' factor from GM133 cells and lac+ mutation were observed, but these cells failed to form the colonies which can be seen by the naked-eye. It was demonstrated that actual F' factor transfer events from test cells GM133 to scavenger cells HB214 occurred during the selection.

[The inhibitory effect of 8-chloroadenosine on tumor necrosis factor-induced proto-oncogene expression of retinal pigment epithelial cells].

OBJECTIVE: To investigate the inhibitory effect of 8-chloroadenodine (8-CLA) on tumor necrosis factor-alpha (TNF-alpha)-induced proto-oncogene expression in retinal pigment epithelial (RPE) cells. METHODS: Primary culture and subculture of health bovine RPE cells were established in vitro. RPE cells were treated with tumor necrosis factor TNF-alpha (13.3 mmol/L) with or without 8-CLA (16 micro mol/L) for 30 minutes or 3 hours, respectively. C-fos and c-myc mRNA expression in RPE cells was detected by Northern blot analysis. RESULTS: Normal RPE cells expressed c-fos and c-myc mRNA at a low level. After treatment with TNF-alpha, the expression of c-fos and c-myc mRNA was increased by 3.13 and 1.5 fold, respectively, as compared to untreated cells. 8-CLA decreased TNF-alpha-induced c-fos and c-myc mRNA expression by 25.0% and 29.0%, respectively. CONCLUSIONS: 8-CLA can inhibit TNF-alpha-induced c-fos and c-myc mRNA expression in RPE cells. These findings may be useful for exploring new drugs for inhibiting the growth-induced proliferation of RPE.