Objective To analyze the correlation of visual acuity and metamorphopsia with foveal morphology before and after vitrectomy with internal limiting membrane peeling (ILMP) in idiopathic macular epiretinal membrane (IMEM) eyes.Methods This is a retrospective case series of 47 IMEM patients (49 eyes).All the patients underwent 25G pars plana vitrectomy with IMEM removal and ILMP.The best corrected visual acuity (BCVA) was measured using the international standard visual acuity chart,and the results were converted to the logarithm of the minimum angle of resolution (logMAR) visual acuity.The severity of metamorphopsia was measured using M-charts.The central macular thickness (CMT),inner nuclear layer thickness (INT),inner retinal layer thickness (IRT),outer retinal layer thickness (ORT),the status of ellipsoid zone (EZ) were assessed with spectral-domain optical coherence tomography at baseline and each month postoperatively.The differences in BCVA,CMT,INT,IRT,ORT and status of EZ before and after surgery were analyzed,so did the correlations between these indexes at the same time.Results Compared with baseline,the postoperative BCVA was significantly increased (F=6.133,P＜0.001),but the M value,CMT,INT,IRT,ORT were significantly decreased (F=12.481,10.565,15.739,6.046,10.569;P＜0.001);the integrity of EZ was improved significantly (x2=12.309,P＜0.001).Preoperative BCVA was positively related to the CMT (r=0.720) and ORT (r=0.720,0.887;P＜0.05),while negatively related to preoperative integrity of EZ (r=-0.295,P＜0.05).The postoperative BCVA was positively related to the preoperative BCVA and postoperative CMT,ORT (r=0.774,0.754,0.842;P＜0.05),while negatively related to postoperative integrity of EZ (r=-0.676,P＜0.05).The preoperative M value was positively related to the preoperative CMT,INT,IRT,and ORT (r=0.931,0.668,0.840,0.637;P＜ 0.05).The postoperative M value was positively related to the preoperative M value and postoperative CMT,INT,IRT,and ORT (r=0.723,0.722,0.767,0.825,0.387;P＜0.05).Conclusions Vitrectomy with ILMP for IMEM can improve the visual acuity,metamorphopsia and foveal morphology.Both visual acuity and metamorphopsia correlate with foveal morphology.

Objective To evaluate the safety and effectiveness of vitrectomy combined with internal limiting membrane (ILM) tamping on macular hole and retinal detachment (MHRD) in highly myopic eyes.Methods 23 patients (23 eyes) were retrospectively reviewed,who were diagnosed as MHRD through examination of the ocular ftmdus,optic coherence tomography (OCT) and B-mode ultrasonography.There were 5 males (5 eyes) and 18 females (18 eyes).The mean age was (62.35 ± 8.28) years.The mean course of disease was 1.1 months.The logarithm of the minimum angle of resolution (logMAR) best-corrected visual acuity (BCVA) was 2.31 ± 0.72.The mean axial length was (28.66 ± 1.99) mm.All patients underwent 23G microincision vitrectomy.After vitreous gel and cortex were gently resected,the ILM around the edges of the macular hole was stained with indocyanine green,and was folded and pushed to fill the macular hole gently.Then silicone oil or C3F8 gas tamponade was applied in 18 eyes and 5 eyes,respectively.The silicone oil was removed after 3 months.The follow-up was 6 months.The BCVA,macular hole closure,retinal anatomical reattachment were retrospectively observed,and were used to evaluate the safety and effectiveness of the surgery.Results At the 6 months after surgery,the logMAR BCVA was improved to 1.13 ± 0.38,the difference was significant (t=l 5.33,P=0.00).The postoperative macular hole closure rate and retinal anatomical reattachment rate were 100％.There were no ocular or systemic adverse events observed in all patients.Conclusion Vitrectomy combined with ILM tamping is an effective and safe treatment for the high myopic eyes with MHRD.

Objective To investigate the effects of microRNA-21-5p (miR-21-5p) on hyperoxic acute lung injury (HALI) in rats and provide a theoretical basis for HALI gene therapy. Methods One hundred and sixty Sprague-Dawley (SD) rats were randomly divided into four groups with number table:hyperoxia control group, phosphate buffer saline (PBS) group, blank virus group and miRNA-21-5p group (each, n = 40). The rats in hyperoxia control group were fed directly in the hyperoxia box (oxygen concentration > 90%); in the other three groups, 200 μL PBS, 200μL slow virus and 200μL miRNA-21-5p slow virus were dropped into the nose respectively, and then they were fed in the hyperoxia box. The rats were exposed to hyperoxia in the boxes for 0, 24, 48 and 72 hours in all the groups, and at each time point, 10 rats were taken randomly from each group to perform arterial blood-gas analysis, calculate oxygenation index (OI) and respiratory index (RI). Afterwards the rats were sacrificed by blood-letting from carotid artery under intra-peritoneal anesthesia, and the lung tissues were obtained to measure the left lung wet/dry weight (W/D) ratio, hemotoxylin-eosin (HE) staining was made and the pathological changes of the right lung were observed under light microscope and the pathological score was measured. Results At 0 hour, the OI, RI, lung W/D ratio and the lung tissue pathology score in rats with hyperoxic injury had no statistically significant differences among the four groups (all P>0.05). With the extension of time, the level of OI was gradually reduced, and the levels of RI, pathologic score and W/D ratio of lung tissues were gradually increased. Compared with the hyperoxia control group, in miRNA-21-5p group, the levels of OI were increased significantly at 24, 48 and 72 hours after the exposure to hyperoxia [mmHg (1 mmHg = 0.133 kPa): 24 hours 358.10±29.25 vs. 306.19±37.23, 48 hours 336.67±29.27 vs. 269.70±29.00, 72 hours 323.81±19.05 vs. 203.81±43.40, all P < 0.05], whereas the levels of RI were decreased significantly (24 hours 0.23±0.05 vs. 0.31±0.06, 48 hours 0.28±0.07 vs. 0.38±0.06, 72 hours 0.30±0.04 vs. 0.46±0.07, all P <0.05), the pathologic scores were decreased significantly (24 hours 0.60±0.52 vs. 0.90±0.74, 48 hours 1.30±0.95 vs.2.90±1.20, 72 hours 1.90±0.88 vs. 4.70±1.57, all P < 0.05) and the levels of W/D ratio were decreased obviously (24 hours 3.77±0.38 vs. 4.14±0.46, 48 hours 3.83±0.31 vs. 4.56±0.34, 72 hours 3.89±0.31 vs. 5.32±0.27, all P<0.05). Compared with the hyperoxia control group, the index results of the PBS group and the blank virus group after staying in the box had no statistical significant differences at each time point (all P>0.05). Under the optical microscope, along with the prolongation of exposure to hyperoxia, the structure of alveoli was gradually disturbed, their walls fractured and damaged, alveolar septa widened, edematous, infiltrated with inflammatory cells and in part of the rats a small amount of red blood cell exudates could be seen, but the degree of lung pathological injury in miRNA-21-5p group was much milder than that of the other groups. Conclusion The rat persistently exposed to hyperoxia for 24 hours can establish the rat model of HALI successfully, and the miRNA-21-5p can protect the lung tissue from the damage to some degrees in HALI rats.

Ammonium perchlorate (AP), mainly used as solid propellants, was reported to interfere with homeostasis via competitive inhibition of iodide uptake. However, detailed mechanisms remain to be elucidated. In this study, AP was administered at 0, 130, 260 and 520 mg/kg every day to 24 male SD rats for 13 weeks. The concentrations of iodine in urine, serum thyroid hormones levels, total iodine, relative iodine and total protein, and malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) activity in thyroid tissues were measured, respectively. Our results showed that high-dose perchlorate induced a significant increase in urinary iodine and serum thyroid stimulating hormone (TSH), with a decrease of total iodine and relative iodine content. Meanwhile, free thyroxine (FT4) was decreased and CAT activity was remarkably increased. Particularly, the CAT activity was increased in a dose-dependent manner. These results suggested that CAT might be enhanced to promote the synthesis of iodine, resulting in elevated urinary iodine level. Furthermore, these findings suggested that iodine in the urine and CAT activity in the thyroid might be used as biomarkers for exposure to AP, associated with thyroid hormone indicators such as TSH, FT4.

ObjectiveTo study the effect of hydrogen peroxide (H2O2) in inducing apoptosis of typeⅡalveolar epithelial cell (AECⅡ) after overexpression by adenoviral transfection of micro RNA-21-5p (miR-21-5p), and to explore the mechanism of its anti-apoptosis.Methods Subculture AECⅡ were randomly divided into four groups: normal control group (normal saline), H2O2 challenge group ( 0.5 mmol/L H2O2), miR-21-5p overexpression group (miR-21-5p adenovirus+ 0.5 mmol/L H2O2), miR-21-5p negative transfection group (adenovirus void+0.5 mmol/L H2O2). Transmission electron microscopy and flow cytometry were used to detect apoptotic morphology and early apoptotic rate. Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of miR-21-5p in AECⅡ, and Western Blot was used to detect the protein expressions of Bcl-2, Bax, and caspase-3 at the highest transfection efficiency at different time points (6, 12, 24, 48 hours).Results ① AECⅡ identification: fluorescence microscopy showed the presence of characteristic structure of AECⅡ, i.e. microvilli and osmiophilic lamellar bodies.② Apoptotic morphology: transmission electron microscopy showed cytoplasmic retraction, chromatin condensation, margination, lack of cell surface microvilli, and emptying of osmiophilic lamellar bodies in AECⅡ.③ The expression of miR-21-5p in AECⅡ: the highest transfection efficiency was found at 48 hours. The expression of miR-21-5p in miR-21-5p overexpression group was significantly higher than that of the normal control group, H2O2 challenge group and miR-21-5p negative transfection group (A value: 1.54±0.02 vs. 1.02±0.02, 0.56±0.03, 0.58±0.02, allP< 0.05).④ The rate of early apoptosis: compared with normal control group, the early apoptotic rates in H2O2 challenge group, miR-21-5p negative transfection group and miR-21-5p overexpression group were gradually elevated with the prolongation of injury time. The early apoptotic rate in miR-21-5p overexpression group was significantly lower than that of the H2O2 challenge group and miR-21-5p negative transfection group at all time points except 6 hours [12 hours: (10.73±2.80)% vs. (16.26±0.59)%, (16.04±0.70)%; 24 hours:(16.00±3.44)% vs. (23.29±2.78)%, (23.58±2.31)%; 48 hours: (31.30±3.55)% vs. (50.53±2.17)%, (49.41±1.97)%, allP< 0.05]. There was no significant difference in early apoptotic rate between miR-21-5p negative transfection group and H2O2 challenge group at each time point.⑤ Protein expression: the expressions of Bax and caspase-3 in miR-21-5p overexpression group were significantly lower than those of the H2O2 challenge group and miR-21-5p negative transfection group [Bax (A value): 0.07±0.01 vs. 0.18±0.01, 0.13±0.01; caspase-3 (A value): 0.07±0.01 vs. 0.23±0.01, 0.12±0.01, allP< 0.05], and Bcl-2 protein expression was significantly higher than that of the H2O2 challenge group and miR-21-5p negative transfection group (A value: 0.26±0.01 vs. 0.06±0.01, 0.10±0.01, both P< 0.05).Conclusions① miR-21-5p has the function of anti-apoptosis of AECⅡ.② Adenoviral vector is a successful gene transfer vector when transfected with AECⅡ.③ The anti-apoptosis of AECⅡ by miR-21-5p may be associated with reduced Bax and caspase-3 protein levels and raised expression levels of Bcl-2 protein.

Objective To explore a simpler, more economic and reproducible method to reproduce a model of high oxygen induced acute lung injury (HALI) in rats. Methods An animal feeding box equipped with a controllable high oxygen was designed. 100 Sprague-Dawley (SD) rats were divided into normal control group and HALI group by random number table method, with 50 rats in each group. Each group was randomly subdivided into five subgroups according to the duration of exposure to high oxygen, namely 0, 24, 48, 72 and 96-hour subgroups, with 10 rats in each subgroup. The rats in normal control group were kept in cages with ambient air, and the rats in HALI group were kept in an oxygen tank in which the oxygen concentration was higher than 90% volume ratio, with the temperature maintained at 25-27 ℃, humidity of 50%-70%, and CO2 concentration < 0.5% for 23.5 hours every day. The arterial blood of rats was collected for analysis of blood gas at all time points, and the oxygenation index (OI) and respiratory index (RI) were calculated. Then the rats were sacrificed and the right lung was harvested, which was sectioned and stained with hematoxylin and eosin (HE). The changes in histopathology were observed with light microscopy, and pathological score was recorded. The left lung was harvested for the measurement of the wet/dry weight ratio (W/D). Results With the prolongation of high oxygen exposure time, the degree of lung injury in HALI group was gradually increased, and the degree of derangement of alveolar structure appeared in an increasing degree, with destruction of the alveolar wall, widening of alveolar space, and appearance of edema, and inflammatory cell infiltration. A small quantity of red blood cells exudation could be found in some rats. The pathologic changes were most obvious at 48-72 hours after exposure. With the prolongation of high oxygen exposure time (0, 24, 48, 72, 96 hours), the OI (mmHg, 1 mmHg = 0.133 kPa) in HALI group was gradually decreased (446.67±29.93, 306.19±37.23, 269.70±29.00, 253.81±43.40 and 245.58±35.25), RI, pathological score of lung tissue and W/D ratio were gradually increased [RI: 0.25±0.04, 0.31±0.06, 0.38±0.06, 0.46±0.07 and 0.44±0.03; pathological score of lung tissue: 0.00±0.00, 0.90±0.74, 2.90±1.20, 4.70±1.57 and 4.80±1.23; lung W/D ratio: 3.84±0.61, 4.14±0.46, 4.56±0.34, 5.32±0.27 and 5.18±0.25]. Statistically significant differences were found in 72-hour group as compared with that of other groups (all P < 0.05), while no significant difference was found between 96 hours and 72 hours groups (all P > 0.05). There were significant differences in changes between 24, 48, 72, and 96 hours as compared with those of the normal control group: OI (mmHg): 24 h 306.19±37.23 vs. 435.65±25.34 and 96 h 245.58±35.25 vs. 465.42±24.75; RI: 24 h 0.31±0.06 vs. 0.24±0.04 and 96 h 0.44±0.03 vs. 0.24±0.06. The same as true in pathological scores of lung tissue: 24 h 0.90±0.74 vs. 0.00±0.00 and 96 h 4.80±1.23 vs. 0.00±0.00; lung W/D ratio: 24 h 4.14±0.46 vs. 3.79±0.44 and 96 h 5.18±0.25 vs. 4.12±0.91, all P < 0.05. Conclusions A self-designed high oxygen box is simple, easy to operate and reproduction of HALI model can be attained. Sustained exposure to high concentrations of oxygen (≥ 90%) for 24 hours can replicate the HALI model successfully, and the most serious injury appears at 48-72 hours after exposure.

Hepatitis B surface antigen (HBsAg) carrying preS sequences could be an ideal candidate for a new hepatitis B virus (HBV) vaccine with higher efficacy. Here we report the success in achieving efficient and stable expression of hepatitis B virus S antigen and preS1 epitope fusion protein (S/preS1) in CHO cells. The HMRCHEF53u/Neo-S/preS1 expression vector carrying S/preS1 gene was constructed and transfected into CHO-S cells. A stable and high-expression CHO cell line, named 10G6, was selected by ELISA and limiting dilution analysis. Western blotting analysis showed S/preS1 expressed from 10G6 cells possessed both S and preS1 antigenicity. 10G6 cells displayed characters of favorable growth and stable S/preS1 expression in repeated batch cultures as evaluated by viable cell density, viability and S/preS1 concentration. And cultivation of 10G6 cells in fed-batch mode resulted in S/preS1 production at 17-20 mg/L with viable cell density at 7 x 10(6)-10 x 10(6) cells/mL.
Animals ; CHO Cells ; Cricetulus ; Epitopes ; biosynthesis ; genetics ; Hepatitis B Surface Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B Vaccines ; biosynthesis ; genetics ; Hepatitis B virus ; Protein Precursors ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

HepG2 ceils were treated with various concentrations of tumor necrosis factor-or (TNF-α) for 24 hours. RT-PCR and Western blot were used to measure protein tyrosine phosphatase-1 B(PTP-1B)expression, and luciferase reporter assay was used to detect NF-kB activity. The results showed that treatment of HepG2 cells with TNF-α for 24 hours led to upregulation of PTP-1B and NF-kB activity in a dose-dependent manner. Inhibition of NF-kB by PI)TC significantly attenuated TNF-α-induced PTP-IB expression in HepG2 cells. Thus, the transactivation of NF-kB seems to play an important role in the expression of PTP-1B in HepG2 cells induced by TNF-α.

OBJECTIVE: To explore the relationship between polymorphism of apolipoprotein E (ApoE) exon 4 gene and different syndromes in patients with coronary heart disease (CHD). METHODS: Two hundred patients with CHD were divided into four groups according to syndrome differentiation, including syndrome of phlegm (PS), syndrome of blood stasis (BSS), syndrome of phlegm-blood stasis blocking (PBBS) and syndrome of non-phlegm and non-blood stasis (NPNBS). One hundred healthy volunteers were included in control group. Blood lipids were measured by routine examination. Total DNA of peripheral blood was extracted. ApoE genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. All data were analyzed by SAS software. RESULTS: (1)The occurrence rate of epsilon4 allele of ApoE in patients with CHD was 19.5%, significantly higher than 9.5% in the control group (P<0.05), and the E 3/4 genotype was especially more frequent (P<0.01). (2) The levels of total cholesterol (TC), total triglycerides (TG) and low-density lipoprotein cholesterol (LDL-C) in patients with epsilon4 were higher than those in patients without epsilon4 (P<0.01). (3) The frequencies of epsilon4 allele and E3/4 genotype in patients with PS were significantly higher than those in patients with BSS (P<0.05). CONCLUSION: ApoE epsilon4 allele, especially E3/4 genotype, is the risk factor of CHD. There is a relatively close relationship between patients with ApoE epsilon4 allele and PS. It may be one of the main susceptible genes in CHD patients with PS.

Objective: To study the therapeutic effects of combined treatment with Chinese and western medicine in children with cerebral palsy (CP). Methods: 104 CP children were treated mainly with physiotherapy and acupuncture, together with occupational therapy, sensory integration and electrical stimulation. Gross Motor Function Measure (GMFM) was used to assess the effects of treatment. Results: After treatment for 3.29±0.80 months, the majority of CP children improved their IQ and limb symptoms, with decreased abnormal posture, increased muscle strength and improved motor developmental function. GMFM88 and GMFM66 scores had significantly increased after treatment. Conclusions: Combined treatment with Chinese and Western Medicine has confirmed effects on CP children.