BackgroundRetinal pigment epithelial(RPE) cells can secrete platelet-derived growth factor (PDGF) and PDGF receptor(PDGFR).Studies have shown that PDGF plays a key role in the formation of proliferative vitreous retinopathy(PVR). ObjectiveThis study was to investigate the proliferation and apoptosis changes of RPE after blockage of the PDGFR-α expression by antisense oligonucleotide ( ASODN ) in vitro. Methods Human RPE cells strain was cultured in low glucose DMEM with 10％ fetal bovine serum.Logarithmic phase cells were collected and incubated in 96-well plate at the density of 5 × 105 cells/hole.PDGFR-α ASODN was transfected into RPE cells at different concentrations for 48 hours.The cells of the blank control group were regularly cultured without any transfection.The changes of PDGFR-α expression were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the proliferation of RPE was detected by MTT as the A490 value.Hoechst 33258 fluorescence staining was used to determine the apoptosis of RPE.Flow cytometry method (FCM) was applied to detect the change of cell cycle and apoptosis rate of RPE cells. ResultsThe A490 values of RPE cells were 1.45±0.12,1.07±0.06,0.65±0.05 in blank control group,1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group with the significant difference(P=0.00 ),and that of 1.0 μmol/L Lipo-ASODN group and 2.0 μ mol/L Lipo-ASODN group were significantly lower than the blank control group ( P =0.00,0.00).Hoechst 33258 staining showed that the apoptosis cells were obviously more in Lipo-ASODN group compared with blank control group.PDGFR-α ASODN transfection induced an increase of percentage of RPE cells in G0/G1 phase( F =206.70,P =0.00),and the apoptosis rates in 1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group were significantly enhanced in comparison with blank control group ( 37.8 ± 1.3 vs 10.5 ± 0.1,61.2 ± 1.9 vs 10.5 ± 0.1 ) ( F =1808.90,P =0.00 ).Expression intensity of PDGFR-α mRNA in RPE cells in Lipo-ASODN groups was lower. ConclusionsBlocking the PDGFR-α expression with ASODN technology can suppress proliferation and induce apoptosis of RPE cells.Intensity of PDGFR-α mRNA expression in RPE cells is ASODN dose-dependent.ASODN targeted to PDGFR-α offers an experimental basis of the gene therapy for PVR.

Objective　To investigate the mechanism affecting on permeability of vascular endothelial cell by nitric oxide (NO). Methods　Series concentration of sin-1（a donor of NO） were added to ECV 304, a cell line of human umbilical vein endothelium. Cell growth and expression of f-actin, a cytoskeleton protein were observed. Results　Cell growth was inhibited with a dose from 6.25 to 100 μmol/L and was caused to death at the concentration of 50 to 100 μmol/L by sin-1. The expression of f-actin was suppressed obviously after cultured with 100 μmol/L sin-1 for 4 hours. Conclusion　It suggests that anomaly increased NO can increase permeability of blood vessels by suppressing the expression of f-actin, inhibiting cell growth or even resulting in cell death.

AIM: To explore the nursing cooperation highlights of eight osteopetrosis patients underwent optic nerve decompression via transsphenoidal microsurgical approach instead of routine pathway, and to improve the quality of nursing cooperation. ·METHODS: We enrolled 8 cases ( left eye in 3 cases, right eye in 5 cases ) of osteopetrosis patients referred from the Eye Hospital of Wenzhou Medical University during February 2012 to November 2016. Patients received ophthalmic examinations including visual acuity and diagnostic imaging tests in pre-operation and post -operation. All eyes were performed surgical optic nerve decompression through endoscopic approach in assist of image guidance system. We retrospectively analyzed the clinical data and surgical cooperation procedure of these cases and summarized nursing cooperation experience. ·RESULTS:The operations of 8 patients were completed successfully without massive hemorrhage. Mean visual acuity improved from pre-operation (2. 5±2. 1) to post-operation (3. 4±1. 9). Cerebrospinal fluid leakage occurred in 1 patient and was instantly repaired during the operation. We performed the nursing strategy as postural drainage, condition monitoring and conscious assessment intra-and post-operation. ·CONCLUSION: It is the critical for this kind of surgery that both circulating nurse's high-skilled cooperation to the connection and operation of the navigation system, to treat with complication during the surgery, and scrub nurse's sufficient preparation of surgical instruments and consumables, proficient equipment delivery, meticulous management, use and maintenance of equipment.

OBJECTIVETo evaluate the efficacy and safety of fast track surgery(FTS) in patients undergoing laparoscopic colorectal resection.

METHODRandomized controlled trials(RCT) or clinical controlled trials(CCT) on fast-track surgery in patients undergoing laparoscopic colorectal resection were obtained from databases including CNKI, Wanfang, PubMed, EMBACE, and Cochrane Library between January 2000 and March 2012. Meta-analysis was performed with RevMan 5.1.

RESULTThere were 6 RCTs and 7 CCTs including 1795 patients. There were 955 patients in the FTS group and 840 in the control group. The time to passage of flatus was shorter(WMD=-1.37, 95%CI:-1.55~-1.19, P<0.05), time to resumption of diet/drink was shorter(WMD=-2.62, 95%CI:-2.69~-2.55, P<0.05), length of postoperative hospital stay was decreased(WMD=-1.63, 95%CI:-1.92~-1.34, P<0.05) and the incidence of postoperative complications were less(OR=0.52, 95%CI:0.41~0.67, P<0.05) in the FTS group. However, there were no differences in readmission(P>0.05).

CONCLUSIONFast-track surgery in patients undergoing laparoscopic colorectal resection can promote bowel function recovery, decrease the incidence of postoperative complications and length of hospital stay.

Controlled Clinical Trials as Topic ; Digestive System Surgical Procedures ; Humans ; Incidence ; Laparoscopy ; Length of Stay ; Postoperative Complications

Objective: To analyze the association between different sleep behaviors and overweight and obesity of junior high school students in Yangzhou City, and to provide a basis for policies and interventions related to adolescent health management. Methods: A total of 1 589 students in grades 7-9 from two middle schools in Yangzhou City were selected using the cluster sampling method and were administered with sleep time, bedtime, social jetlag difference, and sleep habits. Results: Totally 64.38% were sleep deprived during the school days, 86.78% went to bed too late, 46.51% had a social jetlag of ≥1 h, and 37.44% took a nap every day(Incluldes holidays and school days). Social jetlag length was statistically different between grades( F =6.97, P < 0.01 ). Girls[(0.95±0.65)h] shown significantly higher social jetlag than the boys[(0.76±0.59)h]( t=6.19, P <0.01). Later bedtime on weekends, later wake up time on weekends and poor sleep behavior were risk factors for overweight and obesity in junior high school students( OR=1.20, 1.14, 1.04, P <0.05). Conclusion Junior high school students had less sleep and later bedtimes with the increase of grade, and weekend bedtimes,wake up times and poor sleep behavior were independently associated with the risk of overweight and obesity in junior high school students. Parents and schools should be instructed to pay attention to their sleep health and carry out adolescent sleep health guidance.

AIMTo study the anti-tumor bioactive steroid saponins of Paris vietnamensis (Takht.).

METHODSThe constituents were isolated and purified by chromatography and their structures were identified by spectral analysis and physicochemical properties. The cytotoxic bioactivities of the constituents were determined by MTT.

RESULTSEleven compounds were obtained and identified as 3beta, 5alpha,6alpha-trihydroxy-7(8)-en-isospirostanol-3-O-beta-D-glucopyranosyl(1-->3) [alpha-L-rhamnopyranosyl(1-->2)]-beta-D-glucopyranoside (1), which was named as parisvietnaside A, 25 (R) diosgenin-3-O-alpha-L-arabinofuranosyl(1-->4)-beta-D-glucopyranoside (2), 25(R) diosgenin-3-O-alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucopyranoside (3), 25 (R) diosgenin-3-O-alpha-L-arabinofuranosyl (1-->4) [alpha-L-rhamnopyranosyl (1-->2)]-beta-D-glucopyranoside (4), 25 (R) diosgenin-3-O-beta-D-glucopyranosyl (1-->3) [alpha-L-rhamnopyranosyl (1-->2)]-beta-D-glucopyranoside (5), 25 (R) diosgenin-3-O-alpha-L-rhamnopyranosyl (1-->4)-alpha-L-rhamnopyranosyl(1-->4) [alpha-L-rhamnopyranosyl-(1-->2)]-beta-D-glucopyranoside (6), 25 (R) pennogenin-3-O-alpha-L-arabinofuranosyl(1-->4)-beta-D-glucopyranoside (7), 25 (R) pennogenin-3-O-alpha-L-rhamnopyranosyl (1-->2)-beta-D-glucopyranoside (8), 25 (R) pennogenin-3-O-alpha-L-arabinofuranosyl (1-->4) [alpha-L-rhamnopyranosyl-(1-->2)]-beta-D-glucopyranoside (9), 25 (R) pennogenin-3-O-beta-D-glycopyranosyl (1-->3) [alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucopyranoside (10) and 25 (R) pennogenin-3-O-alpha-L-rhamnopyranosyl (1-->4)-alpha-L-rhamnopyranosyl(1-->4) [alpha-L-rhamnopyranosyl-(1-->2)]-beta-D-glucopyranoside (11). Some constituents had cytotoxic bioactivities.

CONCLUSIONCompound 1 is a new spirostanol saponin, and compounds 2, 3, 6-11 were obtained from Paris vietnamensis (Takht.) for the first time. Compounds 3, 4, 6, 8 had cytotoxic bioactivities against tumor cells HepG2 and SGC-7901.

Adenocarcinoma ; pathology ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Liliaceae ; chemistry ; Liver Neoplasms ; pathology ; Molecular Conformation ; Molecular Structure ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Saponins ; chemistry ; isolation & purification ; pharmacology ; Stomach Neoplasms ; pathology

Animals ; China ; epidemiology ; Female ; Humans ; Incidence ; Male ; Schistosoma japonicum ; Schistosomiasis japonica ; epidemiology ; transmission ; Urban Health

Objective: To establish a solvent desorption gas chromatographic method for determination of Sevoflurane, Isoflurane and Enflurane in the air of the Workplace. Methods: Sevoflurane, Isoflurane and Enflurane were collected with activated carbon tube and desorbed with dichloromethane, separated with DB-1 capillary columns, and then detected with flame ionization detector. Results: The linearity ranges were 1.9-304.8 μg/ml for Sevoflurane, 2.1-300.4 μg/ml for Isoflurane and 1.7-305.2 μg/ml for Enflurane, The correlation coefficient was both >0.999. Their limits of detection were 0.6 μg/ml, 0.6 μg/ml and 0.5 μg/ml, and Their limits of quatification were 1.9 μg/ml, 2.1 μg/ml and 1.7 μg/ml, and their minimum detectable concentrations were 0.1、0.2 and 0.1 mg/m³ per 4.5 L of air. Their relative standard deviations (RSD) were 2.5%-3.0%, 2.3%-3.1% and 2.2%-3.0%. The average desorption efficiencies were 101.1%-103.3%, 100.7%-102.7% and 101.0%-102.9%. The sampling efficiency was both 100%. The breakthrough volume of 100 mg actived carbon was 3.7 mg, 3.4 mg and 3.4 mg. Sevoflurane, Isoflurane and Enflurane in activated carbon tube could be kept at least 10 days at room temperature without significant losses. Conclusion The method shows lower detection limit, high accuracy and precision. It is feasible for determination of Sevoflurane, Isoflurane and Enflurane in the air of workplace.

A bacterial strain DN25, effective on cyanide-degradation, was isolated from contaminated soil and identified as Alcaligenes sp. on the basis of phenotype analysis and 16S rDNA sequence analysis. It showed great tolerance to the cyanide, which can grow in the medium containing 500mg CN -/L. The suitable condition for the cell growth and boitransformation was pH8.0 and 30oC and the transformation rate for 500mg CN - /L could achieve 99% in 10 h. It has also been found that the screened strain had the ability of K 4Fe(CN) 6 transformation with 96% of transformation rate at 12 h for the concentration of 500 mg CN /L.

In order to confirm the role that the 49th amino acid residue plays in enzymatic inactivity of Glutamine 49 phospholipase A2(Gln49-PLA2),site-directed mutagenesis of its 49th amino acid gene codon was conducted using PCR.Aspartic acid 49 phospholipase A2(Asp49-PLA2-Q49D-PLA2),the mutant of Gln49-PLA2 was expressed in E.coli with pET32a+ vector.The fusion protein,expressed as inclusion body,after being denatured,was on-column refolded and purified by immobilized metal affinity chromatography(IMAC),and then cleaved by Factor Xa.The mature Q49D-PLA2 mutant was obtained by Hitrap SP cation exchange and Superdex 75 gel filtration chromatography,with the recovery rate of 1.3%,and the specific activity of the mature Q49D-PLA2 mutant was 72 U/mg.It has been demonstrated that the 49th glutamine amino acid residue is the main reason in enzymatic inactivity of Gln49-PLA2 and the results are helpful for denatured protein refolding,especially in rich disulfide bonds conditions.