Background Studies demonstrated that human amniotic epithelial cells (AECs) have some characteristics of embryonic stem cells and they were used to re-establish the surface of eyes. Human AECs may serve as new seed cells in tissue engineering for corneal epithelium reconstitution in the future. Objective The present study was to investigate the application value of human amniotic epithelium cells transfected by lentiviral vectormediated enhanced green fluorescent protein (EGFP) gene as new seed cell source for engineering the corneal surfacelayer. Methods Lentiviral vector carrying the objective gene EGFP was transfected into human amniotic epithelial cells (pLenti6/V5-DEST),and the transient expression of the transgene in the human amniotic epithelial cells was observed under the fluorescence microscope. Flow cytometry was used to detect the positive expression rates of EGFP in transfected cells. The transfected human amniotic epithelial cells were seeded onto the fresh corneal stromal surface of New Zealand white rabbit and cultured in vitro. The stem cell deficiency ( SCD ) models were established by cutting off the limbus of cornea in 20 eyes of New Zealand white rabbits, and the model rabbits were then divided into 2 groups randomly. The transplanted grafts carrying the pLenti6/V5-DEST-EGFP gene-transferred human amniotic epithelium cells were regarded as the pLenti6/V5-DEST-EGFP group, and the corneal stroma graft without any epithelial cell served as the control group. The opacity of stroma and corneal conjunctivalization and vascularization were observed daily. The rabbits' eyes were extracted one month after operation. The expression of EGFP in the cornea was detected under the fluorescence microscope, and the expression of CK8, CK18 and CK12 in cornea was detected by immunohistochemical staining. Results The shape of the transferred human amniotic epithelial cells resembled normal human amniotic epithelial cells. 48 hours after the transient transfection of EGFP presented with the highest expression level throughout the observation duration, with a positive expression rate of EGFP of 61.5% ,showing significant differences in comparison with that of 12 ( 5.24% ) , 24 ( 38.27% ) or 96 ( 39. 10% ) hours ( P ＜0. 05) post-transfection; but no obvious difference was found in the positive rate of transiently transfected EGFP between 48 hours and 72 hours ( 58.36% ) ( P＞0. 05 ). Six cornea grafts were clear in 1 month and two corneas were rejected during the observation period in the pLenti6/V5-DEST-EGFP group. A few new blood vessels were seen around the graft. Ten corneas of the control group became opaque and cloudy with new blood vessels growth around the grafts. Imunohistochemistry revealed the positive expressions of CK8, CK1 8 and CK12 in the corneal epithelial layer in the pLenti6/V5-DEST-EGFP group. However,the expression of CK12 was absent in the control group. Conclusion Human amniotic epithelium cells transfected with the pLenti6/V5-DEST-EGFP gene is a new and ideal feed cell type to reconstruct the corneal surface layer. Lentivirus is a relatively safe gene transfection vector.

OBJECTIVETo determine the mechanism of pseudo-tears of the lateral meniscus caused by the transverse geniculate ligament (TL) and the miniscofemoral ligament(MFL) and to investigate a method to differentiate pseudo-tears from true tear of the lateral meniscus.

METHODSForm June 2012 to February 2014, MR examinations of 72 knees (44 left knees and 28 right knees) without tear of the lateral meniscus verified by arthroscopy were performed in the sagittal and coronal plane. There were 41 males and 31 females in the group, with an average age of 33.7 years old (ranged from 25 to 61). The MR appearance of the TL and the MFL was carefully observed.

RESULTSThere existed fatty tissue in the gap between the TL and the anterior horn of the lateral meniscus and its central tendinous attachment. On the sagittal images, the fatty tissue formed a linear high-signal cleft between the TL and the anterior horn of the lateral meniscus. This might be mistaken as an oblique tear within the anterior horn of the lateral meniscus. It was called as pseudo-tears of the anterior horn of the lateral meniscus. In sagittal plane, the MFL was identified as a circle-like or short stick-like area of low signal intensity anterior or posterior to the posterior cruciateligament. Nevertheless, a belt-shaped area of low signal intensity from the posterior horn of the lateral meniscus to lateral facet of the medial femoral condyle was identified in the coronal plane. A linear area of high signal intensity between the MFL and the lateral meniscus was found in sagittal plane, which might be mistaken as an oblique tear within the posterior horn of the lateral meniscus. It was called pseudo-tears of the posterior horn of the lateral meniscus. The occurrence rate of the TL was 34.7% (25/72). The prevaleribe of pseudo-tears of the anterior horn of the lateral meniscus was 18 cases. The shape of the anterior horn of the lateral meniscus was regular, and the course of the pseudo-tears cleft was oblique. The occurrence rate of the MFL was 73.6% (53/72), which included the anterior MFL 23.6% (17/72), the posterior MFL 70.8% (51/72) and the two ligaments coexisted 16.7% (12/72). The prevalence of pseudo-tears of the posterior horn of the lateral meniscus was 25 cases. All observed pseudo-tears had either in posteroinferiorly oblique direction (19/25) or in vertical direction (6/25).

CONCLUSIONBased on the location and direction of pseudo-tears and observation in the continuous sagittal plane and the coronal plane, pseudo-tears is easily differentiated from the true tear of the lateral meniscus

Adult ; Female ; Humans ; Knee Injuries ; diagnostic imaging ; surgery ; Knee Joint ; diagnostic imaging ; surgery ; Magnetic Resonance Imaging ; Male ; Menisci, Tibial ; diagnostic imaging ; surgery ; Middle Aged ; Radiography ; Tibial Meniscus Injuries

OBJECTIVETo explore the diagnostic value of 3D-indirectmagnetic resonance (MR) arthrography in detecting meniscustears of the knees.

METHODSForm January 2013 to January 2014, routine plain MR of the knees followed by the 3D-indirect MR arthrography was performed in 42 patients with suspected meniscal tears clinically. There were 31 males and 11 females, with an average age of 38.4 years old (ranged, 21 to 67 years old). The duration of the course ranged from 2 h to 15 d. The clinical symptom was knee pain. All the patients got subsequently arthroscopic examination or operation. The sensitivity and specificity of routine plain MR and 3D-indirect MR arthrography were compared based on the results of arthroscopic examination or operation.

RESULTSThe signal intensity in the area of meniscal tears on image of 3D-indirect MR arthrography was obviously higher than that of routine plain MR. The sensitivity of 3D-indirect MR arthrography was 85.79% (87/102), while the routine plain MR was 52.94% (54/102), and the specificity improved from 67.78%(61/90) to 86.67%(78/90). The difference was statistically significant (χ2 = 25.90, P < 0.01; χ2 = 9.13, P < 0.01).

CONCLUSIONIn comparison with the routine plain MR findings, 3D-indirect MR arthrography demonstrates meniscus tears of the knee with the better sensitivity and specificity.

Adult ; Aged ; Arthrography ; methods ; Female ; Humans ; Imaging, Three-Dimensional ; methods ; Knee Injuries ; diagnosis ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Tibial Meniscus Injuries

Objective To study effect of excessive iodine intake on sodium-iodide symporter(NIS)mRNA and protein expression of breast in lactating rats.Methods60 Wistar rats,having been weaned for one month,were randomly divided into three groups according to their body weights,I.e,①normal iodine(NI,30 rats);②ten fold high iodine(10 HI,15 rats);③one hundred fold high iodine(100 HI,15 rats).Eating food containing iodine of 300μg/L and drinking water of iodine at 5,1845,20 295μg/L,respectively.After fed for 3 months,the rats mated and had offspring,and urine and milk iodine of lactating rats were determined by As-Ce-catalytic spectrophotometric method.Their marmnary glands were sampled at lactation day 10.Then NIS mRNA expression by RT-PCR was determined and NIS protein by immunohistochemistry(SABC)was observed.Results The urine iodine of 10 HI group(3597.5μg/L)and 100HI group(25 404.3μg/L)increased obviously compared with that of NI group(344.7μg/L).The milk iodine of 10HI group(27.1×103μg/L)and 100HI group(191.0×1μg/L)was higher than that of NI group(6.0×103μg/L),but the increased fold of milk iodine was not paralleled with that of urine iodine.Difference of NIS mRNA expression was significant(F=24.19,P＜0.01)among the groups,and the NIS mRNA expression in 10HI(1.250±0.034)and 100HI(1.272±0.039)group were less than that in NI (1.532±0.044)group(P＜0.01).The breast NIS mRNA expression in lactating rats(1.532±0.044)was significantly higher than that in unlactating rats(0.879±0.018,P＜0.01).With the increasing iodine uptake,NIS protein expression decreased.Conclusions The NIS mRNA and protein in rat breasts is down-regulated by excessive iodine intake.So increasing extent of milk iodine concentration is inhibited,which is important to prevent off-spring from getting excessive iodine intake from parental generation.

Daniel Bernoulli (1700-1782) was a famous physicist and mathematician in the 18th century. He made a compelling statistical case for smallpox vaccination. He made important contributions to the knowledge of probability theory and calculus, and was the first person to put forward the question of statistical hypothesis test. At the same time, he applied probability theory to practical problems such as the ratio of male to female births. In the development of epidemiology and biostatistics, he seems to be a brick picker, moving indispensable cornerstone.

William Petty ( 1623－1687) was a great 17th－century economist，statistician and one of the founders of the Ｒoyal Society． He put forward the concept of " political arithmetic" and used the method of " number，weight and measurement" to calculate and analyze the population and economic data of a country． His statistical methods laid the foundation for the development of statistics．

OBJECTIVETo investigate the inhibitory effects of antisense oligonucleotides to different sequences on VEGF gene expression by human hepatoma cells.

METHODSSMMC7721 cells were cultured under normoxic or hypoxic conditions for 24 h, followed by being transfected with different antisense oligonucleotides (A06513 to cap structure, A06514 to translation initiation, A06515 to Exon-3 and A06516 to translation terminal). The total RNAs from the cells were extracted and the VEGF expression were examined with RT-PCR. The relative concentrations of VEGF transcripts in SMMC772 cells from different groups were determined using GAPDH (glyceraldehyde-3-phosphate dehydrogenase) cDNA as internal standard.

RESULTSIn response to the hypoxic challenge, SMMC7721 cells upregulated VEGF mRNA; Comparative to the control (no oligonucleotides), A06513, A06514, A06515, and A06516 had obvious sequence-specific inhibitory effect on VEGF gene expression, with the ratio of VEGF over GAPDH of 0.49+/-0.08, 0.71+/-0.12, 0.72+/-0.11 and 0.86+/-0.12, respectively (F=12.21, P< 0.05). A06513 showed the strongest inhibitory effect (P<0.01).

CONCLUSIONThe antisense oligonucleotides complementary to VEGF cap structure, may become a potential alternative for antisense gene therapy of HCC.

Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms ; genetics ; therapy ; Oligonucleotides, Antisense ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; antagonists & inhibitors ; genetics

Objective To disscus the applicability of three surveillance methods for Aedes albopictus in dengue risk indication. Methods Larval and adult mosquito surveillance were conducted in two villages of Guangzhou by using three vector monitoring methods. Meteorological data and dengue cases were obtained in the same period. Results The population dynamics of larvae and adult mosquitoes were affected by temperature and their overall trend was consistent, but there were differences between the abundance of larvae and adult mosquitoes. From December to February the next year, Breteau index(BI) ranged from 5.66 to 24.53 with a risk of level 1 to 3 for dengue fever, while the mosquito and oviposition positive index(MOI) and adult-mosquito density index(ADI) were 0 to 4.00 and 0 to 1 per man-hour, respectively, indicating no risk for dengue fever. In March, compared with BI which indicated a risk of level 3, MOI were 2.13 and 3.77, respectively, representing for no risk. ADI were 4 to 6 per man-hour, indicating a risk of level 1 and level 2, respectively. After April, the dengue risk level reflected by each indicator became to be consistent. Conclusions The risk of dengue fever indicated by BI is always higher than or equal to the risk indicated by the other two. Compared with BI, MOI and ADI are more sensitively in reflecting the activity of Ae. albopictus.

No abstract available.
Aged, 80 and over ; Anti-Bacterial Agents/pharmacology ; Bacterial Proteins/*genetics ; China ; Drug Resistance, Multiple, Bacterial ; Humans ; Male ; Methyltransferases/*genetics ; Microbial Sensitivity Tests ; RNA, Ribosomal, 16S/genetics/metabolism ; Serratia marcescens/drug effects/*enzymology/*genetics/isolation & purification ; Urinary Tract Infections/diagnosis/microbiology

OBJECTIVETo investigate the diversify of the nuclear pathway of c-Jun NH2-terminal kinases (JNK) during transient brain ischemia/reperfusion injury in hippocampal neuron apoptosis in spontaneously hypertensive rats (SHR) and to test whether the neuroprotection of curcumine on transient brain ischemia/reperfusion injury in SHR is related to the nuclear pathway of JNK.

METHODSMale Wistar-Kyoto (WKY) rats and SHR were randomly divided into five groups (n = 6): WKY sham group (W-Sham), WKY ischemia/reperfusion group (W-I/ R), SHR sham group (S-Sham), SHR ischemia/reperfusion group (S-I/R) and SHR curcumine (a chinese traditional medicine)100 mg/kg treatment group (S-Cur), which were sacrificed at 2 h, 6 h, 24 h, 3 d and 7 d after reperfusion. Global brain ischemic model was established by 4-VO method. The TdT-mediated dUTP nick end labeling (TUNEL) method was used to detect the neuron apoptosis in hippocampal CA1 region. The immunohistochemical method was applied to investigate the expressions of c-jun and c-fos in hippocampal CA1 region.

RESULTSThe expressions of apoptosis and c-jun and c-fos in CA1 region in S-Sham group, W-I/R group and S-I/R group were more than those in W-Sham group (P < 0.05), were significantly increased in S-I/R group than those in W-I/R group (P < 0.05), and were significantly decreased in S-Cur group than those in S-I/R group (P < 0.05).

CONCLUSIONNeuronal apoptosis and the expressions of c-jun and c-fos are more in SHR hippocampal. Global brain ischemia/reperfusion injury induces more expressions of apoptosis in hippocampal neuron in SHR, and the more expressions of c-jun and c-fos may participate in that process. The neuroprotection of curcumine in SHR is related to c-jun and c-fos.

Animals ; Apoptosis ; drug effects ; Brain Ischemia ; metabolism ; pathology ; CA1 Region, Hippocampal ; drug effects ; metabolism ; Curcumin ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Male ; Neurons ; drug effects ; metabolism ; Proto-Oncogene Proteins c-fos ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Reperfusion Injury ; metabolism ; pathology