Animals ; Antibiotics, Antineoplastic ; administration & dosage ; toxicity ; Apoptosis ; drug effects ; Daunorubicin ; administration & dosage ; toxicity ; Disease Models, Animal ; Female ; In Situ Nick-End Labeling ; Male ; Myocardium ; cytology ; metabolism ; pathology ; ultrastructure ; Neuregulins ; metabolism ; pharmacology ; RNA, Messenger ; metabolism ; Rats ; Rats, Wistar ; Receptor, ErbB-2 ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Objeetive To analyze the effect of treatment of rhegmatogenous retinal detachment(RRD) by scleral bucking as well as the relative risk factors affecting the anatomical reattachment and visual recovery.Methods One hundred and fortyeight patients (148 eyes) with RRD treated by sclera buckling surgery in our hospital during January 2012 to January 2016 were retrospectively analyzed.The rate of postoperative retinal anatomic reattachment,the best corrected visual acuity (BCVA) and complications were observed.Logistic regression analysis was performed to analyze the correlative factors affecting the anatomical reattachment and postoperative vision restoration.R~ults Retinal reattachment achieved in 91.9％ after initial surgery and the final success rate for anatomic reattachment was 97.3％ assessed with ophthalmoscope and fundus photography.But these two rates were assessed with the optical coherence tomography (OCT) were 60.1％ and 80.4％ respectively.Single factor Logistic regression analysis showed that retinal detachment was affected by multiple breaks and Grade C1 PVR(all P ＜0.05);Single factor Logistic regression analysis showed that preoperative BCVA,course of disease,retinal detachment range,macular involvement or not had an impact on the postoperative recovery of BCVA (all P ＜ 0.05),preoperative age,refractive status,releasing retinal fluid or not,intravitreal gas injection,combined scleral buckling,and postoperative subretinal fluid,all of these factors had no effect on BCVA recovery after surgery (all P ＞ 0.05).And through multiple factors Logistic regression analysis,preoperative BCVA was an independent risk factor for BCVA recovery after surgery (P ＜ 0.05).Conclusion Scleral bucking is an effective technique for managing RRD,but multiple breaks and Grade C1 PVR are significant risk factors for anatomic.Preoperative BCVA,course of disease,retinal detachment range,macular involvement or not have the impact on the BCVA recovery after scleral buckling,and the preoperative BCVA is the key factor.Early diagnosis and early treatment as well as protecting the preoperative visual acuity can improve prognosis.

Objective To investigate the biological behavior and expression of intergrin-linked kinase (ILK) after exposure to transforming growth factor beta 2 (TGF-β32) in human Tenon fibroblasts (HTFs).Methods The primary ceils of HTFs were cultured by tissue attached culture method and identified by immunofluorescence analysis with Vimentin and keratin.The proliferation levels of HTFs induced by different concentrations of TGF-β2 were analyzed by MTT.The α-smooth muscle actin (α-SMA) and ILK mRNA expression were analyzed by quantitative Real-time PCR.The protein expression of α-SMA,ILK and E-cadherin were analyzed by Western Blot,and the protein expression of α-SMA and E-cadherin were also analyzed by immunofluorescence staining.Results MTT analysis showed that the optical density levels of 5.0 μg · L-1 and 10.0 μg · L-1 TGF-β2 were significantly higher than those of 0.1 μg · L 1,1.0 μg · L-1 and the control after exposure for 48 hours and 72 hours,and all these optical density levels were significantly higher than that for 24 hours (all P ＜0.05).The expression of α-SMA and ILK mRNA increased significantly when cells were treated with 5.0 μg · L-1 TGF-β2 for 48 hours in comparison with the control group (all P ＜ 0.05).The protein of α-SMA,ILK and E-cadherin were expressed both in TGF-β2 treated groups and control group,and TGF-β2 up-regulated the expression of them.There were significant differences when compared with the control group (all P ＜ 0.05).Immunofluorescent staining showed that α-SMA and E-cadherin were detected in TGF-β2 treated groups.α-SMA expressed in cytoplasm,while E-cadherin both in cytoplasm and nucleus.Conclusion TGF-β2 can induce the proliferation,transdifferentiation and adhesion of HTFs,and up-regulate the expression of ILK in vitro,suggests that ILK may play a role in the process of scar formation after glaucoma filtration surgery.

OBJECTIVETo study the effect of H2O2 on the morphological pattern,vitality,proliferation,cycle period of rabbit intervertebral disc nucleus pulposus cells.

METHODSTen New Zealand white rabbits (2 to 3 kg, female) were used for isolating nucleus pulposus cells under sterilized condition. The culture solution with 15% FBS and DMEM/F12 (1:1) was applied for cell cultivation. After 90% cell fusion, the first generation was obtain and stimulated by H2O2 with different concentrations of 0 micromol/L (control group), 130 micromol/L,216 p.mol/L,360 Ipmol/L, 600 micromol/L,and 1000 micromol/L.

RESULTSCompared with the control group, there was little difference of the biological property (P>0.05) in 130 micromol/L and 216 micromol/L H202-treated groups. When the concentration of H2O2 attained 360 micromol/L, 600 micromol/L, and 1 000 micromol/L, the cells suffered aging,with increased cell vacuoles,decreased proliferation,and aging:related increase of 13-galactosidase dyeing. The cell cycle of many nucleus pulposus cells was blocked in G1 stage other than entering S stage. With increasing H2O2 concentrations, the aging degree was increased.

CONCLUSIONA certain concentration of H202 could induce early aging of nucleus pulposus cells,resulting in biological abnormalities of these cells.

Animals ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cellular Senescence ; drug effects ; Female ; Hydrogen Peroxide ; pharmacology ; Intervertebral Disc ; cytology ; drug effects ; Oxidative Stress ; Rabbits

OBJECTIVETo determine the optimal drill area on the footplate with the 3D measurements of the stapes and the vestibular end organs.

METHODSFour temporal bones were extracted from the fresh cadavers and undecalcified polymer-embedded. After serially sectioning, image processing and the 3D precisely reconstruction, a local Cartesian coordinates was established in which the tympanic surface of the footplate was supposed to be XY plane and the Z coordinate axis passed through the central point of the footplate and was vertical to the XY plane. The configurations of the utricle and saccule were delineated quantitatively, and then any distance between one point on the surface of the footplate and another point on the surface of the utricle or saccule and its orientation can be measured.

RESULTSThere was a "V" shaped cleft between the utricle and the saccule. The angle of the" V" shaped cleft was 50.31 +/- 19.90 (17.00 - 68.00) degrees. The apex of the cleft directed anterosuperiorly and approached the footplate center, while beneath the posteroinferior part of the footplate was an open and deep area. The vertical distance from the center point of the footplate to the vestibular end organs was (2.20 +/- 0.548) mm, the maximum of 3.0 mm and the minimum of 1.6 mm.

CONCLUSIONSThe posterior and inferior quadrant of the footplate may be the optimal drill area for the fenestra.

Adult ; Humans ; Imaging, Three-Dimensional ; Saccule and Utricle ; anatomy & histology ; Stapes ; anatomy & histology ; Temporal Bone ; anatomy & histology

A binary plant expression vector, pCM12-slm, carrying the aroAM12 mutant gene encoding bacterial 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) and the Bts1m recombinant gene consisting of 331 N-terminal amino acids of CryIAc and 284 C-terminal amino acids of CryIAb has been constructed. The truncated Bts1 gene was fused with the PR1b signal peptide sequence and expressed in tobacco plants under the control of 2E-CaMV35S promoter and the omega (omega) translation enhancer sequence from tobacco mosaic virus. The mutant aroAM12 was fused with the transit sequence of tobacco EPSPS and expressed in tobacco plants under the control of the CaMV35S promoter. Tobacco leaves were transformed with Agrobacterium tumefaciens LBA4404 harboring the pCM12-slm plasmid, and the transgenic plants were selected directly on medium containing the herbicide. Forty glyphosate resistant plants were regenerated, with a transformation frequency of 27%. Transgenic plants were initially assessed for glyphosate resistance by placing leaf discs on shoot induction media containing the herbicide. Rooted plantlets, propagated from selected transgenic tobacco, were transferred to soil in a greenhouse and tested for glyphosate resistance by spraying them with Roundup at a commercial recommended dose. The glyphosate resistance assay indicated that all the transgenic plants showed highly resistant to the herbicide. The PCR assay showed that the aroAM12 gene was present in all of the 40 T0 transfer plants, and Bts1m genes present in 28 of 40 of the transgenic plants. Southern blot analysis further confirmed that the copy number of the transgenes varied from one to three copies in different transgenic plants. Northern blot and immunodot blot showed that the aroAM12 and Bts1m genes were expressed at the transcription and translation levels. Transgenic plants containing both the aroA M12 and Bts1m genes were further assessed for insect resistance. Tobacco leaves of T0 transgenic plants were infested with tobacco bollworm H. assulta larvae for 6 days. The result (table 1) showed that the survival rate of insect larvae was between 0-10%, and the growth of insect larvae was seriously inhibited, suggesting pCM12-slm as a dual functional vector with potential application in breeding of glyphosate and insect resistance transgenic plants.
Agrobacterium tumefaciens ; genetics ; Animals ; Blotting, Northern ; Blotting, Southern ; Genetic Vectors ; genetics ; physiology ; Herbicides ; pharmacology ; Moths ; pathogenicity ; Plant Diseases ; parasitology ; Plant Leaves ; drug effects ; genetics ; parasitology ; Plants, Genetically Modified ; drug effects ; genetics ; parasitology ; Polymerase Chain Reaction ; Tobacco ; drug effects ; genetics ; parasitology

The Yellow Fever (YF) vaccine, an attenuated yellow fever 17D (YF-17D) live vaccine, is one of the most effective and safest vaccines in the world and is regarded as one of the best candidates for viral expression vector. We here first reported in China the construction and characterization of the recombinant expression vector of yellow fever 17D which contained the proteinase 2A fragment of foot-and-mouth disease virus (FMDV). Three cDNA fragments representing the full-length YF-17D genome, named 5'-end cDNA (A), 3'-end cDNA (B) and middle cDNA (C), were obtained by reverse transcription polymerase chain reaction (RT-PCR), together with the introduction of SP6 enhancer, necessary restriction sites and overlaps for homologous recombination in yeast. Fragment A and B were then introduced into pRS424 in turn by DNA recombination, followed by transfection of fragment C and the recombinant pRS424 containing A and B (pRS-A-B) into yeast. A recombinant vector containing full length cDNA of YF-17D (pRS-YF) was obtained by screening on medium lack of tryptophan and uracil. A recombinant YF-17D expression vector containing FMDV-2A gene fragment (pRS-YF-2A1) was then constructed by methods of DNA recombination and homologous recombination in yeast described above. In vitro transcription of the recombinant vector pRS-YF-2A1 was then carried out and introduced into BHK-21 cells by electroporation. Results of indirect immunofluorescence assay (IFA) and titer determination showed a stable infectious recombinant virus was gotten, whose features such as growth curve were similar to those of the parental YF-17D. The results suggest that the recombinant vector pRS-YF-2A1, by introduction of heterogenous genes via 2A region, is potential to be an effective live vaccine expression vector.
Animals ; Cell Line ; Cloning, Molecular ; Cricetinae ; Epitopes ; immunology ; Foot-and-Mouth Disease ; prevention & control ; Foot-and-Mouth Disease Virus ; genetics ; immunology ; Genetic Engineering ; Genetic Vectors ; Recombination, Genetic ; Saccharomyces cerevisiae ; genetics ; metabolism ; Vaccines, Attenuated ; Viral Vaccines ; genetics ; immunology ; Yellow fever virus ; genetics ; immunology

To establish a sensitive and specific HPLC method for quality control of Radix Paeoniae Alba, HPLC method was applied for quality assessment of Radix Paeoniae Alba. HPLC analysis was performed on a Symmetry C18 column (250 mm x 4. 6 mm ID, 5 microm, Waters, USA). The mobile phase consisted of acetonitrile (solvent A) and water containing 0.1% (v/v) phosphoric acid (solvent B) at a constant flow rate of 0.8 mL x min(-1). An increasing linear gradient (v/v) of solvent A was used (t/min, % A): (0,10), (5,10), (25,15), (45, 22), (46, 65), (50, 80) and (60, 80). The column temperature was set at 25 degrees C. The chromatograms were monitored at 230 nm and the on-line UV spectra were recorded in the range of 190 - 400 nm. The HPLC chromatographic fingerprinting of Radix Paeoniae Alba, showing 11 characteristic peaks, was established from 28 lots of Radix Paeoniae Alba. The areas of main chromatographic peaks were found to complied with the following rule: paeoniflorin > 1, 2, 3, 4, 6-penta-O-galloyl-glucos > albiflorin > methyl gallate > other compounds. The chromatographic fingerprinting of Radix Paeoniae Alba with high specificity can be used to control its quality and assure lot-to-lot consistency.
Benzoates ; analysis ; Bridged-Ring Compounds ; analysis ; China ; Chromatography, High Pressure Liquid ; methods ; Ecosystem ; Glucosides ; analysis ; Mass Spectrometry ; methods ; Monoterpenes ; Paeonia ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Reproducibility of Results

Abstract: Objective　To analyze the pathogenic characteristics and epidemiological significance of human plague related strains in Qinghai Province in recent 30 years, so as to provide scientific basis for on-the-spot disposal and prevention and control measures of plague outbreak in Qinghai Province. Methods　A total of 35 strains of Yersinia pestis isolated from 29 typical human plague outbreaks in Qinghai Province from 1980 to 2011 were selected and studied by biochemical fermentation experiments. Virulence factors detection of Fraction 1 antigen (Fra1), virulence antigen (VW), pigmentation (Pgm) and Yersinia pestis Ⅰ (PstⅠ), determinants and genotyping of differential regions (DFRs) were used to study the pathogenic characteristics. At the same time, according to the epidemic situation of human and animal plague in Qinghai Province in recent years, the current situation of plague prevention and control and epidemic characteristics were analyzed. Results　The biotypes of 35 strains of Yersinia pestis were classical, and the biotypes of 29 strains (82.86%) were of Qinghai-Tibet Plateau type, mainly distributed in southern Qinghai and around lake areas, 2 strains (5.71%) belonged to Qilian Mountains type, mainly distributed in Qilian mountains, and 6 genotypes were identified by DFR. Among them, 16 were type 5, 12 were type 8, 2 were type 10, 1 was type 36, 3 were type 30 and 1 was type 1b, the strains of type 5 and 1b were mainly distributed around the lake and the southern foot of Qilian Mountains, while the strains of type 8, 10, 36 and 30 were mainly distributed in the southern part of Qinghai. Conclusions　The pathogen of Yersinia pestis in Qinghai Plateau has complex biochemical types, the epidemic situation among animals is continuous year after year, the situation of prevention and control is serious, the occurrence and prevalence of plague seriously endanger people's health and social development, so it is necessary to do a solid job in the prevention and control of plague to ensure the safety of people's lives.

Purpose: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the pathogen of coronavirus disease 2019. Diagnostic methods based on the clustered regularly interspaced short palindromic repeats (CRISPR) have been developed to detect SARSCoV-2 rapidly. Therefore, a systematic review and meta-analysis were performed to assess the diagnostic accuracy of CRISPR for detecting SARS-CoV-2 infection. Materials and Methods: Studies published before August 2021 were retrieved from four databases, using the keywords “SARS-CoV-2” and “CRISPR.” Data were collected from these publications, and the sensitivity, specificity, negative likelihood ratio (NLR), positive likelihood ratio (PLR), and diagnostic odds ratio (DOR) were calculated. The summary receiver operating characteristic curve was plotted for analysis with MetaDiSc 1.4. The Stata 15.0 software was used to draw Deeks’ funnel plots to evaluate publication bias. Results: We performed a pooled analysis of 38 independent studies shown in 30 publications. The reference standard was reverse transcription-quantitative PCR. The results indicated that the sensitivity of CRISPR-based methods for diagnosis was 0.94 (95% CI 0.93–0.95), the specificity was 0.98 (95% CI 0.97–0.99), the PLR was 34.03 (95% CI 20.81–55.66), the NLR was 0.08 (95% CI 0.06– 0.10), and the DOR was 575.74 (95% CI 382.36–866.95). The area under the curve was 0.9894. Conclusion Studies indicate that a diagnostic method based on CRISPR has high sensitivity and specificity. Therefore, this would be a potential diagnostic tool to improve the accuracy of SARS-CoV-2 detection.