Objective To investigate the effects of 5'-N-ethylocarbo-xamidoad-enosine(NECA)on cerebral ischemia-reperfusion oxidative injury and the levels of miR-29b and miR-125b in rats.Methods SD rats were divided into sham-operation,model,experimental and positive control groups with 12 rats in each group.Cerebral ischemia-reperfusion model was established in all the other 3 groups except control group.7 days before modeling,sham-operation group and model group were intraperitoneally injected with the same amount of normal saline,and experimental group was injected with 1.5 mg·kg-1 NECA,and positive control group was injected with 2.0 mg·kg-1 nimodipine.The rats in 4 groups were given the drug once a day for 7 days.Cerebral ischemia-reperfusion model was established by middle cerebral artery embolization in all the other 3 groups except sham-operation group.After the modeling was successful,the serum oxidative stress indexes glutathione(GSH),superoxide dismutase(SOD),catalase(CAT),malondialdehyde(MDA)of rats were determined by enzyme-linked immunosorbent assay,and the levels of miR-29b and miR-125b in brain tissue were detected by real-time fluorescence quantitative polymerase chain reaction.Results The SOD levels of sham-operation,model,experimental and positive control groups were(57.82±6.87),(10.75±0.24),(32.24±3.42)and(29.45±4.11)U·mL-1;the levels of GSH were(20.62±3.12),(9.45±1.92),(16.78±1.85)and(15.39±2.02)U·mg-1;the levels of CAT were(3.25±0.42),(0.85±0.11),(2.18±0.34)and(1.98±0.32)U·L-1;the levels of MDA were(0.42±0.09),(1.35±0.42),(0.75±0.06)and(0.72±0.08)nmol·mg-1;the expression levels of miR-29b in brain tissue were 1.02±0.11,0.42±0.02,0.84±0.08 and 0.87±0.10;the expression levels of miR-125b in brain tissue were 1.00±0.11,0.48±0.05,0.75±0.08 and 0.74±0.07,respectively.There were statistically significant differences between sham-operation group and model group(all P＜0.05).Compared with experimental group and positive control group,there were statistically significant differences in the above indexes(all P＜0.05).Conclusion NECA can improve oxidative damage and increase the expression levels of miR-29b and miR-125b in cerebral ischemia-reperfusion rats.

Slow freezing is the most commonly used technique for the cryopreservation of spermatozoa in clinical practice. However, it has been shown to have a negative impact on sperm function and structure. Vitrification as a successful alternative method has been proved to have better protective effects on human embryos, but vitrification of spermatozoa is still subject to low recovery rates. In this study, a modified vitrification method for native spermatozoa was developed. A total of 28 semen samples were included; each sample was divided into three equal parts and assigned to fresh, slow freezing, and vitrification groups. Sperm vitality, motility, morphology, DNA integrity, and acrosome reaction were assessed for each of the groups. The results showed that vitrification achieves better results for several sperm protection parameters than slow freezing; vitrification achieves a higher recovery rate (P < 0.05), motility (P <0.05), morphology (P <0.05), and curve line velocity (P <0.05) than slow freezing. Furthermore, DNA fragmentation was decreased (P <0.05) and better acrosome protection (P <0.05) was exhibited in the spermatozoa after vitrification. Principal component analysis of all sperm parameters revealed that the vitrification cluster was closer to the fresh cluster, indicating that spermatozoa are better preserved through vitrification. In conclusion, while both slow freezing and vitrification have negative effects on sperm function and structure, the vitrification protocol described here had a relatively better recovery rate (65.8%) and showed improved preservation of several sperm quality parameters compared with slow freezing.

Objective:To construct a teamwork model, Partnership Rehabilitation Therapy (PRT), for therapists in critical wards for patients with Corona Virus Disease 2019 (COVID-19), and observe its effect. Methods:PRT had been developed, in which one therapist (main) implementing therapy and another (assistant) monitoring and supporting in the treatment. Eleven COVID-19 patients from infectious critical ward were treated with PRT. The behavior safety of therapists was recorded during the treatment, and the patients were assessed with Borg Index, Cough Score, Miller Sputum Grading and World Health Organization Disability Assessment Schedule (WHODAS) 2.0 before and after treatment. Results:No physiotherapist was infected by COVID-19. Seven times of infection risks were recorded and avoided, and six times of treatment risks were corrected instantly. All the patients improved in Borg Index (P < 0.01), Cough Score (P < 0.05), Miller Sputum Grading (P = 0.02) and WHODAS 2.0 (P < 0.01) after a 1-week physical therapy. Conclusion:Based on the Family International Classifications, a teamwork model is established, which provides a safe and practicable way for rehabilitation for COVID-19 patients in critical wards.

Tung tree (Vernicia fordii) is an economically important woody oil plant that produces tung oil rich in eleostearic acid. Here, we report a high-quality chromosome-scale genome sequence of tung tree. The genome sequence was assembled by combining Illumina short reads, Pacific Bio-sciences single-molecule real-time long reads, and Hi-C sequencing data. The size of tung tree gen-ome is 1.12 Gb, with 28,422 predicted genes and over 73％ repeat sequences. The V. fordii underwent an ancient genome triplication event shared by core eudicots but no further whole-genome duplication in the subsequent ca. 34.55 million years of evolutionary history of the tung tree lineage. Insertion time analysis revealed that repeat-driven genome expansion might have arisen as a result of long-standing long terminal repeat retrotransposon bursts and lack of efficient DNA deletion mechanisms. The genome harbors 88 resistance genes encoding nucleotide-binding sites;17 of these genes may be involved in early-infection stage of Fusarium wilt resistance. Further, 651 oil-related genes were identified, 88 of which are predicted to be directly involved in tung oil biosynthesis. Relatively few phosphoenolpyruvate carboxykinase genes, and synergistic effectsbetween transcription factors and oil biosynthesis-related genes might contribute to the high oil content of tung seed. The tung tree genome constitutes a valuable resource for understanding genome evolution, as well as for molecular breeding and genetic improvements for oil production.

Varus deformation in knee joint is one of the common symptoms caused by unicompartment knee osteoarthritis. Currently, several operations can be used for correcting such deformation, including high tibial osteotomy, unicondylar knee arthroplasty (UKA) and fibulectomy. UKA has been developed for over 60 years, with the advantage of normal knee kinematics restored, less incision, more bony tissue preserved and larger range of motion than total knee arthroplasty (TKA). Therefore, UKA has become a reliable method for treating unicompartment knee osteoarthritis. Fibulectomy is a new kind of surgical technique for treating varus deformation in knee joint, with the advantage of simple operation, low cost and fast recovery. At present, fibulectomy has been widely applied, but its treatment mechanism is still not clear. In this review, two clinical operations UKA and fibulectomy were summarized, and the possible mechanism of fibulectomy for treating unicompartment knee osteoarthritis was proposed from the viewpoint of biomechanics. The author hypothesized that reduction in lateral muscle force after fibulectomy would cause rebalance of the resultant joint moment, therefore, the change of joint contact position and the decrease in joint contact force might be the cause of fibulectomy to release the pain for knee osteoarthritis patients.

BACKGROUNDAnkylosing spondylitis (AS) is a common inflammatory rheumatic disease which lacks satisfactory treatment so far. Sinomenine (SIN) is an alkaloid and has recently been utilized in treating multiple rheumatic diseases including AS in China, but its exact mechanism remains to be explored. This study investigated the alteration of proteome in peripheral blood mononuclear cells (PBMCs) from AS patients.

METHODSThirty AS patients were enrolled in this study. PBMCs from each AS patient were cultured in medium with or without SIN respectively. Then PBMCs proteins from both groups were separated by two-dimensional electrophoresis (2-DE) and analyzed by mass spectrometry (MS). Two differentially expressed proteins were then chosen to be verified using Western blotting.

RESULTSSeven proteins, including a-synuclein (SNCA), calmodulin (CALM), acidic leucine-rich nuclear phosphoprotein 32 family member A (ANP32A), chloride intracellular channel protein 1 (CLIC1), guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1 (GNB1), gelsolin (GSN) and histone H2B type 1-M (HISTH2BM) were over-expressed, while coronin- 1A (CORO1A) was under-expressed in the SIN-treated PBMCs. Further bioinformatics search indicated that the changes of SNCA, ANP32A and CLIC1 pertained to apoptosis, while changes of GSN and CORO1A were associated with both apoptosis and inhibition of immunological function. Subsequently GSN and CORO1A were selected to validate by Western blotting and the results were consistent with those of 2-DE.

CONCLUSIONThere were 8 differentially expressed proteins in the SIN-treated PBMCs, which might shed some light on the mechanism of SIN in the treatment of AS.

Adolescent ; Adult ; Blood Proteins ; analysis ; Blotting, Western ; Cells, Cultured ; Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Leukocytes, Mononuclear ; chemistry ; Male ; Middle Aged ; Morphinans ; pharmacology ; Proteomics ; methods ; Reproducibility of Results ; Spondylitis, Ankylosing ; blood

OBJECTIVETo compare the accuracy of 64-slice computed tomography and coronary angiography in the diagnosis of coronary atherosclerosis.

METHODSThe imaging data of both 64-slice computed tomography and coronary angiography in 65 patients were analyzed retrospectively.

RESULTSAmong the 455 coronary arteries evaluated, the sensitivity, specificity and accuracy of 64-slice computed tomography were 54.6%, 95.1%, and 85.5%, respectively.

CONCLUSION64-slice computed tomography has a good potential in the diagnosis of coronary atherosclerosis with excellent sensitivity and specificity.

Adult ; Aged ; Coronary Angiography ; Coronary Artery Disease ; diagnosis ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Sensitivity and Specificity ; Tomography, Spiral Computed ; methods

OBJECTIVETo develop a 96-microwell plate DNA diagnostic chip for simultaneous detection of 9 major foodborne bacteria.

METHODSType-specific PCR primers labeled with biotin and oligonucleotide probes were designed according to the conservative genes of 9 major foodborne bacteria Staphylococcus aureus, Salmonella spp., Escherichia coli O157:H7 (Stx1 and Stx2), Shigella spp., Listeria monocytogenes, Bacillus cereus, Yersinia enterocolitica, Vibrio cholerae and Vibrio parahaemolyticus. A one-tube multiplex PCR system for simultaneous amplification of these bacteria was established, and the DNA probes were spotted and immobilized in the wells of the plate in 5x5 array format. Stable hybridization system between PCR products and oligonucleotide probes in the microwell was established after condition optimization. Alkaline phosphatase-conjugated streptavidin and NBT/BCIP were used to detect the hybridized PCR products.

RESULTSTwenty standard bacteria strains were used to validate the 96 microwell plate DNA diagnostic chip and highly specific and stable experiment results were obtained. Using this chip assay, the causal pathogen Staphylococcus aureus was identified within 12 h after the sampling from an incident of food poisoning, and the result was consistent with that obtained using conventional bacterial culture and biochemical identification.

CONCLUSIONThe novel 96 microwell plate DNA diagnostic chip allows rapid, accurate, automated and high-throughput bacterial detection and is especially valuable for quick response to such public health emergencies as food poisoning.

Bacteria ; classification ; genetics ; isolation & purification ; DNA, Bacterial ; analysis ; Food Contamination ; analysis ; Food Microbiology ; methods ; Foodborne Diseases ; microbiology ; Humans ; Oligonucleotide Array Sequence Analysis ; methods

OBJECTIVETo developed a multiplex RT-PCR assay for simultaneous screening of type A, B and novel A (H1N1) influenza viruses.

METHODSTwo pairs of universal primers in were designed for amplifying the M gene and NS gene of type A and B influenza viruses, respectively. A pair of specific primers of HA gene was designed to detect novel A (H1N1) influenza virus. A one-step method was used to establish the multiplex RT-PCR system. A blinded experiment was carried out to validate the accuracy of this assay in comparison with the results of real-time fluorescence RT-PCR. The clinical practicability and efficacy of this assay was also evaluated.

RESULTSThe RT-PCR products were analyzed using agarose gel electrophoresis, which yielded distinct bands of the target fragments without non-specific reactions, suggesting the high efficiency and specificity of the multiplex RT-PCR. Blinded study of 50 samples demonstrated a concordance rate of 100%.

CONCLUSIONThis multiplex RT-PCR assay allows one-step simultaneous detection of type A, B and novel A (H1N1) influenza viruses rapidly and accurately, and provides a valuable low-cost screening technique for influenza epidemic monitoring and early diagnosis.

Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; Influenza B virus ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Time Factors ; Viral Matrix Proteins ; genetics ; Viral Nonstructural Proteins ; genetics