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Objective:To explore the effect of quantifying food thickness in patients with swallowing difficulty after thyroidectomy.Methods:A total of 70 patients with swallowing difficulty after thyroidectomy in Department of Thyroid and Neck Oncology, Tianjin Medical University Cancer Institute & Hospital, during April 2019 to March 2020, randomly divided into intervention group(35cases) and control group(35cases). The patients in the control group were treated with conventional dietary nursing measures after thyroidectomy. The patients in the intervention group were treated with dietary guidance according to the results of quantifying food thickness and conventional dietary nursing. The incidence of aspiration was compared during hospitalization, the swallowing function were evaluated in the two groups pre intervention and on discharge, and relationship was evaluated between Kubota drinking test level and food consistency grade of safe eating in intervention group.Results:The incidence of aspiration in the intervention group (0) was lower than that in the control group 28.57% (10/35) and the difference was statistically significant ( χ2 value was 11.667, P<0.01). Before intervention the difference in the degrees of Kubota drinking test had no statistical significance between two groups before intervention ( Z value was -0.283, P>0.05). After interventions the degrees of Kubota drinking test was graded as gradeⅠ(1 example), gradeⅡ(4 examples), gradeⅢ(20 examples), grade Ⅳ(9 examples), gradeⅤ(1 example) respectively, meanwhile the degrees of control group was graded as gradeⅠ(0 example), gradeⅡ(1 example), gradeⅢ(16 examples), grade Ⅳ(17 examples), gradeⅤ(1 example) respectively. The difference between two groups was statistically significant ( Z value was -2.170, P<0.05). Moreover, the improvement of swallowing function in the intervention group was larger than that of the control group and the difference in the degree of the swallowing function was statistically significant between two groups ( Z value was -2.029, P<0.05). Before intervention the difference of the score of Eating Assessment Tool-10 had no statistical significance between two groups ( t value was 0.168, P>0.05). After intervention the score of Eating Assessment Tool-10 in the intervention group was 8.43±2.21, the score of Eating Assessment Tool-10 in the control group was 10.06±2.45, the difference had statistical significance between two groups( t value was -2.919, P<0.01). Conclusion:Quantification of fluid food thickness can more accurately and objectively judge the grade of food consistency that patients can eat safely, and it can effectively reduce the incidence of aspiration and contribute to improving the swallowing function in patients with swallowing difficulty after thyroidectomy and can also help them to recovery.
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The evolution, structure and antigenic epitopes prediction of Rana dybowskii antimicrobial peptide dybowskin-1ST were carried out using bioinformatics software available online. Its antibacterial mechanism and structural properties were analyzed, and its activity was verified by applying wound healing assay in mice and bacteriostatic assay in vitro. This provides the theoretical basis for the improvement of parental peptide and the development of novel derivative peptides. The software MEGA_X were used to conduct homology alignment and to construct a phylogenetic tree. The online software ProtParam, ProtScale, PeptideCutter, signal, TMHMM Server were respectively used to predict the physicochemical parameters, hydrophilia/hydrophobicity, shear sites, signal peptides, and transmembrane domains of dybowskin-1ST. The online software SOPMA, Jpred4, DNAstar Protean were used to predict the secondary structure of dybowskin-1ST, and SWISS-MODEL, I-TASSER were used to predict the tertiary structure. ABCpred and SYFPEITHI were respectively used to predict its B-and T-cell epitopes. The effect of dybowskin-1ST on the wound healing was observed on experimental mice. Kirby-Bauer method and dilution method were used to determine the bacteriostatic activity of dybowskin-1ST. The dybowskin-1ST consists of 59 amino acid residues, of which leucine accounts for 16.9%, with a molecular formula of C₃₁₈H₅₁₀N₈₀O₉₃S₂. Its theoretical isoelectric point is 5.10 and the charge is -2. The dybowskin-1ST and dybowskin-1CDYa are closely related phylogenetically. The secondary structure of dybowskin-1ST predicted by the three methods were similar, which consisted of α-helix (44.07%), extended strand (16.95%), β-turns (3.39%), and random coil (35.39%). The prediction of tertiary structure showed that dybowskin-1ST was mainly composed of α-helix, and it was regarded as a hydrophilic protein with signal peptide sequence. Subcellular localization analysis showed that the probability of secreting the mitochondrial targeted peptides was 0.944. Dybowskin-1ST is an extracellular protein with no transmembrane structure region, but contains seven phosphorylation sites, three T-cell epitopes and eight B-cell epitopes. The dybowskin-1ST promoted wound healing and effectively inhibited the growth of Escherichia coli and Staphylococcus aureus. However, it had limited antibacterial activity against fungi and drug-resistant bacteria. Although the structure of dybowskin-1ST is rich in α-helix, the verification experiments showed that its antibacterial ability needs to be enhanced. The reason may be that it is a negatively charged and hydrophilic protein, and amino acid modification with the aim of increasing the number of positive charges and changing the hydrophobicity may be used to obtain derived peptides with enhanced activity.
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Animals , Mice , Amino Acid Sequence , Phylogeny , Pore Forming Cytotoxic Proteins , Protein Structure, Secondary , RanidaeABSTRACT
OBJECTIVE： To establish the method for PCR identification of bullwhip， and to identify the authenticity of bullwhip at the molecular level. METHODS： DNA samples of bullwhip and its counterfeits （donkey whip， pig whip， sheep whip） were extracted and their integrity， purity and concentration were detected. Using GenBank related information， using mitochondrial cytochrome b （Cyt b） gene of bullwhip as target gene， Primer-BLAST online software was used to design specific primer. PCR amplification was performed for whips of different species， and electrophoretic analysis was conducted for the product. PCR products of bullwhip samples were cloned and confirmed by DNA sequencing. The specificity and repeatability of the established PCR method were verified. RESULTS： DNA purity of the bullwhip and its counterfeits was high， and there was no protein or RNA pollution. 1.5％ agarose gel electrophoresis showed that there were obvious target gene bands of bullwhip samples at 200-300 bp， while no corresponding bands appeared in other counterfeit products. The results of DNA sequencing showed that the nucleotide sequence of the gene fragment of bullwhip was 100％ similar to that of the bullwhip in GeneBank. Results of methodological validation showed that established method was specific and reproducible. CONCLUSIONS： The established PCR identification method based on Cyt b gene in the study is simple， rapid， accurate， specific and reproducible， and can meet the requirements of analysis and identification of bullwhip and its counterfeits.
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Objective To investigate the effects of magnesium sulfate and magnesium L-sulfonate on the neurobehavioral response and the expression of induced nitric oxide synthase (iNOS) in neurons after acute cerebral ischemia in rats. Methods One hundred and twelve male Sprague-Dawley ( SD) rats were double-blinded randomly divided into sham-operated group, middle cerebral artery occlusion ( MCAO ) group,MgSO4 treatment group,L-MgT treatment group. Each group was further divided into 6 h,12 h and 24 h subgroups according to the different detection time points. Rat MCAO models were produced following the Longa's method. And the Longa score,limb-placing test,rotarod test,and sticky tape test were performed to evaluate the neurological damage,autonomous movement and coordinate perception in the 24 h subgroup. At the end of the experiment,2,3,5-triphenyltetrazolium chloride ( TTC) was used to evaluate the area of cerebral infarction at 24 h reperfusion. Immunohistochemical staining was employed to determine the altera-tions in iNOS expression in neurons 6 h,12 h and 24 h after reperfusion. Results In behavioral evaluation:Longa score:Normal performance was observed in sham-operated group. Compared with the MCAO group,the scores of MgSO4 treatment group(1. 71±0. 18) and L-MgT treatment group(1. 14±0. 14) were decreased (t=0. 548,3. 873,all P<0. 05),and the score of L-MgT treatment group was lower than that of the MgSO4 group(t=2. 828,P<0. 05). Limb symmetry score:There was no statistically significant difference between MgSO4 group and MCAO group,but there was a statistically significant difference between L-MgT group and MCAO group (t=7. 071,P<0. 05). The roding experiment:The time of MgSO4 group and the L-MgT group were significantly different from that of the MCAO group (t=9. 588,20. 776,P<0. 05),and the time of the L-MgT group was significantly higher than that of the MgSO4 group (t=4. 983,P<0. 05). The right limb strip removal experiment: The time of MgSO4 group and L-MgT group were statistically different from that of MCAO group (t=6. 135,5. 825,P<0. 05),and the time of L-MgT group was increased compared with that of MgSO4 group(t=4. 507,P<0. 05). TTC test:No infarction was formed in the sham group. Compared with MCAO group ((36. 82±1. 35)%),the cerebral infarction volume of MgSO4 group ((17. 39±1. 72)%) and L-MgT group ((10. 81 ± 1. 35)%) significantly decreased, with statistically significant differences ( t=8. 874,11. 105,P<0. 05). Compared with MgSO4 group,cerebral infarction volume in L-MgT group was sig-nificantly reduced,with statistical significance (t=2. 593,P<0. 05). HE staining:There was no statistically significant difference in cell morphology between MgSO4 group and MCAO group at each time point,but the cell morphology of L-MgT group was intact compared with that of MCAO group. INOS staining at 24 h:There was no statistically significant difference in the positive cell density between the MgSO4 group and the MCAO group,but the L-MgT group (cortex:(196. 7±8. 1);striatum:(153. 3±3. 8)) positive cell density was lower than that of the MCAO group (cortex:(375. 0±6. 7),striatum:(358. 3±4. 5)),and the difference was sta-tistically significant (t=11. 113,36. 231,P<0. 05). Conclusion L-MgT may have a significantly protective effect on MCAO rats,and its mechanism may be related to the level of iNOS in neurons.
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Objective To investigate the benefit finding for caregivers of patients with cancer and its influencing factors. Methods Totally 330 couples of cancer patients and caregivers were investigated from October 2014 to September 2015 by convenience sampling method. The cancer patients were investigated with the Hospital Anxiety and Depression Scale. The caregivers of cancer patients were investigated with Benefit Finding Scale, Zarit Caregiver Burden Interview, Medical Outcomes Study Social Support Survey. Results The total score of benefit finding was (59.65 ± 11.12) points. Regression analysis indicated that caregivers' education, religious, informational and emotional support, caregiver burden, patients' depression symptom, cancer stage and self-caring were influencing factors of benefit finding for caregivers (t=-3.689-5.913, P<0.01 or 0.05), which explained 30.0% of the variance. Conclusions The caregivers of patients with cancer have a low level of benefit finding. Oriented by influencing factors of benefit finding, the health workers should enhance targeted interventions, promote positive perception in the process of caring, develop the quality of caring and life.
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Objective To prepare a primary dysmenorrhea self-management behavior scale used for female college students in China.Methods Based on Green health education model,a pool of 40 items was made through literature review as well as semistructured interviews on primary dysmenorrhea in female college sudent.In the following,a primary dysmenorrhea scale for female university students with 20 items was made by considering the advice from five obstetrists and gynecologists and employing a pre-test.Then the primary scale was used for an investigation among 100 female college students with dysmenorrheal and the items were screened by using by item analysis,and a scale with 19 items was decided.The revised scale was finalized by using it among 500 female students with dysmenorrheal in a college in Dalian for a formal investigation,with the reliability and validity tested by factor analysis and correlation analysis.Results The finalized scale was estabhshed containing four dimensions of 19 items.The scale internal consistency Cronbach's α coefficient was 0.91,the index on behaviour,gaining knowledge,treatment compliance and asking for help 0.77,0.88,0.86 and 0.84 respectively.The retest reliability was 0.92 and the index on each dimension was 0.76,0.84,0.83 and 0.77,respectively.The total content validity index of scale content validity index (S-CVI) was 0.93.The content validity index of each dimension,the item cotent validity (I-CVI)was 0.93,0.92,0.92 and 0.94,respectively.By exploratory factor analysis and screen plot,four factors were extracted,with a cumulative contribution rate of 62.90％.The correlation coefficients between the various dimensions were 0.12 to 0.35 and the correlation coefficients between the dimensions and total score were 0.49 to 0.78 (all P ＜0.05).Conclusions Primary dysmenorrhea self-management behavior scales for female college students has a good reliability and validity.It can be used as a tool to evaluate the self management behaviors of female college students in our country.
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Objective To examine the impact of oncology nurses' compassion satisfaction and compassion fatigue on doctor-patient relationship satisfaction.Methods Questionnaire survey was conducted by 388 oncology nurses,the questionnaires include Professional Quality of Life Scale and DoctorPatient Relationship Satisfaction Scale.Results The total scores of compassion satisfaction was 34.22±7.19,the total scores of secondary traumatic was 26.32±6.12,the total scores of job burnout was 25.13±6.08,the total scores of doctor-patient relationship satisfaction was 4.24±0.75.Pearson correlation analysis showed that doctor-patient relationship satisfaction score was positively correlated with oncology nurses' compassion satisfaction (r=0.468,P ＜ 0.05),negatioely correlated with nurses' compassion fatigue factor scores (r=-0.182,-0.531).Regression analysis showed that compassion satisfaction,secondary traumatic,job burnout and night shift frequency were influencing factors of doctor-patient relationship satisfaction,which could explain29.2％ of the variance.Conclusions Improvement of the oncology nurses' compassion satisfaction and compassion fatigue can improve the doctor-patient relationship satisfaction and the quality of nursing service,and create a harmonious medical environment.
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Objective To construct a stable infectious clone of S191 virus strain used for the pro-duction of live attenuated measles vaccine. Methods Full length cDNA of S191 strain and gene fragments encoding nucleocapsid(N),phosphoprotein(P)and RNA polymerase(L)were synthesis and respectively cloned into the vector pVAX1. The 293T cells were respectively transfected with the recombinant expression plasmids and co-cultured with Vero cells. The supernatants of cell culture were collected for identifying res-cued viruses. The indirect immunofluorescence assay was performed for virus identification. The rescued viruses at different passages in Vero cells and the sequences of the rescued viruses were analyzed. Results Restriction enzyme digestion and sequence analysis showed that the recombinant expression plasmids contai-ning the full length cDNA with an artificially engineered mutation at nucleotide 2101(C-A)and gene frag-ments encoding N,P and L proteins of S191 strain were constructed successfully. The N and P proteins were detected in Vero cells with immunofluorescence assay. A cytopathogenic effect on Vero cells was induced by rescued viruses. Conclusion The stable infectious clones of S191 virus used for the production of live at-tenuated measles vaccine were rescued successfully. An approach by using reverse genetics technique for S191 strain study was established which could be used for the development of new chimeric vaccines against measles virus.
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Objective To investigate the distribution of eosinphililic neurons ( ENs),reactive astrocytes ( RAs),and infarction after transient cerebal ischemia,and the time profile of pathomorphological changes.Methods Unilateral forebrain ischemia was induced in Mongolian gerbils by two 10 minutes unilateral common carotid artery occlusions with a 5 hours interval.Laser Doppler flowmetry was used to detect intra-ischemic anterior cortex blood flow.Animals were sacrificed at 24 hours,4 days,2 weeks,4 weeks,16 weeks and the brain were prepared for pathomorphological assay.Results Intra-ischemic laser Doppler flowmetry show significant ischemia during carotid artery occlusion:22.1％ ± 9.5％,26.3％ ± 4.9％,37.5％ ± 3.5％,F =67.219,P ＜ 0.01 ; the decrease was significantly greater in the anterior cortex.ENs appeared in middle and deep layers at 24 hours postischemia,and ENs area extend to all layers of cortex by 4 days.Large areas of high EN density ( ≥80/mm2) evolved to infarcts between 4 days and 4 weeks.Posterior cortex evolved to low EN area ( ＜ 80/mm2) without transformation into infarcts.RAs were consistently distributed in areas with ENs,and RA areas with high EN density were largely transformed into infarcts between 4 days and 4 weeks. Delayed astrocytic death took place in the RA areas with high EN density.Conclusion Density of ENs is an important indicator of delayed astrocytic death and infarction in postischemic tissue.
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Objective To determine LDLR gene mutation in 2 clinically diagnosed FH patients from Hubei province and provide basis for gene diagnosis of FH.Methods Clinical data of 2 FH patients and their parents were collected.The promoter region and exon 1 to exon 18 region of LDLR gene were amplified through PCR and the amplified products were analyzed by forward and reverse DNA sequencing.The mutations were identified after comparison with LDLR gene sequence in GenBank.The pathogenic gene mutations were confirmed according to both genotype and phenotype of FH probands.Results The levels of plasma TC of two probands were 12.79 and 11.98 mmol/L.respectively.No gene mutations were detected in region 3 500 to 3 531 of ApoB100. The mutations of LDLR gene were compound heterozygous mutations. The novel mutation 665G > T detected in the exon 4 of No. 1 proband's LDLR gene was heterozygous missense mutation. The novel mutation 1 358 +32C > T was detected in the exon 9 of No. 1 proband's LDLR gene.The mutations 665G > T ( paternal origin) and 1 358 + 32C > T ( maternal origin) were inherited from the parents. A novel mutation 1 257 C > A was detected in the exon 9 of No. 2 proband's LDLR gene, resulting the presence of a premature termination codon, which was different from 1 257 C > G reported in Belgium.Another heterozygous missense mutation 1 879 G > A was detected in exon 13. They were derived from paternal origin and maternal origin, respectively. Conclusions There are three novel gene mutations:665G >T, 1 358 +32C > T, 1 257C > A found in two probands with compound heterozygous mutations in LDLR respectively. They maybe play a potential role in FH pathogensis.
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Objective To identify the species of brucella rapidly with small fragments of PCR products. Methods The primers were designed according to Brucella spp. repeated insertion sequence IS711 in addition to the specific gene sequences of Brucella melitensis, Brucella abortus and Brucella suis. The small fragments of 3 standard strains and 13 clinical isolates of Brucella were amplified by PCR. Then PCR products were T-A cloned and sequenced. The DNA of standard strains were diluted for sensitivity and stability testing. Results Four specific PCR products were obtained by PCR (63, 67, 81 and 83 bp). The results of T-A cloning and sequencing were in accord with the target gene fragment test. The detection limit of DNA was 1 μg/L. The expected results were achieved with different primers. Conclusion Species identification of Brucella with small fragments of PCR production is a method to identify Brucella strains with great specificity,sensitivity and stability.
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Objective To study the expression and relationship among CD44V6,p16 and PCNA in gastric mucosal lesions with HP infection. Methods Expression of CD44V6,p16 and PCNA were investigated in 114 gastric mucosal lesions by use of immunohistochemistry.HP was examined by both Warthin -Starry method and RUT. Ruselts Comparing HP positive group (P