Objective To explore prevention and cure effects of the freeze-dried human fibrin glue as the way of the lacrimal duct embolization on xerophthalmia in perimenopausal female rabbit.Methods A total of 72 female rabbits,after anti infection treatment and were cut off third eyelid,were made into perimenopausal xerophthalmia rabbit models.After surgery,all of these rabbits were randomly divided into 6 groups (12 rabbits per group):No treatment group after surgery (group A),PBS prevention group (group B),freeze-dried human fibrin prevention group (group C);no treatment group after modeling (modeling time:Two weeks after surgery,group D),PBS treatment group (group E),freeze-dried human fibrin treatmentgroup (group F).The Schirmer test (SIT),corneal fluorescein (FL) and corneal confocal microscope were performed before and 2 weeks,4 weeks,6 weeks after injection.Results There were statistical differences in FL score and SIT in group A,group B and group C among different time points (F =27.346,10.608;P =0.000,0.001);There were statistical differences between FL scores and SIT among three groups (F =7.579,6.786;P =0.002,0.007);There was significant difference in FL scores and SIT trends among three groups(F =44.897,3.424;P =0.000,0.045).The FL score and SIT of group D,group E and group F were significantly improved after treatment for 2 weeks,4 weeks and 6 weeks,the differences were statistically significant (t =2.906,3.654,4.504;P =0.022,0.017,0.013.t =4.573,5.759,7.231;P =0.032,0.019,0.008);The difference between FL score and SIT in group E and group F was statistically significant after treatment (t =2.776,4.124,5.324;P =0.032,0.026,0.017.t =1.969,3.122,4.324;P =0.038,0.023,0.009).After injection of 6 weeks,the epithelial basal cells (F =17.306,P =0.002) and inflammatory cells (F =34.024,P =0.000) of group A,B and C were significant changed,the differences were statistically significant.After injection of 6 weeks,the epithelial basal cells (F =3.749,P =0.042)and inflarnmatory cells(F=8.806,P =0.005) of group D,E and F were significant changed,the differences were also statistically significant.Conclusion Lacrimal duct embolization with freeze-dried human fibrin glue is effective for the xerophthalmia.

Objective To analyze the thickness of cornea and corneal epithelium in healthy subjects by optical coherence tomography angiography (OCTA).Methods Totally 100 healthy subjects aged between 20 and 30 years were analyzed by OCTA technique.Using AngioVue OCTA system of retinal imaging mode,and using SSADA algorithm for imaging,the cornea and the corneal epithelium in the central corneal diameter range of 9 mm were measured.The differences of corneal and corneal epithelial thickness in different gender regions were compared.Results In the male and female group,the corneal central total thickness were (559.92 ±33.26) μm and(540.06 ±31.63)μm,and the corneal epithelial thickness were(57.78 ±4.88) μm and(56.88 ±4.57) μm,The total central corneal thickness and central corneal epithelial thickness of the male were greater than those of the female,the difference was statistically significant (t =3.06,2.10;all P ＜ 0.05).The cornea of male was the thickest at S5,S7 and SN9,there were significant differences at S5 and S7 compared with female (t =2.93,2.83;all P ＜ 0.05);The female cornea was the thickest at S5,SN7 and SN9,and the difference was significant at S5 compared with male.The cornea of male subjects was the thinnest at IT,which was statistically significant only at IT5 compared with female subjects in the same area (t =2.02,P ＜ 0.05);The cornea of female subjects was the thinnest at T5,IT7 and T9,which was statistically significant only at T5 and T9 compared with male subjects in the same region (t =2.63,2.20;all P ＜ 0.05);There was significant difference in corneal thickness between male and female at ST (t =3.1 1,2.79,2.33;all P ＜ 0.05).The corneal epithelium was the thickest at IT5,I7,and I9,and the lowest at S5,S7 and S9,and there was no significant difference compared with female in the same region (all P ＞ 0.05).The corneal epithelium of female at the IT5,T7,N9 were the thickest,SN5,S7,S9 were the thinnest;Except for M2 and SN5,there was no significant differences in corneal epithelium between male and female groups (all P ＞ 0.05).Corneal central epithelium accounted for the largest percentage of total corneal thickness,and gradually decreased from inside to outside.Conclusion OCTA can be used to measure the thickness of corneal and corneal epithelial regions.

In proteins of thermophilic bacteria, Gly is tend to be replaced by Ala and Lys is tend to be replaced by Arg to adapt the high temperature. In order to improve the thermal stability of phenylalanine hydroxylase (PAH) from Chromobacterium violaceum, all the Gly on PAH were mutated to Ala and Lys to Arg. Positive mutant enzymes with improved thermal stability were selected, followed by combined mutation and characterization. The results revealed that half-lives of K94R and G221A mutants at 50 °C were 26.2 min and 16.8 min, which were increased by 1.9-times and 0.9-times than the parent enzyme (9.0 min). The residual activity of K94R/G221A mutant was improved to 65.6% after keeping at 50 °C for 1 h, which was 6.6 time higher than the parent enzyme (8.6%). Circular dichroism (CD) spectroscopy revealed that Tm values of the parent enzyme, K94R, G221A and K94R/G221A were 51.5 ℃, 53.8 ℃, 53.1 ℃ and 54.8 ℃, respectively. According to the protein structure simulation, the two mutations were located on flexible loop. In the K94R mutant, the mutated Arg94 on the surface of the enzyme formed an extra hydrogen bond with Ile95, which stabilized the located loop. In the G221A mutant, the mutated Ala221 formed hydrophobic interaction with Leu281, which could stabilize both the loop and flexible area of the C-terminus of G221A. The results not only provided a reference for protein modification on thermal stability, but also laid the foundation for application of phenylalanine hydroxylase in the field of functional foods.
Bacterial Proteins ; biosynthesis ; genetics ; Chromobacterium ; enzymology ; Enzyme Stability ; Hot Temperature ; Kinetics ; Mutagenesis, Site-Directed ; Mutation ; Phenylalanine Hydroxylase ; biosynthesis ; genetics ; Protein Engineering

Objective: To investigate the predictive value of neutrophil-to-lymphocyte Ratio (NLR) in 30-day mortality of patients with acute paraquat poisoning. Methods: We respectively reviewed the clinical parameters of 115 patients with acute paraquat poisoning. They were divided into survival (n=64) and non-survival (n=51) groups based on their 30-day outcome. Multivariate logistic regression was performed to identify risk factors of 30-day mortality. Receiver operating curve (ROC) test was applied to analysis to the predictive value of NLR in 30-day mortality ofacute paraquat poisoning patients. The correlations between NLR and severity index of paraquat poisoning (SIPP) were analyzed using Spearman’s rank correlation coefficient. Results: Of the 115patients included in the study, 54 (46.96%) patients were males and 61 (53.04%) were females with a mean age of 38.96±13.58 years. The total mortality in 30-day was 44.35% (51/115) . The NLR at admission was an independently risk factor of 30-day mortality of patients with acute paraquat poisoning (OR 1.477, 95%CI 1.035-2.107, P<0.05) . The NLR to predict the death of the area under the ROC curve was 0.894 (95%CI: 0.8212-0.9663, P<0.01) ; the optimal cutoff threshold was 11.71; the sensitivity was 71.79% and the specificity was 94.29%; the positive predictive value was 93.33%and negative predictive value of 75.00%. Meanwhile, the positive likelihood ratio was 12.57 and the negative likelihood ratio was 0.30. The NLR was significantly associated with SIPP (Spearman rho 0.525; P<0.01) and it was significantly higher in patients with SIPP of ten or higher than in those with an SIPP less than 10 (15.02±12.40 vs. 6.19±2.54, P<0.05) . Conclusion The increased NLR at admission was an independently risk factor of 30-day mortality of patients with acute paraquat poisoning and it was significantly correlated with SIPP score. Therefore, NLR was useful for predicting prognosis of patients with acute paraquat poisoning.

AIM:To analyze the mechanism of tumor-associated macrophage (TAMs) in the development of cervical cancer and to investigate its correlation with Th1/Th2 and Th17/CD4+ CD25+ Foxp3 + Treg.METHODS:Twenty seven cases of cervical cancer and 53 cases of cervical intraepithelial neoplasias (including 22 cases of CIN Ⅱ and 31 cases of CIN Ⅲ) were selected as subjects.The venous blood of patients before treatment was extracted to detect Th1/Th2 and Th17/CD4 + CD25 + Foxp3 + Treg with flow cytometry,and detect serum IFN-γ,IL-4,IL-17A,IL-17F,TGF-β1 and IL-10 levels with ELISA Kits.Furthermore,pathological tissues were extracted during operation,and its TAMsCD68 expression was detected by immunohistochemistry technique.RESULTS:The Th1/Th2 and Th17/CD4 + CD25 + Foxp3 + Treg of cervical cancer were both lower than those of CIN Ⅲ,and those of CIN Ⅲ were both lower than CIN Ⅱ,the difference between groups had statistical significance (P ＜ 0.05).The serum IFN-γ,IL-4,IL-17A,IL-17F,TGF-β1 and IL-10 levels of cervical cancer were all higher than those of CIN Ⅲ,and those of CIN Ⅲ were all higher than CIN Ⅱ,the difference between groups had statistical significance (P ＜ 0.05).The TAMsCD68 expression level of cervical cancer was higher than that of CIN Ⅲ,and that of CIN Ⅲ was lower than CIN Ⅱ,the difference between groups had statistical significance (P ＜ 0.05).The correlation analysis results showed TAMsCD68 expression level had negative correlations with Th1/Th2,Th17/CD4+CD25+ Foxp3+ Treg,and serum IL-17A level,whereas presented positive correlations with serum IL-10 and IL-4 level (P ＜ 0.05).CONCLUSION:TAMs is closely related with Th1/Th2 and Th17/CD4 + CD25 + Foxp3 + Treg in cervical cancer,and possibly is mediating the occurrence and development of cervical cancer through influencing the balance of these two systems.

Based on the resutls of literature review and interviews of experts, two rounds of Delphi surveys were conducted. The mean, importance ratio, coefficient of variation and coordination coefficient were used for assessment of survey from multiple perspectives, and finally form a framework model of factors affecting the efficacy of Tuina therapy. A total of 37 experts were selected for questionnaire surveys, the positive coefficients of experts' participatation in the first round and second round were 92.5% and 80.0%, respectively. The overall coordination coefficient in the second round is 0.68. The items were included into the consensus meeting if the importance ratio of items were equal to and more than 80%. After the expert consensus meeting, 22 items were included to form a framework model of factors affecting the efficacy of Tuina therapy, and summarized as 5 major influencing factors, including diagnostic factors, treatment factors, prognostic factors, patient factors, and doctor-patient communication. This framework can guide and help young Tuina practitioners to improve clinical efficacy. It is also clearly pointed out that the effect of Tuina for pain is not only related to disease diagnosis or manipulation, but also related to home exercise, health care, and doctor-patient communication.

Objective:To survey on the status quo of the role cognition and job burnout among family doctor team members in Shenzhen Futian district.Methods:A survey on the role cognition and job burnout of family doctor members was conducted in October 2022. One family doctor team was randomly selected from each of the 92 community health service centers in Futian district of Shenzhen city, and all members of the selected family doctor teams were included in the survey. The Chinese version of the Maslach Burnout Inventory-Human Services Survey was used in the study, and the influencing factors of role cognition and job burnout, as well as the impact of role cognition on job burnout were analyzed.Results:A total of 361 family doctor team members were included in the survey, The scores of the Role Ambiguity and Role Conflict Scale were (35.25±5.21) and (27.03±13.73), respectively. In the Occupational Burnout Scale, the scores of Emotional Exhaustion, Dehumanization, and Personal Achievement Scale were (17.89±6.82), (6.51±2.54), and (30.95±5.70), respectively. There were significant differences in role ambiguity and job burnout among family doctor team members with different ages, professions, and years of work ( P<0.05). Team members aged≤30, general practitioners, and working time ≤5 years had higher scores for role ambiguity, emotional exhaustion, dehumanization, and lower personal achievement ( P<0.05). The score of role ambiguity was negatively correlated with the degree of emotional exhaustion and dehumanization ( r=-0.544, -0.352), and positively correlated with personal achievement ( r=0.473). The score of role conflict was positively correlated with the degree of emotional exhaustion and dehumanization ( r=0.352, 0.376), and negatively correlated with personal achievement ( r=-0.440). Regression analysis showed that the role cognition was significantly associated with the emotional exhaustion, dehumanization, and personal achievement ( F=10.28-85.12, P<0.01). Conclusion:Active and effective measures should be taken to improve the role cognition and reduce job burnout of family doctor team members in primary health care institutions.

ObjectiveTo evaluate the effects of transcranial direct current stimulation (tDCS) on post-stroke aphasia (PSA). MethodsLiteratures about tDCS for PSA were retrieved from PubMed, Web of Science, Cochrane library, Embase CNKI and Wanfang Data, until December, 2021. The contents of the author, publication time, subjects, intervention methods and time, main outcome indicators and conclusions were extracted. ResultsA total of 1 026 articles were returned and 45 articles were finally included. The publication time was from 2010 to 2021. The subjects of the study were patients with PSA. The main outcome indicators were the various speech scales and their sub-items. For most of the trials, scores of the scales were better in the experimental groups than in the control groups after treatment, and a few found little improvement in Boston Diagnostic Aphasia Examination, or only improved in complex language tasks. tDCS can improve the repetition, naming, spelling and verbal fluency of PSA patients, which might relate with electrode placement, current magnitude, duration, intensity and left cortical integrity. ConclusiontDCS is effective on different language sub-items of PSA, and various with the program of tDCS.

Objective:To establish a clinical diagnostic scoring model for preoperative predicting hepatocellular carcinoma (HCC) microvascular invasion (MVI) based on gadolinium-ethoxybenzyl-diethylenetriamine pentacetic acid (Gd-EOB-DTPA) enhanced MRI, and verify its effectiveness.Methods:From January 2014 to December 2020, a total of 251 cases with pathologically confirmed HCC from Tianjin First Central Hospital and Jilin University First Hospital were retrospectively collected to serve as the training set, while 57 HCC patients from Tianjin Medical University Cancer Hospital were recruited as an independent external validation set. The HCC patients were divided into MVI positive and MVI negative groups according to the pathological results. The tumor maximum diameters and apparent diffusion coefficient (ADC) values were measured. On the Gd-EOB-DTPA MRI images, tumor morphology, peritumoral enhancement, peritumoral low intensity (PTLI), capsule, intratumoral artery, intratumoral fat, intratumoral hemorrhage, and intratumoral necrosis were observed. Univariate analysis was performed using the χ 2 test or the independent sample t-test. The independent risk factors associated with MVI were obtained in the training set using a multivariate logistic analysis. Points were assigned to each factor according to the weight value to establish a preoperative score model for predicting MVI. The receiver operating characteristic (ROC) curve was used to determine the score threshold and to verify the efficacy of this scoring model in predicting MVI in the independent external validation set. Results:The training set obtained 98 patients in the MVI positive group and 153 patients in the MVI negative group, while the external validation set obtained 16 patients in the MVI positive group and 41 patients in the MVI negative group. According to logistic analysis, tumor maximum diameter>3.66 cm (OR 3.654, 95%CI 1.902-7.018), hepatobiliary PTLI (OR 9.235, 95%CI 4.833-16.896) and incomplete capsule (OR 6.266, 95%CI 1.993-9.345) were independent risk factors for MVI in HCC, which were assigned scores of 3, 4 and 2, respectively. The total score ranged from 0 to 9. In the external validation set, ROC curve analysis showed that the area under the curve of the scoring model was 0.918 (95%CI 0.815-0.974, P=0.001). When the score>4 was used as the threshold, the accuracy, sensitivity, and specificity of the model in predicting MVI were 84.2%, 81.3%, and 85.4%, respectively. Conclusions:A scoring model based on Gd-EOB-DTPA-enhanced MRI provided a convenient and reliable way to predict MVI preoperatively.

Objective:To explore the feasibility, accuracy and reproducibility of a novel, fully automated three-dimensional echocardiography right ventricular(RV) quantification software(3D Anto RV) to evaluate the RV volume and RV ejection fraction (RVEF) using artificial intelligence in patients after heart transplantation (HT) comparing with the gold reference-cardiac magnetic resonance (CMR).Methods:Forty-six patients after HT who were scheduled for echocardiogram at their routine follow-up examinations and also agreed to undergo CMR examination within the following 24 hours in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from October 2018 to June 2019 were prospectively included. The right ventricular end-diastolic volume (RVEDV), right ventricular end-systolic volume (RVESV), right ventricular stroke volume (RVSV) and RVEF of HT patients were measured by CMR 3D Auto RV and conventional semi-automated three-dimensional echocardiography RV quantification software (Tomtec 4D RV function 2.0). The results of the 3D Auto RV and conventional semi-automated Tomtec were respectively compared with CMR using paired two-tailed student′s t-tests, Pearson correlation coefficients and Bland-Altman analyses. Results:The feasibility of the 3D Auto RV was 87%.The fully automated analysis realized in 27 (59%) patients by 3D Auto RV and the analysis time required only (12±1)s. The results of the remaining 19 (41%) patients needed manual adjustment and the mean analysis time in manual adjustment was also <2 min that was shorter than the conventional semi-automated three-dimensional echocardiography RV quantification software[(108±15)s vs (160±34)s, P<0.001]. For the results of RV volumes: There were good correlations between the 3D Auto RV and CMR, conventional semi-automated Tomtec and CMR for the measurements of RVEDV, RVESV and RVSV ( r=0.77-0.84, all P<0.001). In addition, compared with CMR, although there were significantly underestimated RV volumes by the 3D Auto RV and conventional semi-automated Tomtec, the negative bias was smaller in the 3D Auto RV than the conventional semi-automated Tomtec. For the results of RVEF: the corresponding RVEF derived from 3D Auto RV and CMR showed an excellent correlation and consistency ( r=0.84, P<0.001; bias=-1.1%, Limit of agreement=-8.1%-6.0%). In addition, the correlations between the manual adjustment by 3D Auto RV and the CMR ( r=0.63-0.72, all P<0.001) was lower than the correlations between the 3D Auto RV and the CMR ( r=0.76-0.82, all P<0.001) for RV volumes and RVEF.Finally, 3D Auto RV had a good reproducibility. Conclusions:The new fully 3D Auto RV quantification software underestimate RV volumes that less than the conventional semi-automated Tomtec. And the 3D Auto RV quantification software can accurately evaluate the RVEF in patients after HT with rapid analysis and higher reproducibility, which may also support the routine adoption of this method during follow-ups of HT patients in the daily clinical workflow.