Anxiety is a major emotional disorder manifested in the individual's expectation of future threats.The incidence rate of anxiety is about 7.3％,with the highest lifetime prevalence rate among mental health conditions.The mechanism of anxiety overlaps with depression,and anxiety is a typical symptom of various mental diseases or emotional disorders in traditional Chinese medicine.The high rates of comorbidity and disability pose serious threats to people's health.Animal models are important tools for studying anxiety and are of great use for deciphering the pathogenesis of anxiety and for developing drugs.The traditional paradigm of stress-induced anxiety,however,is relatively limited.Based on traditional theory combined with clinical and animal experimental data,we propose a new hypothesis of"insufficiency of ZhiYi'causing anxiety,defined as"an anxiety state induced by the inability of an individual to meet their own needs,limited or lacking after multiple attempts,rendered hindered and powerless by an inability to meet their desires".This hypothesis is more in line with the typical manifestations of despair,lack of pleasure,and social withdrawal in clinical patients,and is supported by traditional theory and experimental data showing"hunger but unable to eat,food but unable to obtain,and gain but not full".Based on this,the established modeling paradigm is easy to apply,with good repeatability and low cost,and can be used to establish anxiety models in rats and mice,to provide a theoretical and model basis for the development and pharmacological evaluation of anti-anxiety drugs.

This study investigated the expression of the soluble nontoxic mutant CRM197 of diphtheria toxin,to prepare a safe and effective recombinant CRM197 protein.Codon-optimized CRM197 gene sequences were cloned into the pCold Ⅱ and pET-28a(+)prokaryotic expression vectors.The successfully cloned recombinant plasmids were screened and transformed into Escherichia coli BL21(DE3)competent cells.After induction of expression,the protein expression type was determined through SDS-PAGE analysis.The conditions for expression of the solublerecombinant protein were then optimized.The recombinant CRM197 protein was purified through two rounds of Ni-NTA affinity chromatography.The purified protein was used to immunize mice,and antibody levels in the se-rum were measured with ELISA.The codon adaptation index(CAI)of the optimized sequence increased from 0.72 to 0.93.The recom-binant plasmids pET-28a(+)-CRM197 and pCold Ⅱ-CRM197 were successfully constructed,as confirmed through colony PCR and double digestion.Expression analysis revealed that pET-28a(+)-CRM197 was expressed primarily as inclusion bodies,whereas pColdⅡ-CRM197 was expressed predominantly in soluble form.The conditions for soluble protein expression via pCold Ⅱ-CRM197 were optimized.When the inoculum was 3%and the IPTG concentration was 0.4 mmol/L,induction at 20 ℃ for 24 h significantly increased the expression of the soluble target protein.The pCold Ⅱ-CRM197 recombinant protein was purified from the supernatant with Ni-NTA affinity chromatography,thus resulting in a target protein with a purity greater than 98%.ELISA after three rounds of immuniza-tion indicated that the levels of IgG,IgM,IgG1,and IgG2a antibodies in the serum in immunized mice were significantly higher than those in the control group.In summary,the CRM197 recombinant protein was successfully prepared with the pCold Ⅱvector and exhib-ited high soluble expression,high purity,and favorable immunogenicity.

Anxiety is a major emotional disorder manifested in the individual's expectation of future threats.The incidence rate of anxiety is about 7.3％,with the highest lifetime prevalence rate among mental health conditions.The mechanism of anxiety overlaps with depression,and anxiety is a typical symptom of various mental diseases or emotional disorders in traditional Chinese medicine.The high rates of comorbidity and disability pose serious threats to people's health.Animal models are important tools for studying anxiety and are of great use for deciphering the pathogenesis of anxiety and for developing drugs.The traditional paradigm of stress-induced anxiety,however,is relatively limited.Based on traditional theory combined with clinical and animal experimental data,we propose a new hypothesis of"insufficiency of ZhiYi'causing anxiety,defined as"an anxiety state induced by the inability of an individual to meet their own needs,limited or lacking after multiple attempts,rendered hindered and powerless by an inability to meet their desires".This hypothesis is more in line with the typical manifestations of despair,lack of pleasure,and social withdrawal in clinical patients,and is supported by traditional theory and experimental data showing"hunger but unable to eat,food but unable to obtain,and gain but not full".Based on this,the established modeling paradigm is easy to apply,with good repeatability and low cost,and can be used to establish anxiety models in rats and mice,to provide a theoretical and model basis for the development and pharmacological evaluation of anti-anxiety drugs.

This study investigated the expression of the soluble nontoxic mutant CRM197 of diphtheria toxin,to prepare a safe and effective recombinant CRM197 protein.Codon-optimized CRM197 gene sequences were cloned into the pCold Ⅱ and pET-28a(+)prokaryotic expression vectors.The successfully cloned recombinant plasmids were screened and transformed into Escherichia coli BL21(DE3)competent cells.After induction of expression,the protein expression type was determined through SDS-PAGE analysis.The conditions for expression of the solublerecombinant protein were then optimized.The recombinant CRM197 protein was purified through two rounds of Ni-NTA affinity chromatography.The purified protein was used to immunize mice,and antibody levels in the se-rum were measured with ELISA.The codon adaptation index(CAI)of the optimized sequence increased from 0.72 to 0.93.The recom-binant plasmids pET-28a(+)-CRM197 and pCold Ⅱ-CRM197 were successfully constructed,as confirmed through colony PCR and double digestion.Expression analysis revealed that pET-28a(+)-CRM197 was expressed primarily as inclusion bodies,whereas pColdⅡ-CRM197 was expressed predominantly in soluble form.The conditions for soluble protein expression via pCold Ⅱ-CRM197 were optimized.When the inoculum was 3%and the IPTG concentration was 0.4 mmol/L,induction at 20 ℃ for 24 h significantly increased the expression of the soluble target protein.The pCold Ⅱ-CRM197 recombinant protein was purified from the supernatant with Ni-NTA affinity chromatography,thus resulting in a target protein with a purity greater than 98%.ELISA after three rounds of immuniza-tion indicated that the levels of IgG,IgM,IgG1,and IgG2a antibodies in the serum in immunized mice were significantly higher than those in the control group.In summary,the CRM197 recombinant protein was successfully prepared with the pCold Ⅱvector and exhib-ited high soluble expression,high purity,and favorable immunogenicity.

OBJECTIVE: To investigate clinical effect of sternoclavicular hook plate in treating acute proximal clavicle fracture. METHODS: The clinical of 12 patients with acute unstable proximal clavicle fracture from June 2016 to June 2019 were retrospectively analyzed. There were 8 males and 4 females, aged from 46 to 63 years old. Ten patients caused by car accident and 2 patients caused by high falling. All patients had multiple injuries;the time from injury to surgery ranged from 2 to 14 d. All patients were treated with domestic sternoclavicular joint hook plate. The operative time ranged from 40 to 115 min. The intraoperative bleeding volume ranged from 30 to 110 ml, follow-up time ranged from 10 to 36 months, the fracture healing time ranged from 8 to 18 weeks. At the latest follow-up, the efficacy was evaluated by using shoulder joint function score (Rockwood score). RESULTS: All 12 patients were followed up, with no obvious pain at the latest follow-up. The rockwood scores of the affected shoulder ranged from 13 to 14, and the healthy shoulder ranged from 14 to 15. CONCLUSION The sternocleidoclavicular joint plate is fixed with preformed plate. The cantilever is designed to retain the motion of the sternoclavicular joint. It's safe and simple, avoid, the injury of important organs during operation, and has a good prognosis. It is an ideal fixation method for the treatment of proximal clavicle fracture.
Male ; Female ; Humans ; Middle Aged ; Clavicle/injuries* ; Sternoclavicular Joint/injuries* ; Retrospective Studies ; Fracture Fixation, Internal/methods* ; Treatment Outcome ; Fractures, Bone/surgery*

8-hydroxy-2'-deoxyguanosine (8-OHdG) is a sensitive and stable biomarker for evaluating DNA oxidative damage. A rapid and sensitive colloidal gold immunochromatographic strip was developed for 8-OHdG detection by a competitive method. The sample pad (glass cellulose film), bonding pad (glass cellulose film), nitrocellulose film and absorbent pad were pasted on the polyvinyl chloride (PVC) base plate to construct the test strip. Colloidal gold (AuNPs) was prepared by the reduction of chloroauric acid with sodium citrate. 8-OHdG antibody (Ab) was coated on the outer layer of AuNPs to form Ab@AuNPs as a probe. Bovine serum albumin (BSA) and 8-OHdG were conjugated with carbodiimide hydrochloride to prepare an artificial antigen, which was used as the coating antigen of detection line. Goat anti mouse polyclonal antibody IgG was used as the coating antibody of control line. The experimental parameters were optimized including the type of nitrocellulose membrane, the formula of loading solution, and the spraying amount of gold labeled antibody. The results showed that the appropriate nitrocellulose membrane was CN 95. The optimal loading solution included BSA (1%), Tween-20 (3%), sucrose (3%) and NaCl (0.9%). The optimal spraying amount of gold labeled antibody was 4 μL. 8-OHdG can be detected by the strip under visible light, and the level of 8-OHdG in urine can be preliminarily determined by comparing the color intensity of T line and C line. The 8-OHdG concentration in urine was further calculated by the gray value of T line and the threshold of detection was 2.55 μg/L. This colloidal gold immunochromatographic strip is simple, rapid and specific for detecting 8-OHdG in human urine to preliminarily evaluate the human status.
8-Hydroxy-2'-Deoxyguanosine ; Animals ; Antibodies, Monoclonal ; Gold ; Gold Colloid/chemistry* ; Metal Nanoparticles ; Mice ; Sensitivity and Specificity

Objective:To compare the prosthesis locations and postoperative hip functions between supercapsular percutaneously-assisted total hip (SuperPATH) approach and traditional posterolateral approach (PLA) in total hip arthroplasty.Methods:A retrospective analysis was conducted of the 107 patients who had undergone unilateral total hip arthroplasty at Department of Orthopedic Surgery, The First Affiliated Hospital to Soochow University from August 2016 to February 2019. They were divided into 2 groups according to their surgical approaches. In the SuperPATH group of 54 cases, there were 20 males and 34 females with an age of (64.3±9.1) years; in the PLA group of 53 cases, there were 20 males and 33 females with an age of (62.2±10.6) years. The 2 groups were compared in terms of abduction angle, ratio of abduction angle to safety zone, anteversion angle, ratio of anteversion angle to safety zone, retroversion angle, incidence of retroversion, and differences in eccentricity and lower limb length on the first day after operation, and Harris hip scores at 1 week, 3 months and the last follow-up postoperatively. Their complications were also recorded as well.Results:There were no statistically significant differences in the preoperative general data between the 2 groups, showing comparability ( P>0.05). The postoperative imaging data were complete for the 107 patients who had been followed up satisfactorily for 14 to 36 months (average, 25 months). The SuperPATH group had significantly larger retroversion angle (13.6°±9.6°) and incidence of retroversion (18.5%, 10/54), significantly smaller difference in eccentricity [0.26 (0.13,0.49) cm], and significantly higher Harris hip score [(74.8±7.8) points] at one week after surgery than those in the PLA group [3.0°±1.0°; 5.7%, 3/53; 0.38 (0.13,0.70) cm; (72.0±6.7) points] ( P<0.05). There were no statistically significant differences between the 2 groups in abduction angle, ratio of abduction angle to safety zone, anteversion angle, ratio of anteversion angle to safety zone, difference in lower limb length, or Harris hip scores at 3 months or the last follow-up postoperatively (all P>0.05). Follow-ups in both groups observed no more than one case of dislocation which responded to manual reduction. Conclusion:The minimally invasive SuperPATH approach may obtain better femoral eccentricity and higher early hip function scores than the traditional posterolateral approach, but may lead to a higher incidence of retroversion after prosthesis placement.

Objective:To explore the effect of the location and size of region of interest (ROI) on the measurement of liver fat by means of quantitative computed tomography (QCT).Methods:A total of 98 subjects who were examined with QCT for bone mineral density examination from December 25, 2019 to January 17, 2020 were recruited continuously from the Department of Health Management of Henan Provincial People′s Hospital. The liver fat content was measured by QCT workstation. The ROI was located respectively in the left lobe, the right anterior lobe and the right posterior lobe of the liver, and it was measured independently by the A measurer and B measurer. The central position of the ROI was fixed and the diameter was increased, and it was measured by the A measurer. In this study, Friedman test was used to compare the differences of measurement results in different positions or sizes of ROI, and intra-class correlation coefficients (ICC) was used to evaluate the repeatability of inter-measurers.Results:There was a significant difference for liver fat content under different positions of ROI (χ2=62.306, P<0.001), but no difference under different seizes of ROI (χ2=1.088, P=0.581). The ICC values of the inter-measurers repeatability analysis of the A measurer and B measurer in the left lobe, right anterior lobe and right posterior lobe of the liver were 0.847, 0.917 and 0.874, all more than 0.75, and the reproducibility was good. Conclusions:When QCT technique is applied to the measurement of liver fat content, the location conditions of ROI may affect results, so it is necessary to select multiple ROI in the whole liver for measurement. The inter-measurers repeatability of QCT in different parts of the liver is good.

Objective:To provide support for the clinical application of quantitative computed tomography (QCT) in the measurement of liver fat content, this study evaluated the intra-observer and inter-observer reproducibility of liver fat content measured by QCT in a population receiving physical examinations.Methods:From April to July 2019, 291 people were consecutively selected who underwent QCT examination in the health management department of Henan Provincial People′s Hospital. There were 214 males (73.5%) and 77 females (26.5%), aged 48.7±11.0. We measured liver fat content by QCT workstation. Three observers (A, B, C) measured their liver fat content independently, then observer A performed re-testing two weeks later. The mean value of the two measurements from observer A was taken as the final result. Measurement data were described by mean±SD. Intra-observer and inter-observer reproducibility were assessed using intra-class correlation coefficients ( ICC). Results:The first measurement result for observer A was 10.46±5.55 and the second measurement for observer A was 10.66±5.59, resulting in a final value of 10.56±5.51. The measurement results of observers B and C were 10.70±5.45 and 10.86±5.77, respectively. The ICC value of liver fat content values measured by the three observers was 0.960 (95% CI: 0.951-0.967, P<0.001) and the ICC value of liver fat content values for the two measurements of observer A was 0.953 (95% CI: 0.941-0.962, P<0.001). The ICC values were>0.75, so reproducibility of results was good. Conclusions:If the measurement method is consistent, the results for liver fat content measured by a conventional CT scanner and QCT workstation will have good reproducibility between and within observers, and will also have certain clinical application prospects.

Objective:To investigate the prevalence of dysphagia in old people in nursing homes in Ningbo and analyze its related factors. Methods:From August, 2017 to April, 2018, 997 people aged 60 or above were selected from eight nursing homes in four districts in Ningbo. They were investigated with general questionnaire, Kubota water swallowing test, body mass index (BMI) and Quality of Life Index (QLI), and the data were analyzed with Logistic regression. Results:There were 259 (25.98%) persons with dysphagia. Age, male, hypertension, diabetes, hyperlipidemia, coronary heart disease, stroke and fracture were risk factors for dysphagia (P < 0.01). There were 19.2% with low BMI in total, and 63.3% for dysphagia. The total score of QLI decreased with age (F = 18.706, P < 0.001), more in males than in females (t = 2.516, P = 0.012), and less in those with dysphagia than with normal swallowing (t = 7.176, P < 0.001). Conclusion:Dysphagia is prevalent in the old people in nursing homes in Ningbo, which needs early intervention.