1.Establishment and application of a method for detecting Toxoplasma gondii based on recombinant polymerase amplification technology
Shao-zheng SONG ; Le-ying GU ; Ying-chao WU ; Ya-qin MENG ; Kang-ying YU ; Xiao-hua HUANG
Chinese Journal of Zoonoses 2025;41(2):107-112
To establish a method for detecting Toxoplasma gondii based on recombinant polymerase amplification(RPA)technology and apply it to clinical sample validation of pet cats.Using the 529 repeat sequence of the Toxoplasma gondii gene as the target gene sequence,primers and probes were designed,and the Rep-529 recombinant plasmid was constructed as the standard.A fluorescent RPA reaction system was established.Dilute the plasmid standard 10 times to different concentrations as the detection template for sensitivity testing;Specific testing was conducted using genomic DNA from several parasitic spe-cies,including Toxoplasma gondii,Cryptosporidium,Neosporidium,Trichinella spiralis,Giardia flagellata,Babesia bo-vis and Theileria annulata as templates;Simultaneously,fluorescence RPA and RT-PCR were used to detect 52 positive and 40 negative cats clinical samples,and the coincidence rate of the detection results of the two methods were compared and ana-lyzed.The RPA reaction system was successfully established using PTRep recombinant plasmid as the standard,ToxD-F/ToxD-R as the primer,and RepD-P as the fluorescent probe.The reaction temperature was constant at 39 ℃,the reaction time was 30 minutes,and the detection sensitivity was 1 copy/μL.There is no significant cross reaction with parasites such as Cryptosporidium,Neosporidium,Trichinella spiralis,Giardia,Babesia bovis and Theileria annulata,and the specificity is good.A total of 92 clinical fecal samples from cats were tested,and the positive coincidence rate of fluorescence RPA detection method was higher than that of conventional RT-PCR method(98.08%vs.82.69%),and the difference of the positive rate was not statistically significant(X2=1.392,P>0.05).The fluorescence RPA detection method for Toxoplasma gondii suc-cessfully established in this study has the characteristics of being fast,sensitive,specific,accurate,and reliable.It can be used as a rapid clinical detection kit for Toxoplasma gondii in cats and other animals,providing new technical support for the subsequent epidemiological monitoring and precise clinical diagnosis of toxoplasmosis in cats,other animals,and humans in the future.
2.Application of parathyroid 99Tc m-MIBI SPECT/CT on hungry bone syndrome in patients with renal secondary hyperparathyroidism after parathyroidectomy
Zejun CHEN ; Bin ZHOU ; Qianhuan HUANG ; Qing SHAO ; Chaoqun WU ; Qin XUE
Chinese Journal of Nuclear Medicine and Molecular Imaging 2025;45(10):612-616
Objective:To investigate the relationship of the number and morphological of parathyroid glands on 99Tc m-methoxyisobutylisonitrile (MIBI) SPECT/CT with hungry bone syndrome (HBS) in patients with renal secondary hyperparathyroidism (SHPT) after parathyroidectomy (PTX). Methods:Eighty renal SHPT patients (46 males, 34 females; age (50.3±11.0) years) who underwent PTX between January 2018 and June 2024 were retrospectively analyzed in Affiliated Jiangyin Hospital of Nantong University. The number of parathyroid glands detected on SPECT/CT, diameter of parathyroid gland and its volume were recorded and measured. Patients were divided into HBS group and non-HBS group. Independent-sample t test, Mann-Whitney U test and χ2 test were used to compare differences of various indicators between those 2 groups. Multivariate logistic regression was used to analyze influencing factors of HBS. Results:HBS group included 45 patients and the rest 35 patients belonged to non-HBS group. Age ( t=-3.61, P=0.001), preoperative alkaline phosphatase (ALP) ( Z=-4.65, P<0.001), preoperative parathyroid hormone ( Z=-2.34, P=0.019) and preoperative serum corrected calcium ( t=-2.71, P=0.008) were all significantly different between HBS group and non-HBS group. Patients with the number of parathyroid glands≥4 detected by SPECT/CT were more in HBS group than those in non-HBS group (82.2%(37/45) vs 51.4%(18/35); χ2=8.87, P=0.003), and the total volume of parathyroid glands (2.56(1.93, 4.44) vs 2.00(1.18, 2.94)cm 3;Z=-2.25, P=0.024) and the maximum diameter of parathyroid glands ((17.71±3.78) vs (15.87±3.91) mm; t=2.14, P=0.036) were significantly different between those 2 groups. Logistic regression analysis showed that preoperative ALP (odds ratio ( OR)=1.008, 95% CI: 1.002-1.014, P=0.015), preoperative serum corrected calcium ( OR=0.017, 95% CI: 0.000-0.869, P=0.042) and the number of parathyroid gland≥4 detected by SPECT/CT ( OR=4.156, 95% CI: 1.038-16.642, P=0.044) were independent influencing factors for HBS. The sensitivity of the number of parathyroid glands≥4 detected by SPECT/CT for diagnosing HBS was 82.2%(37/45). Conclusion:The number of parathyroid glands≥4 detected by SPECT/CT is an independent influencing factor for HBS, with high diagnostic sensitivity for HBS, thus having good clinical value.
3.Quantification of Atmospheric Total Reactive Nitrogen Oxides by Thermal Decomposition-Broadband Cavity Enhanced Absorption Spectroscopy
Dou SHAO ; Min QIN ; Wu FANG ; Bao-Bin HAN ; Ke TANG ; Jian-Ye XIE ; Xia-Dan ZHAO ; Zhi-Tang LIAO ; En-Bo REN
Chinese Journal of Analytical Chemistry 2025;53(3):387-396
Nitrogen oxides(NOx=NO+NO2)are important precursors of ozone(O3),and NOx and its oxides together constitute reactive nitrogen oxides(NOy)in the atmosphere.A comprehensive understanding of the total NOy level in the atmosphere is of great significance for a deeper understanding of the atmospheric nitrogen cycle and oxidation,as well as for formulating strategies for air pollution prevention and control.In this work,a thermal decomposition-broadband cavity enhanced absorption spectroscopy(TD-BBCEAS)technique for online measurement of total NOy in the atmosphere was developed.With this method,the NOy was efficiently converted into NO2,and the total NOy concentration in the atmosphere was indirectly obtained by measuring NO2.Focusing on the key factors affecting the measurement of total NOy,the influence of NO titration efficiency and other NOy component TD efficiency on measurement accuracy was emphasized.By changing the oxygen(O2)flow rate through the mercury lamp to alter the O3 concentration for titrating NO,the conversion efficiency of NO was evaluated.At O2 flow rate of 6 mL/min,the conversion efficiency of NO was greater than 99%.TD efficiency testing and analysis on NO2,peroxyacetyl nitrate(PAN),nitric acid(HNO3),and nitrous acid(HONO),which account for a large proportion of atmospheric NOy components,was carried out using 680℃as the optimal TD temperature for efficient conversion of NOy.With NO and HONO sample gases as typical verification gases,the conversion efficiency of NOy and the accuracy of NOy measurement by TD-BBCEAS system were verified by switching the on and off modes of mercury lamp and TD device.At integration time of 60 s,the detection limit of the system for NOy was 2.83×1010 molecules/cm3(60 s,2σ).A comparative measurement of actual atmospheric NOy was conducted between the TD-BBCEAS system and the NOy analyzer.The observation results showed a correlation coefficient(R2)of 0.98 and a slope of 0.93,further verifying the feasibility and accuracy of applying the TD-BBCEAS system to measurement of total NOy.
4.Construction of entrustable professional activity indicators for non-anesthesiology residents during rotation in the department of anesthesiology
Xinxin SHAO ; Jianhong YE ; Xiaodan WU ; Lu YANG ; Qingchun LIANG ; Qin ZHOU
Chinese Journal of Medical Education Research 2025;24(9):1212-1217
Objective:To establish a framework of entrustable professional activity indicators for non-anesthesiology residents during rotation in the Department of Anesthesiology, standardize the training process, and provide an effective evaluation method.Methods:Based on literature review and discussions, a preliminary expert consultation questionnaire was developed. From July to November 2024, a modified Delphi method was adopted and relevant experts were invited to assess the importance, validity, and feasibility of each indicator using a 5-point Likert scale. The results were used to screen and refine the entrustable professional activity indicators for non-anesthesiology residents during rotation in the Department of Anesthesiology.Results:In the two rounds of expert consultation, the valid questionnaire return rate reached 100.00%, with an expert judgment basis coefficient of 0.73 and a familiarity coefficient of 0.90. Based on expert feedback, the final framework retained 3 first-level indicators and 12 second-level indicators. Modifications included renaming "Anesthesia Management" to "Anesthetic Drug Management", swapping the order of "Intraoperative Emergency Management" and "Airway Management", and removing the indicator "Resuscitation Assessment".Conclusions:This study preliminarily developed a framework of entrustable professional activity indicators for non-anesthesiology residents during rotation in the Department of Anesthesiology. The framework provides a simple and practical competency evaluation method for standardized training, which can enhance training quality and effectiveness.
5.Establishment and application of a method for detecting Toxoplasma gondii based on recombinant polymerase amplification technology
Shao-zheng SONG ; Le-ying GU ; Ying-chao WU ; Ya-qin MENG ; Kang-ying YU ; Xiao-hua HUANG
Chinese Journal of Zoonoses 2025;41(2):107-112
To establish a method for detecting Toxoplasma gondii based on recombinant polymerase amplification(RPA)technology and apply it to clinical sample validation of pet cats.Using the 529 repeat sequence of the Toxoplasma gondii gene as the target gene sequence,primers and probes were designed,and the Rep-529 recombinant plasmid was constructed as the standard.A fluorescent RPA reaction system was established.Dilute the plasmid standard 10 times to different concentrations as the detection template for sensitivity testing;Specific testing was conducted using genomic DNA from several parasitic spe-cies,including Toxoplasma gondii,Cryptosporidium,Neosporidium,Trichinella spiralis,Giardia flagellata,Babesia bo-vis and Theileria annulata as templates;Simultaneously,fluorescence RPA and RT-PCR were used to detect 52 positive and 40 negative cats clinical samples,and the coincidence rate of the detection results of the two methods were compared and ana-lyzed.The RPA reaction system was successfully established using PTRep recombinant plasmid as the standard,ToxD-F/ToxD-R as the primer,and RepD-P as the fluorescent probe.The reaction temperature was constant at 39 ℃,the reaction time was 30 minutes,and the detection sensitivity was 1 copy/μL.There is no significant cross reaction with parasites such as Cryptosporidium,Neosporidium,Trichinella spiralis,Giardia,Babesia bovis and Theileria annulata,and the specificity is good.A total of 92 clinical fecal samples from cats were tested,and the positive coincidence rate of fluorescence RPA detection method was higher than that of conventional RT-PCR method(98.08%vs.82.69%),and the difference of the positive rate was not statistically significant(X2=1.392,P>0.05).The fluorescence RPA detection method for Toxoplasma gondii suc-cessfully established in this study has the characteristics of being fast,sensitive,specific,accurate,and reliable.It can be used as a rapid clinical detection kit for Toxoplasma gondii in cats and other animals,providing new technical support for the subsequent epidemiological monitoring and precise clinical diagnosis of toxoplasmosis in cats,other animals,and humans in the future.
6.Construction of entrustable professional activity indicators for non-anesthesiology residents during rotation in the department of anesthesiology
Xinxin SHAO ; Jianhong YE ; Xiaodan WU ; Lu YANG ; Qingchun LIANG ; Qin ZHOU
Chinese Journal of Medical Education Research 2025;24(9):1212-1217
Objective:To establish a framework of entrustable professional activity indicators for non-anesthesiology residents during rotation in the Department of Anesthesiology, standardize the training process, and provide an effective evaluation method.Methods:Based on literature review and discussions, a preliminary expert consultation questionnaire was developed. From July to November 2024, a modified Delphi method was adopted and relevant experts were invited to assess the importance, validity, and feasibility of each indicator using a 5-point Likert scale. The results were used to screen and refine the entrustable professional activity indicators for non-anesthesiology residents during rotation in the Department of Anesthesiology.Results:In the two rounds of expert consultation, the valid questionnaire return rate reached 100.00%, with an expert judgment basis coefficient of 0.73 and a familiarity coefficient of 0.90. Based on expert feedback, the final framework retained 3 first-level indicators and 12 second-level indicators. Modifications included renaming "Anesthesia Management" to "Anesthetic Drug Management", swapping the order of "Intraoperative Emergency Management" and "Airway Management", and removing the indicator "Resuscitation Assessment".Conclusions:This study preliminarily developed a framework of entrustable professional activity indicators for non-anesthesiology residents during rotation in the Department of Anesthesiology. The framework provides a simple and practical competency evaluation method for standardized training, which can enhance training quality and effectiveness.
7.Application of parathyroid 99Tc m-MIBI SPECT/CT on hungry bone syndrome in patients with renal secondary hyperparathyroidism after parathyroidectomy
Zejun CHEN ; Bin ZHOU ; Qianhuan HUANG ; Qing SHAO ; Chaoqun WU ; Qin XUE
Chinese Journal of Nuclear Medicine and Molecular Imaging 2025;45(10):612-616
Objective:To investigate the relationship of the number and morphological of parathyroid glands on 99Tc m-methoxyisobutylisonitrile (MIBI) SPECT/CT with hungry bone syndrome (HBS) in patients with renal secondary hyperparathyroidism (SHPT) after parathyroidectomy (PTX). Methods:Eighty renal SHPT patients (46 males, 34 females; age (50.3±11.0) years) who underwent PTX between January 2018 and June 2024 were retrospectively analyzed in Affiliated Jiangyin Hospital of Nantong University. The number of parathyroid glands detected on SPECT/CT, diameter of parathyroid gland and its volume were recorded and measured. Patients were divided into HBS group and non-HBS group. Independent-sample t test, Mann-Whitney U test and χ2 test were used to compare differences of various indicators between those 2 groups. Multivariate logistic regression was used to analyze influencing factors of HBS. Results:HBS group included 45 patients and the rest 35 patients belonged to non-HBS group. Age ( t=-3.61, P=0.001), preoperative alkaline phosphatase (ALP) ( Z=-4.65, P<0.001), preoperative parathyroid hormone ( Z=-2.34, P=0.019) and preoperative serum corrected calcium ( t=-2.71, P=0.008) were all significantly different between HBS group and non-HBS group. Patients with the number of parathyroid glands≥4 detected by SPECT/CT were more in HBS group than those in non-HBS group (82.2%(37/45) vs 51.4%(18/35); χ2=8.87, P=0.003), and the total volume of parathyroid glands (2.56(1.93, 4.44) vs 2.00(1.18, 2.94)cm 3;Z=-2.25, P=0.024) and the maximum diameter of parathyroid glands ((17.71±3.78) vs (15.87±3.91) mm; t=2.14, P=0.036) were significantly different between those 2 groups. Logistic regression analysis showed that preoperative ALP (odds ratio ( OR)=1.008, 95% CI: 1.002-1.014, P=0.015), preoperative serum corrected calcium ( OR=0.017, 95% CI: 0.000-0.869, P=0.042) and the number of parathyroid gland≥4 detected by SPECT/CT ( OR=4.156, 95% CI: 1.038-16.642, P=0.044) were independent influencing factors for HBS. The sensitivity of the number of parathyroid glands≥4 detected by SPECT/CT for diagnosing HBS was 82.2%(37/45). Conclusion:The number of parathyroid glands≥4 detected by SPECT/CT is an independent influencing factor for HBS, with high diagnostic sensitivity for HBS, thus having good clinical value.
8.Effect of Collagen Peptides on Function of Mouse T Lymphocytes under Simulated Microgravity
Shao-Yan SI ; Ya-Ya QIN ; Ying-Ying WU ; Xiao-Yu MA ; Ying SHANG ; Shu-Jun SONG ; Yan-Chuan GUO
Journal of Experimental Hematology 2024;32(4):1258-1263
Objective:To understand the effect of collagen peptides on the function of mouse lymphocytes under simulated microgravity.Methods:The splenocytes of mice were isolated,and the rotary cell culture system was used to simulate the microgravity.The T lymphocytes were stimulated with mitotic agents,concanavalin A(ConA),and the cells were treated with different concentrations of collagen peptides.The proliferation of lymphocytes and the levels of cytokines in the supernatant were detected.Results:Simulated microgravity could inhibit the proliferation of spleen T lymphocytes and decrease the level of cytokines in the supernatant.Collagen peptides could promote the lymphocyte proliferation and cytokine production in cells cultured under simulated microgravity.Conclusion:Collagen peptides may attenuate the inhibitory effect of simulated microgravity on T lymphocytes by regulating the cell proliferation and the secretion of cytokines.
9.Comparison of Jinzhen oral liquid and ambroxol hydrochloride and clenbuterol hydrochloride oral solution in the treatment of acute bronchitis in children: A multicenter, non-inferiority, prospective, randomized controlled trial.
Qinhua FAN ; Chongming WU ; Yawei DU ; Boyang WANG ; Yanming XIE ; Zeling ZHANG ; Wenquan SU ; Zizhuo WANG ; Changchang XU ; Xueke LI ; Ying DING ; Xinjiang AN ; Jing CHEN ; Yunying XIAO ; Rong YU ; Nan LI ; Juan WANG ; Yiqun TENG ; Hongfen LV ; Nian YANG ; Yuling WEN ; Xiaoli HUANG ; Wei PAN ; Yufeng LIU ; Xueqin XI ; Qianye ZHAO ; Changshan LIU ; Jian XU ; Haitao ZHANG ; Lie ZHUO ; Qiangquan RONG ; Yu XIA ; Qin SHEN ; Shao LI ; Junhong WANG ; Shengxian WU
Acta Pharmaceutica Sinica B 2024;14(12):5186-5200
The comparison between traditional Chinese medicine Jinzhen oral liquid (JZOL) and Western medicine in treating children with acute bronchitis (AB) showed encouraging outcomes. This trial evaluated the efficacy and safety of the JZOL for improving cough and expectoration in children with AB. 480 children were randomly assigned to take JZOL or ambroxol hydrochloride and clenbuterol hydrochloride oral solution for 7 days. The primary outcome was time-to-cough resolution. The median time-to-cough resolution in both groups was 5.0 days and the antitussive onset median time was only 1 day. This randomized controlled trial showed that JZOL was not inferior to cough suppressant and phlegm resolving western medicine in treating cough and sputum and could comprehensively treat respiratory and systemic discomfort symptoms. Combined with clinical trials, the mechanism of JZOL against AB was uncovered by network target analysis, it was found that the pathways in TRP channels like IL-1β/IL1R/TRPV1/TRPA1, NGF/TrkA/TRPV1/TRPA1, and PGE2/EP/PKA/TRPV1/TRPA1 might play important roles. Animal experiments further confirmed that inflammation and the immune regulatory effect of JZOL in the treatment of AB were of vital importance and TRP channels were the key mechanism of action.
10.Vascular segmentation and reconstruction in diabetic retinopathy based on deep learning
Shiyi XU ; Minghui CHEN ; Yi SHAO ; Kaibo QIN ; Yuquan WU ; Zhijie YIN ; Zhengqi YANG
Chinese Journal of Medical Physics 2024;41(10):1256-1264
A method capable of retinal vessel segmentation and three-dimensional(3D)reconstruction is proposed for the early diagnosis of diabetic retinopathy.The 3D reconstruction can avoid the misjudgments of blood vessel length,curvature and branch angle after segmentation,which will affect the early diagnosis.IAAnet algorithm for retinal image segmentation combines traditional Unet with Inception V3,atrous spatial pyramid pooling and AttentionGates to reduce information loss and avoid over-fitting,thereby improving the network's ability to extract features.The projection reconstruction method is used to restore the 3D information of blood vessels,and supports the adjustments of brightness and contrast,so that doctors can better observe the real state of blood vessels.The proposed algorithm has an accuracy,recall rate,F1 score,intersection over union and area under ROC curve of 97.68%,96.07%,97.26%,92.79%and 94.00%,respectively.Compared with other networks,IAAnet algorithm exhibits higher segmentation accuracy,and can obtain more vascular information in 3D image after 3D projection reconstruction to assist in the early diagnosis.

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