1.Role of artificial intelligence in medical image analysis.
Lu WANG ; Shimin ZHANG ; Nan XU ; Qianqian HE ; Yuming ZHU ; Zhihui CHANG ; Yanan WU ; Huihan WANG ; Shouliang QI ; Lina ZHANG ; Yu SHI ; Xiujuan QU ; Xin ZHOU ; Jiangdian SONG
Chinese Medical Journal 2025;138(22):2879-2894
With the emergence of deep learning techniques based on convolutional neural networks, artificial intelligence (AI) has driven transformative developments in the field of medical image analysis. Recently, large language models (LLMs) such as ChatGPT have also started to achieve distinction in this domain. Increasing research shows the undeniable role of AI in reshaping various aspects of medical image analysis, including processes such as image enhancement, segmentation, detection in image preprocessing, and postprocessing related to medical diagnosis and prognosis in clinical settings. However, despite the significant progress in AI research, studies investigating the recent advances in AI technology in the aforementioned aspects, the changes in research hotspot trajectories, and the performance of studies in addressing key clinical challenges in this field are limited. This article provides an overview of recent advances in AI for medical image analysis and discusses the methodological profiles, advantages, disadvantages, and future trends of AI technologies.
Artificial Intelligence
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Humans
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Image Processing, Computer-Assisted/methods*
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Neural Networks, Computer
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Deep Learning
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Diagnostic Imaging/methods*
2.Application of delayed replantation of degloving skin preserved at 4 ℃ in treatment of limb degloving injuries.
Qianqian XU ; Jihai XU ; Yijun SHEN ; Chenxi ZHANG ; Hangchong SHEN ; Tianxiang HUANG ; Chenlin LU ; Xin WANG
Chinese Journal of Reparative and Reconstructive Surgery 2025;39(1):95-99
OBJECTIVE:
To investigate the effectiveness of delayed replantation of degloving skin preserved at 4℃ in treatment of limb degloving injuries.
METHODS:
Between October 2020 and October 2023, 12 patients with limb degloving injuries were admitted. All patients had severe associated injuries or poor wound conditions that prevented primary replantation. There were 7 males and 5 females; age ranged from 29 to 46 years, with an average of 39.2 years. The causes of injury included machine entanglement in 6 cases, traffic accidents in 5 cases, and sharp instrument cuts in 1 case. Time from injury to hospital admission was 0.5-3.0 hours, with an average of 1.3 hours. Injury sites included upper limbs in 7 cases and lower limbs in 5 cases. The range of degloving skin was from 5 cm×4 cm to 15 cm×8 cm, and all degloving skins were intact. The degloving skin was preserved at 4℃. After the patient's vital signs became stable and the wound conditions improved, it was trimmed into medium-thickness skin grafts for replantation. The degloving skin was preserved for 3 to 7 days. At 4 weeks after replantation, the viability of the degloving skin grafts was assessed, including color, elasticity, and sensation of pain. The Vancouver Scar Scale (VSS) was used to assess the scars of the skin grafts during follow-up.
RESULTS:
At 4 weeks after replantation, 8 cases of skin grafts completely survived and the color was similar with normal skin, with a survival rate of 66.67%. The elasticity of skin grafts (R0 value) ranged from 0.09 to 0.85, with an average of 0.55; moderate pain was reported in 4 cases, mild pain in 3 cases, and no pain in 5 cases. All patients were followed up 12 months. Over time, the VSS scores of all 12 patients gradually decreased, with a range of 4-11 at 12 months (mean, 6.8).
CONCLUSION
For limb degloving injuries that cannot be replanted immediately and do not have the conditions for deep low-temperature freezing preservation, the method of preserving the degloving skin at 4℃ for delayed replantation can be chosen.
Humans
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Male
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Adult
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Replantation/methods*
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Female
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Degloving Injuries/surgery*
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Middle Aged
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Skin Transplantation/methods*
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Treatment Outcome
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Extremities/injuries*
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Time Factors
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Skin/injuries*
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Tissue Preservation/methods*
3.Guijianyu alleviates advanced glycation endproducts-induced mouse renal podocyte injury by inhibiting the AGEs-RAGE signaling pathway.
Qianqian MA ; Yuqi NIU ; Mingyu ZUO ; Xin LI ; Junke FU ; Jinjin WANG
Journal of Southern Medical University 2025;45(9):1938-1945
OBJECTIVES:
To investigate the mechanism by which Guijianyu ameliorates podocyte injury in a mouse model of diabetic kidney disease (DKD) induced by advanced glycation endproducts (AGEs).
METHODS:
Sixty db/db mouse models of DKD were randomized equally into 5 groups for treatment with saline, Guijianyu extract at 3 doses or irbesartan for 12 weeks, and the changes in renal pathology and structure were observed using transmission electron microscopy, and the expressions of related genes and key proteins were detected using RT-qPCR and immunohistochemistry. In cultured MPC-5 cells incubated with 50 mg/L AGEs-BSA for 24 h, the effect of different concentrations of Guijianyu extract on cell viability was examined with CCK-8 assay; Western blotting was performed to detect the protein expressions of RAGE, VEGFA, TNF-α, NF-κB(p65), IL-6 and caspase-3, and the mRNA expressions of RAGE, NF-κB (p65), VEGFA and IL-6 were detected with RT-qPCR.
RESULTS:
In mouse models of DKD, treatment with high-dose Guijianyu extract significantly reduced renal expressions of RAGE, VEGFA, NF-κB(p65), and IL-6 proteins and the mRNA expressions of RAGE, NF-κB, and IL-6. In MPC-5 cells, exposure to AGEs significantly reduced cell viability and increased the protein expressions of RAGE, NF‑κB (p65), VEGFA, TNF-α, IL-6 and caspase-3 (P<0.05) and mRNA expressions of RAGE, NF-κB (p65), VEGFA, and IL-6. Treatment with Guijianyu extract obviously improved cell viability and reduced the expressions of RAGE, NF-κB(p65), VEGFA, TNF-α, IL-6, and caspase-3. Furthermore, Guijianyu extract effectively reversed RAGE agonist-induced elevation of protein expressions of RAGE, VEGFA, TNF-α, IL-6, and caspase-3 and mRNA expressions of RAGE, NF-κB (p65), IL-6, and VEGFA in MPC-5 cells.
CONCLUSIONS
Guijianyu extract ameliorates AGEs-induced mouse renal podocyte injury in DKD by inhibiting the activation of AGEs-RAGE signaling pathway and reducing the expressions of pro-inflammatory cytokines and vascular endothelial growth factors.
Animals
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Glycation End Products, Advanced
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Drugs, Chinese Herbal/pharmacology*
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Mice
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Signal Transduction/drug effects*
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Podocytes/pathology*
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Diabetic Nephropathies/drug therapy*
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Receptor for Advanced Glycation End Products
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Vascular Endothelial Growth Factor A/metabolism*
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Interleukin-6/metabolism*
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Male
4.Comparison of Effect of Hirudo, Notoginseng Radix et Rhizoma, and Their Combinations on Renal Fibrosis in Rats with Chronic Renal Failure
Munan WANG ; Xuekuan HUANG ; Hongyu LUO ; Ke XU ; Xin CHEN ; Ling ZUO ; Qianqian SONG ; Peng DENG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(2):110-117
ObjectiveTo observe the effects of Hirudo, Notoginseng Radix et Rhizoma, and drug pair on renal pathological morphology and protein phosphatase 2A (PP2A)/adenylate activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signal pathway in rats with chronic renal failure (CRF). MethodThe 55 male SD rats were randomly divided into a normal group (n=11) and a modeling group (n=44). The normal group was fed conventionally, and the modeling group was given 0.25 g·kg-1·d-1 adenine by gavage for 28 days to replicate the CRF model. After successful modeling, rats were randomly divided into model group, Hirudo group (3 g·kg-1·d-1), Notoginseng Radix et Rhizoma group (3 g·kg-1·d-1), and Hirudo + Notoginseng Radix et Rhizoma group (3 g·kg-1·d-1), with 9 rats in each group. The normal group and model group were given a constant volume of normal saline by intragastric administration for 30 days. At the end of the experiment, the levels of serum creatinine (SCr) and urea nitrogen (BUN) in all groups were measured. The renal pathological morphology changes were observed by hematoxylin-eosin (HE) staining, Masson staining, and electron microscopy. The mRNA expressions of PP2A, AMPK, and mTOR were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). The protein expression levels of PP2A, AMPK, phosphorylation(p)-AMPK, mTOR, and p-mTOR in renal tissue were detected by Western blot. ResultCompared with the normal group, the renal pathological structure changes were obvious, and the levels of SCr and BUN were significantly increased. The mRNA expression of PP2A, protein expression of PP2A, and p-mTOR/mTOR expression were significantly increased, and the p-AMPK/AMPK was significantly decreased in the model group (P<0.05). Compared with the model group, the renal pathological morphology changes were significantly improved, and the levels of SCr and BUN were significantly decreased. The mRNA expression of PP2A, protein expression of PP2A, and p-mTOR/mTOR expression in the renal tissue were significantly decreased, and the p-AMPK/AMPK was significantly increased (P<0.05) in all groups after drug intervention. In addition, the effect in the Hirudo+Notoginseng Radix et Rhizoma group was better. The mRNA expression levels of AMPK and mTOR in the renal tissue were not significantly different among the normal group, model group, and other groups. ConclusionThe efficacy of Hirudo and Notoginseng Radix et Rhizoma pairs in improving renal fibrosis in rats with CRF is significantly better than that of the single drug, and its improvement on renal fibrosis in rats with CRF may be related to the regulation of PP2A/AMPK/mTOR signaling pathway.
5.Analysis of allergen-specific IgE in children with atopic dermatitis from 2021 to 2023 in a hospital of pediatric in Tianjin City
Ren NA ; Yuanmin SUN ; Ke LI ; Qinfeng LI ; Ying WANG ; Jiayi ZHANG ; Lisheng ZHENG ; Chaomeng FAN ; Qianqian XIN ; Xiaohui YANG ; Yongming SHEN
Chinese Journal of Preventive Medicine 2024;58(6):799-805
Objective:To explore the distribution of allergen-specific IgE (sIgE) for children with atopic dermatitis in Tianjin City and provide the evidences of clinical diagnosis and treatment.Methods:A retrospective cross-sectional study was conducted to analyze the children who were suspected of atopic dermatitis and tested for serum sIgE in the Tianjin Children′s Hospital from March 2021 to February 2023. Using first detection results only, a total of 1 841 serum samples were tested for twenty common allergens. The method was the enzyme-linked immune capture assay. The allergen epidemiological characteristics were statistically analyzed by Chi square test based on the children′s characteristics and factors such as different sexes, ages and seasons by the mass data.Results:Among the 1 841 cases, the results showed that 1 247 (67.73%) were sensitized to at least 1 allergen-sIgE, comprising to 49.86% (918/1 841) to food allergen-sIgE and 47.96% (883/1 841) to aeroallergen-sIgE. The top three food allergens-sIgE were egg 32.10% (591/1 841), milk 25.91% (477/1 841) and wheat flour 14.61% (269/1 841); the top three positive rates of aeroallergens-sIgE were house dust 24.33% (448/1 841), alternaria 20.59% (379/1 841) and dermatophagoides farinae 14.83% (273/1 841). The positive rates of food allergens-sIgE were the highest in the 1-3 years old group (64.11%, 434/677) ( χ2=122.854, P<0.001), while the positive rates of aeroallergens-sIgE were higher in the 11-14 years old group (71.26%, 62/87) ( χ2=134.968, P<0.001). No seasonal difference was revealed in the overall positive rate of food allergen-sIgE and aeroallergen-sIgE ( χ2=4.047, P=0.256; χ2=7.549, P=0.056). The positive rates of soybean-sIgE and milk-sIgE were the highest in summer ( χ2=11.329, P=0.010; χ2=28.720 , P<0.001), whereas alternaria-sIgE and mugwort-sIgE were the highest in summer and autumn, respectively ( χ2=8.462, P=0.037; χ2=10.641 , P=0.014). Among the 1 841 cases, 32.21% were sensitized to three or more allergens-sIgE. The sIgE concentration levels of egg, milk and house dust were mainly level 1 to 2, and the proportions of level 3 and above were all under 15%; although the positive rates of crab, shrimp, and peanut were low, the proportions of grade 3 and above were all beyond 30%. Children sensitized to alternaria, dermatophagoides farinae, mugwort, and cat dander had higher sIgE concentration levels, which were 68.07%, 49.45%, 56.57% and 47.83% respectively. Conclusions:This study can reflect the epidemic characteristics of allergen-sIgE in children with atopic dermatitis in Tianjin region to a certain extent. Allergen-sIgE positivity in patients differed by age, and there were seasonal differences and grade distribution differences in the positive rates of some allergens-sIgE. It is necessary to reasonably avoid the high-risk allergens according to the epidemiological characteristics and clinical symptoms, which provide valuable information for the prevention, diagnosis and treatment of atopic dermatitis.
6.Analysis of allergen-specific IgE in children with atopic dermatitis from 2021 to 2023 in a hospital of pediatric in Tianjin City
Ren NA ; Yuanmin SUN ; Ke LI ; Qinfeng LI ; Ying WANG ; Jiayi ZHANG ; Lisheng ZHENG ; Chaomeng FAN ; Qianqian XIN ; Xiaohui YANG ; Yongming SHEN
Chinese Journal of Preventive Medicine 2024;58(6):799-805
Objective:To explore the distribution of allergen-specific IgE (sIgE) for children with atopic dermatitis in Tianjin City and provide the evidences of clinical diagnosis and treatment.Methods:A retrospective cross-sectional study was conducted to analyze the children who were suspected of atopic dermatitis and tested for serum sIgE in the Tianjin Children′s Hospital from March 2021 to February 2023. Using first detection results only, a total of 1 841 serum samples were tested for twenty common allergens. The method was the enzyme-linked immune capture assay. The allergen epidemiological characteristics were statistically analyzed by Chi square test based on the children′s characteristics and factors such as different sexes, ages and seasons by the mass data.Results:Among the 1 841 cases, the results showed that 1 247 (67.73%) were sensitized to at least 1 allergen-sIgE, comprising to 49.86% (918/1 841) to food allergen-sIgE and 47.96% (883/1 841) to aeroallergen-sIgE. The top three food allergens-sIgE were egg 32.10% (591/1 841), milk 25.91% (477/1 841) and wheat flour 14.61% (269/1 841); the top three positive rates of aeroallergens-sIgE were house dust 24.33% (448/1 841), alternaria 20.59% (379/1 841) and dermatophagoides farinae 14.83% (273/1 841). The positive rates of food allergens-sIgE were the highest in the 1-3 years old group (64.11%, 434/677) ( χ2=122.854, P<0.001), while the positive rates of aeroallergens-sIgE were higher in the 11-14 years old group (71.26%, 62/87) ( χ2=134.968, P<0.001). No seasonal difference was revealed in the overall positive rate of food allergen-sIgE and aeroallergen-sIgE ( χ2=4.047, P=0.256; χ2=7.549, P=0.056). The positive rates of soybean-sIgE and milk-sIgE were the highest in summer ( χ2=11.329, P=0.010; χ2=28.720 , P<0.001), whereas alternaria-sIgE and mugwort-sIgE were the highest in summer and autumn, respectively ( χ2=8.462, P=0.037; χ2=10.641 , P=0.014). Among the 1 841 cases, 32.21% were sensitized to three or more allergens-sIgE. The sIgE concentration levels of egg, milk and house dust were mainly level 1 to 2, and the proportions of level 3 and above were all under 15%; although the positive rates of crab, shrimp, and peanut were low, the proportions of grade 3 and above were all beyond 30%. Children sensitized to alternaria, dermatophagoides farinae, mugwort, and cat dander had higher sIgE concentration levels, which were 68.07%, 49.45%, 56.57% and 47.83% respectively. Conclusions:This study can reflect the epidemic characteristics of allergen-sIgE in children with atopic dermatitis in Tianjin region to a certain extent. Allergen-sIgE positivity in patients differed by age, and there were seasonal differences and grade distribution differences in the positive rates of some allergens-sIgE. It is necessary to reasonably avoid the high-risk allergens according to the epidemiological characteristics and clinical symptoms, which provide valuable information for the prevention, diagnosis and treatment of atopic dermatitis.
7.Analysis of peripheral retinal defocus in children and adolescents with low to moderate myopia and its influencing factors
Shoukuan ZHANG ; Qianqian LIU ; Qingsheng PENG ; Honglian GAO ; Xin SUN ; Jun JIANG ; Lei ZHANG
Recent Advances in Ophthalmology 2024;44(10):808-813
Objective To investigate the peripheral retinal defocus and its influencing factors in children and adoles-cents with low to moderate myopia.Methods Totally 281 children and adolescents aged 6-15 years were included in the study,and only the right eye was selected.After cycloplegic refraction as well as axial length(AL)and average corneal curvature(AveK)measurements,the patients were divided into low myopia(LM)group(-3.00 D≤SE≤-0.50 D)and moderate myopia(MM)group(-6.00 D≤SE<-3.00D)according to spherical equivalent(SE),and stratified compari-sons were made according to AL[AL1 group(23.00 mm≤AL≤24.00 mm),AL2 group(24.00 mm<AL≤25.00 mm),and AL3 group(25.00 mm<AL≤26.00 mm)]and AveK[AveK1 group(40.00 D≤ Ave K≤43.00 D)and AveK2 group(43.00 D<AveK≤46.00 D)].Multispectral refraction tomography was used to measure the refraction difference value(RDV),in-cluding TRDV(0° to 53°),RDV-15(0° to 15°),RDV-30(0° to 30°),RDV-45(0° to 45°),RDV-15-30(15° to 30°),RDV-30-45(30° to 45°),RDV-45-53(45° to 53°),RDV-S(superior),RDV-I(inferior),RDV-T(temporal)and RDV-N(na-sal).The RDV was compared in the groups divided according to SE,AL and AveK individually,and the correlation be-tween RDV and age,SE,AL and AveK was analyzed.Moreover,the factors affecting RDV in all ranges were analyzed by multiple linear regression.Results Compared with the LM group,the MM group had significant increases in TRDV,RDV-30,RDV-45,RDV-15-30,RDV-30-45,RDV-45-53,RDV-S,RDV-I and RDV-N(all P<0.05)and no significant differ-ence in RDV-15 and RDV-T(both P>0.05).According to the comparisons of AL groups and AveK groups,the TRDV,RDV-30,RDV-45,RDV-15-30,RDV-30-45,RDV-45-53,RDV-S,RDV-I and RDV-N in the AL2 group were significantly higher than those in the AL1 group(all P<0.05);the TRDV,RDV-30,RDV-45,RDV-15-30,RDV-30-45,RDV-45-53,RDV-S and RDV-N in the AL3 group were significantly higher than those in AL2 and AL1 groups,and RDV-I and RDV-T in the AL3 group were significantly higher than those in the AL1 group(both P<0.05);the TRDV,RDV-30,RDV-45,RDV-15-30,RDV-30-45,RDV-45-53,RDV-S,and RDV-I in the Ave K1 group were significantly higher than those in the AveK2 group(all P<0.05).The correlation analysis showed that TRDV,RDV-45,RDV-30-45,RDV-45-53,RDV-S and RDV-N were positively correlated with age and AL and negatively correlated with SE and Ave K;RDV-30,RDV-15-30 and RDV-I were positively cor-related with AL and negatively correlated with SE and AveK;RDV-T was positively correlated only with AL;RDV-15 was not correlated with age,SE,AL and AveK.Multiple linear regression analysis showed that age was the influencing factor of RDV-45-53 and RDV-S;AL was the influencing factor of TRDV,RDV-30,RDV-45,RDV-15-30,RDV-30-45,RDV-45-53,RDV-S and RDV-T;AveK was the influential factor of RDV-I;SE had no significant effect on RDV in all ranges.Conclu-sion Peripheral retinal defocus in children and adolescents with low to moderate myopia has reached hyperopic defocus,and hyperopic defocus is the least in patients with relatively short AL.Age,AL and AveK can affect peripheral retinal defo-cus in children and adolescents with low to moderate myopia,among which AL is the most important influencing factor.
8.Recommendations for prescription review of commonly used anti-seizure medications in treatment of children with epilepsy
Qianqian QIN ; Qian DING ; Xiaoling LIU ; Heping CAI ; Zebin CHEN ; Lina HAO ; Liang HUANG ; Yuntao JIA ; Lingyan JIAN ; Zhong LI ; Hua LIANG ; Maochang LIU ; Qinghong LU ; Xiaolan MO ; Jing MIAO ; Yanli REN ; Huajun SUN ; Yanyan SUN ; Jing XU ; Meixing YAN ; Li YANG ; Shengnan ZHANG ; Shunguo ZHANG ; Xin ZHAO ; Jie DENG ; Fang FANG ; Li GAO ; Hong HAN ; Shaoping HUANG ; Li JIANG ; Baomin LI ; Jianmin LIANG ; Jianxiang LIAO ; Zhisheng LIU ; Rong LUO ; Jing PENG ; Dan SUN ; Hua WANG ; Ye WU ; Jian YANG ; Yuqin ZHANG ; Jianmin ZHONG ; Shuizhen ZHOU ; Liping ZOU ; Yuwu JIANG ; Xiaoling WANG
Chinese Journal of Applied Clinical Pediatrics 2023;38(10):740-748
Anti-seizure medications (ASMs) are the main therapy for epilepsy.There are many kinds of ASMs with complex mechanism of action, so it is difficult for pharmacists to examine prescriptions.This paper put forward some suggestions on the indications, dosage forms/routes of administration, appropriateness of usage and dosage, combined medication and drug interaction, long-term prescription review, individual differences in pathophysiology of children, and drug selection when complicated with common epilepsy, for the reference of doctors and pharmacists.
9.Correlation between intestinal flora characteristics and hepatic and renal impairments in HIV-infected heterosexual men
Mingjun MA ; Jialu HUANG ; Lidan ZHU ; Panpan CHEN ; Yong ZHANG ; Haoran ZHANG ; Qiuli XU ; Qianqian LIU ; Shaotan XIAO ; Xin XIN
Shanghai Journal of Preventive Medicine 2023;35(12):1192-1198
ObjectiveTo explore the relationship between the intestinal flora and the impairment of liver and kidney in HIV-infected men who have heterosexual sex with healthy women. MethodsFecal samples from 41 HIV-infected heterosexual men who have sex with women (PMSW) and 43 age- and BMI-matched healthy heterosexual men who have sex with women (NMSW) were collected and subjected to 16S rDNA sequencing. The blood levels of AST, ALT, TBIL, UREA, Cr, UA, β2-MG and other liver and kidney function indicators were measured. Bioinformatics methods were used to analyze the characteristics of the intestinal flora of the patients in these two groups, to compare the differential bacteria strains, and to analyze their correlation with liver and kidney function indicators. ResultsIn comparison with NMSW, the alpha diversity of intestinal flora was decreased in PMSW, and the beta diversity analysis showed significant differences in flora characteristics between the two groups (P<0.05). The abundance of Clostridium, Phylum thick-walled, Trichosporon, and Clostridium tumefaciens decreased but Fusobacteriota increased (LDA score >4). The comparison of liver and kidney function indexes revealed that AST, β2-MG levels were higher in PMSW than in NMSW, while TBIL was lower in PMSW than in NMSW. The number of patients with abnormal β2-MG was much higher in PMSW than in NMSW, and the difference was statistically significant (P<0.001). It was also found that AST was negatively correlated with Clostridium (P<0.05); TBIL was negatively correlated with Clostridium and positively correlated with Phylum thick-walled and Trichosporon (P<0.05). β2-MG was negatively correlated with Phylum thick-walled, Clostridium, Trichosporon and Rumenococcus (P<0.05) and positively correlated with Clostridium (P<0.05). ConclusionIn PMSW group, the alpha diversity of the flora is decreased. AST and β2-MG levels are increased, and TBIL level is decreased. These changes were significantly correlated with different strains of bacteria in the intestinal flora.
10.Preparetion of anti-CD70 nanobody immunotoxin fused with PE38 and its killing effect on renal clear cell carcinoma 786-O cells
XU Xinlan ; LIU Chang ; ZHANG Xin ; HU Qianqian ; LI Jiangwei
Chinese Journal of Cancer Biotherapy 2023;30(8):665-671
[摘 要] 目的:构建靶向CD70分子的重组免疫毒素,通过表达、纯化制备PE38与抗CD70纳米抗体重组蛋白,体外抗肿瘤实验探究重组蛋白是否对高表达CD70分子的阳性肿瘤细胞具有杀伤活性。方法:通过基因工程手段,将CD70纳米抗体Nb 2B3基因片段通过一个连接子与pET21a-PE38基因片段相连,获得重组表达载体pET21a-Nb 2B3-PE38并转入BL21(DE3)感受态细胞中进行表达、纯化与鉴定。用间接ELISA及FACS法检测Nb 2B3-PE38与CD70分子的结合活性,MTT法检测Nb 2B3-PE38对高表达CD70分子的肾透明细胞癌786-O细胞的体外杀伤活性,Annexin Ⅴ-FITC/PI双染法检测Nb 2B3-PE38对786-O细胞凋亡的影响。结果:成功构建抗CD70纳米抗体重组免疫毒素Nb 2B3-PE38,纯化获得纯度>90%的重组蛋白,SDS-PAGE及WB检测结果表明目的蛋白正确表达,分子量为56 000。纯化后的Nb 2B3-PE38能与重组CD70抗原及786-O细胞表面的CD70分子特异性结合;25 µg/mL Nb 2B3-PE38即对786-O细胞产生极显著的杀伤作用(P<0.001),并且促进786-O细胞的细胞凋亡(P<0.01),其杀伤效应强于阳性对照顺铂(P<0.01)。结论:成功制备了特异性靶向CD70分子的免疫毒素Nb 2B3-PE38,其能够有效杀伤786-O细胞并诱导细胞凋亡且效果强于顺铂。

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