1.Preliminary study on the biological characteristics of heat shock cognate protein 20 of Schistosoma japonicum
Xingang YU ; Kaijian YUAN ; Yilong LI ; Xuanru MU ; Hui XU ; Qiaoyu LI ; Wenjing ZENG ; Zhiqiang FU ; Yang HONG
Chinese Journal of Schistosomiasis Control 2025;37(3):294-303
Objective To clone and express the heat shock cognate protein 20 (SjHsc20) of Schistosoma japonicum, and to preliminarily investigate its biological characteristics. Methods The target fragment of the SjHsc20 gene was amplified using PCR assay and cloned into the pET-28a(+) expression plasmid to generate the recombinant expression vector pET-28a(+)-SjH-sc20, which was then transformed into Escherichia coli BL21 (DE3) competent cells. The recombinant SjHsc20 (rSjHsc20) protein was induced with isopropyl β-D-thiogalactopyranoside (IPTG) and purified, and the expression of the rSjHsc20 protein was checked with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The immunogenicity of the rSjHsc20 protein was detected using Western blotting, and the transcriptional levels of SjHsc20 were quantified in S. japonicum worms at different developmental stages and in male and female adult worms using real-time quantitative PCR (RT-qPCR) assay. Thirty female BALB/c mice at ages 6 to 8 weeks were divided into three groups, including the rSjHsc20 immunization group, the PBS control group, and the ISA 206 adjuvant group, of 10 mice in each group. Mice in the rSjHsc20 immunization group were subcutaneously immunized with 20 μg rSjHsc20 on days 1, 15 and 31, and animals in the PBS control group were subcutaneously injected with the same volume of PBS on days 1, 15 and 31, while mice in the ISA 206 adjuvant group were subcutaneously immunized with the same volume of ISA 206 adjuvant on days 1, 15 and 31, respectively. All mice in each group were infected with (40 ± 2) S. japonicum cercariae via the abdomen 14 day following the last immunization. Levels of serum specific IgG and its subtypes IgG1 and IgG2 antibodies against rSjHsc20, and the serum titers of anti-rSjHsc20 antibody were detected in mice using indirect enzyme-linked immunosorbent assay (ELISA). All mice were sacrifice 42 days post-infection, and S. japonicum worms were collected from the hepatic portal vein and counted. The eggs per gram (EPG), worm burden reductions and egg burden reductions were estimated to evaluate the protective efficacy of the rSjHsc20 protein. Results The SjHsc20 gene had an open reading frame (ORF) with 756 bp in length and encoded 252 amino acids, and the rSjHsc20 protein had a relative molecular mass of approximately 29 kDa. The rSjHsc20 protein was recognized by the serum of mice infected with S. japonicum and the serum of mice immunized with the rSjHsc20 protein, indicating that rSjHsc20 had a good immunogenicity. There was a significant difference in the transcriptional levels of the SjHsc20 gene among the 7-day (1.001 4 ± 0.065 7), 12-day (2.268 3 ± 0.129 2), 21-day (1.378 5 ± 0.160 4), 28-day (1.196 4 ± 0.244 0), 35-day (1.646 3 ± 0.226 1), 42-day worms of S. japonicum (1.758 0 ± 0.611 1) (F = 38.45, P < 0.000 1), and the transcriptional level of the SjHsc20 gene was higher in the 12-day worms than in worms at other developmental stages (all P values < 0.000 1). The serum levels of anti-rSjHsc20 IgG antibody were 0.106 6 ± 0.010 7, 0.108 3 ± 0.010 4, and 0.553 2 ± 0.069 1 in the PBS control group, ISA 206 adjuvant group, and rSjHsc20 immunization group following the last immunization, respectively, and the serum levels of IgG1 antibody were 0.137 3 ± 0.054 0, 0.181 1 ± 0.096 8, and 1.765 8 ± 0.221 1, while the levels of IgG2a antibody were 0.280 3 ± 0.197 6, 0.274 0 ± 0.146 3, and 1.560 4 ± 0.106 0, respectively. There were significant differences in the serum levels of anti-rSjHsc20 IgG (F = 397.70, P < 0.000 1), IgG1 (F = 401.00, P < 0.000 1) and IgG2a antibodies (F = 229.70, P < 0.000 1) among the three groups, and the serum levels of anti-rSjHsc20 IgG, IgG1 and IgG2a antibodies were higher in the rSjHsc20 immunization group than in the PBS control group and the ISA 206 adjuvant group (all P values < 0.000 1). There was a significant difference in the IgG1/IgG2a ratio among the rSjHsc20 immunization group (1.177 2 ± 0.143 6), the PBS control group (0.428 4 ± 0.199 8) and the ISA 206 adjuvant group (0.559 9 ± 0.181 1) (F = 43.97, P < 0.000 1), and the IgG1/IgG2a ratio was > 1 in the rSjHsc20 immunization group, which was higher than in the PBS control group and the ISA 206 adjuvant group (both P values < 0.000 1). The titers of serum anti-rSjHsc20 antibody were all above 1∶16 384 in the rSjHsc20 immunization group following immunizations on days 1, 15 and 31, indicating that the rSjHsc20 protein had a strong immunogenicity. The mean worm burdens were (16.60±5.75), (15.80±5.58) worms per mouse and (14.40±5.75) worms per mouse in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group 42 days post-infection with S. japonicum cercariae (F = 0.50, P > 0.05), and the EPG were 68 370 ± 22 690, 67 972 ± 19 502, and 41 075 ± 13 251 in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group (F = 4.55, P < 0.05), with lower EPG in the PBS control group and the ISA 206 adjuvant group than in the rSjHsc20 immunization group (both P values < 0.05). Immunization with the rSjHsc20 protein resulted in a worm burden reduction of 13.25% and an egg burden reduction of 39.92% relative to the PBS control group. Conclusions SjHsc20 is successfully cloned and expressed, and the rSjHsc20 protein induces partial immunoprotective effects in mice, which provides a basis for deciphering the biological functions of SjHsc20 and assessing the potential of SjH-sc20 as a vaccine candidate.
2.Gallstones, cholecystectomy, and cancer risk: an observational and Mendelian randomization study.
Yuanyue ZHU ; Linhui SHEN ; Yanan HUO ; Qin WAN ; Yingfen QIN ; Ruying HU ; Lixin SHI ; Qing SU ; Xuefeng YU ; Li YAN ; Guijun QIN ; Xulei TANG ; Gang CHEN ; Yu XU ; Tiange WANG ; Zhiyun ZHAO ; Zhengnan GAO ; Guixia WANG ; Feixia SHEN ; Xuejiang GU ; Zuojie LUO ; Li CHEN ; Qiang LI ; Zhen YE ; Yinfei ZHANG ; Chao LIU ; Youmin WANG ; Shengli WU ; Tao YANG ; Huacong DENG ; Lulu CHEN ; Tianshu ZENG ; Jiajun ZHAO ; Yiming MU ; Weiqing WANG ; Guang NING ; Jieli LU ; Min XU ; Yufang BI ; Weiguo HU
Frontiers of Medicine 2025;19(1):79-89
This study aimed to comprehensively examine the association of gallstones, cholecystectomy, and cancer risk. Multivariable logistic regressions were performed to estimate the observational associations of gallstones and cholecystectomy with cancer risk, using data from a nationwide cohort involving 239 799 participants. General and gender-specific two-sample Mendelian randomization (MR) analysis was further conducted to assess the causalities of the observed associations. Observationally, a history of gallstones without cholecystectomy was associated with a high risk of stomach cancer (adjusted odds ratio (aOR)=2.54, 95% confidence interval (CI) 1.50-4.28), liver and bile duct cancer (aOR=2.46, 95% CI 1.17-5.16), kidney cancer (aOR=2.04, 95% CI 1.05-3.94), and bladder cancer (aOR=2.23, 95% CI 1.01-5.13) in the general population, as well as cervical cancer (aOR=1.69, 95% CI 1.12-2.56) in women. Moreover, cholecystectomy was associated with high odds of stomach cancer (aOR=2.41, 95% CI 1.29-4.49), colorectal cancer (aOR=1.83, 95% CI 1.18-2.85), and cancer of liver and bile duct (aOR=2.58, 95% CI 1.11-6.02). MR analysis only supported the causal effect of gallstones on stomach, liver and bile duct, kidney, and bladder cancer. This study added evidence to the causal effect of gallstones on stomach, liver and bile duct, kidney, and bladder cancer, highlighting the importance of cancer screening in individuals with gallstones.
Humans
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Mendelian Randomization Analysis
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Gallstones/complications*
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Female
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Male
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Cholecystectomy/statistics & numerical data*
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Middle Aged
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Risk Factors
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Aged
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Adult
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Neoplasms/etiology*
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Stomach Neoplasms/epidemiology*
3.Guideline for Adult Weight Management in China
Weiqing WANG ; Qin WAN ; Jianhua MA ; Guang WANG ; Yufan WANG ; Guixia WANG ; Yongquan SHI ; Tingjun YE ; Xiaoguang SHI ; Jian KUANG ; Bo FENG ; Xiuyan FENG ; Guang NING ; Yiming MU ; Hongyu KUANG ; Xiaoping XING ; Chunli PIAO ; Xingbo CHENG ; Zhifeng CHENG ; Yufang BI ; Yan BI ; Wenshan LYU ; Dalong ZHU ; Cuiyan ZHU ; Wei ZHU ; Fei HUA ; Fei XIANG ; Shuang YAN ; Zilin SUN ; Yadong SUN ; Liqin SUN ; Luying SUN ; Li YAN ; Yanbing LI ; Hong LI ; Shu LI ; Ling LI ; Yiming LI ; Chenzhong LI ; Hua YANG ; Jinkui YANG ; Ling YANG ; Ying YANG ; Tao YANG ; Xiao YANG ; Xinhua XIAO ; Dan WU ; Jinsong KUANG ; Lanjie HE ; Wei GU ; Jie SHEN ; Yongfeng SONG ; Qiao ZHANG ; Hong ZHANG ; Yuwei ZHANG ; Junqing ZHANG ; Xianfeng ZHANG ; Miao ZHANG ; Yifei ZHANG ; Yingli LU ; Hong CHEN ; Li CHEN ; Bing CHEN ; Shihong CHEN ; Guiyan CHEN ; Haibing CHEN ; Lei CHEN ; Yanyan CHEN ; Genben CHEN ; Yikun ZHOU ; Xianghai ZHOU ; Qiang ZHOU ; Jiaqiang ZHOU ; Hongting ZHENG ; Zhongyan SHAN ; Jiajun ZHAO ; Dong ZHAO ; Ji HU ; Jiang HU ; Xinguo HOU ; Bimin SHI ; Tianpei HONG ; Mingxia YUAN ; Weibo XIA ; Xuejiang GU ; Yong XU ; Shuguang PANG ; Tianshu GAO ; Zuhua GAO ; Xiaohui GUO ; Hongyi CAO ; Mingfeng CAO ; Xiaopei CAO ; Jing MA ; Bin LU ; Zhen LIANG ; Jun LIANG ; Min LONG ; Yongde PENG ; Jin LU ; Hongyun LU ; Yan LU ; Chunping ZENG ; Binhong WEN ; Xueyong LOU ; Qingbo GUAN ; Lin LIAO ; Xin LIAO ; Ping XIONG ; Yaoming XUE
Chinese Journal of Endocrinology and Metabolism 2025;41(11):891-907
Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.
4.Deep learning-based segmentation method of neck skeletal muscle in radiotherapy patients with head and neck tumors
Zhi MING ; Ke LIU ; Bin ZENG ; Zhe WU ; Mu-jun LIU
Chinese Medical Equipment Journal 2025;46(8):11-17
Objective To propose a lightweight deep learning network-based segmentation method for automatic segmenta-tion of the skeletal muscle at the third cervical spine(C3)level.Methods Firstly,121 patients with head and neck tumors admitted to the Department of Oncology of Zigong First People's Hospital from January 2019 to December 2022 were selected and randomly divided into a training set,a validation set and a test set in the ratio of 7∶1∶2.Secondly,a lightweight Mamba architecture was introduced into the UNet network and an attention gate(AG)mechanism was added to the skip connection path to construct a MB-UNet network model.Finally,the trained network models were evaluated for segmentation performance on the test set,the MB-UNet network model was compared with manual segmentation over the results of determination of skeletal muscle area(SMA),and with classical network models in terms of parameter scale and computation effort including UNet,Deeplab V3+,U2Net,VMUNet and UltraLight-VMUNet models.The time required by the MB-UNet network model for predicting SMA and that by the physician with the assistance of the model was summarized.Results When used for segmenting the skeletal muscle at C3 level the constructued MB-UNet network model gained advantages over the classical models,with a Dice similarity coefficient of 88.23%,an intersection over union(IoU)ratio of 78.94%,a sensitivity of 91.27%and a 95%Hausdorff distance of 7.13 mm;the SMA determined by manual segementation was basically close to that by the MB-UNet network model;the MB-UNet network model behaved generally better than the classical network models,with the computation effort being 1.88 GFLOPS and the parameter scale being 0.77M;it took the MB-UNet network model 1.93 s for the prediction on the test set,and only 2 min for the physician to obtain satisfactory results with the assistance of the MB-UNet network model,which was significantly shorter than that by munual segmentation(20 min).Conclusion The proposed method contributes to segmenting the skeletal muscle at C3 level precisely and rapidly and calculating SMA accurately,which helps clinicians to quickly diagnose sarcopenia in patients with head and neck tumors and improves the diagnostic efficiency.[Chinese Medical Equipment Journal,2025,46(8):11-17]
5.Deep learning-based segmentation method of neck skeletal muscle in radiotherapy patients with head and neck tumors
Zhi MING ; Ke LIU ; Bin ZENG ; Zhe WU ; Mu-jun LIU
Chinese Medical Equipment Journal 2025;46(8):11-17
Objective To propose a lightweight deep learning network-based segmentation method for automatic segmenta-tion of the skeletal muscle at the third cervical spine(C3)level.Methods Firstly,121 patients with head and neck tumors admitted to the Department of Oncology of Zigong First People's Hospital from January 2019 to December 2022 were selected and randomly divided into a training set,a validation set and a test set in the ratio of 7∶1∶2.Secondly,a lightweight Mamba architecture was introduced into the UNet network and an attention gate(AG)mechanism was added to the skip connection path to construct a MB-UNet network model.Finally,the trained network models were evaluated for segmentation performance on the test set,the MB-UNet network model was compared with manual segmentation over the results of determination of skeletal muscle area(SMA),and with classical network models in terms of parameter scale and computation effort including UNet,Deeplab V3+,U2Net,VMUNet and UltraLight-VMUNet models.The time required by the MB-UNet network model for predicting SMA and that by the physician with the assistance of the model was summarized.Results When used for segmenting the skeletal muscle at C3 level the constructued MB-UNet network model gained advantages over the classical models,with a Dice similarity coefficient of 88.23%,an intersection over union(IoU)ratio of 78.94%,a sensitivity of 91.27%and a 95%Hausdorff distance of 7.13 mm;the SMA determined by manual segementation was basically close to that by the MB-UNet network model;the MB-UNet network model behaved generally better than the classical network models,with the computation effort being 1.88 GFLOPS and the parameter scale being 0.77M;it took the MB-UNet network model 1.93 s for the prediction on the test set,and only 2 min for the physician to obtain satisfactory results with the assistance of the MB-UNet network model,which was significantly shorter than that by munual segmentation(20 min).Conclusion The proposed method contributes to segmenting the skeletal muscle at C3 level precisely and rapidly and calculating SMA accurately,which helps clinicians to quickly diagnose sarcopenia in patients with head and neck tumors and improves the diagnostic efficiency.[Chinese Medical Equipment Journal,2025,46(8):11-17]
6.Comparison on imaging quality and semi-quantitative parameters of 18F-FDG whole-body images obtained with domestic NeuWise and Philips Ingenuity TF PET/CT scanners
Biyun MO ; Xingyu MU ; Jie QIN ; Yulong ZENG ; Weixia CHONG ; Nan LI ; Wei FU
Chinese Journal of Medical Imaging Technology 2025;41(5):816-820
Objective To compare imaging quality and semi-quantitative parameters of 18F-FDG whole-body images obtained with domestic NeuWise and Philips Ingenuity TF PET/CT scanners.Methods Thirty-four patients who underwent 18F-FDG whole-body scanning using NeuWise and Philips Ingenuity TF PET/CT systems respectively on the same day were enrolled.The imaging quality and semi-quantitative parameters of 2 kind images,also the mean standard uptake value(SUVmean)of normal tissue,the maximum standard uptake value(SUVmax),peak standard uptake value(SUVpeak),SUVmean of lesions,total lesion glycolysis(TLG)and metabolic tumor volume(MTV)were compared.Results No significant difference of imaging quality nor semi-quantitative parameters of lesions(all P>0.05),while significant differences of SUVmean of aortic arch,liver,lumbar vertebra and spinal cord were found between 2 kind images(all P<0.05).Strong correlations of SUVmax,SUVmean,MTV and TLG of lesions(r,=0.734-0.890,P<0.001),and high correlation of SUVpeak(rs=0.919,P<0.001)were found between 2 kind images.The consistency of SUVmax,SUVpeak,SUVmean,TLG and MTV at the lesion site between 2 kind images were very good to extremely good(ICC=0.891-0.986,all P<0.001),and the differences of all above semi-quantitative parameters were within 95%confidence interval.Conclusion Imaging quality of 18F-FDG whole-body images obtained with domestic NeuWise and Philips Ingenuity TF PET/CT scanners could meet the requirements of clinical diagnosis and treatment,and semi-quantitative parameters obtained based on both images had good consistencies.
7.Comparison on imaging quality and semi-quantitative parameters of 18F-FDG whole-body images obtained with domestic NeuWise and Philips Ingenuity TF PET/CT scanners
Biyun MO ; Xingyu MU ; Jie QIN ; Yulong ZENG ; Weixia CHONG ; Nan LI ; Wei FU
Chinese Journal of Medical Imaging Technology 2025;41(5):816-820
Objective To compare imaging quality and semi-quantitative parameters of 18F-FDG whole-body images obtained with domestic NeuWise and Philips Ingenuity TF PET/CT scanners.Methods Thirty-four patients who underwent 18F-FDG whole-body scanning using NeuWise and Philips Ingenuity TF PET/CT systems respectively on the same day were enrolled.The imaging quality and semi-quantitative parameters of 2 kind images,also the mean standard uptake value(SUVmean)of normal tissue,the maximum standard uptake value(SUVmax),peak standard uptake value(SUVpeak),SUVmean of lesions,total lesion glycolysis(TLG)and metabolic tumor volume(MTV)were compared.Results No significant difference of imaging quality nor semi-quantitative parameters of lesions(all P>0.05),while significant differences of SUVmean of aortic arch,liver,lumbar vertebra and spinal cord were found between 2 kind images(all P<0.05).Strong correlations of SUVmax,SUVmean,MTV and TLG of lesions(r,=0.734-0.890,P<0.001),and high correlation of SUVpeak(rs=0.919,P<0.001)were found between 2 kind images.The consistency of SUVmax,SUVpeak,SUVmean,TLG and MTV at the lesion site between 2 kind images were very good to extremely good(ICC=0.891-0.986,all P<0.001),and the differences of all above semi-quantitative parameters were within 95%confidence interval.Conclusion Imaging quality of 18F-FDG whole-body images obtained with domestic NeuWise and Philips Ingenuity TF PET/CT scanners could meet the requirements of clinical diagnosis and treatment,and semi-quantitative parameters obtained based on both images had good consistencies.
8.Guideline for Adult Weight Management in China
Weiqing WANG ; Qin WAN ; Jianhua MA ; Guang WANG ; Yufan WANG ; Guixia WANG ; Yongquan SHI ; Tingjun YE ; Xiaoguang SHI ; Jian KUANG ; Bo FENG ; Xiuyan FENG ; Guang NING ; Yiming MU ; Hongyu KUANG ; Xiaoping XING ; Chunli PIAO ; Xingbo CHENG ; Zhifeng CHENG ; Yufang BI ; Yan BI ; Wenshan LYU ; Dalong ZHU ; Cuiyan ZHU ; Wei ZHU ; Fei HUA ; Fei XIANG ; Shuang YAN ; Zilin SUN ; Yadong SUN ; Liqin SUN ; Luying SUN ; Li YAN ; Yanbing LI ; Hong LI ; Shu LI ; Ling LI ; Yiming LI ; Chenzhong LI ; Hua YANG ; Jinkui YANG ; Ling YANG ; Ying YANG ; Tao YANG ; Xiao YANG ; Xinhua XIAO ; Dan WU ; Jinsong KUANG ; Lanjie HE ; Wei GU ; Jie SHEN ; Yongfeng SONG ; Qiao ZHANG ; Hong ZHANG ; Yuwei ZHANG ; Junqing ZHANG ; Xianfeng ZHANG ; Miao ZHANG ; Yifei ZHANG ; Yingli LU ; Hong CHEN ; Li CHEN ; Bing CHEN ; Shihong CHEN ; Guiyan CHEN ; Haibing CHEN ; Lei CHEN ; Yanyan CHEN ; Genben CHEN ; Yikun ZHOU ; Xianghai ZHOU ; Qiang ZHOU ; Jiaqiang ZHOU ; Hongting ZHENG ; Zhongyan SHAN ; Jiajun ZHAO ; Dong ZHAO ; Ji HU ; Jiang HU ; Xinguo HOU ; Bimin SHI ; Tianpei HONG ; Mingxia YUAN ; Weibo XIA ; Xuejiang GU ; Yong XU ; Shuguang PANG ; Tianshu GAO ; Zuhua GAO ; Xiaohui GUO ; Hongyi CAO ; Mingfeng CAO ; Xiaopei CAO ; Jing MA ; Bin LU ; Zhen LIANG ; Jun LIANG ; Min LONG ; Yongde PENG ; Jin LU ; Hongyun LU ; Yan LU ; Chunping ZENG ; Binhong WEN ; Xueyong LOU ; Qingbo GUAN ; Lin LIAO ; Xin LIAO ; Ping XIONG ; Yaoming XUE
Chinese Journal of Endocrinology and Metabolism 2025;41(11):891-907
Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.
9.Shujin Jiannao Prescription alleviates inflammation in the cerebral cortex of rats with hypoxic-ischemic cerebral palsy
Gang LIU ; Jie ZENG ; Yalin ZHAO ; Bowen DENG ; Shengyuan JIANG ; Yaqi ZHANG ; Yi ZHAO ; Jingpei REN ; Chuanyu HU ; Lin XU ; Xiaohong MU
Chinese Journal of Tissue Engineering Research 2024;28(23):3674-3679
BACKGROUND:Shujin Jiannao Prescription is an empirical formula for the treatment of cerebral palsy in Dongzhimen Hospital,Beijing University of Chinese Medicine,with clear clinical efficacy,but the specific mechanism needs to be elucidated. OBJECTIVE:To explore the possible mechanism of Shujin Jiannao Prescription in treating cerebral palsy. METHODS:Sixty-four 7-day-old Sprague-Dawley rats were randomly divided into a normal group(n=12)and a model group(n=52).An animal model was established by the Rice-Vannucci method.After successful modeling,52 model rats were randomly divided into control model group(n=12),minocycline group,and the low-,medium-,and high-dose groups of the Shujin Jiannao Prescription(n=10 per group).Rats in the minocycline group were given 40 mg/kg·d minocycline by gavage;rats in the low-,medium,and high-dose groups were given 4,8,and 16 g/kg·d Shujin Jiannao Prescription granules by gavage,respectively;and rats in the normal group and control model group were given an equal dose of normal saline by gavage.Medication in each group was given once a day for 1 week.The rats in each group were evaluated behaviorally using suspension test,abnormal involuntary movement score,and Bederson score.The pathological changes of the cerebral cortex were observed by hematoxylin-eosin staining.The levels of tumor necrosis factor α,interleukin 1β,and interleukin 10 in the cerebral cortex were determined using ELISA.The positive expressions of Janus kinase 2(JAK2),phosphorylated Janus kinase 2(p-JAK2),phosphorylated signal transducer and activator of transcription 3(p-STAT3)in the cerebral cortex were detected using immunohistochemistry.The protein expression levels of JAK2,p-JAK2,and p-STAT3 were detected using western blot. RESULTS AND CONCLUSION:Compared with the normal group,the suspension test score and involuntary movement score were decreased in the control model group(P<0.01 or P<0.05).The pathological results showed structural disruption of nerve cells,formation of large numbers of vacuoles,cell swelling,and increased intercellular space in the control model group.In addition,the expressions of tumor necrosis factor α and interleukin 1β in the cerebral cortex were significantly increased(P<0.01),the expression of interleukin 10 was decreased(P<0.05),and the protein expressions of JAK2,p-JAK2,and p-STAT3 in the cerebral cortex were significantly increased(P<0.01)in the control model group compared with the normal group.Compared with the model group,minocycline and Shujin Jiannao Prescription at each dose could improve the behavioral indexes of rats(P<0.01 or P<0.05)and ischemic-hypoxic pathological changes were attenuated,with only a small amount of necrotic nerve cells and a few vacuoles,and reduced intercellular space.Moreover,the expressions of tumor necrosis factor α and interleukin 1β in the cerebral cortex were decreased in each drug group compared with the control model group(P<0.05),while the protein expressions of JAK2,p-JAK2,and p-STAT3 in the cerebral cortex were significantly decreased(P<0.01).The most obvious improvement was observed in the high-dose Shujin Jiannao Prescription group.To conclude,Shujin Jiannao Prescription can inhibit inflammation in the cerebral cortex of rats with hypoxic-ischemic brain injury.The mechanism may be related to the regulation of the JAK2/STAT3 signaling pathway.
10.Effects of Shujin Jiannao Prescription on cell apoptosis in rats with hypoxic-ischemic brain injury
Yu JIANG ; Lin XU ; Yalin ZHAO ; Gang LIU ; Yaqi ZHANG ; Huizhong BAI ; Jingpei REN ; Jie ZENG ; Xiaohong MU
Chinese Journal of Tissue Engineering Research 2024;28(28):4477-4483
BACKGROUND:Perinatal hypoxic-ischemic brain injury is one of the most common causes of cerebral palsy.Shujin Jiannao Prescription is an experienced formula for treating cerebral palsy and improving blood supply to the brain developed by the Dongzhimen Hospital,Beijing University of Chinese Medicine. OBJECTIVE:To explore the possible mechanism of Shujin Jiannao Prescription in treating hypoxic-ischemic cerebral palsy. METHODS:Sixty-four 7-day-old Sprague-Dawley rats were randomly divided into six groups.There were 12 rats in each of the control and model groups as well as 10 animals in each of the minocycline group,and the low-,medium-,and high-dose groups of Shujin Jiannao Prescription.The neonatal rat ischemic-hypoxic cerebral palsy model was established in all groups except for the control group.After successful modeling,rats in each drug group were respectively gavaged with minocycline and Shujin Jiannao Prescription at a dose of 4,8,and 16 g/kg per day for 1 week.Body mass of rats was measured and behavioral changes were detected before and after drug administration.Hematoxylin-eosin staining was used to observe the histomorphology of hippocampal CA1 region of rat brain tissue,and immunohistochemistry and western blot were used to detect the expression levels of Bcl-2,Bax,and Caspase-3 in the brain tissue of rats. RESULTS AND CONCLUSION:Compared with the model group,medium-and high-dose Shujin Jiannao Prescription significantly increased the body mass of rats(P<0.05).Compared with the model group,minocycline effectively prolonged the suspension time of ischemic-hypoxic cerebral palsy rats(P<0.05),while medium-and high-dose Shujin Jiannao Prescription significantly prolonged the suspension time,shortened the inclined plane test time,and increased the Longa score of rats(P<0.05).The pathological results showed that after drug intervention,only a small number of neuronal cells in the brain tissue of rats were necrotic,the cells were more neatly arranged,the cell structure was more complete,and only part of the cell nuclei became smaller.Compared with the model group,minocycline and medium-and high-dose Shujin Jiannao Prescription reduced the expression of Bax Caspase-3(P<0.05),medium-and high-dose Shujin Jiannao Prescription increased the expression of Bcl-2(P<0.05),and Bcl-2/Bax protein expression was increased in minocycline and three Shujin Jiannao Prescription groups(P<0.05).In addition,the protein expression was increased in a dose-dependent manner after intervention with Shujin Jiannao Prescription,and there was no significant difference between the minocycline and three Shujin Jiannao Prescription groups(P>0.05).To conclude,the mechanism by which Shujin Jiannao Prescription treats ischemic-hypoxic cerebral palsy in rats may be to enhance the expression of anti-apoptotic protein Bcl-2,inhibit the expression of pro-apoptotic protein Bax,and reduce the expression of Caspase-3,ultimately inhibiting the apoptosis of hippocampal neuronal cells in rats with cerebral palsy.Within a certain range,the higher dose of Shujin Jiannao Prescription indicates the better therapeutic effect,and the high-dose Shujin Jiannao Prescription is as effective as minocycline.

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