1.Protective effect of sericin on streptozotocin-induced INS-1 cell damage by regulating PI3K/Akt/NF-κB signaling pathway through Akt1 and its mechanism
Cheng CHEN ; Jingyao LI ; Wanxiang HU ; Donghui LIU ; Zhihong CHEN
Journal of Jilin University(Medicine Edition) 2025;51(3):590-598
Objective:To discuss the effect of sericin on the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/nuclear factor-κB(NF-κB)signaling pathway and apoptosis in the streptozotocin(STZ)-damaged INS-1 cells,and to clarify its mechanism.Methods:The INS-1 cells were cultured with complete medium containing 0,0.1,0.3,1.0,3.0,and 10.0 μmol·L-1 Akt1 inhibitor A-674563,10 mmol·L-1 STZ,and 600 mg·L-1 sericin,and divided into 0,0.1,0.3,1.0,3.0,and 10.0 μmol·L-1 A-674563 groups,and the control group(complete medium without drugs)was set up.Cell counting kit-8(CCK-8)method was used to detect the survival rates of the INS-1 cells,and the half-maximal inhibitory concentration(IC50)value was calculated to determine the optimal inhibitory concentration of A-674563,which was further verified by Western blotting method.The INS-1 cells were divided into normal control group(complete medium),model group(10 mmol·L-1 STZ+complete medium),and low,medium,and high doses of sericin groups(10 mmol·L-1 STZ+150 mg·L-1 sericin+complete medium,10 mmol·L-1 STZ+300 mg·L-1 sericin+complete medium,and 10 mmol·L-1 STZ+600 mg·L-1 sericin+complete medium).CCK-8 method was used to detect the survival rates of the INS-1 cells in various groups to determine the optimal concentration of sericin.Additionally,the INS-1 cells were divided into normal control group(complete medium),model group(10 mmol·L-1 STZ+complete medium),sericin group(10 mmol·L-1 STZ+600 mg·L-1 sericin+complete medium),and A-674563 group(10 mmol·L-1 STZ+600 mg·L-1 sericin+0.3 μmol·L-1 A-674563+complete medium).Flow cytometry was used to detect the apoptotic rates of the INS-1 cells in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of Akt1,NF-κB,tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)mRNA in the INS-1 cells in various groups;Western blotting method was used to detect the expression levels of phosphorylated Akt1(p-Akt1)and NF-κB proteins in the INS-1 cells in various groups;enzyme linked immunosorbent assay(ELISA)method was used to detect the levels of TNF-α and IL-6 in the INS-1 cells in various groups.Results:The survival rates of the INS-1 cells in control group was 100.00%±0.00%;in 0,0.1,0.3,1.0,3.0,and 10.0 μmol·L-1 A-674563+10 mmol·L-1 STZ+600 mg·L-1 sericin+complete medium groups,which were 82.50%±2.28%,69.47%±1.94%,51.51%±1.74%,38.94%±1.57%,24.79%±1.14%,and 19.85%±1.03%,respectively.The IC?? value of A-674563 for INS-1 cells was 0.3 μmol·L-1,and 0.3 μmol·L-1 A-674563 was selected for subsequent experiments.Compared with 0 μmol·L-1 A-674563,the expression level of p-Akt1 protein in the INS-1 cells after treated with 0.3 μmol·L-1 A-674563+10 mmol·L-1 STZ+600 mg·L-1 sericin+complete medium was significantly decreased(P<0.05).The CCK-8 results showed that compared with normal control group,the survival rate of the INS-1 cells in model group was significantly decreased(P<0.05);compared with model group,the survival rates of the INS-1 cells in low,medium,and high doses of sericin groups were significantly increased(P<0.05);compared with low and medium doses of sericin groups,the survival rate of the INS-1 cells in high dose of sericin group was significantly increased(P<0.05).Thus,600 mg·L-1 sericin was selected for subsequent experiments.The CCK-8 results showed that compared with normal control group,the survival rate of the INS-1 cells in model group was significantly decreased(P<0.05);compared with model group,the survival rate of the INS-1 cells in sericin group was significantly increased(P<0.05);compared with sericin group,the survival rate of the INS-1 cells in A-674563 group was significantly decreased(P<0.05).The flow cytometry results showed that compared with normal control group,the apoptotic rate of the INS-1 cells in model group was significantly increased(P<0.05);compared with model group,the apoptotic rate of the INS-1 cells in sericin group was significantly decreased(P<0.05);compared with sericin group,the apoptotic rate of the INS-1 cells in A-674563 group was significantly increased(P<0.05).The RT-qPCR results showed that compared with normal control group,the expression level of Akt1 mRNA in the INS-1 cells in model group was significantly decreased(P<0.05);compared with model group,the expression levels of Akt1 mRNA in the INS-1 cells in low,medium,and high doses of sericin groups were significantly increased(P<0.05);compared with low and medium doses of sericin groups,the expression level of Akt1 mRNA in the INS-1 cells in high dose of sericin group was significantly increased(P<0.05).Compared with normal control group,the expression levels of NF-κB,TNF-α,and IL-6 mRNA in the INS-1 cells in model group were significantly increased(P<0.05);compared with model group,the expression levels of NF-κB,TNF-α,and IL-6 mRNA in the INS-1 cells in sericin group were significantly decreased(P<0.05);compared with sericin group,the expression level of NF-κB mRNA in the INS-1 cells in A-674563 group was significantly increased(P<0.05).The Western blotting results showed that compared with normal control group,the expression level of p-Akt1 protein in the INS-1 cells in model group was significantly decreased(P<0.05);compared with model group,the expression levels of p-Akt1 protein in the INS-1 cells in low,medium,and high doses of sericin groups were significantly increased(P<0.05);compared with low and medium doses of sericin groups,the expression level of p-Akt1 protein in the INS-1 cells in high dose of sericin group was significantly increased(P<0.05).Compared with normal control group,the expression level of NF-κB protein in the INS-1 cells in model group was significantly increased(P<0.05);compared with model group,the expression level of NF-κB protein in the INS-1 cells in sericin group was significantly decreased(P<0.05);compared with sericin group,the expression level of NF-κB protein in the INS-1 cells in A-674563 group was significantly increased(P<0.05).The ELISA results showed that compared with normal control group,the levels of TNF-α and IL-6 in the INS-1 cells in model group were significantly increased(P<0.05);compared with model group,the levels of TNF-α and IL-6 in the INS-1 cells in sericin group were significantly decreased(P<0.05);compared with sericin group,the levels of TNF-α and IL-6 in the INS-1 cells in A-674563 group were significantly increased(P<0.05).Conclusion:Sericin alleviates the PI3K/Akt/NF-κB signaling pathway-mediated inflammatory response and apoptosis by targeting Akt1,exerting a protective effect against STZ-induced damage in INS-1 cells.
2.Improvement and development direction of bariatric metabolic surgery
Jingang LIU ; Jingyao HU ; Fu CHEN
Chinese Journal of Digestive Surgery 2023;22(1):94-99
In more than half a century of the development of bariatric metabolic surgery, a variety of classic surgical methods have been formulated. However, the improvement and innovation of bariatric metabolic surgery has never stopped. The replacement of new and old surgical methods in clinical application and development reflects the vitality and progress in the field of bariatric metabolic surgery, and also promotes the development of bariatric metabolic surgery to the best balance between benefits and risks. In the early stages, studies in metabolic surgery are more inclined to confirm the efficacy, safety and mechanism of classical procedures. In recent years, metabolic surgeons around the world have become more inclined to focus on the exploration and innovation of new procedures. In addition, the improvement of biliopancreatic diversion with duodenal switch and the sleeve gastrectomy plus procedures have gradually become hot spots for surgical innovation. However, the new techniques are diverse, scattered and partially overlapping. The authors make a comment on this content, in order to provide assistance to clinical and scientific research.
3.Necessities and paths to quality improvement of standardized construction of metabolic and bariatric surgery in China
Jingang LIU ; Jingyao HU ; Zhengyin WANG
Chinese Journal of Gastrointestinal Surgery 2023;26(11):1023-1027
Recently, the number of severe obesity in China has now ranked first in the world. The amount of metabolic and bariatric surgery in China is increasing year by year, and has made rapid development. As more and more new hospitals, surgical teams, and physicians join the field of metabolic and bariatric surgery, suboptimal operations and managements will inevitably accompany, causing problems and hidden dangers related to bariatric surgery. To a certain extent, this is in line with the law of development, but it does not mean that we can leave it alone and let it develop. In order to ensure the sustainable, healthy and orderly development of metabolic and bariatric surgery in China in the future, the standardized construction and quality improvement have become an urgent task. This paper reviews the current status of standardized construction of metabolic and bariatric surgery at home and abroad, the necessities and paths to quality improvement of standardized construction of metabolic and bariatric surgery in China, in order to put forward some thoughts and arouse extensive discussions for the development of the subject.
4.Necessities and paths to quality improvement of standardized construction of metabolic and bariatric surgery in China
Jingang LIU ; Jingyao HU ; Zhengyin WANG
Chinese Journal of Gastrointestinal Surgery 2023;26(11):1023-1027
Recently, the number of severe obesity in China has now ranked first in the world. The amount of metabolic and bariatric surgery in China is increasing year by year, and has made rapid development. As more and more new hospitals, surgical teams, and physicians join the field of metabolic and bariatric surgery, suboptimal operations and managements will inevitably accompany, causing problems and hidden dangers related to bariatric surgery. To a certain extent, this is in line with the law of development, but it does not mean that we can leave it alone and let it develop. In order to ensure the sustainable, healthy and orderly development of metabolic and bariatric surgery in China in the future, the standardized construction and quality improvement have become an urgent task. This paper reviews the current status of standardized construction of metabolic and bariatric surgery at home and abroad, the necessities and paths to quality improvement of standardized construction of metabolic and bariatric surgery in China, in order to put forward some thoughts and arouse extensive discussions for the development of the subject.
5.Clinical difference analysis and solution of lipid target and goal cut-off point determination of blood lipid management from different detection systems
Ruohong CHEN ; Fengxi WU ; Jingyao CAI ; Yiru ZHANG ; Zhifang ZHOU ; Min HU
Chinese Journal of Laboratory Medicine 2023;46(7):689-696
Objective:The results of the three lipid detection systems were compared to analyze their influence on risk stratification and clinical treatment in lipid management, especially the target goal cut-off point determination, and to find ways to reduce the impact on target goal determination of various lipid measurement system.Methods:A total of 196 serum samples with triglyceride TG <4.5 mmol/L were collected from people undergoing physical examinations and in-patients in the Second Xiangya Hospital of Central South University from August to October 2022. Triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were directly detected with Hitachi-Woke (HW), Roche and Mindray detection systems, respectively. The non high-density lipoprotein cholesterol (non HDL-C) was calculated by formula (TC-HDL-C) and LDL-C (F-LDL-C) was calculated by Friedewald formula, and results from various methodology were compared. The coefficient of variation ( CV) of these six indicators derived from the three detection systems were calculated to evaluate the consistency of the obtained results from different venders. In addition, the Pearson correlation coefficient was analyzed to evaluate the correlation of each indicator among different systems. According to the Chinese Guidelines for Blood Lipid Management, samples were divided into groups with LDL-C levels of <1.4, 1.4-<1.8, 1.8-<2.6, 2.6-<3.4 and ≥3.4 mmol/L according to the recommended LDL-C levels for different risk stratification levels. The sample size and percentage of LDL-C test results from different systems in the same group were counted to evaluate the impact of LDL-C differences between systems on clinical decision-making of blood lipid management. The correction factor was calculated through two methods: (1) The average deviation of LDL-C between systems was estimated by EP9-A3 method; (2) Multiple linear stepwise regression was used to establish the regression model of LDL-C difference and related indexes between systems. The two correction factors were used to correct the deviation of LDL-C value obtained from various systems, and Chi-square test was used to compare the difference of LDL-C grouping consistency rate before and after correction. Result:The average CV values of TG, TC, LDL-C, F-LDL-C, HDL-C, and non HDL-C among the three detection systems were 4.84%, 1.92%, 11.96%, 3.81%, 5.82% and 2.61%, respectively. Correlation analysis showed that when comparing the three systems in pairs, except for LDL-C derived from HW and Roche′s, and Mindray and Roche′s LDL-C ( R 2=0.938 and 0.947), the R 2 of other indicators were all greater than 0.97. The consistency rates of the three systems on LDL-C and F-LDL-C were 51.0% (100/196) and 90.8% (178/196), respectively, according to the risk stratification standard values and the difference was statistically significant ( P<0.05). When comparing in pairs, the consistency rates of Roche and HW, Mindray and HW, Mindray and Roche system LDL-C grouping were 60.7% (119/196), 82.7% (162/196), and 54.1% (106/196), respectively. After adjusting for mean deviation, the group consistency rate of Roche and HW increased to 73.7%-79.4% ( P<0.05), and the group consistency rate of Roche and Mindray increased to 72.3%-79.0% ( P<0.05). After adjusting for difference regression model, the group consistency rate of Roche and HW increased to 82.5%-84.0%, and the group consistency rate of Roche and Mindray increased to 81.0%-89.2%. However, there was no significant change in the group consistency rate of Mindray and HW after adjusting for both correction methods ( P>0.05) .Conclusions:There are significant differences in LDL-C derived from different detection systems, and the consistency rate of grouping according to the lipid-lowering standard value is relatively low, which may affect clinical decision-making in lipid management. Adjusted by the correction factor, the consistency rate of grouping between Roche and HW, Roche and Mindray systems with large differences in LDL-C can be improved. Using the difference multiple linear regression model as a correction factor is superior to the average deviation.
6.Exploration of cut-off values of amino acid levels in premature infants in Sichuan
Jingyao ZHOU ; Mingcai OU ; Xiaoju LUO ; Xingyue SU ; Yu ZHANG ; Qi HU ; Xuelian CHEN ; Lijuan YANG ; Yunxia YANG
Chinese Journal of Applied Clinical Pediatrics 2022;37(5):362-365
Objective:To detective the cut-off values of amino acid levels in premature infants in Sichuan.Methods:Data of newborns screening for inherited metabolic diseases (IMD) by tandem mass spectrometry in Sichuan Province from January 2018 to December 2019 were retrospectively analyzed.They were divided into premature infant group ( n=2 264, 1 312 males and 952 females) and full-term infant group ( n=53 275, 28 269 males and 25 006 females). The cut-off values of amino acids in dry blood spots were expressed as percentage ( P0.5 - P99.5), and rank sum test was used for comparison between preterm and full-term infants. Results:(1) The distribution of 11 amino acids [alanine (ALA), arginine (ARG), citrulline (CIT), glycine(GLY), leucine (LEU), methionine (MET), ornithine (ORN), phenylalanine (PHE), proline (PRO), tyrosine (TYR) and valine (VAL)] in premature infants were abnormal.(2) The cut-off values of amino acids in premature infants were as follows: ALA: 135.20-552.33 μmol/L, ARG: 1.34-47.04 μmol/L, CIT: 5.66-32.02 μmol/L, GLY: 181.48-909.93 μmol/L, LEU : 71.10-283.29 μmol/L, MET: 4.21-34.51 μmol/L, ORN: 40.58-293.76 μmol/L, PHE: 23.60-106.30 μmol/L, PRO: 77.76-358.24 μmol/L, TYR: 27.52-352.91 μmol/L, VAL: 53.74-228.37 μmol/L.(3) The cut-off values of amino acid in full-term infants were as follows: ALA: 135.20-552.33 μmol/L, ARG: 1.30-42.73 μmol/L, CIT: 5.92-30.35 μmol/L, GLY: 208.17-980.09 μmol/L, LEU: 72.91-287.49 μmol/L, MET: 4.27-33.90 μmol/L, ORN: 48.40-305.59 μmol/L, PHE: 27.63-92.27 μmol/L, PRO: 97.38-372.75 μmol/L, TYR: 40.19-276.54 μmol/L, VAL: 65.75-237.92 μmol/L.(4) Except for PHE ( Z=-0.58, P>0.05), the other indicators were significantly different between 2 groups [ALA ( Z=-15.32, P<0.05), ARG ( Z=-5.62, P<0.05), CIT ( Z=-5.86, P<0.05), GLY ( Z=-14.52, P<0.05), LEU ( Z=-5.62, P<0.05), MET ( Z=-5.22, P<0.05), ORN ( Z=-13.01, P<0.05), PRO ( Z=-22.09, P<0.05), TRY ( Z=-2.09, P<0.05), VAL ( Z=-17.82, P<0.05)]. Conclusions:The establishment of the cut-off values of amino acids in premature infants in Sichuan provides a theoretical basis for laboratory diagnosis of IMD screening, which enhances the accuracy of diagnosis and avoids excessive medical treatment.
7.The development of bariatric and metabolic surgery in China and the consideration of sur-gical complications
Jin'gang LIU ; Jingyao HU ; Yong ZHOU
Chinese Journal of Digestive Surgery 2022;21(1):57-61
Analysis of the layout and proportion of classic and innovative surgical procedures of bariatric and metabolic surgery throughout the world, the sleeve gastrectomy is beco-ming the most accepted and respected surgical procedure with its increasingly prominent advan-tages, and its operation number has even exceeded the "gold standard" surgery of the roux-en-Y gastric bypass. This trend is even more pronounced in China. With a small number of physicians lacking experience in gastrointestinal surgery and more and more young physicians joining in the field of bariatric and metabolic surgery, operation related complications and the consequences of insufficient perioperative management experience have occurred successively in clinical practice, such as gastrointestinal leakage, bleeding, lumen and anastomotic stenosis, internal hernia and obstruction, venous thromboembolism, etc. Bariatric and metabolic surgery is a branch of gastro-intestinal surgery, and physicians in the field should have the ability to prevent, predict and manage short- and long-term complications after surgery. The authors believe that through the comprehen-sive evaluation of laboratory indexes, patient signs and related examinations, physicians should have the basic qualities of decisively choosing the surgical timing, choosing the right decision-making, and taking effective measures.
8.Opportunities and challenges of bariatric and metabolic surgery in the future
Jingang LIU ; Yong ZHOU ; Jingyao HU
International Journal of Surgery 2021;48(5):289-294
Bariatric and metabolic surgery has been in China for more than 30 years, which has developed rapidly. However, it can be predicted that there will be many opportunities and challenges in the future development of bariatric and metabolic surgery, including the replacement of classic surgical methods, the guarantee of surgical safety, the brave exploration of new surgical methods, the decisive abandonment of bad surgical methods, the implementation of surgical standardization, and the breakthrough of basic and clinical research all mean more efforts. With the continuous rise of the incidence rate of metabolic syndrome and the continuous improvement of people′s health needs, these opportunities and challenges are particularly significant. Bariatric and metabolic surgery will also play a crucial role in overcoming obesity and metabolic diseases.
9.Clinical and genetic features of seven children with MYH9-related disease
Yu HU ; Jingyao MA ; Huiqing LIU ; Bixi YANG ; Shasha ZHAO ; Jialu ZHANG ; Runhui WU
Chinese Journal of Pediatrics 2021;59(11):968-972
Objective:To summarize and analyze of the clinical and genetic characteristics of children with nonmuscle myosin heavy chain 9 (MYH9)-related disease (MYH9-RD).Methods:To screen the patients who were first diagnosed as "chronic/refractory immune thrombocytopenia (ITP) " from April 2016 to May 2019 in Beijing Children′s Hospital by genetic and clinical examinations, then the clinical manifestation, laboratory examination and genetics results of 7 children diagnosed with MYH9-RD were collected and summarized retrospectively.Results:Among 7 children diagnosed with MYH9-RD, 3 were males and 4 females. The age of onset was 1.25 (0.41-6.16) years. The course of disease was 2.16 (0.41-8.59) years. The automatic platelet count was (9 (5-30))×10 9/L. All the cases were found with giant platelets under microscope,and the manual platelet count was (70 (30-100))×10 9/L. Four cases had skin hemorrhage or epistaxis and 3 cases had no bleeding. All 7 patients had received first-or second-line therapy of ITP, of whom 1 case received splenic embolization, and all the treatments mentioned above were ineffective. Finally, it was confirmed that all 7 patients had heterozygous missense mutations of MYH9 gene by next generation sequencing (NGS), including 2 pedigrees and 5 sporadic cases. Four sporadic mutations occurred in N-terminal globular head domain (HD), and 1 sporadic case with p.D1424N mutations occurred in the C-terminal tail domain (TD). One of the pedigrees also had p.D1424N mutation. The other familial case had a novel variant with one missense variant p.A44D caused by the c.131C>A transition. One of the two p.R702 mutations had kidney damage, and several relatives of the new p.A44D mutations had deafness. Conclusions:In this study, the spontaneous mutations of seven MYH9-RD were common, and all patients were misdiagnosed as ITP, whereas the bleeding was mild and immunotherapy was ineffective. The suspected disease can be identified earlier by manual visual platelet volume and count, which can be confirmed by genetic testing. It is more important to monitor the development of other organs damage instead of thrombocytopenia. For cases with p.R702 mutations the doctor should be aware of kidney damage, and for the cases with novel mutations p.A44D the doctor should be aware of hearing loss.
10.The influence of new surgical methods on the development of bariatric and metabolic surgery
Chinese Journal of Digestive Surgery 2020;19(11):1135-1139
Bariatric and metabolic surgery originated in the 1950s. After continuous verifications and improvements by bariatric and metabolic surgeons all over the world, bariatric surgery has been recognized as the 'gold standard’ for the treatment of metabolic diseases such as obesity. In the development of more than half a century, bariatric and metabolic surgery have formed a variety of classic surgical methods. At the same time, the new methods of bariatric surgery are emerging. Among them, the additional surgery based on sleeve gastrectomy has become a research hotspot. In the process of clinical applications of various new bariatric surgery methods, bariatric and metabolic surgeons should be objective, accurate, realistic, and move forward on the basis of criticizing and absorbing. The authors investigate the influence of new surgical methods on the development of bariatric and metabolic surgery.

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