1.Applicability of BlueScreen HC in genotoxicity detection of migrants mixtures of food contact materials
Min LI ; Jun LIN ; Weiliang WU ; Haixia SUI ; Xingfen YANG
Chinese Journal of Pharmacology and Toxicology 2024;38(10):796-806
OBJECTIVE To explore the applicablity of 'BlueScreen HC'(BSHC),a high throughput genotoxicity screening system based on human growth arrest and DNA damage inducible 45α(GADD45α)gene,in detecting the genotoxicity of migrants mixtures from food contact materials(FCM).METHODS The 2000 bp sequence upstream of the open reading frame of human GADD45α gene was used as the promoter to construct the lentiviral plasmid pEZX-LvPG04,which was double labeled by purinamycin and Gausluciferase(Gluc),and the lentiviral plasmid was infected with human lymphoblastocyte TK6 to obtain a stable transmutation cell line TK6-Gluc.Methyl methylate(MMS)at concentrations of 0,1.56,3.13,6.25,12.5,25.0 and 50.0 mg·L-1 was selected as the genotoxin without liver S9,cyclophosphamide(CTX)0,0.78,1.56,3.13,6.25,12.5,25.0 mg·L-1 was selected as the pre-genotoxin with liver S9,and dimethyl sulfoxide(DMSO)0,0.35,0.69,1.38,2.75,5.5 and 11.0 g·L-1 was selected as the non-genotoxin.The constructed BSHC was verified with the above known genetic positive and negative substance respectively.Polybutyleneadipate-co-terephthalate(MS/PBAT)was tested using 4% (V/V)acetic acid,and 10%,20%,50% and 95% (V/V)ethanol as food simulants at 40℃for 24 hours to obtain 5 multi-component migrants of MS/PBAT that were obtained by using DMSO as a solvent.TK6-Gluc cells were treated with 5 multi-component migrants of MS/PBAT at concentrations of 0,0.38,0.76,1.53,3.05,6.10 and 12.20 g·L-1 with or without liver S9.Cells were treated without liver S9 for 48 h.Cells treated with liver S9-mix were incubated for 3 h at a final concen-tration of 1% (V/V)liver S9 before being washed and re-suspended in fresh recovery media for another 45 h.After exposure,the cell viability was detected using the CCK-8 cell activity kit,and the Gluc Lumi-nescence in the medium was detected with Secrete-PairTM Gaussia Luciferase Assay Kit.In addition,the mutagenicity on Salmonella typhimurium TA98 and TA100 was detected by micro-fluctuation Ames test with 5 multi-component migrants of MS/PBAT at concentrations of 3.05 and 12.20 g·L-1.The in vitro mammalian cell chromosome aberration test was performed on CHL cells with 5 multi-component migrants of MS/PBAT at concentrations of 3.05 and 12.20 g·L-1 to detect the chromosomal aberration.The results of genotoxicity were compared with those of BSHC.RESULTS The lowest effect centra-tion(LEC;<80% relative cell viability)and the coytotoxicity(<30% relative cell viability)was defined.A positive genotoxicity result threshold was determined at 1.8-fold relative induction.For the liver S9 protocol,the same process was followed,and the decision threshold derived was 1.5-fold relative Gluc induction.It is considered as genetic substance only when a positive genotoxicity result was reached and there was no cytotoxicity.Compared with the vehicle control group,no genotoxicity was observed at all concentration of DMSO by BSHC.MMS 12.5,25.0 and 50.0 mg·L-1 produced genotoxicity without liver S9 while CTX 6.25,12.5 and 25.0 mg·L-1 produced genotoxicity with liver S9.Significant cell growth inhibition was observed in 95% ethanol migrants of MS/PBAT at concentrations of 6.10 and 12.20 g·L-1,and in 50% ethanol migrants of MS/PBAT at a concentration of 12.20 g·L-1 without liver S9.No cytotoxicity with a relative cell viability below 30% was observed in any of the treatment groups,and no high expression of Gluc was observed.Therefore,none of the 5 multi-component migrants produced genotoxicity without liver S9.Significant cell growth inhibition was observed in 95% ethanol migrants of MS/PBAT at a concentration of 12.20 g·L-1,and in 4% acetic acid migrants of MS/PBAT at concentrations of 6.10 and 12.20 g·L-1 with liver S9.No cytotoxicity with a relative cell viability below 30% was observed in any of the treatment groups.No high expression of Gluc was observed.There-fore,none of the 5 multi-component migrants produced genotoxicity with liver S9.In the micro fluctua-tion Ames test,when 5 multi-component migrants of MS/PBAT were treated with concentrations of 3.05 and 12.20 g·L-1 on TA98 and TA100 strains,there was no significant difference in the number of muta-genic positive wells compared with DMSO control group with or without liver S9,indicating that no mutagenic effect was produced.When CHL cells were treated with 5 multi-component migrants of MS/PBAT at concentration of 3.05 and 12.20 g·L-1,compared with DMSO control group,there was no signifi-cant difference in chromosome aberration rate of CHL cells with or without liver S9.CONCLUSION BSHC based on GADD45α gene has been established,which can be used for in vitro genotoxicity eval-uation of migrants mixtures of FCM,but further exploration of its minimum effective concentrations is still needed,and more types of mixtures need to be applied for further validation.
2.Development of Cognitive Assessment Scale for Spinal Cord Injury Rehabilitation and its reliability and validity
Haixia XIE ; Hua ZHAI ; Xinyu WANG ; Jun'an ZHOU ; Feng SHEN ; Airong WU ; Ying LIU ; Rundi CHEN ; Xuheng ZENG ; Peipei LIN ; Fengshui CHANG
Chinese Journal of Modern Nursing 2024;30(26):3507-3513
Objective:To develop a Cognitive Assessment Scale for Spinal Cord Injury (SCI) Rehabilitation and conduct reliability and validity tests in community-dwelling patients with SCI.Methods:Based on expectation value theory, social cognition theory, and goal setting theory, a Cognitive Assessment Scale for SCI Rehabilitation was developed through literature review, group discussions, patient trials, and expert verification. From February to December 2021, convenience sampling was used to select 231 community-dwelling patients with SCI as research subjects, including 67 community-dwelling patients with SCI who participated in rehabilitation training at Shanghai Sunshine Rehabilitation Center and 164 patients with SCI in the "Hope Home" WeChat group of Shanghai Sunshine Rehabilitation Center. Research subjects were surveyed using the Cognitive Assessment Scale for SCI Rehabilitation (patient version), 9-item depression scale of Patient Health Questionnaire, 7-item Generalized Anxiety Disorder Scale, EuroQol 5 Dimension-Visual Analogue Scale (EQ-VAS), General Self-Efficacy Scale, and general information questionnaire. SPSS 16.0 software and Amos 21.0 software were used for correlation analysis and reliability and validity testing.Results:The Cognitive Assessment Scale for SCI Rehabilitation (patient version) included two primary dimensions, eight secondary dimensions, and 24 items. The trial showed good results among patients with SCI and their caregivers, and experts generally agreed. Exploratory factor analysis found that the scale were divided into recognition dimension and understanding dimension. Cronbach's α coefficient of the scale was 0.98, the correlation coefficient between each item and its corresponding dimension was 0.75 to 0.88, and our results indicated good test-retest reliability. Correlation analysis showed that patient anxiety and depression scores were negatively correlated with rehabilitation cognitive scores ( P<0.05), and self-efficacy, quality of life were positively correlated with rehabilitation cognitive scores ( P<0.05) . Conclusions:The Cognitive Assessment Scale for SCI Rehabilitation is scientific and feasible, with good reliability and validity, and can be used to evaluate the rehabilitation cognition of community-dwelling patients with SCI.
3.Management of adult Langerhans cell histiocytosis in the hypothalamic-pituitary region—Experiences from Huashan Hospital
Qian WANG ; Quanya SUN ; Min HE ; Li PAN ; Yongfei WANG ; Haixia CHENG ; Yue WU ; Tianling DING ; Hongying YE ; Wei WU
Chinese Journal of Endocrinology and Metabolism 2024;40(5):386-392
Objective:To summarize experience of managing adult Langerhans cell histiocytosis(LCH) in hypothalamic-pituitary region(HPR) from Shanghai Huashan Hospital.Methods:Adult HPR-LCH patients diagnosed at oar endocrinology department from January 2013 to February 2022 were included. Clinical characteristics and treatment response were retrospectively analyzed.Results:A total of 27 adult HPR-LCH patients were included, with 14 cases involving the hypothalamus(H group) and 13 cases without(group NH). The common radiological findings included thickening of the pituitary stalk(25/27, 92.6%). At the time of diagnosis, 14 cases(51.9%) presented with panhypopituitarism, and 19 cases(70.4%) exhibited metabolic abnormalities. The group H had higher proportions of adrenal insufficiency, central hypothyroidism, panhypopituitarism, and diabetes compared to group NH(78.6% vs 23.1%; 78.6% vs 23.1%; 92.9% vs 30.8%, 35.7% vs 0%, respectively, all P<0.05). Hypothalamus syndrome was identified in 71.4%(10/14) of group H. The inital diagnosis rate was 79.2%(19/24), with 48.1% and 51.9% through biopsy of sellar and extrasellar lesions, respectively. Repeated biopsies confirmed the diagnosis in 25.9%(7/27) of cases. The peripheral lesions included bone, thyroid, lung, lymph node, thymus and liver. Out of 20 cases treated with chemotherapy, the objective response rate was 85% at 12 weeks. Four cases received local therapy, one case received traditional Chinese medicine treatment, one case abandoned treatment, and one case was lost to follow-up. The median follow-up time was 28(range 15 to 54) months. During this period, there were 3 deaths in group H and 1 death in group NH. Conclusion:Adult HPR-LCH patients presented with diabetes insipidus and high prevalences of hypopituitarism, hypothalamus syndrome and metabolic abnormalities. Typical imaging features were pituitary stalk thickening. A solitary mass in the HPR was usually very small, posing a great challenge for early diagnosis. Systemic evaluation would help to clarify the diagnosis. Patients with hypothalamus involvement had a higher mortality rate, suggesting the hypothalamus as a risk organ with poor prognosis.
4.Construction and application of an intelligent insurance medication audit system based on pre-auditor system
Xiaoyan WU ; Haixia ZHANG ; Mian ZHAO ; Xiaojie BIAN ; Jie PENG ; Hao ZHOU ; Bin ZHANG ; Weihong GE
Chinese Journal of Hospital Administration 2024;40(7):541-546
The establishment of an intelligent audit system for hospital medical insurance is crucial for standardizing irrational practices and curbing the unwarranted escalation of medical insurance costs. A tertiary hospital has developed comprehensive prescription review rules in terms of indications, nutritional risk assessments, pain score evaluations, drug sensitivity test results, and target patient populations. When using medical insurance funds for settlement, both systematic and manual reviews are conducted, and three-level intervention measures such as " prompt, review, and interception" are implemented based on prescription issues. After the system was put into use, the hospital′s in-process information system supervision mode shifted from the previous focus on prompts to focus on audits. The medical insurance intelligent audit system provided timely prompts and interventions, and the number of unreasonable medical insurance prescriptions and deductions from medical insurance sampling prescriptions significantly decreased ( P<0.001), while the number of medical insurance medication prescription audits increased ( P<0.001). The application of the intelligent medical insurance medication audit system has improved the efficiency of prescription auditing and ensured the rationality of the use of medical insurance funds.
5.Analysis of the therapeutic effect of avatinib bridged allogeneic hematopoietic stem cell transplantation on 7 cases of recurrent/refractory RUNX1-RUNX1T1 positive acute myeloid leukemia with KIT mutations
Siyi HAN ; Haixia ZHOU ; Yue HAN ; Depei WU
Chinese Journal of Hematology 2024;45(8):767-771
Objective:To evaluate the efficacy of avatinib plus allogeneic hematopoietic stem cell transplantation (allo-HSCT) for the treatment of recurrent/refractory RUNX1-RUNX1T1 positive acute myeloid leukemia (AML) with KIT mutations.Method:A retrospective study was conducted on the clinical data of seven relapsed/refractory AML patients containing the RUNX1-RUNX1T1 fusion gene and KIT mutation who received afatinib plus allo-HSCT treatment at the First Affiliated Hospital of Soochow University from June 2019 to June 2023.Results:The seven AML patients included one male and six females with a median age of 37 (18-56) years. All seven patients had KIT mutations (five positive for D816V and two positive for D816Y) . There were two refractory patients and five relapsed patients (all of whom had bone marrow recurrence) . All patients had to complete at least one course of treatment with afatinib before transplantation. Four patients achieved complete remission (CR) after treatment with afatinib, six patients had negative KIT gene mutations, and one had a decreased KIT gene mutational burden. There were three cases of unrelated identical transplantation and four cases of haploidentical transplantation. All patients received the modified Bu/Cy pretreatment regimen. After transplantation, all patients were successfully implanted and a bone marrow examination showed CR and minimal residual disease turned negative. Five patients exhibited negative fusion genes. Two patients died from infection following transplantation.Conclusion:Afatinib plus allo-HSCT may be an effective and safe new treatment strategy for RUNX1-RUNX1T1 positive AML patients with KIT-D816 mutation.
6.Comparison of the early postoperative visual quality of oval versus round valve in FS-LASIK
Xion-Gzi LIANG ; Qingsong ZHANG ; Wanju YANG ; Si LENG ; Haixia WU ; Minghui ZHANG
The Journal of Practical Medicine 2024;40(17):2430-2434
Objective To investigate the difference in visual quality between oval and round corneal flaps in femtosecond excimer laser in situ keratomileusis(FS-LASIK).Methods In a prospective randomized controlled clinical study,patients who voluntarily underwent FS-LASIK surgery in Wuhan University Affiliated Aier Ophthalmology Hospital from June 2022 to February 2023 were randomly divided into oval valve group and round valve group,with 36 patients(72 eyes)in each group.During FS-LASIK,oval and round corneal flaps were made,and excimer laser surgery was performed in parallel.Observe intraoperative and postoperative complications.The unaided visual acuity(UCVA)and spherical equivalent(SE)at 1 day,1 week,1 month and 3 months after surgery were compared between the two groups at 3 months after surgery,and the modulation transfer function(MTF),Strel ratio(SR)and total higher-order aberration at 3 months after surgery were compared between the two groups.Results All patients had successfully completed the surgery without serious complications.There was no significant difference between the oval valve group and the round valve group in UCVA at 1 day,1 week after surgery,1 month after surgery,3 months after surgery.There was no significant difference between the two groups on the postoperative SE at 1 day,1 week after surgery,1 month after surgery,and 3 months after surgery.There was no significant difference in MTF,SR and total higher-order aberration between the two groups at 3 months after surgery.Conclusion Ideal visual acuity can be achieved with either oval or round corneal flap during FS-LASIK,and there is no significant difference in visual quality between the two in the early postoperativeperiod.
7.Research Progress of Surfactants in Nanoformulation
Jingyu ZHOU ; He ZHANG ; Yi CHENG ; Meng LI ; Zengming WANG ; Nan LIU ; Haixia WU ; Hui ZHANG ; Aiping ZHENG
Herald of Medicine 2024;43(11):1790-1798
Surfactant is a kind of substance that can significantly reduce the surface/interface tension.Surfactant is an important substance that affects the preparation technology,stability and safety of nanoformulation.By combing the structure classification of surfactants and their function mechanism in nanoformulation,combined with the research status at home and abroad,the application and research of surfactants in the listed nanoformulation were described,in order to provide reference for the research and development of new nanoformulation.
8.Mechanism of the effect of Xuebijing injection on neurological function and survival of rats after cardiac arrest/cardiopulmonary resuscitation
Deqing HUANG ; Yuguang GAO ; Yuankan ZHANG ; Zhenglin WANG ; Haixia DENG ; Xiabing HUANG ; Yan PANG ; Lin WU
China Pharmacy 2024;35(6):653-658
OBJECTIVE To explore the potential mechanism of the effect of Xuebijing injection (XBJ) on neurological function and survival of rats after cardiac arrest (CA)/cardiopulmonary resuscitation (CPR) based on the S-nitrosoglutathione reductase (GSNOR)/S-nitrosoglutathione (GSNO) pathway. METHODS The CA/CPR rat model was established by ventricular fibrillation. Using a sham operation group as control, high-throughput sequencing was employed to analyze and mine the differentially expressed genes (DEGs). Enzyme-linked immunosorbent assay was used to determine the contents of GSNOR and GSNO in the hippocampus; the active components of XBJ were screened and subjected to molecular docking analysis with GSNOR. The rats successfully modeled using the same method were divided into model group (n=30), inhibitor (GSNOR inhibitor) group (n=30), XBJ group (n=30) and XBJ+inhibitor group (n=30), and a sham operation group (n=30) was set up. Neurological function was evaluated and survival status was recorded at 3 hours, 24 hours and 3 days after the first 89) drug intervention. The contents of GSNOR and GSNO in the hippocampus of rats were determined in each group at the 0191) above time points, and the relationship of the contents of GSNOR and GSNO with modified neurologic severity scale (mNSS) score was analyzed. RESULTS GSNOR coding gene was differentially expressed between the model group and the sham operation group. Compared with the sham operation group, GSNOR content increased significantly in the hippocampus of rats in model group, while GSNO content decreased significantly (P<0.05). The active components of XBJ, such as 4- methylenemiltirone and salviolone, could be bound to GSNOR protein, with the binding energy lower than -6 kcal/mol, mainly connected by hydrogen bonds. Animal experiments revealed that mNSS score and GSNOR levels in the hippocampus of rats in the model group were significantly higher than those in the sham operation group (P<0.05), while GSNO levels and survival rate were significantly lower than those in the sham operation group (P<0.05). The above indexes of rats were improved significantly in administration groups, the mNSS score in the XBJ group was significantly lower than that in the inhibitor group, the content changes of GSNOR and GSNO in the inhibitor group were more obvious than those in the XBJ group, and the various indicators in the XBJ+inhibitor group were significantly better than the XBJ group and the inhibitor group (P<0.05). GSNOR content was positively correlated with the mNSS score, and GSNO content was negatively correlated with the mNSS score (P<0.05). CONCLUSIONS XBJ can improve the neurological function of rats and enhance their survival rates after CA/CPR, the mechanism of which may be associated with the down-regulation of GSNOR and the up-regulation of GSNO.
9.Vascular endothelial growth factor combined with basic fibroblast growth factor improves replicative senescence of bone marrow mesenchymal stem cells
Weili SHI ; Shanshan LIU ; Hongbo CHANG ; Haixia GAO ; Xinzhou WANG ; Nan QIN ; Hong WU
Chinese Journal of Tissue Engineering Research 2024;28(31):4958-4963
BACKGROUND:Mesenchymal stem cells are susceptible to senescence during in vitro expansion,which greatly hinders their application in vivo and in vitro.How to improve the replicative senescence of mesenchymal stem cells is an urgent problem to be solved in tissue engineering. OBJECTIVE:To determine whether vascular endothelial growth factor combined with basic fibroblast growth factor can improve the aging of bone marrow mesenchymal stem cells caused by replicative passage. METHODS:Rat bone marrow mesenchymal stem cells were extracted by whole bone marrow adhesion method.Passage 2 cells were selected as normal control group.Passage 7 and later algebraic cells were selected as aging model group.Vascular endothelial growth factor(50 μg/L),basic fibroblast growth factor(10 μg/L),and their combination were administered.Cell proliferation was detected by CCK-8 assay.Cell senescence was observed by β-galactosidase activity staining.Cytoskeleton size and colony formation ability were observed by phalloidine staining and Giemsa staining,respectively,and the levels of senescence-related genes P16,P21,and P53 were detected by qRT-PCR.Gene expression levels of P16,P21,and P53 were tested by qRT-PCR. RESULTS AND CONCLUSION:(1)Vascular endothelial growth factor combined with basic fibroblast growth factor could promote the proliferation of aged bone marrow mesenchymal stem cells,which began to enter the plateau stage on day 9,and the absorbance value of the combined intervention group was significantly higher than that of the model group on day 9(P<0.05).(2)The phenotypic markers of the cells in the combined intervention group did not change,and the cell morphology changed from broad to slender.(3)Compared with the model group,the positive rate of β-galactosidase was significantly decreased(P<0.01);the number of nuclei increased(P<0.001);the total area of cytoskeleton increased(P<0.01);colony formation ability was enhanced(P<0.05);expression level of P16 was decreased(P<0.01)in the combined intervention group.These results indicate that vascular endothelial growth factor combined with basic fibroblast growth factor can improve the senescence of bone marrow mesenchymal stem cells caused by replicative passage without changing the cell phenotype.
10.Biological Connotation of Pathogenesis of Colorectal Cancer Due to Damp-heat Accumulation Based on Immune Response-intestinal Microbial Axis
Yijuan WU ; Xinghong SUN ; Haixia GUO ; Xiang'an ZAHNG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(6):228-237
Colorectal cancer (CRC) is a malignant tumor of the intestinal tract with changes in bowel habits, blood in the stool, and pain as the main clinical manifestations. With the change in lifestyle and diet structure in recent years, the incidence of CRC has been increasing year by year. The pathogenesis of CRC is closely related to abnormal immune response and chronic inflammation, intestinal microbial dysbiosis, and the production of oncogenic metabolites. There is a two-way communication between the intestinal microbiota and the body's immunity, which not only plays a key role in maintaining the body's health but also has a close relationship with the development of diseases. An increasing number of studies have shown that abnormal immune responses accelerate the disease process by producing inflammatory factors, causing chronic inflammation in the body, disrupting the intestinal mucosal barrier, and increasing mucosal permeability, thus resulting in dysbiosis of the intestinal microbial ecology and a large number of pathogenic microorganisms and their metabolites. In addition, dysbiosis of intestinal microbes, by suppressing the normal immune response, leads to the disruption of multiple metabolic pathways in the body, affecting the internal and external stress response of the intestine, inducing inflammation, and thus producing disease. Therefore, the complex crosstalk mechanism between the immune response and intestinal microbial axis is closely related to the development of CRC. Based on traditional Chinese medicine theory and clinical research, it was found that dietary factors are an important causative factor in the development of CRC. The deficiency of positive energy is the root cause of the disease, and damp-heat accumulation is the key pathogenesis. Through modern medical and biological research, it is believed that abnormal immune response is the microscopic manifestation of damp-heat entrapment, while intestinal microbial dysbiosis is the biological basis of toxic injection into the large intestine, and in the pathogenesis of CRC, the imbalance of immune response-intestinal microbial axis is compatible with damp-heat accumulation in traditional Chinese medicine. This study aims to explore the biological connotation of CRC due to damp-heat accumulation from the immune response-intestinal microbial axis, so as to interpret the pathogenesis of CRC due to damp-heat accumulation with objective data and provide new ideas and theoretical basis for the pathogenesis and treatment strategies of CRC due to damp-heat accumulation.

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