1.Predictive value of monocyte to lymphocyte ratio,neutrophil percentage to albumin ratio for diabetic macular edema
Shuning ZHAO ; Xiangling LIU ; Shaobo SU ; Xiaoqing WU
Journal of Xinxiang Medical College 2024;41(1):21-25,31
Objective To investigate the value of monocyte to lymphocyte ratio(MLR)and neutrophil percentage to albumin ratio(NPAR)in predicting diabetic macular edema(DME).Methods One hundred and one diabetic retinopathy patients admitted to the Third Affiliated Hospital of Xinxiang Medical University from January 2018 to February 2023 were selected as the research subjects,and they were divided into DME group(n=56)and non-DME group(n=45)based on fun-dus examination results.The general data such as gender,age,course of diabetes and laboratory indicators were collected by consulting medical records.Fasting elbow venous blood was collected early in the morning of the next day after the diagnosis of DME in both groups,the monocytes(MONO)count,lymphocyte(LYM)count,white blood cell(WBC)count,percentage of neutrophils(NEUT),plasma albumin(ALB),glycosylated haemoglobin(HbA1c)were measured by full automatic blood routine analyzer,and MLR,NPAR were calculated.General information and laboratory indexes of patients in the two groups were compared,and risk factors for DME were analyzed by multivariate logistic regression,and receiver operator characteristic(ROC)curve was applied to evaluate the predictive value of MLR and NPAR for DME.Results The course of diabetes,MONO count,NEUT,MLR,NPAR,WBC count,and HbA1c level of patients between the DME group were significantly higher than those in the non-DME group(P<0.05);there was no statistically significant difference in gender,age,LYM count,and ALB level of patients between the two groups(P>0.05).Multivariate logistic regression analysis showed that increased levels of WBC,MLR,and NPAR were independent risk factors for the occurrence of DME(P<0.05).The ROC curve showed that the best cut-off value of MLR was 0.192,and the area under the curve(AUC)for the prediction of DME was 0.729(95%confidence interval:0.631-0.826),with a sensitivity of 58.9%and a specificity of 82.2%;while the best cut-off value of NPAR was 1.404,and the AUC for predicting DME occurrence was 0.884(95%confidence interval:0.820-0.949),with a sensitivity of 75.0%and a specificity of 91.1%;the AUC of MLR and NPAP for predicting the occurrence of DME was 0.906(95%confidence interval:0.851-0.906),with a sensitivity of 69.6%and a specificity of 93.3%.With MLR>0.192 as positive and NPAR>1.404 as positive,the parallel test of MLR and NPAR predicted the occurrence of DME with a sensitivity of 87.5%,a specificity of 71.1%,and an accuracy of 80.2%;while the tandem test of MLR and NPAR predicted the occurrence of DME with a sensitivity of 46.4%,a specificity of 97.8%,and an accuracy of 69.3%.Conclusion Increased levels of MLR and NPAR are independent risk factors for the occurrence of DME and have certain predictive value for DME.The predictive value of combined MLR and NPAR test for DME is higher than that of separate test,and parallel experiment is more helpful for the early prediction of DME.
2.Construction and gene identification of myeloid-specific Spi1 knockout mice
Xuming WU ; Huihui WANG ; Xiangling ZHU ; Yuanyuan ZHOU ; Anqi WANG ; Huiru ZHANG ; Chong LIU ; Jiajie TU
Acta Universitatis Medicinalis Anhui 2024;59(3):413-417
Objective To construct myeloid-specific Spi1 gene knockout mice and analyze their genotypes,so as to provide animal model basis for the study of pathological mechanism of diseases and drug targets.Methods Ac-cording to the principle of CRISPR/Cas9 technology and Cre/LoxP system,sgRNA and Donor vectors were de-signed and constructed.The transcript of Exon 2(Exon 2)was used as the knockout region,and Loxp elements were placed on both sides of Exon 2.Cas9 protein,sgRNA and Donor vector were mixed and microinjected into the fertilized eggs of C57BL/6J mice,the fertilized eggs were transplanted into the uterus of C57BL/6J pregnant female mice,and F0 generation was obtained after 19~20 days.Positive F0 mice were mated with C57BL/6J mice to ob-tain stable F1 Spi1flox/+mice.Spi1flox/+mice of F1 generation were selfed to obtain Spi1flox/flox mice.Spi1flox/flox mated with Lyz2-Cre+mice to obtain Spi1flox/+/Lyz2-Cre+mice,and then mated with Spi1flox/flox,the Spi1flox/flox/Lyz2-Cre+mice were myeloid-specific Spi1 gene knockout(KO)mice.Spi1flox/flox/Lyz2-cre-mice were used as wild-type(WT)mice.DNA of WT and KO mice was extracted,and the genotypes were identified by agarose gel electro-phoresis after PCR amplification.Western blot was used to detect the expression of spleen focus forming virus provi-ral integration oncogene,Spi-1/purine rich box-1(PU.1)in immune cells of WT and KO mice.Results The results of PCR identification showed that the genotype of mice with only 220 bp amplified by flox primer was Spi1flox/flox homozygote,and the genotype of mice with 700 bp amplified by Lyz2-Cre primer was Lyz2-Cre+.Western blot showed that compared with WT group,the protein PU.1 was not expressed in bone marrow-derived macropha-ges(BMDMs)and peritoneal macrophages(PM)in KO group(P<0.01).There was no significant difference of statistics in the expression level of PU.1 in T cells between KO mice and WT mice.The results of PCR and West-ern blot showed that myeloid-specific Spi1 KO mice were successfully constructed.Conclusion The myeloid-spe-cific Spi1 gene KO mice are successfully constructed and identified,which provides animal model basis for further revealing the potential mechanism of PU.1 inimmune regulation.
3.Breeding and genotyping of T lymphocyte-conditional Spi1 knockout mice
Huihui WANG ; Xiangling ZHU ; Xuming WU ; Huiru ZHANG ; Yuanyuan ZHOU ; Anqi WANG ; Chong LIU ; Jiajie TU
Acta Universitatis Medicinalis Anhui 2024;59(4):595-599
Objective To breed and identify the T lymphocyte-conditional Spi1 knockout mice for the further in-vestgation of the specific role of Spi1-encoded protein PU.1.Methods The Lck-Cre mice were mated with Spi1flox/flox mice to obtain Lck-Cre×Spi1flox/flox mice(T lymphocyte-specific Spi1 knockout mice),and the genotype was determined by polymerase chain reaction(PCR)and agarose gel electrophoresis.Magnetic beads were used to sort out the splenic T lymphocytes,and the knockdown efficiency of PU.1 in T cells was detected by Western blot,quantitative real-time PCR(qPCR)and flow cytometry.Results The Lck-Cre×Spi1flox/flox mouse genotype was stably inherited.Compared with Spi1flox/flox mice,the expression level of PU.1 was significantly reduced in splenic T cells of Lck-Cre×Spi1flox/flox mice.Conclusion In this study,the T lymphocyte-specific Spi1 knockout mice was successfully constructed by applying Cre/LoxP system and CRISPR/Cas9 technology,which provided a reliable an-imal model for the subsequent experiments of the specific role of PU.1 in T cell-related diseases.
4.Construction and efficiency detection of Csf1r-CreERT2 R26REYFP reporter gene mouse based on Cre/Loxp system
Xiangling ZHU ; Xuming WU ; Huihui WANG ; Yuanyuan ZHOU ; Anqi WANG ; Huiru ZHANG ; Chong LIU ; Jiajie TU
Acta Universitatis Medicinalis Anhui 2024;59(7):1175-1180
Objective To construct Csf1r-CreERT2 R26REYFP reporter gene mice and assess the efficacy of Csf1r-CreERT2-mediated enhancement of CSF1R in CD45+cells labeled with yellow fluorescein protein EYFP.Methods Csf1r-CreERT2 mice were crossbred with R26REYFP homozygous mice,and Csf1r-CreERT2R26REYFP mice were identified through PCR and Western Blot analyses.Flow cytometry was employed to evaluate CSF1R tag-efficiency in CD45+cells across different mouse tissues following tamoxifen induction.Results Csf1r-CreERT2 R26REYFP reporter gene mice were acquired.In addition,it was found that Csf1r-CreERT2-mediated EYFP could effectively mark CSF1R in various tissues of mice and CD45+cells in different locations.Compared to the R26REYF P group,the highest labeling efficiency was observed in the brain tissue(P<0.001),the lowest in the thymus tissue(P<0.05),and no sig-nificant difference was observed in the spleen tissue.Conclusion Adult Csf1r-CreERT2 mice and R26REYFP mice are effective ways to obtain Csf1r-CreERT2 R26REYFP induced conditional fluorescence mice.Csf1r-CreERT2 can mediate EYFP to effectively trace CSF1R in CD45+cells in different parts of mice.
5.Effect of small-incision lenticule extraction with different corneal cap thicknesses on postoperative astigmatism and short-term visual quality of patients with myopic astigmatism
Xinli ZHANG ; Yang LIU ; Jian FU ; Xiangling LIU
International Eye Science 2024;24(11):1811-1815
AIM: To compare the effect of small-incision lenticule extraction(SMILE)with different corneal cap thicknesses on postoperative astigmatism and short-term visual quality of patients with myopic astigmatism.METHODS: A total of 54 patients(108 eyes)with myopic astigmatism who underwent SMILE from June 2020 to June 2022 in our hospital were selected for the prospective controlled study, and patients were randomly assigned into two groups, with 27 cases(54 eyes)each. The corneal cap thickness design was 110 μm for the group A and 120 μm for the group B, while other operation parameters were consistent. Additionally, the uncorrected visual acuity(UCVA), spherical equivalent(SE), stiffness parameter A1(SP-A1), visual quality and vector parameters at baseline, 1 d,1 wk and 1 mo after surgery were compared between two groups.RESULTS: There was a statistically significant difference in UCVA, SE, and SP-A1 between the two groups at various time points before and after surgery(all P<0.05), and UCVA in the group A was better than that in the group B at 1 d after surgery(P<0.05). There was no statistically significant difference in the results of astigmatism vector analysis between the two groups of patients(both P>0.05). The objective scattering index(OSI)of the group A was lower than that of the group B, while Strehl ratio(SR)of the group A was higher than that of the group B at 1 d after surgery(both P<0.05). There was no significant difference in modulation transfer function cutoff frequency(MTF cut off), contrast vision, visual symptoms and overall satisfaction, postoperative complications between the two groups(all P>0.05).CONCLUSION: SMILE procedures with both 110 μm and 120 μm corneal cap thicknesses are safe and effective in correcting myopic astigmatism without affecting postoperative SE, astigmatism, SP-A1 or contrast visual acuity. Whereas 110 μm corneal cap thickness results in faster early postoperative visual recovery and better early visual quality than 120 μm.
6.Expression and significance of SHP2 in retinal tissues of mice with oxygen-induced retinopathy
Jin CHEN ; Xiangling LIU ; Shaobo SU ; Gaoen MA
Journal of Xinxiang Medical College 2024;41(11):1016-1021
Objective To investigate the expression and significance of Src homology 2 domain-containing protein tyrosine phosphatase 2(SHP2)and phosphorylated SHP2(P-SHP2)in oxygen-induced retinopathy(OIR).Methods Twenty clean-grade C57BL/6J(B6)7-day-old neonatal mice were randomly divided into normal control group and OIR group,with 10 mice in each group.The mice in the normal control group and the nursing mother mice were fed together in a normal oxygen and room temperature environment.Mice in the OIR group were placed in an oxygen chamber with a temperature of 22-25 ℃,humidity of(60±10)%and oxygen volume fraction stable at(75±5)%for five days together with the nursing mother mice,and then transferred to a normal oxygen environment.At 12,14,17 days of age,three mice were selected from the normal con-trol group and the OIR group,respectively;and the expressions of SHP2 and P-SHP2 proteins in the retinal tissues of mice in the two groups were detected by Western blot.Two 17-day-old mice were randomly selected from the normal control group and the OIR group,respectively;and the retinal histopathology of mice was observed by hematoxylin-eosin staining.Results In the normal control group,the cell structure in all layers of the retinal tissue of 17-day-old mice was orderly,the inner limiting mem-brane and endothelial nucleus were intact,and the shape was normal.In the OIR group,the intercellular structure in all layers of the retinal tissue of 17-day-old mice was disordered,and the vascular endothelial nucleus was seen to break through the reti-nal inner limiting membrane.There was no statistically significant difference in the relative expression of SHP2 protein in the retinal tissues of mice at different ages in the normal control group(F=2.052,P>0.05).The relative expression level of P-SHP2 protein in the retinal tissues of mice at 14,17 days of age in the normal control group was significantly lower than that at 12 days of age(P<0.05),and the relative expression level of P-SHP2 protein in the retinal tissues of mice at 17 days of age was significantly lower than that at 14 days of age(P<0.05).The relative expression level of SHP2 protein in the retinal tissues of mice at 14 days of age in the OIR group was significantly higher than that at 12,17 days of age(P<0.05).There was no statistically significant difference in the relative expression level of SHP2 protein in the retinal tissues between 17-day-old mice and 12-day-old mice in the OIR group(P>0.05).The relative expression level of P-SHP2 protein in the retinal tissues of mice at 14 days of age in the OIR group was significantly higher than that at 12,17 days of age(P<0.05).There was no statistically significant difference in the relative expression level of P-SHP2 protein in the retinal tissues between 17-day-old mice and 12-day-old mice in the OIR group(P>0.05).The relative expression level of SHP2 protein in the retinal tissues of mice at 12,17 days of age in the OIR group was significantly lower than that in the normal control group(P<0.05).There was no statistically significant difference in the relative expression level of SHP2 protein in the retinal tissues of mice at 14 days of age between the two groups(P>0.05).The relative expression level of P-SHP2 protein in the retinal tissues of 12-day-old mice in the OIR group was significantly lower than that in the normal control group(P<0.05),while the relative expression level of P-SHP2 protein in the retinal tissues of 17-day-old mice was significantly higher than that in the normal control group(P<0.05).There was no statistically significant difference in the relative expression level of P-SHP2 protein in the retinal tissues of mice between the two groups at 14 days of age(P>0.05).Conclusion SHP2 and P-SHP2 are expressed with tem-poral fluctuations in OIR.Hypoxia may promote conformational changes of SHP2 in OIR mice,and play an active role in phos-phorylation;and participate in and promote the occurrence and development of OIR.
7.Safety and Effectiveness of Chloral Hydrate for Auditory Brainstem Response Tests in Children
Xiangling ZHANG ; Xinyi YAO ; Min LIU ; Youyou JIN ; Zhaoli MENG ; Haotian LIU ; Yun ZHENG
Journal of Audiology and Speech Pathology 2024;32(6):524-527
Objective To study the safety and effectiveness of chloral hydrate in children undergoing ABR tests.Methods From December 2015 to March 2022,5 513 children under the age of 12 were selected for ABR ex-amination in West China Hospital of Sichuan University,who received chloral hydrate sedation(dose of 30 mg/kg).Data on administration method(mixed or direct),sleep deprivation(yes or no),failure performance(such asfailure to sleep,insufficient sedation,superficial sleep),adverse events(vomiting,irritability,etc.)were retrospectively analyzed.Total sedation failure rate,sedation failure rates in different age groups(≤0.5 years,0.5~3 years,3~12 years)and adverse event rate were calculated.Results Among the 5 513 ABR tests,199(3.61%)failed seda-tion.The sedation failure rates in different age groups(≤0.5 years,0.5~3 years,3~12 years)were 3.03%,4.31%and 3.11%,respectively.In the sedation failure tests,insufficient sedation was found in 81.91%of the tests.The incidence of adverse events was 10.55%,with most commonly vomiting.Conclusion The sedation fail-ure rate and the incidence of adverse events of chloral hydrate at 30 mg/kg were relatively low,thus chloral hydrate can be considered safe and effective at this dose.
8.Stratified Treatment in Pediatric Anaplastic Large Cell Lymphoma: Result of a Prospective Open-Label Multiple-Institution Study
Tingting CHEN ; Chenggong ZENG ; Juan WANG ; Feifei SUN ; Junting HUANG ; Jia ZHU ; Suying LU ; Ning LIAO ; Xiaohong ZHANG ; Zaisheng CHEN ; Xiuli YUAN ; Zhen YANG ; Haixia GUO ; Liangchun YANG ; Chuan WEN ; Wenlin ZHANG ; Yang LI ; Xuequn LUO ; Zelin WU ; Lihua YANG ; Riyang LIU ; Mincui ZHENG ; Xiangling HE ; Xiaofei SUN ; Zijun ZHEN
Cancer Research and Treatment 2024;56(4):1252-1261
Purpose:
The risk stratification of pediatric anaplastic large cell lymphoma (ALCL) has not been standardized. In this study, new risk factors were included to establish a new risk stratification system for ALCL, and its feasibility in clinical practice was explored.
Materials and Methods:
On the basis of the non-Hodgkin’s lymphoma Berlin–Frankfurt–Munster 95 (NHL-BFM-95) protocol, patients with minimal disseminated disease (MDD), high-risk tumor site (multiple bone, skin, liver, and lung involvement), and small cell/lymphohistiocytic (SC/LH) pathological subtype were enrolled in risk stratification. Patients were treated with a modified NHL-BFM-95 protocol combined with an anaplastic lymphoma kinase inhibitor or vinblastine (VBL).
Results:
A total of 136 patients were enrolled in this study. The median age was 8.8 years. The 3-year event-free survival (EFS) and overall survival of the entire cohort were 77.7% (95% confidence interval [CI], 69.0% to 83.9%) and 92.3% (95% CI, 86.1% to 95.8%), respectively. The 3-year EFS rates of low-risk group (R1), intermediate-risk group (R2), and high-risk group (R3) patients were 100%, 89.5% (95% CI, 76.5% to 95.5%), and 67.9% (95% CI, 55.4% to 77.6%), respectively. The prognosis of patients with MDD (+), stage IV cancer, SC/LH lymphoma, and high-risk sites was poor, and the 3-year EFS rates were 45.3% (95% CI, 68.6% to 19.0%), 65.7% (95% CI, 47.6% to 78.9%), 55.7% (95% CI, 26.2% to 77.5%), and 70.7% (95% CI, 48.6% to 84.6%), respectively. At the end of follow-up, one of the five patients who received maintenance therapy with VBL relapsed, and seven patients receiving anaplastic lymphoma kinase inhibitor maintenance therapy did not experience relapse.
Conclusion
This study has confirmed the poor prognostic of MDD (+), high-risk site and SC/LH, but patients with SC/LH lymphoma and MDD (+) at diagnosis still need to receive better treatment (ClinicalTrials.gov number, NCT03971305).
9.Exploration and practice of the scientific and technological achievements translation at university affiliated hospitals
Xiangling QIAN ; Huanlong QIN ; Xinming JIA ; Jun YIN ; Guojun CAI ; Yiming WANG ; Rui LIU
Chinese Journal of Hospital Administration 2023;39(2):124-128
As an important element of medical and health sector innovation, the translation of scientific and technological achievements plays a key role in promoting their clinical application and meeting the medical needs of the people. The authors sorted out the problems in such translation at these affiliated hospitals in terms of " people", " finance", " material", and " system". Starting from 2017, the Tenth People′s Hospital Affiliated to Tongji University has explored such practices as establishing hospital-led clinical medical science and technology innovation parks and technology service limited companies. These practices aimed to address the issues of insufficient hospital scientific and technological innovation capabilities and the gap between the hospital′s operation mechanism to translate its scientific and technological achievements and the enterprises and the market. The clinical medical science and technology innovation park integrating administration, industry, education, research, medicine and application, has taken multiple measures to attract excellent research talents and projects from within and beyond the hospital, promote the implementation of innovative scientific research projects. The hospital also established a health industry mode with engagement of social capital from large enterprises. The Technology Services Co., Ltd. was based on the incubation and translation of hospital achievements, combining market and clinical needs, promoting multi-party cooperation between hospitals and external enterprises, improving the chain operation mechanism of hospital scientific and technological achievements translation work, and alleviating the problem of insufficient research pilot funds and productibility funds by means of hospital-led fundraising. The number of patent authorizations of hospitals had increased from 23 cases in 2018 to 105 in 2022, and the amount of patent conversion had increased from 2 million yuan in 2020 to 11 million yuan in 2022. It is recommended that affiliated hospitals of universities further improve the organizational structure of achievement translation, strengthen their professional talent teams, improve their operation mechanism of achievement translation, build a platform for medical school-enterprise cooperation, and improve the evaluation mechanism of translation assessment, in order to promote a virtuous cycle of hospital′s scientific and technological achievement translation work.
10.Construction of a chimeric antigen receptor CAR THP -1 cell line targeting HER2
Yizhao Chen ; Lihua Liu ; Xiangling Zhu ; Huihui Wang ; Xuming Wu ; Xuewen Tan ; Yilong Fang ; Haifeng Jiang ; Zhen Xu ; Wei Wei ; Jiajie Tu
Acta Universitatis Medicinalis Anhui 2023;58(3):352-357
Objective:
To obtain chimeric antigen receptor macrophages ( CAR-M) targeting HER2 stably transfected.
Methods :
CAR lentivirus vector targeting HER2 was constructed and infected with human monocytic leukemia cell line (THP-1) .CAR THP-1 cells with green fluorescent labeling were selected by sorting flow cytometry and continued to be cultured in vitro.The CAR THP-1 cells targeting HER2 were co-cultured with the endometrial cancer cell line Ishikawa with negative and positive HER2 expression,and their targeted phagocytosis of CAR-M to HER2 positive tumor cells was detected by imaging flow cytometry ,and the targeted phagocytosis efficiency of CAR-M to HER2 positive tumor cells was detected by flow cytometry.
Results :
CAR lentivirus infection with THP- 1 cells was less efficient ; After co-culture with cancer cells,flow cytometry and imaging flow cytometry showed that CAR THP-1 cells had enhanced phagocytosis of HER2 positive Ishikawa cells compared with the empty body group (P<0. 01) .
Conclusion
In this experiment,CAR THP-1 cell line targeting HER2 was established by constructing CAR lentivirus vector and transfecting THP-1 cells ,and it was proved that CAR THP-1 could phagocytize HER2 positive Ishikawa cells through specific targeting.


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