1.Non-classical 21-hydroxylase deficiency of a pedigree and related literature review
Xinling WANG ; Fangyu ZHAO ; Jimilanmu Maimaitiming ; Yuan CHEN ; Yanying GUO
Chinese Journal of Endocrinology and Metabolism 2019;35(10):838-842
The clinical data of a patient with non-classical 21-hydroxylase deficiency ( 21-OHD) were retrospectively analyzed. The CYP21A2 gene analysis was performed on the patient and his family members by PCR-DNA direct sequencing. It was found that the proband had a heterozygous mutation [ point mutation:p.Ile173Asn, p. ( Ile237Asn, Val238Glu, Met240Lys ), p. Val282Leu, p. Gln319Ter, p. Arg357Trp, insertion mutation: p.Leu308Phefs?6, deletion/insert mutation: p. Arg484Profs]. Among the members of the family, the patient's eldest sister and three paternal members all carried the p.Val282Leu heterozygous mutation, and the patient's second sister and two maternal members carried the same p. Val282Leu homozygous mutation and other compound heterozygous mutations just as the proband. The proband presented a non-classical phenotype with ultimately normal height and fertility. It is suggested that the potential phenotype of the disease is related to the residual activity of its allele, and there exists a good genotype-phenotype correlation.
2.Inflammatory bowel disease with thrombosis: a single center retrospective study
Jin LI ; Juan WEI ; Chunyan CHEN ; Li ZHAO ; Zhijian CHEN ; Lin WU ; Xingjiang WU ; Fangyu WANG ; Jiong LIU
Chinese Journal of Digestion 2019;39(1):31-34
Objective To analyze the clinical features and clinical significance of patients with inflammatory bowel disease (IBD) complicated by thrombosis.Methods From March 14th,2001 to February sixth 2017,at Nanjing General Hospital of Nanjing Military Command,27 IBD patients with thrombosis diagnosed by clinical symptoms,endoscopy,imaging and pathology were enrolled.During the same period,81 gender and age matched IBD patients without thrombosis were included in the control group.The basic data,information of IBD diagnosis and treatment and thrombotic events of patients were collected and analyzed.T-test and Chi-square test were performed for statistical analysis.Binary logistics regression was used for risk factors analysis.Results The mean age of diagnosis of IBD patients with thrombosis was (44.8 ± 15.8) years,which was higher than that of the IBD patients of control group ((36.0 ± 14.4) years),and the difference was statistically significant (t =2.69,P =0.008).Among 27 IBD patients with thrombosis,arterial thrombosis was 51.9% (14/27),deep venous thrombosis of the lower extremity veins was 29.6% (8/27),portal venous system involved was 11.1% (3/27),pulmonary embolism was 3.7% (1/27) and disseminated intravascular coagulation accounted for 7.4% (2/27).Nine patients (33.3%,9/27) underwent surgery six months before the diagnosis of thrombosis.The results of binary logistic regression indicated that the age of diagnosis and vascular catheterization were independent risk factors for thrombosis in IBD patients (odds ratio (OR) =1.04,95% confidence interval (CI) 1.01 to 1.07,P=0.01;0R=5.64,95% CI 1.39 to 22.96,P=0.02).After medicine treatment or surgery,81.5% (22/27) of the patients improved,9.1% (2/22) were worse and 13.6% (3/22) died.Conclusion Screening and prevention of thrombosis should be paid attention in IBD patients with a history of vascular catheterization,at active phase and older age when diagnosed.
3.Research progress of PARP inhibitors in cancers and their drug resistance
Wanwan YANG ; Fangyu YE ; Yujia WU ; Haochen WANG ; Li ZHAO
Journal of China Pharmaceutical University 2022;53(5):525-534
Poly ADP-ribose polymerase-1 (PARP-1) plays a vital role in organisms, including regulating repair of DNA, maintaining genome stability, regulating cell proliferation, differentiation, and death.At present, PARP inhibitors have been made some breakthrough in the treatment of breast cancer, ovarian cancer, prostate cancer and pancreatic cancer.However, PARP inhibitors have certain limitations in other malignant tumors and patients who are resistant to PARP-1 inhibitors.This article summarizes the research on PARP inhibitors in lung cancer, hepatocellular carcinoma, glioblastoma, leukemia and cervical cancer, and introduces the strategies of combining other anti-tumor drugs such as DNA repair inhibitors, immune checkpoint inhibitors, anti-angiogenic drugs and other chemotherapeutic drugs to solve their drug resistance, which provides some reference for the wide clinical application of PARP inhibitors in the future.
4.Distribution of food specific antibodies in patients with inflammatory bowel disease
Yulin YANG ; Chang LIU ; Zhao YANG ; Hui TAO ; Fangyu WANG
Chinese Journal of Digestion 2019;39(7):444-451
Objective To investigate the incidence of food intolerance in patients with inflammatory bowel disease (IBD) and to analyze the differential diagnostic value of intolerant food in Crohn 's disease (CD) and ulcerative colitis (UC) and its effects on the diseases.Methods From January 2017 to June 2018, at Department of Gastroenterology and Hepatology , Jinling Hospital, Medical School of Nanjing University /General Hospital of Eastern Theater Command , PLA, a total of 252 IBD (154 CD and 98 UC) patients were enrolled. In the same period 46 non-IBD patients were recruited.Allergic diseases were excluded.The food-specific IgG antibodies were detected by enzyme linked immunosorbent assay (ELISA) in all enrolled patients.The chi-square test, Kruskal-Wallis test and Spearman correlation analysis were performed for statistical analysis . Regression analysis was used to screen the risk factors .Results The total positive rates of serum IgG antibody of corn, rice, soybean, tomato and wheat in CD patients were 60.4% (93 /154), 57.8% (89 /154), 42.9%(66 /154), 68.2% (105 /154) and 19.5% (30 /154),respectively, which were higher than those in patients with UC (7.1%, 7 /98; 5.1%, 5 /98; 5.1%,5 /98; 16.3%, 16 /98 and 3.1%, 3 /98, respectively) and those of non-IBD patients (2.2%,1 /46; 2.2%,1/46; 0,0 /46; 0,0 /46 and 0,0/46, respectively), and the differences were statistically significant (χ2 =70.940,71.092,42.185,64.517,14.187;48.190,44.270, 29.424,66.029,10.542; all P <0.01).The total number of positive food related IgG antibodies had strong ability to discriminate CD, UC from non-IBD, the value of area under the curve (AUC) was 0.815 (95%confidence interval (CI) 0.762 to 0.869, P <0.01)and 0.824(95%CI 0.767 to 0.880,P <0.01).There was no correlation between the total number of IgG positive food and age in CD patients (P >0.05) and there was a negative correlation between the total number of IgG positive food and age in UC patients (rs =-0.376, P <0.01).The median number of total IgG positive food of patients with lesions involving the terminal ileum (L1) and ileocolon (L3) was two and four, respectively, and the difference was statistically significant (the statistic was 11.717,P =0.002).The median number of total IgG positive food of UC patients with rectal lesions (E1) and extensive colon lesions (E3) was zero and one, respectively, and the difference was statistically significant (the statistic was 7.191,P =0.022).In addition, positive IgG shrimp and soybean were risk factors of CD patients combined with extra-intestinal manifestations and low body mass index (odd ratio (OR) =24.558, 95%CI 2.243 to 268.936; OR =2.253, 95% CI 1.048 to 4.841; both P <0.05, respectively ). Conclusions CD patients are more susceptible to food intolerance .The number of intolerant foods have differential diagnostic value in CD, UC and non-IBD.The larger the lesion of IBD patients, the more common the food intolerance.IgG antibody positive food positive, may affect extra-intestinal manifestations and nutritional status of CD patients .
5.Effects of RARS2 on cell proliferation, invasion, migration and chemotherapy resistance of pancreatic cancer
Yueze LIU ; Jiangdong QIU ; Gang YANG ; Fangyu ZHAO ; Taiping ZHANG
Chinese Journal of Hepatobiliary Surgery 2022;28(5):368-372
Objective:To investigate the effects of mitochondrial arginyl-tRNA synthase (RARS2) on cell proliferation, invasion, migration and chemotherapy resistance of pancreatic cancer.Methods:Human pancreatic cancer cell lines AsPC-1 and PANC-1 were divided into negative control group, RARS2 interference group-1, RARS2 interference group-2, RARS2 overexpression control group and RARS2 overexpression group. Cell proliferation and sensitivity to gemcitabine were detected by CCK-8 assay, and cell invasion and migration were detected by Transwell assay. Western blot was used to detect the expression of RARS2 under different concentrations and different times of gemcitabine treatment. Western blot and PCR were used to detect the expression of RARS2 in gemcitabine-resistant AsPC cell.Results:Inhibition of RARS2 expression in AsPC-1 and PANC-1 cells significantly inhibited cell proliferation and enhanced sensitivity of gemcitabine to chemotherapy. Overexpression of RARS2 enhanced cell proliferation and decreased sensitivity to gemcitabine. In AsPC-1 cells, the number of migrated cells (100×) in negative control group, RARS2 interference group-1, RARS2 interference group-2, RARS2 overexpression control group and RARS2 overexpression group were (586.7±37.4) cells/field, (195.7±18.6) cells/field, (237.0±17.1) cells/field, (157.7±19.1) cells/field, (456.0±23.1) cells/field, the number of invasive cells were (87.7±13.2) cells/field, (24.7±6.5) cells/field, (31.7±6.1) cells/field, (29.3±4.5) cells/field, (94.3±9.3) cells/field, respectively. The migration and invasion ability of cells were decreased after the expression of RARS2 was decreased, and the migration and invasion ability of cells were enhanced after the expression of RARS2 was increased. PCR and Western blot assay showed that RARS2 expression in the gemcitabine-resistant AsPC-1 was higher than that in the common cell line. In AsPC-1 cells, the expression of RARS2 increased with increasing gemcitabine concentration and treatment time.Conclusion:RARS2 promotes cell proliferation, invasion, migration and chemoresistance of pancreatic cancer, and expression of RARS2 is positively correlated with gemcitabine concentration and treatment time.
6.Toddalia asiatica attnuates cerebral ischemia-reperfusion injury in rats through modulating M1/M2 polarization of microglia
Jianhong GAO ; Gang WANG ; Dan YANG ; Fangyu ZHAO ; Yiduo HE ; Xian-Bing CHEN
Chinese Journal of Pathophysiology 2023;39(12):2133-2140
AIM:Observation of neuroprotective effects of Toddalia asiatica(TA)on cerebral ischemia-reper-fusion injury(CIRI)in rats by investigating the effects and mechanisms of drugs on the polarization of microglia M1/M2 subtype and the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)path-way.METHODS:The modified thread occlusion method was used to establish a rat model of CIRI,and the rats were ran-domly divided into the model group,Toddalia asiatica(1.08 g/kg)group,donepezil hydrochloride(0.45 mg/kg)group,and sham group,with 16 rats in each group.Based on the assessment of neurofunctional changes in each group of rats,HE and Nissl staining were used to observe the pathological changes in brain tissue.TUNEL staining was performed to de-tect neuronal apoptosis,immunohistochemistry was used to detect the expression of M1 microglia marker ionized calcium-binding adapter molecule 1(Iba1),M2 microglia marker arginase 1(Arg1),and TLR4 localization.Western blot was used to detect the expression of microglia polarization proteins and proteins related to TLR4/MyD88/NF-κB pathway in the hippocampus region.RESULTS:Compared with sham group,the model group rats had higher neurological function scores(P<0.01),and neuronal arrangement in the hippocampus and cortex was loose and disordered,Nissl bodies de-creased,and neuronal apoptosis increased.The numbers of M1 microglia marker Iba1-,M2 microglia marker Arg1-,and TLR4-positive cells were significantly increased.In addition,the protein levels of TLR4,MyD88,p-NF-κB p65,NF-κB p65,p-NF-κB inhibitory factor(p-IκB),Iba1,interleukin-6(IL-6),and Arg1 in the hippocampus were elevated(P<0.05),while IL-4 and IL-10 expression were decreased(P<0.01).Compared with model group,the Toddalia asiatica group and Donepezil hydrochloride group showed increased protein expression of Arg1,IL-4 and IL-10(P<0.05),while the other indicators were decreased(P<0.05,P<0.01).CONCLUSION:Toddalia asiatica possesses neuroprotective effects on CIRI rats,which may be attributed to its ability to regulate M1/M2 polarization and inhibit the TLR4/MyD88/NF-κB-mediated inflammatory pathway.
7.Study on the Effects of Ginseng Root Extract on Autophagy ,Proliferation and Cytokine Secretion of Mice Splenic Lymphocytes and Its Mechanism
Fangyu LI ; Bin QI ; Shuai BIAN ; Yue ZHAO ; Shuyan LU ; Jiawen WANG ; Daqing ZHAO
China Pharmacy 2020;31(21):2597-2602
OBJECTIVE:To study the effects of gin seng root extract on autophagy ,proliferation and cytokine of splenic lymphocyte of mice ,and to study its mechanism. METHODS :The splenic lymphocyte of mice were divided into blank control group,positive control group (concanavalin A ,5 μg/mL),different concentration of ginseng root extract groups (32,125,500 μg/mL,by lyophilized powder ). After 48 h of culture with the corresponding medicine ,acridine orange staining method was used to detect autophagy of splenic lymphocyte ;CCK-8 assay was used to detect the cell proliferation ;ELISA assay was used to determine the levels of IL- 4,IL-6 and TNF-α in cell culture;Western blotting method was used to detect the expression of LC 3B and Beclin- 1,as well as the phosphorylation of AMPK ,AKT and mTOR. RESULTS :Compared with blank control group ,32, 125,500 μg/mL ginseng root extract could increase the number of acidic autophagosomes in splenic lymphocytes of mice(P<0.05 or P<0.01);125,500 μg/mL Ginseng root extract could significantly enhance the survival rate of splenic lymphocytes,the levels of IL- 4,IL-6 and TNF-α and the transformation of LC3BⅠ to LC 3BⅡ,significantly increased the protein expression of Beclin- 1, quinacrine cationic liposomes for treating non-small cell @163.com lung cancer[J]. J Drug Target ,2015,23(3):232-243. the phosphorylation of AMPK protein ,and significantly reduced the phosphorylation level of AKT and mTOR proteins ,with statistical significance (P<0.05 or P<0.01). CONCLUSIONS :Ginseng root extract can induce autophagy of mice splenic lymphocytes,activate the activity of splenic lymphocytes ,regulate the secretion of cytokine by activating AMPK and inhibiting the activation of AKT which can inhibit the activity of mTOR ,so as to exert immune enhancement effect.
8.Material Basis and Pharmacological Action of Tongxie Yaofang: A Review
Qingyu ZHAO ; Miao ZHANG ; Wenjing ZHAO ; Yiming WANG ; Fangyu SUI ; Zihe GUAN ; Jianwei WANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(8):237-245
Tongxie Yaofang, also known as Baizhu Shaoyaosan, was first recorded in Danxi's Experiential Therapy (《丹溪心法》) by ZHU Danxi in the Yuan dynasty. It is composed of Atractylodis Macrocephalae Rhizoma, Paeoniae Radix Alba, Citri Reticulatae Pericarpium, and Saposhnikoviae Radix, and serves as the representative prescription for the treatment of painful diarrhea. It has the functions of tonifying the spleen, emolliating the liver, relieving pain, and checking diarrhea, and is mainly used in the treatment of gastrointestinal diseases such as irritable bowel syndrome (IBS) and ulcerative colitis (UC). In addition, it is effective in treating gastrointestinal disorders with mental and psychological abnormalities, as well as obstinate anorexia in children, depression syndrome, and respiratory diseases. Experimental research and clinical practice have shown that Tongxie Yaofang has multi-component, multi-pathway, and multi-target characteristics in the treatment of diseases. The mechanism of Tongxie Yaofang in treating diseases is mainly attributed to anti-inflammation, immune function regulation, intestinal hypersensitivity improvement, emotion regulation, etc. Monoterpene glycosides, flavonoids, chromones, lactones, and other components contained play an important therapeutic role. The research on the systems biology of Tongxie Yaofang, such as metabolomics, proteomics, and network pharmacology, provides a scientific basis for clarifying its mechanism of action and expanding its clinical application. However, there are still some problems to be solved, such as difficulty in combining diseases and syndromes and lack of in-depth systematic research. Through the retrieval and collation of relevant literature, this paper systematically reviewed the material basis, pharmacological effects, and systems biology research of Tongxie Yaofang, aiming to lay a foundation for in-depth research on its mechanism in treating diseases and rational application in clinical practice.
9. The long coding RNA GSTM3TV2 acts an oncogene to promote chemoresistance in pancreatic cancer
Guangbing XIONG ; Gang YANG ; Mengyu FENG ; Fangyu ZHAO ; Lianfang ZHENG ; Taiping ZHANG ; Yupei ZHAO
Chinese Journal of Surgery 2019;57(9):691-697
Objectives:
To examine the expression of the long coding RNA GSTM3TV2 in pancreatic cancer tissues and to examine its role and mechanism in chemoresistance of pancreatic cancer cells.
Methods:
The expression of lncRNA GSTM3TV2 in 15 pancreatic cancer specimens and corresponding adjacent to cancer tissue samples diagnosed by Department of Pathology, Peking Union Medical College Hospital was detected by real-time PCR.And the expressions of GSTM3TV2 in pancreatic cancer cell AsPC-1, BxPC-3, MIAPaCa-2, PanC-1, SU86.86, T3M4, and chemoresistant cells AsPC-1/GR and MIAPaCa-2/GR, and human pancreatic nestin-expressing cells hTERT-HPNE were detected. Pancreatic cancer cell lines were transfected with GSTM3TV2-pcDNA3.1(+)in order to get cells with GSTM3TV2 overexpression.GSTM3TV2-siRNA was transfected into pancreatic cancer cells to knock down GSTM3TV2. The cell chemoresistance was measured by CCK-8 and flow cytometry assay when incubated with nab-paclitaxel. At the same time, subcutaneous xenograft tumor models were established in nude mice to observe the effect of GSTM3TV2 on chemoresistance of tumor growth in nude mice.Western blot assay was also performed to detect the molecular mechanism of chemoresistance of GSTM3TV2.
Results:
Comparing toadjacent tissues(0.084±0.019), GSTM3TV2 expression was significantly upregulated in the pancreatic cancer tissues(0.493±0.084) (
10.Resolving the lineage relationship between malignant cells and vascular cells in glioblastomas.
Fangyu WANG ; Xuan LIU ; Shaowen LI ; Chen ZHAO ; Yumei SUN ; Kuan TIAN ; Junbao WANG ; Wei LI ; Lichao XU ; Jing JING ; Juan WANG ; Sylvia M EVANS ; Zhiqiang LI ; Ying LIU ; Yan ZHOU
Protein & Cell 2023;14(2):105-122
Glioblastoma multiforme (GBM), a highly malignant and heterogeneous brain tumor, contains various types of tumor and non-tumor cells. Whether GBM cells can trans-differentiate into non-neural cell types, including mural cells or endothelial cells (ECs), to support tumor growth and invasion remains controversial. Here we generated two genetic GBM models de novo in immunocompetent mouse brains, mimicking essential pathological and molecular features of human GBMs. Lineage-tracing and transplantation studies demonstrated that, although blood vessels in GBM brains underwent drastic remodeling, evidence of trans-differentiation of GBM cells into vascular cells was barely detected. Intriguingly, GBM cells could promiscuously express markers for mural cells during gliomagenesis. Furthermore, single-cell RNA sequencing showed that patterns of copy number variations (CNVs) of mural cells and ECs were distinct from those of GBM cells, indicating discrete origins of GBM cells and vascular components. Importantly, single-cell CNV analysis of human GBM specimens also suggested that GBM cells and vascular cells are likely separate lineages. Rather than expansion owing to trans-differentiation, vascular cell expanded by proliferation during tumorigenesis. Therefore, cross-lineage trans-differentiation of GBM cells is very unlikely to occur during gliomagenesis. Our findings advance understanding of cell lineage dynamics during gliomagenesis, and have implications for targeted treatment of GBMs.
Mice
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Animals
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Humans
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Glioblastoma/pathology*
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Endothelial Cells/pathology*
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DNA Copy Number Variations
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Brain/metabolism*
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Brain Neoplasms/pathology*