1.siRNA AGAINST SURVIVIN SUPPRESSES THE PROLIFERATION OF PANCREATIC CANCER CELL PC-2 AND INDUCES ITS APOPTOSIS
Haitao GUAN ; Xinghuan XUE ; Xijing WANG ; Ang LI ; Zhaoyin QIN
Journal of Pharmaceutical Analysis 2006;18(1):78-82
Objective Blocking the expression of Survivin with siRNA, and the effects of suppling the proliferation of PC-2 cell and inducing its apoptosis were investigated. Methods Constructed the siRNA against Survivin plasmid expression vector and transfected it into PC-2 cell with lipofectamineTM 2000, the changes of Survivin expression were detected by semi-quantitive RT-PCR and immunohistochemical method, The effect of suppressing the proliferation of PC-2 cell was detected by the method of MTT; the effect of inducing PC-2 cell apoptosis was detected by flow cytometry. Results The sequence specific siRNA can effectively block the expression of Survivin both at themRNA and protein levels, the expression inhibition ratio was 81.25% at mRNA level and 74. 24% at protein level;blocking the expression of Survivin can suppress the proliferation of PC-2 cell significantly, 24, 48 hours after the cell was reseeded, the proliferation inhibition ratio was 28. 00% and 33. 38% respectively; 24, 48 hours after the transfection, 8.46 % and 7.53 % cells were induced to apoptosis respectively. Conclusion The siRNA against Survivin plasmid expression vector constructed in the study can blocking the expression of Survivin in PC-2 cell effectively and specifically; blocking the expression of Survivin can signiilcantly suppress the proliferation of PC-2 cell and induce a certain degree cells apoptosis; RNAi against Survivin is of a certain value in the gene therapy of pancreatic cancer.
2.Comparison of mechanical procedure and enzymatic digestion methods in isolating spermatogenic cells from rat testes
Shilin LI ; Xinghuan WANG ; Zhonghua YANG ; Xiaoyong PU ; Weicheng GAO ;
Chinese Journal of Tissue Engineering Research 2007;0(50):-
0.05).But the cell purity and the survival rate by mechanical procedure were significantly higher than that by combination of enzymatic digestion(P
3.Expressions and significance of two kinds of vascular endothelial growth factors of different origin in adrenocortical lesions of primary aldosteronism
Wen CHEN ; Zuojie LUO ; Yingfen QIN ; Xinghuan LIANG ; Decheng LU ; Jing XIAN ; Li JIANG
Chinese Journal of Endocrinology and Metabolism 2011;27(1):9-13
Objective To investigate the expression and significance of the endocrine gland-derived vascular endothelial growth factor (EG-VEGF) and vascular endothelial growth factor(VEGF) in adrenocortical lesions of primary aldosteronism. Methods The expressions of EG-VEGF, and VEGF were detected by immunohistochemistry and real-time fluorescence quantitative PCR in samples of 18 cases of adrenocortical adenoma, 6 adrenocortical hyperplasia, and 8 normal adrenal cortex. The correlation between the expressions of EG-VEGF, VEGF, and clinicopathological parameters was analyzed. Results The expression of EG-VEGF or VEGF in adrenocortical adenomas was higher than that in adrenocortical hyperplasia or normal adrenal cortex ( all P<0. 05 ), and the expression of EG-VEGF or VEGF between adrenocortical hyperplasia samples and normal adrenal cortex samples was indistinctive. There was no statistically significant correlation between EG-VEGF or VEGF expression and sex, age, blood pressure, serum potassium, plasma renin activity, except in case of serum aldosterone( P<0.05 ). A positive correlation between EG-VEGF and VEGF ( P<0. 01 ) was found. Conclusions EG-VEGF and VEGF may play a significant role in the formation and development of adrenocortical tumors in primary aldosteronism.
4.Clinical features and genetic analysis of a patient with type 2 neurofibromatosis manifested as oculomotor nerve palsy.
Xinghuan DING ; Bo LIANG ; Tingyu LIANG ; Jingjing LI ; Fang WANG ; Enshan FENG
Chinese Journal of Medical Genetics 2023;40(7):851-855
OBJECTIVE:
To report on a rare case of Neurofibromatosis type 2 (NF2) manifesting as oculomotor nerve palsy and explore its genetic basis.
METHODS:
A patient with NF2 who had presented at Beijing Ditan Hospital Affiliated to Capital Medical University on July 10, 2021 was selected as the study subject. Cranial and spinal cord magnetic resonance imaging (MRI) was carried out on the patient and his parents. Peripheral blood samples were collected and subjected to whole exome sequencing. Candidate variant was verified by Sanger sequencing.
RESULTS:
MRI revealed bilateral vestibular Schwannomas, bilateral cavernous sinus meningiomas, popliteal neurogenic tumors, and multiple subcutaneous nodules in the patient. DNA sequencing revealed that he has harbored a de novo nonsense variant of the NF2 gene, namely c.757A>T, which has replaced a codon (AAG) encoding lysine (K) at position 253 with a stop codon (TAG). This has resulted in removal of the Merlin protein encoded by the NF2 gene from position 253 onwards. The variant was not found in public databases. Bioinformatic analysis suggested that the corresponding amino acid is highly conserved. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was rated as pathogenic (PVS1+PS2+PM2_Supporting+PP3+PP4).
CONCLUSION
The heterozygous nonsense variant c.757A>T (p.K253*) of the NF2 gene probably underlay the disease in this patient with an early onset, atypical but severe phenotype.
Male
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Humans
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Neurofibromatosis 2/genetics*
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Genes, Neurofibromatosis 2
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Oculomotor Nerve Diseases/genetics*
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Computational Biology
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Genomics
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Mutation
5.Islet β-cell function in patients with gout and hyperuricemia accompanied by increased 1h postload plasma glucose
Jie ZHANG ; Guoran XIE ; Xinghuan LIANG ; Jia ZHOU ; Jing XIAN ; Song HUANG ; Li LI ; Yingfen QIN
Chinese Journal of Endocrinology and Metabolism 2018;34(4):300-303
All of 143 patients with gout or hyperuricemia were divided into type 2 diabetes(n=43), impaired glucose regulation(n=45),and normal glucose tolerance(n=55)groups. Moreover,a cut point of 8.6 mmol/L in one hour postload plasma glucose(1hPG)of oral glucose tolerance test was used to sub-divide the normal glucose tolerance group into 1hPG≥8.6 mmol/L(n=30)and 1hPG<8.6 mmol/L(n=25)groups. The first-and second-phase insulin secretion indexes were compared among four groups. The results showed that there was no statistical difference in the second-phase insulin secretion index among impaired glucose regulation,1hPG≥8.6 mmol/L,and 1hPG<8.6 mmol/L groups(P>0.05). The first-phase insulin secretion index revealed no significant difference between impaired glucose regulation and 1hPG≥8.6 mmol/L groups(P>0.05),but obviously decreased in these two groups compared with 1hPG<8.6 mmol/L group(P<0.05). The modified β cell function indexes were gradually decreased in 1hPG<8.6 mmol/L,1hPG≥8.6 mmol/L, and impaired glucose regulation groups(P<0.05). These results suggest that when 1hPG of the patients with gout and hyperuricemia is over 8.6 mmol/L,the first-phase insulin secretion will be impaired.
6.Correlation of neutrophil extracellular traps with T cells differentiation in Hashimoto′s thyroiditis mice and vitamin D′s relief effect
Xizhen WU ; Haiyan YANG ; Shien FU ; Yuping LIU ; Li LI ; Xinghuan LIANG ; Yingfen QIN ; Faquan LIN ; Zuojie LUO
Chinese Journal of Endocrinology and Metabolism 2023;39(2):149-155
Objective:To investigate correlation between neutrophil extracellular traps(NETs) formation and T cell subsets in mice with experimental autoimmune thyroiditis(EAT) and the impact of active vitamin D intervention.Methods:Six-week-old female BALB/c mice were randomly divided into Control group, EAT group and 1, 25 dihydroxy vitamin D 3[1, 25(OH) 2D 3] treatment group(VitD group; n=6/group). HE staining was used to observe thyroid pathology. Plasma thyroglobulin antibody(TGAb), thyroid peroxidase antibody(TPOAb), and 1, 25(OH) 2D 3 were measured by ELISA. Peripheral NETs formation, Th1, Th2, and Th17 cell ratio from spleen were measured by flow cytometry. Correlation between NETs formation rate and Th1, Th2, and Th17 cell ratio was analyzed. Results:Compared with Control group, mice in EAT group had significantly increased thyroid inflammation scores, thyroiditis morbidity, TPOAb, TGAb levels, NETs formation rate, Th2(CD4 + IL-4 + or CD4 + IL-13 + )and Th17 cell proportions( P were <0.001, 0.002, 0.007, <0.001, <0.001, 0.003, 0.001, and 0.002, respectively), and significant decreased 1, 25(OH) 2D 3, Th1 cell proportions, Th1/Th2(CD4 + IL-4 + ), Th1/Th2(CD4 + IL-13 + ), and Th1/Th17 ratios( P were 0.010, 0.018, 0.010, 0.005, and 0.007, respectively). Compared with the EAT group, the VitD group had lower thyroid inflammation scores, TPOAb, TGAb levels, NETs formation rate, Th2(CD4 + IL-4 + or CD4 + IL-13 + ) and Th17 cell proportions( P were 0.044, 0.007, <0.001, 0.001, 0.014, 0.008, and 0.001, respectively), and significant higher Th1 cell ratio, Th1/Th2(CD4 + IL-13 + ) and Th1/Th17 ratio( P were 0.011, 0.009, and 0.003, respectively). The Th1/Th2(CD4 + IL-4 + ) was not significantly increased in VitD group compared with EAT group( P=0.174). NETs formation rate was positively correlated with Th2(CD4 + IL-4 + or CD4 + IL-13 + ) and Th17 cell proportion( r were 0.65, 0.59, and 0.61; and P were 0.004, 0.010, and 0.007, respectively), but not with Th1 cell proportion( r=-0.47, P=0.051). Conclusion:EAT mice were more prone to NETs formation. Active vitamin D may relieve immune imbalance with increased Th2 and Th17 cell ratio and decreased Th1 cell ratio by reducing the formation of NETs in EAT mice. Vitamin D played the protective role in thyroid by reducing thyroid pathological damage and thyroid autoantibody levels, and relived overall lymphocyte imbalance.
7.Assessment of bleeding risk for early anticoagulation after endoscopic cyanoacrylate injection in cirrhosis patients with gastric varices and portal vein thrombosis
Huishan WANG ; Ye FANG ; Sitao YE ; Xinghuan LI ; Xiaoquan HUANG ; Jian WANG ; Lili MA ; Shiyao CHEN
Chinese Journal of Clinical Medicine 2024;31(3):361-366
Objective To explore the effects of anticoagulation treatment to postoperative bleeding events in liver cirrhosis patients with gastric varices and portal vein thrombosis.Methods Patients diagnosed with portal vein thrombosis and treated with endoscopic cyanoacrylate injection at Zhongshan Hospital,Fudan University due to gastric variceal bleeding from January 2023 to December 2023 were included.Clinical data of patients were collected,and patients were divided into anticoagulant group and non-anticoagulant group based on whether anticoagulant treatment was performed within 48 h after treatment.Re-bleeding in patients was evaluated in 6 weeks of follow-up.Cox regression was used for univariate and multivariate analysis of re-bleeding within 6 weeks after treatment.Results A total of 160 patients were included,of whom 65 patients received anticoagulation treatment within 48 h after endoscopic cyanoacrylate injection.There were no statistically significant differences in gender,etiology of liver cirrhosis,dosage of cyanoacrylate and sclerosing agents,and Child-Pugh grading between the two groups.There was no statistically significant difference in re-bleeding rate within 6 weeks after treatment between the two groups(1.54%vs 1.05%,P=0.795).Multivariate Cox regression analysis showed that the large amount of cyanoacrylate was a risk factor for re-bleeding within 6 weeks after endoscopic treatment(HR=5.862,P=0.015).Conclusions For patients with liver cirrhosis,gastric varices,and portal vein thrombosis,who receive endoscopic cyanoacrylate injection,early anticoagulation does not increase the risk of re-bleeding after treatment,while a large amount of cyanoacrylate injection may be a risk factor for re-bleeding.However,sample should be increased to verify.
8.Predictive value of multimodal ultrasound for upper urinary tract damage in children with neurogenic bladder
Junkui WANG ; Miao WANG ; Zikai LI ; Qinghua QI ; Yibo WEN ; Zhibin WU ; Xinghuan YANG ; Jianguo WEN
Chinese Journal of Applied Clinical Pediatrics 2024;39(12):911-916
Objective:To explore the predictive value of multimodal ultrasound (MMU) for upper urinary tract damage (UUTD) in children with neurogenic bladder (NB).Methods:This was a case-series study.From January 2022 to December 2023, 87 children with NB admitted to the First Affiliated Hospital of Zhengzhou University were examined by MMU.During the filling of bladder, vesical volume (VV), bladder wall thickness (BWT), shear wave velocity (SWV) of the anterior wall, resistance index (RI), and vascularization index (VI) were measured.After the emptying of bladder, VV and anterior wall SWV were measured, and ultrasound bladder compliance (△C) was calculated.The anterior posterior diameter (APD) of the renal pelvis and ureteral diameter (UD) were also measured.According to the upper/lower urinary tract dysfunction classification criteria, NB children were divided into a UUTD group and a non-UUTD (NUUTD) group.The differences in clinical data and related examinations between the 2 groups were analyzed to screen out independent risk factors, and an early warning model was established based on these factors.The prediction efficiency of the model and the urodynamic study (UDS) for UUTD was compared.Results:(1) There were 47 children in the UUTD group and 40 children in the NUUTD group.There was no significant difference in gender, age and body mass index between the 2 groups (all P>0.05).(2) In the UUTD group, the total glomerular filtration rate (tGFR) was (70.45±16.17) mL/min, the incidence of hydronephrosis was 38.30%, and the incidence of ureteral dilatation was 23.40%.No morphological changes were found in the imaging examination of the urinary system in the NUUTD group, and its tGFR was (100.55±16.27) mL/min.There was a significant difference in tGFR between the 2 groups ( P<0.05).(3) The filling VV, emptying VV, mean BWT, filling SWV, emptying SWV, VI, mean RI, △C, maximum cystometric capacity (MCC), maximum detrusor pressure during filling (Pdet.max), bladder compliance (BC), and detrusor leak point pressure (DLPP) in the NUUTD group were (218.43±87.53) mL, (14.62±6.14) mL, (3.08±0.65) mm, (2.64±0.54) m/s, (1.88±0.41) m/s, (6.20±1.04)%, 0.68±0.04, (147.58±49.18) mm 2·s, (309.50±66.54) mL, (59.83±19.79) cmH 2O(1 cmH 2O=0.098 kPa), (25.80±10.34) mL/cmH 2O, and (34.00±6.16) cmH 2O, respectively.Compared with the NUUTD group, the UUTD group showed decreased filling VV [(167.21±85.63) mL], △C [(78.49±31.86) mm 2·s], VI [(5.01±0.81) %], MCC [(255.32±75.10) mL], and BC [(12.57±6.44) mL/cmH 2O], and increased emptying VV [(19.50±7.65) mL], mean BWT [(4.02±0.82) mm], filling SWV [(3.99±1.07) m/s], emptying SWV [(2.15±0.35) m/s], mean RI (0.70±0.08), Pdet.max [(75.94±26.23) cmH 2O], and DLPP [(48.13±12.61) cmH 2O] (all P<0.05).(4) The decreased BC ( OR=0.841, 95% CI: 0.562-1.256, P=0.045), △C ( OR=0.427, 95% CI: 0.202-0.904, P=0.026) and VI ( OR=0.461, 95% CI: 0.091-2.325, P=0.010) and the increased DLPP ( OR=1.139, 95% CI: 0.894-1.451, P=0.040), filling SWV ( OR=1.895, 95% CI: 1.082-3.321, P=0.007) and mean BWT ( OR=1.191, 95% CI: 0.850-1.669, P=0.025) were independent risk factors for UUTD.Among MMU parameters, filling SWV had the highest prediction efficiency for UUTD, with a threshold of 3.33 m/s, sensitivity of 72.34% and specificity of 92.50%. Conclusions:MMU can well predict the occurrence of UUTD in children with NB, and filling SWV has the highest prediction efficiency.
9.Changes of iron metabolism in patients with hepatitis B cirrhosis complicated with esophageal and gastric varices and portal vein thrombosis
Sitao YE ; Yingjie AI ; Xinghuan LI ; Ye FANG ; Siyu JIANG ; Xiaoquan HUANG ; Shiyao CHEN
Journal of Chinese Physician 2024;26(4):489-493
Objective:To explore the changes of iron metabolism in patients with hepatitis B cirrhosis and esophageal and gastric varices complicated with portal vein thrombosis.Methods:This study was a cross-sectional study. 253 patients with hepatitis B cirrhosis with esophageal and gastric varices who were hospitalized in the Zhongshan Hospital, Fudan University from January 1, 2020 to December 31, 2021 were included in this study. They were divided into portal vein thrombosis group ( n=57) and non portal vein thrombosis group ( n=196) according to the presence or absence of portal vein thrombosis. The iron metabolism characteristics of the two groups were compared, and subgroups were analyzed according to the presence or absence of ascites, platelet count level, D-dimer level, and Child grade. The factors related to portal vein thrombosis were screened through multivariate logistic regression analysis. Results:The ratio of Child pugh B/C, ascites, D-dimer and platelet count in patients with hepatitis B cirrhosis and esophageal and gastric varices complicated with portal vein thrombosis was higher (all P<0.05). Patients with portal vein thrombosis had higher levels of soluble transferrin receptor [2.4(1.8, 3.6)mg/L vs 1.8(1.3, 2.7)mg/L, P=0.006], and lower levels of ferritin [33.1(18.9, 63.3)ng/ml vs 57.7(19.4, 142.5)ng/ml, P=0.038]. Layered analysis showed that ascites, platelet count levels, D-dimer levels, and Child-pugh grade did not affect the negative correlation trend between ferritin and portal vein thrombosis, and the positive correlation trend between soluble transferrin receptors and portal vein thrombosis. Moreover, soluble transferrin receptors showed a statistically significant positive correlation with portal vein thrombosis in the absence of ascites, low D-dimer levels, and Child-pugh grade A. Multivariate analysis suggested that after weighing Child-pugh grading, platelet count, and D-dimer levels, ferritin ( OR=0.943, 95% CI: 0.904-0.983, P=0.006) and soluble transferrin receptor ( OR=1.034, 95% CI: 0.001-1.067, P=0.044) were independently associated with portal vein thrombosis. Conclusions:In patients with hepatitis B cirrhosis with esophageal and gastric varices, the characteristics of iron metabolism in patients with portal vein thrombosis are different from those in patients without thrombosis, with higher levels of soluble transferrin receptor and lower levels of ferritin.
10.Application of holographic image navigation in urological laparoscopic and robotic surgery
Gang ZHU ; Jinchun XING ; Guobin WENG ; Zhiquan HU ; Ningchen LI ; He ZHU ; Pingsheng GAO ; Zhihua WANG ; Weizhi ZHU ; Kai ZHANG ; Hongbo LI ; Zhun WU ; Rui ZHU ; Xifeng WEI ; Yanan WANG ; Qun XIE ; Bing FU ; Xinghuan WANG ; Lin QI ; Xin YAO ; Tiejun PAN ; Delin WANG ; Nan LIU ; Jianguang QIU ; Jianggen YANG ; Bao ZHANG ; Zhuowei LIU ; Hui HAN ; Gang LI ; Bin ZHANG ; Manli NA ; Jingjing LU ; Lei WANG ; Zichen ZHAO ; Yanqun NA
Chinese Journal of Urology 2020;41(2):131-137
Objective To evaluate the clinical value of holographic image navigation in urological laparoscopic and robotic surgery.Methods The data of patients were reviewed retrospectively for whom accepted holographic image navigation laparoscopic and robotic surgery from Jan.2019 to Dec.2019 in Beijing United Family Hospital and other 18 medical centers,including 78 cases of renal tumor,2 cases of bladder cancer,2 cases of adrenal gland tumor,1 cases of renal cyst,1 case of prostate cancer,1 case of sweat gland carcinoma with lymph node metastasis,1 case of pelvic metastasis after radical cystectomy.All the patients underwent operations.In the laparoscopic surgery group,there were 27 cases of partial nephrectomy,1 case of radical prostatectomy,2 cases of radical cystectomy and 2 cases of adrenalectomy.In the da Vinci robotic surgery group of 54 cases,there were 51 cases of partial nephrectomy,1 case of retroperitoneal lymph node dissection,1 case of retroperitoneal bilateral renal cyst deroofing and 1 case of resection of pelvic metastasis.There were 41 partial nephrectomy patients with available clinical data for statistic,with a median age of 53.5 years (range 24-76),including 26 males and 15 females.The median R.E.N.A.L score was 7.8 (range 4-11).Before the operation,the engineers established the holographic image based on the contrast CT images and reports.The surgeon applied the holographic image for preoperative planning.During the operation,the navigation was achieved by real time fusing holographic images with the laparoscopic surgery images in the screen.Results All the procedures had been complete uneventfully.The holographic images helped surgeon in understanding the visual three-dimension structure and relation of vessels supplying tumor or resection tissue,lymph nodes and nerves.By manipulating the holographic images extracorporeally,the fused image guide surgeons about location vessel,lymph node and other important structure and then facilitate the delicate dissection.For the 41 cases with available clinical data including 23 cases of robotic-assisted partial nephrectomy and 18 cases of laparoscopic nephrectomy,the median operation time was 140 (range 50-225) min,the median warm ischemia time was 23 (range 14-60) min,the median blood loss was 80(range 5-1 200) ml.In the robotic surgery group,the median operation time was 140 (range 50-215)min,the median warm i schemia time was 21 (range 17-40)min,the median blood loss was 150(range 30-1 200)ml.In the laparoscopic surgery group,the median operation time was 160(range 80-225)min,the median warm ischemia time was 25 (range 14-60)min,the median blood loss was 50 (range 5-1 200) ml.All the patients had no adjacent organ injury during operation.There were 2 cases with Clavien Ⅱ complications.One required transfusion and the other one suffered hematoma post-operation.However,the tumors were located in the renal hilus for these 2 cases and the R.E.N.A.L scores were both 11.Conclusions Holographic image navigation can help location and recognize important anatomic structures during the surgical procedures..This technique will reduce the tissue injury,decrease the complications and improve the success rate of surgery.