1.siRNA inhibits HPV16 E6 expression and cell proliferation in HNE-1 cell line
Journal of Chongqing Medical University 2007;0(08):-
Objective:To evaluate the inhibitory effect of Small interferirng RNA(siRNA) on human papilloma vinus(HPV)16 E6 oncogenes in nasopharyngeal carcinoma(NPC) cell line HNE-1,and observe the effects of HPV16 E6 silencing on NPC cell growth.Methods:Four siRNA against the HPV16 E6 gene were designed and transfected into HNE-1 cell respectively using RNAi-Mate transfection reagent.MTT was used to investigate the cellular proliferation after the transfection.RT-PCR was used to detect the expression of HPV16 E6 and protein level of HPV16 E6 was measured by Immunohistochemical staining.Results:After the most effective siRNA was transfected into HNE-1cell,the result of MTT indicated that the cellular proliferation was restrained remarkably,and the highest inhibitory rate was 32%.Meanwhile,RT-PCR and immunohistochemical staining showed that there was a significant decrease in HPV16 E6 mRNA and protein level.Conclusion:These results demonstrated that siRNA against HPV16 E6 could effectively downregulate HPV16 E6 expression in HNE-1 cell line,and inhibit the cellular proliferation.RNAi has great potential in the treatment of nasopharyngeal carcinoma as a new gene therapy.
2. Immunomodulation of bone marrow mesenchymal stem cells
Academic Journal of Second Military Medical University 2010;30(6):717-721
Bone marrow mesenchymal stem cells (BMSCs) are multipotent nonhematopoietic progenitor cells capable of differentiating into multiple lineages, such as bone, fat, and cartilage. BMSCs preferentially home to the damaged tissue and are beneficial to tissue repair. In vitro studies have shown that they do not induce immune response and can inhibit immune cells involved in alloantigen recognition and elimination. In animal models, BMSCs have been shown to induce peripheral tolerance and migrate to injured tissues, where they can inhibit the release of pro-inflammatory cytokines and promote the survival of damaged cells. The unique properties of MSCs suggest a role in cell therapy and treatment of immunomediated diseases.
3.Competing endogenous RNA and tumor pathological mechanism
Shu ZHOU ; Na SHEN ; Fanjun CHENG
Journal of International Oncology 2017;44(2):115-117
Competing endogenous RNA (ceRNA) is a class of RNA which includes mRNA,pseudogenes,long non-coding RNA (lncRNA),circular RNA (circRNA).ceRNA weakens its inhibitory effect on mRNA translation through competitive binding with shared microRNA (miRNA).Many studies have confirmed that the disorder of ceRNA is closely related to the occur-ence of breast cancer,gastric cancer,lymphoma and other tumors.With the improvement of researches,ceRNA may be used as a tumor marker of clinical diagnosis and therapeutic target.
4.Value of multi-parametric MRI in the diagnosis of prostatic abscess
Journal of Practical Radiology 2014;(12):2016-2019
Objective To elucidate the MRI appearance of prostatic abscess,the DWI and enhanced MRI features.Methods 12 cases of prostatic abscesses were retrospectively analyzed,the clinical symptom mainly manifested as lower urinary tract symptoms and fever.All of the patients were given routine MR examination including DWI sequence,6 patients received further enhanced MR examination.Results In the 12 cases,there were 4 cases behaved as single type,8 cases as multifocal type.The abscess showed iso-or slightly hypo-signal intensity on T1 WI,hyper-signal intensity on T2 WI,markedly high signal intensity on DWI and correspond-ing markedly low signal intensity on ADC.Complete abscess walls showed iso-or slightly hyper signal on T1 WI,hypo-signal inten-sity on T2 WI.The mature abscess walls were thin and smooth,which showed homogeneously ring enhanced in 4 cases.The imma-ture abscess walls showed uneven thickness and moderately enhanced in 2 cases.Septum in the abscess could be found in 4 cases, which showed similar enhancement to the abscess walls,while the abscess cavity showed non-enhanced.Abscesses involved the sur-rounding structures in 2 cases,the involved area showed obvious hyper-signal on T2 WI fat-suppression sequence.Conclusion DWI is the best sequence in the diagnosis of prostatic abscess,the markedly high signal intensity on DWI is the characteristic sign.The enhanced MRI showed the walls and septa clearly,the extent and involvement of adjacent structures.The multi-parametric MRI is a prominent procedure in the diagnosis of prostatic abcess.
5.Changes of bone marrow microenvironment of myelodysplastic syndromes
Na SHEN ; Qing LI ; Fanjun CHENG
Journal of International Oncology 2016;43(9):715-717
Bone marrow microenvironment is a complex network consisting of hematopoietic stem/pro-genitor cells (HSPCs),non-hematopoietic cells,extracellular matrix and various cytokines.Its components interact to support normal hematopoiesis.Emerging evidence indicates that the dysfunction of mesenchymal stem cells,myeloid-derived suppressor cells,cytokines and the epigenetic alterations of HSPCs in the bone marrow microenvironment could influence normal hematopoiesis.Abnormal hematopoiesis contributes to the occurrence of hematological malignancies,such as myelodysplastic syndromes (MDS).Animal models have confirmed that bone marrow microenvironment plays an important role in the original generation and maintenance of malignant diseases of hematopoietic system.
6.Pathogenesis and therapy in sepsis-induced cardiomyopathy
Weilan NA ; Huili SHEN ; Dong QU
International Journal of Pediatrics 2021;48(2):95-98
Sepsis-induced cardiomyopathy(SICM)is a reversible cardiac insufficiency in the early stage of sepsis, and mainly manifests as left ventricular dilation, decreased ejection fraction, and recovery within 7~10 days.Although it is reversible, the incidence and mortality in sepsis are high.The specific mechanism is still unclear.Inflammatory reaction, mitochondrial dysfunction, apoptosis and other pathophysiological processes play an important role.Its process is complex and involves the interaction between organism and pathogen.The management of SICM is still based on the etiologic treatment of septic shock guided by hemodynamic monitoring and tissue perfusion, with cardio-protective therapy and specific measures.This review summarizes the literatures on the mechanisms and treatments of SICM.
7.Effect ofα1-antitrypsin Z variant overexpression on autophagy regulation
Na ZHU ; Lijie FENG ; Haiping WANG ; Yujun SHEN ; Yuxian SHEN
Chinese Pharmacological Bulletin 2014;(7):921-925
Aim To investigate the effect of α1-anti-trypsin Z variant (ATZ)overexpression on cell autoph-agy.Methods HEK 293T cells were transfected with pcDNA3.1 zeo+/ATM or pcDNA3.1 zeo+/ATZ,e-qual amount of empty vector was used as control.Cells were treated with NH4Cl for 4 hours and processed for detecting ATZ,LC3 and p62 by immunoblot.Mean-while ,expression and intracellular localization of ATZ, LC3 in 293 T cells were observed with double labeled immunofluorescence.The mRNA levels of autophagy-related genes were measured by real-time PCR.Immu-nohistochemistry was used to observe the morphology of ATZ-positive cells.Results Compared with the control,higher LC3Ⅱ levels and LC3 puncta were observed in ATZ transfected cells.Meanwhile,the levelsof p62 were decreased in ATZ transfected cells,andreversed by NH4 Cl (25 mmol·L -1 )treatment.Overexpression of ATZ increased the mRNA levels of Atg5and Atg12,but had no obvious influence on Beclin1.ATZoverexpressing cells presented abnormal morphologies.The nuclei became reduced,condensed,and even disappeared in ATZpositive cells.Conclusion ATZ overexpression increases autophagy activity whichmay be related to increasing Atg5 and Atg12 levels.
8.Determination of serum acetaminophen based on the diazo reaction and its application in the evaluation of gastric emptying.
Cai-na LI ; Su-juan SUN ; Zhu-fang SHEN
Acta Pharmaceutica Sinica 2015;50(5):560-564
This study aims to establish a method to determine the serum acetaminophen concentration based on diazo reaction, and apply it in the gastric emptying evaluation. Theoretically, acetaminophen could take hydrolysis reaction in hydrochloric acid solution to produce p-aminophenol, which could then take diazo reaction resulting in a product with special absorption peak at 312 nm. Then the serum acetaminophen concentration and recovery rate were calculated according to the standard curve drawn with absorbance at 312 nm. ICR mice were given a dose of acetaminophen (500 mg x kg(-1)) by gavage and the serum acetaminophen was dynamically measured through the diazo reaction. Besides, ICR mice were subcutaneously injected with the long-acting GLP-1 analog GW002 before the gavage of acetaminophen, and serum acetaminophen concentration was measured as above to study how GW002 could influence the gastric emptying. The data showed acetaminophen ranging from 0 to 160 μg x mL(-1) could take diazo reaction with excellent linear relationship, and the regression equation was y = 0.0181 x +0.0104, R2 = 0.9997. The serum acetaminophen was also measured with good linear relationship (y = 0.0045 x + 0.0462, R = 0.9982) and the recovery rate was 97.4%-116.7%. The serum concentration of acetaminophen reached peak at about 0.5 h after gavage, and then gradually decreased. GW002 could significantly lower the serum acetaminophen concentration and make the area under the concentration-time curve (AUC) decrease by 28.4%. In conclusion, a method for the determination of serum acetaminophen based on the diazo reaction was established with good accuracy and could be used in the evaluation of gastric emptying.
Acetaminophen
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blood
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pharmacokinetics
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Aminophenols
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Animals
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Gastric Emptying
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Mice
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Mice, Inbred ICR
9.Clinical analysis of early death in multiple myeloma
Na AN ; Xin LI ; Man SHEN ; Zhongxia HUANG ; Shilun CHEN
Chinese Journal of Clinical Oncology 2016;43(23):1040-1044
Objective:This study investigated the clinical characteristics of multiple myeloma with early death in the era of novel drugs. Methods:Medical records from 188 patients diagnosed from January 2009 to December 2015 were retrospectively reviewed, showing that early death occurred in 19 patients. Early death was defined as death by any cause within the first year after diagnosis. Results:(1) Early mortality was 10.1%, and the median age was 67 years old (range:40-84 years). Eight cases presented IgG type, and 11 cases were non-IgG type. All 19 patients were diagnosed to be at stageⅢin accordance with the Durie–Salmon staging system, and renal insufficiency occurred in 10 patients. In accordance with the International Staging System (ISS), four patients were diagnosed to be at stageⅡ, whereas 15 other patients were at stageⅢ. Extramedullary plasmacytoma (EMP) occurred in six cases, whereas 10 cases pre-sented high-risk patients with cytogenetic abnormalities. Elevated lactate dehydrogenase (LDH) was found in five cases, amyloidosis was detected in three patients, and secondary plasma cell leukemia was observed in two cases. The median score of performance sta-tus (KPS) was 70 (range: 20-80). A total of 16 patients were treated with bortezomib, and 3 patients were treated with CADT. (2) Among the 13 patients who were evaluated, the overall response rate was 46.2%(6/13), and the complete response (CR) and near-CR rate was 7.7%(1/13). (3) The median overall survival was 3 (1-11.5) months, although the two patients with secondary plasma cell leu-kemia survived for less than 2 months. (4) Eight patients died of disease progression (42.1%), eight patients died of severe infections (42.1%), and three patients died of thrombotic events. Conclusion:The important causes of early death include the following:high-risk cytogenetics, elevated LDH, EMP, amyloidosis, advanced age, poor performance status, and serious complications during treat-ment. In the era of novel drugs, we should improve early diagnosis rates and explore individualized treatment for high-risk multiple my-eloma for the benefit of a wide range of patients.
10.The relationships of pulmonary arterial pressure with serum S100B protein, cytokines and procalcitonin in patients with acute exacerbation of chronic obstructive pulmonary disease
Guoping WANG ; Lining SHEN ; Wanping WANG ; Shuli BAI ; Na CUI
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2015;(1):51-54
Objective To investigate the relationships of pulmonary arterial pressure (PAP) with serum protein S100B, cytokines and plasma procalcitonin (PCT) in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). Methods A prospective controlled study was conducted, 160 subjects admitted in the Critical Care Medicine and Respiratory Disease Departments in the Affiliated Hospital of Shanxi Medical University/Changzhi Municipal People's Hospital from January 2012 to August 2013 were enrolled in the study, including 80 patients with AECOPD (AECOPD group) and 80 COPD under stable condition (SCOPD group). Meanwhile 100 healthy people having passed physical examinations were chosen as healthy control group. The levels of blood routine and plasma PCT were examined, PAP was evaluated by modified Simpson, sequation with echocardiography, serum S100B was measured by radioimmunoassay, and enzyme linked immunosorbent assay (ELISA) was used to measure interleukins (IL-18, IL-1β) and tumor necrosis factor-α(TNF-α). The linear correlation analysis was carried out for the various indicators. Results The gender and age in different groups were matched. Compared with the healthy control group, the levels of white blood cell count (WBC), ratio of neutrophil granulocyte (PMN), PAP, PCT and S100B, IL-18, IL-1β, and TNF-αwere significantly higher in SCOPD and AECOPD groups [WBC (×109/L):0.84±0.22, 1.94±0.64 vs. 0.73±0.12, PMN: 0.70±0.09, 0.85±0.08 vs. 0.54±0.05, PAP (mmHg, 1 mmHg = 0.133 kPa): 39±5, 47±8 vs. 24±5, PCT (μg/L): 0.41±0.08, 6.35±2.14 vs. 0.11±0.01, S100B (μg/L): 0.081±0.017, 0.101±0.028 vs. 0.041±0.011, IL-18 (ng/L): 162±19, 181±27 vs. 112±19, IL-1β(ng/L): 55±12, 75±14 vs. 34±10, TNF-α(ng/L):67±17, 89±18 vs. 35±17, all P<0.05], and the increase in level of indexes was more significant in AECOPD group than that in the SCOPD group (all P < 0.01). Serum S100B was significantly positively correlated with PCT, IL-18, PMN and PAP (r value was 0.36, 0.41, 0.39, 0.35, all P<0.05), and plasma PCT was also significantly positively correlated with PMN and PAP (r value was 0.41, 0.37, both P<0.05). Conclusion The level of serum S100B might have positive obvious correlation to the changes of plasma PCT, cytokines and PAP.